V. P. Chekhonin

The neuroleptic activity of haloperidol increases after its solubilization in surfactant micelles: Micelles as microcontainers for drug targeting

It has been suggested to use surfactant micelles as microcontainers for increasing the efficiency of neuroleptic targeting from blood flow into the brain. The neuroleptic action of haloperidol, intraperitoneally injected into mice in micellar solution of non‐ionic block copolymer surfactant (pluronic P‐85) in water, increased several‐fold if compared with that observed for haloperidol aqueous solution. Incorporation of brain‐specific antibodies into haloperidol‐containing micelles resulted in additional drastic increase (more than by 2 orders of magnitude) in the drug effect.

A new class of drug carriers: micelles of poly(oxyethylene)-poly(oxypropylene) block copolymers as microcontainers for drug targeting from blood in brain☆

Abstract A new concept of design of drug delivery systems based on using self-assembling supramacromolecular complexes is formulated. Microcontainers for drug targeting were prepared using polymeric surfactant poly(oxyethylene)-poly(oxypropylene) block copolymer (pluronic). Molecules of a drug are solubilized in a pluronic micelle being incorporated into its inner hydrophobic core, formed by poly(oxypropylene) chain blocks. The outer hydrophilic shell of such micelles is formed by nontoxic and nonimmunogenic poly(oxyethylene) blocks. Solubilization of low molecular weight compounds (fluorescein isotbiocyanate (FITC), haloperidol etc.) in pluronic micelles was studied using fluorescence and ultracentrifugation. The dimensions of the aggregates formed in the solutions of various pluronics (P85, F64, L68, L101) and its mixtures were determined using quasielastic light-scattering technique. In a majority of cases the diameter of pluronic micelles (including those containing solubilized compounds) was in the range of 12–36 nm. For targeting of such microcontainers to a certain cell the pluronic molecules were conjugated with antibodies against a target-specific antigen or with protein ligands selectively interacting with target cell receptors. The obtained conjugates were then incorporated into the drug-containing micelles by simple mixing of the corresponding components. It was found that solubilization of FITC in pluronic micelles considerably influences its distribution in animal (mouse) tissues resulting, in particular, in the drastic increase of FITC fluorescence in lung. Conjugation of FITC-containing micelles with insulin vector results in increase of FITC penetration in all tissues including the brain. The specific targeting of the solubilized FITC in brain was observed in the case when the pluronic conjugate with antibodies to the antigen of brain glial cells ( α 2 -glycoprotein) was incorporated into micelles. Under these conditions the considerable increase of FITC fluorescence in the brain and decrease of its fluorescence in the lungs has been registered. Possibility of using micellar microcontainers for targeting of solubilized neuroleptics (haloperidol) in brain was studied. Incorporation of antibodies to α 2 glycoprotein into haloperidol-containing micelles results in a drastic increase of drug effect. This result indicates that vector-containing pluronic micelles provide an effective transport of solubilized neuroleptics across blood-brain barrier.

Genetic determinants of aggression and impulsivity in humans

Human aggression/impulsivity-related traits have a complex background that is greatly influenced by genetic and non-genetic factors. The relationship between aggression and anxiety is regulated by highly conserved brain regions including amygdala, which controls neural circuits triggering defensive, aggressive, or avoidant behavioral models. The dysfunction of neural circuits responsible for emotional control was shown to represent an etiological factor of violent behavior. In addition to the amygdala, these circuits also involve the anterior cingulated cortex and regions of the prefrontal cortex. Excessive reactivity in the amygdala coupled with inadequate prefrontal regulation serves to increase the likelihood of aggressive behavior. Developmental alterations in prefrontal-subcortical circuitry as well as neuromodulatory and hormonal abnormality appear to play a role. Imbalance in testosterone/serotonin and testosterone/cortisol ratios (e.g., increased testosterone levels and reduced cortisol levels) increases the propensity toward aggression because of reduced activation of the neural circuitry of impulse control and self-regulation. Serotonin facilitates prefrontal inhibition, and thus insufficient serotonergic activity can enhance aggression. Genetic predisposition to aggression appears to be deeply affected by the polymorphic genetic variants of the serotoninergic system that influences serotonin levels in the central and peripheral nervous system, biological effects of this hormone, and rate of serotonin production, synaptic release and degradation. Among these variants, functional polymorphisms in the monoamine oxidase A (MAOA) and serotonin transporter (5-HTT) may be of particular importance due to the relationship between these polymorphic variants and anatomical changes in the limbic system of aggressive people. Furthermore, functional variants of MAOA and 5-HTT are capable of mediating the influence of environmental factors on aggression-related traits. In this review, we consider genetic determinants of human aggression, with special emphasis on genes involved in serotonin and dopamine metabolism and function.

Contribution of microRNAs to radio- and chemoresistance of brain tumors and their therapeutic potential.

Glioblastomas, particularly high grade brain tumors such as glioblastoma multiforme, are characterized by increased anaplasy, malignancy, proliferation, and invasion. These tumors exhibit high resistance to radiation therapy and treatment with anti-cancer drugs. The radio- and chemoresistance of gliomas is attributed to cancer stem cells (CSCs) that are considered as major contributors for maintenance and propagation of tumor cell mass, cancer malignancy and invasiveness, and tumor cell survival after courses of radiotherapy and medical interventions. MicroRNAs (miRNAs), key post-transcriptional gene regulators, have altered expression profiles in gliomas. Some of miRNAs whose expression is markedly up-regulated in brain tumors are likely to have a pro-oncogenic role through supporting growth, proliferation, migration, and survival of cancer stem and non-stem cells. In contrast, a population of miRNA possessing anti-tumor effects is suppressed in gliomas. In this review, we will consider miRNAs and their influence on radio- and chemoresistance of gliomas. These miRNAs harbor a great therapeutic significance as potent agents in future targeted anti-cancer therapy to sensitize glioma tumor cells and CSCs to cytotoxic effects of radiation exposure and treatment with anti-cancer drugs.

VEGF in tumor progression and targeted therapy.

Progression of solid tumors depends on vascularization and angiogenesis in a malignant tissue. Among a whole range of proangiogenic factors, a vascular endothelial growth factor A (VEGF-A) plays a key role. Blockade of VEGF may lead to regression of vascular network and inhibition of a tumor growth. In the present time, bevacizumab has been introduced into wide clinical practice in therapy of breast cancer, colorectal cancer and recurrent high-grade gliomas (HGGs). Coadministration of antiangiogenic therapy with irinotecan may increase probability of the response to the treatment and prolong progression-free survival rate (PFS). Moreover, bevacizumab is well tolerated and significantly improves patients quality of life. However, in the case of brain tumors, the efficiency of such an approach is controversial. The antiangiogenic therapy can slightly delay tumor growth and does not lead to complete recovery. In addition, it contributes to enhanced tumor cell invasion into the normal brain. The mechanisms of resistance include activation of alternative proangiogenic signaling pathways, of an invasive population of tumor cells, metabolic change toward glycolysis and recruitment of myeloid bone marrow-derived cells to tumors. Obviously, that anti-VEGF therapy as monotherapy was not effective against HGGs. To enhance the antitumor treatment efficacy, it is necessary to develop a multi-target strategy to inhibit critical processes in malignancy progression such as angiogenesis, invasion, autophagy, metastatic spread, recruitment of bone marrow-derived endothelial cells and tumor stem-like cells. In addition, anti-VEGF antibodies have shown a promising result as a tumor-targeting vector for delivery therapeutic and diagnostic drugs in brain tumors.

Fatty acid acylated Fab-fragments of antibodies to neurospecific proteins as carriers for neuroleptic targeted delivery in brain

A method for targeted delivery of neuroleptics from blood in brain based on using Fab‐fragments of antibodies to antigens of brain glia cells (acid gliofibrillar antigen and α2‐glycoprotein) is suggested. The essence of the technique is that the molecule of neuroleptic (trifluoperazine) is conjugated with Fab‐fragments of these antibodies. The conjugate thus obtained is modified by stearoylchloride in the system of Aerosol OT reversed micelles in octane. The study of the distribution of 125I‐labelled conjugates in the rat organism after intracordial introduction is performed. On the contrary to the nonmodified conjugates and conjugate, containing fatty acylated Fab‐fragments of antibodies, nonspecific to the rat brain, the conjugate of trifluoperazine with stearoylated Fab‐fragments of antibodies to neurospecific antigens accumulate in brain tissues. The drastic increase of the neuroleptic activity of trifluoperazine resulting from its coupling with stearoylated Fab‐fragments of antiglial antibodies is observed.

VEGF-targeted magnetic nanoparticles for MRI visualization of brain tumor.

UNLABELLED This work is focused on synthesis and characterization of targeted magnetic nanoparticles as magnetic resonance imaging (МRI) agents for in vivo visualization of gliomas. Ferric oxide (Fe3O4) cores were synthesized by thermal decomposition and coated with bovine serum albumin (BSA) to form nanoparticles with Deff of 53±9nm. The BSA was further cross-linked to improve colloidal stability. Monoclonal antibodies against vascular endothelial growth factor (mAbVEGF) were covalently conjugated to BSA through a polyethyleneglycol linker. Here we demonstrate that 1) BSA coated nanoparticles are stable and non-toxic to different cells at concentration up to 2.5mg/mL; 2) conjugation of monoclonal antibodies to nanoparticles promotes their binding to VEGF-positive glioma С6 cells in vitro; 3) targeted nanoparticles are effective in MRI visualization of the intracranial glioma. Thus, mAbVEGF-targeted BSA-coated magnetic nanoparticles are promising MRI contrast agents for glioma visualization. FROM THE CLINICAL EDITOR This work focuses on synthesis and characterization of targeted magnetic nanoparticles as magnetic resonance imaging (МRI) agents for in vivo visualization of gliomas. The authors utilize the fact that high-grade gliomas have extensive areas of necrosis and hypoxia, which results in increased secretion of angiogenesis vascular endothelial growth factor (VEGF). Monoclonal antibodies against vascular endothelial growth factor (mAbVEGF) were covalently conjugated to crosslinked BSA coated ferric oxide (Fe3O4) nanoparticles. The results show that these targeted nanoparticles are effective in MRI visualization of the intracranial glioma and may provide a new and promising contrast agent.

Targeted delivery of liposomal nanocontainers to the peritumoral zone of glioma by means of monoclonal antibodies against GFAP and the extracellular loop of Cx43.

UNLABELLED The selectivity of PEGylated immunoliposomes based on monoclonal antibodies against GFAP and the E2 extracellular loop of connexin 43 (MAbE2Cx43) with respect to the focus of a glioma was estimated in experiments on animals with intracranial C6 glioma. Stealth immunoliposomes were labeled with 2 alternative labels, a fluorescent (Dil C18) and a paramagnetic (Gd-DTPA) one. Fluorescent-labeled liposomal nanocontainers were detected at the periphery of the glioma, where the target antigens were overexpressed, 48 hours after injection. Dynamic T1 MRI of rats injected with paramagnetic immunoliposomes carrying MAbE2Cx43 showed distinct accumulation of the paramagnetic contrast agent at the periphery of the glioma, which began 6 hours after administration. These data suggest that immunoliposomal nanocontainers based on antibodies against GFAP and the E2 extracellular fragment of connexin 43 are suitable for targeted delivery of diagnostic and therapeutic drugs to the peritumoral invasion zone of high-grade gliomas. FROM THE CLINICAL EDITOR PEGylated immunoliposomes based on monoclonal antibodies against GFAP and the E2 extracellular loop of connexin 43 were investigated in animals with intracranial C6 glioma. These immunoliposomal nanocontainers were found suitable for targeted delivery of diagnostic and therapeutic drugs to the peritumoral invasion zone of high-grade gliomas.

PEGylated immunoliposomes directed against brain astrocytes.

Polyethylene glycol (PEG)ylated (stealth) immunoliposomes directed against human gliofibrillary acidic protein (GFAP) were prepared by coupling the thiolated monoclonal anti-GFAP antibodies with a maleimide derivative of phosphatidyl ethanolamine of the liposomal membrane. Experiments with cell cultures demonstrated specific and competitive binding of these immunoliposomes to embryonic rat brain astrocytes. Administered intravenously into rats, the immunoliposomes displayed typical kinetics with elimination half-lives of 8–15 hr. Being incapable of penetrating the unimpaired blood-brain barrier (BBB), these immunoliposomes, nevertheless, may be useful in delivering drugs to glial brain tumors (which continue to express GFAP) or to other pathological loci in the brain with a partially disintegrated BBB.

Modeling and immunohistochemical analysis of C6 glioma in vivo.

A reproducible in vivo model of C6 glioma was developed in Wistar rats. Analysis of histological preparations showed similar morphology of rat C6 glioma and human glioblastoma. The formation of a glial border at the periphery of the glioma, consisting of GFAP-positive reactive astrocytes, was shown by the immunohistochemical method. The border appeared on day 8 after implantation, astrogliosis was observed until animal death (day 28). Reactive astrocytes with branched processes surrounded not only the primary glioma focus, but also all sites of tumor invasion in the nervous tissue. Expression of EBA (blood-brain barrier marker) was disturbed and synthesis of AMVB1 (endothelial antigen) increased in neoplastic endotheliocytes, which suggested pronounced functional restructuring of the blood-tumor barrier in comparison with the blood-brain barrier. The phenomenon of predominant expression of GFAP and AMVB1 in the tumor tissue can be used for the development of systems for targeted drug transport into the tumor by means of appropriate antibodies.