Current Pharmaceutical Biotechnology - Volume 10, Issue 7, 2009

Chlorophytum borivilianum Santapau & Fernandes (Liliaceae) also known as ‘Safed Musli’ is a traditional rare Indian medicinal herb which has many therapeutic applications in Ayurvedic, Unani, Homeopathic and Allopathic systems of medicine. Its roots (tubers) are widely used for various therapeutic applications. It is used to cure physical illness and weakness, as an aphrodisiac agent and revitalizer, as general sex tonic, remedy for diabetes, arthritis and increasing body immunity, curative for natal and postnatal problems, for rheumatism and joint pains, increase lactation in feeding mothers, as antimicrobial, anti-inflammatory, antitumor agent, also used in diarrhea, dysentery, gonorrhea, leucorrhea etc. It has spermatogenic property and is found useful in curing impotency, now it is considered as an alternative ‘Viagra’. Its root contains steroidal and triterpenoidal saponins, sapogenins and fructans which act as therapeutic agents and play vital role in many therapeutic applications. It is a rich source of over 25 alkaloids, vitamins, proteins, carbohydrates, steroids, saponins, potassium, calcium, magnesium, phenol, resins, mucilage, and polysaccharides and also contains high quantity of simple sugars, mainly sucrose, glucose, fructose, galactose, mannose and xylose. The commercial exploitation of this plant and their secondary metabolites, germplasm conservation and in vitro production of secondary metabolites for quality control are some of the major prospects of this rare medicinal herb. The focus of the present review is to galvanize the potential of therapeutic and nutritive values of this herb and production of their secondary metabolites. The in vitro tuber induction, extraction, purification and characterization of saponins are also discussed in the present review.

Momordica balsamina, African pumpkin (Cucurbitaceae), is a tendril-bearing, wild climber containing wide spectrum of medicinal and nutritional values and has been used as a traditional folk medicine in many countries. The leaves, fruits, seeds, and bark of the plant contain resins, alkaloids, flavonoids, glycosides, steroids, terpenes, cardiac glycoside, saponins having various medicinal importance viz. anti-HIV, anti-plasmodial, shigellocidal, anti-diarrheal, antiseptic, anti-bacterial, anti-viral, anti-inflammatory, anti-microbial, hypoglycemic, antioxidant, analgesic and hepatoprotective properties. The therapeutic agent ‘Momordin’ is capable of inhibiting the growth of HIV and other viruses. The leaves are also important source of nutrients having 17 amino acids with adequate mineral composition like potassium, magnesium, phosphorus, calcium, sodium, zinc, manganese and iron. It also helps to combat the problem of micronutrient deficiencies in soil and high value of protein and fat with low fibre content. High potassium content is a good source for the management of hypertension and other cardiovascular conditions. This plant is being promoted as a protein supplement for cereal-based diets in poor rural communities. The commercial exploitation of this plant for biopharmaceuticals and neutraceuticals are some of the prospective future potential of this wild herb. This review discusses the potential of medicinal and nutritional importance of this wild herb for health care management.

Creatine plays a central role in energy metabolism and is synthesized in the liver, kidney and pancreas. In healthy patients, it is transported via the blood stream to the muscles, heart and brain with high and fluctuating energy demands by the molecule creatine transporter. Creatine, although naturally synthesized in the human body, can be ingested in the form of supplements and is commonly used by athletes. The purpose of this review was to assess the clinical applications of creatine supplementation on paediatrics. Creatine metabolism disorders have so far been described at the level of two synthetic steps, guanidinoacetate N-methyltransferase (GAMT) and arginine: glycine amidinotransferase (AGAT), and at the level of the creatine transporter 1(CrT1). GAMT and AGAT deficiency respond positively to substitutive treatment with creatine monohydrate whereas in CrT1 defect, it is not able to replenish creatine in the brain with oral creatine supplementation. There are also data concerning the short and long-term therapeutic benefit of creatine supplementation in children and adults with gyrate atrophy (a result of the inborn error of metabolism with ornithine deltaaminotransferase activity), muscular dystrophy (facioscapulohumeral dystrophy, Becker dystrophy, Duchenne dystrophy and sarcoglycan deficient limb girdle muscular dystrophy), McArdle's disease, Huntington's disease and mitochondriarelated diseases. Hypoxia and energy related brain pathologies (brain trauma, cerebral ischemia, prematurity) might benefit from Cr supplementation. This review covers also the basics of creatine metabolism and proposed mechanisms of action.

A protocol for induction and establishment of Agrobacterium rhizogenes mediated hairy root culture of Gossypium hirsutum was developed through infection with the A4 strain and co-cultivation on hormone-free semi-solid MS medium with B5 vitamins. It resulted in the emergence of hairy roots from the leaf explants, 21 days after infection. The transformation of hairy roots was established by PCR amplification of rol B and rol C genes of the Ri plasmid. All root lines expressed gossypol, although distinct inter-clonal quantitative variations were noticed. Five independent hairy root lines were studied for their growth kinetics as well as gossypol production. The yield potentials of one of them superseded others, as well as the non-transformed, in-vitro grown control roots. The content of gossypol in hairy roots reached a level of 2.43 mg/g DW. It was 4.5 times higher than in vitro and 1.47 times higher than in vivo grown roots of G. hirsutum. Selection of high gossypol producing hairy root lines of G. hirsutum can provide an alternative source for large-scale production of gossypol.

Trastuzumab and pertuzumab are monoclonal antibodies, used for inhibiting the ErbB2 receptor which is over expressed in breast and ovarian cancer. In this study, we identified that the most detrimental single point mutation is from tryptophan to cysteine at the residue position of 452 on ErbB2 receptor by using I-Mutant 2.0, SIFT and PolyPhen programs. The modeled mutant showed less stability than native ErbB2 protein based on both total energy of the mutant and stabilizing residues in the mutant protein. This is due to deviation between the mutant and native ErbB2 having the RMSD of about 2.83Å. Further, we found, pertuzumab showed a marginal higher binding affinity with ErbB2 receptor of native and mutant type with a binding free energy of -16.01 kcal/mole each as compared to trastuzumab, showing a binding free energy of -15.26 kcal/mole in ErbB2 of native type. On the contrary, trastuzumab showed a remarkably high binding affinity with ErbB2 receptor of mutant type having the binding free energy -24.40 kcal/mol. Moreover, the reason for high binding efficiency of trastuzumab with mutant ErbB2 is due to additional hydrogen bonding of amino acid Asn30 of trastuzumab with Asp596 and Glu598 of ErbB2 receptor of mutant type. Based on this work, we propose that pertuzumab could be the potential monoclonal antibody against ErbB2 for native type and trastuzumab could be the potential monoclonal antibody against ErbB2 of mutant type. Therefore combined administration of trastuzumab and pertuzumab could be a novel strategy for breast cancer treatment.

Immune stimulating complexes (ISCOMs) incorporating recombinant hepatitis B surface antigen (rHBsAg) were prepared for induction of humoral and cellular immunity by subcutaneous administration. Prepared ISCOMs were characterized for their size, shape, incorporation efficiency, zeta potential, antigen integrity, antigen conformation and immunogenicity by biophysical and immunological techniques including transmission electron microscopy (TEM), Dynamic light scattering (DLS), SDS-PAGE, fluorescence spectroscopy, in vitro potency test and in vivo humoral and cellular immune stimulatory efficacy in Balb/c mice. Prepared ISCOM particles show characteristic cage like morphology with average size of 44 ∼nm, polydispersity index 0.1, negative zeta potential (-21.7 mV) and antigen association efficiency - 39%. Tryptophan emission fluorescence and in vitro potency assay data suggest that association of rHBsAg with ISCOMs results in local electrostatic interactions, motional restriction of tryptophan residues of the protein resulting in reduction of anti-rHBsAg monoclonal antibodies binding affinity. Immunization with rHBsAg ISCOMs resulted in upregulation of specific cellular (IFN-γ and IL-2) as well as IgG response (IgG2a isotype biased) humoral response in Balb/c mice. Immune responses were significantly higher than those produced by of alum-adsorbed antigen (alum-rHBsAg) after (one booster) (p < 0.001). These data demonstrate that although the conformation of rHBsAg after incorporation into ISCOMs was moderately altered but due to strong adjuvant ability, rHBsAg ISCOMs were highly immunogenic as compared to marketed rHBsAg formulations by subcutaneous route of administration.