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Biodistribution of monoclonal antibody nimotuzumab labeled 131I on nude mice bearing human head and neck cancer

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Research target: We conducted the research to determine the distribution of 131I-nimotuzumab through the indication of the radioactivity in tissues of nude mice bearing human head and neck cancer tumors.

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BIODISTRIBUTION OF MONOCLONAL ANTIBODY NIMOTUZUMAB

AND NECK CANCER

Nguyen Thi Kim Huong*; Ho Anh Son**; Nguyen Thi Thu*** Pham Huy Quyen*; Nguyen Linh Toan**

SUMMARY

Research target: We conducted the research to determine the distribution of 131 I-nimotuzumab through the indication of the radioactivity in tissues of nude mice bearing human head and neck

cancer tumors Subject and methods: The 131 I-nimotuzumab was injected into the mice’s veins

at the dose of 100 µCi per mouse, and radioactivity was counted at three time points: 24, 48, and 72 hours after the injection Results: The research results showed that all the tissues and organs manifested the radioactivity In tumor, the radioactivity count was higher than that of any other organs The highest indication was 48 hours after the injection, and then gradually declined Radioactivity of the tumor was 79,26% compared to blood count at 72 hours after the injection Conclusion: With blood normalize, distribution of 131 I-nimotuzumab in tumors is the highest compared with other tissues and organs of the nude mice

* Key words: Head and neck cancer; Monoclonal antibody nimotuzumab labeled 131 I; Nude mice

INTRODUCTION

Head and neck cancer is a disease

with high incidence in the world, as well

as in Vietnam, about 10% of annual cancer

patients diagnosed with head and neck

cancer among total number of cancer

patients [1] Head and neck cancer is

varied and complex, expressed in multiple

locations, offices, including throat cancer,

mouth floor, lowering the larynx and throat,

salivary glands, tongue, thyroid , causing

the rear significant effects on health, the

patient lives if not diagnosed early and

treated promptly Head and neck cancer

has higher expression amount of EGFR

(EGFR: Epidermal Growth Factor Receptor)

in an unusual way, is one of the important pathogenetic factors of growth, invasion excessive encroachment cancer cells in multiple parts of the body [4] Previous studies have confirmed that blocking EGFR will inactivate or prevent the growth and metastasis of cancer cells, allowing the treatment to be effective One of the products targeted for treatment purposes

is 131I-nimotuzumab To evaluation effect

of 131I-nimotuzumab on head and neck cancer, in this study, we measured the distribution of 131I-nimotuzumab in the tissues of nude mice bearing human head

and neck cancer

* Institute of Nuclear Research

** Vietnam Military Medical University

*** Haiphong Pharmaco-Medical University

Corresponding author: Ho Anh Son (hoanhsonhp@gmail.com)

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SUBJECTS AND METHODS

1 Materials, chemicals

- Nimotuzumab monoclonal antibodies

labeled with 131I radioisotope, produced at

the Institute of Nuclear Research, radioactive

concentrations 100 mCi/mL

- Gamma counting camera

2 Research subjects

BALB/c immune deficiency mice without

T lymphocytes (nude mice, Foxn1nu)

imported from Charlie - River Company

(USA) They were kept in clean rooms,

and filtered air with positive pressure

Room temperature is maintained at 25 ±

20C and humidity of 55 ± 5%, the light is

automatically switched control at 7h00, off

at 19h00 Food (Zeigler, USA) and sterilized

water were ad libitum Each mouse cage

is put on a track system with independent

ventilation and membrane filtration ensures

the good isolation with pathogens

Mice transplanted larynx cancer cells

Hep 2 (106 cells/mouse), after tumor reached

diameter of about 10 mm, then each animal

was injected 100 µL 131I-nimotuzumab

(100 µCi each) and randomly divided into

3 groups:

+ Group 1 (n = 6): Radiation measurements

24 hours after injection

+ Group 2 (n = 6): Radioactivity measured

48 hours after injection

+ Group 3 (n = 6): Radioactivity measured

72 hours after injection

3 Research method

- Mice took lugol 1%, 1 drop/head, 24 hours

before injecting 131I-nimotuzumab complex

to block the thyroid gland, and then, daily ingested water mix with lugol (1 drop of lugol/5 mL water)

- 131I-nimotuzumab (100 Ci) was injected into mouse tail vein [2] Then, animals were deeply anesthetized and internal organs like liver, spleen, kidneys, heart, blood, muscle, intestine, lung, tumor and thyroid gland were removed to weigh and measure radiation in order to calculate the distribution for each tissue

RESULTS

1 Distribution of 131 I-nimotuzumab

in tissues of the mice

in tissues of mice after 24, 48 and 72 hours (counts/mg)

n = 18 ORGANS

AVERAGE RADIATION COUNTS

(10s)

Results of biological distribution in mice showed that 131I-nimotuzumab radioactive compound distributed the highest in the blood at all the times and tend to decrease

at the time 48 and 72 hours

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200.00

400.00

600.00

800.00

1000.00

1200.00

Blood Tumor Muscle Heart Lung Kidney Liver Bowel Thyroid Spleen

ave24g ave48g ave72g Activation of beta rays in tissues (count x 10 3 /mg/10sec)

The count in tumor tissue was the highest at 48 hours and equivalent compared with lung Other organ counts were lower than these two Radiation activity in other tissue was lower than those in tumor and lung

2 Comparing 131 I-nimotuzumab distribution in tissues compared to blood

Table 2: Radiation counts of tissues compared with blood

Biodistribution of 131I-nimotuzumab was the highest in blood at all time points and gradually reduced by the time

Ave 24g Ave 48g Ave 72g

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20%

40%

60%

80%

100%

72g 48g 24g Counting ratios of tissues compared with the blood

Figure 2: Percentage of organs compared with the blood of nude mice

At the time 48 and 72 hours, normalize by blood counts, the radial activity of tumor

was the highest compared to other tissues

DISCUSSION

The distribution of radioactivity on the

tissues, including human head and neck

cancer tumors in nude mice showed that

it was expressed in all examined tissues,

and the ratios were different among

tissues, especially highest in the blood

(1114.42 μCi/g) and lowest in muscle

(95.94 μCi/g) at 24 hours after injection,

and then the radioactivity tends to decrease

at 48 hours and 72 hours after injection

After injection of 131I-nimotuzumab, antibody

molecules were concentrated in the blood

and had the highest ratio, then they will

move to the target tissue and attached to

its EGFR, so radioactivity decreased due

to degredation and elimination

On the contrary, in tumor, radioactive

count of 131I-nimotuzumab increased

gradually in 48 hours and it was higher

than other tissues which expressed high levels of EGFR in tumors, as other studies had demonstrated [4, 5], is the destination for the 131I-nimotuzumab attaches to and focus there, making the radiation increase

in tumor

High degree of EGFR expression in cancer cells is believed to be associated with increased levels of malignant progression as fast, easy-invasive and prone to relapse, but at the same time it is

a favorable factor for therapy destination and treatment are much better prospects

of therapeutic monoclonal antibodies used

in combination with radiation and anti-cancer

chemicals [3]

CONCLUSION

131I-nimotuzumab is showed in tissues after intravenous injection 24 hours and counting radioactive ratios in tissues were

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not equal, the highest expression levels in

time 48 hours after injection and then

decreased gradually Distribution of

131I-nimotuzumab in tumors compared with

blood was the highest in compare to other

tissues at 48 hours At 72 hours, it reached

79.26% against in blood

REFERENCES

1 University of Medicine and Pharmacy in

Hochiminh City Thematic oncology, Cancer

Prevention Workshop Hochiminh City 13th

2010, Appendix, Vol 14, No 4

2 Gopal B Saha Fundamentals of Nuclear

Pharmacy Sixth Edition Springer 2012

3 Denis Rolando Beckford Vera, Sebastian Eigner, Milos Beran, Katherina Eigner Henke, Alice Laznickova, Milan Laznicez, Frantisek Melichar, and Marco Chinol Preclinical evaluation

of 177Lu-nimotuzumab: a potential tool for radioimmunotherapy of epidermal growth factor receptor over expressing tumors cancer Biotherapy and Radiopharmaceuticals 2011, Vol 26, No 3

4 Herbst RS Review of epidermal

growth factor receptor biology Int J Radiat Oncol Biol Phys 2004, 59

5 Ariel Talavera, Rosmarie Friemann, Silvia Gómez-Puerta et al Nimotuzumab, an antitumor

antibody that targets the epidermal growth factor receptor, blocks ligand binding while permitting the active receptor conformation Cancer Res 2009

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