NanoViricides Inc. (NNVC)

[changes_iv]

[Published online Mar 2013...]

4.2 Microfluidic methods

Microfluidics, the science and technology of manipulating nanoliter volumes in microscale fluidic channels, has shown that several labour-intensive and time-consuming steps such as sample preparation, mixing, reactions, purification, separations, and detection could be performed on a single monolithic microfabricated device.238 In the last few years, applications of microfluidics have expanded from conventional chemical and biological analysis to other fields such as chemical reactions, biochemical assays, and cell handling.239 Two particularly important contributions have been the development of soft lithography in PDMS as a method for fabricating prototype devices, and the simple fabrication of pneumatically activated valves, rapid mixers and pumps on the basis of soft-lithographic procedures. This has resulted in the fabrication of prototype devices to test new ideas in approximately 2 days (from design to working device), whereas the same applications for silicon technology may take a month ormore for non-specialists to carry out.With respect to nanoparticles, the ability of microfluidic systems to mix reagents rapidly, provide homogenous reaction environments, continuously vary reaction conditions, enable rapid temperature control, and allow addition of reagents at precise time intervals—are some of the key features that have made microfluidic systems useful for the synthesis of NPs.240 Furthermore, synthesis carried out in microchannels allows for in-line characterization,241 feedback control,242 and high-throughput continuous synthesis,243 which potentially enables screening and optimization of libraries of nanoparticles with different properties.

Recently, technologies developed for the synthesis of polymeric NPs have demonstrated the tremendous potential for microfluidics to dramatically improve on current bulk synthesis methods. Polymeric NPs prepared by bulk synthesis tend to have variable physicochemical properties (size, surface composition, and drug loading) due to the inability to control the mixing of precursors.236 Further post-processing by extrusion, freeze–thaw, sonication, and/or high-pressure homogenization is often required. Using rapid mixing techniques in micro-channels such as hydrodynamic flow focusing, polymeric NPs exhibiting narrow size distributions compared to bulk synthesis have been prepared in a reproducible manner (Fig. 4).236, 237 In these systems size can be tuned by either varying the mixing time of precursors, which is achieved by varying the flow ratio of the precursor streams, by varying the molecular weight of the polymer, or by simply varying the concentration of the polymer in the organic solution. Remarkably, for polymeric NPs prepared through microfluidics, higher drug encapsulation without increase in NP size has been observed,236 which is highly desired for therapeutic NPs. Another method to prepare NPs takes advantage of the rapid mixing microenvironment that occurs in micro-droplets formed inside microfluidic channels.244 For instance, cross-linked alginate NPs were synthesized in a micro-channel using aqueous alginate droplets as templates, followed by the shrinkage of the drops. This method exhibited remarkable control over the NP properties, specifically size and size distribution. These are just a few examples showing the advantages of microfluidics for nanoparticle synthesis, and recent reviews discuss these concepts further.240, 245 Given the volume of research currently involving the microfluidic synthesis of NPs, it is expected that as more therapeutic NPs reach a clinical stage, the need for improved synthesis methods would also increase, at which point microfluidic technologies could likely become an important tool in their development of NPs.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3684255/

WEST HAVEN, Conn., Feb. 25, 2015 /PRNewswire
NanoViricides Files Quarterly Report for Period Ending 2014-12-31
...
In addition, the Company is performing process development and scale up studies on its FluCide drug candidate. FluCide is designed as a treatment for influenza in seriously ill hospitalized patients, a potentially fatal disease for which no satisfactory treatment exists. FluCide's excellent safety profile resulted in the need for a very large quantity of the drug for the GLP Safety/Toxicology studies required for an Investigational New Drug ("IND") filing. The multi-kilogram quantities of FluCide needed for these safety and toxicology studies will be produced in the new Shelton facility.
http://www.prnewswire.com/news-releases/nanoviricides-files-quarterly-report-for-period-ending-2014-12-31-300041185.html

NanoViricides Provides an Update on Its Progress over the Last Quarter

Influenza
We are currently working to increase the production of FluCide™, our anti-influenza drug candidate. FluCide has been designed for use in hospitalized patients with complicated influenza. FluCide was found to be extremely safe in preliminary toxicology studies in mice and rats. As a result of the extreme safety finding, it was estimated that about 2.5kg of drug substance would be needed for the complete large animal toxicology studies. These studies are needed for filing an Investigational New Drug Application (IND). In mice, no adverse events were observed even at doses as high as 480 mg/kg/d repeated on five days (a total of 2,400 mg/kg), when given intraperitoneally. Similar strong safety was also observed in the initial part of the formal toxicology study in rats. In rats, no adverse events were observed with doses as high as 300mg/kg/d given by rapid intravenous infusion, and repeated for 14 days (a total of 4,200 mg/kg). We are in the process of producing a total of 2.5kg of FluCide for the final large animal toxicology studies. Our toxicology studies are being performed by BioAnalytical Systems, Inc. (BASI) of Indiana (BASI).

We are now working to optimize all of the processes involved in the production of FluCide. Equipment needed for this task is being acquired, and is being installed by factory representatives as it arrives. Some items have lead times of 6 to 8 weeks for delivery. We are working as quickly as possible on setting up the production processes at our new state of the art c-GMP-capable manufacturing facility in Shelton, CT.

We are happy to announce that our Biological Characterization Group has now completely moved to our Shelton, CT campus. We are implementing a phased move to Shelton so that there is minimal impact on our continuing operations. We plan on continuing to use our West Haven facility to maximize R&D efforts on our large number of drug development programs.
http://www.reuters.com/article/2015/04/06/nanoviricides-updates-idUSnPn7fzXxL+85+PRN20150406

On Monday, April 27, 2015

NanoViricides Discusses the Strong Effectiveness of Its Anti-Herpes Drug Candidates in a Lethal Animal Model of Dermal Herpes Infection
...

In another news, the Company reports that the process of commissioning of its new facility in Shelton is on course. Our Bio-Analysis Group has already moved operations to the new facility. Large Scale Chemistry Group is completing the necessary modifications to the facility to enable large scale production processes. Various laboratory instruments are being installed by vendors under warranty programs for installation qualification and operational qualification.
http://www.prnewswire.com/news-releases/nanoviricides-files-quarterly-report-for-period-ending-2015-03-31-300084664.html

NanoViricides Files Quarterly Report for Period Ending 2015-03-31
...
Scale-up of the FluCide™ anti-influenza drug candidate is progressing well. We are continuing the CMC studies (see below) on FluCide production processes. These studies are necessary to enable further scale-up from the current multi-100g scale of production to kg-scale production of our nanoviricides drug candidates. The 1kg-scale production is being set up at our new Shelton, CT facility and headquarters.
...
The Company reports that the process of commissioning of its new facility in Shelton is on course. Our Bio-Analytics Group has already moved operations to the new facility. Large Scale Chemistry Group is completing the necessary modifications to the facility. We implemented a phased program for commissioning the new facility. This has enabled us to continue to progress in all of our existing programs, without interruptions.
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The Company believes that the drug approval process for a topical herpes treatment could be relatively rapid, based on the strong effectiveness results. The Company intends to meet with its FDA advisory consulting group, namely, Biologics Consulting Group, Inc., to chart out the path towards approval. In addition, the Company intends to engage a Contract Research Organization (CRO) for further development of a topical anti-herpes drug into the regulatory approval pathway for US FDA as well as internationally. The Company intends to study various indications including HSV-1 cold sores, HSV-2 genital lesions, Herpes keratitis (an eye disease), and Shingles, among other possibilities.

The market size for herpes virus treatments is in excess of $2 billion annually. The Company believes that a drug that is superior to existing therapies could result in significantly expanded market size, as has been demonstrated in the case of HIV, Hepatitis C and other diseases. An estimate of over $40 billion for an effective anti-herpes drug may not be excessive, considering the near-complete penetration of the various herpesviruses in human population, and the repetitive and debilitating nature of illnesses that they can cause.
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We are now progressing to a 1kg scale-up of FluCide, and enabling in-process control instrumentation. We need to make approximately 2.5Kg of our FluCide drug candidate for further Tox Package studies because of the excellent safety demonstrated by this drug candidate in safety and toxicology studies in both mouse and rat animal models. CMC stands for "Chemistry, Manufacture, and Controls," and relates to being able to make the drug substance and the drug product in a reproducible fashion, batch after batch. CMC programs for nanomedicines are relatively complex compared to those of small molecules. We have focused on developing scalable, reproducible processes from the very onset, which has helped us minimize the process development time.
http://www.prnewswire.com/news-releases/nanoviricides-files-quarterly-report-for-period-ending-2015-03-31-300084664.html

And if the nanoviricides(R) are bioequivalent, once you get the scale-up recipe for FluCide(TM), the recipe for other nanoviricides(R) won't be far off, or will they?
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Scaleup with Repeatable Results

Achieving results in the lab is a critical step, but for most commercial applications it is only valuable if the results can be reproduced in a production environment. Every model in the Microfluidics product line is engineered with the end result in mind. By utilizing a linear scaleup method of the fixed-geometry interaction chambers, customers using Microfluidizer processors in the lab are guaranteed to attain the same repeatable and predictable results when scaling up to production volumes.
http://www.microfluidicscorp.com/applications/biotechnology/52