RESEARCH

PROPOSAL
The Clinical possibilities of synthetic zeolites and mesoporous silica
as controlled-release drug delivery systems for anti-cancer drugs.

Sami Miah
UP764563

Word count: 2,197 (not including headings, sub-headings and references).

Supervisor: Dr. Paul Cox

The Clinical possibilities of synthetic zeolites and mesoporous silica as
controlled-release drug delivery systems for anti-cancer drugs.

Table of Contents
1. BACKGROUND 3

1.1: INTRODUCTION……………………………………………………………………… 3
1.2: 5-FLUOROURACIL (5-FU) AND ITS LIMITATION……………………………………… 3
1.3: ZEOLITES AND MESOPOROUS SILICA: FUTURE DRUG-DELIVERY SYSTEMS…………… 5
1.4: EFFECTS OF CONTROLLED DRUG RELEASE WITH DIFFERENT ZEOLITE STRUCTURAL
FRAMEWORK AND CHANNEL SIZE……………………………………………………. 7

2. AIMS AND OBJECTIVES 8

2.1: AIMS………………………………………………………………………………… 8
2.2: OBJECTIVES…………………………………………………………………………. 8

3. METHODOLOGY 9

3.1: MATERIALS…………………………………………………………………………... 9
3.2: ENCAPSULATION OF 5-FU WITHIN FRAMEWORK (DRUG LOADING VIA SOAKING
PROCEDURE) AND DETERMINATION OF 5-FU LOADED WITHIN FRAMEWORKS………… 9
3.3: CHARACTERISATION OF MICROPOROUS AND MESOPOROUS FRAMEWORK-TYPES……... 10
3.4: DETERMINATION OF 5-FU RELEASE FROM FRAMEWORKS VIA DISSOLUTION STUDIES… 10
3.5: STATISTICAL ANALYSIS………………………………………………………………. 11
3.6: SCREENING FOR OTHER POSSIBLE FRAMEWORK SYSTEMS…………………………….. 12
3.7: ETHICAL CONSIDERATIONS…………………………………………………………... 12
3.8: POTENTIAL FUTURE WORK……………………………………………………………. 12

4. ESTIMATED COSTS 13

5. ANTICIPATED OUTCOMES 14

6. REFERENCES 16

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1. Background
1.1: Introduction.

One of the most prominent chronic conditions on the rise is cancer. Cancer is the

abnormal cell growth, which has the potential to spread throughout the body1. Newly

diagnosed cases of cancer in England continued to rise in 2015 with almost 300,000

cancers registered2. Colorectal cancer (CRC) came up as the third most common

cancers with 34,729 cases being accounted for in 20152.

CRC a type of cancer that begins either in the colon or the rectum. The development is

slow, taking a period of 10-20 years, but early detection makes CRC curable3. Current

treatments include surgery, radiotherapy, chemotherapy and targeted therapy. One of

the drugs used in CRC Stage-II chemotherapy regiment is the cytotoxic drug

Fluorouracil (5-FU)4. Cytotoxic drugs treat cancer by being toxic to cells that are

proliferating at a faster rate than normal cells (i.e. cancer cells, since they undergo rapid

cellular growth and division). 5-FU is a pyrimidine analog that is an antineoplastic

antimetabolite (stops the cells making and repairing DNA). Ultimately, DNA synthesis

is interfered by the inhibition of thymidylate synthetase conversion of deoxyuridylic

acid to thymidylic acid5.

1.2: 5-Fluorouracil (5-FU) and its limitations.

A key issue with cytotoxic anti-cancer drugs is their lack of specificity of their systemic

bio-distribution and the number of side-effects that patients experience due to the drug

attacking both normal and cancerous cells6. They also exhibit unfavorable

pharmacokinetic characteristics and thus resulting in higher doses being administered to

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achieve correct therapeutic levels. Additionally, they are also prone to rapid clearance

and their in-vivo degradation results in their necessity of high dose administration6.

The major limitation of 5-FU is its high rate of metabolism in the body and as a result to

maintain the correct therapeutic serum concentration level, it requires numerous high

dose administration6. In addition, since 5-FU has a very narrow therapeutic window,

serum concentration above maximum tolerated dose can result in severe toxic effects7.

5-FU also has a considerably low half-life of only 10-20 minutes, as well as rapid

clearance where over 90% is excreted in the first hour5. Both these characteristics result

in patients having to be administered more frequent doses. 5-FU also has an 8-12% low

protein binding figure5. As a result, due to its low protein-binding characteristic, more

free drugs (with potential for inducing its biological activity) are able to efficiently

traverse into cell membranes or diffuse into them6. When it comes to cytotoxic drugs

this can be a problem as you come into issues such as causing a negative effect on

normal non-cancerous cells. There are potential possibilities to improve the therapeutic

efficiency of 5-FU and overcome the therapeutic toxicity issue that results in a number

of side-effects. Also with possibilities to enhance and prolong the specific accumulation

of cytotoxic agents in tumor/cancerous cells specifically6. A potential delivery system

for 5-FU should possess the following characteristics; it must be stable, the size should

be small to allow capillary distribution of the drug and a uniform perfusion at the

desired target site (i.e. malignant cancerous cells); should be able to load an adequate

amount of 5-FU; a form of protection against degradation and most importantly be able

to release 5-FU at a controllable rate of release to the desired target site.

Controlled-release is integral in the future of effective chemotherapy treatment since

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one of the major obstacles in every regiment is to always provide the proper therapeutic

dose of the cytotoxic agent at only the desired locals of action, while minimising the

side-effects. This can be difficult, but one drug delivery system that shows huge

potential are microporous and mesoporous materials. The use of these in clinical

settings could have the potential to reduce the need for multiple does, by instead giving

a single dose that releases the drug at a controlled-rate of release. This would eliminate

both the cost and the management of therapeutic dosing, thus resulting in huge cost

savings for the NHS.

1.3: Zeolites & Mesoporous silica: Future drug-delivery systems.

Zeolites are microporous structures that posses channel-like pore systems of diameters

< 2nm, while mesoporous materials like mesoporous silica have pore diameters of

between 2nm and 50nm8. Both materials exhibit a uniform network of 3-dimensional

channel systems. They are regular crystal channel structures that are reproducible.

Zeolites are inorganic crystalline aluminosilicate structures consisting of the elements;

Si, Al and O as the fundamental atoms making up their structure (Figure 1)8.

They exhibit a tetrahedra structure that is built of [AlO4]5 and [SiO4]48. This results in

the formation of structures that contain long cavities/channels of various sizes, which

have the capacity to allow small molecules to easily drift into and out of the framework.

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The presence of AlO4 in the framework results in a negative charge, thus either group 1

(Na+) or group 2 (Ca2+) must be in incorporated within the structure to ensure a neutral

charge and overall stability of the crystal.

Three synthetic zeolites that are of importance are; FAU, MFI and EUO. FAU

(faujasite) one of many frameworks of zeolites consists of sodalite cages that are

connected via hexagonal prisms. It is in these cages that drugs can be loaded. This

results in the formation of a reproducible channel system that is unique to this FAU type

zeolite. FAU’s per unit cell exhibit 208 channels and 32 cages9. Its pore and channel

system characteristics are shown in Table 1.

MFI, another zeolite, has 81 channels and 12 cages10, while EUO contains 96 channels

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and 20 cages per unit cell11. Both these are small structures compared to FAU resulting

in smaller pore diameters (see Table 2 & 3). But this could be a potential factor at a

slower rate of release.

Mesoporous frameworks such as MCM-41 and SBA-15 are also potential system but

these possess larger channels and even though they may have the capacity for higher

drug loading, they may be limited in terms of controlled release due to their large

pores/channels21. But both frameworks have the potential for external surface

modification in order to modify its porosity due to the presence of silanol groups20,21.

1.4: Effects of controlled drug release with different zeolite structural

framework and channel size.

Due to these structural characteristics zeolites posses,

they can be used to load drugs into their framework

(Figure 2). Depending on the shape and size of the

channels the controlled-rate of release can be adjusted. If

the channels are too big for a small drug like 5-FU, then

there will be very limited interactions between the

channel walls resulting in a fast rate of release and vice

versa. Thus the degree of interaction between 5-FU and

channel size and its properties shows great possibilities

in terms of controlling the rate of release of the drug with

possible hopes in reducing clinical side-effects

associated with 5-FU therapy.

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2. Aims and Objectives

2.1: Aims.

The aims of this study are to identify a suitable zeolite/mesoporous-silica framework-

type that will successfully deliver 5-FU at a sustained control-rate to improve

chemotherapeutic treatment of CRC.

2.2: Objectives.

1. To identify a potential zeolite (FAU and MFI) or mesoporous-silica (MCM-41 and

SBA-15) for 5-FU sustained controlled release and if their channel/pore sizes and

framework configuration affect controlled-release.

2. Enhance the efficacy of 5-FU in terms of improving the bio-distribution and

pharmacokinetic characteristics (i.e. clearance) by identifying the most suitable

zeolite for encapsulation.

3. Improve the clinical management by reducing the frequent administration of 5-FU

by exploiting the characteristics of zeolites to achieve a sustained controlled-rate of

release.

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3. Methodology

3.1: Material.

Two zeolite framework-types (FAU and MFI) and two mesoporous-silica framework-

types (MCM-41 and SBA-15) will be used to identify the best candidate for the

sustained controlled release of 5-FU. These will all be obtained from commercially

available supplier Merch Sigma-Aldrich®, and so will 5-Fluorouracil and all other

chemicals and solvents as needed.

3.2: Encapsulation of 5-FU within framework (drug loading via soaking

procedure) and determination of 5-FU loaded within frameworks.
Method adapted from reference15.

Loading of 5-FU into frameworks will be achieved as follows: 20g of each framework-

type will be soaked (minimum 2 days, at room temperature, under continuous stirring)

in a 5g solution of 5-FU in methanol (300ml). Lastly, the removal of the solvent via

filtration using a 0.22µM polyester filter paper and the samples of each encapsulated

framework will be dried by rotoevaporation under vacuum, at 30°C15. Loading of 5-FU

within the frameworks will be determined via thermogravimetry analysis (TGA). This

will be conducted at a heating rate of 10oC/min under a nitrogen purge of 40ml/min,

from 30 to 600oC15. This heats each of the encapsulated 5-FU framework over a period

of time and as temperature changes the outcome will be a determination of how much 5-

FU has been loaded into each framework as well as other characteristics such as; phase

transitions, absorption, desorption and thermal decomposition16.

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3.3: Characterisation of microporous and mesoporous framework-types.

Further assessment of the loading efficiencies of each framework samples will be

carried out using fourier-transform infrared spectroscopy (FTIR). A KBr plate will be

prepared using 10mg of each framework sample and 100mg of KBr and then will be

ground (2-5min)15. Spectroscopy pellets will be made using pellet-press dies and placed

in a spectrometer to obtain an IR spectra15.

Using SEM (scanning electron microscopy) morphological studies will be determined

to obtain surface topography and composition of each framework. Framework samples

will be gold-plated before the imaging process15.

XRD (X-ray diffraction) patterns will be obtained of each framework using a Siemens-

D5000 that consists of a PW3710 diffractometer and an X-ray tube (30mA and 40kV)

with a copper anode. This will be done at 0.033 degrees per second scanning rate at

diffraction angle 20, between a 5–35 degree range15.

Brunauer–Emmett–Teller (BET) analysis will also be conducted to determine surface

area (SA), pore size distribution as well as to predict the dissolution rate. This is useful

as dissolution rate is proportional to the specific SA and therefore the SA can be

exploited in predicting the bioavailability of 5-FU18. This will be done via

Quantachrome Autosorb 1-MP using N2 adsorption15.

3.4: Determination of 5-FU release from frameworks via dissolution studies.

The release of 5-FU from each framework will be determined by in vitro dissolution

basket method (100 rpm and 37 ± 0.5oC). The dissolution environment will contain

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900mL HCl 0.1M (pH 1.2) to simulate gastric juice and phosphate buffer 0.2M (pH 7.4)

to simulate intestine fluid. These conditions will respectively mimic mouth to colon

transit conditions of the GI tract region. Weighed amounts of loaded framework

samples (∼ 500mg) of 5-FU will be tableted (via compression) and placed in basket. At

each interval (15, 30, 45 and 60min), samples will be collected by a peristaltic pump

and released 5-FU concentration determined using spectrophotometer at λ max

266nm15,19.

3.5: Statistical analysis.

Both dissolution profiles will be compared using a simple model-independent

approach defined the difference factor (f1) and similarity factor (f2)15. This statistical

analysis is recommended for similarity assessment of two in vitro dissolution

profiles. The following are used (Equation 1 & 2)15.

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3.6: Screening for other possible framework systems.

Other frameworks can be determined as possible candidates for 5-FU control-release

using both molecular modelling and dynamics (i.e. VMD®)17. The later can be used to

simulate to see how fast 5-FU could potentially move across channel systems of many

framework-types.

3.7: Ethical Considerations.

Due to exclusion of the use of animal within this study, it will not require authorisation

from the University of Portsmouth’s Animal Research Committees and thus no ethical

review is deemed necessary.

3.8: Potential future work.

The magnetic properties of the frameworks could be introduced by potentially

incorporating atoms such as iron that display ferromagnetism properties. These

characteristics could be exploited with the use of an external device that applies a

magnetic felid which could ultimately drive the magnetic 5-FU encapsulated framework

to the desired location and thus further reducing the therapeutic limitation of 5-FU

chemotherapy6.

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4. Estimated costs

Estimation cost of chemicals and materials required are illustrated in Table 4.

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5. Anticipated outcomes

Analysis of each of the microporous and mesoporous framework-types will allow for

the successful identification of a controlled release system for 5-FU. Drug loading

capacity, via TGA studies, will identify and confirm the total amount of drug loaded

within each framework. FAU is anticipated to accommodate 5-FU more efficiently due

to increased range of sodalite cage sizes, greater abundance in pores (of larger diameter)

and channel systems.

FTIR, SEM, XRD and BET analysis will be performed to further confirm drug

encapsulation within each framework. BET Nitrogen absorption will reveal insights in

regard to surface-area (SA). Reduction in SA (post drug loading) will confirm

encapsulation, particular within the pores. XRD patterns of each framework (pre and

post drug loading) is expected to be similar as framework structure is not anticipated to

be affected via encapsulation of 5-FU. However certain peeks are expected to display

different intensities after drug loading, due to reduction in crystallinity after

encapsulation. Spectroscopic analysis, via FTIR, will determine the presence of 5-FU

encapsulation and interaction within each framework. Presence of 5-FU will reveal

bands at 3053cm-1, 3014cm-1 and 2825cm-1 due to both aromatic and aliphatic C-H

stretching vibrations. C-N stretching vibrations will be seen at band 1236cm-1 and C-F

stretching vibrations will be observed at 806cm-1. Confirmation of the integrity of

encapsulated 5-FU thus will be confirmed. Basket dissolution studies will provide drug

delivery profiles at two pH environments, with an expectation of increased dissociation

of 5-FU at higher pH, thus higher concentration detected.

An anticipated hypothesis is candidates that exhibit morphological features like large

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numbers of cages, channels and pore size (i.e. FAU type) to both display a larger

facilitation of 5-FU encapsulation while also maintaining sustained controlled release at

higher pH environments. This is important for future 5-FU therapy due to the

limitations that it faces clinically.

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6. References

1. Cancer - NHS Choices [Internet]. Nhs.uk. 2017 [cited 28 October 2017].

Available from: https://www.nhs.uk/Conditions/Cancer/Pages/Introduction.aspx

2. Cancer Registration Statistics, England - Office for National Statistics [Internet].

Ons.gov.uk. 2017 [cited 28 October 2017]. Available from:

https://www.ons.gov.uk/peoplepopulationandcommunity/healthandsocialcare/co

nditionsanddiseases/bulletins/cancerregistrationstatisticsengland/2015

3. Chandran S, Natarajan S, Chandraseharan S, Mohd Shahimi M. Nano drug

delivery strategy of 5-fluorouracil for the treatment of colorectal cancer. Journal

of Cancer Research and Practice. 2017;4(2):45-48.

4. Colorectal cancer - NICE Pathways [Internet]. Pathways.nice.org.uk. 2017 [cited

28 October 2017]. Available from:

https://pathways.nice.org.uk/pathways/colorectal-

cancer#path=view%3A/pathways/colorectal-cancer/managing-local-colorectal-

tumours.xml&content=view-node%3Anodes-further-treatment

5. Fluorouracil - DrugBank [Internet]. Drugbank.ca. 2017 [cited 29 October 2017].

Available from: https://www.drugbank.ca/drugs/DB00544

Sami Miah | UP764563 16  

6. Arias J. Novel Strategies to Improve the Anticancer Action of 5-Fluorouracil by

Using Drug Delivery Systems. Molecules. 2008;13(10):2340-2369.

7. Gamelin E, Boisdron-Celle M. Dose monitoring of 5-fluorouracil in patients with

colorectal or head and neck cancer—status of the art. Critical Reviews in

Oncology/Hematology. 1999;30(1):71-79.

8. ASDN - Chemistry - Zeolites [Internet]. Asdn.net. 2017 [cited 7 November

2017]. Available from: http://asdn.net/asdn/chemistry/zeolites.php#ref

9. ZEOMICS - Zeolite Pore Characterization of FAU (Faujasite) [Internet].

Helios.princeton.edu. 2017 [cited 10 November 2017]. Available from:

http://helios.princeton.edu/zeomics/cgi-bin/view_structure.pl?src=iza&id=FAU

10. ZEOMICS - Zeolite Pore Characterization of MFI (ZSM-5) [Internet].

Helios.princeton.edu. 2017 [cited 10 November 2017]. Available from:

http://helios.princeton.edu/zeomics/cgi-bin/view_structure.pl?src=iza&id=MFI

11. ZEOMICS - Zeolite Pore Characterization of EUO (EU-1) [Internet].

Helios.princeton.edu. 2017 [cited 10 November 2017]. Available from:

http://helios.princeton.edu/zeomics/cgi-bin/view_structure.pl?src=iza&id=EUO

Sami Miah | UP764563 17  

12. Faujasite Type Zeolite BCR704 [Internet]. Sigma-Aldrich. 2017 [cited 15

November 2017]. Available from:

https://www.sigmaaldrich.com/catalog/product/sial/bcr704?lang=en&region=G

13. Silica, mesostructured 643645 [Internet]. Sigma-Aldrich. 2017 [cited 15

November 2017]. Available from:

http://www.sigmaaldrich.com/catalog/product/aldrich/643645?lang=en&region=

GB&gclid=EAIaIQobChMIx6ioqYrB1wIV1ecbCh1mYw8ZEAAYASAAEgJM

t_D_BwE

14. Silica 806803 [Internet]. Sigma-Aldrich. 2017 [cited 15 November 2017].

Available from:

https://www.sigmaaldrich.com/catalog/product/aldrich/806803?lang=en&region

=GB

15. Khodaverdi E, Honarmandi R, Alibolandi M, Rafatpanah Baygi R, Hadizadeh F,

Zohuri G. Evaluation of synthetic zeolites as oral delivery vehicle for anti-

inflammatory drugs [Internet]. PubMed Central (PMC). 2017 [cited 15

November 2017]. Available from:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4069848/

16. Coats A, Redfern J. Thermogravimetric analysis. A review. The Analyst.

1963;88(1053):906.

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17. Humphrey W, Dalke A, Schulten K. VMD: Visual molecular dynamics. Journal

of Molecular Graphics. 1996;14(1):33-38.

18. BET – Particle Analytical [Internet]. Particle.dk. 2017 [cited 18 November

2017]. Available from: http://particle.dk/methods-analytical-laboratory/surface-

area-bet-2/

19. Chickpetty S, Raga B. Formulation, in vitro drug release and in vivo human X-

ray investigation of polysaccharide based drug delivery systems for targeting 5-

fluorouracil to the colon. Brazilian Journal of Pharmaceutical Sciences.

2013;49(2):263-273.

20. Ramírez A, Lopez B, Sierra L. Study of the Acidic Sites and Their

Modifications in Mesoporous Silica Synthesized in Acidic Medium under

Quiescent Conditions. The Journal of Physical Chemistry B.

2003;107(35):9275-9280.

21. Datt A. Applications of mesoporous silica and zeolites for drug delivery

[Internet]. Ir.uiowa.edu. 2012 [cited 23 November 2017]. Available from:

http://ir.uiowa.edu/cgi/viewcontent.cgi?article=3443&context=etd

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