Preparation of an antitumor and antivirus agent: chemical modification of α-MMC and MAP30 from Momordica Charantia L. with covalent conjugation of polyethyelene glycol

Abstract

Background: Alpha-momorcharin (α-MMC) and momordica anti-HIV protein (MAP30) derived from Momordica charantia L. have been confirmed to possess antitumor and antivirus activities due to their RNA-N-glycosidase activity. However, strong immunogenicity and short plasma half-life limit their clinical application. To solve this problem, the two proteins were modified with (mPEG) 2 -Lys-NHS (20 kDa). Methodology/principal findings: In this article, a novel purification strategy for the two main type I ribosome-inactivating proteins (RIPs), α-MMC and MAP30, was successfully developed for laboratory-scale preparation. Using this dramatic method, 200 mg of α-MMC and about 120 mg of MAP30 was obtained in only one purification process from 200 g of Momordica charantia seeds. The homogeneity and some other properties of the two proteins were assessed by gradient SDS-PAGE, electrospray ionization quadruple mass spectrometry, and N-terminal sequence analysis as well as Western blot. Two polyethylene glycol (PEG)ylated proteins were synthesized and purified. Homogeneous mono-, di-, or tri-PEGylated proteins were characterized by matrix-assisted laser desorption ionization-time of flight mass spectrometry. The analysis of antitumor and antivirus activities indicated that the serial PEGylated RIPs preserved moderate activities on JAR choriocarcinoma cells and herpes simplex virus-1. Furthermore, both PEGylated proteins showed about 60%-70% antitumor and antivirus activities, and at the same time decreased 50%-70% immunogenicity when compared with their unmodified counterparts. Conclusion/significance: α-MMC and MAP30 obtained from this novel purification strategy can meet the requirement of a large amount of samples for research. Their chemical modification can solve the problem of strong immunogenicity and meanwhile preserve moderate activities. All these findings suggest the potential application of PEGylated α-MMC and PEGylated MAP30 as antitumor and antivirus agents. According to these results, PEGylated RIPs can be constructed with nanomaterials to be a targeting drug that can further decrease immunogenicity and side effects. Through nanotechnology we can make them low-release drugs, which can further prolong their half-life period in the human body.

Bitter melon: a panacea for inflammation and cancer

“…Meng et al . have developed PEGylated derivatives of α -momocharin and momordica anti-HIV protein (MAP30) which exerts about 60%–70% anti-proliferative in JAR choriocarcinoma cells and anti-viral activities as well as reduces 50%–70% immunogenicity when compared with their unmodified counterparts [87] . Manoharan et al .…”

<p>Silver Nanoparticles for the Therapy of Tuberculosis</p>

“…19 Intramacrophagic killing of M. smegmatis internalized in RAW264.7 macrophages (in both pre/and postexposure treatments) was reported for spherical chitosan-coated AgNP (CS-AgNPs) in 3 ppm dose. 220 The bactericidal effect was time and concentration-dependent and most of the antibacterial effect was observed in the first hour of incubation. The hypothesized antibacterial mechanism was cell membrane disruption or chemical inactivation of thiol-containing molecules.…”

“…In the same study, in addition to antimycobacterial effect, CS-AgNPs were found to be also active against other bacteria like Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella typhi. 220 In addition to the direct antitubercular activity in doses beginning with 5 mg/l, AgNPs measuring 10-150 nm were shown to potentiation of the antibacterial effect of isoniazid, rifampicin, ethionamide, levofloxacin, ofloxacin and kanamycin against clinical isolates of M. tuberculosis by Kreytsberg et al 133 In another study employing the antimycobacterial effect of physicochemically synthesized, tetrahedral and spherical AgNP measuring 50 nm, an antibacterial effect against field isolates and standard strains of M. tuberculosis and M. bovis were reported. The minimal inhibitory concentration (MIC) was found to be 1 and 4 µg/mL for standard cultures of M. tuberculosis and M. bovis.…”

Engineering of Ribosome-inactivating Proteins for Improving Pharmacological Properties

“…Many RIPs PEGylations were attempted to advance their pharmacological properties (Table 1). α-MMC 1 20 kDa (mPEG)2-Lys-NHS Tumor Mono-, di-, tri-PEGylated [112], [113], [114], [115] Gelonin (GAP31) was covalently coupled to methoxypoly (ethylene glycol) (mPEG) 2000, mPEG 5000 and mPEG succinimidyl succinate 20K (SS-20PEG). mPEG does not affect the positive charge of protein, while charges alteration may result in lowering biological activity.…”