Synthesis and Evaluation of the Aldolase Antibody-Derived Chemical-Antibodies Targeting α5β1 Integrin

Abstract

Integrin α5β1 is an important therapeutic target that can be inhibited using an aldolase antibody (Ab)-derived chemical-Ab (chem-Ab) for the treatment of multiple human diseases, including cancers. A fairly optimized anti-integrin α5β1 chem-Ab 38C2-4e was obtained using an in situ convergent chemical programming (CP) approach, which minimized the time and efforts needed to develop a chem-Ab. Multiple Ab-programming agents (PAs) 4a-e could be prepared rapidly using the Cu-catalyzed alkyne-azide coupling (Cu-AAC) reaction of an α5β1 inhibitor 2 with multiple linkers 3a-e, either before or after conjugating the linkers into Ab 38C2 binding sites. In these two-steps processes, the products after step 1 can be used in next step without performing an extensive purification or analysis of the Ab-PAs or Ab-linker conjugates affording chem-Abs 38C2-(4a-e). Flow cytometry assay was used to determine binding of the chem-Abs to U87 human glioblastoma cells expressing α5β1 integrin, and identify 38C2-3e as the strongest binder. Further studies revealed that 38C2-3e strongly inhibited proliferation of U87 cells and tube formation of HUVEC in matrigel assay, as well as tumor growth and metastasis of 4T1 cells in vivo.

Chemically Programmed Bispecific Antibody Targeting Legumain Protease and αvβ3 Integrin Mediates Strong Antitumor Effects

“…In contrast, fewer U87 cells formed spheroids in control (Ab 38C2) and 38C2- 3 treated groups. The results were similar to those observed with the integrin α5β1 inhibitors and anti-α5β1 cPAbs …”

“…The results were similar to those observed with the integrin α5β1 inhibitors and anti-α5β1 cPAbs. 33 Development of functional vasculatures requires precise spatial-temporal regulation of endothelial cell proliferation and invasion. To determine and analyze the effects that can be engendered by using the anti-legumain, anti-integrin cp-bsAb 38C2-5 on tumor vascularization, human endothelial cell functions were assessed in vitro in Matrigel using the tube formation assay, as described in the literature.…”

Clickable Poly(ethylene glycol)‐Based Copolymers Using Azide–Alkyne Click Cycloaddition‐Mediated Step‐Growth Polymerization

“…Sodium hydride, propargyl bromide, Cu(I) bromide, Cu(I) iodide, bromotris(triphenylphosphine)copper(I), N,N,N ′, N ′, N ′′‐penta­me­thyldiethylenetriamine (PMDTA), 1,8‐diazabicyclo[5.4.0]‐ undec‐7‐ene (DBU), N,N ‐Diisopropylethylamine (DIPEA), benzyl mercaptan, 2,2‐dimethoxy‐2‐phenylacetophenone (DMPA), 3,6‐di‐2‐pyridyl‐1,2,4,5‐tetrazine, anthracene, and 2,2′‐(ethylene­dioxy)diethanethiol were obtained from Aldrich. Homobifunctional TEG monomers with alkyne (diyne‐TEG) and azide (diazide‐TEG) groups at their termini were synthesized according to reported procedures and confirmed with a spectroscopic analysis. Compound diyne‐FuMAL was synthesized according to previously published procedure …”

Shedding New Light on The Role of ανβ3 and α5β1 Integrins in Rheumatoid Arthritis

“…It has the ability to inhibit α5/fibronectin interaction. During preclinical studies, volociximab induced in vitro and in vivo endothelial cell apoptosis and prevented blood vessel formation [87]. Anti-angiogenic and anti-tumor activities were revealed in chick chorioallantoic membrane (CAM) following volociximab inoculation.…”