ArbuGuard Capsules (Supplement for Cancer)


  • ArbuGuard is an herbal dietary supplement act as an immunomodulator, anti-oxidant, anti-bacterial, anti-cancerous, digestive, nutritive and anti-aging in property.
  • Vegetarian Product
  • 100% herbal, no side effects reported
  • Clinically proven, safe in pregnancy and lactation
  • Doage – 1-2 capsules twice a day before or after food.


 “ArbuGuard” is a poly-herbal formulation made-up of aqueous extracts of 8 wonder herbs. These herbs are well-known and widely used in India and other territories in food or traditional medicine. “Arbud” is a Sanskrit word which means a tumor, swelling, cyst or cancer.

Ingredients of “ArbuGuard”: Each HPMC (Veg) capsule contains aq. Extract of –

  1. Trichosanthes dioica (Patola) – 95 mg
  2. Terminalia chebula (Haritaki) – 95 mg
  3. Terminalia bellirica (Vibhitaki)  – 95 mg
  4. Emblica officinale/ Phyllanthus emblica (Amalaki) – 95 mg
  5. Cyprus rotundus (Mustaka)  – 95 mg
  6. Azadirecta indica (Nimba) – 95 mg
  7. Holarrhena antidysenterica (Kutaj) – 95 mg
  8. Haridra (Curcuma longa) – 35 mg

Dosage – 1-2 capsules twice a day before/ after food.

Availability – 60 veg capsules in a HDPE container

Various research studies on these ingredients proved that each ingredient has their specific properties and effects on human body. A number of research papers were published in journal which showed positive health benefits of these ingredients. They are immunomodulator, anti-oxidant, anti-bacterial, anti-cancerous, digestive, nutritive and anti-aging in property. After studying Ayurvedic classical texts and reviewing various drug research publications, we formulated “ArbuGuard” and promoting it as an herbal dietary supplement for Cancer patient. We also found that it helps in proper digestion of food and excretion of waste. It is also very useful in reduction of pile mass (anti-haemorrhoidal effect), lipoma, cyst and deep scars of acne vulgaris. Though, there are a lot of research work done on ingredients of “ArbuGuard”, we are highlighting only anti-cancer activities below.

Anti-cancer activity of Trichosanthes dioica:

Trichosanthes dioica at the doses of 5 and 10 mg/kg demonstrated marked antitumor effect as evidenced by significant reduction in tumor volume, tumor weight, packed cell volume, viable tumor cell count and increase in normal peritoneal cell count, non-viable tumor cell count, MST and life span of tumor bearing hosts. Trichosanthes dioica significantly restored the altered hematological parameters toward normal values. The Trichosanthes dioica also significantly recuperated the hepatic antioxidant parameters, viz., lipid peroxidation, reduced GSH level, activities of GST, SOD and CAT in tumor bearing mice.

(Ref. Antitumor efficacy and amelioration of oxidative stress by Trichosanthes dioica root against Ehrlich ascites carcinoma in mice – Sanjib Bhattacharya,Angelene Prasanna,Piyali Majumdar,RB Suresh Kumar & Pallab K. Haldar; link to article –

Anti-cancer activity of Terminalia chebula:

  1. A 70% methanol extract of Terminalia chebula fruit, was studied for its effects on growth in several malignant cell lines including a human (MCF-7) and mouse (S115) breast cancer cell line, a human osteosarcoma cell line (HOS-1), a human prostate cancer cell line (PC-3) and a non-tumorigenic, immortalized human prostate cell line (PNT1A) using assays for proliferation ([3H]-thymidine incorporation and coulter counting), cell viability (ATP determination) and cell death (flow cytometry and Hoechst DNA staining). In all cell lines studied, the extract decreased cell viability, inhibited cell proliferation, and induced cell death in a dose dependent manner. Flow cytometry and other analyses showed that some apoptosis was induced by the extract at lower concentrations, but at higher concentrations, necrosis was the major mechanism of cell death. ATP assay guided chromatographic fractionation of the extract yielded ellagic acid, 2,4-chebulyl-β-d-glucopyranose (a new natural product), and chebulinic acid which were tested by ATP assay on HOS-1 cell line in comparison to three known antigrowth phenolics of Terminalia, gallic acid, ethyl gallate, luteolin, and tannic acid. Chebulinic acid (IC50=53.2 μM±0.16)>tannic acid (IC50=59.0 μg/ml±0.19)> and ellagic acid (IC50=78.5 μM±0.24), were the most growth inhibitory phenolics of T. chebula fruit in our study. (Ref.
  2. Evaluation of anticancer potential of Terminalia chebula Fruits against Ehrlich Ascites Carcinoma induced cancer in mice – Rohini Ahuja, Neeraj Agrawal, Alok Mukerjee – This study was designed to determine the in vivo and in vitro anticancer potential of the ethanolic extract of Terminalia chebula (ETC) fruits against Ehrlich Ascites Carcinoma (EAC) induced cancer in swiss albino mice. The anticancer activity was assessed using in vitro cytotoxicity, mean survival time, tumor volume and hematological studies. The reliable criteria for evaluating the potential of any anticancer agent is the prolongation of lifespan of the animal and decrease in WBC count of blood. The high dose of ETC (200 mg/kg, orally) significantly reduced the tumor growth which was demonstrated by increased lifespan of the mice and restoration of hematological parameters. ETC was also found to be cytotoxic in the in vitro parameter which shows that ETC possesses significant anticancer potential. (ref.
  3. Aqueous Extract of Terminalia chebula Induces Apoptosis in Lung Cancer Cells Via a Mechanism Involving Mitochondria-mediated Pathways – The current study was designed to evaluate the aqueous extract of Terminalia chebula activity, and the main pathway was detected on lung cancer by extracts of T. chebula. Aqueous extract of T. chebula was separated using a zeolite, and five fractions of T. chebula extract were obtained and analyzed by ultraviolet (UV) and infrared (IR) spectroscopy. Antiproliferative activity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) methods against human lung cancer (A549) and mouse lung cancer cell line LLC. T. chebula acts by regulating the Bcl-2 family protein-mediated mitochondrial pathway detected by western blot. Fraction 4 of the chebula extract showed much function and was thus studied further. Fraction 4 increased the activation of caspase-3, induced PARP cleavage, and promoted cytochrome c release into the cytoplasm. These data suggest that T. chebula acts by regulating the Bcl-2 family protein-mediated mitochondrial pathway and provide evidence that T. chebula deserves further investigation as a natural agent for treating and preventing cancer. (ref.
  4. Anti-colon cancer activity of endophytic fungal strains from Terminalia chebula – Endophytic microorganisms are fungi or bacteria that live inside the healthy tissues of the host plants causing no apparent symptoms of diseases. Five endophytic fungal strains labeled as IR-1, IR-2, IR-4, IR-6 and IR-7 (identified as Penicillium thiomii) were isolated from the medicinal plant of Terminalia chebula Retz by culture and sub-culture. The ethyl acetate extract of fungal strains, IR-4, IR-6 and IR-7 inhibited the growth of CaCo-2 colon cancer cell lines in MTT assay with IC50 values of 55, 44 and 67 µg/mL respectively. (ref.

Anti-cancer activity of Terminalia bellirica –

  1. Terminalia bellirica extract induces anticancer activity through modulation of apoptosis and autophagy in oral squamous cell carcinoma – Terminalia bellirica (TB) has been used in traditional Indian medical system, Ayurveda. However, the mechanism underlying the efficacy of the TB extract against oral squamous cell carcinoma (OSCC) is yet to be explored. The present study established a connecting link between the TB extract induced apoptosis and autophagy in relation to reactive oxygen species (ROS). Our study revealed, that gallic acid in the TB extract possess a strong free radical scavenging capacity contributing towards the selective anti-proliferative activity. Furthermore, TB extract markedly enhanced the accumulation of ROS that facilitated mitochondrial apoptosis through DNA damage, indicating ROS as the vital component in regulation of apoptosis. This effect was effectively reversed by the use of a ROS scavenger, N-acetyl cysteine (NAC). Moreover, it was observed to induce autophagy; however, it attenuated the autophagosome-lysosome fusion in Cal33 cells without altering the lysosomal activity. Pharmacological inhibitors of autophagy, namely, 3-methyladenine and chloroquine, were demonstarated to regulate the stage-specific progression of autophagy post treatment with the TB extract, favouring subsequent activation of apoptosis. These findings revealed, presence of gallic acid in TB extract below NOAEL value causes oxidative upset in oral cancer cells and promote programmed cell death which has a potential therapeutic value against oral squamous cell carcinoma. ( ref.
  2. Octyl gallate and gallic acid isolated from Terminalia bellarica regulates normal cell cycle in human breast cancer cell lines – Herbal medicines stand unique and effective in treating human diseases. Terminalia bellarica (T. bellarica) is a potent medicinal herb, with a wide range of pharmacological activities. The present study was aimed to evaluate the effect of octyl gallate (OG) and gallic acid (GA) isolated from methanolic fruit extract of T. bellirica to inhibit the survival of breast cancer cells (MCF-7 & MDA-MB-231). Both OG & GA exhibited decreased MCF-7 & MDA-MB-231 survival and induced apoptosis, with IC50 value of OG and GA as 40 μM and 80 μM respectively. No toxic effect was observed on normal breast cells (MCF-10A). The compounds inhibited cell cycle progression by altering the expression of the cell cycle regulators (Cyclin D1, D3, CDK-4, CDK-6, p18 INK4, p21Waf-1 and p27 KIP). Octyl gallate was more effective at low concentrations than GA. In-silico results provided stable interactions between the compounds and target proteins. The present investigation proved the downregulation of positive cell cycle regulators and upregulation of negative cell cycle regulators inducing apoptosis in compound-treated breast cancer cells. Hence, both the compounds may serve as potential anticancer agents and could be developed as breast cancer drugs, with further explorations. (ref.

Anti-cancer activity of Emblica officinale/ Phyllanthus emblica

  1. Antitumour effects of phyllanthus emblica L.: Induction of cancer cell apoptosis and Inhibition of in vivo tumour promotion and in vitro invasion of human cancer cells – Phyllanthus emblica Linn. (PE) is a medicinal fruit used in many Asian traditional medicine systems for the treatment of various diseases including cancer. The present study tested the potential anticancer effects of aqueous extract of PE in four ways: (1) against cancer cell lines, (2) in vitro apoptosis, (3) mouse skin tumourigenesis and (4) in vitro invasiveness. The PE extract at 50–100 µg/mL significantly inhibited cell growth of six human cancer cell lines, A549 (lung), HepG2 (liver), HeLa (cervical), MDA‐MB‐231 (breast), SK‐OV3 (ovarian) and SW620 (colorectal). However, the extract was not toxic against MRC5 (normal lung fibroblast). Apoptosis in HeLa cells was also observed as PE extract caused DNA fragmentation and increased activity of caspase‐3/7 and caspase‐8, but not caspase‐9, and up‐regulation of the Fas protein indicating a death receptor‐mediated mechanism of apoptosis. Treatment of PE extract on mouse skin resulted in over 50% reduction of tumour numbers and volumes in animals treated with DMBA/TPA. Lastly, 25 and 50 µg/mL of PE extract inhibited invasiveness of MDA‐MB‐231 cells in the in vitro Matrigel invasion assay. These results suggest P. emblica exhibits anticancer activity against selected cancer cells, and warrants further study as a possible chemopreventive and antiinvasive agent. (
  2. Emblica officinalis Extract Induces Autophagy and Inhibits Human Ovarian Cancer Cell Proliferation, Angiogenesis, Growth of Mouse Xenograft Tumors – Patients with ovarian cancer (OC) may be treated with surgery, chemotherapy and/or radiation therapy, although none of these strategies are very effective. Several plant-based natural products/dietary supplements, including extracts from Emblica officinalis (Amla), have demonstrated potent anti-neoplastic properties. In this study we determined that Amla extract (AE) has anti-proliferative effects on OC cells under both in vitro and in vivo conditions. We also determined the anti-proliferative effects one of the components of AE, quercetin, on OC cells under in vitro conditions. AE did not induce apoptotic cell death, but did significantly increase the expression of the autophagic proteins beclin1 and LC3B-II under in vitro conditions. Quercetin also increased the expression of the autophagic proteins beclin1 and LC3B-II under in vitro conditions. AE also significantly reduced the expression of several angiogenic genes, including hypoxia-inducible factor 1α (HIF-1α) in OVCAR3 cells. AE acted synergistically with cisplatin to reduce cell proliferation and increase expression of the autophagic proteins beclin1 and LC3B-II under in vitro conditions. AE also had anti-proliferative effects and induced the expression of the autophagic proteins beclin1 and LC3B-II in mouse xenograft tumors. Additionally, AE reduced endothelial cell antigen – CD31 positive blood vessels and HIF-1α expression in mouse xenograft tumors. Together, these studies indicate that AE inhibits OC cell growth both in vitro and in vivo possibly via inhibition of angiogenesis and activation of autophagy in OC. Thus AE may prove useful as an alternative or adjunct therapeutic approach in helping to fight OC. (Ref.
  3. Emblica officinalis extract downregulates pro-angiogenic molecules via upregulation of cellular and exosomal miR-375 in human ovarian cancer cells – Ovarian cancer (OC) is highly resistant to current treatment strategies based on a combination of surgery, chemotherapy and radiation therapy. We have recently demonstrated the anti-neoplastic effect of Amla extract (Emblica officinalis, AE) on OC cells in vitro and in vivo. We hypothesized that AE attenuates growth of OC through microRNA (miR)-regulated mechanism(s). The inhibitory effect of AE on proliferation, migration and invasiveness (P≤0.001) of SKOV3 cells and >90% attenuation of tumor growth in a xenograft mouse model suggested multiple targets. RT-qPCR analysis of microRNAs associated with OC showed a >2,000-fold increase in the expression of miR-375 in AE-treated SKOV3 cells that was blocked by an exogenous miR-375 inhibitor (P≤0.001). AE also decreased the gene and protein expression of IGF1R, a target of miR-375 (P≤0.001), and SNAIL1 (P≤0.002), an EMT-associated transcription factor that represses E-cadherin expression (P≤0.003). AE increased E-cadherin expression (P≤0.001). Treatment of SKOV3 cells with AE resulted in increased miR-375 in exosomes in the medium (P≤0.01). Finally, AE significantly decreased the expression of IGF1R and SNAIL1 proteins during attenuation of SKOV3-derived xenograft tumor. Together, these results show that AE modulates cancer cells and the tumor microenvironment via activation of miR-375 and by targeting IGF1R and SNAIL1 in OC cells. (Ref.
  4. Anticancer Activity of Phyllanthus emblica Linn. (Indian Gooseberry): Inhibition of Transcription Factor AP-1 and HPV Gene Expression in Cervical Cancer Cells – Plant products of Phyllanthus emblica Linn. are traditionally consumed for its immense nutritive and medicinal values. However, the molecular mechanism(s) by which it exerts it effects is less understood. In this study, we investigated mechanism of action of P. emblica fruit extract (PE) by studying its effect on activator protein-1 (AP-1) activity and human papillomavirus (HPV) transcription that are essential for tumorigenicity of cervical cancer cells. PE resulted in a dose-and time-dependent inhibition of DNA binding activity of constitutively active AP-1 in both HPV16-positive (SiHa) and HPV18-positive (HeLa) cervical cancer cells. PE-induced AP-1 inhibition was found mediated through downregulation of constituent AP-1 proteins, c-Jun, JunB, JunD, and c-Fos; however, the kinetics of their inhibition varied in both the cell types. Inhibition of AP-1 by PE was accompanied by suppression of viral transcription that resulted in growth inhibition of cervical cancer cells. Growth inhibitory activity of PE was primarily manifested through induction of apoptotic cell death. These results suggest that P. emblica exhibits its anticancer activities through inhibition of AP-1 and targets transcription of viral oncogenes responsible for development and progression of cervical cancer thus indicating its possible utility for treatment of HPV-induced cervical cancers. (ref.
  5. Indian gooseberry (Emblica officinalis Gaertn.) suppresses cell proliferation and induces apoptosis in human colon cancer stem cells independent of p53 status via suppression of c-Myc and cyclin D1 – Indian gooseberry, also known as amla, a widely consumed fruit in South Asia, was evaluated for its anti-proliferative and pro-apoptotic mechanisms on human colon cancer stem cells (HCCSC). Amla extracts suppressed proliferation and induced apoptosis independent of p53, a tumour suppressor gene, in HCCSCs. Further, amla extracts suppressed cell proliferation by targeting the Wnt/β-catenin signalling pathway as seen by decreased nuclear translocation of β-catenin. Additionally, this led to suppressed expression of c-Myc and cyclin D1, key proteins involved in cell proliferation. Inhibition of stem-ness of HCCSCs by amla may be due to its effect on the Wnt/β-catenin signalling. These results indicate that amla suppresses HCCSC proliferation and induces apoptosis independent of p53 status via potentially targeting Wnt/β-catenin signalling pathway. Amla is therefore a promising functional food for preventing colon cancer and might be a novel resource for the food industry. (ref.

Anti-cancer activity of Cyprus rotundus –

  1. Cytotoxic effect of Cyperus rotundus rhizome extract on human cancer cell lines – The wild weed Cyperus rotundus is commonly used as traditional medicine in different parts of the world. Sequential extraction of C. rotundus rhizome with solvents of different polarity namely hexane, chloroform, ethyl acetate, methanol and water were prepared and the free radical scavenging activity was determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Based on high antioxidant activity of methanolic extract of C. rotundus rhizome (MRCr) was further investigated for its cytotoxic effect on different human cancer cell lines-breast (MCF-7), cervical (HeLa), liver (Hep G2), prostate (PC-3), colorectal (HT-29) and normal cell line (MCF-12A) by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay evaluated as 50% inhibition of growth (IC50). Apoptosis cells were analysed by flow cytometry stained with annexin V-Fluorescein isothiocyanate conjugate (AF) and propidium iodide (PI). The cellular and nuclear changes were examined under light and fluorescent microscope using 4′, 6′ diamino-2-phenylindole (DAPI) stain, dual stains of AF/PI and acridine orange/ethidium bromide (AO/EB). The cytotoxic effects on the tested cancer cell lines ranged from 4.52 ± 0.57 to 9.85 ± 0.68 μg ml−1. The migration assay was showed the inhibitory effect with MRCr. The MRCr showed significant anticancer activity against all the tested cancer cell lines and also protected the non-cancer cells. The anticancer activity suggests further elucidation for the formulation of natural pharmaceutical products in the treatment of cancer. (ref.
  2. 6‐Acetoxy Cyperene, a Patchoulane‐type Sesquiterpene Isolated from Cyperus rotundus Rhizomes Induces Caspase‐dependent Apoptosis in Human Ovarian Cancer Cells – Cyperus rotundus (Cyperaceae) has been widely used in traditional medicine for the treatment of various diseases, including cancer. Although an anti‐tumour effect has been suggested for C. rotundus, the anti‐tumour effects and underlying molecular mechanisms of its bioactive compounds are poorly understood. The n‐hexane fraction of an ethanol extract of C. rotundus rhizomes was found to inhibit cell growth in ovarian cancer (A2780, SKOV3 and OVCAR3) and endometrial cancer (Hec1A and Ishikawa) cells. Among the thirteen sesquiterpenes isolated from the n‐hexane fraction, some patchoulane‐type compounds, but not eudesmane‐type compounds, showed moderate cytotoxic activity in human ovarian cancer cells. In particular, the patchoulane sesquiterpene 6‐acetoxy cyperene had the most potent cytotoxicity. In this regard, propidium iodide/Annexin V staining and terminal deoxynucleotidyl transferase dUTP (deoxynucleotide triphosphate) nick end labeling assay were performed to study cell cycle progression and apoptosis. 6‐acetoxy cyperene induced apoptosis, as shown by the accumulation of sub‐G1 and apoptotic cells. Furthermore, treatment with 6‐acetoxy cyperene stimulated the activation of caspase‐3, caspase‐8 and caspase‐9 and poly(ADP‐ribose)polymerase in a dose‐dependent manner. Pretreatment with caspase inhibitors neutralized the pro‐apoptotic activity of 6‐acetoxy cyperene. Taken together, these data suggest that 6‐acetoxy cyperene, a patchoulane‐type sesquiterpene isolated from C. rotundus rhizomes, is an anti‐tumour compound that causes caspase‐dependent apoptosis in ovarian cancer cells. (ref.
  3. The treatment role of Cyperus rotundus L. to triple-negative breast cancer cells – Cyperus rotundus L. is widely used in Traditional Chinese Medicine and studies have reported its anticancer effect, but its chemical composition and therapy mechanism remains unknown. This research aims to analyze the chemical components of the ethanol extract of Cyperus rotundus L. (EECR), detect its treatment effects on human Triple-negative breast cancer (TNBC) cells, and elucidate possible therapy mechanisms. The chemical components of EECR were detected by the Waters UPLC combined with Bruker Q-TOF mass spectrometer (UPLC-Q-TOF-MS). The phytochemical compounds were identified by comparing the mass fragmentations of each metabolite with databases such as METLIN, HMDB, and NCBI. A total of 21 compounds were identified in EECR. MDA-MB-231 and MDA-MB-468 cells were treated with various concentrations of EECR. Cell proliferation was examined using Cell Counting Kit-8 (CCK-8) and colony formation assays. Cell apoptosis and cell cycle were detected by flow cytometry. Apoptosis- and autophagy-related protein expression was detected by Western blot. EECR inhibits the proliferation of TNBC cells (MDA-MB-231 and MDA-MB-468) in a dose-dependent manner, which may be related to the arrest of cell cycle in G0/G1 phase. It induces apoptosis by promoting the expression of BAX and inhibiting the expression of BCL-2. In addition, autophagy inhibitor 3-Methyladenine (3-MA) inhibited TNBC cells pro-survival autophagy and increased the sensitivity of EECR. The present results demonstrated that EECR has potential effects on inhibits the proliferation and induction apoptosis in TNBC. (ref.
  4. Activity Anticancer n-hexane Fraction of Cyperus Rotundus l. Rhizome to Breast Cancer MCF-7 Cell Line – The fraction n-hexane Cyperus rutundus L. rhizome has anticancer activity against breast cancer MCF-7 cells with accumulation cell cycle in the G0-G1 phase and through induction of apoptosis. (ref.
  5. The Ethanol Crude Extraction of Cyperus Rotundus Regulates Apoptosis-associated Gene Expression in HeLa Human Cervical Carcinoma Cells In Vitro – The treatment of CRE on HeLa cells caused morphological changes and induced chromatin condensation. DNA microarray analysis showed that CRE led to up-regulation of 449 genes and down-regulation of 484 genes, which were classified in several interaction pathways. Conclusion: CRE changed HeLa cell morphology and induced gene expression which associated with apoptosis and cell-cycle arrest. These results provide important information at the transcription level for targeting treatments of human cervical cancer. (ref.
  6. In vitro evaluation of antibacterial, antioxidant, cytotoxic and apoptotic activities of the tubers infusion and extracts of Cyperus rotundus – The in vitro antibacterial, antioxidant, cytotoxic and apoptotic activities from tubers extracts of Cyperus rotundus (Cyperaceae) were investigated. Antibacterial activity of different extracts was evaluated against five bacterial reference strains. A marked inhibitory effect was observed against Salmonella enteritidis, Staphylococcus aureus and Enterococcus faecalis with total oligomers flavonoids (TOFs) and ethyl acetate extracts. In addition to their antibacterial activity, the same extracts showed a significant ability to inhibit nitroblue tetrazolium reduction by the superoxide radical in a non-enzymatic superoxide generating system. Apoptosis, a highly organized physiological mechanism to eliminate injured or abnormal cells, is also implicated in multistage carcinogenesis. It was observed that TOF and ethyl acetate extracts suppressed growth and proliferation of L1210 cells derived from murine lymphoblastic leukaemia. Morphological features of treated cells and characteristic DNA fragmentation revealed that the cytotoxicity was due to induction of apoptosis. This study confirms that TOF and ethyl acetate extracts of C. rotundus possess antibacterial and antioxidant properties and provoke DNA fragmentation, a sign of induction of apoptosis. These results were correlated with chemical composition of the tested extracts. (ref.

Anti-cancer activity of Azadirecta indica –

  1. Antiproliferative effect on human cancer cell lines after treatment with nimbolide extracted from an edible part of the neem tree (Azadirachta indica) – Nimbolide, a triterpenoid extracted from the flowers of the neem tree (Azadirachta indica), was found to have antiproliferative activity against some cancer cell lines. Treatment of cells with 0.5–5.0 μm concentrations of nimbolide resulted in moderate to very strong growth inhibition in U937, HL‐60, THP1 and B16 cell lines. Flow cytometric analysis of U937 cells showed that nimbolide treatment (1–2.5 μm) resulted in cell cycle disruption by decreasing the number of cells in G0/G1 phase, with initial increases in S and G2/M phases. Cells exposed to a higher dose of nimbolide for a longer period displayed a severely damaged DNA profile, resulting in a remarkable increase in the number of cells in the sub‐G1 fraction, with a reciprocal decrease of cells in all phases. Quantification of the expression of phosphatidylserine in the outer cell membrane showed that doses of nimbolide higher than 0.4 μm exerted remarkable lethality, with over 60% of cells exhibiting apoptotic features after exposure to 1.2 μm nimbolide. The antiproliferative effect of nimbolide and its apoptosis‐inducing property raise hope for its use in anticancer therapy by enhancing the effectiveness of cell cycle disruption.(ref.
  2. Novel Molecular Targets of Azadirachta indica Associated with Inhibition of Tumor Growth in Prostate Cancer – Advanced prostate cancer has significant long-term morbidity, and there is a growing interest in alternative and complimentary forms of therapy that will improve the outcomes of patients. Azadirachta indica (common name: neem) contains multiple active compounds that have potent anti-inflammatory and anticancer properties. The present study investigates the novel targets of the anticancer activity of ethanol extract of neem leaves (EENL) in vitro and evaluates the in vivo efficacy in the prostate cancer models. Analysis of the components in the EENL by mass spectrometry suggests the presence of 2′,3′-dehydrosalannol, 6-desacetyl nimbinene, and nimolinone. Treatment of C4-2B and PC-3M-luc2 prostate cancer cells with EENL inhibited the cell proliferation. Genome-wide expression profiling, using oligonucleotide microarrays, revealed genes differentially expressed with EENL treatment in prostate cancer cells. Functional analysis unveiled that most of the up-regulated genes were associated with cell death, and drug metabolism, and the down-regulated genes were associated with cell cycle, DNA replication, recombination, and repair functions. Quantitative PCR confirmed significant up-regulation of 40 genes and immunoblotting revealed increase in the protein expression levels of HMOX1, AKR1C2, AKR1C3, and AKR1B10. EENL treatment inhibited the growth of C4-2B and PC-3M-luc2 prostate cancer xenografts in nude mice. The suppression of tumor growth is associated with the formation of hyalinized fibrous tumor tissue and the induction of cell death by apoptosis. These results suggest that EENL-containing natural bioactive compounds could have potent anticancer property and the regulation of multiple cellular pathways could exert pleiotrophic effects in prevention and treatment of prostate cancer. (ref.
  3. Preclinical Evaluation of the Supercritical Extract of Azadirachta Indica (Neem) Leaves In Vitro and In Vivo on Inhibition of Prostate Cancer Tumor Growth – Azadirachta indica, commonly known as neem, has gained worldwide prominence because of its medical properties, namely antitumor, antiviral, anti-inflammatory, antihyperglycemic, antifungal, and antibacterial activities. Despite these promising results, gaps remain in our understanding of the molecular mechanism of action of neem compounds and their potential for use in clinical trials. We investigated supercritical extract of neem leaves (SENL) for the following: molecular targets in vitro, in vivo efficacy to inhibit tumor growth, and bioactive compounds that exert antitumor activity. Treatment of LNCaP-luc2 prostate cancer cells with SENL suppressed dihydrotestosterone-induced androgen receptor and prostate-specific antigen levels. SENL inhibited integrin β1, calreticulin, and focal adhesion kinase activation in LNCaP-luc2 and PC3 prostate cancer cells. Oral administration of SENL significantly reduced LNCaP-luc2 xenograft tumor growth in mice with the formation of hyalinized fibrous tumor tissue, reduction in the prostate-specific antigen, and increase in AKR1C2 levels. To identify the active anticancer compounds, we fractionated SENL by high-pressure liquid chromatography and evaluated 16 peaks for cytotoxic activity. Four of the 16 peaks exhibited significant cytotoxic activity against prostate cancer cells. Mass spectrometry of the isolated peaks suggested the compounds with cytotoxic activity were nimbandiol, nimbolide, 2′,3′-dihydronimbolide, and 28-deoxonimbolide. Analysis of tumor tissue and plasma samples from mice treated with SENL indicated 28-deoxonimbolide and nimbolide as the bioactive compounds. Overall, our data revealed the bioactive compounds in SENL and suggested that the anticancer activity could be mediated through alteration in androgen receptor and calreticulin levels in prostate cancer. Mol Cancer Ther; 13(5); 1067–77. ©2014 AACR. (ref.
  4. Azadirachta indica exhibits chemopreventive action against hepatic cancer: Studies on associated histopathological and ultrastructural changes – The present study was designed to evaluate the anticarcinogenic potential of Azadirachta indica against N‐nitrosodiethylamine (NDEA)‐induced hepatocarcinogenesis. Further, the associated histopathological and ultrastructural changes were also analyzed. Hepatic cancer model was developed by the intraperitoneal administration of NDEA to mice at weekly intervals, in successive increasing doses, for a period of 8 weeks. Aqueous A. indica leaf extract (AAILE) was administered orally at a dosage of 100 μg/g body weight thrice a week till termination of the study. A relationship between histopathological grading and chemopreventive effect of A. indica had been established at various stages of carcinogenesis. Anticancer activity of A. indica was evaluated in terms of tumor incidence, tumor multiplicity, and survival rate. A significant reduction in tumor incidence (33%), tumor multiplicity (42%), and increase in survival (34%) was observed upon administration of AAILE to NDEA‐abused mice. Transmission and scanning electron microscopic investigations showed severe alterations in organelle organization, cellular arrangement, degree of differentiation, cellular metabolism, and morphology of the hepatocytes. These changes appeared to be distinctly delayed upon AAILE supplementation. The results suggest A. indica may have anticancer potential against NDEA‐induced hepatic cancer. Microsc. Res. Tech., 2012. © 2011 Wiley Periodicals, Inc. (ref/
  5. Effect of alkaline pH on cytotoxicity profile of neem (Azadirachta indica) ethanolic extract against human breast cancer cell line MDA-MB-231 – Results revealed that an increase in alkaline pH reduced the growth of breast cancer cells. Moreover, combined treatment of neem extract and alkaline pH decreased the growth and survival of cancer cells in a dose and pH-dependent manner. Neem extract at a concentration of 1600 μg/mL and pH 8.6 caused significant mortality in breast cancer cells. A significant increase in antioxidant activity of neem extract was also observed as a function of both pH and dose of neem extract. In addition, the neem extract showed significant antibacterial activity against S. aureus at 200 mg/mL while no appreciable activity was detected against E. coli. (ref.

Anti-cancer property of Holarrhena antidysenterica –

  1. In vitro cytotoxic activity of leaves extracts of Holarrhena antidysenterica against some human cancer cell lines – In vitro cytotoxic potential of extracts (95% and 50% ethanolic extract and hot water extract at concentration of 100 µg/ml) from leaves of Holarrhena antidysenterica was evaluated against fourteen human cancer cell lines — A-549, COLO-205, DU-145, HeLa, HEP-2, IMR-32, KB, MCF-7, NCI-H23, OVCAR-5, SiHa, SK-N-MC, SW-620 and ZR-75-1 from nine different tissues (breast, colon, cervix, CNS, lung, liver, oral, ovary and prostate) using SRB assay. The 95% ethanolic extract displayed maximum anti-proliferative effect in the range of 73-92% against eight human cancer cell lines, while 50% ethanolic extract showed cytotoxic activity in the range of 70-94% against seven human cancer cell lines. However, the hot water extract did not show any activity. Among the fractions of 95% and 50% ethanolic extract, significant cytotoxic activity was found in the chloroform soluble fraction of 95% ethanolic extract at 100 µg/ml; it inhibited the growth in the range of 71-99% of seven human cancer cell lines from five different tissues viz., OVCAR-5 (ovary), HT-29 (colon), SK-N-MC (neuroblastoma), HEP-2 (liver), COLO-205 (colon), NIH-OVCAR-3 (ovary) and A-549 (lung). The cytotoxic activity of chloroform soluble fraction was found to be higher than 5-flurouracil, adriamycin, mitomycin-c and paclitaxel (anticancer drugs used as positive controls). Further in vivo studies and identification of active components from the chloroform fraction and their exact mechanism of action could be useful in designing new anticancer therapeutic agents. (ref.
  2. Induction of apoptosis in human bladder cancer cells by triterpenoids isolated from Holarrhena antidysenterica through differential reactive oxygen species generation – A novel triterpenoid, holarol(1),3β-lup-20(31)-en-3,29,30-triol along with one seco-triterpenoid, dihydrocanaric acid(2) and one known pentacyclic triterpenoid, betulin(3) have been isolated from Holarrhena antidysenterica (L.)Wall. (Family: Apocynaceae). The structures of the compounds were elucidated by extensive IR, 1D, 2D NMR and mass spectrometric analysis. The optimised geometry of (1) was calculated by density-functional theory (DFT) using M06-2X hybrid functional and 6–31 G(D) basis set. The compounds showed differential cytotoxic activities in the cell lines-HeLa, EAC, Raji and T24. Seco-triterpenoid (2) showed highest sensitivity (IC50: 1.710 μg/mL) against the bladder cancer cell line T24 followed by (1) (IC50 9.698 µg/mL) and (3) (IC50 11.769 µg/mL). Compound (1) showed highest reactive oxygen species (ROS) generation in T24 cell line followed by (3) and (2) resulting in induction of apoptosis through activation of caspase, cleavage of PARP and reduction of Bcl-2/Bax ratio. Thus compounds (1), (2) along with (3) could be potent anticancer agents. (ref.
  3. Methanol Extract of Holarrhena antidysenterica Inhibits the Growth of Human Oral Squamous Cell Carcinoma Cells and Osteoclastogenesis of Bone Marrow Macrophages – Oral squamous cell carcinoma (OSCC) frequently invades mandibular bone, and outcomes for treatment with surgical resection are typically poor, ultimately resulting in death. Holarrhena antidysenterica L. (Apocynaceae), distributed throughout Sri Lanka and India, has been used as a folk remedy to treat various diseases. Treatment with methanol extract of H. antidysenterica bark (HABE) inhibited cell viability and BrdU incorporation and induced apoptotic cell death in Ca9-22 gingival and HSC-3 tongue SCC cells. Flow cytometric analysis indicated that HABE treatment preferentially induces apoptotic cell death via increasing the sub-G1 peak in Ca9-22 cells and cell cycle arrest at the G1 phase in HSC-3 cells. HABE treatment in the presence of zVAD-fmk, a pan-caspase inhibitor, rescued cell viabilities in both OSCC cell lines. The ratio of Bax to Bcl-2 increased with reductions in the Bcl-2 protein expression, and the activation of caspase 3 and subsequent cleavage of PARP was detected in HABE-treated Ca9-22 and HSC-3 cells. Furthermore, HABE treatment at noncytotoxic concentrations inhibited osteoclast formation in RANKL-stimulated bone marrow macrophages. Taken together, HABE possesses the inhibitory activity on the growth of OSCC cells and antiosteoclastogenic activity. Therefore, HABE may be a promising alternative and complementary agent for preventing and treating OSCC. (ref.

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