Pharmacological insights into Ipomoea staphylina : Therapeutic activities and the isolated bioactive metabolic compounds

6 This review comprehensively explores I. staphylina's traditional uses, diverse applications, and pharmacological activities. Extensively used in traditional medicine, this plant addresses a range of ailments, including stomach disorders, respiratory issues, and rheumatism. Research 9 has highlighted its significant antimicrobial, anti-inflammatory, antioxidant, antidiabetic, anthelmintic, and analgesic properties. Notably, its antiulcer activity highlights its potential as a novel antiulcer agent, while hepatoprotective and nephroprotective effects suggest therapeutic 12 applications in liver and kidney disorders. Studies on its anti-diabetic potential show significant reductions in blood glucose levels and positive impacts on biochemical markers. The plant's anti-mutagenic activity against base-pair mutations expands its potential applications. The 15 review also discusses the isolation and pharmacological applications of pure compounds identified through LC-MS and NMR analyses. This review identifies I. staphylina as a promising source of bioactive compounds with therapeutic potential, emphasizing the need for further 18 research to isolate and characterize its active constituents.


INTRODUCTION
Plants have long been relied upon by humanity for their medicinal properties and their diverse applications in various aspects of daily life, including food, clothing, flavors, dyes, and more [1].
Traditional herbal medicine, with its roots dating back centuries, forms the basis of modern healthcare systems.Interestingly, even in developed nations such as China, India, and Japan, traditional medicine continues to be practiced [2].Despite the prevalence of synthetic alternatives in today's digital age, many people remain cautious about the side effects and chemical constituents associated with synthetic medicines.Synthetic drugs, including antibiotics, are known to induce adverse effects such as nausea, vomiting, and photosensitivity [3].Consequently, there is a growing interest in natural medicines and compounds that offer potential benefits with fewer side effects and a more organic impact on the human body [4].
Natural products, which can be readily integrated into our daily diet, are easily absorbed and often preferred.According to the World Health Organization, approximately 21,000 plant species worldwide have been utilized for medicinal purposes [5].Remarkably, over 60% to 75% of cancer and infectious disease drugs have been derived from natural sources [6,7].Plantbased drugs have therefore gained significant traction in contemporary drug design and development [8,9].
One notable herb with significant therapeutic potential is Ipomoea staphylina, which holds a prominent place in traditional medicine [10,11].Traditionally, I. staphylina has been employed to address various disorders [10,11].Its distribution spans regions in India, China, and Sri Lanka, as well as numerous parts of Asia and South Asia [12].Recent research has highlighted the antibacterial, anti-diabetic, anti-inflammatory, anti-mutagenic, analgesic, and antioxidant properties of I. staphylina [3].Abundantly found in wastelands and deciduous forests, I. staphylina is easily accessible.It possesses a rich composition of phytochemical constituents, including alkaloids, flavonoids, phenolics, proteins, carbohydrates, glycosides, and saponins [3,13,14].In local languages, I. staphylina is referred to as "Oonan kodi" or "Onan kodi" in Tamil, "Ugina kodi" or "Unang kodi" in Kannada, and "Sunang kodi" in Irula, while it is known as "Morning clustered glory" in English [9,14].Through this literature review, we aim to provide an in-depth exploration of the ethnobotanical and pharmacological properties of I. staphylina, shedding light on its potential benefits in healthcare and therapeutic applications.The clear image represents the different plant parts of the I. staphylina depicted in Figs. 1 and 2.

Traditional uses
I. staphylina has been used for many health issues like stomach disorders, inflammation, purgation, pain, and also in rheumatism [15].Traditionally Dharmapuri (Tamil Nadu) village folk people used stem raw extract for stomach disorders and respiratory disorders.Gingee Hills villagers used leaf latex for foot crack [7].Tribes of Irulas and Palliyars have used I. staphylina roots as an anti-dote (snake bite) [8].Karandamalai villagers used leave decoction for stomach disorders [16].Chenchus tribes treated leave extract for piles (Andhra Pradesh) [17].

Search strategy
In this review, we systematically compiled data from multiple scientific databases, including Web of Science, Science Direct, Google Scholar, and MEDLINE, focusing on I. staphylina.
Topics covered include plant extraction, isolation, characterization, biological activity, and nanoparticle synthesis.Our analysis integrates findings from both in vitro and in vivo studies, with data collected from the inception of these databases up to March 2024.Publications in languages other than English, as well as conference abstracts and dissertations, were excluded from the review.

Study selection
Publications in languages other than English, along with conference abstracts and dissertations, were excluded from consideration.The initial search yielded 60 studies, from which duplicate entries were removed.The screening process involved evaluating titles, abstracts, and full texts according to predefined inclusion criteria, resulting in the final inclusion of 44 relevant studies in the review.

Antimicrobial activity
In a research study conducted by Narra et al, the antimicrobial potential of the crude latex obtained from the stems of I. staphylina was investigated.The study aimed to evaluate its activity against Staphylococcus aureus and E. coli bacteria, two common pathogens known to cause various infections.Different concentrations of the extract were utilized to determine its efficacy in inhibiting the growth of these bacteria [13].The study demonstrated that the ethanol extract of I. staphylina exhibited significant inhibitory activity against S. aureus and E. coli, indicating its potential as a source of novel antimicrobial agents.To evaluate the safety profile of the extract, an MTT assay was conducted using J774.staphylina stems and highlights its potential as a source of natural compounds with antimicrobial properties [13].Given its excellent antimicrobial activity, formulations such as antimicrobial ointment or cream could be developed from this plant.The percentage of inhibition was determined, and the extracts of I. staphylina exhibited 50% lower inhibitory percentages compared to standard drugs [14].The reported anti-inflammatory properties of I. staphylina, both in vivo and in vitro, highlight its importance in inflammation management in medical situations.Its potent activity against important inflammatory enzymes such as COX and LOX-5 makes it an attractive candidate for formulations targeting inflammatory disorders [13][14][15].Given its shown efficacy and minimal toxicity profile, I.

Anti-inflammatory activity
staphylina is a promising candidate for the development of anti-inflammatory formulations, such of I. staphylina demonstrated remarkable efficacy compared to other extracts evaluated in the study [14].
Furthermore, the antioxidant potential of the ethanolic leaf extract of I. staphylina was assessed using a comprehensive range of assays.These included measurements of superoxide radical scavenging, hydroxyl radical scavenging, DPPH assay, ABTS assay, and metal chelating assay.
The extract was subjected to various concentrations, and the IC50 values were determined employing standard protocols.Furthermore, the study conducted by Shobana D et al., in 2021 investigated the antioxidant activity of the ethanolic leaf extract and whole plant extract of I.
staphylina on streptozotocin-induced rats, yielding excellent results [14,18].The findings from the above-mentioned studies provided valuable insights into the antioxidant properties of I.
staphylina, further enhancing its potential therapeutic applications [14].The biochemical measurements in the rats were performed using the methods outlined in    notably reduced blood glucose levels.Biochemical analysis revealed significant decreases in serum creatinine blood urea, and blood urea nitrogen, levels upon treatment with the extract.

Anti-diabetic activity
Additionally, the ethanolic extract led to reductions in serum total protein and liver glycogen levels in streptozotocin-induced diabetic mice.The ethanol, n-butanol, and ethyl acetate extract fractions exhibited decreases in SGOT, ALP, and SGPT activities in diabetic mice.
Furthermore, the ethanol extract and ethyl acetate fraction demonstrated enhancements in GPx, CAT, and SOD levels [19].Biochemical analysis, as outlined in Table 3.

Serum glucose
Glucometer (Accu-Chek Active, India) Serum total cholesterol, total triglyceride, LDL-c, VLDL-c and HDL-c ethyl acetate leaf extract fractions of I. staphylina against nicotinamide-streptozotocin-induced type 2 diabetes.Wistar rats were employed in this study.The findings indicated a significant reduction in blood glucose levels following treatment with the leaf extracts.Additionally, the extracts led to reductions in biochemical markers such as SGOT, SGPT, and ALP.Histological analysis of the liver, kidney, and pancreas, revealed improvements at the cellular level after 21 days of oral administration of I. staphylina extract and its fractions in nicotinamidestreptozotocin induced type 2 diabetic rats [20].According to the studies mentioned above, various parts of the I. staphylina plant demonstrate remarkable antidiabetic activities effective against both type 1 and type 2 diabetes [18][19][20].Through oral administration, these plant parts effectively reduce blood glucose levels.Consequently, the therapeutic potential of I. staphylina in the management of diabetes is evident, suggesting its utility in the development of diabetic medications.This could include the formulation of oral tablets or syrups tailored to regulate blood sugar levels and alleviate symptoms associated with diabetes.

Anthelmitic activity
In research conducted by Gosh S et al. (2013), the distilled water extract of I. staphylina was evaluated for its anthelmitic ability.The extract was tested against Perionyx excavates earthworm at three different concentrations (25, 50, and 100 mg/ml).A standard drug, Piperazine citrate, was used for comparison.The findings indicated that the I. staphylina plant extract did not exhibit any anthelmintic activity on earthworms, as evidenced by the results obtained [21].

A c c e p e d m a n u s c r i p t
Anti-ulcer activity Banerjee A et al. (2015) conducted a scientific investigation to estimate the anti-ulcer activity of a water and ethanolic (3:7) extract derived from I. staphylina.The study employed dried powdered samples of the entire plant to assess its efficacy.Two well-established experimental models, namely pyloric ligation and ethanol-induced gastric ulcer methods, were employed to simulate ulcer conditions.Oral administration of the I. staphylina plant extract was carried out in rats, and the acid index, a key indicator of ulcer severity, was determined using the standard drug Omeprazole for reference.The findings of the study revealed that the I. staphylina extract exhibited significant anti-ulcer properties, as evidenced by a remarkable reduction in acid pH levels.This indicates the plant's potential as a promising candidate for the development of novel anti-ulcer agents [22].

Hepatoprotectivity and Nephroprotectivity
Bag A.K et al. (2013) conducted a study to assess the hepatoprotective and nephroprotective properties of a hydroalcoholic leaf extract from I. staphylina.To assess the nephroprotective and hepatoprotective activity, Wistar albino rats were induced with gentamicin and CCl4, respectively.The administration of the plant extract was done orally, and no signs of toxicity were observed.
For the evaluation of hepatoprotective activity induced by CCl4, the levels of AST, ALP, ALT, and total bilirubin were measured.The I. staphylina plant extract demonstrated a significant decrease in the levels of AST, ALP, ALT, and total bilirubin, indicating its potential hepatoprotective effects [23].
Furthermore, Jayadevi et al. (2019) reported the hepatoprotective activity of an aqueous leaf extract from I. staphylina.In a study conducted on CCl4-induced Wistar rats, the extract showed a reduction in liver enzymes such as SGOT, SGPT, ALP, and bilirubin.It is important to note that both studies highlight the protective properties of I. staphylina extracts on liver and kidney functions, suggesting their potential therapeutic applications in the treatment of hepatic and renal disorders [24].The administration of the hydroalcoholic extract of I. staphylina at a dose of 200 mg/kg resulted in a significant analgesic effect.The observed activity suggests the potential of the extract as an analgesic agent, comparable to the standard drugs used in the study [25].This research approach provides valuable insights into the analgesic properties of the hydroalcoholic extract of I. staphylina, indicating its potential for further exploration and development as a natural analgesic alternative

Adulticidal activity and Larvicidal activity
In research conducted by Santoshkumar et al. (2011), the inhibitory properties of dried leaf extracts of I. staphylina were investigated.Various solvents such as ethyl acetate, hexane, methanol, and acetone were utilized for the extraction process.The evaluation focused on the inhibition of adults of H. bispinosa, the hematophagous fly H. maculata, and the instar larvae of the malaria vector A. subpictus [26].
The results demonstrated that the leaf extracts of I. staphylina exhibited significant inhibitory activity against these targeted organisms.This suggests the presence of bioactive compounds within the plant that have the potential to act as effective agents against these medically important vectors.
The findings from this scientific investigation provide valuable insights into the potential of I. staphylina as a natural source for developing novel insecticidal agents.Further research and exploration of the specific bioactive constituents responsible for the observed inhibitory effects are warranted for the development of effective vector control strategies.The assessment of anti-mutagenic activity was carried out using the Ames method.The results obtained from the study indicated that I. staphylina exhibits anti-mutagenic activity, and the A c c e p e d m a n u s c r i p t extent of this activity depends on the dosage of the plant extract [27].These findings shed light on the potential of I. staphylina as a source of anti-mutagenic compounds.Further research is warranted to identify and isolate the specific constituents responsible for this activity, and to explore their potential for therapeutic applications in mutagenesis-related disorders.
A c c e p e d m a n u s c r i p t Table 4. Isolated compounds of I. staphylina [14].

Nanoparticle synthesis
Over the last few decades, there has been a rise in the fabrication of therapeutic and diagnostic agents based on nanoparticles for various medical uses, with plants and herbs emerging as promising sources for nanoparticle synthesis [39,40].Various biological sources harbour the potential to yield a wide array of nanoparticles, each characterized by unique properties, shapes, and sizes [41,42].In the past few years, much research has been done in the Nanomaterials field using the I. staphylina plant [39][40][41][42].In 2020, Pugazhendhi synthesized silver nanoparticles utilizing I. staphylina leaf extract, employing green chemistry techniques [43].The synthesized nanoparticles underwent characterization via UV-Vis spectroscopy, FTIR, HRTEM, and XRD.The average particle size of the synthesized silver nanoparticles, determined using Scherrer's formula, was found to be approximately 20 nm, a value corroborated by HRTEM analysis (22 nm) [43].

Fig. 3 .
Fig. 3. Pharmacological applications of I. staphylina 2 cells, a standard model for assessing cell viability and cytotoxicity of test compounds.The assay results offered crucial insights into A c c e p e d m a n u s c r i p t the extract's effects on cell viability, supporting its potential therapeutic application.Overall, the study by Narra et al sheds light on the antimicrobial activity of the crude latex from Ipomoea

Firdous
et al. (2012) investigated the anti-inflammatory properties of the ethanolic leaf extract of I. staphylina.The study evaluated both the ethanolic extract and its ethyl acetate and nbutanol fractions through in vivo and in vitro models.The fractions displayed significant antiinflammatory activity in carrageenan-induced paw edema.Additionally, when administered orally at a dose of 200 mg/kg, the extract and its fractions significantly reduced granuloma formation in the cotton pellet-induced granuloma model (P > 0.001).The study also examined the inhibition of TNF-α in LPS-activated RAW 264.7 cells, where the ethanol, n-butanol, and ethyl acetate fractions showed notable inhibitory effects.Cell viability across different concentrations was further assessed using the MTT assay[15].Narra et al. (2014) focused on the stem ethanol extract of I. staphylina to evaluate its antiinflammatory properties through COX activity assay.The COX assay was performed using a specific chemiluminescent substrate to measure the activity.The latex sample (approximately 20 μl) was added and incubated for 2 hours, followed by the addition of 50 μl of the chemiluminescent substrate to determine the activity.The ethanolic extract demonstrated potent inhibitory activity against COX[13].In a study byReddy et al. (2012), the entire plant of I. staphylina was evaluated for its antiinflammatory activities using a modified version of the techniques.The methanolic and hydroalcoholic extracts of I.staphylina were assessed for their ability to inhibit LOX-5 enzyme.

A c c e
p e d m a n u s c r i p t as gel or tablet formulations.These compositions with significant anti-inflammatory activities have the potential to provide effective treatment while minimizing side effects, addressing a critical demand in inflammatory therapy.Anti-oxidant and free radical scavenging activityReddy et al. (2012) conducted a study to investigate the antioxidant activity of methanolic and hydroalcoholic extracts derived from the entire plant of I. staphylina.The DPPH radical scavenging method was employed to assess the free radical activity.Various concentrations (25, 50, 75, and 100 μg/ml) of the fractions were prepared, and their absorbance at 517 nm was measured to determine the activity.The results revealed that the hydroalcoholic stem extract and methanolic leaf extract exhibited noteworthy antioxidant activity compared to other extracts [14].The superoxide radical scavenging activity of the methanolic leaf extract of I. staphylina was investigated using the Nitro Blue Tetrazolium (NBT) riboflavin photoreduction method, following the methodology established by Mccord and Fridovich.Reddy et al., testified that the hydroalcoholic extract derived from the stem (IC50 = 15.54 μg/ml) and leaf (IC50 = 16.02μg/ml)

A c c e p e d m a n u s c r i p t
In a study conducted byShobana D et al. (2021), the ethanolic leaf extract and whole plant extract of I. staphylina were investigated for their antioxidant, peroxide, and anti-diabetic activities.Male Wistar rats were used, and administration of Streptozotocin and diabetes was induced through intraperitoneal.The assessment of biochemical parameters was performed using established methods.The results demonstrated that I. staphylina extracts pointedly reduced glucose levels in diabetic-induced rats and increased antioxidant levels compared to normal rats.Moreover, the extracts exhibited notable reductions in glutathione peroxidase, hyperoxide, and TBARS levels in diabetic rat tissues.Among the extracts, the ethanolic leaf extract of I. staphylina showed the most promising results in diabetic-induced rats[18].Firdous et al., (2014) conducted a study to investigate the anti-diabetic activity of the ethanolic extract of I. staphylina in Swiss albino mice induced with Streptozotocin.The extract was orally administered, and no acute toxicity or signs of toxicity were observed.The ethanolic extract Standard enzymatic (Span Diagnostic, India) A c c e p e d m a n u s c r i p t SGOT, SGPT, serum ALP, total protein, serum urea, and creatinine Standard enzymatic (span diagnostic, India Glycogen content in the liver Spectrophotometric determination of glycogen with o-toluidine reagent Lipid peroxidase (LPO), Superoxide dismutase (SOD), Catalase (CAT) and glutathione peroxidase (GPx).10% ice-chilled potassium chloride solution In a study by Firdous et al. (2016), the ethanol, n-butanol, and ethyl acetate extract fractions of I. staphylina were assessed for their anti-diabetic property.Alloxan-induced diabetic rats were used in the study, and the extracts were administered orally.The extracts significantly reduced the blood glucose levels.Moreover, Firdous et al. (2016) examined the anti-diabetic effects of the ethanol, n-butanol, and et al., (2014) conducted a study to assess the analgesic activity of the hydroalcoholic extract of I. staphylina.The evaluation was performed using established methods including A c c e p e d m a n u s c r i p t abdominal writhing, formalin-induced paw licking, and the Eddy's hot plate test in Swiss albino mice.As reference standards, Acetylsalicylic acid, Pentazocine, and Diclofenac were utilized.

Banerjee
et al. (2020) performed research to investigate the anti-mutagenic activity of the hydroalcoholic (ethanol 7:3 distilled water) leaf extract of I. staphylina against base-pair mutations induced by 2-Nitrofluorene.Two nonvirulent strains of S. typhi were used to evaluate this activity, namely TA1535 & TA1538.Two different concentrations of the plant extract (250 and 500 µg/ml) were employed along with Cyclophosphamide (50 µg/ml) as a reference.

Fig. 4 .
Fig. 4. Isolation of chemical compound from plant extracts

In a recent study
by Lakshmanan N et al. in 2024, silver and CuO nanoparticles were synthesized using the ethanolic extract of I. staphylina for larvicidal activity[44].Employing environmentally sustainable green chemistry methodologies, the researchers validated the nanoparticle structure and size through XRD analysis, while field-emission scanning electron microscopy (FE-SEM) revealed precise nanostructures.Elemental composition was elucidated via energy-dispersive X-ray (EDX) analysis, and UV-vis spectroscopy provided bandgap energy values (3.15 eV for silver, 1.2 eV for CuO nanoparticles).These nanoparticles displayed

Table 1 .
Biological activities of I.

Table 2 .
Thesereported antioxidant activities suggest that extracts from I. staphylina can be formulated into antioxidant drugs.

Table 2 .
Methods used for biochemical measurements in diabetic-induced rats