A review on medicinal importance,pharmacological activity and bioanalytical aspects of beta-carboline alkaloid "Harmine"

Harmine,a beta-carboline alkaloid,is widely distributed in the plants,marine creatures,insects, mammalians as well as in human tissues and body fluids.Harmine was originally isolated from seeds of Peganum harmal in 1847 having a core indole structure and a pyridine ring.Harmine has various types of pharmacological activities such as antimicrobial,antifungal,antitumor,cytotoxic, antiplasmodial,antioxidaant,antimutagenic,antigenotoxic and hallucinogenic properties.It acts on gamma-aminobutyric acid type A and monoamine oxidase A or B receptor,enhances insulin sensitivity and also produces vasorelaxant effect.Harmine prevents bone loss by suppressing osteoclastogenesis.The current review gives an overview on pharmacological activity and analytical techniques of harmine,which may be useful for researcheres to explore the hidden potential of harmine and and will also help in developing new drugs for the treatment of various diseases.

Harmine induces apoptosis in HepG2 cells via mitochondrial signaling pathway

BACKGROUND:Harmine has antitumor and antinociceptive effects,and inhibits human DNA topoisomerase.However no detailed data are available on the mechanisms of action of harmine in hepatocellular carcinoma.This study aimed to investigate the effects of harmine on proliferation and apoptosis and the underlying mechanisms in the human hepatocellular carcinoma cell line HepG2.METHODS:The proliferation of HepG2 cells was determined by the cell counting kit-8 (CCK-8) assay and the clone formation test.The morphology of HepG2 cells was examined using fluorescence microscopy after Hoechst 33258 staining Annexin V/propidium iodide (PI) was used to analyze apoptosis and PI to analyze the cell cycle.Western blotting was used to assess expression of the apoptosis-regulated genes Bcl-2,Bax,Bcl-xl,Mcl-1,caspase-3,and caspase-9 Mitochondrial transmembrane potential (Ψ m) was determined using JC-1.RESULTS:Harmine inhibited the proliferation of HepG2 cells in a dose-dependent manner.Hoechst 33258 staining revealed nuclear fragmentation and chromosomal condensation,cell shrinkage,and attachment loss in HepG2 cells treated with harmine.The percentage of the sub/G1 fraction was increased in a concentration-dependent manner,indicating apoptotic cell death.PI staining showed that harmine changed the cell cycle distribution,by decreasing the proportion of cells inG0/G1 and increasing the proportion in S and G2/M.Harmine induced apoptosis in a concentration-dependent manner,with rates of 20.0%,32.7% and 64.9%,respectively.JC-1 revealed a decrease in Ψ m.Apoptosis of HepG2 cells was associated with caspase-3 and caspase-9 activation,down-regulation of Bcl-2,Mcl-1,and Bcl-xl,and no change in Bax.CONCLUSIONS:Harmine had an anti-proliferative effect in HepG2 cells by inducing apoptosis.Mitochondrial signal pathways were involved in the apoptosis.The cancer-specific selectivity shown in this study suggested that harmine is a promising novel drug for human hepatocellular carcinoma.

Heme perbxidases are responsible for the dehydrogenation and oxidation metabolism of harmaline into harmine

Harmaline and harmine areβ-carboline alkaloids with effective pharmacological effects.Harmaline can be transformed into harmine after oral administration.However,enzymes involved in the metabolic pathway remain unclear.In this study,harmaline was incubated with rat liver microsomes(RLM),rat brain microsomes(RBM),blood,plasma,broken blood cells,and heme peroxidases including horseradish peroxidase(HRP),lactoperoxidase(LPO),and myeloperoxidase(MPO).The production of harmine was determined by a validated UPLC-ESI-MS/MS method.Results showed that heme peroxidases catalyzed the oxidative dehydrogenation of harmaline.All the reactions were in accordance with the Hill equation.The reaction was inhibited by ascorbic acid and excess H_(2)0_(2).The transformation of harmaline to harmine was confirmed after incubation with blood,plasma,and broken blood cells,rather than RLM and RBM.Harmaline was incubated with blood,plasma,and broken cells liquid for 3 h,and the formation of harmine became stable.Results indicated an integrated metabolic pathway of harmaline,which will lay foundation for the oxidation reaction of dihydro-P-carboline.Moreover,the metabolic stability of harmaline in blood should not be ignored when the pharmacokinetics study of harmaline is carried out.

Preparation, characterization and evaluation of liposomal doxorubicin and harmine that show synergistic activity in vitro

A combinatory therapeutic system that simultaneously targets several independent pathways is preferred for the treatment of cancer. In our study, a combinatory liposomal delivery system containing doxorubicin （Dox） and harmine （HM） was constructed by thin film dispersing method together with pH gradient method. A simple, precise and accurate spectrophotometric method for the determination of Dox and HM in liposomal formulation was established and validated. A drug HSPC ratio of 1： 20, loading time of 30 min and loading temperature of 50 ~C were the optimal conditions for the preparation of drug loaded liposomes, which exhibited excellent physicochemical properties such as average particle size of-100 nm, low polydispersity index below 0.2 and high entrapment efficiency above 93%. Sustained release of drug from liposomes at pH 7.4 showed good biological safety. The synergetic cytotoxic effect for these two drugs was evaluated in MCF-7 ceils. The in vitro antitumor studies demonstrated the superior anti-proliferation activity of the liposomal Dox and HM with a combination index of 0.81, which indicated great synergistic effect and increased anti-proliferation efficiency. The experimental data suggested that combinational liposome therapy could be an effective way to develop efficient treatment of cancers.

Effects of 1-(2-chlorine)-phenyl-9-butyl-β-carboline(DH-330)on Cystic Echinococcosis in Mice in vivo

[Objectives]The paper was to evaluate the in vivo treatment efficacy of harmine derivative 1-(2-chloro)phenyl-9-butyl-β-carboline(DH-330)on mice cystic echinococcosis(CE).[Methods]Kunming mice were injected intraperitoneally into the protoscoleces and infected with secondary infection for 8 months to prepare CE model.The successfully modeled mice were randomly divided into 6 groups according to their body weight:model group,control-1,-2 groups and experimental-L,-M,-H groups,with 10 mice in each group.The model group was given distilled water and control-1,-2 groups were given 50 mg/kg albendazole and harmine,respectively.The experimental-L,-M,-H groups were given 25,50 and 100 mg/kg DH-330.After 8 weeks of intragastric administration,the mice were dissected and vesicles were taken,and the differences of cyst weight were compared.The ultrastructure changes of cysts were observed by transmission electron microscope(TEM).The histopathology of cysts and liver were observed by hematoxylin-eosin(HE)staining method.[Results]The cyst weight of model group,control-1,-2 groups and experimental-L,-M,-H groups were(34.38±4.32),(11.38±2.37),(15.89±1.31),(16.22±2.30),(11.69±2.95)and(9.78±1.14)g,respectively.Compared between drugs group and model group,the difference was significant(all P<0.05);compared between experimental-H group and harmine group,the difference was significant(P<0.05).Except for the model group and experimental-L group,all other groups can damage the hydatid nuclei,which lead to cell lysis and nucleoli disappear.Experimental groups can improve inflammatory cells infiltration in liver and vesicle.[Conclusions]DH-330 can reduce the cysts weight of CE mice,inhibit the growth of hydatid,and improve the inflammation of the liver and vesicles,showing a good resistance against Echinococcus granulosus,or may become an effective new drug against hydatid disease.

Synthesis and cytotoxic activity of 3-phenyl-4-substituted-β-carbolines

A new class of 3-phenyl-4-substituted-β-carbolines via iodine-mediated electrophilic cyclization as key step were synthesized and their inhibitory activity against three tumor cell lines was evaluated in vitro. It was found that some of the tested compounds showed better cytotoxicity against HeLa and MCF-7 cells than harmine.