Category Archives: Nano Medicine

As Covid-19 vaccinations increase globally, life is getting back to normal. However, it's no longer the world we experienced before the pandemic. The long-term nature of this global crisis has changed customer needs and daily lifestyles. This will, in turn, change what I expect the world to look like in 2022 and beyond. Let's review the top 10 technology and business trends that we'll likely encounter in the coming year.

Momentum in Life Science Technologies

The life science industry comprises pharmaceuticals,biotechnology, environmental sciences, biomedicine, nutraceuticals, neuroscience, cell biology, and biophysics. Partially because ofincreased investments in mRNA vaccine technology and high-performance Covid-19 testing, 2021 has driven great innovation in life science technologies.

Examples of this innovation include the mRNA-based vaccines developed by Pfizer-BioNTech and by Moderna. Recent Covid drug developments by Merck have also pushed the life science industry forward rapidly.

Looking ahead, I expect we'll see strong movement in the several life science sectors, including advanced research; application of robotics technology; artificial intelligence (A.I.) tools; use of cloud technology; the improvement of drug testing speed; integration of genetic information; and usage of gene technology. I also foresee the development of personalized medicine through advancements in genetics technology. The startup 23andMe, for example, helps people access, understand, and benefit from the human genome.

Networking and Interconnectivity: Expansion of 5G-6G and Satellite-Based Internet Usage

As remote working becomes commonplace, internet reliability becomes more vital than ever. The internet of things (IoT) makes the internet a more integral part of our lives, and developments at all network levels will continue to drive research and push the internet economy forward.

According to Cisco's Visual Networking Indexforecast update for 2018, there will be 1.4 billion more people using the internet by 2022, compared with 3.4 billion users in 2017. This equates to almost 60 percentof the global population, assuming a population of eight billion by 2022. By then, internet users are expected to consume 4.8 zettabytes of data per year, 11 times the amount of IP traffic generated in 2012, with 437 exabytes.

While 5G may be in its infancy, we'll see an increased focus on 6G in 2022. China started research into 6G in 2018. By late 2020, the country had launched a satellite to test terahertz signal transmission. Huawei and ZTE were involved. The U.S. also started 6G research in 2018 with the Federal Communications Commission (FCC) opening higher frequency spectrum for experimental use. A Next G Alliance was started in 2020, with companies on board including Apple, AT&T, and Google. Korea, Japan, and some European countries have started looking into 6G seriously, and we expect to see more announcements in 2022.

High-Performance Computing Becomes Mainstream

Given the recent growth of big data-based research and analysis and cloud-based computing, I expect high-performance computing usage to surge in 2022. From drug discovery to cancer research to space exploration, high-performance computing will become increasingly important, and quantum computing technology needs to keep up with business demand.

We've already seen massive development in quantum computing space by players including Google, IBM, Microsoft, Amazon, and Alibaba. Startups such as Rigetti Computing, D-Wave Systems, ColdQuanta,1QBit, Zapata Computing, and QC Ware have exceeded the industry expectations in their technology and their growth.

I expect rapid progress in quantum computing during 2022, because the high demand for larger machine power is going to accelerate development in this sector.We'll likely see announcements of large qubit developments in the new year. Commercial quantum computing will soon be within our reach, and new breakthroughs will occur at an accelerated pace.

Continued Growth in Artificial Intelligence, Big Data Analytics, and Cloud Computing

A.I. has become an integral part of our lives. From improvements to personalization, ranking search results, recommending products, and understandingand controlling devices tobuilding better models of the environment to automatingindustry, we see the application of artificial intelligence everywhere as we look toward 2022.

Organizations are using predictive analytics to forecast future trends. According to a report published by Facts & Factors, the global predictive analytics market is growing at a compound average growth rate of around 24.5 percentand is expected to reach $22.1 billion by the end of 2026.

We will also see massive growth in cloud computing. By 2022, the cloud will be more entrenched and more computing workloads will run on the cloud. The internet of things will push this space in a bigger way, for sure. According to predictions from Gartner, global spending on cloud services is expected to reach over $482 billion in 2022, up from $314 billion in 2020.

Internet Security and Privacy Dominate

As the world becomes more digital, we're seeing an increased number of personal and organizational cyberattacks around the world. Several corporations have been the target of major cyberattacks in recent days, and businesses have become more vulnerable to destructive attacks, in part because ofincreased remote work giventhe pandemic.

During 2022, I expect that we'll see cyberattacks across sectors, and we will see the industry take a variety of counteractive measures. Defending against cyberattacks will include educating individuals so they can identify and avoid network assaults, thereby safeguarding their company's image.

Applying A.I. will make robust cybersecurity protocols easier andless expensive to createand more effective than ever. Use of A.I. can enable cybersecurity to spot risks and learn the behaviors of cybercriminals, thus preventing prevent future attacks. It can also help cut down on the time it takes cybersecurity specialists to do their daily tasks.

The MetaverseWill Shine in 2022

The metaverse is a digital reality combining social media, online gaming, augmented reality, virtual reality, and cryptocurrencies to allow virtual user interaction. The metaverse may refer to anything on the internet, including all of AR's offerings. Virtual space of the future describes the metaverse, providing access to all kinds of entertainment and projects, including the opportunity to work.

We expect the metaverse to be immersive, ubiquitous, and free to access. It will be digitally focused, and potentially involves entertainment, social connection, work productivity, and behavior modification at scale. It will create an entire ecosystem for developers, apps, ads, and new digital innovations. The metaverse will facilitate convenience, consumption, and a frictionless access to services.

Only a few companies today have a real stake in the metaverse. They include ByteDance, Tencent, Facebook, Spotify, Zoom, Amazon (Twitch), Alibaba, Roblox, Bilibili, Snapchat, Kuaishou, and Huawei. Apple's wearables also point to the metaverse with AR glasses coming soon. Microsoft with Microsoft Teams and gaming is perhaps the most advanced company in the world regarding the metaverse. WeChat's ability to facilitate payments and access to a variety of services is the start of the metaversesuper apps.

As the metaverse ecosystem grows in 2022, I expect it will have a direct impact on the future of many technology sectors. These include gaming, wearables, VR and AR, collaborative productivity (Canva, Slack), consumer tech products (including AR glasses or smart speakers), social networks, educational technology, health technology, communication technology (Zoom), digital currencies, and convenience mobilization and on-demand consumption (Amazon, Meituan, Alibaba, JD.com, Shopify).

NFT Platforms Will Boom

A non-fungible token (NFT) is a digital object: computer code and data that conveys ownership of something. The property may be online:for example, virtual real estate in the digital world or special equipment in a video game. Or it could be real: real estate, a painting, or a seat at a concert. An NFT can also be a hybrid:for example, the right to decide who can rent a room in a cooperative living space. A baseball card, Ronaldo's first ball, a vintage car, or a piece of land in Central London: All are one of a kind and can be transferred to the NFT.

In 2022, we are likely to see NFTs everywhere; this includes in movies, TV shows, books, and more. NFTs are part of the digital economy and are becoming mainstream because they allow people to own something that represents a part of something bigger than themselves -- a piece of art or a character, for example. The digital economy is made up of a variety of online markets, including game economies, virtual real estate, and even social media platforms like Facebook.

Our society has changed rapidly over the past decade, especially because of the 2008 to 2009 economic crisis and the pandemic. These trends are coming together to shape a world where NFTs will beat traditional collectibles, making 2022 a tremendous year.

The Robotics Sector Will Become More Prevalent

We expect to see an increased use of robotics in everyday life during 2022. Usage will include the health care, agriculture, automotive, warehousing, and supply chain management sectors, and we will see more robotics-based automation that will continue to progress.

The Covid-19 pandemic presents both problems and opportunities forroboticscompanies in the logistics and supermarket industries. Unexpected strains on supply systems and product shortages have highlighted the need for better supply chain efficiency. It's also become obvious thatrobotsand automation providea safe means for manufacturers to keep employees socially separated yet allow the business to continue operating.

The use of robotics process automation (RPA) helps automate activities that are performed repeatedly; we expect RPA to grow in 2022 and become a standard technology for business. CIO reports that chief information officers are using RPA to free workers from boring, repetitive tasks and allow them to do more fulfilling work. Finally, we're seeing a rise in nanorobotics, tiny sensors with limited processing power. The first useful applications of these nano-machines may be in nanomedicine. Biological machines, for example, are used to identify and destroy cancer cells or deliver drugs.

Increased Urgency in the Renewable Energy Technology Sector

Climate change and dramatic news headlines are driving rapid growth in sustainable energy adoption. During the pandemic, this has been the only energy sector that has grown. According to the International Energy Agency (IEA), 40 percentmore green energy was generated and used in 2020; the agency expects continued growth in 2022. The falling costof generating sustainable energy also helps ensure that mass adoption is likely. While newer energy technologies like nuclear fusion, biofuel, and liquid hydrogen might take longer to come full circle, we look forward to serious advancements in 2022.

I also expect to see massive growth in the production and usage of solar and wind technology. Storage (battery) technologies are playing a larger role, since there will be an increased need to store renewable energy.

Blockchain Technology Prevails

Blockchain, an innovative breed of distributed ledger, enables companies to track a transaction and do business with unverified parties -- even without the assistance of financial institutions. This new capability significantly lessens business conflicts; it also adds other benefits such as append-only data structure, transparency, security, immutability, and decentralization.

Blockchain technology has enjoyed massive adoption and continues to disrupt many industries, from gaming and governance to finance. According to the International Data Corporation, companies will spend almost $6.6 billion on blockchain solutions in 2021, a 50 percentincreasefrom the previous year. This number is expected to grow beyond $15 billion in 2024. With the new rise of NFTs and themetaverse, blockchain will become more important in 2022.

Businesses are encouraged to start studying blockchain technology, since it is expected to generate significant business opportunities; Statista reports that global blockchain technology is predicted to grow to more than $23.3 billion by 2023. A notable existing blockchain deployment is that of Walmart's supply chain database application, allowing the monitoring of individual products directly to their very source, effectively elevating food supply standards. Amazon is rolling out its blockchain initiatives for 2019, which allow its Amazon Web Services clients to benefit from distributed ledger solutions.

Looking Ahead

There's little doubt that 2022 will see rapid progress in these and other technologies, as the world emerges from the Covid-19 pandemic. I see a prime opportunity for smart startups, corporations, and investors to capitalize on these emerging technology and business trends to become more successful than ever. Let's hope that the resulting innovation makes the world better for people and businesses.

The opinions expressed here by Inc.com columnists are their own, not those of Inc.com.

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The Top 10 Technology and Business Trends of 2022 - Inc.

Published: Oct. 5, 2021 at 8:00 AM EDT|Updated: 10 hours ago

BOSTON, Oct. 5, 2021 /PRNewswire/ -- NanoView Biosciences today announced the release of the ExoView R200, its next-generation platform for the sensitive detection and characterization of extracellular vesicles (EVs), including exosomes and viruses.

Launched in 2019, ExoView provides high-resolution sizing, counting and phenotyping of exosomes and viral vectors at the individual extracellular vesicle level. Understanding the biomarkers carried by extracellular vesicles has potential for diagnostic, prognostic, and therapeutic use for a broad range of diseases.

The original ExoView R100platform revolutionized EV detection, delivering EV sizing down to 50 nm, with high sensitivity and specificity that cannot be matched by existing EV characterization. The ExoView platform also requires low sample input, no extensive sample preparation or purification, and minimal hands-on time.

NanoView Biosciences is now taking the ExoView platform one step further with the release of the ExoView R200. In addition to the capabilities of the ExoView R100, the R200 new features include:

"We are very excited about the release of the R200 and the benefits it will provide to researchers in the EV field", said Jerry Williamson, CEO of NanoView Biosciences, "Based on the tremendous success of the ExoView platform, we have been working closely with our customers and scientific advisory boardto see what more is needed to address critical questions about EVs. We believe that the additional capability of the R200 will advance our goal to better understand the biological role of extracellular vesicles and their potential use as biomarkers for personalized medicine."

The ExoView R200 is available now, including upgrade paths for existing R100 users. For more information visit http://www.nanoviewbio.com.

About NanoView BiosciencesNanoView Biosciences, a Boston-based, privately-held company, is focused on enabling worldwide life science researchers to better understand the biological role of extracellular vesicles, including exosomes and viral vectors, and their potential use as biomarkers for improving the diagnosis, prognosis, treatment, and monitoring of disease. The Company's proprietary products, including the ExoView R100 and R200 platforms, have been designed to fully characterize exosomes and other extracellular vesicles for use in research and in the implementation of precision nanomedicine. ExoView is a high-throughput, cost-effective analysis platform that is easy to use and does not require purification or large sample volumes to accurately analyze exosomes.

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SOURCE NanoView Biosciences

The above press release was provided courtesy of PRNewswire. The views, opinions and statements in the press release are not endorsed by Gray Media Group nor do they necessarily state or reflect those of Gray Media Group, Inc.

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NanoView Biosciences Announces the Release of the ExoView R200 Platform - WWNY

Introduction

Pancreatic adenocarcinomas (PACs) are relatively rare compared to other cancers and represent only 3.2% of all new cancer cases in the US. Nevertheless, the average 5-year survival rate for all stages of PAC is only 10.0% because PACs cannot be detected and treated early. In general, the percent of cases and 5-year relative survival according to various stages of PAC (localized, regional, distant, and unknown) at diagnosis were reported to be 11% and 39.4%, 30% and 13.3%, 52% and 2.9%, and 7% and 6.1%, respectively, showing that the earlier PAC is caught, the better chance a person has of surviving 5 years after being diagnosed.1 Gemcitabine (GEM) is the only approved first-line monotherapy for treating PACs. Unfortunately, it still delivers unsatisfactory therapeutic outcomes in prolonging progression-free survival (PFS) and overall survival (OS) of patients with locally advanced and metastatic PAC.2

Combined therapy has become a major means to combat cancer thanks to its primary advantages of increased efficacy without, or with minimal, addictive toxicities at equal or reduced administered doses. Multiple combination therapies composed of GEM and different cytotoxic and biologic agents have undergone clinical evaluations to examine the therapeutic efficacies for patients at various stages of PAC since 2002 as reported by Lei et al.3 Among them, only the combination regimen of GEM with either erlotinib (Tarceva) or paclitaxel albumin-bound nanoparticles (NPs) (Abraxane) demonstrated significant improvements in most clinical outcome parameters compared to GEM alone, leading to the approval of both combination therapies by the US Food and Drug Administration (FDA) as a first-line combination therapy for patients with locally advanced and metastatic PAC in 20154 and 2013,5 respectively. Despite combination therapies having demonstrated improved outcomes in patient survival and quality of life, the overall improvement is still marginal, especially for patients diagnosed with late stages of the disease. There is still an urgent need to generate effective strategies, new single agents, or new combinations, to significantly improve clinical outcomes for treating PACs.

Several studies support the proteasome being an effective therapeutic target against PAC from the perspectives of high heterogeneity and chemoresistance. By unraveling transcriptomic predictive signature data by Fraunhoffer et al,6 a subgroup of PACs sensitive to FDA-approved carfilzomib (CFZ) was identified, and it was ultimately suggested to repurpose CFZ for treating PACs.7 Furthermore, proteasome inhibitors (PIs), such as carfilzomib, can induce apoptosis in PACs by inducing endoplasmic reticular (ER) stress, which facilitates synergistic effects when combined with radiation therapy or chemodrugs like camptothecin and paclitaxel.8 It was further disclosed that a combination of MG-132 (a PI) and camptothecin at a ratio of 5:1 (2.5 mol/l MG-132: 0.5 mol/l camptothecin) provided promising results with enhanced cytotoxicity compared to the single compounds in MIA PaCa-2 cells, while that for the combination of MG-132 and paclitaxel at the same 5:1 ratio but with lower concentrations of 0.08 mol/l MG-132 and 0.016 mol/l paclitaxel could moderately increase the cytotoxicity to 62% from 46% for paclitaxel alone at the same concentration of 0.016 mol/l as that in combination.9 This potentially suggests that a combination of PIs, such as CFZ with paclitaxel for treating PACs might be worth pursuing.

The poor biostability and short half-life of CFZ are considered major issues causing CFZ to perform with low efficacy in patients with solid cancers because it is difficult for CFZ to arrive at the proteasome in solid tumors.10,11 Polymer micelles (PMs) composed of biodegradable block copolymers poly(ethylene glycol) (PEG) and poly(caprolactone) (PCL) were reported to improve the metabolic stability of CFZ in vitro. However, despite the in vitro metabolic protection of CFZ, CFZ-loaded PMs or PEG-PCL-deoxycholic acid (CFZ-PMs) did not display superior in vivo anticancer efficacy in mice bearing human lung cancer xenografts (H460) to that of the clinically used cyclodextrin-based CFZ (CFZ-CD) formulation.12 A novel albumin-coated nanocrystal formulation of CFZ (CFZ-alb NC) displayed improved metabolic stability and enhanced cellular interactions, uptake, and cytotoxic effects in breast cancer cells in vitro. Consistently, CFZ-alb NCs showed greater anticancer efficacy in a murine 4T1 orthotopic breast cancer model than the currently used cyclodextrin-based formulation. It was highly suggested that human serum albumin (HSA)-bound NPs could be used as a viable nanocarrier to encapsulate CFZ for cancer therapy.13 As described above, it was paclitaxel (PTX) albumin-bound nanoparticles (Abraxane), not Taxol (a traditional dosage form with PTX being dissolved in the mixture of Cremophor EL and ethanol), which was approved for combination with GEM by the FDA as a first-line combination therapy for patients with locally advanced and metastatic PACs. Therefore, HSA-bound NPs could be used as a viable nanocarrier to co-encapsulate PTX and CFZ for combination therapy of PAC.

HSA has garnered considerable interest as a nanocarrier, due to its low toxicity, biocompatibility, and the ability to reduce interactions with phagocytes in the reticuloendothelial system (RES).1416 Moreover, albumin can interact with cancer cells based on its increased use as an energy source in rapidly proliferating cancer cells.17 It was reported that nanoalbumin-bound (nab)-drugs can aid drug permeation across tumor vessels.18,19 It was also suggested that albumin facilitates the movement of nab-drugs across endothelial cell membranes by binding to the gp60 receptor and sequentially interacting with other albumin-binding proteins such as secreted protein acidic and rich in cysteine (SPARC), which is abundantly expressed in and near cancer cells.2022 As exemplified, GEM (Gemcitabine)-loaded HSA and PTX-loaded HSA for practical PAC treatment have been reported by Han et al and Yu et al, respectively.23,24 Therefore, it was thought that NPs fabricated with HSA might potentially be an optimal choice for co-delivery of chemotherapeutic drugs with a high drug loading capacity, biodegradability, and good biocompatibility.

Abraxane is the first FDA-approved chemotherapeutic formulation based on Nab nanotechnology, which relies heavily on the use of organic solvents, namely, chloroform.25 The toxicity introduced by residual chloroform poses a potential risk to patient health. In response to the issue of chronic toxicity, a reversible self-assembling method, which eliminates the dependence on toxic organic solvents during manufacturing, was developed in a preliminary study and demonstrated to be capable of successfully preparing HSA-bound PTX and CFZ nanosuspensions. Furthermore, both NPs formed using this method still retained their suitability for intravenous (IV) administration.25 Therefore, in this study, the preparation of CFZ-loaded, PTX-loaded, and CFZ/PTX co-loaded HSA NPs was developed and optimized. To confirm these advantages, the properties of the three drug-loaded HSA NPs, including the encapsulating efficiency (EE), drug-loading (DL), mean size, polydispersity index (PDI), drug release, and cell growth inhibition against MIA CaPa-2 cells (human pancreatic cancer cell line) were characterized in vitro. Furthermore, the in vivo pharmacokinetic study of the three drug-loaded HSA NPs (CFZ/HSA NPs, PTX/HSA NPs, and CFZ/PTX/HSA NPs) were evaluated in Sprague-Dawley rats and compared to two solvent-based (Sb) drugs of CFZ and PTX (Sb-CFZ and Sb-PTX). The anti-tumor efficacy and systemic toxicity were further evaluated in MIA CaPa-2 tumor-bearing C.B-17 SCID mice.

Sb-CFZ (solvent-based CFZ) was prepared by dissolving 60 mg of CFZ (Chunghwa Chemical Synthesis & Biotech, New Taipei City, Taiwan), 3000 mg sulfobutylether beta-cyclodextrin (SBE--CD), and 57.7 mg citric acid in 29 mL deionized water through sodium hydroxide (NaOH) pH adjustment (pH=3.5). The solution was lyophilized and stored at 4C until reconstitution for use.26 Sb-PTX (solvent-based PTX) was prepared by solubilizing 6 mg PTX (ScinoPharm, Tainan, Taiwan) in 527 mg of purified Cremophor EL (polyoxyethylated castor oil; BASF, Ludwigshafen, Germany) in 497 mg (v/v) of dehydrated alcohol.

Drug-loaded HSA NPs were prepared with defatted human serum albumin (HSA) by a self-assembling method developed in our lab. Defatted HSA was produced by adsorption of fatty acids in HSA onto charcoal as previously described.27 Briefly, a marketed 20% HSA solution (Taiwan Blood Services Foundation, Taipei, Taiwan) was diluted with deionized water, and then the pH was adjusted to 2.7 with 1 N HCl. After adding 5 g of activated charcoal, the resulting HSA solution was stirred at 300 rpm and 4C for 2 h. The mixed solution was centrifuged at 8000 rpm and 4C for 10 min, and the supernatant was filtered through a 0.45-m nylon membrane (ChromTech, Bad Camberg, Germany) to remove the charcoal. Finally, the pH of the filtrate was adjusted to 7.0 with 1 N NaOH and lyophilized. The so-obtained lyophilized HSA powder was stored at 4C.28

The preparation of drug-loaded HSA NPs was divided into three steps. First, the pH of the HSA solution was adjusted to 2.7 with 1.0 N HCl to expose the hydrophobic domains.27,28 Second, the targeted hydrophobic drug in ethanol was added followed by stirring for 5 min to enhance interactions between the drug and HSA. Finally, the pH value was re-adjusted to neutral with 0.1 N NaOH to induce self-assembling and encapsulate the hydrophobic drug. Then, used high-pressure homogenization with an N2-3D Nanolyzer (Gogene, Hsinchu, Taiwan) to form stabilized drug-loaded HSA NPs. An Amicon Ultra-15 centrifugal filter (with a molecular weight (MW) cutoff of 10 kDa) was used to remove the ethanol, salt, and free drug, and then the drug-loaded HSA NPs were concentrated. The drug-loaded HSA NP concentrate was passed through 0.20-m regenerated cellulose filtration (Phenomenex, Torrance, CA, USA) to obtain translucent dispersion with typical diameter around 150 nm. Finally, lyophilized the solution for 48 hours without cryoprotectant.

The formulation and optimal homogenizer parameters utilized in step 3 for preparing CFZ-loaded HSA NPs (CFZ/HSA NP), PTX-loaded HSA NPs (PTX/HSA NP), and CFZ/PTX-loaded HSA NPs (CFZ/PTX/HSA NP) are described below. To prepare CFZ/HSA NPs, 200 mg CFZ was dissolved in 20 mL absolute alcohol, and the dispersion was added to 200 mL 0.9% defatted HSA solution. The ratio of drug to HSA was 1:9, and a 10K psi homogenizer parameter was applied for 10 cycles. To prepare PTX/HSA NPs, 300 mg PTX was dissolved in 12 mL absolute alcohol, and the dispersion was added to 120 mL 2.25% defatted HSA solution. The ratio of drug to HSA was also 1:9, and the 20K psi homogenizer parameter was applied for 20 cycles. To prepare CFZ/PTX/HSA NPs, 60 mg CFZ and 120 mg PTX were dissolved in 18 mL absolute alcohol, and the dispersion was added to 180 mL 1% defatted HSA solution. The ratio of both drugs to HSA was 1:10, and the 10K psi homogenizer parameter was applied for 10 cycles.

The mean particle size, size distribution, zeta potential, and polydispersity index (PDI) of drug-loaded HSA NPs were measured with a Zetasizer nano ZS (Malvern, Worcestershire, UK) by scattering angle of 90 at 25C. The drug-loaded HSA NPs were diluted with double-distilled water before the measurement, and all measurements were performed at least in triplicate. The shape and size were also observed by transmission electron microscopy (TEM), using Hitachi H-7000 (Hitachi, Tokyo, Japan). The purified NPs were diluted with water to allow clearer pictures to be taken. Samples were prepared by placing a drop on carbon-coated copper grids and sponging off the excess with filter paper. Then, the samples were stained with uranyl acetate (2% aqueous solution) for 3 minutes and dried at room temperature.28

To assess the entrapment efficiency of CFZ or PTX in drug-loaded HSA NPs, 10 mg lyophilized NPs was dissolved in 1 mL deionized water; then 9 mL acetonitrile was added and vortexed it for 1 minute. The solution was centrifuged at 14,000 rpm for 10 minutes. After appropriate dilution, CFZ or PTX in the supernatant was directly quantified by Waters alliance HPLC (Waters, Milford MA, USA) equipped with an Inert Sustain C18 column (150 4.6 mm, particle size 5 m, GL Sciences, Tokyo, Japan). The mobile phase was composed of acetonitrile and 0.05% formic acid aqueous solution (50:50, v/v, at a flow rate of 1 mL/min). The total analytical time for a single injection was 12 min. The injection volume was 10 L, and chose 210-nm wavelength for detection. The column oven was kept at 35C, and the sample cooler was maintained at 10C. The drug loading (DL) and entrapment efficiency (EE) of nanoparticles were calculated by the following equations:

and

;

WM is the weight of the drugs in the NPs, WI is the weight of the initial feeding drug, and WP is the weight of the initial feeding HSA.

Cell viabilities of the CFZfree (CFZ dissolved in DMSO), Sb-CFZ, CFZ/HSA NPs, PTXfree (PTX dissolved in DMSO), Sb-PTX, PTX/HSA NPs, and CFZ/PTX/HSA NPs were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for MIA PaCa-2 cell line obtained from ATCC. Cells were seeded at a density of 3 103 cells/well in 96-well plates and incubated for 24 h at 37C with 5% CO2. Then, tumor cells were treated with different concentrations (0.01, 0.1, 1, 5, 10, 50, 100, and 1000 ng/mL) of CFZfree, Sb-CFZ, PTXfree, Sb-PTX, or drug-loaded HSA NPs. After incubation for 72 hours, 200 L MTT (0.5 mg/mL) was added to each well for 2 hours. After removing the medium, 50 L DMSO was added to each well and gently shaken to dissolve any purple formazan crystal. The absorbance was measured at 550 nm (Bio-Tek). The survival rate was calculated using the following formula: percentage (%) cell survival = [(mean absorbency in test wells)/(mean absorbency in control wells)] 100. Values of the combination index (CI) were calculated by the Chou-Talalay method:29,30

C and P denote IC50 values of CFZ and PTX in combination therapy that inhibits 50% of the cell. C50 and P50 denote doses of CFZ and PTX that inhibit 50% cells alone. Values of CI = 1, CI < 1, and CI > 1, respectively, indicate additivity, synergy and antagonism.

Drugs released from the formulations were investigated in PBS (containing 0.5% Tween 80) by the dialysis method. The CFZ- or PTX-loaded HSA NPs were diluted to 0.1 or 0.2 mg/mL in 1-mL solution and then placed in a dialysis bag (OrDial D80-MWCO 60008000, cat. no. 60082530, Orange Scientific, Braine-lAlleud, Belgium) against 40 mL release medium, with 100 rpm shaking speed at 37C. Sampled 1 mL at 1, 2, 3, 4, 6, 8, 12, 24, and 48 hours, and performed the analysis using the HPLC method above.31

Male BALB/c mice (BioLasco Taiwan, Yilan, Taiwan) at 7 weeks of age were randomized into 4 groups and each group contained 4 mice. For single-dose study, we used tail vein injection at 0 days. For multi-dose study, we repeated half dose of single-dose study at 0 and 1 day. If there was no obvious toxic reaction, the dosage was elevated correspondingly. The weight changes and physiological signs were observed and recorded for 5 consecutive days in the first week. During the second week, the related assessment would be performed every 2 days. The whole study continued for 15 days. It would be specified as the maximum tolerance dose if there is any event for neurotoxicity, weight loss >20% or death.

Male Sprague-Dawley rats (BioLasco Taiwan, Yilan, Taiwan) at 8~10 weeks of age were used to study pharmacokinetic profiles after administration of Sb-CFZ, Sb-PTX, CFZ/HSA NPs, PTX/HSA NPs, Sb-CFZ+Sb-PTX, CFZ/HSA NPs+PTX/HSA NPs, and CFZ/PTX/HSA NPs. Rats were given a single tail vein injection of 5 mg/kg CFZ and 10 mg/kg PTX for each formulation (three or four rats per group). Blood samples were collected from the jugular vein in heparinized tubes at 0.017, 0.033, 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24, and 48 h after administration. All blood samples were immediately centrifuged at 4500 rpm for 10 minutes to obtain plasma, and then stored at 80C until analyzed by UPLC interfaced with MS (Triple Quadrupole Mass Spectrometry, TQ-XS, Waters). CFZ and PTX were extracted from the plasma as follows: 100L plasma was extracted with tert-butyl methyl ether (400 L) containing an internal standard (500 ng/mL chlorpropamide (Sigma-Aldrich, St. Louis, MO, USA) and 500 ng/mL docetaxel (ScinoPharm Taiwan, Tainan, Taiwan)) by vortex-mixing for 1 min. After centrifugation at 14,000 rpm for 10 min, 300 L organic phase was transferred to a new tube and dried at 40C. Samples were reconstituted in 100 L mobile phase and transferred to a new vial for the UPLC-MS/MS analysis. The measurement by UPLC-MS/MS. Chromatographic separation was performed with a Purospher Star RP-18 end-capped column (2.1 50 mm, particle size 2 m, Merck) and gradient elution (at a flow rate of 0.3 mL/min). The mobile phase comprised 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The total analytical time for a single injection was 5 minutes, and the injection volume was 2 L. The column oven was kept at 40C, and the sample cooler was kept at 10C. Detection of ions was performed in the positive ionization mode with the following transitions in multiple reaction monitoring mode (MRM): 720.33 100.03 for CFZ, 277.06 174.87 for chlopropamide (the internal standard for CFZ),32 854.29 104.99 for PTX, and 830.40 549.24 for docetaxel (the internal standard for PTX). The capillary voltage was 3.0kV, cone voltage was 30V, desolvation temperature was 350C, desolvation gas flow was 650L/h, and collision gas flow was 25L/h.

In vivo pharmacokinetic parameters including the area under the plasma concentrationtime curve (AUC), the apparent volume of distribution (V), plasma clearance (CL), and elimination half-life (T1/2) of each formulation were calculated and expressed by the mean and standard deviation (SD). The AUC0-1h, AUC0-2h, AUC0-24h, and AUC0-infinity were estimated by linear trapezoidal method. Plasma clearance (CL) was calculated from the dose/AUC0-infinity. The initial half-life (T1/2,initial) and terminal half-life (T1/2,terminal) values were calculated as ln(2)/k, where k represents either the initial distribution rate constant or the terminal elimination rate constant obtained from the slope of a semilogarithmic plot of the concentrationtime profile. The volume distribution (V) was estimated using a noncompartmental method provided by WinNonlin software (vers. 6.3.0.395, Pharsight, Princeton, NJ, USA). The maximum plasma concentration (Cmax) was recorded as observed for the first sampling time point, and C0 was the concentration at t = 0 (extrapolated).

C.B-17 female SCID mice, at 6~7 weeks of age (BioLasco Taiwan), were used as the tumor xenograft models. The models were established by subcutaneously inoculating MIA PaCa-2 (2107 cells/mouse, 100-L injection) into the right dorsal flank of each mouse. MIA PaCa-2 tumor-bearing mice with 150 mm3 tumor volumes were randomly divided into eight treatment groups (n = 5). One group of mice received an intravenous injection of saline as a control. The other groups received an injection of Sb-CFZ, Sb-PTX, CFZ/HSA NPs, PTX/HSA NPs, Sb-CFZ+Sb-PTX, CFZ/HSA NPs+PTX/HSA NPs, or CFZ/PTX/HSA NPs (equivalent to 5 mg/kg CFZ and 10 mg/kg PTX for each mouse). Administration was performed on days 0, 1, 7, 8, 14, and 15. Body weights (BWs) and tumor sizes were measured three times every week using digital calipers, and tumor volumes (mm3) were calculated. After being sacrificed by CO2 on day 46, tumors were harvested and weighed. The tumor growth inhibition (TGI) (%) was calculated as follows:33

Female C.B-17 SCID mice, at 6~7 weeks of age (BioLasco Taiwan), were used as the tumor xenograft models. The models were established by subcutaneously inoculating MIA PaCa-2 cells (2107 cells/mouse, 100-L injection) in the right dorsal flank of each mouse. On day 14 after tumor cell inoculation when tumor volumes had reached about 150 mm3, each mouse was given Sb-CFZ, Sb-PTX, CFZ/HSA NPs, PTX/HSA NPs, Sb-CFZ+Sb-PTX, CFZ/HSA NPs+PTX/HSA NPs, or CFZ/PTX/HSA NPs (equivalent to 5 mg/kg CFZ and 10 mg/kg PTX in each mouse) by an intravenous injection. After 2 and 8 h, mice were sacrificed by anesthesia and perfused with a PBS solution to remove the blood. The heart, lungs, liver, spleen, kidneys, and tumors were excised, weighed, and stored at 80C. Tissues were homogenized by an ultrasonicator probe (VCX 750; Sonics & Materials, Newtown, CT, USA) with 5 W and three pulses for 10 s. After that, 400 L of a PBS/0.1% heparin solution was added. Tissue homogenates (200 L) were obtained, and drug concentrations were analyzed by UPLC/MS/MS.

Data are presented as the mean SD of three different replicates. For in vivo studies, a one-way analysis of variance (ANOVA) with Tukeys multiple comparisons was used to test for significant differences in the longitudinal tumor volume growth over the entire experimental period among the eight treatment groups and to determine whether there was a significant interaction effect between CFZ and PTX. Significant differences between groups were indicated by *p<0.05 and **p<0.005.

As depicted in Figure 1, the mean particle sizes of CFZ/HSA NPs, PTX/HSA NPs, and CFZ/PTX/HSA NPs were 114.50.6, 117.40.4, and 105.30.6 nm; PDI values were 0.1440.007, 0.1660.007, and 0.1670.008; zeta potentials were 23.000.70, 21.10.58, and 21.30.70 mV; entrapment efficiencies (EEs, %) were 95.62.1%, 97.13.5%, and 92.72.6%/90.73.1%; and drug loadings (DLs) were 9.40.1%, 10.10.2%, and 9.10.4%, respectively. In addition, all zeta potentials measured were between 21.1 and 23.00 mV and indicated that the drug was encapsulated with HSA, which possessed a negative charge at a neutral pH because its PI was equivalent to 4.7.34 Figure 1 demonstrates that the three different HSA NPs were spherical, and the mean particle sizes were <200 nm under 80,000 TEM observation. Another noteworthy result was that the entrapment efficiencies for CFZ and PTX were, respectively, observed to be 92.72.6% and 90.73.1% in CFZ/PTX/HSA NPs. The calculated ratio for PTX and CFZ was 1.96; in other words, the method was suitable for preparing HSA NPs by achieving designed ratio nearly 2. Also, the high entrapment efficiency and drug loading (9.1%) indicated that HSA could act as novel and excellent nanocarriers for co-loading two drugs. In addition to the good compatibility with the two hydrophobic drugs, the preparation is simple and efficient with no cryoprotectant required.

Figure 1 TEM image and particle size analysis of drug/human serum albumin (HSA) nanoparticles (NPs).

The MIA PaCa-2 cell line was used as a model in the cytotoxicity study. Table 1 shows the values of the 50% inhibitory concentration (IC50) for different combinations of free drug in DMSO (CFZfree and PTXfree), two solvent-based drugs (Sb-CFZ and Sb-PTX), and two drug-loaded HSA NPs (CFZ/HSA NPs and PTX/HSA NPs). Respective IC50 values for CFZfree, Sb-CFZ, CFZ/HSA NPs, PTXfree, Sb-PTX, and PTX/HSA NPs were 8.9, 8.12, 7.89, 0.44, 0.47, and 0.86 ng/mL. The results indicated that the two model drugs maintained similar cytotoxicities in the different formulations. Compared to CFZ, PTX demonstrated a higher cytotoxicity toward the MIA PaCa-2 cell line. Table 1 also reveals the synergism of CFZ and PTX at ratios of 1:2, 1:1, and 2:1. IC50 values for free CFZ and PTX ratios of 1:2, 1:1, and 2:1 were 0.25, 1.39, and 1.00 ng/mL, respectively. According to the equation of the Chou-Talalay method, synergisms existed with free CFZ and PTX ratios of 1:2 and 2:1, and CI50 values were 0.38 and 0.83. For the Sb-CFZ and Sb-PTX combination ratios of 1:2, 1:1, and 2:1, IC50 values were 0.03, 0.37, and 0.51 ng/mL, and CI50 values were 0.05, 0.42, and 0.40, respectively. These results also revealed the synergism of Sb-CFZ and Sb-PTX. As for CFZ and PTX HSA NP ratios of 1:2, 1:1, and 2:1, IC50 values were 0.01, 0.07, and 0.66 ng/mL, and CI50 values were 0.01, 0.04, and 0.31, respectively. Figure 2 illustrates that the original product and HSA combinations exhibited synergism in all ratios examined. Since the combination ratio of 1:2 (CFZ: PTX) demonstrated the more-obvious synergic effect with IC50 value of 0.1 ng/mL and CI50 value of 0.08, the co-encapsulated ratio of 1:2 for CFZ and PTX in HSA (CFZ/PTX/HSA NPs) was chosen as the target formulation for the following assessment studies including the drug release study.

Table 1 50% Inhibitory Concentration (IC50) Values of Different Combinations of Carfilzomib and Paclitaxel

Figure 2 Characterization of the synergistic activity of combined carfilzomib (CFZ) and paclitaxel (PTX) treatment at different weight ratios.

Release percentages of CFZ and PTX from various formulations were assessed, and results are shown in Figure 3A and B, respectively. As shown in Figure 3A, the release of CFZ from Sb-CFZ, CFZ/HSA NPs, CFZ/HSA NPs+PTX/HSA NPs, and CFZ/PTX/HSA NPs were observed to have reached a plateau at 12~24 h with similar profiles, and release percentages at 12 h were determined to be 68.6%7.1%, 79.4%4.9%, 71.2%7.9%, and 71.7%3.8%, respectively. Furthermore, over 90% of CFZ had been released from CFZ/HSA NPs and CFZ/HSA NPs+PTX/HSA NPs at 24 h. We observed that the more-rapid release of CFZ from Sb-CFZ might be attributed to the use of the hydrophilic SBE--CD solubilizer to increase the solubility of CFZ in water. Similarly, the more-complete release of CFZ from the two HSA formulations was probably due to both being encapsulated in HSA NPs, which are expected to have greater surface areas for release. However, the release of CFZ from Sb-CFZ+Sb-PTX was measured at only 31.5%7.2% at 12 and 56.3%8.8% at 48 h, which were slower than that of CFZ released from Sb-CFZ at 12 h. This indicates that the addition of Sb-PTX to Sb-CFZ might have retarded the release of CFZ from Sb-CFZ resulting in a smaller release percentage. We suspect that by mixing Sb-CFZ with Sb-PTX, CFZ was encapsulated within the hydrophobic interior of Cremophor micelles, which was used as a solubilizing agent in the Sb-PTX formulation, causing retardation of permeation across the membrane of the dialysis bag to release CFZ.35

Figure 3 Drug release profiles of carfilzomib (CFZ, A) and paclitaxel (PTX, B). *p<0.05 and **p<0.005.

As for the release of PTX revealed by Figure 3B, release percentages of PTX from PTX/HSA NPs, CFZ/HSA NPs+PTX/HSA NPs, and CFZ/PTX/HSA NPs were observed to have reached a plateau at 12 h with similar profiles, and release percentages at the plateau were, respectively, determined to be 73.2%15.3%, 82.2%0.6%, and 65.6%8.8%. However, the release percentages of PTX from Sb-PTX and Sb-CFZ+Sb-PTX followed a gradually increasing trend, but these forms were only able to release 27.3%6.8% and 18.3%6.9%, respectively, at 48 h. Since both Sb-PTX and Sb-CFZ+Sb-PTX contained Cremophor as the solubilizing agent for PTX, it was expected as described above that the release of PTX trapped in Cremophor micelles would be retarded resulting in a slower release rate being observed. Therefore, CFZ and PTX were released more completely from the drug-loaded HSA NPs since both were encapsulated in HSA NPs, which were expected to have greater surface areas for release. Similarly, the greater extents of release percentages of PTX from the three HSA formulations were probably due to all of them being absorbed onto HSA in HSA NPs, which were expected to present as amorphous form to have higher solubility for increasing the extent of release.

In addition, it is worth mentioning that CFZ/PTX ratios released from co-encapsulated HSA NPs (CFZ/PTX/HSA NPs) were about 1.77~2.08 after 8 h of dissolution, which were consistent with the ratio of CFZ/PTX loaded in HSA NPs. However, the ratios of release amounts between CFZ and PTX from Sb-CFZ+Sb-PTX and CFZ/HSA NPs+PTX/HSA NPs were around 1.06~1.18 and 1.63~2.92, respectively, which did not reach the designed optimal ratio of 1:2 for synergism. It was concluded that release from the CFZ/PTX/HSA NP formulation conformed to the design combination ratio of 1:2 for CFZ and PTX to establish a potential synergistic effect.

Maximum tolerance dose for drug-loaded HSA NP and two free drugs as sb-CFZ and sb-PTX was then evaluated in BALB/c mice. Major dose limiting toxicities of CFZ or PTX were determined by neurotoxicity, weight loss >20% or death. According to the result (Table 2), the established maximum tolerance doses for a single dose were: Sb-CFZ 5 mg/kg, CFZ/HSA 17.5 mg/kg, Sb-PTX 20 mg/kg, PTX/HSA 300 mg/kg, Sb-CFZ+Sb-PTX 3.75/7.5 mg/kg, CFZ/HSA+PTX/HSA 10/20 mg/kg and CFZ/PTX/HSA 10/20 mg/kg. For multi-dose were as follows: Sb-CFZ 2.5 mg/kg, CFZ/HSA 5 mg/kg, Sb-PTX 12.5 mg/kg, PTX/HSA 150 mg/kg, Sb-CFZ+Sb-PTX <2.5/5 mg/kg, CFZ/HSA+PTX/HSA 5/10 mg/kg and CFZ/PTX/HSA 5/10 mg/kg. It is demonstrated that the maximum tolerance dose for drug-loaded HSA NP is normally higher than that for solvent-based form of free drug. Compared with solution base form, the combination or co-load HSA NP has at least 2 times higher maximum tolerance dose. Dr. Ernsting reveals that the maximum tolerance dose of single-dose for Abraxane performed in BALB/c is 170 mg/kg. However, in this research, PTX/HSA shows the quite remarkable tolerability from maximum tolerance dose study (300 mg/kg).36 Moreover, Dr. He also found that HSA encapsulation could lower the systemic nervous toxicity from VM-26.37 Taking the advantages with loading multi-drug in HSA NPs, the goal of achieving higher efficacy with lower toxicity was accomplished with such a multi-drug HSA NPs technique platform.

Table 2 Maximum Tolerance Dose Study for Various Combination Ratios of Carfilzomib and Paclitaxel on BALB/c Mice (n = 4)

Drug concentrations in plasma after a single tail vein injection of two solvent-based drugs (Sb-CFZ and Sb-PTX) and three drug-loaded HSA NPs (CFZ/HSA NPs, PTX/HSA NPs, and CFZ/PTX/HSA NPs) with respective dosing amounts of CFZ and PTX equivalent to 5 and 10 mg/kg are demonstrated in Figure 4A for CFZ and Figure 4B for PTX. Calculated pharmacokinetic parameters are listed in Table 3 for CFZ and Table 4 for PTX. As shown in Table 3, there were no significant differences among AUC0-1h, AUC0-24h, and AUC0-infinity obtained for all the various formulations after a single IV bolus administration of CFZ equivalent to 5 mg/kg. This indicates that CFZ was rapidly distributed to tissues and was quickly cleared from the systemic circulation after IV administration, resulting in the most reliable measure of the drugs bioavailability AUC in a period of 0 to 1 h (AUC0-1h) representing nearly the entire extent of the dosing amount of CFZ entering the systemic circulation. It was reported that the in vivo potency of CFZ is determined by the total dose administered (AUC), not Cmax, since CFZ can be rapidly distributed to tissues after IV administration as demonstrated by the potent proteasome inhibition in a variety of tissues.26 Because of this, pharmacokinetic parameters of T1/2,initial (min) and AUC0-1h (hr*g/mL) which are potentially related to the in vivo potency of CFZ were selected for comparison. Results in Table 3 demonstrate that T1/2,initial (min) and AUC0-1h (hg/mL) for CFZ after administration of Sb-CFZ were shorter and lower, respectively, than those for administration of CFZ/HSA NPs (12.722.14 vs 15.191.38 min and 0.2220.034 vs 3.3374.306 hg/mL), while neither of them was much different from administration of Sb-CFZ+Sb-PTX (12.722.14 vs 11.028.98 min and 0.2220.034 vs 0.2410.056 hg/mL). On the other hand, T1/2,initial (min) and AUC0-1h (hg/mL) for CFZ after administration of CFZ/HSA NPs were longer and much higher, respectively, than those with administration of CFZ/HSA NPs+PTX/HSA NPs (15.191.38 vs 9.142.93 min and 3.3374.306 vs 0.0550.009 hg/mL), while they were longer and much higher, respectively, than those with administration of CFZ/PTX/HSA NPs (15.191.38 vs 10.004.08 min and 3.3374.306 vs 0.5370.451 hg/mL).

Table 3 Pharmacokinetic Parameters of Carfilzomib Obtained from a Single Intravenous Bolus Administration of Various Formulations (Equivalent to 5 mg/kg Carfilzomib)

Table 4 Pharmacokinetic Parameters of Paclitaxel Obtained from a Single Intravenous Bolus Administration of Various Formulations (Equivalent to 10 mg/kg Paclitaxel)

Figure 4 Plasma concentrationtime curves of carfilzomib (CFZ, A) and paclitaxel (PTX, B) after intravenous administration at respective doses of 5 and 10 mg/kg to rats.

As previous research reported, almost no CFZ was detected in plasma 30 min after administration with an initial half-life (T1/2,initial) of <20 min.38 By utilizing the same solvent system composed of SBE--CD to solubilize CFZ (Sb-CFZ), a similar T1/2,initial was observed in this study, thereby confirming the suitability of the pharmacokinetic study conducted in this research. As such, a slower terminal elimination rate (T1/2,initial) observed for administration of CFZ/HSA NPs compared to that for administration of Sb-CFZ indicates that encapsulation of CFZ with HSA somewhat protected CFZ from elimination in plasma leading to a longer T1/2,initial. With a longer T1/2,initial, it was expected to have a higher AUC0-1h as Table 3 demonstrates.

Compared to Sb-CFZ, the combined administration of the two solvent-based formulations (Sb-CFZ+Sb-PTX) resulted in a similar AUC0-1h for CFZ but with a slightly lower T1/2,initial for the CFZ distribution into tissue compartments. This might indicate that drugdrug interactions exist between CFZ and PTX that are dissolved in solvent as free solubilized forms leading to an influence on the elimination rate of CFZ but not on the AUC. On the other hand, combined administration of the two HSA NP formulations (CFZ/HSA NP+PTX/HSA NPs) could have resulted in significant influences on both T1/2,initial and AUC0-1h for CFZ compared to those for CFZ/HSA NPs. Fortunately, although administration of CFZ/PTX co-loaded HSA NPs (CFZ/PTX/HSA NPs) led to a lower AUC0-1h than that for CFZ/HSA NPs, a higher AUC0-1h than those for Sb-CFZ and Sb-CFZ+Sb-PTX was observed. This also implies that co-encapsulation of CFZ and PTX in HSA with the simultaneous protection of CFZ and PTX by HSA might minimize drugdrug interactions that existed in the plasma compartment between CFZ and PTX when presented in free forms.

Since a greater difference existed between AUC0-2h and AUC0-24h for all the various formulations of PTX administered as shown in Table 4, pharmacokinetic parameters of T1/2,terminal (h) and AUC0-24h (hg/mL) were selected for comparison. Results in Table 4 indicate that T1/2,terminal (h) and AUC0-24h (hg/mL) for PTX after administration of Sb-PTX were shorter and much higher, respectively, than those for administration of PTX/HSA NPs (7.311.84 vs 12.890.65 min and 6.1712.018 vs 2.5110.497 hg/mL), while they both insignificantly differed from those for administration of Sb-CFZ+Sb-PTX (7.311.84 vs 9.081.92 min and 6.1712.018 vs 7.00402.082 hg/mL). On the other hand, T1/2,terminal (h) and AUC0-24h (hg/mL) for PTX after administration of PTX/HSA NPs greatly differed from those for administration of CFZ/HSA NP+PTX/HSA NPs (12.890.65 vs 13.371.41 min and 2.5110.497 vs 1.9580.824 hg/mL), while they were longer and slightly higher, respectively, than those for administration of CFZ/PTX/HSA NPs (12.890.56 vs 6.540.60 min and 2.5110.497 vs 1.7130.520 hg/mL).

It was reported that the administration of ABI-007 (Nab-paclitaxel or Abraxane) to Sprague-Dawley rats was associated with significantly higher CL and V of PTX compared to Taxol (Sb-PTX) resulting in a shorter T1/2,terminal (h) with a reduction in the AUC0-24h.35 This was attributed to the fact that the initial dilution volume and the central V were higher for PTX formulated as ABI-007 than for PTX formulated as Taxol resulting from Cremophor (as the solubilizing agent used in solvent-based formulations) preventing the distribution of PTX to the circulation and into tissues. What we observed in the comparative pharmacokinetic analysis performed in this study conformed to data in the literature, which showed that T1/2,terminal and AUC0-24h for PTX after administration of Sb-PTX and Sb-CFZ+Sb-PTX were both shorter and much higher, respectively, than those for administration of the albumin-bound counterpart of PTX/HSA NPs and CFZ/HSA NPs+PTX/HSA NPs, while T1/2,terminal and AUC0-24h for PTX after administration of Sb-PTX and PTX/HSA NPs both insignificantly differed from those with combination administration of either solvent-based or albumin-bound counterparts of Sb-CFZ+Sb-PTX and CFZ/HSA NPs+PTX/HSA NPs. Nevertheless, the administration of the co-loaded HSA NP formulation of CFZ/PTX/HSA NPs seemed to result in an even shorter T1/2,terminal (6.540.60 vs 12.890.65, 13.371.41 min) but not increasing AUC0-24h (1.7130.520 vs 2.5110.497, 1.9580.824 hg/mL) for PTX compared to that for administration of PTX/HSA NPs and CFZ/HSA NP+PTX/HSA NPs. The underlying reason for this discrepancy is currently unclear.

The anti-tumor efficacies of drug-loaded HSA NPs were evaluated on MIA Paca-2 cell-xenograft mice. At 14 days after inoculation when tumor volumes had reached 150 mm3, mice were intravenously administered saline, Sb-CFZ, CFZ/HSA NPs, Sb-PTX, PTX/HSA NPs, Sb-CFZ/Sb-PTX (1:2), CFZ/HSA NPs+PTX/HSA NPs (1:2), or CFZ/PTX/HSA NPs. The administration of each formulation was performed on days 0, 1, 7, 8, 14, and 15. Tumor volumes and BWs were assessed three times a week. Tumor growth profiles after administration of the various formulations plotted against time are shown in Figure 5A. TGI (%) compared to the control saline groups was calculated on day 21 after drug administration and on day 46 at termination of the study, and the results are illustrated in Figure 5B. All formulations expressed a greater suppression of tumor growth on both days 21 and 46 than that of saline (2301 mm3). Values of TGI (%) on days 21 and 46 for the Sb-CFZ group showed no improvement compared to the CFZ/HSA NP group, whereas those for the Sb-PTX group showed greater suppression than those for the PTX/HSA NP group on both days 21 and 46. Further, values of TGI (%) on days 21 and 46 for the two combined groups (Sb-CFZ+Sb-PTX and CFZ/HSA NPs+PTX/HSA NPs) all showed increases in TGI (%) compared those of each respective individual group (Sb-CFZ+Sb-PTX vs Sb-CFZ and Sb-PTX and CFZ/HSA NP+PTX/HSA NPs vs CFZ/HSA NPs and PTX/HSA NPs). The results confirmed that a synergic effect on the treatment of MIA PaCa-2 tumors was observed for the combination of CFZ and PTX at a 1:2 ratio regardless of whether Sb-CFZ+Sb-PTZ or CFZ/HSA NP+PTX/HSA NPs were examined. Although only a slight increase in TGI (%) was observed on day 21 but not on day 46 for CFZ/PTX/HSA NPs compared to those for Sb-CFZ+Sb-PTX and CFZ/HSA NPs+PTX/HSA NPs (CFZ/PTX/HSA NPs: 110.20%8.39% Sb-CFZ+Sb-PTX: 89.80%21.19%, CFZ/HSA NP+PTX/HSA NPs: 78.38%16.09%), over 100% of TGI means that tumors had obviously shrunk after treatment with CFZ/PTX/HSA NPs. This seems to indicate that the combination of CFZ and PTX at a 1:2 ratio encapsulated in HSA NPs synergistically improves tumor growth inhibition of MIA PaCa-2 cells.

Figure 5C further shows the weights (g) of tumors excised after tumor-bearing mice were sacrificed on day 46. It clearly shows that, compared to saline (2.330.52 g), there was a significant anti-tumor efficacy with any combined formulations of CFZ and PTX at a 1:2 ratio of Sb-CFZ+Sb-PTX (0.690.12 g), CFZ/HSA NP+PTX/HSA NPs (0.910.33 g), and CFZ/PTX/HSA NPs (0.750.11 g, all p<0.05), but there were no statistically significant differences in tumor weights among the three combined formulations. This further verifies that synergistic improvement in tumor inhibition is achievable with a combination of CFZ and PTX at a 1:2 ratio loaded into solvent-based or HSA NPs. Figure 5D also reveals that the decreases in BWs of mice after administration of various formulations were all smaller than 20% for the 46-day observation period. However, a greater decrease in BW of mice was observed at several time points with the administration of Sb-CFZ+Sb-PTX. This implies that the greater decrease in BW of mice might be attributed to a higher toxicity of solvents used in the solvent-based formulations compared to HSA used in the HSA NP formulations. It could be concluded that the combination therapy of CFZ and PTX at a 1:2 ratio co-loaded in HSA NPs (CFZ/PTX/HSA NP) demonstrated optimal synergistic improvement in the growth inhibition of MIA PaCa-2 cells with less systematic toxicity.

To examine the biodistribution of CFZ and PTX in tumors and major organs, C.B-17 SCID mice bearing MIA CaPa-2 tumors were injected with a single IV dose of various formulations including three solvent-based drugs (Sb-CFZ, Sb-PTX, and Sb-CFZ+Sb-PTX), and four drug-loaded HSA NPs (CFZ/HSA NPs, PTX/HSA NPs, CFZ/HSA NPs+PTX/HSA NPs, and CFZ/PTX/HSA NPs) with respective dosing amounts of CFZ and PTX equivalent to 5 and 10 mg/kg. Tumor tissues and major organs were harvested at 2 or 8 h post-injection, processed to make tissue homogenates, and subsequently analyzed with respect to CFZ and PTX levels by LC-MS/MS, and results are demonstrated in Figure 6. For the biodistribution of CFZ in tumor tissues as shown by Figure 6A (2 h) and 6B (8 h), both Sb-CFZ and CFZ/HSA NP groups presented insignificant difference in CFZ levels, but both displayed significantly higher levels of CFZ than those for the Sb-CFZ+Sb-PTX group at 2 h post-dosing with an accompanying decline in the CFZ level at 8 h post-dosing for those formulations examined. An undetectable CFZ level was seen in tumors for both the CFZ/HSA NP+PTX/HSA NP and CFZ/PTX/HSA NP groups at 2 h post-dosing and for those of the Sb-CFZ+Sb-PTX, CFZ/HSA NP+PTX/HSA NP, and CFZ/PTX/HSA NP groups at 8 h post-dosing. However, the CFZ level in tumors did not seem to be correlated with the tumor growth inhibition rate (TGI %) as revealed by Figure 5B. Similar patterns of CFZ biodistributions in these major organs examined for all formulations as distributed to tumors were observed with predominant distribution to the spleen at 2 h post-dosing with an accompanying decline in the CFZ level at 8 h post-dosing for the formulations examined. Similarly, the least or undetectable CFZ levels were shown in those major organs examined for both the CFZ/HSA NP+PTX/HSA NP and CFZ/PTX/HSA NP groups at 2 and 8 h post-dosing. Since quite lower levels of CFZ (<80 ng/g) were observed in those major organs, systemic toxicity caused by the presence of CFZ might not be highly anticipated.

Figure 6 Tissue distributions of carfilzomib (CFZ) and paclitaxel (PTX) at 2 (A and C) and 8 h (B and D), respectively, after intravenous administration of solvent-based (Sb)-CFZ, Sb-PTX, CFZ/human serum albumin (HSA) nanoparticles (NPs), PTX/HSA NPs, Sb-CFZ+Sb-PTX (1:2), CFZ/HSA NPs+PTX/HSA NPs (1:2), or CFZ/PTX/HSA NPs (equivalent to 5 mg/kg CFZ and 10 mg/kg PTX in each mouse). *p<0.05.

For the biodistribution of PTX in tumors as shown by Figure 6C (2 h) and D (8 h), the Sb-PTX group presented a statistically significantly higher level of PTX than those for the PTX/HSA NP and CFZ/PTX/HSA NP groups at 2 h post-dosing (40.005.91 vs 19.744.51 and 5.722.85 g/g), while those for PTX/HSA NPs and CFZ/HSA NP+PTX/HSA NPs were similar, but both were statistically higher than that for CFZ/PTX/HSA NPs (19.744.51 21.609.54 vs 5.722.85 g/g). However, a slight increase was shown in the PTX level biodistributed in tumors for the Sb-PTX group at 8 h post-dosing compared to that at 2 h post-dosing, whereas those for the remaining formulations at 8 h post-dosing (PTX/HSA NPs, Sb-CFZ+Sb-PTX, CFZ/HSA NPs+PTX/HSA NPs, and CFZ/PTX/HSA NPs) were still statistically significantly lower than that for the Sb-PTX group but showed insignificant differences among them (50.528.60 vs 20.6215.08, 15.066.84, 15.753.88, and 12.983.93 g/g). Similar to CFZ as described above, the PTX level distributed to tumors did not seem to be correlated with the tumor growth inhibition rate (TGI %) as revealed by Figure 5B. Similar patterns of PTX biodistributions in the major organs examined for all formulations as those distributed to tumors were observed with predominant distribution to the liver, spleen, and kidneys at 2 h post-dosing with accompanying significant declines in PTX levels at 8 h post-dosing for those formulations examined. Since quite higher levels of PTX were detected in the liver (134.721.9 g/g at 2 h and 28.9212.62 g/g at 8 h) after administration of Sb-PTX compared to those for the other formulations examined, the higher grade of systemic toxicity caused by administration of Sb-PTX in the presence of this amount of PTX in the liver might be highly expected. It is worth noting that although both 2 and 8 h accumulations of PTX in tumors with the administration of Sb-PTX were 2~3 times more than those for the two combined groups (Sb-CFZ+Sb-PTX and CFZ/HSA NPs+PTX/HSA NPs), TGI (%) values on days 21 and 46 for the two combined groups (Sb-CFZ+Sb-PTX and CFZ/HSA NPs+PTX/HSA NPs) as revealed above all showed increases in TGI (%) compared to that for the Sb-PTX group. This seems to further confirm that the synergistic improvement in tumor growth inhibition is achievable with a combination of CFZ and PTX at a 1:2 ratio loaded into solvent-based or HSA NPs with minimal systemic toxicity.

It was concluded that the effective combination therapy of pancreatic cancer was enabled with treatment of CFZ and PTX co-loaded HSA NPs, which was prepared by a simple one-pot reverse self-assembly method developed in this study. The one-pot reverse self-assembly method was novel and able to optimally prepare HSA NPs loaded with hydrophobic drugs by adjusting the drug/HSA ratio and homogenization process parameters. Without using any hazardous or toxic solvent during preparation of drug-loaded HSA NPs, the one-pot reverse self-assembly method could claim to be environmentally friendly with the ability to co-encapsulate two chemodrugs in HSA NPs with the optimal ratio for synergistic therapy to inhibit tumor growth and minimize systemic toxicity compared to monotherapy. With the related data in this study, it might be able to construct a platform for combination therapy in the future.

This animal experiment was approved by the Institutional Animal Care and Use Committee of Taipei Medical University (Approval No.: LAC-2018-0419) in compliance with the Taiwanese Animal Welfare Act.

This work was supported by the Ministry of Science and Technology, Taiwan, ROC, under grant no. 107-2314-B-038-035-MY3, 108-2314-B-264-001- and 110-2221-E-264-002-.

The authors report no conflicts of interest with respect to this work.

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Combination Therapy of Carfilzomib and Paclitaxel for PACs | IJN - Dove Medical Press

This article was originally published here

Int J Nanomedicine. 2021 Sep 24;16:6575-6591. doi: 10.2147/IJN.S273062. eCollection 2021.

ABSTRACT

Public awareness of infectious diseases has increased in recent months, not only due to the current COVID-19 outbreak but also because of antimicrobial resistance (AMR) being declared a top-10 global health threat by the World Health Organization (WHO) in 2019. These global issues have spiked the realization that new and more efficient methods and approaches are urgently required to efficiently combat and overcome the failures in the diagnosis and therapy of infectious disease. This holds true not only for current diseases, but we should also have enough readiness to fight the unforeseen diseases so as to avoid future pandemics. A paradigm shift is needed, not only in infection treatment, but also diagnostic practices, to overcome the potential failures associated with early diagnosis stages, leading to unnecessary and inefficient treatments, while simultaneously promoting AMR. With the development of nanotechnology, nanomaterials fabricated as multifunctional nano-platforms for antibacterial therapeutics, diagnostics, or both (known as theranostics) have attracted increasing attention. In the research field of nanomedicine, mesoporous silica nanoparticles (MSN) with a tailored structure, large surface area, high loading capacity, abundant chemical versatility, and acceptable biocompatibility, have shown great potential to integrate the desired functions for diagnosis of bacterial infections. The focus of this review is to present the advances in mesoporous materials in the form of nanoparticles (NPs) or composites that can easily and flexibly accommodate dual or multifunctional capabilities of separation, identification and tracking performed during the diagnosis of infectious diseases together with the inspiring NP designs in diagnosis of bacterial infections.

PMID:34602819 | PMC:PMC8478671 | DOI:10.2147/IJN.S273062

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Recent Advances in the Use of Mesoporous Silica Nanoparticles for the Diagnosis of Bacterial Infections - DocWire News

The word nano refers to the length scale (one nanometre is one-billionth of a metre) that is one thousand times smaller than the micro scale, the scale that was traditionally associated with the electronics industry. Viruses and DNA are examples of natural objects on the nano scale; in contrast a human cell can appear enormous.

The term nanotechnology refers to the engineering, measurement and understanding of nano-scaled materials and devices. Manipulating matter atom by atom and creating features on the atomic or nano scale is now a proven technology and there is an ever growing catalogue that utilises nanotechnology.

Nanotechnology represents an entire scientific and engineering field, broadly within Materials Science and Engineering, and not just a single product or even group of products. As a consequence of this there are several different types of nanotechnology, and many applications associated with each type. There are also several other types of nano-sized objects which exist in our environment, both natural and unnatural such as films and coatings, embedded nanotechnology, biologically natural, biological nanotechnology, natural particles, manufactured particles, nano-electrical mechanical systems.

Building on current nanotechnology-enabled applications in areas as diverse as consumer electronics, medicine, energy, water purification, aerospace, automotive, infrastructure, sporting goods, textiles, and agriculture, the nanotechnology research underway today will enable entirely new capabilities and products. Nanotechnology also underpins key industries of the future. For example, new architecture and paradigms exploiting nanotechnology are providing the foundation for artificial intelligence (AI), quantum information science (QIS), next-generation wireless communications, and advanced manufacturing.

While advances in modern electronics have long been at the nanoscale, new nanomaterials and designs will ensure the continued strength of the semiconductor industry, which powers computing, e-commerce, and national security. Nanotechnology also enables the rapid genomic sequencing and sensing required to advance medicine and biotechnology. Nanotechnology R&D has enabled early detection of emerging diseases and will lead to the treatments of the future. Past investments in nanotechnology research and development have provided a foundation to support the response to the Covid-19 pandemic. Nanotechnology-enabled applications include vaccines, sensors, masks, filters, and antimicrobial coatings.

Examples of nanotechnology innovations are: a highly sensitive wearable gas sensor; nanoparticles absorbed by plants to deliver nutrients; durable, conductive yarns made with MXene; electrodes that incorporate nanoparticles and enable the conversion of sunlight to hydrogen fuel; nano-engineered pores in a membrane for water filtration; drug-loaded nano particles carried by red blood cells; and the first programmable memristor computer, enabling low-power AI applications. Nanotechnology advances are impacting a variety of other sectors including consumer electronics, aerospace, automotive, infrastructure, sporting goods, and agriculture.

Research Infrastructure

The research infrastructure, including physical and cyber resources as well as education and workforce development efforts, is critical to support the entire funding ecosystem (National Nanotechnology Initiative), and agencies will continue to invest in these important areas. Agencies use a wide variety of mechanisms to support the research infrastructure, including Centre grants, instrumentation development or acquisition programmes, training grants, fellowships, and collaborative programmes that support workforce development.

Career opportunities

The scope and application of nanotechnology is tremendous. Indian engineering and science graduates are increasingly opting for nanotechnology. Right from medicine, pharmaceuticals, information technology, electronic, opto-electronics, energy, chemicals, advanced materials to textiles, nanotechnology has its applications. Nanotechnology provides job opportunities in health industry; pharmaceutical industry; agriculture industry; environment industry; food and beverage industry as well in government and private research institutes.

Skills

One needs to have a diehard passion for research, especially to find out new structures in the field of nanotechnology. It is important to have sound analytical skills, along with a scientific bent of mind. Analysing and interpreting skills are a necessity in this field and also to accept failures in experiments as a challenge. Other necessary skills which are required are: Good mathematical and computer programming skills; adequate laboratory training for expert handling of advanced equipment; ability to learn and adopt new techniques; have a systematic way of working; a natural propensity for research work; keep track of the latest scientific news, books and research magazines; a good background of physics, chemistry, medicine, electronics and biotechnology

Job Prospects

A lot of job opportunities and a research career exists in the areas of nano-device, nano-packaging, nano-wires, nano-tools, nano-biotechnology, nano-crystalline materials, nano-photonics and nano-porous materials to name a few. It is estimated that around three million nanotechnology skilled workforce will be required worldwide by 2021. Many government institutes and Indian industries have focused on nano-materials. It is also estimated nano-technology will create another five million jobs worldwide in support fields and industries. A professional in the field of nanotechnology can easily find lucrative jobs in most of fields.

Since nanotechnology is a special branch that essentially combines physics, chemistry, biology, engineering and technology, it is opening up job prospects for students specialising in these subjects. The career opportunities in the fields of nanoscale science and technology are expanding rapidly, as these fields have increasing impact on many aspects of our daily lives.

A professional in the field of nanotechnology can easily find viable career opportunities in various sectors. They can work in the field of nano-medicine, bio-informatics, stem cell development, pharmaceutical companies, and nano toxicology and nano power generating sectors.

The major areas for the development of applications involving nanotechnology are medical and pharmaceuticals, information technology, electronics, magnetics and opto-electronics, energy chemicals, advanced materials and textiles.

Nanotechnology has varied applications in drug delivery to treat cancer tumours (without using radiotherapy and chemotherapy), solar energy, batteries, display technologies, opto-electronic devices, semiconductor devices, biosensors, luminous paints, and many others. A major challenge in this emerging field is the training for a new generation of skilled professionals.

An abundance of job opportunities awaits candidates with an MTech in Nanotechnology from India and abroad. Indian industry has focused on nanomaterials and many scientific institutions have started research and development activities in the field. The CSIR has set up 38 laboratories, across the country, to carry out research and development work in this field. Those with a PhD in Nanotechnology will have vibrant opportunities in the R&D sectors.

It is a perfect career for those who have a scientific bent of mind and a passion for studying and experimenting with the minutest molecules. Students with a science and engineering background and even mathematics with a physics background can pursue Nanotechnology as a career. Candidates with MTech in Nanotechnology are in great demand both in India and abroad.

(The writer is Associate Professor, Department of Physics and Nanotechnology, SRM University)

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Nanotech opens up job options in variety of industries - BL on Campus

Introduction

The World Health Organization (WHO) affirmed COVID-19 as a pandemic on 11 March 2020 but earlier to this the Chinese government confirmed the first outbreak of Coronavirus disease 2019 (COVID-19) in Wuhan on 31December 2019. The state-wise lockdown, which was imposed in India due to the second wave of the novel coronavirus pandemic, affected people belonging to every economic stratum. In India, till now (9 July 2021), there have been 30,752,950 confirmed cases of COVID-19 with 405,939 deaths reported to the WHO. COVID-19 cases are rapidly rising globally of which the first case was registered on 21February 2020 in Italy. Meanwhile in India, case numbers have risen, and community transmission was officially declared by government in October 2020. Life is deeply affected by COVID-19 even for the ones who are not infected as isolation, contact restrictions and economic shutdown have changed the social and economic scenario of India. Vast populations and crowded settlements have increased the number of cases in China, Europe, USA and India. Countries with dense populations and robust travel history will increase the problem of decision-making authorities if testing is limited or disproportionate. The WHO has made projections of 3.5 beds per 1000 population1 but many countries have only 1.3 beds per 1000 population in hospitals which is again the concern of government. As the pandemic is growing in stages, this review assesses the prospects these stages might have on the Indian population as it highlights some key challenges for treatment and research related to antiviral drugs.

Cases were initially spread by migrants, overseas visitors, and some others who were in contact with these infected persons, and to control this spread lockdowns were called by various countries including India. The situation seemed to be under control due to the lockdown, but due to a religious gathering in New Delhi, which led to the human-to-human transmission of COVID-19, a sudden horrific increase in COVID-19 cases occurred. Initially, most individuals who came into contact with such infected individuals were unaware of the effects of the virus in their bodies. To sustain the countrys economy, unlocks were called by the Indian Government in multiple phases, therefore, the persons who were unaware that they were carrying the virus spread it many more healthy persons. However, preventive measures including social distancing, quarantine and isolation techniques had been taken globally and have proven effective in the absence of drug treatments and other approaches. Adults (ages 50 and over), and people with comorbidities can have higher chances of becoming severely ill with COVID-19 and contribute to the largest portion of all deaths worldwide among infected cases.2,3

In India, the overall numbers dying constantly increased, amongst them a lot of the demise circumstances pointed to a particular age-group of aged folks.4 In India, among the total COVID-19 cases (30,752,950) and total deaths (405,939) till 9 July 2021, 90% were older than 40 years. Overall, people in the age group of 40 years and greater, have suffered the major impact of the current COVID-19 eruption and are more vulnerable.5,6 The massive loss of people in the workforce is likely to have devastating social and economic consequences.

The basic measures adopted worldwide include maintenance of hand hygiene, avoiding close contact, using face masks, disinfection and monitoring health.7 The ongoing COVID-19 pandemic has once again brought the benefits of appropriate hand hygiene (hand washing and use of alcohol-based hand-sanitizers) to the centre stage. Since hand washing is not a feasible and available option at all times, the use of alcohol-based hand-sanitizers (hand rubs) has been recommended by health organizations, when hands are not visibly soiled. These sanitizers act as a powerful, fast acting and effective solution with broad antimicrobial range.7 Hands act as a medium for exchange of microbes between the organism and its environment. The skin of the hands harbours a variety of organisms ranging from commensal to potential pathogens. Therefore, adequate hand hygiene can greatly reduce disease transmission. The most commonly used agents for hand disinfection are hand-sanitizers. There are two major types of preparations available: alcohol-based and alcohol-free. The alcohol-based ones, known as alcohol-based hand rubs (ABHRs), typically have ethyl alcohol (ethanol), isopropanol, or n-propanol at concentrations between 60 to 95% alcohol.8 The alcohol-free preparations usually contain quaternary ammonium compounds (benzalkonium chloride or benzethonium chloride). However, these have been found to be less effective and have a risk of contributing to antimicrobial resistance (AMR), hence are not recommended by CDC.

The CDC has recommended the use of ABHRs and hand washing to fight the COVID-19 pandemic. This is due to the structural characteristics of coronaviruses, which are enveloped viruses with lipid bilayer and are easily inactivated by alcohol. A combination of factors such as inappropriate formulations, excessive/repeated usage of hand sanitizers during this pandemic will have far reaching consequences. These may range from emergence of situation like alcohol tolerance and antimicrobial resistance (AMR), disturbance of normal microflora, and product toxicity. Similar to antibiotics, excessive or repetitive application of alcohol through hand-sanitizers has the potential to act as a selection pressure for the emergence of new microbial species tolerant to high alcohol concentrations.9

Taking note of the repetitive use of ABHRs, Professor Tim Stinear from the Peter Doherty Institute for Infection and Immunity remarked

Anywhere we repeat a procedure over and over again, whether its in a hospital or at home or anywhere else, youre giving bacteria an opportunity to adapt, because thats what they do, they mutate. The ones that survive the new environment better then go on to thrive.

He further added that the risk increases when appropriate guidelines are not followed.10

Eliminating the normal microflora of the skin by repeated use of hand-sanitizers may eventually deprive the skin of the protection offered by these commensals. Long term use of personal protective equipment along with frequent hand hygiene was responsible for high rate of skin damage in 97% of respondents while frequent hand hygiene was attributed with increased risk of hand skin damage.11

The world has joined hands with parallel efforts for the production of vaccines in opposition to COVID-19 pandemic.

A densely populated area like Ladakh has set an example for implementation in the Guidelines for hygiene and sanitation during the era of COVID-19 pandemic by setting up Foot-Operated Washing Station, implemented at the Indian Astronomical Observatory (IAO), Hanle. Having one of the worlds highest located sites for optical, infrared and gamma-ray telescopes operated by the Indian Institute of Astrophysics (IIA), Bengaluru, IAO12 has one in all the worlds highest set sites for optical, infrared and gamma-ray telescopes.

Antiviral nano-coating and new nano-based material for use in Personal Protective Equipment (PPE) was invited by The Department of Science and Technology (DST) using the Science and Engineering Research Board (SERB) portal, scale up for which could be done by partnering industry or start-up. India could be supported greatly by such nano-coatings technology to fight against COVID-19 pandemic. N-95 respirator, PPEs kits and triple-layer medical masks could be prepared from antiviral nano-coatings for safeguarding healthcare workers.13

Patients that showed flu-like symptoms was screened and detected for COVID-19 through indigenous company Mylab Discovery Solutions through the development of PCR-based molecular diagnostic kit.

TDB will try to boost the production process of kits so that present capacity could increase from 30,000 tests per day to one lakh tests per day. This automation by company could be achieved within the next few months. Considering the national emergency COVID-19 kit will be deployed by ICMR and CDSCO.14

As the demand increased, production of sanitizers have seen a boom amid coronavirus outbreak. Owing to which alcohol-based herbal sanitizer was developed by NBRI under Council of Scientific and Industrial Research (CSIR)-Aroma Mission as per the World Health Organisation (WHO) guidelines. Apart from having 60% of isopropyl alcohol for killing germs it has essential oil from Tulsi as natural antimicrobial agent. It is not only last for 25 minutes but also prevents skin from dehydrating. Herbal sanitizer has been found to be effective against the pathogen (Staphylococcus epidermidis).15

The Council of Scientific and Industrial Research (CSIR) is leaving no stone unturned in the battle against novel coronavirus. Repurposing of existing drugs is one of the strategies deployed by CSIR. The Council is implementing this strategy by evaluating an existing drug (Sepsivac, that available commercially) that is used for treating gram-negative sepsis patients. Both Gram-negative sepsis patients and critically ill COVID-19 patients, exhibit the altered immune response and a massive change in the cytokine profiles. Cytokines are produced in response to an infection and they are essential for host defence against pathogens. There are six types of cytokines, which belong to different families and the mixtures of cytokines, called cytokine profiles. One of the significant contributors to death by COVID-19, has shown the heightened immune response, called a cytokine storm. The immune system starts attacking both infected as well as uninfected cells and unable to discriminate between a friend and a foe, leading to tissue damage which resulting in sepsis. This drug (Sepsivac) modulates the immune system of the body and thereby inhibits the cytokine storm leading to reduced mortality and faster recovery.16

ICMR releases advisory for use of Cartridge-based Nucleic Acid Amplification Test (CBNAAT) using Cepheid Xpert Xpress SARS-CoV-2, effective from 19 April 2020.17

Indias first antibody-based testing kit was developed by NuLife Consultants and Distributors Pvt. Ltd, New Delhi which takes only fifteen minutes to yield accurate results. It is launched in two weeks and regular production has also started it was approved by the Indian Council of Medical Research (ICMR).18 The new finger prick kit will provide adequate access to cost-effective testing.

Home screening test kit for COVID-19 was launched by Bione with easy-to-use kit displays after approval from the requisite medical regulatory authorities.

In a get through development, the Company has devised the screening kit which can provide respite from the impending fear of the contagion. It will foster timely detection of the disease while acting as a preventive tool for others in proximity to the user, by isolating the carrier immediately. The kit is priced between `20003000 depending upon the global supply, to increase its affordability for the masses. Under normal circumstances, the ready-to-use kits can be received within 23 days of placing the order at their platform. To initiate an effective screening tool for mass screening, the organisation is also in talks to provide bulk orders for early detection.19

Against COVID-19 drugs and experimental molecule are being prepared. SARS-CoV-2 is a single stranded RNA enveloped virus. The angiotensin-converting enzyme 2 (ACE2) receptor of the host cell binds to the spike (S) protein of the viral structure. The host type 2 transmembrane serine protease, TMPRSS2 facilitates the S protein.20 Once the virus enters the host, it starts synthesizing RNA through its RNA dependent RNA polymerase enzyme, which is then translated to products. Structural proteins facilitate the assembly and release of viral particles.21,22

During viral life cycle, chemotherapy is available of various potential targets. There are many non-structural protein promising drug targets which resembles with other coronaviruses (SARS-CoV and MERS-CoV) such as 3-chymotrypsin like protease, papain like protease and RNA-dependent RNA polymerase. Various molecules and their targets are represented in Figure 1.

Figure 1 Mechanism of various drugs/molecules on COVID-19 disease.

Chloroquine and hydroxychloroquine used in prevention and treatment of malaria and chronic inflammatory diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA).23 CQ and HCQ are reliable anti-malarial drugs approved by FDA, which shows positive response against SARS-CoV-2 infections and hence used for the treatment of COVID-19 patients by clinicians.2426 It inhibits the entry of the virus by either altering the configuration of structure of cell receptors or by compete to bind with cellular receptors.27 The glycosylation of ACE-2 cellular receptors can amend by CQ/HCQ which is needed for entry of SARS-CoV-2. Apart from that CQ/HCQ can also prevent the attachment of SARS-CoV-2 to the host cells by decrease the synthesis of sialic acid.

The binding affinity of these drugs is better as compared to the S protein of SARS-CoV-2. Therefore it prevents attachment and entry of virus because of competitive binding of sialic acid and gangliosides present on surface pf target cell.28

In addition to the antiviral activity of CQ/HCQ, they have anti-inflammatory activity that may contribute to its efficacy in treating COVID-19 patients. Through the attenuation of cytokine production, these drugs also have immunomodulatory effects and inhibition of lysosomal and autophagy activity in host cells.24,29 In vitro activity of HCQ with lower EC50 for SARS-CoV-2 as compared to CQ after the growth of 24 hours (HCQ: EC50=6.14 M and CQ: EC50=23.90 M).30

A study from China reported which results in improved radiologic findings, enhanced viral clearance, and reduced disease progression by treating successfully with CQ on 100 COVID-19 cases.31 When treatment given to 6 patients, then it is observed that as compared to HCQ monotherapy (8/14, 57%) the combination of azithromycin with HCQ (6/6, 100%) results in numerically superior viral clearance.32

Other than these positive results, this study has many limitations like intolerance of medication, different viral loads between HQC combination and monotherapy and no safety outcomes are reported.

Another study of 30 patients in China shows there was no difference in virologic outcomes to HCQ plus standard of care (supportive care, interferon, and other antivirals). At 7th day virologic clearance was similar with clearance for the HCQ plus standard of care group and standard care group ie 86.7% vs 93.3% respectively, (P>.05).33 Currently, for COVID-19 treatment several RCTs of both CQ and HCQ examining their roles. To treat COVID 19 500 mg dose of CQ orally once or twice daily is advised.8,9

However, there is shortage of data regarding the optical dose to ensure efficiency of CQ For HCQ, daily dose of 400 mg taken orally is recommended.34

Both the agents are well tolerated by patients with SLE and malaria as demonstrated by their experiences and they can cause rare and serious adverse effect (>10%) such as hypoglycemia, neuropsychiatric effects, and retinopathy.

Lopinavir/ritonavir is FDA approved for treating HIV and it shows in vitro activity against coronavirus by inhibiting 3-chymotrypsin like protease.35 The therapy during early peak viral replication phase (initial 710 days) is important because delayed medication with lopinavir/ritonavir had no effective outcomes.36,37

Although many RCTs of lopinavir/ritonavir examine their role, limited role for lopinavir/ritonavir in COVID-19 treatment is suggested through current data.38

Recent RCT shows that approximately 50% of patients experienced an adverse effect under the lopinavir/ritonavir therapy and 14% of patients stop therapy due to adverse effects on gastrointestinal region. In several COVID-19 investigational trials, alanine transaminase elevations are exclusion criterion. Hepatotoxicity induced by lopinavir/ritonavir could limit patients ability to access these drugs.39

The activity of darunavir is demonstrated in vitro cell models against SARS-CoV-2. With these drugs there is no clinical data is available in COVID 19, but in China RCT of darunavir/cobicistat is going on.40,41 Ribavirin is a analogue to guanine which inhibits viral RNA-dependent RNA polymerase and used as best candidate for treatment of COVID 19.

However, it has limited in vitro activity against SARS-CoV and high doses is required to prevent viral replication (e.g., 1.2 g to 2.4 g orally every 8 hours) and combination therapy. For nCoV treatment no evidence exists for inhaled ribavirin..42 Generally ribavirin is used in combination with interferons in the treatment of MERS, no visible effect is shown on clinical outcomes. A lack of clinical data with ribavirin for treatment of COVID 19, means its therapeutic role must be extrapolated from other nCoV data.43,44 The high doses used during trials SARS resulted in hematologic toxicity and hemolytic anemia in more than 60% of patients. Similar safety concerns were seen in MERS trial, with 40% of patients taking ribavirin with interferon requiring blood transfusions. 75% of patients experienced transaminase elevations while taking ribavirin for SARS. Ribavirin is a teratogen and prescribed as not to be used pregnancy.45,46

It is a nucleoside reverse - transcriptase inhibitor that is worthy in clinical trial against COVID-19. It acts as an inhibitor of RNA-dependent RNA polymerase (RdRp)47 and in SARS-CoV and MERS-CoV infections its pharmacokinetics and characteristics have been studied.48 It inhibits the viral genomic replication and production by disturbed reading due to alteration in the function of viral exonuclease.49

Therefore it can suggested for COVID 19 patients to prevent severity of disease progression such patients are taken to phase 3 trials to check the therapeutic efficiency of remdesivir.50

Favipiravir (T705) is considered as RdRp inhibitor as it is an analog to guanine nucleotide (a derivative of pyrazine carboxamide).51 Initially it was used against influenza but because of its large spectrum antiviral properties, it attracted more attention for treatment of COVID 19.52

An in silico study showed that as compared to lopinavir, atazanavir bound more strongly to the active site of SARS-CoV-2 MPro and an in vitro study found that replication of SARS-CoV-2 inhibited by atazanavir.53

Oseltamivir is used for treatment of influenza because it acts as a neuraminidase inhibitor. It has no data against SARS-CoV-2. Initially in China during the COVID-19 outbreak until the discovery of SARS-CoV-2 as the cause of COVID-19 a large proportion of patients were treated with oseltamivir therapy because outbreak occurred in influenza season.

Once influenza has been excluded this agent has no role in the management of COVID-19.54 Umifenovir has a unique mechanism of action targeting the S protein interaction and inhibiting membrane fusion of the viral envelope. This agent is approved for treatment of influenza in Russia and China and treatment of COVID 19 patients started on the basis of in vitro data which shows its activity against SARS.

A study shows that 67 patients treated with Umifenovir for 9 days had a lower mortality rate and higher discharge rate compared with the patients who were not treated with this medication. This data cannot proof the efficiency of umifenovir, but for evaluating this agent further RCTs are going on in China.55.

For SARS-CoV-2 interferon- and - have been studied, due to their demonstrating activity against MERS by interferon-. Some interferons are listed as an alternative for combination therapy by Chinese guidelines. Traditionally other agents are used to demonstrate in vitro activity to inhibit SARS-CoV-2, but not limited to baricitinib, dasatinib, and cyclosporine. However it should be seen whether it provide protection for COVID 19 patients or not.56

Nitazoxanide has in vitro antiviral activity against MERS and SARS-CoV-2. It is used traditionally as an antihelminthic agent. More studies are required to check the antiviral activity and immunomodulatory effects of this agent. For treatment option for SARS-CoV-2 nitazoxanide is recommended.57 In Japan camostat mesylate is used for treatment of pancreatitis, it prevents cell entry through the host serine protease, TMPRSS2. For future research this mechanism provides an additional drug target.58

The ACE2 receptor is used by SARS-CoV-2 for entry into the host cell. This discovery has increased questions about whether ACE inhibitors and/or angiotensin receptor blockers may efficiently treat COVID-19 or either worsen disease. There are some conflicts if these provide protective effect to COVID-19 patients. Further research is pending for recommending therapy for patients already taking one of these agents.59,60

One of the main challenges in this pandemic is to develop multiple technology platforms for evaluation of agents/molecules against SARS-CoV-2 as this virus shows similarity with various other (Figure 1) corona viruses and shares similar binding receptors (ACE2) in humans (host).61 SARS-CoV-2 has ss-RNA genome of approximately 30 Kbp size and exhibits approximately 89% nucleotide similarly to SARS-CoV found in Chinese bats.20

For SARS-CoV 2 various technologies are being developed such as nucleic acid, replicating viral vector and non-replicating viral vector. New methods based on nucleic acid can facilitate rapid production because they do not need to be fermented. Experiments are conducted to ensure vaccination of larger population without any reduction in efficacy but also with improved immune response along with low dosages.62,63

As of January 2021, more than 200 vaccine candidates for COVID-19 are being tested. Among these almost 52 vaccines are approved for human trials and many other vaccines are in phases I/II and will soon enter phase III trials. Certain national regulatory authorities have nine authorized COVID-19 vaccines.

It represents a classic strategy for viral vaccinations. Finally, a codon deoptimization technology to attenuate the viruses is employed by Codagenix64 and is testing to develop vaccine against SARS-CoV-2, CodaVax-COVID. The inherent immunogenicity and ability to stimulate toll-like receptors (TLRs) is a major advantage of whole virus vaccines. This is especially an issue for coronavirus vaccines, given the findings of increased infectivity following immunization with live or killed whole virus SARS coronavirus vaccines.65

Subunit vaccines depend on producing immune response against S protein to inhibit its binding with host ACE2 receptor.65 Immunogenic virus-like nanoparticles produced by Novavax are based on recombinant expression of the S-protein66 while subunit vaccine consisted of a trimerized SARS-CoV-2 S-protein is developed by Clover Biopharmaceuticals by using their patented Trimer-Tag technology.67

For development of COVID-19 vaccines several major biotech industries have advanced nucleic acid vaccine platforms. Some modifications and formulation have improved nucleic acid performance in humans. This approach may lead to the first licensed nucleic acid based vaccine for humans.

Developing vaccine against the SARS-CoV-2 can cause distinct challenges. Various proteins of SARS-CoV-2 are used for developing proteins like S protein, N protein, M protein is the initial challenge. Developing a vaccine is a long process, starting from product development to the completion of phase III and clinical trials before marketing which takes several years.

Vaccine against COVID-19, known as CoroFlu is under process and its development and testing is done by Bharat Biotech in collaboration with international virologists and vaccine makers. One-drop COVID-19 nasal vaccine named CoroFlu, it is well tolerated in human trials during phase I and phase II. On the backbone of FluGens flu vaccine, CoroFlu has built a candidate known as M2SR. M2SR induces an immune response against the flu; it is a self-limiting version of the influenza virus. To induce immunity against the coronavirus in new virus, Kawaokas lab is trying to insert the gene sequences from SARS-CoV-2 into M2SR.68

To develop a vaccine for SARS-CoV-2, Zydus Cadila, an innovation-driven global pharmaceutical company, initiated a research program along with multiple teams. By reverse genetics the recombinant measles virus (rMV) is produced. It would express codon optimised proteins of the SARS-CoV-2 and provide long-term neutralising antibodies for protection from infection. The plasmid DNA vaccine, also has wide ranging capabilities in developing and manufacturing different vaccines for unmet needs. This is under supervision of the groups Vaccine Technology Centre in India.69

To develop a lead vaccine candidate for SARS-CoV-2 the Vaccine manufacturer Indian Immunologicals Ltd (IIL) has a research collaboration agreement with Australias Griffith University. As part of the cross-continental collaboration, using the latest codon de-optimisation technology Live Attenuated SARS-CoV-2 vaccine could be developed by scientists from IIL and the Griffith University. with a single dose administration this vaccine is expected to provide long protection with an anticipated safety profile for active immunization.70

Now the SII (Serum Institute of india) is preparing its mass production against the coronavirus, mixing out doses of the Covishield candidate vaccine which is being developed by the University of Oxford and the international biopharma company AstraZeneca. In India stage III clinical trials of Covishield are continuing. In the US, Brazil and South Africa the candidate vaccine is also being tested in various stages. Two million doses of the vaccine candidate has already produced over for use in testing by the SII. Recently SII announced a deal with Codagenix, US-based Biotech Company to help develop a vaccine candidate and it is expected that its trials starts by the end of 2020. Nasal COVID-19 vaccine candidate developed by Codagenix Inc. Dubbed the DX-005, manufacturing by SII has started.

After completing preclinical animal studies the coronavirus vaccine entered phase I clinical trials in the United Kingdom by the end of 2020. Bharat Biotech, a private firm collaborated with Indian Council of Medical Research (ICMR) is developing Covaxin. Covaxin has shown good efficacy is said by task force scientist Dr. Rajni Kant ICMR-COVID-19. Bharat Biotech is approved by The Drugs Controller General of India (DCGI) to perform Phase III clinical trials of Covaxin with certain conditions.

Russias president Vladimir Putin endorsed approval of SPUTNIK V (COVID-19 vaccine) that has not passed rigorous medical tests and could have numerous consequences. The effectiveness of the vaccine in response to providing active acquired immunity against COVID-19 and its possible adverse effects remain unknown. Therefore, the fear of vaccination in this particular case may be justified. However, endorsement of a potentially harmful vaccine will inevitably fuel public fears of other existing and future, properly developed, controlled and safe vaccines. Current level of public acceptability of immunization is already worrying, putting at serious risk the effectiveness of any future anti-SARS-CoV-2 vaccination programs, as it has been pointed out by Cornwall 2 and the French COCONEL Group 3. Independently from each other these groups provide evidence that it is a transatlantic phenomenon. Regardless of the suggested correlations between vaccination hesitancy and specific socioeconomic factors, it is clear that anti-vaccination movements are increasingly influential.71 Moreover, the problem is internationally valid, and the rise in the number of adults openly hesitant about routine childhood vaccination in many Western countries justifies the concern about public participation once the COVID-19 vaccine is available.72

In terms of collective immunity, vaccination effectiveness is based on its mass implementation; this may seriously undermine the efforts to protect societies against COVID-19 in the near future. High levels of COVID-19 vaccine hesitancy are reported even from countries severely affected by the pandemic. Only 49% of American respondents plan to vaccinate when the vaccine becomes available.73

Polish research confirms the strong COVID-19 vaccination hesitancy and its international character which is not directly related to the level of confidence in vaccination safety in general. Results of this Polish study show that 28% of adults would not vaccinate against SARS-CoV-2 if the vaccine became available. Alarmingly, a majority (51%) of the reluctant respondents indicated that their minds would not be changed if given information regarding vaccine safety or efficacy, or if threatened with heavy fines. Significantly fewer respondents (37%) supported COVID-19 vaccinations specifically than supported childhood vaccinations in Poland in general (78% in 2018).74 The vaccine hesitancy for the anticipated COVID-19 vaccine varied from very low (26% China) to very high (43%, Czechia, and 44%, Turkey). Surprisingly, the level of unwillingness to vaccinate against COVID-19 is in most countries much higher than regular vaccination reluctance, which varies between 3% (Egypt) and 55% (Russia). Such high levels of vaccination hesitancy may be detrimental to public health. According to current estimates, the benefits of herd immunity are achievable if 67% of the population is vaccinated.75,76

The most effective vaccination programs in the past effectively eradicated certain deadly diseases, such as smallpox which was achieved by combining the mandatory preventive vaccination programs with coordinated education efforts.77 Coronaviruses mortality rate is the highest among elders and people with comorbidities or conditions that affect their immune system. Some occupations have been identified as being the riskiest in terms of contracting COVID-19 such as health-care workers (dental hygienists, family practitioners, and nurses), transportation personnel (flying attendants, and school bus drivers), kindergarten, school teachers, fire fighters and restaurant personnel.78 Highest risk of death and highest risk of contraction should constitute the main criteria for mandatory vaccination. Mandatory vaccination will definitely trigger massive opposition especially bearing in mind the massive protests against social distancing measures and face masks. Focusing at the beginning only on some groups with transparent justification may help weaken the opposition to it.79

The high share of the population unwilling to vaccinate along with the number of people who are unable to receive the COVID-19 vaccine due to certain medical reasons suggests herd immunity may be out of reach. Information about the high death tolls and hospital overflows from the COVID-19 pandemic has recently flooded onto online media, but has apparently not convinced much of the worlds population to plan to be vaccinated. If the disturbing images being streamed live on social media cannot convince a fair share of the population to protect themselves from lethal risk, then educational or social campaigns may be limited in their effect. Educational efforts would be further undermined by the lack of trust in public authority figures, which fuels conspiracy theories and validates medical fake news. In this focused review we have discussed the challenges and opportunities faced during the management of COVID-19 in India.

Health-care systems across developed and developing nations are under tremendous pressure. The majority of this responsibility is being shouldered by frontline health-care workers to limit the spread of the novel coronavirus. They put their lives on the line in order to do so. Here we highlight some challenges faced by frontline HCW and propose certain recommendations to reduce the burden.

The exposure to the virus causes severe illness and mortality to a significant extent and also leads to physical and psychological exhaustion. This pandemic leads to health departments calling retired and experienced medical staff and clinical scientist back to work. Deficient supplies of personal protective equipment (PPEs) and other vital necessities is reported in various news channels all over the world. Majorly WHCs are affected and they are working in the emergency, they need PPEs and other vital necessities most.

In this pandemic, battling endless hours, staff shortages and deficient supplies, most are isolated from their families, affecting them physically, mentally, and emotionally, which will increase the morbidity and ill health.80 These mental health problems will not only affect decision making ability, judgement and attention of HCWs, but also affect the understanding the disease and have a long-lasting impact on their overall well-being.80

A few recommendations are proposed which are listed from all the information received around this issue.

Health-care staff/HCWs are also the most important resource as hospitals, equipments and PPEs in this pandemic situation. Post Traumatic Stress Disorder is reported in many health-care workers who have no time to protect themselves as well as their families. If any staff gets infected then they should be quarantined themselves, which leads to a shortage of staff and then healthy workers are stretched further for endless duties with lack of sleep and anxiety. For frontline health workers testing kits must be prioritized, as well as for weak communities (senior citizens) more susceptible to the virus and those who have many pre-existing diseases.81,82

Health-care workers face a high risk of getting infected as they take care of patients who are already infected. Protective clothing, sufficient hand sanitizers, washing paraphernalia and head covers are essential commodities which have to be provided to them in sufficient amount. Along with providing PPEs in adequate amount, its disposal methodology is also an important step across all the clinical areas since it can be one of the reasons of spreading infection.31

These are key phrases which provide the adequate time for the systems to gather resources and capacity to help in breaking the chain of transmission. The virus infects exponentially which is very clear and many will contract it very soon. State should provide premises to serve as isolation ward and quarantine spaces. All hospitals should use their full area to create control committees to monitor activities to ensure protocols are implemented for effective control. The loop has to be complete, involving community systems, governments and primary health-care workers are key, since not everyone will report to hospitals, if community transmission will be rampant.

The comfort and willingness in working for a health system which has an effective plan, magnifies many times in a pandemic. Protocols in local languages for better understanding and awareness material based on science research have been useful. Offering free transport service between work and home, childcare support and meal vouchers can reduce domestic stress and allow single-minded effort towards the health service.83

Apart from the various negative effect imposed by the pandemic, positive vibes of it cannot be neglected. The pandemic situation significantly improves air quality in different cities across the country, reduces GHGs emission, lessens water pollution and noise, and reduces the pressure on the tourist destinations, which may assist with the restoration of the ecological systems.84 These changes may be short term but are important for maintenance of environmental balance. Apart from this, various successful models like that of Dharavi and Kerala model were implemented which restricted the cases to a minimum through observing the spread in the localities, studying the prototype of spread, and strict use of methods to control the disease in Kerala. Dharavi restricted the coronavirus cases with a strategy of attack not defence and elucidated triumphant results in 2 months.85

There are more than 56 COVID-19 candidate vaccines in clinical evaluation of which 13 are in phase III trials and another 166 candidate vaccines are in preclinical evaluation (Table 1). All top candidate vaccines will be delivered through intra-muscular injection and are designed for a two-dose schedule.86 More recently our group has suggested the combinatorial use of childhood vaccines (BCG, MMR and OPV) along with the COVID-19 dedicated vaccines could be a potential strategy to control the COVID-19 pandemic worldwide.87

Table 1 Prospective Therapeutic Representative Against COVID-19 Disease

Strain B.1.1.7 was first detected in the United States in December 2020 followed by B.1.351, in South Africa P.1, in Brazil and Japan, B.1.427 and B.1.429. These two variants were first identified in California in February 2021. COVID-19 variant from India is B.1.617; one of the lineages is B.1.617.2, which has been detected most frequently in the US and the U.K.88 Recently the black fungus is now maiming COVID-19 patients in India. Mucormycosis is an invasive infection caused by a class of molds called mucormycetes. It has an overall mortality rate of 50%, and may be being triggered by the use of unhygienic oxygen cylinders and steroids, a life-saving treatment for severe and critically ill COVID-19 patients.89

In this review, we have been discussed the stories related to prevention strategies, chemotherapeutics and vaccines strategies to manage COVID-19. Apart from that we have discussed the challenges faced by HCWs and their prevention. Combating COVID-19 is still a challenge also due to the poorly-based counsel for using an experimental amalgamation of antimalarials and antimicrobials as treatment; the use of steroids; and antihypertensive drugs during the course of the disease. Interruption of the transmission of SARS-CoV-2 through engineered vaccines is top in the priority followed by the intense research to find out the potential treatment to control this viral infection.

All authors contributed to data analysis, drafting or revising the article, have agreed on the journal to which the article will be submitted, gave final approval of the version to be published, and agree to be accountable for all aspects of the work.

There is no funding to report.

Divakar Sharma and Dileep Tiwari were associated with Hericure Healthcare Pvt Ltd. Currently, Divakar Sharma is working in Maulana Azad Medical College at the time of this review. The authors reported no other potential conflicts of interest for this work.

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10. healthcare-in-europe.com [Internet]. Will resistant bacteria be the end of alcohol hand sanitizers? 2018. Available from: https://healthcare-in-europe.com/en/news/will-resistant-bacteria-be-the-end-of-alcohol-hand-sanitizers.html#. Accessed July 21, 2021.

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12. The Government of India issues simple guidelines, for controlling spread of COVID-19 in densely populated areas. Available from: https://pib.gov.in/PressReleseDetailm.aspx?PRID=1614064. Accessed July 21, 2021.

13. TIFAC explores best methods to revive Indian economy post COVID-19.Available from: https://dst.gov.in/tifac-explores-best-methods-revive-indian-economy-post-COVID-19. Accessed July 21, 2021.

14. TDB approves support for indigenous company for ramping up production of COVID-19 diagnostic kits.Available from: https://dst.gov.in/tdb-approves-support-indigenous-company-ramping-production-COVID-19-diagnostic-kits. Accessed July 21, 2021.

15. NBRI scientists develop herbal hand-sanitiser.Available from: https://vigyanprasar.gov.in/isw/NBRI-scientists-develop-herbal-hand-sanitiser.html. Accessed July 21, 2021.

16. Indian researchers to go for clinical trial of sepsis drug against novel coronavirus.Available from: https://vigyanprasar.gov.in/wp-content/uploads/Indian-researchers-to-go-for-clinical-trial-of-sepsis-drug-against-novel-coronavirus-21apr20.pdf. Accessed July 21, 2021.

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18. AMU alumnus develops COVID-19 testing kit, approved by ICMR. Available from: https://www.amu.ac.in/about3.jsp?did=2495. Accessed July 21, 2021.

19. Bione launches rapid COVID-19 at-home screening test kit after ICMR approval.Available from: https://zeenews.india.com/india/bione-launches-rapid-COVID-19-at-home-screening-test-kit-after-icmr-approval-2273752.html. Accessed July 21, 2021.

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Challenges and Successes of Dealing with COVID-19 in India | RRTM - Dove Medical Press