Doxorubicin and Anti-PD-L1 Antibody Conjugated Supra Iron

Oxide Magnetic Nanoparticles for Colorectal Cancer chemotherapy

1. Colorectal Cancer
Cancer is a major public health problem worldwide and is the second leading cause of death in
the United States (Rebecca L. Siegel, 2020) Colorectal cancer was the most common cause of
cancer death in the United States in the late 1940s and early 1950s. (Rebecca L. Siegel, 2020).
Today, it is the third leading cause of cancer death in both men and women, in part because of
historical changes in risk factors, the introduction and dissemination of early detection tests, and
improvements in treatment. (Rebecca L. Siegel, 2020)An annual report on the status of CRC
(issued by the American Cancer Society) documented five-year survival rates ranging from 88.1%
(stage I) to only 12.6% (stage IV. (Cronin, et al., 2018)
2. Chemotherapy
The most effective therapeutic strategy for all stages of localized CRC is surgical resection.
However, surgery may not eliminate all cancerous cells and around 50 percent of advanced-stage
CRC patients experience tumor regrowth and recurrence following surgery. (Cho, et al., 2017).
Therefore, the prognosis of patients with advanced CRC after surgery still remains poor. (Cho, et
al., 2017), (Manchun, Dass, Cheewatanakornkool, & Sriamornsak, 2015). Adjuvant treatments
administered after surgery, such as chemotherapy, have reduced recurrence and increased survival.
(Evans, et al., 2016). Chemotherapeutics like doxorubicin (DOX), fluorouracil, cisplatin, and
mitomycin are usually used to kill residual CRC cells after surgery. (Manchun, Dass,
Cheewatanakornkool, & Sriamornsak, 2015). Doxorubicin (Dox) is one of the most effective
anticancer drugs against many types of cancer tumors including colorectal cancer.DOX is an
anthracycline that inhibits nucleic acid synthesis (Rezazadeh, Akbari, Amuaghae, & Emami, 2018)
and inhibits the proliferation CRC cell (smorkamol manchun, 2015) However, the intravenous
administration of most classical chemotherapeutic drugs still monotherapies are unsatisfactory due
to the ability of CRC cells to develop multidrug resistance (Correale, et al., 2005) and undesirable
biodistribution produce unwanted effect to normal cell. (Sin-Tzu Ning, 2016) The drugs are rapidly
removed from the blood, and only a small amount of administered drugs reached to tumor site.
(Sin-Tzu Ning, 2016)
3. Targeted chemotherapy
The combinatorial strategies, such as, chemotherapy with immunotherapy (Correale, et al., 2005)
radiation therapy (Correale, et al., 2005) targeted therapies (Zhao, et al., 2015) photothermal
therapy (PTT) (Nam, Son, Ochyl, Kuai, & Schwendeman, 2018) have been reported to
cooperatively suppress cancer progression and to minimize side effects. (Zheng, et al., 2013). To
enhance chemotherapeutic drug accumulation in tumors, Nanodelivery systems such as liposomes,
micelles, nanotubes, nanogold and magnetic nanoparticle have been developed. (Anders, et al.,
2013)Through the enhanced permeation and retention (EPR) effect, these nanoparticles are
considered potential nanocarriers. (Greish, 2010). To achieve active targeting, which is also called
ligand mediated targeting, the surfaces of nanoparticles are modified with affinity ligands to
increase their uptake by specific cells. Appropriate ligands are selected to target specific molecules
that are overexpressed on the cell plasma membrane. The purpose of active targeting systems is to
increase specific binding of nanoparticles to the cells and enhance the internalization of delivered
drugs (Byrne, Betancourt, & Brannon-Peppas, 2008). In oncology, the development of targeted
nanoparticles can increase the internalization of drugs in targeted tumor cells, minimize drug
leakage from target sites, and protect the drug from degradation and elimination.6 (Lammers,
Hennink, & Storm, 2008). To date, many nanoparticles and magnetic nanoparticle have been
designed with conjugated ligands, including antibodies, carrier proteins, and peptides (Mahon,
Salvati, Baldelli Bombelli, Lynch, & Dawson, 2012) Of the many small molecules tested,
antibodies have shown good targeting specificity and affinity, offering preferable therapeutic
efficiency with fewer side effects for future clinical application. (Tazi, Nafil, & Mahmal, 2012).

Formation of Anti-PD-L1 Antibody Conjugated Iron Oxide Magnetic Nanoparticles

o Formation of magnetic Nano particle

Initially, the IOMN nanoparticles were synthesized using a modified pyrolysis-based method in
organic solvent. These hydrophobic core IOMN nanoparticles were converted into the biocompatible
water soluble form with the use of amphiphilic polymer that interacted with the original hydrophobic
oleate coating resulting in a double layered structure with carboxyl groups on the surface.
One of the major advantages for IOMN with PEG (IOMN -LA-PEG) as a drug carrier is that the
degradation of Dox could be effectively reduced by conjugating Dox on the surface of IOMN -LA-
PEG. The degree of freedom of Dox IOMN -LA-PEG was limited to prevent its molecular distortion
and hence increase its half-life. In addition, IO-LP-PEG- could cause DNA crosslink more serious,
resulting in a lower DNA expression and a higher CRC cells. apoptosis. Finally, the accumulation of
IOMN -LA-PEG in cancer tumors could be enhanced by applying a magnetic field in the tumor to
increase the degree of DNA crosslink and to reduce the damage of normal organs. (citation)

o Formation of LA-PEG-PD-L1

PD-L1 is a type I transmembrane protein that was recently implicated in the etiology of CRC and
shown to be a biomarker of CRC. (Luan, et al., 2016) Furthermore, PD-L1overexpression is known
to be significantly associated with prognosis and overall survival in curatively resected CRC
patients, (Morihiro, et al., 2017) and anti-PD-L1 antibody ie used her for target delivery to CRC.

LA-PEG-PD-L1 was conjugated by reacting the ε-amino groups on the lysine residue of the
anti-PD-L1 antibody with the α-terminal end group LA-PEG-NHS (Zhou, Zhang, Zhang, Ma,
& Su, 2016) through amide linkage. (Fakhrossadat Emami, 2019)
o Formation of LA-PEG-DOX

o LA-PEG-DOX is conjugated by reaction the amine group of DOX with the α-terminal end group the
LA-PEG-NHS (Zhou, Zhang, Zhang, Ma, & Su, 2016) through (−CO-NHR) amide linkage.
(Fakhrossadat Emami, 2019)

o Formation of DOX-LA-PEG-PD-L1

PEGylated anti-PD-L1 antibody (LA-PEG-PD-L1) were covalently attached to the surfaces of

IOMNP by dithiol-FE covalent bonds. AuNP were further coated with short PEG-SH chains to
improve nanoparticles (NP) stability. (23) (IOMNP by dithiol-FE covalent bonds).

la-polyethylene glycol modification of MNP improve the circulation time of therapeutic agents or
delivery devices. The hydrophilic PEG can improve the biocompatibility of the delivery system
because most of the biological environment is hydrophilic, and biocompatibility appears to be
 correlated with the degree of hydrophilicity exhibiting on the surface. (Po-Chin Liang, 2016) (Li,
Takashima, Yuba, Harada, & Kono, 2014).

a. Therapeutic action
i. Cytotoxicity
ii. mechanism
b. pros and cons
i. prose
ii. cons
4. reference
5.
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Cho, J., Lee, J., Bang, H., Kim, S. T., Park, S. H., An, J. Y., . . . Kim, k. (2017). Programmed cell death-ligand
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immunotherapy of metastatic colorectal carcinoma with gemcitabine plus FOLFOX 4 followed by
subcutaneous granulocyte macrophage colony-stimulating factor and interleukin-2 induces
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Correale, P., Cusi, M. G., Tsang, K. Y., Del Vecchio, M. T., Marsili, S., Placa, M. L., . . . al, e. (2005). .
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Pharm. Sci, 181.

Sin-Tzu Ning, S.-Y. L. (2016). Targeting Colorectal Cancer Stem-Like Cells with Anti-CD133. ACS Appl.
Mater. Interfaces, 17793-17804.

smorkamol manchun, c. R. (2015). Enhanced anti-tumor effect of pH-responsive dextrin nanogels

delivering doxorubicin on colorectal cancer. Carbohydrate Polymers, 222-230.

Tazi, I., Nafil, H., & Mahmal, L. (2012). Monoclonal Antibodies in Hematological Malignancies: Past.
Present and Future. J. Cancer Res, 281-269.

Zhao, L., Zhu, J., Cheng, Y., Xiong, Z., Tang, Y., Guo, L., . . . Zhao, J. (2015). Chlorotoxin-conjugated
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tomography imaging and radiotherapy of gliomas. ACS Appl. Ma, 19798-19808.

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Anders, C. K., Adamo, B., Karginova, O., Deal, A. M., Rawal, S., Darr, D., . . . Zamboni, W. C. (2013).
Pharmacokinetics and Efficacy of PEGylated Liposomal Doxorubicin in an intracranial Model of
brest cancr. PLoS ONE, 61369.

Byrne, J. D., Betancourt, T., & Brannon-Peppas, L. (2008). Active Targeting Schemes for Nanoparticle
Systems in Cancer Therapeutics. Adv. Drug Delivery Rev. 2008, 60 (15), 1615−1626, 1615-1626.

Cho, J., Lee, J., Bang, H., Kim, S. T., Park, S. H., An, J. Y., . . . Kim, k. (2017). Programmed cell death-ligand
1 expression predicts survival in patients with gastric carcinoma with microsatellite instability.
Oncotarget.

Correale, P., Cusi, M. G., Del Vecchio, M. T., Tsang, K. Y., Marsili, S., Placa, M. L., . . . all, e. (2005). Chemo-
immunotherapy of metastatic colorectal carcinoma with gemcitabine plus FOLFOX 4 followed by
subcutaneous granulocyte macrophage colony-stimulating factor and interleukin-2 induces
strong immunologic and antitumor activity in metastatic colon cance. J. Clin. Oncol., 8950-8958.

Correale, P., Cusi, M. G., Tsang, K. Y., Del Vecchio, M. T., Marsili, S., Placa, M. L., . . . al, e. (2005). .
Chemo-immunotherapy of metastatic colorectal carcinoma with gemcitabine plus FOLFOX 4
followed by subcutaneo factor and interleukin-2strong immunologic and antitumer activity in
metatastatic colon cancer patients. J. Clin. Oncol, 8950-8958.

Cronin, K. A., Lake, A. J., Scott, S., Sherman, R. L., Noone, A., Howlader, N., . . . Ma, J. ( 2018). Annual
Report to the Nation on the Status of Cancer part I. National cancer statistics. Cancer.

Evans, J. P., Sutton, P. A., Winiarski, B. K., Fenwick, S. W., Vimalachandran, D., Tweedle, E. M., . . .
Kitteringham, N. R. (2016). From mice to men Murine models of colorectal cancer for use in
translational research. Crit. Rev. Oncol./Hematol, 98 , 94−105.

Greish, K. (2010). . Enhanced Permeability and Retention (EPR) Effect for Anticancer Nanomedicine Drug
Targeting. Methods Mol. Biol, 25−37.

Lammers, T., Hennink, W. E., & Storm, G. (2008). Tumour-Targeted Nanomedicines: Principles and
Practice. Br. J. Cancer, 392-397.

Mahon, E., Salvati, A., Baldelli Bombelli, F., Lynch, I., & Dawson, K. A. (2012). Designing the Nanoparticle-
Biomolecule Interface For “targeting and Therapeutic Delivery. J. Controlled Release, 164-174.

Manchun, S., Dass, C. R., Cheewatanakornkool, K., & Sriamornsak, P. (2015). Enhanced anti-tumor effect
of pH-responsive dextrinnanogels delivering doxorubicin on colorectal cancer. Carbohydr.
Polym., 126, 222−230.

Nam, J., Son, S., Ochyl, L. J., Kuai, R., & Schwendeman, A. M. (2018). . Chemo-photothermal therapy
combination elicits antitumor immunity against advanced metastatic cancer. Nature
communacation.

Po-Chin Liang, C.-F. C.-R.-Y.-Y.-L. (2016). Doxorubicin-modified magnetic nanoparticles as a drug delivery
system for magnetic resonance imaging-monitoring magnet-enhancing tumor chemotherapy.
International Journal of Nanomedicine.
Rebecca L. Siegel, K. D. (2020). Cancer Statistics, 2020. CA CANCER J CLIN, VOLUME 70. Retrieved from
https://acsjournals.onlinelibrary.wiley.com/doi/full/10.3322/caac.21590

Rezazadeh, M., Akbari, V., Amuaghae, E., & Emami, J. (2018). Preparation and characterization of an
injectable thermosensitive hydrogel for simultaneous delivery of paclitaxel and doxorubicin. Res.
Pharm. Sci, 181.

Sin-Tzu Ning, S.-Y. L. (2016). Targeting Colorectal Cancer Stem-Like Cells with Anti-CD133. ACS Appl.
Mater. Interfaces, 17793-17804.

smorkamol manchun, c. R. (2015). Enhanced anti-tumor effect of pH-responsive dextrin nanogels

delivering doxorubicin on colorectal cancer. Carbohydrate Polymers, 222-230.

Tazi, I., Nafil, H., & Mahmal, L. (2012). Monoclonal Antibodies in Hematological Malignancies: Past.
Present and Future. J. Cancer Res, 281-269.

Zhao, L., Zhu, J., Cheng, Y., Xiong, Z., Tang, Y., Guo, L., . . . Zhao, J. (2015). Chlorotoxin-conjugated
multifunctional dendrimers labeled with radionuclide 131I for single photon emission computed
tomography imaging and radiotherapy of gliomas. ACS Appl. Ma, 19798-19808.

Zheng, M., Yue, C., Ma, Y., Gong, P., Zhao, P., Zheng, C., . . . Cai, L. (2013). . Single-step assembly of DOX/
ICG loaded lipid−polymer nanoparticles for highly effective chemophotothermal combination
therapy. ACS Nano 2013, 2056.

Refrence
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thermosensitive hydrogel for simultaneous delivery of paclitaxel and doxorubicin. Res. Pharm. Sci. 2018, 13 (3),
181.
(6) Manchun, S.; Dass, C. R.; Cheewatanakornkool, K.; Sriamornsak, P. Enhanced anti-tumor effect of pH-responsive
dextrin nanogels delivering doxorubicin on colorectal cancer. Carbohydr. Polym. 2015, 126, 222−230 (smorkamol
manchun, 2015)
(12) Correale, P.; Cusi, M. G.; Tsang, K. Y.; Del Vecchio, M. T.; Marsili, S.; Placa, M. L.; Intrivici, C.; Aquino,
A.; Micheli, L.; Nencini,C.; et al. Chemo-immunotherapy of metastatic colorectal carcinoma with gemcitabine plus
FOLFOX 4 followed by subcutaneous granulocyte macrophage colony-stimulating factor and interleukin-2 induces strong
immunologic and antitumor activity in metastatic colon cancer patients. J. Clin. Oncol. 2005, 23 (35), 8950−8958. (Correale,
et al., 2005)
(17) (17 (Nam, Son, Ochyl, Kuai, & Schwendeman, 2018).
12…….Zhao, L.; Zhu, J.; Cheng, Y.; Xiong, Z.; Tang, Y.; Guo, L.; Shi, X.; Zhao, J. Chlorotoxin-conjugated multifunctional
dendrimers labeled with radionuclide 131I for single photon emission computed tomography imaging and radiotherapy of
gliomas. ACS Appl. Mater. Interfaces 2015, 7 (35), 19798−19808 (Zhao, et al., 2015)
18… (18) Zheng, M.; Yue, C.; Ma, Y.; Gong, P.; Zhao, P.; Zheng, C.; Sheng, Z.; Zhang, P.; Wang, Z.; Cai, L. Single-step
assembly of DOX/ ICG loaded lipid−polymer nanoparticles for highly effective chemophotothermal combination therapy. ACS
Nano 2013, 7 (3), 2056− 2067. (Zheng, et al., 2013)

(1) Anders, C. K.; Adamo, B.; Karginova, O.; Deal, A. M.; Rawal, S.; Darr, D.; Schorzman, A.; Santos, C.; Bash, R.; Kafri, T.;
Carey, L.; Miller, C. R.; Perou, C. M.; Sharpless, N.; Zamboni, W. C. Pharmacokinetics and Efficacy of PEGylated Liposomal
Doxorubicin in an Intracranial Model of Breast Cancer. PLoS One 2013, 8 (5), e61359. (Anders, et al., 2013)
(4 (Greish, 2010)