Studies On Structure And Function Of Alloferon Derivatives

Alloferon,a thirteen peptides HGVSGHGQHGVHG found from bacterial-infected blow fly C.vicina,was used as anti-pathogen medication with multiple biological functions of anti-cancer(ACP),anti-microbes(AMP),anti-inflammation(AIP)and immune-modulatory activity,which stimulates the cytotoxicity of natural killer cell(NK)and futher regulate the level of interferon(IFN).Alloferon was successfully launched in Russia and ex-Soviet-union(Allokin-alpha)with therapeutic indication for HPV infected cervical cancer since 2003.Alloferon presents itself without the directly therapeutic mechanism imitating in biological function of immunogenicity,cytotoxicity,mutation or carcinogenic effects,embryonic or reproductive toxicity.In view of that great advantages,many researcher pay more attention to the derivatives based on Alloferon,aiming to discover the potential antimicrobial peptide,anticancer peptide and anti-inflammatory peptide,with more effective and safety.According to the papers and peptide database(APD3、CancerPPD),We design the therapeutic and/or preventative peptide library Alloferon-X by chemical or biological modification,including poly ethyl-glycol(PEG),sialic acid linkage...and so on.For theoretical simulation 1)we adopt the bio informatics software or platform,such as ProtParam,APD3,Immunomedicine Group,NPS@,TMHMM to analyse the physicochemical properties,secondary structure,transmembrane ability of Alloferon-X;2)we use the Hyperchem software to construct the Alloferon-Cu(Ⅱ)、Alloferon-Zn(II)complex,to verify the influence of metal ions to Alloferon(ΔE<0);3)we utilize the PeptideQSAR program to model QSAR for analogues of Alloferon,which have been reported by others,to predict the potential activity of peptide(Q2>0.5).For experimental study,we screened the Alloferon-X,choosing the PEG-modification and sialic acid-modification peptides(N-terminal)to carry out anticancer,immune-modulatory,anti-bacterial activity.The study shown that 1)Alloferon and PEG-modification derivatives(0-500 mg/L range)had shown no direct killing effects to Hela,also Alloferon(100μM)had shown no indirect cyctotoxicity effect to PBMC immune-cells&Hela co-culture system(E:T),while sialic acid-Alloferon(100 μM)had shown the cyctotoxicity effect to PBMC immune-cells&Raji,THP-1 co-culture system,with maximam cyctotoxicity effect were 21%,34%resepectively for E:T=1:1;2)PEG2-Alloferon、PEG6-Alloferon(100 μM)had shown the immune-modulatory activity by promoting the level of TNF-α in THP-1 induced macrophage model,while sialic acid-Alloferon did not affect the level of IFN-y in PBMC,NK&Raji co-culture system significantly;3)In KB method,Alloferon(100 mg/mL)had shown no anti-bacterial activity to E.coli.