Small antibody fusion proteins with complementarity‑determining regions and lidamycin for tumor targeting therapy

Zhong,Gen-Shen; Wu,Min-Na; Guo,Xiao-Fang; Xu,Zhi‑Shan; Zhang,Sheng-Hua; Zhen,Yong-Su

doi:10.3892/ol.2013.1143

April 2013 Volume 5 Issue 4

Full Size Image

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

An International Open Access Journal Devoted to General Medicine.

April 2013 Volume 5 Issue 4

Full Size Image

Article

Small antibody fusion proteins with complementarity‑determining regions and lidamycin for tumor targeting therapy

Authors:
- Gen-Shen Zhong
- Min-Na Wu
- Xiao-Fang Guo
- Zhi‑Shan Xu
- Sheng-Hua Zhang
- Yong-Su Zhen
View Affiliations / Copyright

Affiliations: The First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan 453100, P.R. China, Xinxiang Medical University, Xinxiang, Henan 453003, P.R. China, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, P.R. China
Pages: 1183-1188
|
Published online on: January 18, 2013

https://doi.org/10.3892/ol.2013.1143
Expand metrics +

Abstract

Gelatinases are overexpressed in several types of maligancies and tumor stromal cells. Lidamycin is an enediyne antitumor antibiotic, which is composed of an apoprotein (LDP) and an active chromophore (AE). It is known that the heavy-chain complementarity-determining region-3 (CDR3) domain of scFv is important in antibody affinity. The aim of this study was to prepare the enediyne‑energized fusion proteins with a heavy-chain CDR3 domain of anti-gelatinases scFv and lidamycin, and to evaluate their antitumor efficiency. Fusion proteins comprising the CDR3 domain and the lidamycin apoprotein were generated, and ELISA, immunofluorescence and FACS were used to analyze the binding of the fusion protein with antigen gelatinases. The purified fusion proteins were assembled with the lidamycin chromophore, and the antitumor effects were evaluated in vitro and in vivo. It was found that the CDR3-LDP and CDR3-LDP-CDR3 fusion proteins demonstrated high affinity towards antigen gelatinases. Following stimulation of CDR3-LDP with enediyne, the results of MTT showed potent cytotoxicity towards tumor cells; the IC50 values of CDR3-LDP-AE to HepG2 and Bel-7402 tumor cells were 1.05x10-11 and 6.6x10-14 M, respectively. In addition, CDR3-LDP-AE displayed a potent antitumor effect in H22 cell xenografts in mice; the combination of CDR3-LDP (10 mg/kg) and CDR3-LDP-AE (0.25 and 0.5 mg/kg) revealed that the tumor inhibitory rates were 85.2 and 92.7%, respectively (P<0.05 compared with CDR3-LDP-AE). In conclusion, these results suggest that the CDR3-LDP fusion protein and its analog CDR3-LDP-AE may both be promising candidates for tumor targeting therapy.

View Figures

View References

1	Björklund M and Koivunen E: Gelatinase-mediated migration and invasion of cancer cells. Biochim Biophys Acta. 1755:37–69. 2005.PubMed/NCBI
2	Kallakury BV, Karikehalli S, Haholu A, Sheehan CE, Azumi N and Ross JS: Increased expression of matrix metalloproteinases 2 and 9 and tissue inhibitors of metalloproteinases 1 and 2 correlate with poor prognostic variables in renal cell carcinoma. Clin Cancer Res. 7:3113–3119. 2001.
3	Svagzdys S, Lesauskaite V, Pangonyte D, Saladzinskas Z, Tamelis A and Pavalkis D: Matrix metalloproteinase-9 is prognostic marker to predict survival of patients who underwent surgery due to rectal carcinoma. Tohoku J Exp Med. 223:67–73. 2011. View Article : Google Scholar : PubMed/NCBI
4	Kessenbrock K, Plaks V and Werb Z: Matrix metalloproteinases: regulators of the tumor microenvironment. Cell. 141:52–67. 2010. View Article : Google Scholar
5	Escaff S, Fernández JM, González LO, et al: Comparative study of stromal metalloproteases expression in patients with benign hyperplasia and prostate cancer. J Cancer Res Clin Oncol. 137:551–555. 2011. View Article : Google Scholar : PubMed/NCBI
6	Shao RG and Zhen YS: Enediyne anticancer antibiotic lidamycin: chemistry, biology and pharmacology. Anticancer Agents Med Chem. 8:121–131. 2008.
7	Guo XF, Zhu XF, Shang Y, Zhang SH and Zhen YS: A bispecific enediyne-energized fusion protein containing ligand-based and antibody-based oligopeptides against epidermal growth factor receptor and human epidermal growth factor receptor 2 shows potent antitumor activity. Clin Cancer Res. 16:2085–2094. 2010. View Article : Google Scholar
8	Xin C, Ye S, Ming Y, et al: Efficient inhibition of B-cell lymphoma xenografts with a novel recombinant fusion protein: anti-CD20Fab-LDM. Gene Ther. 17:1234–1243. 2010. View Article : Google Scholar : PubMed/NCBI
9	Zhong GS, Wu MN, Guo XF, Zhang SH, Miao QF and Zhen YS: Antitumor activities of dFv-LDP-AE: An enediyne-energized fusion protein targeting tumor-associated antigen gelatinases. Oncol Rep. 28:1193–1199. 2012.
10	Zhong G, Zhang S, Li Y, Liu X, Gao R, Miao Q and Zhen Y: A tandem scFv-based fusion protein and its enediyne-energized analogue show intensified therapeutic efficacy against lung carcinoma xenograft in athymic mice. Cancer Lett. 295:124–133. 2010. View Article : Google Scholar
11	Lowe DC, Gerhardt S, Ward A, et al: Engineering a high-affinity anti-IL-15 antibody: crystal structure reveals an α-helix in VH CDR3 as key component of paratope. J Mol Biol. 406:160–175. 2011.PubMed/NCBI
12	Qiu XQ, Wang H, Cai B, Wang LL and Yue ST: Small antibody mimetics comprising two complementarity-determining regions and a framework region for tumor targeting. Nat Biotechnol. 25:921–929. 2007. View Article : Google Scholar : PubMed/NCBI
13	Guo XF, Zhong GS, Miao QF and Zhen YS: Construction of energized fusion protein consisting of epidermal growth factor receptor oligopeptide ligand and lidamycin and its antitumor activity. Ai Zheng. 28:561–568. 2009.(In Chinese).
14	Miao QF, Shang B, Ouyang Z, Liu X and Zhen Y: Generation and antitumor effects of an engineered and energized fusion protein VL-LDP-AE composed of single-domain antibody and lidamycin. Sci China C Life Sci. 50:447–456. 2007. View Article : Google Scholar : PubMed/NCBI
15	Shao RG and Zhen YS: Relationship between the molecular composition of C1027, a new macromolecular antibiotic with enediyne chromophore, and its antitumor activity. Yao Xue Xue Bao. 30:336–342. 1995.(In Chinese).
16	Ashish B and Murthy GS: Analysis of human chorionic gonadotropin-monoclonal antibody interaction in BIAcore. J Biosci. 29:57–66. 2004. View Article : Google Scholar : PubMed/NCBI
17	Quinn JG and O’Kennedy R: Biosensor-based estimation of kinetic and equilibrium constants. Anal Biochem. 290:36–46. 2001. View Article : Google Scholar : PubMed/NCBI
18	Ru Q, Shang BY, Miao QF, Li L, Wu SY, Gao RJ and Zhen YS: A cell penetrating peptide-integrated and enediyne-energized fusion protein shows potent antitumor activity. Eur J Pharm Sci. 47:781–789. 2012. View Article : Google Scholar : PubMed/NCBI
19	Adams GP, Schier R, McCall AM, et al: High affinity restricts the localization and tumor penetration of single-chain Fv antibody molecules. Cancer Res. 61:4750–4755. 2001.PubMed/NCBI
20	Carter PJ: Potent antibody therapeutics by design. Nat Rev Immunol. 6:343–357. 2006. View Article : Google Scholar : PubMed/NCBI

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Small antibody fusion proteins with complementarity‑determining regions and lidamycin for tumor targeting therapy

This article is mentioned in:

Abstract

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Related Articles