Recent Patents on Biotechnology - Volume 7, Issue 1, 2013

Diseases caused by protozoan parasites are still an important health problem. These parasites can cause a wide spectrum of diseases, some of which are severe and have high morbidity or mortality if untreated. Since they are still uncontrolled, it is important to find novel drug targets and develop new therapies to decrease their remarkable social and economic impact on human societies. In the past years, human HSP90 has become an interesting drug target that has led to a large number of investigations both at state organizations and pharmaceutical companies, followed by clinical trials. The finding that HSP90 has important biological roles in some protozoan parasites like Plasmodium spp, Toxoplasma gondii and trypanosomatids has allowed the expansion of the results obtained in human cancer to these infections. This review summarizes the latest important findings showing protozoan HSP90 as a drug target and presents three patents targeting T. gondii, P. falciparum and trypanosomatids HSP90.

Although many patents reported bioactive peptides with numerous demonstrated bioactivities and potential applications, there exist some limitations to the production of large quantities to satisfy the growing market demands. Indeed, considering that most functional peptides are present in complex matrices containing a large number of hydrolyzed protein fractions, their separation and purification are required. Some advances have been made in the use of conventional pressure-driven processes for the continuous production and separation of peptides, however, most of these patented technologies are not scalable and demonstrate a low selectivity when separating similar sized biomolecules. To improve the separation efficiency, the use of an external electric field during pressure-driven filtration was proposed and patented. However, whatever the claims, the pressure gradient brings about the accumulation of peptides at the nearby membrane surface and affects the membrane transport selectivity. To overcome these drawbacks, a recent patent proposed the simultaneous fractionation of acidic and basic peptides, using a conventional electrodialysis cell, in which some ion exchange membranes are replaced by ultrafiltration ones. The perspectives in the field of peptide separation will be the development of new membrane materials and new equipments such as microfluidic devices to improve selectivity and yield of production.

Proteins and peptides are the building blocks of human body and act as the arsenal to combat against the invading pathogenic organisms for treatment and management of diseases. Majority of such biomacromolecules are synthesized by the human body itself. However, entry of disease causing pathogens causes misleading in the synthesis of desired proteins for antibody formation. In such alarming situations, the delivery of requisite protein and peptide from external source helps in augmenting the body's immunity. The major drawbacks underlying poor biopharmaceutical performance of high molecular weight protein and peptide drugs are due to poor oral absorption, formulation stability, degradation in the gastric milieu, susceptible to presystemic metabolism. Numerous literature recounts the application of myriad drug delivery strategies for the effective delivery of protein and peptides viz. parentral, oral, transdermal, nasal, pulmonary, rectal, buccal and ocular drug delivery systems. There are many reviews on various delivery strategies for protein and peptide pharmaceuticals, but the present review article provides a bird's eye view on various novel drug delivery systems used for enhanced delivery of protein and peptide pharmaceuticals in the light of patent literature. Apart from this, the present manuscript endeavor provides idea on possible causes and major degradation pathways responsible for poor stability of protein and peptide drugs along with recent market instances on them utilizing novel drug delivery systems.

Although thermal sterilisation is a widely employed industrial process, little work is reported in the available literature including patents on the mathematical analysis and simulation of these processes. In the present work, software packages have been developed for computer aided optimum design of thermal sterilisation processes. Systems involving steam sparging, jacketed heating/cooling, helical coils submerged in agitated vessels and systems that employ external heat exchangers (double pipe, shell and tube and plate exchangers) have been considered. Both batch and continuous operations have been analysed and simulated. The dependence of del factor on system / operating parameters such as mass or volume of substrate to be sterilised per batch, speed of agitation, helix diameter, substrate to steam ratio, rate of substrate circulation through heat exchanger and that through holding tube have been analysed separately for each mode of sterilisation. Axial dispersion in the holding tube has also been adequately accounted for through an appropriately defined axial dispersion coefficient. The effect of exchanger characteristics/specifications on the system performance has also been analysed. The multiparameter computer aided design (CAD) software packages prepared are thus highly versatile in nature and they permit to make the most optimum choice of operating variables for the processes selected. The computed results have been compared with extensive data collected from a number of industries (distilleries, food processing and pharmaceutical industries) and pilot plants and satisfactory agreement has been observed between the two, thereby ascertaining the accuracy of the CAD softwares developed. No simplifying assumptions have been made during the analysis and the design of associated heating / cooling equipment has been performed utilising the most updated design correlations and computer softwares.

In this research it was attempted to overexpress the acidic subunit, from the 11S amaranth seed globulin termed amarantin, modified with antihypertensive peptides in Escherichia coli Rosetta (DE3) by manipulating some factors in batch fermenter such as growth medium composition, inducer (isopropyl β-D-thiogalactopyranoside [IPTG] or lactose), air flow, cultivation temperature, agitation speed and induction time. The possibility of using several minimal media and lactose as inducer to increase yields of the recombinant protein was investigated. Previous fermentations at flask level showed that two minimal culture media (A6 and A7) and 0.5% (w/v) lactose presented high yields of the engineered protein expression. Thus, the latter two media were tested at fermenter level, the lactose inducer, and different environmental conditions. Factors with significant effects were identified by Plackett-Burman design with center points and were adjusted at the level suggested and the yields of the recombinant protein were increased from 303.2 to 1,531 mg L-1 in A6 and from 363.4 to 1,681 mg L-1 in A7. Unlike some patents where the highest productivity was achieved at 24 h or afterwards, in this research the best productivity of the recombinant acidic subunit was attained at 4 and 6 h of induction using both media, respectively.

This paper presents the design and test of a dual-mode electric and magnetic biological stimulator (EM-Stim). The stimulator generates pulsing electric and magnetic fields at programmable rates and intensities. While electric and magnetic stimulators have been reported before, this is the first device that combines both modalities. The ability of the dual stimulation to target bone and muscle tissue simultaneously has the potential to improve the therapeutic treatment of osteoporosis and sarcopenia. The device is fully programmable, portable and easy to use, and can run from a battery or a power supply. The device can generate magnetic fields of up to 1.6 mT and output voltages of +/- 40 V. The EM-Stim accelerated myogenic differentiation of myoblasts into myotubes as evidenced by morphometric, gene expression, and protein content analyses. Currently, there are many patents concerned with the application of single electrical or magnetic stimulation, but none that combine both simultaneously. However, we applied for and obtained a provisional patent for new device to fully explore its therapeutic potential in pre-clinical models.