High throughput screening of 66,000 compounds using competitive binding of peptides comprising the BH3 domain to anti-apoptotic Bfl-1 led to the identification of fourteen validated "hits" as inhibitors of Bfl-1. N-Aryl maleimide 1 was among the validated "hits". A chemical library encompassing over 280 analogs of 1 was prepared following a two-step synthesis. Structure-activity studies for inhibition of Bfl-1 by analogs of N-aryl maleimide 1 revealed a preference for electron-withdrawing substituents in the N-aryl ring and hydrophilic amines appended to the maleimide core. Inhibitors of Bfl-1 are potential development candidates for anticancer therapeutics.
Keywords
Bfl-1 inhibitors; N-aryl maleimides; high-throughput screening; anti-cancer agentsThe B-cell lymphoma/leukemia-2 (Bcl-2) family of proteins plays a prominent role in regulating programmed cell death (apoptosis). In humans, proteins within the Bcl-2 family having anti-apoptotic activity include Bcl-2, Bcl-X L , Mcl-1, Bcl-W, Bcl-B, and Bfl-1. 1 Members of this group of proteins are commonly over-expressed in human cancers thereby promoting tumorigenesis. 2,3 Additional groups of proteins within the Bcl-2 family have pro-apoptotic activity and include Bak, Bax, Bad, Bim, and Bid. Anti-apoptotic and proapoptotic proteins can dimerize, either activating or antagonizing each other. 4 Heterodimerization occurs at a hydrophobic pocket on the surface of anti-apoptotic proteins and a region of pro-apoptotic proteins called the BH3 region to neutralize the cytoprotective function of anti-apoptotic Bcl-2 family proteins. The BH3 region is comprised of an amphipathic α-helix of roughly 16-25 amino acids that binds to the hydrophobic region of the anti-apoptotic Bcl-2 family proteins. 1 Agents that mimic the BH3 region of proapoptotic proteins may be used to antagonize anti-apoptotic Bcl-2 proteins by binding to the hydrophobic pocket thus promoting cell death. Synthetic peptides mimicking the BH3 region have been shown to bind to the anti-apoptotic Bcl-2 proteins and initiate apoptosis. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Bfl-1 is a NF-κB-inducible member of the Bcl-2 family that is highly expressed in immune cells 15 . Defective apoptosis has been implicated in several autoimmune diseases, thus suggesting a therapeutic opportunity for agents that neutralize Bfl-1. In addition, Bfl-1 is highly expressed in some cancers accounting for resistance to selective Bcl-2/Bcl-X L inhibitors that fail to neutralize this anti-apoptosis member of the Bcl-2 family. Based on these results, we decided to focus on sm...
HIV infection of a fetus from an infected mother has severe immunological implications because it destroys still immature immune system. The abnormalities observed are precocious in the infants. When the infection occurs in perinatal period, clinical and immunological manifestations can present themselves at the age of 2-5 years. Two patterns of HIV infection can be distinguished. Humoral immunodeficiency is present in a high proportion of patients and leads to repeated bacterial infections and progression of the disease. Immunoglobulin substitution therapy improves clinical manifestations and can help to avoid viral replication.
Several approaches known for producing "neometalloenzymes" are classified into two categories: protein engineering using antibodies as starting materials and "de novo" biosynthesis of metal-binding antibodies with potential catalytic metal-binding structure. This latter approach is chosen in this study. Polyclonal anti-zinc-iminodiacetate [IDA-Zn(II)] antibodies are produced in rabbits and mice. Because of the absolute need for the unequivocal screening of the hapten [IDA-Zn(II)] specific antibodies, a new ELISA method was developed using a biheaded polyethylene glycol with biotin on one end and the hapten on the other end. The parameters for optimizing the immunization and the ELISA technique are discussed and the method is validated with rabbit and mice sera.
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