Search

Camptothetin (CPT) is a quinoline alkaloid originally isolated from the Chinese tree, Camptotheca acuminata Decne. CPT was found to have anticancerous and antiviral properties. Derivatives of natural CPT, including topothecan and irinotecan are used clinically to treat a variety of cancers. Apart from Camptotheca acuminata Decne, CPT production was also found in the perennial plant Ophiorrhiza mungos. In this study we attempted the immobilization of the tissue culture grown callus of Ophiorrhiza mungos for the continuous production of a higher concentration of CPT. As evident from previous studies about the antiviral effects of CPT, we wanted to bioinformatically analyze the binding potential of CPT towards two important proteins of SARS-CoV-2, protease (Mpro) and RNA dependent RNA polymerase (RdRp). Further docking analysis of the CPT against the exterior spike glycoprotein of SARS-CoV-2 was also done to determine their potential interaction. The immobilized callus of Ophiorrhiza mungos produced CPT at a concentration of 420 µg/l by the end of 12 days of growth. The HPLC analysis was done to determine the purity of the CPT synthesized by the immobilization technique. The bioinformatic analysis revealed a higher binding efficiency of CPT and its derivatives, toptecan and irinotecan against Mpro and RdRp. The docking analysis of CPT against the spike glycoprotein of SARS-CoV-2 showed hydrogen bonding with the amino acids at K466 with a bond distance of 2.56A° and K355 with a bond distance of 2.40A°. This finding was of particular importance that other compounds including hydroxychloroquine sulphate, lopinavir and ivermectin could bind with the spike protein only by weak Vander wall bonds and no hydrogen bond formation was noticed. Our studies hence evaluate the efficiency of CPT against SARS-CoV-2, by potentially blocking the interaction of the spike glycoprotein with the angiotensin-converting enzyme 2 (ACE2) receptor found on host cells.

In the study, endophytic fungi isolated from Ophiorrhiza mungos were screened for camptothecin (CPT) biosynthetic potential by high performance liquid chromatography (HPLC). Among the 16 fungi screened, OmF3, OmF4, and OmF6 were identified to synthesize CPT. Further LC–MS analysis also showed the presence of CPT specific m/z of 349 for the extracts from OmF3, OmF4, and OmF6. However, the fragmentation masses with m/z of 320, 305, 277 and 220 specific to the CPT could be identified only for the OmF3 and OmF4. These CPT producing fungi were further identified as Meyerozyma sp. OmF3 and Talaromyces sp. OmF4. The cultures of these two fungi were then supplemented with nanoparticles and analyzed for the quantitative enhancement of CPT production by LC–MS/MS. From the result, Meyerozyma sp. OmF3 was found to produce 947.3 ± 12.66 μg/L CPT, when supplemented with 1 μg/mL zinc oxide nanoparticles and the same for uninduced parental strain OmF3 was only 1.77 ± 0.13 μg/L. At the same time, Talaromyces sp. OmF4 showed the highest production of 28.97 ± 0.37 μg/L of CPT when cultured with 10 μg/mL silver nanoparticles and the same for uninduced strain was 1.19 ± 0.24 μg/L. The observed quantitative enhancement of fungal CPT production is highly interesting as it is a rapid and cost effective method. The study is remarkable due to the identification of novel fungal sources for CPT production and its enhancement by nanoparticle supplementation. [ABSTRACT FROM AUTHOR]

The gastroprotective effect of luteolin-7-O-glucoside (LUT7G) on different ulcer models using rats including the indomethacin model, the ethanol-induced gastric ulcer model and the Shay ulcer model was examined. The ethanol-induced ulcer group showed significant increases in MPO, NO, iNOS, MMP-2, MMP-9, caspase-3, apoptosis, IL-6, TNF-α, IKK, NF-κB p65, and ICAM-1 and declines in PGE2, arginase, SOD, GSH, mucin, IL-10, eNOS, COX-1, HSP-70, and IκBα levels. However, LUT7G (25 mg/kg) pretreatment significantly reverted the pathophysiological levels of these biomarkers to near normal levels. The gastroprotective activity of LUT7G was abolished by pretreatment with SC560, rofecoxib, and L-NAME, demonstrating the participation of COX and NOS in LUT7G-facilitated gastroprotection against ethanol-induced ulcers. Convincingly, LUT7G (25 mg/kg) provided protective effects in the rat gastric mucosa against ethanol-induced gastric injury at least in part to antisecretory, anti-inflammatory, antioxidative, and antiapoptotic activity, and augmentation of PGE2, mucin and HSP-70 synthesis.

Adventitious root cultures of medicinal plants are a source of secondary metabolites of pharmaceutical importance . Ophiorrhiza mungos (Rubiaceae) contains a bioactive compound, camptothecin (CPT), which possess anticancer activity. Adventitious roots of O. mungos were induced on aseptic leaf explants and established isolated root cultures. Various culture parameters were studied and showed that 1/2th strength MS liquid media with 5.0 % (w/v) sucrose, 0.5 mg L −1 IBA, 0.25 mg L −1 1-Naphthaleneacetic acid (NAA) and 0.1 mg L −1 Gibberellic Acid (GA 3 ) and an inoculum size of 2 g L −1 was ideal for enhanced biomass combined and CPT yield. The concentration of ammonium to nitrate at 20:40 mM favoured root growth and 50:10 mM favoured CPT accumulation. A significant increase in root growth was observed at 1.25 mM phosphate and CPT accumulation was unaffected. Roots were grown in a modified nutrient medium (MS1) according to the results obtained with the individual culture parameters and obtained significantly high biomass and CPT production. CPT was estimated using high performance liquid chromatography (HPLC). The results revealed the effect of major nutrients, growth regulators and inoculum size on growth and CPT production. The study showed that adventitious root culture of O. mungos is a viable alternative culture method for CPT production and results are useful in the scaling up studies using suitable bioreactor systems for the large-scale production of CPT. [ABSTRACT FROM AUTHOR]

The gastroprotective effect of luteolin-7-O-glucoside (LUT7G) on different ulcer models using rats including the indomethacin model, the ethanol-induced gastric ulcer model and the Shay ulcer model was examined. The ethanol-induced ulcer group showed significant increases in MPO, NO, iNOS, MMP-2 , MMP-9 , caspase-3, apoptosis, IL-6 , TNF-α, IKK, NF-κB p65, and ICAM-1 and declines in PGE 2 , arginase, SOD, GSH, mucin, IL-10, eNOS, COX-1, HSP-70, and IκBα levels. However, LUT7G (25 mg/kg) pretreatment significantly reverted the pathophysiological levels of these biomarkers to near normal levels. The gastroprotective activity of LUT7G was abolished by pretreatment with SC560, rofecoxib, and L-NAME, demonstrating the participation of COX and NOS in LUT7G-facilitated gastroprotection against ethanol-induced ulcers. Convincingly, LUT7G (25 mg/kg) provided protective effects in the rat gastric mucosa against ethanol-induced gastric injury at least in part to antisecretory, anti-inflammatory, antioxidative, and antiapoptotic activity, and augmentation of PGE 2 , mucin and HSP-70 synthesis. [ABSTRACT FROM AUTHOR]

An efficient protocol has been established for rapid multiplication of Ophiorrhiza mungos Linn. (Rubiaceae), a potent anticancer plant. Axillary and terminal buds of these in vitro-raised seedlings formed the primary source of explants for direct organogenesis. Explants were inoculated onto MS medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BA) and kinetin (KN). The best morphogenic response was observed on MS media supplemented with 0.25 mg L BA and 0.25 mg L KN, which exhibited the highest regeneration frequency (84%), the maximum number of shoots/explants (63.1 ± 1.35) and shoot length (2.8 ± 1.15) within 4 weeks. Fortification of 1.0 mg L GA enhanced the shoot elongation by 2.33 fold in 91% of shoot cluster cultures within 3 weeks. A high percent frequency of rooting (92.13%) was achieved within 15 days of shoot implantation on ½ strength MS media fortified with 100 mg L activated charcoal. The rooted plantlets were successfully acclimatized with 95% survival rate. Randomly amplified polymorphic DNA (RAPD) analysis confirmed that all the regenerated plants were genetically identical to their mother plant, suggesting an absence of detectable genetic variation in the regenerated plantlets. High performance liquid chromatography (HPLC) was done to further confirm the existence of qualitative and quantitative differences in the major secondary metabolite (camptothecin) between the mother plant and in vitro-propagated plants. The present results evidently showed comparable chemical profiles. Thus, the present protocol can be used for clonal mass propagation of true-to-type elite O. mungos plants. [ABSTRACT FROM AUTHOR]

High-performance liquid chromatography is the prevalent method used to estimate Camptothecin quantitatively. In the present study thin layer chromatography and UV spectrophotometric method was established as a sensitive, economical and accurate method of detection and estimation of Camptothecin. The method was developed on TLC aluminum plates (60F from E-Merck Ltd.) precoated with silica gel using solvent system ethyl acetate: methanol (7.5:2.5, v/ v) which gives a dark blue spot of Camptothecin at 254 nm (R f value 0.46 cm). Spectrometric analysis of Camptothecin was carried out at the absorbance of 254 nm. Leaves and stems of Nothapodytes foetida and Ophiorrhiza mungos were extracted, subjected to TLC and UV spectrophotometric analysis and maximum absorption of CPT in ethyl acetate was observed at 254 nm. The quantification by this method was comparable to data obtained by HPLC. The optimized method was simple, repeatable, precise and cost effective, useful when there is large number of crude samples. In the present study this method was used to screen high yielding lines of CPT producing plants. [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: The folklore or traditional therapy in southern India widely utilizes a plethora of local herbs to treat the patients challenged with snake venom. Despite the widespread implementation of antisera therapy, the local population of the country still relies on this century's old medicinal formulas mainly due to the cost effectiveness, lesser side effects and also its cultural acceptability. The present study aims to validate the neutralizing ability of one such traditionally acclaimed antidote Ophiorrhiza mungos root extract against Russell's viper (Daboia russelii) venom in the early developing chick embryos. Materials and methods: The disc impregnated with venom, root extract or the combination of both was placed on the yolk sac membrane preferably over the anterior blood vessel of 6th day chick embryo. The neutralization/inhibition of venom-induced lethality or hemorrhage was achieved by incubating venom and extract before being applied to the embryo. The membrane stabilizing properties of root extract was estimated by HRBC lysis method. The preliminary phytochemical analysis was done to assess the phyto constituents in the root extract. Results: The LD50 of Russell's viper venom in 6th day chick embryo was found to be 3μg/μl. The neutralising effect of root extract was achieved by pre-incubating venom with various concentrations of extract and at the concentration of 10μg/μl, 100% recovery of embryos was observed after 6h of incubation. Higher concentration of root extract showed remarkable results by completely abolishing traces of hemorrhagic lesions induced by viper venom. Conclusions: The above observations confirmed that the root extract of Ophiorrhiza mungos possess potent anti snake venom neutralizing compounds, which inhibit the activity of viper venom. The chick embryo, a new insensate model used in the present study is significant in venom research as it reduces the ruthless suffering of higher mammalian experimental models. [ABSTRACT FROM AUTHOR]

Objective: The goal of this study was to evaluate the antioxidant and antiproliferative activities of 10 traditional medicinal plants, Asclepias curassavica, Ophiorrhiza mungos Linn., Cynodon dactylon (L.) Pers, Costus speciosus (J. Koenig.) Smith Costaceae, Achyranthes aspera L., Amaranthus tristis Roxb., Blepharis maderaspatensis L., Merremia emerginata Hall.f., Aegle marmelos Corr., and Tabernaemontana heyneana Wall., used in the traditional Indian system of medicine as a cure for cancer. The present study focuses on the anticancer potential of traditional medicinal plants to induce apoptosis in cancer cell lines. Methods: Plants were sequentially extracted with hexane, ethyl acetate, and methanol. The extract was concentrated to yield the crude extract, which was tested for antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl, nitric oxide and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays on four cancer cell lines and a normal cell line. The anticancer potential of cytotoxic extracts was determined by the Annexin-fluorescein isothiocyanate-conjugated assay in human colon adenocarcinoma cell lines (COLO 320 DM). Results: All the tested extracts showed significant antioxidant and antiproliferative activities in a concentration- and time-dependant manner in the following descending order: A. curassavica > C. dactylon > C. speciosus root > A. tristis > M. emarginata > O. mungos > T. Heyneana > B. maderaspatensis > A. marmelos > A. aspera. Conclusion: The results of the present study support the need of further studies to isolate potential anticancer drug with cancer cell-specific cytotoxicity. Additionally, the study supports the anticancer property of medicinal plants used in the traditional Indian medicine system and further evaluation of the selected medicinal plants for an effective anticancer drug with minimal side effects. [ABSTRACT FROM AUTHOR]

Camptothecin is an anticancer quinoline alkaloid effective against colon cancer. It acts by inhibition of the enzyme DNA topoisomerase I. A comparative study of camptothecin from the indigenous plants namely Nothapodytes foetida, Ophiorrhiza mungos and Ophiorrhiza rugosa indicated highest yields of camptothecin and 9-methoxy camptothecin in N. foetida. The other two plants O. mungos and O. rugosa contained low levels of alkaloids. [ABSTRACT FROM AUTHOR]

Camptothetin (CPT) is a quinoline alkaloid originally isolated from the Chinese tree, Camptotheca acuminata Decne. CPT was found to have anticancerous and antiviral properties. Derivatives of natural CPT, including topothecan and irinotecan are used clinically to treat a variety of cancers. Apart from Camptotheca acuminata Decne, CPT production was also found in the perennial plant Ophiorrhiza mungos. In this study we attempted the immobilization of the tissue culture grown callus of Ophiorrhiza mungos for the continuous production of a higher concentration of CPT. As evident from previous studies about the antiviral effects of CPT, we wanted to bioinformatically analyze the binding potential of CPT towards two important proteins of SARS-CoV-2, protease (Mpro) and RNA dependent RNA polymerase (RdRp). Further docking analysis of the CPT against the exterior spike glycoprotein of SARS-CoV-2 was also done to determine their potential interaction. The immobilized callus of Ophiorrhiza mungos produced CPT at a concentration of 420 µg/l by the end of 12 days of growth. The HPLC analysis was done to determine the purity of the CPT synthesized by the immobilization technique. The bioinformatic analysis revealed a higher binding efficiency of CPT and its derivatives, toptecan and irinotecan against Mpro and RdRp. The docking analysis of CPT against the spike glycoprotein of SARS-CoV-2 showed hydrogen bonding with the amino acids at K466 with a bond distance of 2.56A° and K355 with a bond distance of 2.40A°. This finding was of particular importance that other compounds including hydroxychloroquine sulphate, lopinavir and ivermectin could bind with the spike protein only by weak Vander wall bonds and no hydrogen bond formation was noticed. Our studies hence evaluate the efficiency of CPT against SARS-CoV-2, by potentially blocking the interaction of the spike glycoprotein with the angiotensin-converting enzyme 2 (ACE2) receptor found on host cells. [ABSTRACT FROM AUTHOR]

Vivipary, the precocious germination of seeds within the parent plant, is a specialised feature of evolutionary and biological importance that ensures survival of a plant. Reports on vivipary in angiosperms are rare, accounting for <0.1% of flowering plants. Here, we report a remarkable case of occurrence of vivipary in Ophiorrhiza mungos. A study was conducted to collect information on the morphology of the capsules that support vivipary, environmental factors that induce vivipary, survival mode and the survival of viviparous seedlings. The hydroscopic movement of the cup-shaped capsules of O. mungos was found to help in viviparous germination during the rainy season. Of the total seeds in a capsule, 70% showed viviparous germination. The seedlings remaining inside the capsule attain a height of 0.98 ± 0.4 cm and reach the ground when the capsule falls. On the ground, seedlings obtain easy anchorage to the substratum since they have already germinated. Vivipary appears to be an adaptation of O. mungos to the rainy season for ensuring viable offspring. This suggests that vivipary in this species might be artificially induced by continuous spraying with water to rescue seeds in all seasons for use in large-scale propagation to meet increasing market demand and conservation of this valuable anticancer medicinal herb. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To carry out the qualitative and quantitative evaluation of camptothecin, estimation of total phenolic compounds and evaluation of in vitro antioxidant activity and cytotoxic activity of Ophiorrhiza prostrata and Ophiorrhiza mungos. Methods: Direct Analysis in Real Time- Mass Spectrometry (DART-MS) was employed for the detection of camptothecin in the Ophiorrhiza species, while high performance thin layer chromatography (HPTLC) was used for the estimation of camptothecin. Total phenolic compounds were estimated by modified Folins-Ciocalteu''s reagent method. Antioxidant activity was evaluated through DPPH radical, hydroxyl radical, superoxide radical scavenging assays and reducing power assay. The cytotoxicity evaluation was performed using MTT assay on MCF-7 cell lines. Results: The presence of camptothecin was confirmed in both the species by the [M++H] peak at 349 by DART-MS analysis. Camptothecin content was estimated as 1.47 μg/gm (dry wt) in O. prostrata and 188.60 μg/gm (dry wt) in O. mungos using HPTLC method. The moderate in vitro antioxidant activities of the methanol extracts corroborates with the low content of phenolic compounds in O. prostrata (9.88 GAE mg/g) and O. mungos (12.73 GAE mg/g). The methanol extract of O. prostrata exhibited remarkable cytotoxicity on human breast cancer cell lines (MCF-7), with IC50 value 1.10μg/mL compared to O. mungos (3.48μg/mL) and standard camptothecin (3.51μg/mL). Conclusions: The application of DART-MS proved to be a simple and rapid technique for the detection of camptothecin in Ophiorrhiza species. The higher cytotoxicity for O. prostrata, despite the low content of camptothecin suggests the presence of other potential cytotoxic compounds in O. prostrata. [Copyright &y& Elsevier]