Edb targeting il-12 compositions
A technology of IL-12 and composition, which is applied in the field of IL-12 composition targeting EDB, can solve problems such as hindering the development of anticancer drugs, and achieve the effect of improving therapeutic potential and excellent manufacturability
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Embodiment 1
[0272] Applicants have surprisingly found that when certain linkers are used to link IL-12 to a single chain diabody (i.e. the superior form disclosed in WO2013 / 014149), better tumor targeting performance as well as superior production output.
[0273] Applicants have evaluated and characterized eight cloned single-chain diabody forms of human IL-12 (huIL-12L19L19) linked to L19 antibodies with different polypeptide linkers between the cytokine and the L19 single-chain diabody.
[0274] Five clones (designated: (i) "AKKAS", (ii) "DDS", (iii) "(G4S) 3 ", (iv) "SAD" and (v) "SES") contain linkers for conjugation of immunocytokine to recombinant antibody and were selected for their different charge characteristics (neutral, positively charged, negatively charged) .
[0275]Three other clones were developed, named (vi) α3, (vii) AP6 and (viii) AP7. For these three clones, the teaching reported in Chen et al. (2013) was considered and implemented. This review suggests that rigi...
Embodiment 2
[0316] In another set of comparative experiments, it was surprisingly found that the "SAD" linker is also superior to the old (and shorter) GSADGG linker disclosed in WO2013 / 014149 in terms of binding ability, monomer profile and tumor targeting ability ( SEQ ID NO 26).
[0317] Materials and methods
[0318]The variants tested in this example have the following common structure:
[0319]
[0320] The different variants (also referred to herein as "clones") differ from each other with respect to the sequence of linker 2:
[0321] Connector 2 SEQ ID NO sequence SAD 4 GSADGGSSAGGSDAG old 26 GSADGG
[0322] Cloning of fusion proteins
[0323] Fusion proteins were cloned along the lines described above, comprising huIL-12 fused to the L19 antibody via a 6 or 15 amino acid linker, in the form of a single-chain diabody (i.e., huIL-12L19L19 "Old" and huIL-12L19L19, respectively "SAD" variant).
[0324] Expression of fusion protein
[0325] I...
Embodiment 3
[0344] Evaluating the efficacy of huIL-12L19L19 "SAD variants" in human patients with malignant melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma, urothelial carcinoma, squamous cell carcinoma of the head and neck Carcinoma (HNSCC), microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR) metastatic colorectal cancer, hepatocellular carcinoma, gastric cancer, squamous cell carcinoma of the skin, cervical cancer, and diffuse large B cell lymphoma (DLBCL). At least one cohort of patients exhibited disease progression as part of an immune checkpoint blockade-based regimen administered with the most recent prior therapy.
[0345] Patients received the huIL-12L19L19 "SAD" variant by intravenous administration once a week for 8 weeks. Patients received doses of 4 μg / kg; 8 μg / kg; 12 μg / kg; 16 μg / kg; or 20 μg / kg.
[0346] Patients were followed for 6 months from the start of treatment, or until withdrawal of consent or until progressive disease. ...
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