Monomethyl auristatin E-conjugated anti-EGFR antibody inhibits the growth of human EGFR-positive non-small cell lung cancer
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Epidermal growth factor receptor (EGFR) is highly expressed on non-small cell lung cancers (NSCLC) and a valuable therapeutic target. This study aimed at producing and characterizing monomethyl auristatin E (MMAE)-conjugated anti-EGFR antibody as a novel EGFR-targeting therapy for NSCLC.
A humanized anti-EGFR monoclonal antibody (named RC68) was purified and conjugated with MMAE using a MC-VC-PAB or PY-VC-PAB linker. The in vitro and in vivo antitumor activity of RC68-MC-VC-PAB-MMAE and RC68-PY-VC-PAB-MMAE were characterized.
The RC68 was generated from RC68-expressing cells and had a purity of > 99.0%. The RC68 recognized EGFR on tumor cells, particularly for higher EGFR expressing H125, A431, HCC827 and H1975 cells. The RC68 was conjugated with an average of 4 MMAE molecules to generate RC68-MC-VC-PAB-MMAE and RC68-PY-VC-PAB-MMAE, respectively. The RC68-MC-VC-PAB-MMAE, RC68-PY-VC-PAB-MMAE and RC68 displayed similar binding affinity to EGFR on tumor cells, and RC68-MC-VC-PAB-MMAE and RC68-PY-VC-PAB-MMAE were effectively internalized by H125 cells. The RC68-MC-VC-PAB-MMAE and RC68-PY-VC-PAB-MMAE inhibited the growth of H125 cells in vitro with an IC50 7.37–8.04 ng/mL and implanted H125 tumors in vivo, but did not affect body weights of mice. The antitumor effect of RC68-MC-VC-PAB-MMAE was stronger than RC68-PY-VC-PAB-MMAE, which was also stronger than docetaxel in vivo.
These novel antibody–drug conjugates, particularly for RC68-MC-VC-PAB-MMAE, may be a potential candidate for treatment of EGFR + NSCLC.
KeywordsEpidermal growth factor receptor Non-small cell lung cancer Monoclonal antibody Antibody–drug conjugates
The authors thank both Hongwen Li and Xiaoyu Xu at RemeGen Ltd. for their help in preparing RC68 antibody.
This work was supported by a Grant from the National Science and Technology Major Project of China (Grant number: 2014ZX09508004-003).
Compliance with ethical standards
Conflict of interest
The authors declare no conflicts of interest.
Animal experiment was approved by Medicilon and carried out in accordance with the institutional guidelines. This study did not contain any experiment with human participants.
- 1.Schrank Z, Chhabra G, Lin L et al (2018) Current molecular-targeted therapies in NSCLC and their mechanism of resistance. Cancers (Basel) 10(7):1–17Google Scholar
- 2.Fuster LM, Sandler AB (2004) Select clinical trials of erlotinib (OSI-774) in non-small cell lung cancer with emphasis on phase III outcomes. Clin Lung Cancer 6(Suppl 1):S24–29Google Scholar
- 3.Spira A, Ettinger DS (2004) Multidisciplinary management of lung cancer. N Engl J Med 350(4):379–392Google Scholar
- 4.Gadgeel SM, Ramalingam SS, Kalekerian GP (2012) Treatment of lung cancer. Radiol Clin N Am 50(5):961–974Google Scholar
- 5.Yarden Y, Sliwkowski MX (2001) Untangling the ErbB signaling network. Nat Rev Mol Cell Biol 2(2):127–137Google Scholar
- 6.Mendelsohn J, Baselga J (2006) Epidermal growth factor receptor targeting in cancer. Semin Oncol 33(4):369–385Google Scholar
- 7.Jorissen RN, Walker F, Pouliot N et al (2003) Epidermal growth factor receptor: mechanisms of activation and signaling. Exp Cell Res 284(1):31–53Google Scholar
- 8.Domvri K, Zarogoulidis P, Darwiche K et al (2013) Molecular targeted drugs and biomarkers in NSCLC, the evolving role of individualized therapy. J Cancer 4(9):736–754Google Scholar
- 9.Shaw RJ, Cantley LC (2006) Ras, PI(3)K and mTOR signalling controls tumor cell growth. Nature 441(7092):424–430Google Scholar
- 10.Ohsaki Y, Tanno S, Fujita Y et al (2000) Epidermal growth factor receptor expression correlates with poor prognosis in non-small cell lung cancer patients with p53 overexpression. Oncol Rep 7(3):603–607Google Scholar
- 11.Mendelsohn J, Baselga J (2003) Status of epidermal growth factor receptor antagonists in the biology and treatment of cancer. J Clin Oncol 21(14):2787–2799Google Scholar
- 12.Enrique AA, Gema PC, Jeronimo JC et al (2012) Role of anti-EGFR target therapy in colorectal carcinoma. Front Biosci (Elite Ed) 4:12–22Google Scholar
- 13.Moran T, Sequist LV (2012) Timing of epidermal growth factor receptor tyrosine kinase inhibitor therapy in patients with lung cancer with EGFR mutations. J Clin Oncol 30(27):3330–3336Google Scholar
- 14.Barker AJ, Gibson KH, Grundy W et al (2001) Studies leading to the identification of ZD1839 (Iressa): an orally active, selective epidermal growth factor receptor tyrosine kinase inhibitor targeted to the treatment of cancer. Bioorg Med Chem Lett 11(14):1911–1914Google Scholar
- 15.Moyer JD, Barbacci EG, Iwata KK et al (1997) Induction of apoptosis and cell cycle arrest by CP-358,774, an inhibitor of epidermal growth factor receptor tyrosine kinase. Cancer Res 57(21):4838–4848Google Scholar
- 16.Li D, Ambrogio L, Shimamura T et al (2008) BIBW2992, an irreversible EGFR/HER2 inhibitor highly effective in preclinical lung cancer models. Oncogene 27(34):4702–4711Google Scholar
- 17.Greig SL (2016) Osimertinib: first global approval. Drugs 76(2):263–273Google Scholar
- 18.Pao W, Miller VA, Politi KA et al (2005) Acquired resistance of lung adenocarcinomas to gefitinib or erlotinib is associated with a second mutation in the EGFR kinase domain. PLoS Med 2(3):225–235Google Scholar
- 19.Yun CH, Mengwasser KE, Toms AV et al (2008) The T790M mutation in EGFR kinase causes drug resistance by increasing the affinity for ATP. Proc Natl Acad Sci USA 105:2070–2075Google Scholar
- 20.Yu HA, Sima CS, Huanq J et al (2013) Local therapy with continued egfr tyrosine kinase inhibitor therapy as a treatment strategy in egfr-mutant advanced lung cancers that have developed acquired resistance to egfr tyrosine kinase inhibitors. J Thorac Oncol 8(3):346–351Google Scholar
- 21.Losanno T, Rossi A, Maione P et al (2016) Anti-EGFR and antiangiogenic monoclonal antibodies in metastatic non-small-cell lung cancer. Expert Opin Biol Ther 16(6):747–758Google Scholar
- 22.Bakema JE, van Egmond M (2014) Fc receptor-dependent mechanisms of monoclonal antibody therapy of cancer. Curr Top Microbiol Immunol 382:373–392Google Scholar
- 23.di Noia V, D'Argento E, Pilotto S et al (2018) Necitumumab in the treatment of non-small-cell lung cancer: clinical controversies. Expert Opin Biol Ther 18(9):937–945Google Scholar
- 24.Peters C, Brown S (2015) Antibody–drug conjugates as novel anti-cancer chemotherapeutics. Biosci Rep 35(4):1–20Google Scholar
- 25.Lambert JM, Berkenblit A (2018) Antibody–drug conjugates for cancer treatment. Annu Rev Med 69:191–207Google Scholar
- 26.van den Bent M, Gan HK, Lassman AB et al (2017) Efficacy of depatuxizumab mafodotin (ABT-414) monotherapy in patients with EGFR-amplified, recurrent glioblastoma: results from a multi-center, international study. Cancer Chemother Pharmacol 80(6):1209–1217Google Scholar
- 27.Jo U, Park KH, Whang YM et al (2014) EGFR endocytosis is a novel therapeutic target in lung cancer with wild-type EGFR. Oncotarget 5(5):1265–1278Google Scholar
- 28.Li Z, Wang M, Yao X et al (2019) Development of a novel EGFR-targeting antibody–drug conjugate for pancreatic cancer therapy. Target Oncol 14(1):93–105Google Scholar
- 29.Yao X, Jiang J, Wang X et al (2015) A novel humanized anti-HER2 antibody conjugated with MMAE exerts potent antitumor activity. Breast Cancer Res Treat 153(1):123–133Google Scholar
- 30.Huang C, Fang J, Ye H et al (2017) Covalent linkers in antibody–drug conjugates and methods of making and using the same [P]. WO 2017/031034 A2Google Scholar
- 31.Tumey LN, Charati M, He T et al (2014) Mild method for succinimide hydrolysis on ADCs: impact on ADC potency, stability, exposure, and efficacy. Bioconjug Chem 25(10):1871–1880Google Scholar
- 32.Riedl T, van Boxtel E, Bosch M et al (2016) High-throughput screening for internalizing antibodies by homogeneous fluorescence imaging of a pH-activated probe. J Biomol Screen 21(1):12–23Google Scholar
- 33.Chen YM (2015) Update of epidermal growth factor receptor-tyrosine kinase inhibitors in non-small-cell lung cancer. J Chin Med Assoc 76(5):249–257Google Scholar
- 34.Stewart EL, Tan SZ, Liu G et al (2015) Known and putative mechanisms of resistance to EGFR targeted therapies in NSCLC patients with EGFR mutations—a review. Transl Lung Cancer Res 4(1):67–81Google Scholar
- 35.Dosio F, Brusa P, Cattel L (2011) Immunotoxins and anticancer drug conjugate assemblies: the role of the linkage between components. Toxins (Basel) 3:848–883Google Scholar
- 36.Tsuchikama K, An Z (2018) Antibody–drug conjugates: recent advances in conjugation and linker chemistries. Protein Cell 9(1):33–46Google Scholar
- 37.Donaghy H (2016) Effects of antibody, drug and linker on the preclinical and clinical toxicities of antibody–drug conjugates. MAbs 8(4):659–671Google Scholar
- 38.Eaton JS, Miller PE, Mannis MJ et al (2015) Ocular adverse events associated with antibody–drug conjugates in human clinical trials. J Ocul Pharmacol Ther 31(10):589–604Google Scholar
- 39.Phillips AC, Boghaert ER, Vaidya KS et al (2018) Characterization of ABBV-221, a tumor-selective EGFR targeting antibody drug conjugate. Mol Cancer Ther 17(4):795–805Google Scholar
- 40.Singh SK, Luisi DL, Pak RH (2015) Antibody–drug conjugates: design formulation and physicochemical stability. Pharm Res 32(11):3541–3571Google Scholar
- 41.McCombs JR, Owen SC (2015) Antibody drug conjugates: design and selection of linker, payload and conjugation chemistry. AAPS J 17(2):339–351Google Scholar
- 42.Hamblett KJ, Kozlosky CJ, Siu S et al (2015) AMG 595, an anti-EGFRvIII antibody–drug conjugate, induces potent antitumor activity against EGFRvIII-expressing glioblastoma. Mol Cancer Ther 14(7):1614–1624Google Scholar