Evaluation of hemostatic, anti-inflammatory, wound healing, skin irritation and allergy, and antimicrobial properties of active fraction from the ethanol extract of Chromolaena odorata (L.) R.M. King & H. Rob.

ETHNOPHARMACOLOGICAL RELEVANCE: Chromolaenaodorata (L.) R.M. King & H. Rob, a perennial herb, has been traditionally utilized as a herbal remedy for treating leech bites, soft tissue wounds, burn wounds, skin infections, and dento-alveolitis in tropical and subtropical regions. AIM OF THE STUDY: The present study was to analyze the active fraction of C. odorata ethanol extract and investigate its hemostatic, anti-inflammatory, wound healing, and antimicrobial properties. Additionally, the safety of the active fraction as an external preparation was assessed through skin irritation and allergy tests. MATERIALS AND METHODS: The leaves and stems of C. odorata were initially extracted with ethanol, followed by purification through AB-8 macroporous adsorption resin column chromatography to yield different fractions. These fractions were then screened for hemostatic activity in mice and rabbits to identify the active fraction. Subsequently, the hemostatic effect of the active fraction was assessed through the bleeding time of the rabbit ear artery in vivo and the coagulant time of rabbit blood in vitro. The anti-inflammatory activity of the active fraction was tested on mice ear edema induced by xylene and rat paw edema induced by carrageenin. Furthermore, the active fraction's promotion effect on wound healing was evaluated using a rat skin injury model, and skin safety tests were conducted on rabbits and guinea pigs. Lastly, antimicrobial activities against two Gram-positive bacteria (G+, Staphylococcus aureus and S. epidermidis) and three Gram-negative bacteria (G-, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa) were determined using the plate dilution method. RESULTS: The ethanol extract of C. odorata leaves and stems was fractionated into 30%, 60%, and 90% ethanol eluate fractions. These fractions demonstrated hemostatic activity, with the 30% ethanol eluate fraction (30% EEF) showing the strongest effect, significantly reducing bleeding time (P < 0.05). A concentration of 1.0 g/mL of the 30% EEF accelerated cutaneous wound healing in rats on the 3rd, 6th, and 9th day post-operation, with the healing effect increasing over time. No irritation or allergy reactions were observed in rabbits and guinea pigs exposed to the 30% EEF. Additionally, the 30% EEF exhibited mild inhibitory effect on mice ear and rat paw edema, as well as antimicrobial activity against tested bacteria, with varying minimal inhibitory concentration (MIC) values. CONCLUSIONS: The 30% EEF demonstrated a clear hemostatic effect on rabbit bleeding time, a slight inhibitory effect on mice ear edema and rat paw edema, significant wound healing activity in rats, and no observed irritation or allergic reactions. Antibacterial activity was observed against certain clinically isolated bacteria, particularly the G- bacteria. This study lays the groundwork for the potential development and application of C. odorata in wound treatment.

Chromolaena odorata layered-nitrile rubber polymer transdermal patch enhanced wound healing in vivo.

The objective is to investigate the healing efficacy of a Chromolaena odorata layered-nitrile rubber transdermal patch on excision wound healing in rats. Wounds were induced in Sprague-Dawley rats and were later treated as follows: wound A, the negative control, received no treatment (NC); wound B, the negative control with an empty nitrile rubber patch (NC-ERP); wound C, treated with a C. odorata layered-nitrile rubber patch (CO-NRP); and wound D, the positive control with Solcoseryl gel with a nitrile rubber patch (PC-SG-NRP). After 1, 3, 6, 10, and 14 days, the rats were sacrificed and analyzed for wound contraction, protein content, hexosamine, and uronic acid levels. Macroscopic observation showed enhanced wound healing in wounds treated with CO-NRP with a wound contraction percentage significantly higher (p<0.05) on days 6 and 10 compared to those treated with NC-ERP. Similarly, protein, hexosamine, and uronic acid contents were also significantly higher (p<0.05) in CO-NRP-treated wounds when compared with wounds treated with NC-ERP. Histological findings showed denser collagen deposition and faster granulation tissue formation in wounds treated with CO-NRP. From the results obtained, it is concluded that the C. odorata layered-nitrile rubber transdermal patch was effective in healing skin wounds.

Unveiling the antibacterial and antifungal potential of biosynthesized silver nanoparticles from Chromolaena odorata leaves.

This research investigates the biogenic synthesis of silver nanoparticles (AgNPs) using the leaf extract of Chromolaena odorata (Asteraceae) and their potential as antibacterial and antifungal agents. Characterization techniques like ultraviolet-visible, Fourier transform infrared (FTIR), Dynamic light scattering and zeta potential (DLS), X-ray diffraction (XRD), transmission electron microscopy (TEM), and field emission scanning electron microscopy and energy-dispersive X-ray spectroscopy (FESEM-EDX) confirmed the formation of spherical (AgNPs). UV-vis spectroscopy reaffirms AgNP formation with a peak at 429 nm. DLS and zeta potential measurements revealed an average size of 30.77 nm and a negative surface charge (- 0.532 mV). Further, XRD analysis established the crystalline structure of the AgNPs. Moreover, the TEM descriptions indicate that the AgNPs are spherical shapes, and their sizes ranged from 9 to 22 nm with an average length of 15.27 nm. The X-ray photoelectron spectroscopy (XPS) analysis validated the formation of metallic silver and elucidated the surface state composition of AgNPs. Biologically, CO-AgNPs showed moderate antibacterial activity but excellent antifungal activity against Candida tropicalis (MCC 1559) and Trichophyton rubrum (MCC 1598). Low MIC values (0.195 and 0.390 mg/mL) respectively, suggest their potential as effective antifungal agents. This suggests potential applications in controlling fungal infections, which are often more challenging to treat than bacterial infections. Molecular docking results validated that bioactive compounds in C. odorata contribute to antifungal activity by interacting with its specific domain. Further research could pave the way for the development of novel and safe antifungal therapies based on biogenic nanoparticles.

Anti-malarial and haematological evaluation of the ethanolic, ethyl acetate and aqueous fractions of Chromolaena odorata.

Malaria is a global health challenge with endemicity in sub-Saharan Africa, where there are multiple drug-resistant strains and limited access to modern health care facilities, especially in rural areas. Studies indicate that African traditional medicine could make a substantial contribution to the reduction of malaria-related deaths and achievement of universal health coverage (UHC), particularly in these regions. Thus, this study evaluated the curative antimalarial effects of Chromolaena odorata leaf extract using mouse model. Forty-five (45) albino mice weighing between 18 and 22 g were grouped into nine groups of 5 animals each. Animals in groups 2-9 were infected with the chloroquine-resistant strain of Plasmodium berghei, while animals in groups 3-9 were subsequently treated with 10 mg/kg chloroquine, a combination of 1.4 mg/kg artemether and 8.75 mg/kg lumefantrine (Coartem), and varying concentrations of the fraction from the aqueous leaf extract of C. odorata at day 3 post-infection. The findings from this study indicate that treatment with 400 mg/kg of the ethanolic fraction of the crude extract resulted in a significant decrease in parasite load (97.6%), which was comparable to the activities of the conventional drugs chloroquine (98.6%) and Coartem (98.8%). The ethyl acetate and ethanolic fractions at 400 mg/kg also ameliorated the significant alterations in the red blood cells, white blood cells, and platelets of the infected animals. The high antimalarial activity displayed by the ethanolic fraction could be due to the presence of quercetin and kaempferol, as detected by high performance liquid chromatography (HPLC) analysis. The findings suggest that the fractions from C. odorata could serve as an alternative source of malaria therapy, particularly in sub-Saharan Africa.

Phytochemistry, Biological, and Toxicity Study on Aqueous and Methanol Extracts of Chromolaena odorata.

The medicinal plant Chromolaena odorata is traditionally used by people living in different communities of Nepal and the globe against diabetes, soft tissue wounds, skin infections, diarrhea, malaria, and several other infectious diseases. The present study focuses on the qualitative and quantitative phytochemical analyses and antioxidant, antidiabetic, antibacterial, and toxicity of the plant for assessing its pharmacological potential. The extracts of flowers, leaves, and stems were prepared using methanol and distilled water as the extracting solvents. Total phenolic content (TPC) and total flavonoid content (TFC) were estimated by using the Folin-Ciocalteu phenol reagent method and the aluminum chloride colorimetric method. Antioxidant and antidiabetic activities were assessed using the DPPH assay and α-glucosidase inhibition assay. A brine shrimp assay was performed to study the toxicity, and the antibacterial activity test was performed by the agar well diffusion method. Phytochemical analysis revealed the presence of phenols, flavonoids, quinones, terpenoids, and coumarins as secondary metabolites. The methanol extract of leaves and flowers displayed the highest phenolic and flavonoid content with 182.26 ± 1.99 mg GAE/g, 128.57 ± 7.62 mg QE/g and 172.65 ± 0.48 mg GAE/g, 121.74 ± 7.06 mg QE/g, respectively. The crude extracts showed the highest DPPH free radical scavenging activity with half maximal inhibitory concentration (IC50) of 32.81 ± 5.26 µg/mL and 41.00 ± 1.10 µg/mL, respectively. The methanol extract of the leaves was found to be effective against bacterial strains such as K. pneumoniae (ZOI = 9.67 ± 0.32 mm), B. subtilis (ZOI = 15.00 ± 0 mm), and E. coli (7.3 ± 0.32 mm). The methanol extract of the flowers showed the most α-glucosidase inhibitory activity (IC50 227.63 ± 11.38 µg/mL), followed by the methanol extract of leaves (IC50 249.50 ± 0.97 µg/mL). The aqueous extract of the flowers showed the toxic effect with LC50 107.31 ± 49.04 µg/mL against the brine shrimp nauplii. In conclusion, C. odorata was found to be a rich source of plant secondary metabolites such as phenolics and flavonoids with potential effects against bacterial infection, diabetes, and oxidative stress in humans. The toxicity study showed that the aqueous extract of flowers possesses pharmacological activities. This study supports the traditional use of the plant against infectious diseases and diabetes and provides some scientific validation.

A Novel Tri-Hydroxy-Methylated Chalcone Isolated from Chromolaena tacotana with Anti-Cancer Potential Targeting Pro-Survival Proteins.

Chromolaena tacotana (Klatt) R. M. King and H. Rob (Ch. tacotana) contains bioactive flavonoids that may have antioxidant and/or anti-cancer properties. This study investigated the potential anti-cancer properties of a newly identified chalcone isolated from the inflorescences of the plant Chromolaena tacotana (Klatt) R. M. King and H. Rob (Ch. tacotana). The chalcone structure was determined using HPLC/MS (QTOF), UV, and NMR spectroscopy. The compound cytotoxicity and selectivity were evaluated on prostate, cervical, and breast cancer cell lines using the MTT assay. Apoptosis and autophagy induction were assessed through flow cytometry by detecting annexin V/7-AAD, active Casp3/7, and LC3B proteins. These results were supported by Western blot analysis. Mitochondrial effects on membrane potential, as well as levels of pro- and anti-apoptotic proteins were analyzed using flow cytometry, fluorescent microscopy, and Western blot analysis specifically on a triple-negative breast cancer (TNBC) cell line. Furthermore, molecular docking (MD) and molecular dynamics (MD) simulations were performed to evaluate the interaction between the compounds and pro-survival proteins. The compound identified as 2',3,4-trihydroxy-4',6'-dimethoxy chalcone inhibited the cancer cell line proliferation and induced apoptosis and autophagy. MDA-MB-231, a TNBC cell line, exhibited the highest sensitivity to the compound with good selectivity. This activity was associated with the regulation of mitochondrial membrane potential, activation of the pro-apoptotic proteins, and reduction of anti-apoptotic proteins, thereby triggering the intrinsic apoptotic pathway. The chalcone consistently interacted with anti-apoptotic proteins, particularly the Bcl-2 protein, throughout the simulation period. However, there was a noticeable conformational shift observed with the negative autophagy regulator mTOR protein. Future studies should focus on the molecular mechanisms underlying the anti-cancer potential of the new chalcone and other flavonoids from Ch. tacotana, particularly against predominant cancer cell types.

Global spatial distribution of Chromolaena odorata habitat under climate change: random forest modeling of one of the 100 worst invasive alien species.

Anthropogenic activities and global climate change increase the risk of Chromolaena odorata invasion and habitat expansion. To predict its global distribution and habitat suitability under climate change, a random forest (RF) model was employed. The RF model, utilizing default parameters, analyzed species presence data and background information. The model revealed that the current spatial distribution of C. odorata covers 7,892,447 km2. Predictions for 2061- 2080 indicate expansion of suitable habitat (42.59 and 46.30%), reduction of suitable habit (12.92 and 12.20%), and preservation of suitable habitat (87.08 and 87.80%) under the SSP (Shared Socio-economic Pathway) 2-4.5 and SSP5-8.5 scenarios, respectively, in comparison to the present distribution. Currently, C. odorata is predominantly found in South America, with limited presence in other continents. However, the data suggest that climate change will elevate the global invasion risk of C. odorata worldwide, particularly in Oceania, Africa, and Australia. Countries such as Gambia, Guinea-Bissau, and Lesotho, which currently have unsuitable habitats, are predicted to have highly suitable habitats with climate change, supporting the idea that global habitat expansion for C. odorata will occur due to climate change. This study indicates that proper management of C. odorata is crucial during the early invasion phase.

Changes in arbuscular mycorrhizal fungal communities, mycorrhizal soil infectivity, and phosphorus availability under Chromolaena odorata (Asteraceae) invasions in a West-African forest-savanna ecotone.

Substantial areas of agricultural lands in Sub-Saharan Africa have been invaded by Chromolaena odorata (Asteraceae), but the consequences for arbuscular mycorrhiza fungi (AMF) remains poorly understood. This study explores changes in diverse AMF community attributes and soil available phosphorus following C. odorata invasion in forest and savanna fragments in Côte d'Ivoire (West Africa). Invaded-forest (COF) and savanna (COS) sites were compared to adjacent natural forest (FOR) and savanna (SAV) fragments, respectively. Physico-chemical variables and AMF spore density parameters were determined for soil samples from 0-20 cm depth. An 18S ribosomal RNA metabarcoding analysis of AMF communities was conducted. In addition, cowpea (Vigna unguiculata) was grown on soils collected from these sites under greenhouse conditions for determination of soil mycorrhizal infectivity. Noticeable changes in the composition of AMF communities in C. odorata relative to nearby forest and savanna non-invaded sites were observed. AMF-specific richness in COS (47 species) was lower than that in SAV (57 species) while it was higher in COF (68 species) than in FOR (63 species). COF and COS differed in AMF specific composition (Dissimilarity index = 50.6%). Chromolaena odorata invasions resulted in increased relative abundances of the genera Claroideoglomus and Glomus in COF, a decreased relative abundance of Paraglomus in COS and decreased relative abundances of Ambispora in both COF and COS. Total and healthy spore densities, cowpea root colonization intensity and soil available P were all higher in invaded sites than in natural ecosystems. Remarkably, although these values were different in FOR and SAV, they turned out to be similar in COF and COS (4.6 and 4.2 total spores g-1 soil, 2.3 and 2.0 healthy spores g-1 soil, and 52.6 and 51.6% root colonization, respectively) suggesting a C. odorata-specific effect. These findings indicate that soil mycorrhizal potential and phosphorus availability have improved following C. odorata invasion.

Anti-Inflammatory Effect of Izalpinin Derived from Chromolaena leivensis: λ-Carrageenan-Induced Paw Edema and In Silico Model.

The flavonoid izalpinin was isolated from the aerial parts of Chromolaena leivensis. Its structural determination was carried out using MS and NMR spectroscopic techniques (1H, 13C). This compound was evaluated for its anti-inflammatory effect in a rat model on λ-carrageenan-induced plantar edema. Paw inflammation was measured at one-hour intervals for seven hours following the administration of λ-carrageenan. Serum creatine kinase (CK) levels were evaluated, obtaining statistically significant results with the treatments at doses of 10 mg/kg (* p < 0.01) and 20 mg/kg (** p < 0.005). The anti-inflammatory effect of the compound was evaluated by using plethysmography, and the results showed significant differences at the three concentrations (10 mg/kg, 20 mg/kg, 40 mg/kg) in the first and third hours after treatment. * p < 0.05; ** p < 0.001; **** p < 0.0001 vs. the negative control group treated with vehicle (DMSO). Lastly, molecular docking analyses reveal that izalpinin has a strong binding affinity with five target proteins involved in the inflammatory process. The analysis using molecular dynamics allowed demonstrating that the ligand-protein complexes present acceptable stability, with RMSD values within the allowed range.

Use of activated Chromolaena odorata biomass for the removal of crystal violet from aqueous solution: kinetic, equilibrium, and thermodynamic study.

In the present study, biomass from the Chromolaena odorata plant's stem was activated using sulfuric acid to adsorb crystal violet (CV) dye. The adsorption operation of CV dye was studied considering the effect of variables like pH, initial dye concentration, time, adsorbent dosage, and temperature. The pseudo-second-order equation best fitted the kinetic study. The thermodynamic parameters such as activation energy (9.56 kJ/mol), change in Gibbs energy (81.43 to 96.7 kJ/mol), enthalpy change (6.89 kJ/mol), and entropy change (-254.4 J/mol K) were calculated. Response surface methodology estimated that at pH (4.902), adsorbent dosage (8.33 g/L), dye concentration (82.30 ppm), and temperature (300.13 K) dye removal of 97.53% is possible. FTIR, SEM, XRD, BJH, and BET confirmed adsorption operation. The adsorbent can be reused for 3 cycles effectively. Langmuir isotherm which best fitted the adsorption operation was used for designing a theoretical single-stage batch adsorber for large-scale operation.

The differentiation of mesenchymal bone marrow stem cells into nerve cells induced by Chromolaena odorata extracts.

Background: Mesenchymal stem cells (MSCs) can differentiate into nerve cells with an induction from chemical compounds in medium culture. Chromolaena odorata contains active compounds, such as alkaloids and flavonoids, that can initiate the transformation of MSCs into nerve cells. The aim of this study was to determine the potential of methanol extracted C. odorata leaf to induce the differentiation of bone marrow MSCs into nerve cells. Methods: A serial concentration of C. odorata leaf extract (0.7-1.0 mg/mL) with two replications was used. The parameters measured were the number of differentiated MSCs into nerve cells (statistically analyzed using ANOVA) and cell confirmation using reverse transcription polymerase chain reaction (RT-PCR). Results: The results showed that the C. odorata extract had a significant effect on the number MSCs differentiating into nerve cells ( p < 0.05) on the doses of 0.8 mg/ml with 22.6%. Molecular assay with RT-PCR confirmed the presence of the nerve cell gene in all of the samples. Conclusions: In conclusion, this study showed the potential application of C. odorata leaf extract in stem cell therapy for patients experiencing neurodegeneration by inducing the differentiation of MSCs into nerve cells.

From wandering weeds to pharmacy: An insight into traditional uses, phytochemicals and pharmacology of genus Chromolaena (Asteraceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Chromolaena species, of the Asteraceae family, are distributed across the tropical and the temperate regions of Africa, the Americas, southern Asia, and Australia. Despite "falling out of favour" among the people because of their "weedy" nature, Chromolaena species have indisputable long medicinal history in the treatment of malaria, nasal congestion, inflammation, eye disorders, asthma, cough, flu, headache, and cold. AIM OF THE REVIEW: The aim of this review is to systematically summarize the current knowledge on ethnopharmacology, phytochemistry, pharmacology, toxicology, and real-time scientific applications of the genus Chromolaena after its re-classification from genus Eupatorium, as well as to proffer integrated approaches in maximizing their therapeutic values despite their "weedy" nature. MATERIALS AND METHODS: First, the current species in the genus were verified by "The Plant List" (http://www.theplantlist.org) and "Royal Botanic Gardens, Kew/Missouri Botanical Garden" (http://mpns.kew.org/mpns-portal/). Second, the relevant information on each of the identified species was gathered from following databases: Google Scholar, Online Wiley library, ScienceDirect, SciFinder, Scopus, PubMed. Scientific literature was searched from inception till August 2021. RESULTS: More than 190 phytochemicals have been isolated and identified from 27 species of the genus, including flavonoids, alkaloids, triterpenoids, diterpenoids, sesquiterpenoids, steroids, fatty acids, and coumarins among others. Pharmacological investigations, both in vitro and in vivo, have shown that the extracts and the compounds have antimicrobial, anticancer, antioxidant, insecticidal, anti-inflammatory, and anti-diabetic activities among others. CONCLUSIONS: Many species of genus have potential therapeutic values, and hence they are more than "wandering" weeds. In addition, there is growing interest in the real-time scientific applications of the genus in the production of pharmacological polyherbal products, and this should serve as a stimulus to strategically develop integrated control approaches for preserving these species, with a view of maximizing their therapeutic values and reducing their cost of eradication.

Effects of Drought Stress on the Growth and Heavy Metal Accumulation by Chromolaena odorata Grown in Hydroponic Media.

This study aimed to investigate the effects of drought stress on cadmium (Cd) and zinc (Zn) accumulation in Chromolaena odorata grown in an artificially contaminated nutrient solution for 15 days. Polyethylene glycol (5% PEG) was used as a drought stressor. The presence of PEG did not affect the chlorophyll content and photochemical efficiency, while drought stress induced by PEG caused a decrease in water content in the plant tissues. The bioaccumulation factor (BAF) of Cd were higher than the BAF of Zn and accumulated mainly in the roots of C. odorata. The highest concentrations (4273.7 mg/kg Cd, 2135.4 mg/kg Zn) were found in the 20 mg/L treatment. The results suggested that Cd and Zn accumulation in C. odorata was not affected by PEG, while a translocation factor (TF) value < 1 was caused by either PEG or contaminants. Based on the hydroponic BAF criterion, the study confirmed that C. odorata was useful for phytoremediation of Cd with low drought stress.

Anti Proliferative and Apoptotic Effect of Soluble Ethyl Acetate Partition from Ethanol Extract of Chromolaena Odorata Linn Leaves Against Hela Cervical Cancer Cell line.

BACKGROUND: Cervical cancer is the leading cause of cancer death in women. Chemotherapy is one of the treatment modalities with many side effects. This study aimed to evaluate the anticancer activity of soluble ethyl acetate partition of Chromolaena odorata (C. odorata) leaves on HeLa cells of cervical cancer. MATERIAL AND METHODS: The cytotoxicity activity of the soluble ethyl acetate extract of the C. odorata leaves was analyzed by MTT colorimetry assay. The apoptosis activity was determined by double staining and flow cytometry techniques. Doubling time assay was used to observed HeLa cells proliferation. RESULTS: The IC50 of soluble ethyl acetate partition of this plant was 82.41± 6.73 µg/ml against HeLa cells. The apoptosis activity showed that HeLa cells underwent morphological changes in dose-dependent manner while the highest number of dead cells was observed after treatment with 500 µg/ml of the partition. Flow cytometry analysis showed that treatment with IC50 and 2x IC50 resulted in death of more than 97% cells (p-value <0.05 in both comparisons). Proliferation of HeLa cells was also inhibited following treatment with ½ IC50, IC50, and 2xIC50 in the first 24 hours (p-value <0.05 in all comparison). CONCLUSION: The findings of this study suggested that the soluble ethyl acetate partition from ethanol extract of C. odorata leaves had cytotoxic and antiproliferative properties against HeLa cells.

Therapeutic potential of Chromolaena odorata phyto-constituents against human pancreatic α-amylase.

Type II Diabetes Mellitus (DM) is caused by insulin resistance in peripheral tissue and impaired insulin secretion through a dysfunction of the pancreatic ß-cell. Acarbose is an anti-DM drug, it is effective but its continuous use may lead to undesirable side effects. Hence, the development of novel drugs from natural source that have both anti-diabetic and anti-oxidant activities, with little or no side effect during long-term use is of great importance. To investigate the anti-DM and anti-oxidant phyto-constituents of Chromoleana odorata, e-pharmacophore model was generated using human pancreatic α-amylase (HPA) standard inhibitor, Acarbose to map important pharmacophoric features of HPA, and used to screen several phyto-constituents of C. odorata to match at least 4 sites of the generated hypothesis. Glide and Induced Fit Docking followed by Prime MM-GBSA calculation, drug-likeness and ADME studies were employed for high fitness (>1.0) compounds retrieved from e-pharmacophore screening process. The drug-likeness properties of the lead compounds, Quercetin and Ombuin were analyzed taking into account the Lipinski's and Veber's rules. Further, machine-learning approach was used to generate QSAR model. The computed model, kpls_desc_19 was used to predict the bioactivity (pIC50) of Quercetin and Ombuin. Phyto-constituents of C. odorata; Quercetin and Ombuin have shown better and promising results when compared to that of the standard, acarbose. Based on the present study, orally delivered Quercetin and Ombuin from C. odorata are relatively better inhibitor of HPA, thus they can be a useful therapeutic candidate in the management/treatment of DM when compared to acarbose.Communicated by Ramaswamy H. Sarma.

Physical Characterisation and Stability Study of Formulated Chromolaena odorata Gel.

BACKGROUND: Formulation of topical products for skin delivery that fulfill good formulation criteria has always been a challenge for pharmaceutical scientists. Despite the challenges, gelbased drug delivery offers some advantages such that it is non-invasive, painless, involves avoidance of the first-pass metabolism, and has satisfactory patient compliance. OBJECTIVES: In this study, C. odorata gel and quercetin gel (bioactive flavonoid compound) were successfully formulated and compared with placebo and conventional wound aid gel. The chromatographic profiling was conducted to screen the presence of phytoconstituents. Subsequently, all formulated gels were evaluated for physical characteristics and stability. METHODS: Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) of C. odorata methanolic leaves extract showed a distinct compound separation at a retention time of 8.4min to 34.8 min at 254nm. All gels were characterised by evaluating their rheological properties, including storage modulus, loss modulus, and plastic viscosity. Besides, texture analysis was performed to measure the firmness, consistency, cohesiveness, and viscosity index of the gels. RESULTS: According to the results, C. odorata gel demonstrated better spreadability as compared to the other gels, which required less work and was found to be favourable for application on the skin. Moreover, C. odorata gel showed no changes in organoleptic properties and proven to be stable after 30 days of accelerated stability study at 40°C ± 2°C with Relative Humidity (RH) of 75% ± 5%. CONCLUSION: C. odorata gel was found to be stable, reflecting the combination of materials used in the formulation, which did not degrade throughout the study. This work suggests the potential of this gel as a vehicle to deliver the active ingredients of C. odorata to the skin, which can be further explored as a topical application for antimicrobial wound management or other skin diseases study.

Phytoprostane and phenolic compounds from Chromolaena palmaris.

The chemical investigation of Chromolaena palmaris (Sch.Bip. ex Baker) R.M. King & H. Rob. expands the phytochemical composition knowledge of Chromolaena genus, since this is the first chemical investigation of this species. Twenty-five compounds were identified, including a phytoprostane, 17 flavonoids, 6 phenolic acids, and a caffeoyl-glucoside derivative obtained by classical chromatography and UHPLC-HRMS/MS analysis. Moreover, anti-Mycobacterium tuberculosis and antiproliferative activities of C. palmaris were evaluated. Dichloromethane fraction showed cytotoxicity towards human cancer cell lines, presenting TGI values on glioma (U251) of 27.8 µg mL-1. Furthermore, compounds 1 and 2 exhibited antimicrobial activity against Mycobacterium tuberculosis with MIC of 62.5 and 15.6 µg mL-1, respectively.

A New Flavanone from Chromolaena tacotana (Klatt) R. M. King and H. Rob, Promotes Apoptosis in Human Breast Cancer Cells by Downregulating Antiapoptotic Proteins.

Chromolaena tacotana is a source of flavonoids with antiproliferative properties in human breast cancer cells, the most common neoplasm diagnosed in patients worldwide. Until now, the mechanisms of cell death related to the antiproliferative activity of its flavonoids have not been elucidated. In this study, a novel flavanone (3',4'-dihydroxy-5,7-dimethoxy-flavanone) was isolated from the plant leaves and identified by nuclear magnetic resonance (NMR) and mass spectrometry (MS). This molecule selectively inhibited cell proliferation of triple-negative human breast cancer cell lines MDA-MB-231 and MCF-7 whit IC50 values of 25.3 µg/mL and 20.8 µg/mL, respectively, determined by MTT assays with a selectivity index greater than 3. Early and late pro-apoptotic characteristics were observed by annexin-V/7-AAD detection, accompanied by a high percentage of the Bcl-2 anti-apoptotic protein inactivated and the activation of effector Caspase-3 and/or 7 in breast cancer cells. It was verified the decreasing of XIAP more than Bcl-2 anti-apoptotic proteins expression, as well as the XIAP/Caspase-7 and Bcl-2/Bax complexes dissociation after flavanone treatment. Docking and molecular modeling analysis between the flavanone and the antiapoptotic protein XIAP suggests that the natural compound inhibits XIAP by binding to the BIR3 domain of XIAP. In this case, we demonstrate that the new flavanone isolated from leaves of Chomolaena tacotana has a promising and selective anti-breast cancer potential that includes the induction of intrinsic apoptosis by downregulation of the anti-apoptotic proteins XIAP and Bcl-2. New studies should deepen these findings to demonstrate its potential as an anticancer agent.

Effects of elevated CO2 concentration and nitrogen addition on foliar phosphorus fractions of Mikania micranatha and Chromolaena odorata under low phosphorus availability.

Invasive plants rapidly spread in habitats with low soil phosphorus (P) availability and have triggered a sharp decline in the diversity of native species. However, no studies have explored how widespread invasive species acclimate to low soil P availability via changing foliar P fractions, especially under elevated atmospheric CO2 concentrations ([CO2 ]) and nitrogen (N) deposition. Here, an open-top chamber experiment was conducted to explore the effect of nutrient addition and elevated [CO2 ] on leaf traits and foliar functional P fractions (i.e., Pi, metabolite P, lipid P, nucleic acid P, and residual P) of two aggressive invasive species (Mikania micranatha and Chromolaena odorata). We found that foliar N/P ratios were more than 20, and P addition significantly increased plant biomass. Both results indicated P-limited plant growth at our studied site. Elevated [CO2 ], N and N + P addition greatly increased plant biomass, photosynthetic rates, and photosynthetic P-use efficiency (PPUE) in invasive species, but PPUE decreased with increasing P addition. Nitrogen addition slightly decreased the concentration of leaf total P, decreased foliar residual P, but increased metabolite P concentrations in invasive species. Similar changes in foliar P fractions were found under N + P addition. Phosphorus addition increased foliar P concentrations, which was strongly correlated with an increase in metabolite P concentrations in invasive species. Elevated [CO2 ] alleviated these effects and increased PPUE. The present results suggest that future elevated [CO2 ] and N deposition allow the invasive species to acclimate to low soil P availability and support their successful invasion by greatly reducing P allocation to non-metabolite foliar P fractions (i.e., nucleic acid P and residual P) to meet their demand of metabolite P for photosynthesis and exhibit a high PPUE.