Reactor Design for Loading of Paclitaxel into PLGA Nanoparticles

Team Members: Matthew Pickering, Andrea Kuliasha, Lauren Wills, Sasha Thomasson,
Gavin Vess, Jane Gong
Advisors: Dr. Chenming Zhang, Dr. Veronica Mbaneme-Smith, Dr. Matt Dickson
Instructor: Dr. Cully Hession

TABLE OF CONTENTS
TABLE OF CONTENTS..................................................................................................................................... 1
ABSTRACT...................................................................................................................................................... 2
DESCRIPTION OF CLINICAL NEED AND PROBLEM ......................................................................................... 2
PROBLEM OBJECTIVE STATEMENT ............................................................................................................... 3
DOCUMENTATION OF DESIGN AND PROTOTYPE ......................................................................................... 3
PROTOTYPE OF THE FINAL DESIGN ............................................................................................................... 4
FUNCTIONALITY AND PROOF THAT THE DESIGN WILL SOLVE THE PROBLEM ............................................. 6
PATENTABILITY.............................................................................................................................................. 7
ANTICIPATED REGULATORY PATHWAY......................................................................................................... 8
REIMBURSEMENT ......................................................................................................................................... 9
ESTIMATED MANUFACTURING COST ......................................................................................................... 10
POTENTIAL MARKET AND IMPACT .............................................................................................................. 10
REFERENCES ................................................................................................................................................ 12
APPENDIX .................................................................................................................................................... 14

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ABSTRACT

Nanoparticles have increasingly been used as an alternative to traditional

chemotherapy, in part because of their ability to alleviate some of the side effects experienced
by patients who receive free drug chemotherapeutics. Conventional methods for drug
encapsulation into nanoparticles often prove too expensive and unreliable for industrial-scale
production. This senior design team has designed an industrial-scale process that alleviates
those concerns by providing robust results, reduced waste, and adjustable throughput. Using
AutoCAD 3D (Autodesk, Mill Valley, CA), the team designed a coaxial turbulent jet mixer, based
on the design implemented by Lim et al. (2014). Two streams of water at equal flow rates
converge in a T-union and then flow together into one pipe. Inserted through this pipe is a blunt
needle which carries the “organic phase” containing Paclitaxel and poly(lactic-co-glycolic acid)
(PLGA) dissolved in acetone. The two phases combine downstream, leading to formation of
PLGA-encapsulated Paclitaxel. Economic analysis of the process was performed to determine a
hypothetical yearly production rate based on patient demand. Computational Fluid Dynamics
(CFD; ANSYS, Inc., Canonsburg, PA) allowed for flow rate analysis to determine the
appropriate volumetric flow rates necessary to accomplish the desired rate of product
production. SuperPro Designer (Intelligen, Inc., Scotch Plains, NJ) was used to design the
overall biomanufacturing of the value-added chemotherapeutic based on implementation of the
coaxial turbulent jet mixer designed by Lim et al. (2014) to demonstrate necessary downstream
processes for purification, preparation of finished product, and evaluation of design profitability.

DESCRIPTION OF CLINICAL NEED AND PROBLEM

Nanomedicine has a bright future in the pharmaceutical and biotechnology industries,

especially due to its ability to deliver drugs that are not soluble in water and to alleviate some of
the toxicity that is often associated with administration of free drugs, especially in the context of
cancer treatment (Langer, Omid C. & Langer, Robert, 2009). One major challenge in the
pharmaceutical industry pertaining to nanoparticles is to reduce the amounts of drug and energy
wasted in drug-loaded nanoparticle formation. The volume of waste can significantly increase
the cost of the finished drug product (Govender, Stolnik, Garnett, Illum, and Davis, 1999).
Reducing wasted energy, drugs, nanoparticles, and solvents can lower the cost of the finished
drug product, which will allow a pharmaceutical company to sell their products at a lower price
without sacrificing their profit margin. This price reduction would also benefit the insurance
company and patient, contributing a solution to address the concerns of rising healthcare
burdens in the United States. Increasing the yield of final product also minimizes the waste of
drugs and nanoparticles (Luo, Okubo, Nangrejo, and Edirisinghe, 2017). The number of
processing steps can be reduced, lowering labor costs because fewer unit operators will be
required, which in turn would lower manufacturing costs (Luo et al., 2017; Turton, Bailie,
Whiting, and Shaelwitz, 2009). The group plans to evaluate different reaction processes for
nanoparticle formation, the materials used for construction of the reactor, and the conditions (ex:
pH, temperature) within the reactor in order to reduce these costs.

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 To garner a pharmaceutical company’s attention and interest, a design process must be
reproducible and fairly easy to scale up (Albisa, Piacentini, Sebastian, Arruebo, Santamaria,
and Giorno, 2017). Reproducibility is an important criteria to the group, leading to the selection
of PLGA out of all nanoparticles reviewed, which has been shown to be reliable in efficient drug
loading (Schubert, Delaney Jr., and Schubert, 2011). When evaluating various fabrication
methods, the group will take into account the properties of PLGA, the properties of the drug, and
the properties of the nanoparticle-encapsulated drug. In addition, the group will look at the
capability of each design to follow standards that include manufacturing guidelines, workers’
safety, pharmaceutical requirements, and environmental considerations.

PROBLEM OBJECTIVE STATEMENT

To address the concerns outlined above, the team has designed an industrial-
scale production process for the loading of Paclitaxel into PLGA nanoparticles. Certain unit
operations included in this process have been evaluated and characterized on a small scale, but
this design process is the first of its kind to the best of the team’s knowledge. The team has
focused on optimizing the various unit operations of the process to achieve minimum
nanoparticle polydispersity, minimize overall costs, and maximize encapsulation efficiency. At
the core of the team’s industrial process is a microfluidic coaxial jet mixer design and optimized
by Lim et al. (2014) for small-scale production of drug-encapsulated nanoparticles. Lim et al.
(2014) allude to potential application of this device to full-scale production of encapsulated
drugs. As a part of this project, this design was implemented in SuperPro Designer as the
reaction step. To further revise this process, experiments would need to be performed that verify
the flow rates previously characterized by Lim et al. (2014) in the modified device used here.
While alternative solvents and unit operations were previously explored by the team, further
optimization of each unit operation could potentially yield a more efficient and economically
favorable final project.

DOCUMENTATION OF DESIGN AND PROTOTYPE

Using Inventor software, a 3D rendering of the coaxial turbulent jet mixer was created
based on the design and dimensions detailed by Lim et al. (2014). The design consists of three
pipes intersecting at a T-union and a blunt needle entering through the backside of the
aforementioned union. This simple assembly is intentional as it keeps the technology accessible
and doesn’t restrict its use to specialized microfluidic labs. This system is regulated through two
parameters: the Reynolds number and the flow velocity ratio between the inner and outer tubes.
Such tight control over the mixing environment allows for a more consistent product in terms of
nanoparticle size and chemical characteristics (Lim et al., 2014). The continuous nature of this
system also reduces batch-to-batch variability. Scale-up of batch processes often leads to
additional work in terms of re-optimizing the system, typically accomplished through a trial and
error process (Lim et al., 2014). This jet mixer, however, is able to reach a production rate of
3.15 kg/day, which is well over the 1 kg/day needed to be classified as an industrial scale
process (Lim et al., 2014). Therefore, this system does not require any dimensional change to

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increase the output of the system. The jet mixer is constructed out of PTFE-lined stainless steel
to provide mechanical strength, while also being solvent compatible.
The SuperPro design deliverable for this project encompasses all the unit operations
necessary to convert the raw materials into a drug-loaded PLGA nanoparticle with potential to
serve as a chemotherapeutic agent for cancer patients. The process begins with a blending tank
in which PLGA and its solvent, acetone, are mixed with the free drug. From this point, the
solution moves through the jet mixer, where the solution precipitates out into loaded drug
product, in addition to some remaining unloaded polymer and free drug. After this reaction, the
solution is flowed through an ultrafiltration/diafiltration unit to recover the product and un-loaded
polymer. Free drug in solution passes through this process and is sent to a reverse osmosis unit
where the free drug can be recovered. Next, the product and remaining components of polymer
are passed through a size exclusion chromatography gel column. During this unit operation, the
gel column, which is filled with beads with a specified pore size, allows for the components in
solution to flow through at various rates based on molecular weight. Components with molecular
weights below the pore size will take longer to flow through the column while larger
components, here the product, will pass through more quickly. The product will then be passed
through a disk stack centrifuge to reduce liquid content. Finally, the product will be freeze dried
with 1% sucrose as recommended by Holzer et al. (2009) for stabilization to allow for packaging
and labeling, ideally being reconstituted in solution upon arrival at a patient care facility. Cycling
and unit operation staggering have been applied in the software to maximize productivity of the
process.
The ANSYS Computational Fluid Dynamics (CFD) design deliverable models how fluid
will flow through the jet mixer. The AutoCAD design deliverable was adapted to be used for the
CFD model, as exemplified in appendix X figures 1 and 2. The first step in preparing the CFD
model was establishing the mesh fluid domain. The size of the reactor Mesh element sizes
considered were 0.3, 0.1, and 0.05. A mesh size of 0.1 provided the lowest mixing time of
14.905 ms, so that is the size used for further simulations. Figure 3 provides a visual of the
mixer when the mesh condition was applied while figure 4 provides a close up view of one end
of the mixer. The next consideration for setting up the model is constructing the fluid
parameters, including material and flow rate specifications. Inlet flow rates and Reynolds
numbers were calculated based on the lab-scale model of the design found in [the ctjm paper].
Using the inlet information, outlet data was calculated. Certain parameters, like outlet density
and viscosity, were calculated based on the assumption that the outlet would consist of 1/11 of
acetone and 10/11 of water. In the first recorded simulation, the model exemplified basic flow
rate parameters based on water entering through the two inlet pipes with a Reynolds number of
1635 and a flow rate of 0.459 m/s, while acetone entered through the needle with a Reynolds
number of 6839 and a flow rate of 8.175 m/s. Using data from the fluid dynamic model, mixing
length and mixing time were calculated. (Appendix y, Figure y).

PROTOTYPE OF THE FINAL DESIGN

AutoCAD Drawing of our coaxial turbulent jet mixer

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Our ANSYS CFD Simulation:

https://drive.google.com/file/d/0BxIf6lps3WUsVG5PMVljeGZfVy1wTE5TanpOeDdadkt6ekpN/view?us
p=sharing
SuperPro Drawing of Jet Mixer and Downstream Processes:

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FUNCTIONALITY AND PROOF THAT THE DESIGN WILL SOLVE THE PROBLEM

One objective of this design is to achieve specific physicochemical properties of the

product, which will impact how it performs in the body. Lim et al. (2014) was able to achieve a
relatively consistent size for PLGA nanoparticles, comparably smaller than those produced from
a batch system. In addition, they exhibited the ability to control the size of the nanoparticles by
increasing the Reynolds Number.
In order to form a basis for comparison, coaxial turbulent jet mixing was compared with
one of the best methods currently used in industry--nanoprecipitation. In terms of design
scalability, the higher rate of turbulence needed for scaled-up nanoprecipitation systems
generates increased size variability amongst products, which often result in undesirable larger
particle sizes (Rivas et al., 2017). With regards to productivity, a jet mixing design could
significantly surpass current production rates for nanoparticle-encapsulated drugs using other
methods, such as double emulsification and nanoprecipitation, as a result of the continuous
nature of the process (Lim et al., 2014). Obtaining replacement parts will be much easier for
nanoprecipitation due to longevity of use when compared to jet mixing; however,
nanoprecipitation is not always run continuously, which is the mode of operation for the jet mixer.
A continuous operating mode reduces input and product losses and decreases labor costs related
to cleaning (Rivas et al., 2017). As a result of fabrication simplicity when designing the coaxial

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turbulent jet mixer, easily replaceable high quality parts are utilized to reduce natural depreciation
(Lim et al., 2014). In addition, coaxial turbulent jet mixers need very little cleaning because high
flow rates wash away precipitates, which prevent buildup in the walls of the system even after
long-term use (Lim et al., 2014). Based on the previously mentioned advantages of using a coaxial
turbulent jet mixing design, our design seems to be a more favorable approach than the currently
utilized industrial standard, nanoprecipitation.
In order to further validate our design, the ANSYS CFD model was utilized to find the
mixing length and time of the coaxial turbulent jet mixer. The mixing length was found as defined
by Lim et al. as the distance from the needle tip at which 90% of the final end-of-stream water
mass fraction is reached in the center of the streams flow (2014). From this length, the average
velocity of the outlet pipe could be used to calculate the overall mixing time using the simple
definition of velocity as length over time. These results yielded a similar decreasing trend in mixing
time with higher Reynolds numbers to those shown by Lim at al. as shown in Figure 10, verifying
our model with experimentally derived values (2014). The system modeled is slightly different
than the experimental system since acetonitrile is used instead of acetone in their experiments.
However, the overall trend is assumed to hold for both systems as validated by our model. This
leads to the best derived mixing time to be 15.04 milliseconds when a Reynold’s number of 3000
is utilized. This result can then be translated into the SuperPro model for better economic analysis
of the system.

Figure 10: Mixing time vs. Reynold’s number for coaxial turbulent jet mixer as modeled using CFD

PATENTABILITY

Patentability will be discussed in two parts, the coaxial turbulent jet mixer and the
finished drug product PLGA encapsulated Paclitaxel. It would be almost impossible to patent
use of a coaxial turbulent jet mixer for two reasons. The first is that someone has a provisional

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patent application along with a pending nonprovisional patent application in the United States
(Lim et al., 2015), and there is also a patent application with the World Intellectual Property
Organization (WO) (Lim et al., 2015). Even if the patents were never filed, the same authors
also published an article describing the technology on May 13, 2014 (Lim et al., 2014). Since
this technology has been described in literature for over a year, a patent application for the
coaxial turbulent jet mixer would be denied in the United States because of failure to meet the
requirement of being novel (Whitehead, 2018). If the nonprovisional patent application
US14523869 is granted approval, one would have to pay a royalty in order to construct it, buy
the product, or risk being sued. If a patent could be filed, it would fall under the category of a
utility patent and receive 20 years of protection in the US (Whitehead, 2018). Other methods
that are commonly used in industry for nanoparticle formulation are nanoprecipitation and
emulsification (Wang et al., 2016). Patents have been successfully obtained for a method
similar to the use of nanoprecipitation (Fessi et al., 1992) and emulsification (Vanderhoff et al.,
1979) methods; however, these are not overarching for all nanoprecipitation and emulsification
methods. Most of the patents that have been successful in regards to nanoparticle formulation
are specific to a particular nanoparticle or branch of nanoparticles.
A patent for PLGA encapsulated Paclitaxel for the treatment of cancer would also be
difficult to obtain because the product of interest is well known for cancer treatment (Tuesca,
2018) and has been described in prior literature (Danhier et al., 2012). In order to get a patent
approved, a novel surface modification of the nanoparticle would be the best avenue. This
would be favorable regardless of patentability because PLGA drug release has been shown to
be favorable with surface modifications such as PEGylation as opposed to with no modifications
(Danhier et al., 2009). A less likely, secondary approach to obtaining a patent would be to utilize
the drug for treatment of an entirely different disease or form of cancer from previously
indicated. Paclitaxel has been approved for ovarian cancer, breast cancer, non-small cell lung
cancer, small cell lung cancer, AIDS related Kaposi Sarcoma, and several other cancers
(Drugs.com, 2018), so the likelihood of achieving a patent through that avenue would be
unlikely. If a patent could be obtained for the drug, it would also fall under the category of a
utility patent and receive 20 years of protection in the United States (Whitehead, 2018). There is
one nanoparticle drug formulation with Paclitaxel that has been patented and approved by the
FDA in 2005 known as Abraxane (Abraxis Bioscience, 2006). Some of the initial challenges to
our drug if approved would include the drugs Avastin, Herceptin, Alimta, Xeloda, Tarceva,
Afinitor, Erbitux and Zoladex (Stone, 2018). Although some of these drugs such as Herceptin
will soon lose their exclusivity (Philippidis, 2018), it is anticipated that because the cancer
therapy market is growing and so extremely lucrative additional competitors will saturate the
market in future years (Tatkare, 2013).

ANTICIPATED REGULATORY PATHWAY

The three distinct regulatory pathways for drug approval are the 505 (b) (1) for
new drugs, 505 (j) for generics, and 505 (b) (2) for hybrids. Because Paclitaxel is not a new drug
the 505 (b) (1) pathway would not be an option. The 505 (j) pathway would also not be an option
because there is no PLGA encapsulated Paclitaxel out on the market. Even if it was possible, to
go through extensive clinical trials before generating a generic would not be an economically
prudent decision. Thus, the 505 (b) (2) is the preferred pathway in this situation. This pathway
would allow the manufacturer to cite information from prior studies done on the different

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formulations using the Active Pharmaceutical Ingredient (API) Paclitaxel (Camargo, 2018;
Chandanais, 2017). To date, there are 15 FDA approved Paclitaxel drug products, Taxol being
the original, 13 generics, and one drug that went through the 505 (b) (2) pathway, Abraxane
(FDA, 2018a). Even with citing the prior studies, our product would still likely have to undergo
clinical trials to attain approval. Before beginning clinical trials, the sponsor would have to file an
Investigational New Drug (IND) Application (FDA, 2016a). In this application, the sponsor has to
provide or cite the evidence that the drug is reasonably safe from animal testing, information on
the composition of the drug and how it is manufactured, knowledge about the manufacturer
themselves, detailed protocol for how the drug will be tested, agreements to obtain informed
consent from research subjects and submit to a review board, all while abiding by IND
regulations set forth in title 21 part 312 of the code of federal regulations (FDA, 2016a). Clinical
trials can begin in 30 days after the FDA receives the IND unless the FDA decides to approve
the sponsor earlier or does not approve the IND and places it on clinical hold. Clinical trials
typically take place in three phases before market approval (FDA, 2018b). The first phase takes
several months of testing to determine a safe dosage. The second phase tests for efficacy and
safety, and this can take up to 2 years. The third phase, which takes about 1 to 4 years,
continues testing for efficacy and monitoring for adverse reactions. If clinical trials are
successful, the sponsor would then submit a New Drug Application (NDA) to the FDA’s Center
for Drug Evaluation and Research (FDA, 2016a). This information would include the previous
information of the IND Application along with the results from human clinical trials and the
labeling (FDA, 2016a). After the NDA is submitted, it will receive either a priority or standard
designation (FDA, 2013). The priority designation means that the goal for approval of the drug
would be 6 months after submission of the NDA as opposed to the standard 10 months (FDA,
2013). The criteria for receiving a priority designation is that the drug would improve the safety
or efficacy of treating a serious or life threatening illness or condition as defined by part 312.81
under title 21 of the code of federal regulations (FDA, 2013). Since non-small cell lung cancer
and breast cancer are both life threatening, under the assumption that our drug improves the
treatment it would receive a priority designation. While the goal for NDA approval is 6 months
after submission, the expected approval timeline would be expected to be around 8 months,
which is still an improvement compared to the standard approval timeline of 10.1 months (FDA,
2016b). In total, the pathway to approval is expected to take approximately 8 years for
completion, which is the typical timeline for approval for cancer treatments (University of
Arizona, 2018).

REIMBURSEMENT

It is expected that poly(lactic-co-glycolic acid)-bound Paclitaxel will be covered by

Medicare/Medicaid and other insurance plans. The nanoparticle-drug product resulting from the
design is comparable to the FDA-approved Abraxane. Abraxane is Paclitaxel albumin-bound
particles. Like the PLGA-Paclitaxel product, it is an injectable Paclitaxel containing nanoparticle.
Abraxane treats metastatic breast cancer, metastatic non-small cell lung cancer and metastatic
adenocarcinoma, which is similar in use to our product. A patient with a diagnosis of one of the
previous disease will be eligible to receive Abraxane and it will be covered by
Medicare/Medicaid. In addition, most private healthcare companies will also cover Abraxane.
Celgene Support will has a support line where a member of their team will perform a review of

9
ones insurance plan and help identify available options for the patient. There are options if one
does not have insurance that covers Abraxane. Firstly, if insurance is denied there is an appeal
process that one can undergo. If someone does not have health insurance, there are programs
that can help provide financial support. For example, Celgene, the company behind Abraxane,
offers a Celgene Patient Assistance Program which provides aid for those who financially
qualify to cover the cost of the drug. Overall, one would expect that because our drug product is
of similar caliber, that similar reimbursement and coverage policies would be in place to make
this product available to everyone. The PLGA-Paclitaxel product will be covered by
Medicare/Medicaid and programs will be available to help users maneuver through the
insurance process.

ESTIMATED MANUFACTURING COST

For the coaxial turbulent jet mixer itself, the blunt needle is a 23 G blunt needle
that comes in packages of 100 needles at a time for $71.20 from Sigma-Aldrich. The needle is
treated as a consumable and is replaced every 24 cycles, amounting to a cost of $187 per year.
The cost of the T Union provided by Plasmatech is $29.85 per unit(). The pumps, controls, and
software are provided by Cetoni for 10,000 Euros or $12,000 (), and two controls will be used to
measure the flow from the outlet pipe of the jet mixer each cost 500 euros or $600 each.
For raw materials, the process will use PLGA, Paclitaxel, Acetone, Deionized water,
water for injection, and sucrose. Phyton Biotech, a large-scale provider of Paclitaxel, sells the
drug for $50.00 per gram. Acetone can be found at a market value of $0.73 per kilogram (S&P
Global Platts, 2015). Medical grade PLGA can be bought at a market price of $10 per gram
(Doctor Anthony Tuesca, personal communication, 4 April 2018). DI water used prior to the
UF/DF unit operation is typically $0.001 per kg in bulk (). The team used the SuperPro default
cost of $0.10 per kg for water for injection (). The SuperPro default cost of $0.80 per kg was
also used for sucrose (). Additional consumables used in the process include the Planova 15N
Ultrafiltration membrane (PPV specification), Sephacryl S-400 resin for gel filtration, glass
ampoules, drug labels, and rigid boxes. Sephacryl S-400 resin can be obtained for $664/L (GE
Healthcare). The cost of the Planova 15N membrane was not obtained, so the cost was
estimated by a linear system of equations developed from SuperPro’s default values based on
pore size (See appendix). The cost of the membrane was estimated to be $973.20 per m^2.
Thomas Scientific provides 1 mL glass ampoules in packages of 1152 ampoules for $369.11, or
$0.3204 per ampoule. When buying in bulk from Medi-Dose, drug labels are $0.0396 per label.
No quote was obtained for boxes, but the cost was estimated to be $5.00 a box (). That does
not include the cost of linear, brochures, box labels, and other elements that typically go into
packaging with drugs ().

POTENTIAL MARKET AND IMPACT

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 The oncology market is rapidly growing. As of 2016 the value of the oncology
market was around $82 billion (AstraZeneca, 2016), and in 2020 the market size is projected to
increase to over $111 billion (Tatkare, 2013). The initial indications of our drug product would
include the most common forms of cancer, which are breast cancer and non small cell lung
cancer (WHO, 2018), and potentially future targeting for pancreatic cancer and ovarian cancer,
as well as AIDS related Kaposi Sarcoma. There are expected to be over 268,670 new cases of
breast cancer and 234,030 new cases of lung and bronchus cancer in the United States in 2018
(American Cancer Society, 2018). In 2012, the World Health Organization estimated that lung
and breast cancer affected approximately 3.5 million new people each year (WHO, 2018), and
those estimates are expected to grow significantly over the course of the 21st century (World
Cancer Research Fund International, 2018).
The number of potential patients is not expected to be the limiting factor in determining
market potential; instead, the limits of sales will be set by access, cost, efficacy, and safety in
opposition to competing drugs. Access and cost are expected to be the biggest limiting factors
in product sales, particularly when looking internationally. PLGA encapsulated Paclitaxel
requires intravenous administration, which is unfortunately very limited or too expensive in less
developed countries (Tuesca, 2018). This would eliminate approximately 52.83% of potential
patients worldwide for breast cancer and 58.45% of potential patients worldwide for lung cancer,
which even when accounting for the projected increase between 2012 to 2030 of 50% (World
Cancer Research Fund International, 2018) gives an estimation of absolute maximum potential
consumer base of 1,804,185 new cases per year (see appendix). In terms of a more realistic
estimate, our goal would be to make our formulation less expensive than the most similar
formulation with Paclitaxel as the API, Abraxane (Abraxis Bioscience, 2006), with improved
efficacy and safety of the product. Otherwise due to the increasing negative public image and
medical community perceptions of Abraxane not being effective enough in treatment to justify
the cost (Beasley, 2015), it would be difficult to market our drug and achieve profitability.
Pharmaceutical companies typically do not sell to the end consumer which is the patient
who needs chemotherapy. In many cases the insurance company that pays a significant portion
of treatment costs (Mattingly, 2012). Therefore, the drug is often sold either to a hospital or to a
wholesale distributor, who then in turn sells the product to the hospital where the patient is
treated (Mattingly, 2012). The drug will be sold to wholesale distributors who will then extend the
reach of our drug, which we intend to sell for a price of $2.5 per mg.

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REFERENCES

Abraxis Bioscience. (2006). Abraxane: Oncologic drugs advisory committee meeting.

American Cancer Society. 2016. What is small cell lung cancer? American Cancer Society. Retrieved
from https://www.cancer.org/cancer/small-cell-lung-cancer/about/what-is-small-cell-lung-cancer.html.
American Cancer Society. (2018). Cancer facts & figures 2018. American Cancer Society. Retrieved
from https://www.cancer.org/content/dam/cancer-org/research/cancer-facts-and-statistics/annual-cancer-
facts-and-figures/2018/cancer-facts-and-figures-2018.pdf
AstraZeneca. (2016). Astrazeneca annual report and form 20-f information 2016. AstraZeneca. Retrieved
from https://www.astrazeneca.com/investor-relations/annual-reports/annual-report-2016.html
Beasley, D. 2015. U.S. cancer doctors drop pricey drugs with little or no effect. Los Angeles, California:
Reuters. Retrieved from https://www.reuters.com/article/us-usa-healthcare-cancer-insight/u-s-cancer-
doctors-drop-pricey-drugs-with-little-or-no-effect-idUSKCN0S20DG20151008.
Camargo. (2018). What is 505 (b) (2)? . Cincinnati, OH: Camargo. Available at:
https://camargopharma.com/what-is-505b2/.
Chandanais, R. (2017). 505 (b)(2) regulatory pathway for new drug approvals. Pharmacy Times.
Danhier, F., Lecouturier, N., Vroman, B., Jérôme, C., Marchand-Brynaert, J., Feron, O., & Préat, V.
(2009). Paclitaxel-loaded pegylated plga-based nanoparticles: In vitro and in vivo evaluation. Journal of
Controlled Release 133(1):11-17.
Danhier, F., Ansorena, E., Silva, J. M., Coco, R., Le Breton, A., & Préat, V. (2012). PLGA-based
nanoparticles: An overview of biomedical applications. Journal of Controlled Release 161(2):505-522.
Desai N. Challenges in Development of Nanoparticle-Based Therapeutics. The AAPS Journal.
2012;14(2):282-295. doi:10.1208/s12248-012-9339-4.
Drugs.com. 2018. Paclitaxel. Available at: https://www.drugs.com/monograph/paclitaxel.html.
FDA. 2013. Review designation policy: Priority (p) and standard (s). O. o. N. Drugs, ed. Silver Spring,
MD: FDA.
FDA. 2016a. New drug development and review process. Silver Spring, Maryland: FDA. Retrieved from
https://wayback.archive-
it.org/7993/20170723164036/https://www.fda.gov/Drugs/DevelopmentApprovalProcess/SmallBusinessA
ssistance/ucm053131.htm.
FDA. 2016b. New drug applications (nda) & biologic license applications (bla) approval times calendar
years 1993-2016. Silver Spring, MD: FDA.
FDA. 2018a. Drugs@fda: Fda approved drug products. Silver Spring, MD: FDA. Available at:
https://www.accessdata.fda.gov/scripts/cder/daf/
FDA. 2018b. Step 3: Clinical research. Silver Spring, MD: FDA. Available at:
https://www.fda.gov/ForPatients/Approvals/Drugs/ucm405622.htm.

12
Fessi, C., Devissaguet, J.-P., Puisieux, F., & Thies, C. (1992). Process for the preparation of dispersible
colloidal systems of a substance in the form of nanoparticles. U.S. Patent No. 5118528A.
Govender, T., Stolnik, S., Garnett, M. C., Illum, L., & Davis, S. S. (1999). Plga nanoparticles prepared by
nanoprecipitation: Drug loading and release studies of a water soluble drug. J. of Controlled Release
57(2):171-185.
Gunderson, J. 2018. Water treatment, chemical and pharmaceutical industries. Tulsa, OK: PennWell
Corporation. Retrieved from http://www.waterworld.com/articles/iww/print/volume-12/issue-05/feature-
editorial/water-treatment-chemical-and-pharmaceutical-industries.html.
Helms, M. 2018. GE Healthcare Life Sciences Virginia Tech Project. M. Pickering, ed. Philadelphia, PA:
GE Healthcare Life Sciences.
Holzer, M., et al. (2009). "Physico-chemical characterisation of PLGA nanoparticles after freeze-drying
and storage." European Journal of Pharmaceutics and Biopharmaceutics 72(2): 428-437.
International, W. C. R. F. 2018. Worlwide data. Available at: https://www.wcrf.org/int/cancer-facts-
figures/worldwide-data.
Lim, J.-M., Gilson, L. M., Chopra, S., Farokhzad, O. C., Karnik, R., & Swami, A. (2015). High-
throughput synthesis of nanoparticles. U.S. Patent No. 2,015,017,454,9A1.
Lim, J.-M., Swami, A., Gilson, L. M., Chopra, S., Choi, S., Wu, J., Langer, R., Karnik, R., & Farokhzad,
O. C. (2014). Ultra-high throughput synthesis of nanoparticles with homogeneous size distribution using
a coaxial turbulent jet mixer. ACS Nano 8(6):6056-6065.
Lince, F., Bolognesi, S., Stella, B., Marchisio, D. L., & Dosio, F.(2011). Preparation of polymer
nanoparticles loaded with doxorubicin for controlled drug delivery. Chem. Eng. Res. and Des.
89(11):2410-2419.
Lichtman S. M., Hurria A., Cirrincione C. T., Seidman A. D., Winer E., Hudis C., Cohen H. J., and Muss
H. B. (2012). "Paclitaxel efficacy and toxicity in older women with metastatic breast cancer: combined
analysis of CALGB 9342 and 9840." Ann Oncol 23(3): 632-638.
Mattingly, J. 2012. Understanding Drug Pricing. US Pharmacist. Retrieved from
https://www.uspharmacist.com/article/understanding-drug-pricing.
Pharmate Systems. 2018. USP Standards for Pharmaceutical Water. Pharmatesystems.com. Retrieved
from http://www.pharmatesystems.com/usp-standards-for-purifed-water/.
Philippidis, A. 2018. Top 10 best-selling cancer drugs, Q1–Q3 2017. Genetic Engineering &
Biotechnology News. Retrieved from https://www.genengnews.com/the-lists/top-10-best-selling-cancer-
drugs-q1q3-2017/77901033.
Platts, S. P. G. 2015. Acetone: European spot price rise; us export pricing stable; asian price unchanged.
S&P Global.
Rivas, C. J. M., Tarhini, M., Badri, W., Miladi, K., Greige-Gerges, H., Nazari, Q. A., Rodríguez, S. A. G.,
Román, R. Á., Fessi, H., & Elaissari, A. (2017). Nanoprecipitation process, from encapsulation to drug
delivery. Intl. J. of Pharm. 532(1):66-81.

13
Stone, K. 2018. Top 20 blockbuster cancer drugs. The Balance. Available at:
https://www.thebalance.com/top-cancer-drugs-2663234.
Tatkare, D. 2013. Oncology drugs market overview. Allied Market Research. Portland, OR.
Tuesca, A. 2018. Team nanoparticles progress update. M. Pickering, A. Kuliasha, L. Wills, J. Gong, G.
Vess, S. Thomasson, and M. Dickson, eds. Gaithersburg, Maryland.
University of Arizona, 2018. The university of arizona cancer center. Tuscon, Arizona: University of
Arizona, National Cancer Institute. Retrieved from http://uacc.arizona.edu/patients/clinical-trials/faq.
University of Sevilla. 2018. Chapter 3. Activated Carbon Columns Plant Design. Sevilla, Spain: Library
of Engineering. Retrieved from http://bibing.us.es/proyectos/abreproy/20087/fichero/CHAPTER+3.pdf.
US Pharmacopeia.2018.Water for pharmaceutical purposes. Rockville, MD: US Pharmacopeia.
Vanderhoff, J. W., El-Aasser, M. S., & Ugelstad, J. 1979. Polymer emulsification process. U.S. Patent
No. 4177177A.
Wang, Y., Li, P., Truong-Dinh Tran, T., Zhang, J., & Kong, L. (2016). Manufacturing techniques and
surface engineering of polymer based nanoparticles for targeted drug delivery to cancer. Nanomaterials,
6(2):26.
Whitehead, R. 2018. Intellectual property & technology transfer. Blacksburg, VA: Virginia Tech
Intellectual Properties, Inc.
WHO. 2018. Cancer. Available at: http://www.who.int/mediacentre/factsheets/fs297/en/.

APPENDIX

Figures

Figure 1

14
Figure 2

Figure 3

15
Figure 4

Figure 5

16
Figure 6

Figure 7

17
Figure 8

Figure 9

18
Calculations:
Equations for UF/DF
Pore size = 25 nm
Cost…
Dft membrane: 0.45 μm = 450 nm
Dft DEF cartridge: 0.2 μm = 200 nm
Dft R0 membrane: 0.45 μm = 450 nm
MF membrane Biotech: 0.45 μm = 450 nm
UF membrane Biotech: 0.005 μm = 5 nm

Unit Cost
Dft membrane 0.45 μm = 450 nm $40,000/m2
Dft DEF cartridge 0.2 μm = 200 nm $1,000/m2
Dft R0 membrane 0.45 μm = 450 nm $15/m2
MF membrane Biotech 0.45 μm = 450 nm $735.84/m2
UF membrane Biotech 0.005 μm = 5 nm $981.11/m2

Cost Calculations Example

𝑦 = 𝑎𝑥 + 𝑏
$981.11
= 𝑎(5 𝑛𝑚) + 𝑏
𝑚2
$735.84
= 𝑎(450 𝑛𝑚) + 𝑏
𝑚2
Using System of Equations:
$245.27
= −445𝑎
𝑚2
𝑎 = −$0.5512/(𝑚2 × 𝑛𝑚 𝑝𝑜𝑟𝑒 𝑠𝑖𝑧𝑒)
$981.11
= −$0.5512(5 𝑛𝑚) + 𝑏
𝑚2
Rejection is determined by Zeman and Wales equation…
𝑑𝑖
𝜆=
𝑑𝑝
Where di = species diameter and dp = pore size.
Pore size is given as 25 nm. From this value we must calculate the diameter of the product
components…

19
 1. Water → not need, zero Ri
2. Acetone → not needed, zero Ri
3. Paclitaxel (free drug)
4. PLGA-encapsulated Paclitaxel
For rejection of free drug…
Paclitaxel:
Molecular weight: 853.906 g/mol
ρ = 1.40 g/cm3
853.906 𝑔 𝑐𝑚3 𝑚𝑜𝑙 1.0128 × 10−21 𝑐𝑚3 (10𝑛𝑚7 )3
× × = ×
𝑚𝑜𝑙 1.4 𝑔 6.02214 × 1023 𝑚𝑜𝑙𝑒𝑐𝑢𝑙𝑒𝑠 𝑚𝑜𝑙𝑒𝑐𝑢𝑙𝑒 (1 𝑐𝑚)3

= 1.0128173 𝑛𝑚3 𝑝𝑒𝑟 𝑚𝑜𝑙𝑒𝑐𝑢𝑙𝑒

4 𝑑 3
3
1.0128 𝑛𝑚 = 𝜋 ( )
3 2
𝑑 = 1.245979315 𝑛𝑚
𝑑𝑃𝑇𝑋 1.245 𝑛𝑚
𝜆= = = 0.0498 < 0.5
𝑑𝑃 25 𝑛𝑚
𝑅𝑖 = (𝜆(2 − 𝜆))2 = 0.0094467518
For PLGA-encapsulated Paclitaxel:
𝑑𝑖
- If the pores were strictly uniform, then we would say ≥ 1, 𝑠𝑜 𝑅𝑖 = 1. But some
𝑑𝑝
variability exists, so we will apply a pessimistic assumption. Assume pore size
distribution is 25 ± 5 𝑛𝑚 so that the max pore size is 30 nm. Our nanoparticle size
distribution is 25 to 60 nm. Using a triangle distribution with a height of one (y axis is
unitless) and a width of 60nm – 25nm = 35. This yields a triangle area of 17.5. To
determine the area for which we would apply the Zeman and Wales equation, we used a
height of 1 and a width of 30nm – 25nm = 5, and this gives us an area of 2.5.
2.5
Ratio of nanoparticle that may be rejected: 17.5 = 0.1429

1 − 0.1429 = 0.8571
𝑂𝑣𝑒𝑟𝑎𝑙𝑙 𝑟𝑒𝑗𝑒𝑐𝑡𝑖𝑜𝑛 = (𝑅𝑖 )(0.1429) + (1)(0.8571)
Where Ri is 25 to 30 nm. Ri is no lower than rejection for 25 nm particles.
25 𝑛𝑚 5
𝜆= = > 0.5
30 𝑛𝑚 6
so a correction equation must be applied….
2 2
𝑅𝑖 = 1 − (1 − (𝜆(2 − 𝜆)) ) 𝑒 −0.7146𝜆

20
 𝑅𝑖 = 0.9666470148
𝑂𝑣𝑒𝑟𝑎𝑙𝑙 𝑅𝑒𝑗𝑒𝑐𝑡𝑖𝑜𝑛 = (0.9666)(0.1429) + (1)(0.8571) = 0.9952352878
Disk Centrifuge Calculations
- Centrifuge specifications: Recover ~100% of the nanoparticle-encapsulated drug; rate-
limiting is smallest nanoparticles.
- Variable definitions:
o Q = volumetric flow rate
o a = radius of particle
o ρ = density of particle
o ρ0 = density of fluid
o μ = viscosity of fluid
o vg = sedimentation velocity
o g = gravity constant (9.81 m/s2)
o ω = rotational velocity
o n = number of stacks within disk centrifuge
o θ = angle of disk stacks with respect to center line of centrifuge
o Rl = radius from center line to inside of disk stacks
o R0 = radius from center line to outside of disk stacks
o Σ = operation constant of centrifuge

𝑄 = (𝑉𝑔 )(𝛴)

2𝑎2 (𝜌 − 𝜌0 )𝑔
𝑉𝑔 =
9𝜇
a = 12.5 nm = 12.5 x 10-9 m
ρ = 1.3018 g/cm3
ρ0 = 1 g/cm3
μ = 0.01 g/(cm*s)
2 𝑔 𝑔 𝑚 106 𝑐𝑚3
2(12.5×10−9 𝑚) (1.3018 −1 3 )(9.81 2)( )
𝑐𝑚3 𝑐𝑚 𝑠 𝑚3
𝑉𝑔 = 𝑔 = 1.02800625 x 10-8 cm/s
9(0.01 )
𝑐𝑚×𝑠

2𝑛𝜋𝜔2 (𝑅0 3 − 𝑅1 3 )(cot(𝜃))

𝛴=
3𝑔
𝑛 = 48 𝑠𝑡𝑎𝑐𝑘𝑠
2𝜋 𝑟𝑎𝑑𝑖𝑎𝑛𝑠
𝜔 = 6240 𝑟𝑝𝑚 × ( ) = 39207.07632 𝑟𝑎𝑑𝑖𝑎𝑛𝑠/𝑚𝑖𝑛
𝑟𝑒𝑣𝑜𝑙𝑢𝑡𝑖𝑜𝑛
R0= 30 cm
R1 = 7 cm

21
 Θ = 45°

𝑟𝑎𝑑 2
2(48) (39207.07632 𝑚𝑖𝑛) ((30 𝑐𝑚)2 − (7 𝑐𝑚)2 )(cot(45°))
𝛴= = 371294621.6 𝑐𝑚2
𝑚 100 𝑐𝑚 60 𝑠 2
3 (9.81 2 ) ( 𝑐 ) ( 𝑚𝑖𝑛 )
𝑠

3.816931916 𝑐𝑚3 60 𝑠 229.0159149 𝑐𝑚3

𝑄 = (𝑉𝑔 )(𝛴) = ( )=
𝑠 𝑚𝑖𝑛 𝑚𝑖𝑛
Reasoning Behind Project Decisions
Decision not to add upstream proposal for production of PLGA:
During the process of the design, the team evaluated potentially designing an upstream
plant to provide their own PLGA. This was in part inspired by a metabolic engineering lecture given by
Pedro
Decision not to recycle:
In the beginning of the design phase, the team considered attempting to recycle the
products in order to retrieve unencapsulated drug. This would reduce the price of consumables
significantly, since Paclitaxel is $50 per gram from Phyton Biotech. However, the team decided not to
recycle the drug because of the uncertainty surrounding the safety and efficacy of the recycled drug. This
uncertainty is due in large part to the lack of knowledge regarding the pharmacological properties of the
recycled drug compared to drug introduced into the main production stream only once (Doctor Matt
Dickson and Doctor Anthony Tuesca, personal communication, 3 April 2018). If safety and efficacy
could be verified, then it would certainly make sense to recycle the drug and increase profitability, but the
team decided it would be best to proceed under the assumption that the drug could not be recycled. The
team also looked into recycling for water and acetone streams to see if it would reduce the cost of
manufacturing by reducing the cost of consumables. However, during the design phase the team quickly
realized that the cost of purification of the water and acetone streams would significantly outweigh the
return of reduced cost of consumables. This is in large part because there are three major components that
must be considered in the purification process, removal of particulates (free drug and PLGA encapsulated
Paclitaxel unintentionally removed from main product stream), separation of the miscible liquids water
and acetone, and sterilization of the water and acetone streams before reuse. The guidelines the team
considered fell under United States Pharmacopeia (USP section 1231 (US Pharmacopeia, 2018; Pharmate
Systems, 2018). The first step during the design phase of recycling is the removal of the particulate
matter. The team considered the removal of the particulates at first using a granular activated carbon
(GAC) bed, since they commonly are used in water purification particularly in accordance with USP and
EPA standards (US Pharmacopeia, 2018; EPA, 2018). However, the team found that the removal using a
GAC bed was time consuming and required an extensive amount of time to extract the components. The
team tried various empty bed contact times between 5 and 30 minutes as suggested (University of Sevilla,
2018; EPA, 2018). However, the team quickly found that in order to add enough units in order to keep up
with scheduling was incredibly expensive, and increasing the size of the GAC beds was also extremely
costly. The team tried not using a GAC bed and instead using two reverse osmosis feed and bleed unit
operations to remove particles prior to distillation to purify the water since reverse osmosis was also a
common process falling underneath USP guidelines (US Pharmacopeia, 2018). However, even with these
changes the team was not able to design a profitable process with water purification for return. The

22
conclusion the team came to with regards towards the recycle stream was that the cost of purification in
order to recycle water is somewhat dependent on the scale of the plant that would be constructed, and that
if the product line was part of a larger plant that water purification would become more profitable and
vital. This reasoning is guided by the increasing trend of water purification for reuse in large scale
industrial settings (Gunderson, 2018).
Economic Assumptions
The economic evaluation report (Appendix ?), gives a total capital investment of $ US dollar. The
operating cost of this project is $ USD per year, while revenue is $1,191,439,000 USD per year. Given
this, it is expected that the return on this investment will be 21.08% and the payback time is 3.10 years.
After taxes, the internal rate of return is 21.48% and the net present value at a 7% interest rate after 15
years is $2,068,267,000. These calculations are based on producing the finished product at a rate of
entities per year, which each entity producing a revenue of $16,700. The economic evaluation report
provides an overall analysis of labor, materials, and other costs. The main components that will be
flowing through the pipe are water, acetone, PLGA, and Paclitaxel. Based on production of the
comparable market drug Abraxane, there is a need to produce about 336.70 kg of product per year (see
appendix)
The cash flow analysis report, appendix?, provides an analysis from year 1 to year 15 and details
capital investment, sales revenue, operating costs, and other parameters per year. Year 5 is when sales
revenue begins. According to this analysis, the internal rate of return after taxes is 13.67%. At an interest
rate of 7%, the net present value at year 15 is $798,505. Loan payments of $38,440,176 occur annually
between year 5 and year 13. Direct fixed capital costs between years 1 and 3, total
The cash flow analysis report, appendix?, provides an analysis from year 1 to year 15 and details
capital investment, sales revenue, operating costs, and other parameters per year. Year 5 is when sales
revenue begins. According to this analysis, the internal rate of return after taxes is 13.67%. At an interest
rate of 7%, the net present value at year 15 is $798,505. Loan payments of $38,440,176 occur annually
between year 5 and year 13. Direct fixed capital costs between years 1 and 3, total

23