Therapeutic use of plant extracts as nano-particles


Introduction with Problem Identification

Medicinal plants are considered as a warehouse of numerous types of bioactive compounds possessing wide-ranging therapeutic properties. The therapeutic potential of plants has been well explored over a very long time period. The enormous array of therapeutic effects associated with medicinal plants includes anti-cancer, anti-inflammatory, anti-viral, anti-tumor, anti-malarial, and analgesic. Cancer is one of the major problems to human health around the world. Cancer has been a ranked first due to mortality rates. The changing lifestyle of the population has been the lead cause. 23% of total cancer cases are breast cancer among females and 17% accounts for lung cancer among males. Lack of early diagnosis and treatment has resulted in decreased survival rate. (Raina H et al 2014, Jemal A et al 2011)

Current cancer therapy strategies are based on surgery, radiotherapy and chemotherapy. Chemotherapy shows greater efficiency in advanced stages of cancer treatment. Irrespective of the advancements in chemotherapy, there are many biological barriers that affect drug delivery to target cells. Some of the cancer stem cells (CSC) can self-renew, give rise to heterogeneous populations of daughter cells, and proliferate extensively. Multidrug resistance mechanisms by cancer cells lead to decrease in efficacy of the drug due to reduced drug concentration in target cells. (Sanna A et al 2014)

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So far many herbal medicines is been used as a potential therapeutics against cancer. Many studies indicate the different mechanisms of action of herbal medicines which mainly includes anti-oxidant properties, anti-tumorigenic properties and also suppressive action. (Alam S et al 2013) Many herbal extracts inhibits cell division process by break down of microtubules. Some herbal extracts provides the defense mechanism against cancer by enhancing cytotoxic activity of the monocytes and the macrophages. Some extracts such as Curcumin can suppress mutagenic effect of various mutagens.

Nano therapy aims in the targeted drug delivery to the cells. Nanoparticles are synthesized by biodegradable materials. It helps in delivery of concentrated drug thereby enhancing the drug stability and bioavailability. The biodegradable material usage in synthesis prevents toxic side effects of the drug. (Jong D W H et al 2008)

Problem Identification

Though chemotherapy is a better treatment for different stages of cancer, there have been reports of many side effects. Also, there is a need for targeted drug delivery to cancer cells. Use of plant extracts as nano-particles can function as a better anti-cancer agent and will help to bring a breakthrough in the field of cancer treatment.

Literature review

Terminalia chebula belongs to the family Combretaceae. According to Ayurveda, the herbal extract from fruit can eliminate waste from body and promotes tissue growth. It has been used as a formulation for antibacterial, antifungal, anti-inflammatory and anti-cancer activity. The major bio-active constituents of the fruit include tannins, chebulic acid , anthraquinones, chebulagic acid, chebulinic acid, , ellagic acid and gallic acid. The anticancer activity of the extract has been evaluated in Invivo and Invitro systems. It was tested on EAC cells in swiss albino mice and was noticed that the lifespan in mice was increased by 88% and Hb, RBC and WBC count was restored to normal values. (Ahuja R et al 2013). Studies on MCF-7, HOS-1, PC-3 and PNT1A cell lines have shown that it can decrease cell viability, inhibits cell proliferation, and induces cell death in a dose dependent manner . (Saleem A et al 2002) There are reports which states the apoptotic activity in lung cancer cell lines and also causes cell morphology changes. It acts by regulating the Bcl-2 family protein-mediated mitochondrial pathway. It increases the activation of caspase-3, induced PARP cleavage, and promotes cytochrome c release into the cytoplasm. (Wang M et al 2015)

Phyllanthus emblica also known as Indian Gooseberry or amla, has been used as a traditional medicine for different disorders. Anti-oxidant activity of the extract has been evaluated and cytotoxic activity against HT-29 cells lines has been evaluated. The viability of cells was significantly reduced. (Sumalatha D , 2013) Apoptosis activity of the extract in HeLa cells was observed which caused DNA fragmentation and increased activity of caspase-3/7 and caspase-8, and also up-regulation of the Fas protein was noted which could indicate a death receptor-mediated mechanism of apoptosis. (Ngamkitidechakul C et al 2010)

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Tinospora cordifolia belongs to the family of Menispermaceae. Tinospora extracts are widely used in the Ayurvedic system in treatment of various disorders. Studies have shown that the extract possess antimutagenic and anticancer activity. Apoptotic activity of the extract was evaluated on MCF7, MDA MB 231, and HeLa cancer cell line. Sub-Go phase was increased and S-phase diminished and caused inhibition of DNA replication. (Tungpradit R et al 2010) Studies have shown that the extracts had an effect in increased expression of pro-apoptotic gene, Bax, and decreased expression of anti-apoptotic gene, Bcl-2, in Ehrlich ascites tumor (EAT) in mice. (Thippeswamy G et al 2007)

Withania somnifera popularly known as Ashwagandha which belongs to the family of Solanaceae has therapeutic application due to presence pf natural antioxidants functioning as reducing agents, free radical scavengers. Efficient cytotoxicity was shown in MCF-7, PA-1 and A-459 cancer cell lines. (Nema R et al 2013) Invitro and Invivo asssays have shown that activation of tumor suppressor proteins p53 and pRB, hypophosphorylation of pRB, decrease in cyclin B1 and increase in cyclin D1. (Wadhwa R et al 2013)

Terminalia arjuna also known as kumbuk, belongs to the family of Combretaceae, has a history of medicinal uses in cancer treatment. Studies of the extract on Human hepatoma cells showed inhibition of the proliferation of HepG2 cells and apoptotic morphology was observed. DNA fragmentation, accumulation of p53 and cleavage of procaspase-3 protein were seen. (Sivalokanathan S et al 2006)

Asparagus racemosus is a native medicinal plant of the family Liliaceae. Anti tumor activity has been well studied in UOK146 cell lines and down regulation of PRCC-TFE3 fusion transcript was observed. (Verma P S et al 2014) There are studies which demonstrate apoptosis in HepG2 cells. The bioactivity was observed by binding to cell-surface receptors which caused increase in ROS levels (Ji Y et al 2012)

The advancements in nanotherapeutics have given a breakthrough in development of cancer therapeutics. There are reports of many herbal extracts which are formulated as Nanoparticles acts as effective anticancer agents. Nanocapsulated P. senega caused cell death and apoptosis in lung cancer cell line A549 and increased expression of caspase-3, p53 mRNAs of A549 cells. (Paul S et al 2011) Nanoparticles synthesized from Phyllanthus emblica have an effective anti proliferative activity. ROS generation, apoptotic morphological changes, mitochondrial depolarization and DNA damage was observed in Hep2 cell line. (Rosarina F S et al 2013)

Nanotechnology aims in target specific delivery of drugs which enhances bioavailability. Studies have shown that herbal extracts have potential anticancer properties. Nano capsulated herbal extracts can enhance the therapeutic potential.

Hence, this study will focus on the nanoparticle synthesis from the formulation of the extracts of Terminalia chebula, Phyllanthus emblica, Tinospora cordifolia, Withania somnifera, Terminalia arjuna and Asparagus racemosus and evaluation of anticancer property of the synthesized nanoparticles.


  1. Study of pharmacokinetic properties of the herbal extracts using Insilico approach
  2. Synthesis of nano-particles and conjugation of the herbal extracts
  3. Study of anticancer properties of the herbal extract using Invitro approach
  4. Evaluation of the effect of herbal extract on apoptotic genes


  1. Assessment of Pharmacokinetics properties of the herbal compound using Insilico approach

The phytoligand present in extracts of Terminalia chebula, Phyllanthus emblica, Tinospora cordifolia, Withania somnifera, Terminalia arjuna and Asparagus racemosus have been determined using experimental methods. The structures will be retrieved from the structural database such as PDB, CHEMISPIDER and NCBI PUBCHEM databases. The pharmacokinetic features of the phytoligands will be analyzed using PREADMET tool. Human intestinal absorption, Invitro skin permeability, Invitro MDCK cell permeability prediction will be performed to check absorption. Predcition of plasma protein binding, blood-brain barrier penetration will be carried out to check distribution of the phytoligands. AMES test and Rodent carcinogenicity values will be assessed for toxicity studies. Drug likeness features will be assessed based on Lipinski rule of five, CMC-like rules, MDDR-like rules and WDI-like rules.

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2. Synthesis of nanoparticles from the herbal compound

Gold nanoparticles will be used for the conjugation of the herbal extract. Gold ion solution will be prepared and appropriate amount of herbal extract will be mixed. The formation of nanoparticles will be verified using by UV-vis absorption and FTIR.

3. Cell culture and cytotoxicity assay

The synthesized nanoparticles will be tested on the Ca Ski (cervical cancer), and U87 (glioblastoma) cancer cell lines to assess anticancer activity. MTT assay will be performed to assess cell viability. The distribution of cells in different cell cycles will be studied using flow cytometry. To check for apoptosis, Annexin-V-FLUOS and PI staining kit will be used. Total Antioxidant capacity assay will be performed to check for antioxidant properties. Western blot analysis will be performed to check for mitochondrial apoptotic proteins.

4. Gene expression analysis

Gene expression studies will be carried out using Real -time PCR. RNA will be extracted from the cell lysates of nanoparticle treated cell and untreated cells. Differentially expressed genes will be assessed. Further statistical tests will be employed for normalization and other statistical significance assessment.

Possible outcomes

  1. ADMET properties predicted for the compound will have favorable pharmacokinetic properties.
  2. Standardization of synthesis of nanoparticles from herbal extracts will be performed and Stable nanoparticles will be synthesized from the extract.
  3. Anticancer properties of the extract will be analyzed by Invitro approach.
  4. Differentially expressed genes will be assessed in microarray studies and statistical significance analysis will be carried out.
  5. Nano encapsulated extract will have an effective anticancer property and can help in cancer therapeutic interventions.


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