Influence of the pendant groups on electrochemical water oxidation catalyzed by cobalt(II) triazolylpyridine complexes.

The development of low-cost catalysts for the water oxidation reaction (WOR) is important for solving the bottleneck issues in water splitting and benefits the widespread utilization of renewable energy sources. Herein, four cobalt(II) triazolylpyridine complexes, namely [Co(DTE)2(H2O)2](ClO4)2·CH3COCH3 (1), [Co(DTE)2Cl2]·2CH3OH (2) (DTE = (1-(2-acetoxymethyl)-4-(2-pyridyl)1,2,3-triazole), [Co(DTEL)2(CH3OH)2](ClO4)2 (3), and [Co(DTEL)2Cl2]·H2O (4) (DTEL = (1-(2-hydroxy)-4-(2-pyridyl)1,2,3-triazole), were synthesized and characterized. The crystal structures of 1-3 were determined by X-ray single crystal diffraction analysis. The electrocatalytic water oxidation by 1-4 was studied in 0.1 M NaOAc-HOAc solutions. Complexes 1-4 were single-site molecular catalysts for the WOR under near-neutral conditions. The overpotentials for the WOR were 440 mV and 480 mV. The faradaic efficiencies were 77-92%. The rate constants kcat were 0.21-0.96 s-1. The catalytic activities were affected by the pendant groups of DTE and DTEL. Complexes with DTE (1 and 2) showed better activities than those with DTEL (3 and 4). Moreover, complexes 1-4 adsorbed on indium-doped tin oxide (ITO) and glassy carbon electrode surfaces were active for the WOR. A mechanism was proposed for the WOR catalyzed by 1-4.

Effect of carbamylation on the molecular recognition action of amino benzothiazole by carrier protein.

Increasing benzothiazole derivatives containing amino or N-acyl structures in position 2 have been largely developed as pesticides and medicines. However, the structure-function relationship of 2-substituted benzothiazole derivatives has seldom been illustrated from the perspective of their albumin-binding nature. Herein, to probe the influence of carbamylation on the albumin-binding nature of benzothiazole derivatives, formyl group was introduced to the amine group of 2-amino benzothiazole (ABT) to yield a novel modified ABT (MABT). Their protein-binding properties were systematically deciphered by spectroscopy, molecular modeling and density functional theory (DFT) calculations. The interaction mechanisms, recognition thermodynamics and binding geometry were investigated and compared. The structural alteration of human serum albumin was explored using synchronous fluorescence emission and circular dichroism spectrum technologies. Based on experimental results, the structures of protein complex with MABT and ABT were revealed by molecular docking method. The differences in energy transfer efficiency and molecular orientation of ABT and MABT in new complexes were tentatively explained by DFT calculations. The work was expected to help to understand the impact of different substituents on the bioactivity of benzothiazole derivatives and guide for structural designs of new compounds.

Novel Quinoline-based Ir(III) Complexes Exhibit High Antitumor Activity in Vitro and in Vivo.

Eight novel Ir(III) complexes listed as [Ir(H-P)2(P)]PF6 (PyP-Ir), [Ir(H-P)2(dMP)]PF6 (PydMP-Ir), [Ir(H-P)2(MP)]PF6 (PyMP-Ir), [Ir(H-P)2(tMP)]PF6 (PytMP-Ir), [Ir(MPy)2(P)]PF6 (MPyP-Ir), [Ir(MPy)2(dMP)]PF6 (MPydMP-Ir), [Ir(MPy)2(MP)]PF6 (MPyMP-Ir), [Ir(MPy)2((tMP)]PF6 (MPytMP-Ir) with 2-phenylpyri-dine (H-P) and 3-methyl-2-phenylpyridine (MPy) as ancillary ligands and pyrido-[3,2-a]-pyrido[1',2':1,2]imidazo[4,5-c]phenazine (P), 12,13-dimethyl pyrido-[3,2-a]-pyrido[1',2':1,2]-imidazo-[4,5-c]-phenazine (dMP), 2-methylpyrido [3,2-a]-pyrido-[1',2':1,2]-imidazo-[4,5-c]-phenazine (MP), and 2,12,13-trimethylpyrido-[3,2-a]-pyrido-[1',2':1,2]-imidazo-[4,5-c]-phenazine (tMP) as main ligands, respectively, were designed and synthesized to fully characterize and explore the effect of their toxicity on cancer cells. Cytotoxic mechanism studies demonstrated that the eight Ir(III) complexes exhibited highly potent antitumor activity selectively against cancer cell lines NCI-H460, T-24, and HeLa, and no activity against HL-7702, a noncancerous cell line. Among the eight Ir(III) complexes, MPytMP-Ir exhibited the highest cytotoxicity with an IC50 = 5.05 ± 0.22 nM against NCI-H460 cells. The antitumor activity of MPytMP-Ir in vitro could be contributed to the steric or electronic effect of the methyl groups, which induced telomerase inhibition and damaged mitochondria in NCI-H460 cells. More importantly, MPytMP-Ir displayed a superior inhibitory effect on NCI-H460 xenograft in vivo than cisplatin. Our work demonstrates that MPytMP-Ir could potentially be developed as a novel potent Ir-based antitumor drug.

Spectroscopic and computational exploration of hypoxanthine riboside interacting with plasma albumin.

Hypoxanthine riboside (HXR) is a nucleoside essential for wobble base pairs to translate the genetic code. In this work, an absorption and luminescence study showed that HXR and human serum albumin (HSA) formed a new complex through hydrogen bonds and van der Waals forces at ground state. Fluorescence probe experiments indicated that HXR entered the first subdomain of domain II in HSA and was fixed by amino acid residues in site I defined by Sudlow, and after competing with a known site marker. The recognition interaction featured negative ΔHÏ´ , ΔSÏ´ and ΔGÏ´ thermodynamic parameters. Fluorescence and circular dichroism spectra described the polarity of residues and α-helix and ß-strand content changed because of HXR binding. The most rational structure for the HXR-HSA complex was recommended by the molecular docking method, in which the binding location, molecular orientation, adjacent amino acid residues, and hydrogen bonds were included. In addition, the influence of ß-cyclodextrin and some essential metal ions on the balance of the HSA-HXR system interaction was measured. The study gained comprehensive information on the transportation mechanism for HXR in blood, and was of great significance in understanding the theory of HXR biotransformation and in discussing its clinical in vivo half-life.

Study on the molecular recognition action of lamivudine by human serum albumin.

In this work, the interaction of an anti-HIV drug lamivudine and human serum albumin (HSA) was studied by multispectroscopic and molecular modeling methods. The fluorescence emission spectra showed that the fluorescence of HSA was quenched by lamivudine through static mechanism with HSA-lamivudine complex produced at ground state. According to the binding equilibriums observed at 4 different temperatures, the number of binding site, binding constant, enthalpy change, entropy change, and Gibbs free energy change of the interaction were calculated. The results indicated that there was only 1 main binding site under present concentration condition, and then, the location of this binding site was ascertained by molecular probe experiments using warfarin and ibuprofen as site markers. The interaction was a spontaneous and exothermic process. Hydrogen bonds and van der Waals force were the major power that fixed lamivudine on Sudlow's site I in subdomain IIA of HSA molecule. The distance between donor and acceptor was determined by Förster's nonradiative fluorescence resonance energy transfer theory. Circular dichroism spectra exhibited the alteration of HSA's secondary structures. Molecular modeling investigation revealed the structure of HSA-lamivudine complex, including the conformation of lamivudine in binding site, the amino residues close to lamivudine, and the interaction forces between receptor and ligand. The study may be beneficial to therapists in understanding the distribution of lamivudine in vivo and explaining its drug-resistant mechanism in clinical diagnosis.

Comparison and analysis on the serum-binding characteristics of aspirin-zinc complex and aspirin.

This study was designed to compare the protein-binding characteristics of aspirin-zinc complex (AZN) with those of aspirin itself. AZN was synthesized and interacted with a model transport protein, human serum albumin (HSA). Three-dimensional fluorescence, ultraviolet-visible and circular dichroism (CD) spectra were used to characterize the interaction of AZN with HSA under physiological conditions. The interaction mechanism was explored using a fluorescence quenching method and thermodynamic calculation. The binding site and binding locality of AZN on HSA were demonstrated using a fluorescence probe technique and Förster non-radiation energy transfer theory. Synchronous fluorescence and CD spectra were employed to reveal the effect of AZN on the native conformation of the protein. The HSA-binding results for AZN were compared with those for aspirin under consistent experimental conditions, and indicated that aspirin acts as a guide in AZN when binding to Sudlow's site I, in subdomain IIA of the HSA molecule. Moreover, compared with aspirin, AZN showed greater observed binding constants with, but smaller changes in the α-helicity of, HSA, which proved that AZN might be easier to transport and have less toxicity in vivo.

High prevalence of aspirin resistance in elderly patients with cardiovascular disease and metabolic syndrome.

BACKGROUND: Metabolic syndrome is known to be a prothrombotic state. We undertook this study to examine a hypothesis that aspirin resistance may be associated with metabolic syndrome, and to assess other potential determinants of aspirin resistance in patients with cardiovascular disease (CVD). METHODS: A total of 469 elderly patients with CVD were recruited. One hundred and seventy-two patients with metabolic syndrome and 297 without metabolic syndrome (control group) received daily aspirin therapy (≥ 75 mg) over one month. Platelet aggregation was measured by light transmission aggregometry (LTA). Aspirin resistance was defined as ≥ 20% arachidonic acid (AA)- and ≥ 70% adenosine diphosphate (ADP)-induced aggregation according to LTA. Aspirin semi-responders were defined as meeting one (but not both) of these criteria. RESULTS: By LTA, 38 of 469 (8.1%) patients were aspirin resistant. The prevalence of aspirin resistance was higher in the metabolic syndrome group compared with the control group [11.6 % vs. 6.6%, odds ratio (OR) = 2.039; 95% confidence interval (CI): 1.047-3.973]. In the multivariate logistic regression analysis, metabolic syndrome (OR = 4.951, 95% CI: 1.440-17.019, P = 0.011) was a significant risk factor for aspirin resistance. CONCLUSIONS: A significant number of patients with CVD and metabolic syndrome are resistant to aspirin therapy. This might further increase the risk of cardiovascular morbidity and mortality in these patients.

Investigation on the protein-binding properties of icotinib by spectroscopic and molecular modeling method.

Icotinib is a highly-selective epidermal growth factor receptor tyrosine kinase inhibitor with preclinical and clinical activity in non-small cell lung cancer, which has been developed as a new targeted anti-tumor drug in China. In this work, the interaction of icotinib and human serum albumin (HSA) were studied by three-dimensional fluorescence spectra, ultraviolet spectra, circular dichroism (CD) spectra, molecular probe and molecular modeling methods. The results showed that icotinib binds to Sudlow's site I in subdomain IIA of HSA molecule, resulting in icotinib-HSA complexes formed at ground state. The number of binding sites, equilibrium constants, and thermodynamic parameters of the reaction were calculated at different temperatures. The negative enthalpy change (ΔH(θ)) and entropy change (ΔS(θ)) indicated that the structure of new complexes was stabilized by hydrogen bonds and van der Waals power. The distance between donor and acceptor was calculated according to Förster's non-radiation resonance energy transfer theory. The structural changes of HSA caused by icotinib binding were detected by synchronous spectra and circular dichroism (CD) spectra. Molecular modeling method was employed to unfold full details of the interaction at molecular level, most of which could be supported by experimental results. The study analyzed the probability that serum albumins act as carriers for this new anticarcinogen and provided fundamental information on the process of delivering icotinib to its target tissues, which might be helpful in understanding the mechanism of icotinib in cancer therapy.

Protein-binding properties of a designed steroidal lactam compound.

Introducing amide bonds into a steroid nucleus or its side chain may reduce the acute toxicity and enhance the pharmaceutical activity. In this work, a designed steroidal amide compound, named 3ß-hydroxy-17-aza-d-homo-5-androsten-17-one (HAAO), was synthesized and identified. The interactions between HAAO and human serum albumin (HSA) were studied by multiple spectroscopic methods and molecular modeling procedures. It was found that HAAO locates in Sudlow's site I in subdomain IIA of HSA molecules, relying on hydrogen bonds and van der Waals power to form HAAO-HSA complexes at ground state. The number of binding sites, binding constants, enthalpy change (ΔH(θ)), Gibbs free energy change (ΔG(θ)) and entropy change (ΔS(θ)) were calculated at different temperatures based on fluorescence quenching theory and classical thermodynamic equation. The percentages content of the HSA's secondary structures in presence of HAAO were detected by circular dichroism (CD) spectra and compared with those in no presence of HAAO. In addition, the experimental results of both binding site and conformational change were further confirmed by molecular modeling investigation, in which more details of the binding were visually unfolded. The information provided by the study may be useful for designing novel chemotherapeutic drugs and be helpful both in the early stages of drug discovery and in clinical practice.

Synthesis and properties of the cyano complex of oxo-centered triruthenium core [Ru3(µ3-O)(µ-CH3COO)6(pyridine)2(CN)].

The preparation and properties of a new cyano complex containing the Ru3(µ3-O) core, [Ru3(µ3-O)(µ-CH3COO)6(py)2(CN)] (1; py = pyridine), are reported. Complex 1 in CH2Cl2 showed intense absorption bands at 244, 334, and 662 nm, corresponding to a π-π* transition of the ligand, cluster-to-ligand charge transfer, and intracluster transitions, respectively. The cyclic voltammogram of 1 in 0.1 M (n-Bu)4NPF6-CH2Cl2 showed redox waves for the processes Ru3(II,II,III)/Ru3(II,III,III), Ru3(II,III,III)/Ru3(III,III,III), and Ru3(III,III,III)/Ru3(III,III,IV) at E1/2 = -1.49, -0.26, and +1.03 V vs Ag/AgCl, respectively. The first two redox potentials are more negative by ca. 0.2 V in comparison with the corresponding potentials of [Ru3(µ3-O)(µ-CH3COO)6(py)3](+). This is in sharp contrast to the positive shifts of the corresponding waves of [Ru3(II,III,III)(µ3-O)(µ-CH3COO)6(py)2(CO)]. Density functional theory (DFT) calculations of [Ru3(II,III,III)(µ3-O)(µ-CH3COO)6(py)3], [Ru3(II,III,III)(µ3-O)(µ-CH3COO)6(py)2(CN)](-), and [Ru3(II,III,III)(µ3-O)(µ-CH3COO)6(py)2(CO)] showed that the positive charge of the ruthenium is delocalized over the triruthenium cores of the first two and is localized as Ru(II)(CO){Ru(III)(py)}2 in the CO complex. The calculations explain the difference in the π interactions of the two ligands with the triruthenium cores.

Proton-coupled electron transfer and Lewis acid recognition at self-assembled monolayers of an oxo-bridged diruthenium(III) complex functionalized with two disulfide anchors.

A new µ-oxo-bis(µ-acetato)diruthenium(III) complex bearing two pyridyl disulfide ligands {[Ru2(µ-O)(µ-OAc)2(bpy)2(L(py-SS))2](PF6)2 (OAc = CH3CO2(-), bpy = 2,2'-bipyridine, L(py-SS) = (C5H4N)CH2NHC(O)(CH2)4CH(CH2)2SS) (1)} has been synthesized to prepare self-assembled monolayers (SAMs) on the Au(111) electrode surface. The SAMs have been characterized by contact-angle measurements, reflection-absorption surface infrared spectroscopy, cyclic voltammetry, and reductive desorption experiments. The SAMs exhibited proton-coupled electron transfer (PCET) reactions when the electrochemistry was studied in aqueous electrolyte solution (0.1 M NaClO4 with Britton-Robinson buffer to adjust the solution pH). The potential-pH plot (Pourbaix diagram) in the pH range from 1 to 12 has established that the dinuclear ruthenium moiety was involved in the interfacial PCET processes with four distinct redox states: Ru(III)Ru(III)(µ-O), Ru(II)Ru(III)(µ-OH), Ru(II)Ru(II)(µ-OH), and Ru(II)Ru(II)(µ-OH2). We also demonstrated that the interfacial redox processes were modulated by the addition of Lewis acids such as BF3 or Al(3+) to the electrolyte media, in which the externally added Lewis acids interacted with µ-O of the dinuclear moiety within the SAMs.

Terpyridine platinum(II) complexes containing triazine di- or tri-thiolate bridges: structures, luminescence, electrochemistry, and aggregation.

Dinuclear complexes [{Pt(trpy)}(2)(L)](PF(6))(2) (trpy = 2,2':6',2''-terpyridine, L = 2-octylthio-1,3,5-triazine-4,6-dithiolate ion (1), L = 2-octadecylthio-1,3,5-triazine-4,6-dithiolate ion (2), L = 2-di-n-butylamino-1,3,5-triazine-4,6-dithiolate ion (3)) and a trinuclear complex [{Pt(trpy)}(3)(L)](PF(6))(3) (L = 1,3,5-triazine-2,4,6-trithiolate ion (4)) have been synthesized and characterized. The single crystal X-ray analysis revealed that the two {Pt(trpy)}(2+) fragments in 1 and 3 adopt a syn-configuration. The PtPt distances are around 4.3 Å, suggesting no intramolecular PtPt interactions. Complexes 1-4 in acetonitrile show broad absorption bands at around 470 nm, assigned to mainly the ligand-to-ligand charge transfer ((1)LLCT) from triazine thiolates to trpy based on the comparison to the related complexes and the density functional theory (DFT) calculations. The red luminescence of 1-4 in acetonitrile is attributable to emission predominantly from (3)LLCT. Cyclic voltammograms of 1-3 exhibit four redox couples from -2.0 V to 0 V vs. Ag/AgCl. The two consecutive processes at around -0.70 V are assigned to the sequential reduction of two trpy ligands. This assignment was further supported by the observation of the anion radical of trpy in spectroelectrochemical experiments. The splitting of the redox potentials of two trpy ligands evidences the moderate electronic coupling interactions mediated by the triazine dithiolate bridges. Complex 2 formed a transparent red gel in CH(3)CN, whereas 4 produced a gel-like solid in the mixtures of CH(3)CN and other solvents. The interactions dominating the aggregative behaviours have been discussed based on the results of electronic absorption and emission spectroscopy.

Biophysical influence of isocarbophos on bovine serum albumin: spectroscopic probing.

Isocarbophos (ICP) is a phosphorous pesticide with high toxicity. It has been detected in several kinds of food and therefore can enter human body. In this paper, spectroscopic approaches including three-dimensional fluorescence (3D-FL) spectroscopy, UV-visible absorption spectroscopy and circular dichroism (CD) spectroscopy were employed to explore the binding of ICP to bovine serum albumin (BSA) at simulated physiological conditions. It was found that the fluorescence quenching of BSA was caused by the formation of ICP-BSA complex at ground state and belonged to static quenching mechanism. The binding constants, the number of binding sites, enthalpy change (ΔH(θ)), Gibbs free energy change (ΔG(θ)) and entropy change (ΔS(θ)) were calculated at four different temperatures according to Scatchard model and thermodynamic equations. To identify the binding location, fluorescence probe techniques were used. The results showed that warfarin, an acknowledged site marker for BSA, could be partially replaced by ICP when ICP was added to warfarin-BSA systems, which demonstrated that ICP primarily bound on Sudlow's site I in domain IIA of BSA molecule. The distance r (3.06 nm) between donor (Trp-212) and acceptor (ICP) was obtained based on Förster's non-radiation fluorescence resonance energy transfer (FRET) theory. Furthermore, the CD spectral results indicated that the secondary structure of BSA was changed in presence of ICP. The study is helpful to evaluating the toxicology of ICP and understanding its effects on the function of protein during the blood transportation process.

Biochemical evaluation of a synthesized isoflavone-selenium complex by molecular spectra.

A coordination compound of 5, 7-dihydrox-4'-methoxyisoflavone and selenium was synthesized and its structure was identified by IR, LC-MS and (1)H-NMR. Its biochemical effects were investigated using bovine serum albumin (BSA) as a target protein molecule, in which process three-dimensional (3D) fluorescence spectra, ultraviolet spectra, circular dichroism (CD) spectra and fluorescence probe techniques were employed. The interaction of SEIF and BSA was discussed by fluorescence quenching method and Förster non-radiation energy transfer theory. The thermodynamic parameters ΔH (θ), ΔG (θ), ΔS (θ) at different temperatures were calculated according to Van't Hoff isobaric equation and the results indicated the interaction was an exothermic as well as a spontaneous process. The binding site was explored by fluorescence probe method using warfarin and ibuprofen as markers. Intramolecular forces which are responsible for maintaining the binding were mainly hydrogen bond and van der Waals power. The average distance from the tryptophan residue in domain II of BSA (donor) to SEIF (acceptor) is 3.57 nm at body temperature. The conformation changes of BSA were investigated by 3D fluorescence and CD spectra.

Chemicobiological effects of herbicide MCPA-Na on plasma proteins.

Under physiological conditions, the potential hematological toxicity of herbicide 2-methyl-4-chlorophenoxyacetic acid sodium (MCPA-Na) was discussed by fluorescence probe technology and spectroscopy methods including three-dimensional (3D) fluorescence, UV absorption and circular dichroism (CD) spectra. In vitro, MCPA-Na bound with bovine serum albumin (BSA) and formed new complex at ground state by electrostatic force and hydrogen bond. During the process, non-radiation energy transfer from BSA to MCPA-Na occurred and the distance r between donor and acceptor was obtained based on Förster theory. The binding site was investigated by fluorescence probe method and the results implied MCPA-Na was absorbed on domain II of BSA molecule. The enthalpy change (ΔH(θ)), Gibbs free energy change (ΔG(θ)) and entropy change (ΔS(θ)) were calculated at four different temperatures according to Van't Hoff isobar equation and Gibbs-Helmholtz equation. Negative value of ΔG(θ) indicated the process of binding was a spontaneous and irreversible process, which gave a broad hint that MCPA-Na was likely to be poisonous. CD spectra exhibited significant changes of secondary structures in BSA molecule and three-dimensional fluorescence spectra indicated the tryptophan residue in BSA was placed in a less hydrophobic environment, which presented additional evidence to caution the danger of MCPA-Na residue in food. Meanwhile, the mechanism and geometry of the binding was analyzed at molecular level.

Synthesis of vitamin-selenium complex and its effects on proteins and tumor cells.

A selenium-vitamin P complex (SEVP) was synthesized and its structure was determined by IR, LC-MS and (1)H NMR. Its biological effects on bovine serum albumin (BSA) and human colon carcinoma tumor cells were studied by molecular spectra, MTT and flow cytometry. The interaction of SEVP and BSA was discussed by fluorescence quenching method and Förster non-radiation energy transfer theory. The thermodynamic parameters ΔH(θ), ΔG(θ), ΔS(θ) at different temperatures were calculated and the results indicate the interaction is an exothermic as well as entropy-driven process. Hydrogen bond and electrostatic force played major role in the reaction. The binding geometry and conformation changes of BSA were investigated by fluorescence probe technique and circular dichroism (CD) spectra. The effects of SEVP on human colon carcinoma tumor cells HT29 were tested by MTT method and flow cytometry (FCM). The MTT results show the proliferation of HT29 tumor cells were inhibited by SEVP and the inhibition was associated with dose and time. The FCM analysis disclosed SEVP interrupted the DNA synthesis of tumor cells at S phase.

A butadiyne-linked diruthenium molecular wire self-assembled on a gold electrode surface.

A 1,3-butadiyne-linked diruthenium complex 4 is successfully brought onto the gold surface in a lying flat mode to form self-assembled monolayers (SAMs) showing reversible multiple redox behaviors on the electrode surface. The diruthenium species with different oxidation states, particularly the Ru(2)(III,III) state which is unstable and impossible to isolate from the solution, can be detected by in situ IR spectroscopy.

Synthesis of morin-zinc(II) complex and its interaction with serum albumin.

A morin-zinc(II) complex (MZ) was synthesized and its interaction with bovine serum albumin (BSA) were studied by molecular spectroscopy including fluorescence emission spectra, UV-visible spectra, circular dichroism (CD) spectra, three-dimensional fluorescence spectra, and synchronous fluorescence spectra. The interaction mechanism of BSA and MZ was discussed by fluorescence quenching method and Förster non-radiation energy transfer theory. The thermodynamic parameters ΔHθ, ΔGθ, ΔSθ at different temperatures were calculated and the results indicate the interaction is an exothermic as well as entropy-driven process. Hydrogen bond forces played the most important role in the reaction. The fluorescence probe experiment showed that the binding site of MZ is in subdomain IIA of BSA and the distance between BSA and MZ is 3.17 nm at normal body temperature. The conformation changes of BSA in presence of MZ were investigated by CD spectra and three-dimensional fluorescence spectra.

Prepubertal genistein exposure affects erbB2/Akt signal and reduces rat mammary tumorigenesis.

Breast cancer is the most common malignancy in women worldwide and pharmaceutical agents have therapeutic and preventive effects in breast cancer. The human epidermal growth factor receptor 2/neu is one of the most important oncogenes in human breast cancer. Prepubertal exposure to endogenous estradiol and a phytoestrogen, genistein (Gen), has been shown to reduce future breast cancer risk. Gen downregulates tyrosine kinase regulated protein expression and reduces prostate cancer. In this study, the effects of prepubertal exposure to Gen on rat mammary carcinogenesis and the erbB2/Akt signal pathway were investigated. Prepubertal female Sprague-Dawley rats were daily exposed to Gen at 125 mg/kg (Gen-1) and 500 mg/kg (Gen-5) from postnatal days 22-28. Subsequently, the rats were given a single dose of 100 mg/kg 7.12-dimethylbenz [a] anthracene on postnatal day 42 to induce mammary tumor. The mRNA expression of erbB2 and amplified in breast cancer 1 (AIB1) was detected by reverse transcription-polymerase chain reaction. The protein levels of proliferating cell nuclear antigen (PCNA), erbB2, phosphotyrosine protein, Akt, and p-Akt were detected by immunohistochemistry and Western blotting. The activity of protein tyrosine kinase (PTK) was detected by liquid scintillation counting. The percentage of rats with mammary tumors in breast cancer model (BCM), Gen-1, and Gen-5 was 71.43, 52.38, and 33.34%, respectively. The incidence of 7.12-dimethylbenz [a] anthracene-induced mammary tumors significantly decreased in Gen-5 compared with that in BCM. The mRNA levels of AIB1 and erbB2 and the protein levels of erbB2, p-Akt, and PCNA protein expression were downregulated for a long time in the mammary tumors in Gen-5 groups. The activity of PTK was also decreased for a long time. However, the total Akt protein expression did not change significantly among BCM, Gen-1, and Gen-5. Prepubertal exposure of Sprague-Dawley female rats to 500 mg/kg Gen can reduce later breast cancer risk and its protective effect is associated with persistent downregulation of the expression of erbB2, p-Akt, AIB1, and PCNA and with low PTK activity in the mammary tumor. Our results suggest that erbB2/Akt signaling plays a role in tumor formation and targeting erbB2/Akt signaling with prepubertal exposure to Gen may provide greater efficacy to the current therapies used to treat tumors.