The Early Drugability Study Of Two Indolocarbazoles From Ocean On Anticancer Effects

Protein kinase C （PKC） is a class of structurally similar and functionally distinctserine/threonine kinases in animal cells, which has a central role in cellular signalingtransduction involved in cell proliferation,differentiation, apoptosis and angiogenesisas a signaling molecule. Consequently, PKC has become a therapeutic target for thetreatment of cancer.Different strategies have been devised in the drug development of PKCinhibitors and several PKC inhibitors are in the clinical trial now, such as UCN-01,PKC412, rottlerin and Bryostatin-1. However, there are still many problems.Improvement of isozyme specificity is the key point in PKC inhibitor research.MDZ-03is an indolecarbazole compound,which performs a good inhibitioneffect on PKCβⅡphosphorylation and also shows significant anticancer effect in vivoin the previous works in our lab. ZWM127is a Bisindolylmaleimide,which showsgood activity in screening model for PKCβII inhibitors and good solubility. Thispaper aims at the further study of anticancer effects of MDZ-03and ZWM127,as wellas the early druggability of MDZ-03.2. The early drugability study of MDZ-03on anticancer effectsWe mainly study the early drugability of PKC inhibitor MDZ-03, including itsantitumor effort in vivo,early safety evaluation and pharmacokinetic study.（1）In H22liver cancer model and A549human lung adenocarcinoma xenograftmodel, intraperitoneal injection twice a day, MDZ-03showed better anticancer effect.However, MDZ-03still has the problem of poor solubility, mouse of high-dose（20mg/kg） group died in the administration of the late, obvious weight loss.（2）The maximum dosage of MDZ-03was120mg/kg. The micronucleus testshowed MDZ-03didn’t induce genetic damage compared with the solvent group.（3）Establish the analytical method detecting MDZ-03of plasma biologicalsamples by high performance liquid chromatography （HPLC）, The chromatographicsystem provides good separation of MDZ-03with less interference. Draw the tumor-bearing mice （H22） a single dose of intraperitoneal injection of30mg/kgMDZ-03pharmacokinetic curves, calculation of pharmacokinetic parameters. Themain pharmacokinetic parameters of MDZ-03: t1/2β（min）:147.3;Tpeak（min）:39.5;Cmax(mg·L^-1):2.39;CL(L·kg^-1·min^-1):0.067;AUC(mg·min·L^-1):456.66.（4）After MDZ-03incubated with the mouse liver microsome, we detected ametabolic product of MDZ-03at9.64min of retention time.（5）The CaCo-2cell monolayer model was established to investigate the uptakepotential of MDZ-03, the results showed that MDZ-03intake less in this model anddid not detected by HPLC, MDZ-03is not suitable for oral administration by initialjudgment.2. Study of ZWM127on anticancer effects in vivo and in vitroWe evaluate the antitumor effect of ZWM127and its mechanisms of action，wealso further evaluate the antitumor effect of ZWM127in vivo.（1）We evaluated the inhibition effect in a variety of cell lines using MTT method,the results reavealed that ZWM127showed selective inhibition on cancer celllines,especially K562and A549cell lines.（2）FCM analysis revealed that ZWM127may arrest K562cells and A549cells atG2-M phase specifically.（3）ZWM127induced apoptosis via activating the mitochondrial pathway inK562and A549cells，its mechanism was involved in down-regulation of PKCβII and PKCθas well as PKC downstream p-AKT,p-GSK-3β and p-ERK1/2.（4）ZWM127has a certain degree of inhibition on PKC subtypes at the molecularlevel in vitro, among which PKCθ was the most significant.（5）ZWM127inhibited the NF-kb pathway.（6）ZWM127did not show a significant anticancer effect in H22solid tumor andnude mice PbaBe-PKCβII-Hela transplanted tumors in vivo, because ZWM127cannot exist stably in the plasma which converted into no active product in the bodyrapidly.