Invented by Chi-Huey Wong, Tsui-Ling Hsu, Yi-Wei Lou, Chih-Wei Lin, Shih-Chi Yeh, Chung-Yi Wu, Han-Chung Wu, Academia Sinica
The Academia Sinica invention works as followsThe present invention discloses a pharmaceutical composition containing antibodies or antigen-binding fragments of antibodies that bind to SSEA3, SSEA4, and globo H. It also describes methods for using the composition. The methods of use are not limited to cancer diagnostics and therapies. The antibodies disclosed herein can bind certain cancer cell surface. The antibodies can be used to target carcinomas in the brain, skin and bone, as well as cancers of the lungs, esophagus or stomach, liver, gallbladder, pancreas.
Background for Compositions for the treatment and detection cancer
Glioblastoma Multiforme (GBM) is the most aggressive form and most common primary brain tumour in adults. (Louis D. N., et. al. The 2007 WHO classification for tumours in the central nervous systems. Acta Neuropathol 114(2):97-109). The prognosis for GBM is poor despite the availability of many treatments including surgery, radiotherapy, and chemotherapy. The median survival rate is 14-15 months. 2010 Exciting new advancements in neuro-oncology : The avenue to a cure of malignant glioma. CA Cancer J Clin 60(Meyer M A (2008) Malignant gliomas in adults. N Engl J Medicine 359(17), 1850 (author reply, 1850) 166-193). GBMs are notoriously resistant to anti-cancer medications and infiltrative. This makes it difficult to perform a complete surgical resection. Most patients experience tumor progression or recurrence even after multiple treatments. Due to the high mortality rate, new approaches have been suggested for treating GBM, including immunotherapy and genetic therapy (Meyer, M. A. (2008) Malignant Gliomas in Adults). N Engl J Medicine 359(17), 1850, author’s reply (1850).
Altered glycosylation is a feature of cancer cells, and several glycan structures are well-known tumor markers (Meezan E, Wu H C, Black P H, & Robbins P W (1969) Comparative studies on the carbohydrate-containing membrane components of normal and virus-transformed mouse fibroblasts. II. Separation by sephadex of glycoproteins from glycopeptides. Hakomori, S. (2002). Glycosylation as a marker of cancer malignancy. A new wine in a old bottle. Proc Natl Acad Sci USA 99(16):10231-10233). This aberrant change includes the increase in N-linked glycan branching (Lau K S and Dennis J W (2008) N’Glycans as a cancer progression. Glycobiology (18(10):750-760)) and sialic content (van Beek, W. P., Smets, L. A., and Emmelot, P. (1973) Increased density of sialic acids in surface glycoproteins of malignant and transformed cells?A general phenomenon? Cancer Res 33(11), 2913-2922), as well as the overexpression and glycan epitopes such sialyl Lewis (sLex), Lewis (Ley), Globo (H), and polysialic acids (Sell (1990), Cancer-associated carbohydrates detected by monoclonal antibody. Hakomori S. & Zhang Y. (1997) Glycosphingolipid Antigens and Cancer Therapy. Chem Biol, 4(2):97 – 104. Taylor-Papadimitriou J & Epenetos AA (1994) Exploiting changed glycosylation patterns to diagnose and treat cancer. Trends Biotechnol, 12(6):227-233. Several tumors show increased expression of glycolipids. These include gangliosides and glycosphingolipids with sialic acids attached to the glycan chains. Gangliosides can be found in the neural system and in tumors. This is especially true for complex gangliosides that are associated with cancer (Birkle, Zeng, Gao, Yu, R.K., & Aubry, 2003, Role of tumorassociated gangliosides). Biochimie 85(3-4):455-463).
It was reported that a human glioma showed expression of ganglosides” (Fredman P., et. al. (1986) Potential antigens of gangliosides in human gliomas. Yates A. J., Becker, L. E., & Sachs, L. A. (1979), Childhood brain tumors. Childs Brain 5(1) 31-39. Traylor T D and Hogan E. L. (1980). Gangliosides in human cerebral astrocytomas. J Neurochem, 34(1), 126-131. Berra B., Gaini S. M., & Riboni, L. (1985). Correlation of gangliosides distribution with histological grade in human astrocytomas. Int J Cancer 36(3):363-366; Fredman P, von Holst H, Collins V P, Granholm L, & Svennerholm L (1988) Sialyllactotetraosylceramide, a ganglioside marker for human malignant gliomas. J Neurochem 50(3):912-919; Mansson J E, et al. (1986) Characterization and characterization of lactotetraose gangliosides in murine xenografts derived from a human glioma. FEBS Lett 201(1) : 109-113. Fredman P., von Holst, Collins V P., Dellheden, B., & Svennerholm, L. (1993). Expression of gangliosides GD3 & 3?-isoLM1 on autopsy brains taken from malignant tumor patients. J Neurochem 60(1):99-105). Some glioma associated gangliosides may be absent or only rarely expressed in normal tissues. (Svennerholm, L. et al. Human brain gangliosides (1989): Developmental changes from the early fetal to advanced age. Biochim Biophys Acta 1005(2): 109-117) and are therefore suitable for targeted treatment (Kato, et. al. (2010) GMab-1, a high-affinity anti-3?-isoLM1/3?,6?-isoLD1 IgG monoclonal antibody, raised in lacto-series ganglioside-defective knockout mice. Biochem Biophys Res Commun 391(1):750-755). Discovering novel GSLs associated with gliomas would therefore provide new targets for the development of new therapies to treat gliomas.
The GSLs of globo-series feature a Gal?1-4Gal linkage to lactosylceramides, and this linkage is catalyzed by lactosylceramide 4-alpha-galactosyltransferase (A4GALT). The P-blood system is based on globosides (Gb4Cer), globotriosylceramides (Gb3Cer), and globotriosylceramides (Gb3Cer). Human Blood Groups (Springer Vienna), pages 211-234. Galactosyl-globosides (Gb5Cer), and sialyl-galactosyl-globosides (sialyl-Gb5Cer), are also called stage-specific embryonic antibodies 3 (SSEA-3), and 4 (SSEA-4). Stage-specific embryonic antibodies (SSEA-3 & -4) were isolated in 1983 from human teratocarcinoma cell epitopes. EMBO J (2(12):2355-2361) are used to identify human embryonic stem cell. Globo series GSLs are also observed in tumors. Globo H, fucosyl Gb5Cer, is overexpressed by many epithelial carcinomas such as ovarian cancer, gastric cancer, prostate cancer, lung cancer, breast cancer, and pancreatic (Zhang S, etc.). (1997) Selection of tumor antigens as targets for immune attack using immunohistochemistry: I. Focus on gangliosides. Int J Cancer, 73(1), 42-49; SSEA-3 and SSEA-4, as well as Globo H, are expressed on both breast cancer cells and breast cancer stem cells. (Chang, W. W., et. al. Globo H, SSEA3 and fucosyltransferases 1 and 2, which are involved in Globo H synthesis in breast cancer stem cell expression were studied in 2008. Proc Natl Acad Sci USA 105(33):11667-11672; Huang Y L, et al. Carbohydrate based vaccines for breast cancer with glycolipid adjuvants. Proc Natl Acad Sci USA 110(7):2517-2522). In addition, the expression of SSEA-4 (disialosyl gb5cer, DSGG), and disialosyl globoside is high in renal cell carcinoma (Saito S. et. al. (1997) Expression globo series gangliosides of human renal cell carcinoma. Jpn J Cancer Res. 88(7):652-659). However, it is not yet known whether GSLs of the globo series are expressed in GBM.
It is important to identify glycans that are associated with or predictive of cancers and develop antibodies to the markers in order for them be used as diagnostic and treatment tools.
The present disclosure is based also on the discovery of Globo H expression in cancer cells, but not normal cells. Globo-expressing cancer cells include brain cancers, lung cancers, breast cancers, oral cancers, esophageal and stomach cancers, liver and bile ducts cancers, skin cancers, prostate and ovarian tumors, and pancreatic and colon cancers.
The disclosure, in one aspect provides affinity-matured SSEA-3/SSEA-4/Globo-H antibodies. These antibodies are more specific and have a higher affinity for SSEA-3, SSEA-4/Globo H.
In one embodiment, SSEA-3/SSEA-4/Globo-H antibodies are useful in treating SSEA-3/SSEA-4/Globo-H-mediated disorders where a partial or complete blockade is desired of one or more SSEA-3/SSEA-4/Globo-H activities. The anti SSEA-3, SSEA-4, and Globo H antibodies are used in one embodiment to treat cancer.
The anti-SSEA-3/SSEA-4/Globo H antibodies of the invention permit the sensitive and specific detection of the epitopes in immunoassays, for example, immunoprecipitations, sandwich immunoassays, ELISAs, or immunomicroscopy without the need for mass spectrometry or genetic manipulation. This provides an advantage for observing the normal function of these pathways as well as detecting abnormalities.
SSEA4/SSEA3/GloboH are three glycans which are expressed specifically by cancer cells and cancer-stem cells. The knockdown of beta-3 GalT5, which is the enzyme responsible for synthesising these glycolipids in cancer cells but not in normal cells, results in apoptosis. Antibodies, and especially glycoantibodies, against SSEA4 specifically or preferentially, as well as against SSEA3/SSEA4/GloboH simultaneously, are therefore effective cancer therapeutic agents. “In another aspect, SSEA3, SSEA4/SSEA3/GloboH, and especially SSEA3, can be used as cancer stem cell markers.
SSEA4 or SSEA4/SSEA3/GloboH in combination is useful as a therapeutic target for different cancers. For example, brain, lung, breast, oral, esophageal, liver, bile-duct, skin, pancreatic, colon, kidney, and prostate cancers.
In one embodiment, humanized or humanized therapeutic antibody against SSEA4 is expressed on the surface of these cancer types.
In another embodiment, humanized or humanized therapeutic antibody against SSEA3/SSEA4/GloboH simultaneously expressed on the surface of these exemplary types of cancer are provided.
The present disclosure includes an isolated SSEA-4 antibody. The anti-SSEA-4 antibody binds to Neu5Ac?2?3Gal?1?3GalNAc?1?3Gal?1?4Gal?1?4Glc?1 (SSEA-4 hexasaccharide). In some examples, the antibody is capable of binding to Neu5Gc?2?3Gal?1?3GalNAc?1?3Gal?1?4Gal?1?4Glc?1 (an analogue of SSEA-4 hexasaccharide).
The present disclosure also includes an isolated antibody that is specific for SSEA-4 or fragments of SSEA-4. The anti-SSEA-4 antibody binds to Neu5Ac?2?3Gal?1?3GalNAc?1?3Gal?1?4Gal?1?4Glc?1 (SSEA-4 hexasaccharide) and Neu5Ac?2?3Gal?1?3GalNAc?1?3Gal?1 (fragment of SSEA-4 hexasaccharide). In some examples, the antibody is capable of binding to Neu5Ac?2?3Gal?1?3GalNAc?1?3Gal?1. In some examples, the antibody is capable of binding to Neu5Ac?2?3Gal?1?3GalNAc?1?3Gal?1?4Gal?1?4Glc?1 (an analogue of SSEA-4 hexasaccharide).
The present disclosure, in one aspect, provides an isolated antigen-binding antibody or fragment, which includes H-CDR1,H-CDR2,and H-CDR3, selected from (i), (ii), as shown in Table 17-4. 17):
(i) Select H-CDR1 from SEQ ID No:152
(iii) H CDR2 selected from SEQ No:153
(iii), H-CDR3 is selected from SEQ NO: 154 respectively.
and including L-CDR1, L CDR2, and L CDR3 selected from (iv), to (vi)
(iv), L-CDR1 from SEQ ID No: 149 selected;
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