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Clinacanthus nutans (Burm. f.) Lindau, a traditional herb renowned for its anti-tumor, antioxidant, and anti-inflammatory properties, has garnered considerable attention. Although its hepatoprotective effects have been described, there is still limited knowledge of its treatment of acute liver injury (ALI), and its mechanisms remain unclear. To assess the efficacy of Clinacanthus nutans in ALI and to identify the most effective fractions and their underlying mechanism of action. Bioinformatics was employed to explore the underlying anti-hepatic injury mechanisms and active compounds of Clinacanthus nutans. The binding ability of schaftoside, a potential active ingredient in Clinacanthus nutans , to the core target nuclear factor E2-related factor 2 (Nrf2) was further determined by molecular docking. The role of schaftoside in improving histological abnormalities in the liver was observed by H&E and Masson's staining in an ALI model induced by CCl 4. Serum and liver biochemical parameters were measured using AST, ALT and hydroxyproline kits. An Fe2+ kit, transmission electron microscopy, western blotting, RT-qPCR, and DCFH-DA were used to measure whether schaftoside reduces ferroptosis-induced ALI. Subsequently, specific siRNA knockdown of Nrf2 in AML12 cells was performed to further elucidate the mechanism by which schaftoside attenuates ferroptosis-induced ALI. Bioinformatics analysis and molecular docking showed that schaftoside is the principal compound from Clinacanthus nutans. Schaftoside was shown to diminish oxidative stress levels, attenuate liver fibrosis, and forestall ferroptosis. Deeper investigations revealed that schaftoside amplified Nrf2 expression and triggered the Nrf2/GPX4 pathway, thereby reversing mitochondrial aberrations triggered by lipid peroxidation, GPX4 depletion, and ferroptosis. The lead compound schaftoside counters ferroptosis through the Nrf2/GPX4 axis, providing insights into a novel molecular mechanism for treating ALI, thereby presenting an innovative therapeutic strategy for ferroptosis-induced ALI. [Display omitted] • Network pharmacology studies have demonstrated the hepatoprotective effects of schaftoside in Clinacanthus nutans. • Ferroptosis is responsible for CCl 4 -induced acute liver injury. • Schaftoside alleviates CCl 4 -induced ferroptosis in acute liver injury. • Schaftoside inhibits ferroptosis via the Nrf2-GPX4 pathway to protect against acute liver injury. [ABSTRACT FROM AUTHOR]

The present study used in vitro and in silico techniques, as well as the metabolomics approach to characterise α-glucosidase inhibitors from different fractions of Clinacanthus nutans. C. nutans is a medicinal plant belonging to the Acanthaceae family, and is traditionally used to treat diabetes in Malaysia. n- Hexane, n- hexane: ethyl acetate (1:1, v/v), ethyl acetate, ethyl acetate: methanol (1:1, v/v), and methanol fractions were obtained via partitioning of the 80% methanolic crude extract. The in vitro α-glucosidase inhibitory activity was analyzed using all the fractions collected, followed by profiling of the metabolites using liquid chromatography combined with mass spectrometry. The partial least square (PLS) statistical model was developed using the SIMCA P+14.0 software and the following four inhibitors were obtained: (1) 4,6,8-Megastigmatrien-3-one; (2) N-Isobutyl-2-nonen-6,8-diynamide; (3) 1′,2′-bis(acetyloxy)-3′,4′-didehydro-2′-hydro-β, ψ-carotene; and (4) 22-acetate-3-hydroxy-21-(6-methyl-2,4-octadienoate)-olean-12-en-28-oic acid. The in silico study performed via molecular docking with the crystal structure of yeast isomaltase (PDB code: 3A4A) involved a hydrogen bond and some hydrophobic interactions between the inhibitors and protein. The residues that interacted include ASN259, HID295, LYS156, ARG335, and GLY209 with a hydrogen bond, while TRP15, TYR158, VAL232, HIE280, ALA292, PRO312, LEU313, VAL313, PHE314, ARG315, TYR316, VAL319, and TRP343 with other forms of bonding. fx1 • The antidiabetic potential of various extracts of C. nutans leaf has been evaluated. • Multivariate data analysis identified 4 α-glucosidase inhibiting compounds. • In silico study predicted the protein-inhibitors interaction. [ABSTRACT FROM AUTHOR]

Graphical abstract Abstract The analytical method used in the quality control of Clinacanthus nutans leaves has not been well developed. Therefore, this study aimed to develop a simple analytical method to predict α-glucosidase inhibitory activity of this herb based on its infrared fingerprinting. The dried extracts obtained from maceration using solvents with different polarities were evaluated for the α-glucosidase inhibitory activity and analysed through infrared spectroscopy. Multivariate data analysis was performed by correlating the bioactivity and infrared spectrum of each extract using partial least square method. The loading plot from multivariate data analysis revealed that C H and C O infrared signals from terpenoids in the extract were positively correlated with the α-glucosidase inhibitory activity. The developed partial least square model was validated through a testing on the external samples. The result concludes that the developed model is valid and capable of predicting α-glucosidase inhibitory activity of the external samples. [ABSTRACT FROM AUTHOR]

The present study sought to identify the key biomarkers and pathways involved in the induction of allergic sensitization to ovalbumin and to elucidate the potential anti-anaphylaxis property of Clinacanthus nutans (Burm. f.) Lindau water leaf extract, a Southeast Asia herb in an in vivo ovalbumin-induced active systemic anaphylaxis model evaluated by 1 H-NMR metabolomics. The results revealed that carbohydrate metabolism (glucose, myo-inositol, galactarate) and lipid metabolism (glycerol, choline, sn-glycero-3-phosphocholine) are the key requisites for the induction of anaphylaxis reaction. Sensitized rats treated with 2000 mg/kg bw C. nutans extract before ovalbumin challenge showed a positive correlation with the normal group and was negatively related to the induced group. Further 1 H-NMR analysis in complement with Kyoto Encyclopedia of Genes and Genomes (KEGG) reveals the protective effect of C. nutans extract against ovalbumin-induced anaphylaxis through the down-regulation of lipid metabolism (choline, sn-glycero-3-phosphocholine), carbohydrate and signal transduction system (glucose, myo-inositol, galactarate) and up-regulation of citrate cycle intermediates (citrate, 2-oxoglutarate, succinate), propanoate metabolism (1,2-propanediol), amino acid metabolism (betaine, N,N -dimethylglycine, methylguanidine, valine) and nucleotide metabolism (malonate, allantoin). In summary, this study reports for the first time, C. nutans water extract is a potential anti-anaphylactic agent and 1 H-NMR metabolomics is a great alternative analytical tool to explicate the mechanism of action of anaphylaxis. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Clinacanthus nutans (Burm. f.) Lindau, a widely used medicinal plant, is extensively grown in tropical Asia and Southeast Asian countries. C. nutans , with its broad spectrum of pharmacological activities, has been traditionally used to treat cancer, inflammatory disorders, diabetes, insect bites, and skin problems, consumed as a vegetable, mixed with fresh juices, in concoctions, and as a whole plant. The present review analyzes the advances in the ethnopharmacology, phytochemistry, pharmacology, and toxicology of C. nutans . In addition, the needs and perspectives for future investigation of this plant are addressed. Aim of the review This review aims to provide a comprehensive report on the ethnomedicinal use, phytochemistry, pharmacological activities, molecular mechanisms, and nutritional values of C. nutans . The present review will open new avenues for further in-depth pharmacological studies of C. nutans for it to be developed as a potential nutraceutical and to improve the available products in the market. Material and methods All the available information on C. nutans was collected using the key words “ Clinacanthus nutans ” and/or “ethnomedicine” and/or “phytochemicals” and/or “anticancer” and/or “anti-inflammatory” and/or “antiviral” through an electronic search of the following databases: PubMed, Web of Science, EMBASE, Cochrane Library, Clinical Trials.org, SciFinder Scholar, Scopus, and Google Scholar. In addition, unpublished materials, Ph.D. and M.Sc. dissertations, conference papers, and ethnobotanical textbooks were used. The Plant List ( www.theplantlist.org ) and International Plant Name Index databases were used to validate the scientific name of the plant. Results The literature supported the ethnomedicinal uses of C. nutans as recorded in Thailand, Indonesia, and Malaysia for various purposes. Bioactivities experimentally proven for C. nutans include cytotoxic, anticancer, antiviral, anti-inflammatory, immunomodulatory, antidiabetic, antioxidant, antihyperlipidemic, antimicrobial, and chemotherapeutic (in aquaculture) activities. Most of these activities have so far only been investigated in chemical, cell-based, and animal assays. Various groups of phytochemicals including five sulfur-containing glycosides, eight chlorophyll derivatives, nine cerebrosides, and a monoacylmonogalactosyl glycerol are present in C. nutans . The presence of two glycerolipids, four sulfur-containing compounds , six known flavones, a flavanol, four flavonols, two phytosterols, one polypeptide, and various phenolics and fatty acids largely influences its diverse bioactivities. Numerous reports justify the ethnomedicinal use of C. nutans as an antiviral agent in treating herpes simplex virus and varicella-zoster virus infections and as part of a traditional anticancer anti-inflammatory concoction agent for various inflammatory diseases. C. nutans tea was reported to have a good percentage of carbohydrate, crude protein, minerals, essential amino acids, nonessential amino acids, and essential fatty acids. Acute, subacute, and subchronic toxicity studies demonstrated that oral administration of ethanol and methanol extracts of C. nutans to male Swiss albino mice and male Sprague–Dawley (SD) rats, respectively, did not lead to any toxicity or adverse effects on the animal behavior and organs when used in amounts as high as 2 g/kg. Conclusion The collected literatures demonstrated that, as an important traditional medicine, C. nutans is a promising ethnomedicinal plant with various extracts and bioactive compounds exhibiting multifarious bioactivities. However, it is important for future studies to conduct further in vitro and in vivo bioactivity evaluations systematically, following the standard pharmacology guidelines. It is crucial to elucidate in-depth molecular mechanisms, structure-activity relationships, and potential synergistic and antagonistic effects of multi-component extracts and bioactive constituents derived from C. nutans . Further studies should also focus on comprehensive toxicity that includes long-term effects and adverse effects on target organs of C. nutans and bioactive compounds in correlation with the specific pharmacological effects. [ABSTRACT FROM AUTHOR]

Clinacanthus nutans (Burm. f.) Lindau is a traditional medicinal plant belonging to the Acanthaceae family. Its therapeutic potentials have been increasingly documented particularly the antiviral activity against Herpes Simplex Virus (HSV), anti-cancer, anti-oxidant, anti-inflammatory and immunomodulatory activities. However, majority of these studies used crude or fractionated extracts and not much is known about individual compounds from these extracts and their biological activities. In the present study, we have isolated four compounds (CN1, CN2, CN3 and CN4) from the hexane fractions of C . nutans leaves. Using NMR spectroscopic analysis, these compounds were identified to be shaftoside (CN1), stigmasterol (CN2), β-sitosterol (CN3) and a triterpenoid lupeol (CN4). To determine the immunosuppressive potential of these compounds, their effects on mitogens induced T and B lymphocyte proliferation and the secretion of helper T cell cytokines were examined. Among the four compounds, stigmasterol (CN2) and β-sitosterol (CN3) were shown to readily inhibit T cell proliferation mediated by Concanavalin A (ConA). However, only β-sitosterol (CN3) and not stigmasterol (CN2) blocks the secretion of T helper 2 (Th2) cytokines (IL-4 and IL-10). Both compounds have no effect on the secretion of Th1 cytokines (IL-2 and IFN-γ), suggesting that β-sitosterol treatment selectively suppresses Th2 activity and promotes a Th1 bias. CN3 was also found to significantly reduce the proliferation of both T helper cells (CD4 + CD25 + ) and cytotoxic T cells (CD8 + CD25 + ) following T cell activation induced by ConA. These results suggested that phytosterols isolated from C . nutans possess immunomodulatory effects with potential development as immunotherapeutics. [ABSTRACT FROM AUTHOR]

Allergy is mediated by the crosslinking of immunoglobulins (Ig) -E or -G to their respective receptors, which degranulates mast cells, macrophages, basophils, or neutrophils, releasing allergy-causing mediators. The removal of these mediators such as histamine, platelet-activating factor (PAF) and interleukins (ILs) released by effector cells will alleviate allergy. Clinacanthus nutans (C. nutans), an herbal plant in Southeast Asia, is used traditionally to treat skin rash, an allergic symptom. Previously, we have reported that C. nutans aqueous leaves extract (CNAE) was able to suppress the release of β-hexosaminidase and histamine but not interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-α) in the IgE-induced mast cell degranulation model at 5 mg/mL and above. We also found that CNAE could protect rats against ovalbumin-challenged active systemic anaphylaxis (OVA-ASA) through the downregulation and upregulation of certain metabolites using proton nuclear magnetic resonance (1H-NMR) metabolomics approach. As allergy could be mediated by both IgE and IgG, we further evaluated the anti-allergy potential of CNAE in both in vitro model of IgG-induced macrophage activation and in vivo anaphylaxis models to further dissect the mechanism of action underlying the anti-allergic properties of CNAE. The anti-allergy potential of CNAE was evaluated in in vivo anaphylaxis models of ovalbumin-challenged active systemic anaphylaxis (OVA-ASA) and IgE-challenged passive systemic anaphylaxis (PSA) using Sprague Dawley rats as well as IgG-challenged passive systemic anaphylaxis (IgG-PSA) using C57BL/6 mice. Meanwhile, in vitro model of IgG-induced macrophage activation model was performed using IC-21 macrophages. The release of soluble mediators from both IgE and IgG-mediated pathways were measured using enzyme-linked immunosorbent assay (ELISA). The signaling molecules targeted by CNAE were identified by performing Western blot. IgG, platelet-activating factor (PAF) and IL-6 was suppressed by CNAE in OVA-ASA, but not IgE. In addition, CNAE significantly suppressed PAF and IL-6 in IgG-PSA but did not suppress histamine, IL-4 and leukotrienes C 4 (LTC 4) in IgE-PSA. CNAE also inhibited IL-6 and TNF-α by inhibiting the phosphorylation of ERK1/2 in the IgG-induced macrophage activation model. Overall, our findings supported that CNAE exerts its anti-allergic properties by suppressing the IgG pathway and its mediators by inhibiting ERK1/2 phosphorylation, thus providing scientific evidence supporting its traditional use in managing allergy. [Display omitted] • Allergy can be mediated via the classical IgE and alternative IgG pathways. • Clinacanthus nutans , found in Southeast Asia was used traditionally to treat skin rash, a symptom of allergy. • Clinacanthus nutans aqueous leaves extract suppressed PAF, IL-6, TNF-α of IgG-mediated allergy by suppressing phosphoERK1/2. [ABSTRACT FROM AUTHOR]

A novel polysaccharide–peptide complex CNP-1-2 with molecular weight of 9.17 × 10 4 Da was obtained from Clinacanthus nutans Lindau leaves by hot water extraction, ethanol precipitation, and purification with Superdex 200 and DEAE-Sepharose Fast Flow column chromatography. CNP-1-2 exhibited the highest growth inhibitory effect on human gastric cancer cells SGC-7901 with inhibition ratio of 92.34% and stimulated activation of macrophages with NO secretion level of 47.53 μmol/L among the polysaccharide fractions. CNP-1-2 comprised approximately 87.25% carbohydrate and 9.37% protein. Monosaccharide analysis suggested that CNP-1-2 was composed of l -rhamnose, l -arabinose, d -mannose, d -glucose and d -galactose with a molar ratio of 1.30:1.00:2.56:4.95:5.09. Methylation analysis, FT-IR, and 1 H NMR spectroscopy analysis revealed that CNP-1-2 might have a backbone consisting of 1,4-linked Glcp, 1,3-linked Glcp, 1,3-linked Manp, 1,4-linked Galp, 1,2,6-linked Galp and 1,2,6-linked Galp. Its side chain might be composed of 1-linked Araf, 1,6-linked Galp and 1-linked Rhap residues. AFM (atomic force micrograph) analysis revealed that CNP-1-2 had the molecular aggregation along with branched and entangled structure. [ABSTRACT FROM AUTHOR]

Clinacanthus nutans is a medicinal plant from the Acanthaceae family, and it is well-known as 'Belalai gajah' and Sabah Snake Grass in Malaysia. C. nutans extracts have been reported to exhibit various bioactivities but reports about its immunomodulatory role are limited. Macrophage activation is lacking in certain macrophage-related immunodeficiency disorders, but it occurs constitutively in certain inflammatory diseases. Thus, the authors investigated the potential role of C. nutans extracts in regulating the macrophage cellular response in J774.2 mouse macrophages. J774.2 macrophages were treated with different concentrations (0–1000 µg/mL) of C. nutans ethanol, ethanol-aqueous, and aqueous extracts for 24, 48, and 72 h and the cytotoxicity effect was assayed using PrestoBlue assay. Then, the macrophages were incubated with 32 µg/mL of ethanol, 500 µg/mL of ethanol-aqueous, and 32 µg/mL of aqueous extracts of C. nutans for 48 h. Cytokine secretion, phagocytic activity, and expression of M1 and M2 activation markers were assessed using a multiplexed cytokine bead-based assay, fluorescence microscopy, and flow cytometry. All extracts did not show any significant cytotoxic effect towards J774.2 macrophages within the extract concentration range tested (cell viability >50%). The ethanol-aqueous extract showed the greatest potential for downregulating lipopolysaccharide-induced pro-inflammatory cytokine (IL-1β, IL-6, and IL-12p40) production and CD86 (M1 activation marker) expression level and for stimulating the phagocytic function of J774.2 macrophages. • No significant cytotoxicity of all three C. nutans extracts studied towards J774.2 macrophages. • All extracts downregulated LPS-induced inflammatory cytokine secretion by macrophages. • All extracts did not disrupt phagocytic function of the macrophages. • 50% ethanol-aqueous extract showed strongest anti-inflammatory effects on macrophages. • 50% ethanol-aqueous extract boosted the phagocytic activity of the macrophages. [ABSTRACT FROM AUTHOR]

Summary: Objective: To examine the efficacy of Clinacanthus nutans extracts in treatment of Herpes genitalis and Herpes zoster from randomised clinical trials (RCTs). Methods: Bibliographic databases, including MEDLINE, EMBASE, CINAHL, Cochrane CENTRAL, AMED, WHO trial registry, http://www.clinicaltrial.gov, Thai Index Medicus, and Index Medicus Siriraj library, were searched from their inception dates to February 2010 without language restrictions. Methodological quality of included trials was assessed using Jadad''s quality scale and Cochrane''s risk of bias. Results: Four RCTs (n =286) met our inclusion criteria which include two studies on H. genitalis and the other two on H. zoster; in these studies, a total of 151 patients were assigned to the C. nutans group of H. genitalis trials, a pooled relative risk of C. nutans preparations against placebo for a 3 day-full crusting was 6.62 (95% C.I. 3.83–11.47) and of a 7-day complete healing was 3.77 (95% C.I. 2.46–5.78). In H. zoster, the relative risk for a 3 day-full crusting was 3.21 (IQR 0.97–10.58). Conclusions: This meta-analysis and systematic review suggests some beneficial effects of C. nutans preparations on treatments of H. genitalis and H. zoster. However, more robustly designed trials are needed to substantiate the benefit of these plants, specifically on their active purified compounds, and their potencies and benefits on treatment outcome of H. genitalis and H. zoster. [Copyright &y& Elsevier]

Based on the discovery of antiviral β-galactosyl diglycerides from Clinacanthus nutans leaves, 19 monoglycosyl diglycerides were synthesized and examined for inhibitory effect on herpes simplex virus types 1 and 2 (HSV-1, HSV-2). A study of the structure–activity relationships of the synthetic monoglycosyl diglycerides indicated that the fatty acyl moieties were critical for inhibitory action with higher activity displayed as the acyl groups became more olefinic in character. The sugar moiety was also important for anti-HSV action; however, the type of sugar (glucose or galactose) did not affect activity. The stereochemistry at C-2 of the glycerol backbone displayed no significant effect on anti-HSV activity. Among the compounds synthesized, 1,2-O-dilinolenoyl-3-O-β-d-glucopyranosyl-sn-glycerol showed the highest inhibitory activity against HSV-1 and HSV-2 with IC50 values of 12.5±0.5 and 18.5±1.5 μg/ml, respectively. [Copyright &y& Elsevier]

Clinacanthus nutans Lindau (Family: Acanthaceae) is a medicinal herb widely distributed in the tropic and subtropic areas of Asia. C. nutans is traditionally consumed as vegetable or herbal tea, as well as a folk medicine for anticancer and antifungal activities. However, to date, chemical constituent responsible for observed health beneficial effects of this medicinal plant is not clear. In the current study, 32 compounds (1 − 32), including three new megastigmanes (1 – 3) were isolated from the aerial parts of C. nutans. Their structures were elucidated on the basis of comprehensive NMR, MS, and CD spectroscopic data analysis, as well as chemical hydrolysis. Among the isolates, cycloartane triterpenoids (9 , 10 , and 12) displayed moderate anti-proliferative effects against HepG2 cell growth with IC 50 values ranging from 9.12 to 19.89 μM. Data obtained from flow cytometry analysis and western blotting assays revealed that compounds 9 and 12 induced apoptosis of HepG2 cells by modulating the expression of proteins associated to mitochondrial-mediated apoptotic pathway. Furthermore, megastigmanes 1 , 2 , 7 , and 8 enhanced the anti- Candida albicans activity of amphotericin B (AmB), supporting the synergistic effects between megastigmanes and AmB. This is the first report of anticancer and antifungal potential of cycloartane triterpenoids and megastigmanes in C. nutans , which shed useful insights on the relationship between C. nutans's chemical constituent and its beneficial effects to health. Findings from this study support further development of this medicinal plant for potential pharmaceutical applications. [Display omitted] • Three new megastigmanes (1 – 3) were isolated and identified from C. nutans. • Cycloartane triterpenoids were reported from C. nutans for the first time. • Cycloartane triterpenoids present in C. nutans could trigger apoptosis in HepG2 cells. • Four megastigmanes exhibited synergistic effects with AmB against Candida albicans. [ABSTRACT FROM AUTHOR]

Clinacanthus nutans has attracted Malaysian public interest due to its high medicinal value in the prevention of cancer. Currently, the specific compound or compounds giving rise to the anticancer potential of C. nutans has not been investigated thoroughly. The extraction was carried out by MeOH at room temperature using the powdered bark of C. nutans , while chromatography was carried out on a silica gel RP-18 column using the crude methanolic extract. Six fractions collected from column chromatography were evaluated by MTT assay against two breast cancer cell lines: MDA-MB-231 and MCF-7. Amongst the fractions, A12 and A17 were shown to exhibit the highest activity. Two sulphur-containing compounds, viz., entadamide C (1) and clinamide D (2), were isolated from these fractions. Molecular docking simulation studies revealed that entadamide C and clinamide D could bind favourably to the caspase-3 binding site with the binding energy of −4.28 kcal/mol and −4.84 kcal/mol, respectively. This study provides empirical evidence for the presence of sulphur-containing compounds in the leaves of C. nutans that displayed anticancer effects which explains its ethnomedicinal application against breast cancer. The docking simulation study showed that both compounds could serve as important templates for future drug design and development. [ABSTRACT FROM AUTHOR]

Clinacanthus nutans or Sabah Snake Grass is a small shrub indigenous to tropical Asia. It has been used in the treatment of skin rashes, insect and snake bites, cancer and diabetes. In this study, Fourier transform infrared spectroscopy was used to discriminate the effects of different extraction methods on the chemical composition of C. nutans freeze dried leaves and stems. The samples were extracted using different methods (soaking and sonication) followed by the measurement of its antiplasmodial activity. Based on the absorption peaks, C. nutans extracts comprise of compounds with hydroxyl, methyl, sulfoxide, sulfone, aromatic and carbonyl functionalities which indicate the presence of carbohydrates, terpenes, sulfurous glycosides and aromatic compounds in the extracts. Multivariate data analysis such as orthogonal partial least square (OPLS) was used to correlate the FTIR spectra of the extracts with antiplasmodial activity. The OPLS model exhibited R2Y and Q2Y values of 0.896 and 0.736, respectively. The RMSEE and RMSE CV value were 22.41 and 32.36. The loading line plot of the OPLS model revealed that OH, C O, C H and C C functional group induced the activity, whereas, C N and S O reduced the bioactivity. These preliminary results demonstrated that FTIR spectroscopy can be used as a simple and rapid technique to discriminate C. nutans extracts obtained from different extraction methods which is useful in the quality control during processing of this plant. [ABSTRACT FROM AUTHOR]

Abstract Aims Clinacanthus nutans (C. nutans) has demonstrated anti-inflammatory activity, however, the active compound generating this activity remains unknown. The aim of this study was to identify the bioactive compound in C. nutans responsible for its anti-inflammatory, in-vitro wound healing, and anti-biofilm activities. Main methods A pure compound was isolated from the chloroform extract (CE) of C. nutans leaves by chromatographic techniques and bioassay-guided fractionation. This compound's structure was determined by spectroscopic analyses (FTIR/NMR/HRES-MS). Biological activities were evaluated using cytotoxicity, nitric oxide (NO), wound scratch, anti-microbial activity, and anti-biofilm assays; and the compound's bactericidal depth into the biofilm was visualized by confocal laser scanning microscopy. Key findings CE and its pure isolated compound, purpurin-18 phytyl ester (P18PE), significantly inhibited lipopolysaccharide (LPS)-induced NO production in RAW 264.7 cells at concentrations of 100 μg/ml and 10–100 μg/ml, respectively. These concentrations significantly induced wound closure by human gingival fibroblasts. CE (100–1000μg/ml) and P18PE (1–500 μg/ml) did not inhibit Streptococcus (S.) mutans growth. However, these concentrations significantly reduced S. mutans biofilm formation below 50% at 250 μg/ml for CE, and 25 μg/ml for P18PE (p<0.05). Significance C. nutans contains a bioactive compound, P18PE, which exhibits anti-inflammatory, in-vitro wound healing, and anti-biofilm activities. [ABSTRACT FROM AUTHOR]