Phase I study of sorafenib and tipifarnib for recurrent glioblastoma: NABTC 05-02.

Recurrent glioblastoma (GBM) has a very low 6-month progression free survival (PFS) with currently available treatments. Combination chemotherapy to target multiple cell signaling pathways is currently being investigated in order to improve prognosis for recurrent disease. The purpose of this phase I study was to determine the maximum tolerated dose (MTD) for the combination of tipifarnib and sorafenib for the treatment of recurrent GBM. Patients with pathologically proven WHO grade IV GBM and radiographically proven tumor recurrence were eligible for this study. Treatments included sorafenib at twice daily and escalating dosages of tipifarnib. Dose-limiting toxicity (DLT) was determined over the first 28-days of treatments, and the MTD was determined in a 3 + 3 study design. We enrolled 24 patients, and 21 patients completed the MTD period. The study was stopped early with no MTD determination for excessive toxicities. The last dose level reached was sorafenib at 200 mg twice a day and tipifarnib 100 mg twice a day on an alternating week schedule. The DLTs included diarrhea, lipase elevation, hypophosphatemia, and arthralgia. The combination of sorafenib and tipifarnib has excessive toxicities and full single agent dosages could not be achieved in combination.

A randomized phase II of gemcitabine and sorafenib versus sorafenib alone in patients with metastatic pancreatic cancer.

PURPOSE: Patients with metastatic pancreatic cancer have limited therapeutic options. The role of the Ras-Raf-MAPK pathway and of vascular endothelial growth factor in pancreatic carcinogenesis provided the rational to evaluate the efficacy of sorafenib with or without gemcitabine in a randomized phase II study. METHODS: Patients with metastatic pancreatic cancer were randomized to sorafenib alone (arm A) or sorafenib with gemcitabine (arm B). RESULTS: Arm A was closed to accrual at interim analysis due to the lack of objective response. Median PFS and OS were 2.3 and 4.3 months respectively. There was one partial response among the 37 patients in arm B. Median PFS and OS were 2.9 and 6.5 months respectively. There were more grade 3 and 4 toxicities in arm B with the most common being neutropenia (17%), thrombocytopenia (8%), alkaline phosphatase elevation (14%), venous thromboembolism (8%), diarrhea, hypokalemia and ALT elevation (5%) each. Several associations were noted between single nucleotide polymorphisms in ribonucleotide reductase, Cox-2, vascular endothelial growth factor and survival in patients treated with gemcitabine and sorafenib. CONCLUSIONS: Neither sorafenib alone or sorafenib in combination with gemcitabine manifested promising activity in metastatic pancreatic cancer.

A pharmacodynamic study of sorafenib in patients with relapsed and refractory acute leukemias.

We report the results of a phase I dose escalation trial of the multikinase inhibitor sorafenib in relapsed and refractory acute leukemia patients using an intermittent dosing regimen. Fifteen patients with advanced leukemia (12 with acute myeloid leukemia, 2 with acute lymphoblastic leukemia, 1 with biphenotypic) and a median age of 63 (range 37-85) years were enrolled and treated on a dose escalation trial. Toxicities >or=grade 3 were present in 55% of cycles and the maximum tolerated dose (MTD) was determined to be 400 mg b.i.d. x 21 days in a 28-day cycle. Plasma inhibitory assays of kinase targets extracellular signal-regulated kinase (ERK) and FLT3-internal tandem duplication (ITD) showed excellent target inhibition, with FLT3-ITD silencing occurring below the MTD. The N-oxide metabolite of sorafenib seemed to be a more potent inhibitor of FLT3-ITD than the parent compound. Despite marked ex vivo FLT-3 ITD inhibition, no patients met the criteria for complete or partial response in this monotherapy study. Out of 15 patients, 11 experienced stable disease as best response. Although sorafenib showed only modest clinical activity as a single agent in this heavily treated population, robust inhibition of FLT3 and ERK suggests that there may be a potential important role in combination therapies.

Measurement of phase gradients in the EEG.

Previous research has shown that spatio-temporal waves in the EEG are generally of long spatial wavelength and form smooth patterns of phase gradients at particular time-samples. This paper describes a method to measure smooth phase gradients of long spatial wavelength in the EEG. The method depends on the global pattern of phase at a given frequency and time and is therefore robust to variations, over time, in phase-lag between particular sites. Phases were estimated in the EEG signal using wavelet or short time-series Fourier methods. During an auditory oddball task, phases across the scalp tend to fall within a limited circular range, a range that is not indicative of phase-synchrony nor waves with multiple periods. At times the phases tended to maintain a spatially and temporally ordered relationship. The relative phases were analysed using three phase gradient basis functions, providing a measure of the amount of variance explained, across the electrodes, by smooth changes in relative phase from a single minimum or single maximum. The data from 586 adult subjects were analysed and it was found that the probability of phase gradient events varies with time and frequency in the stimulus-locked average, and with task demands. The temporal extent of spatio-temporal waves was measured by detecting smoothly changing patterns of phase latencies across the scalp. The specific spatial pattern and timing of phase gradients correspond closely to the latency distributions of certain ERPs.

Neurophysical modeling of brain dynamics.

A recent neurophysical model of brain electrical activity is outlined and applied to EEG phenomena. It incorporates single-neuron physiology and the large-scale anatomy of corticocortical and corticothalamic pathways, including synaptic strengths, dendritic propagation, nonlinear firing responses, and axonal conduction. Small perturbations from steady states account for observed EEGs as functions of arousal. Evoked response potentials (ERPs), correlation, and coherence functions are also reproduced. Feedback via thalamic nuclei is critical in determining the forms of these quantities, the transition between sleep and waking, and stability against seizures. Many disorders correspond to significant changes in EEGs, which can potentially be quantified in terms of the underlying physiology using this theory. In the nonlinear regime, limit cycles are often seen, including a regime in which they have the characteristic petit mal 3 Hz spike-and-wave form.

Unified neurophysical model of EEG spectra and evoked potentials.

Evoked potentials -- the brain's transient electrical responses to discrete stimuli -- are modeled as impulse responses using a continuum model of brain electrical activity. Previous models of ongoing brain activity are refined by adding an improved model of thalamic connectivity and modulation, and by allowing for two populations of excitatory cortical neurons distinguished by their axonal ranges. Evoked potentials are shown to be modelable as an impulse response that is a sum of component responses. The component occurring about 100 ms poststimulus is attributed to sensory activation, and this, together with positive and negative feedback pathways between the cortex and thalamus, results in subsequent peaks and troughs that semiquantitatively reproduce those of observed evoked potentials. Modulation of the strengths of positive and negative feedback, in ways consistent with psychological theories of attentional focus, results in distinct responses resembling those seen in experiments involving attentional changes. The modeled impulse responses reproduce key features of typical experimental evoked response potentials: timing, relative amplitude, and number of peaks. The same model, with further modulation of feedback, also reproduces experimental spectra. Together, these results mean that a broad range of ongoing and transient electrocortical activity can be understood within a common framework, which is parameterized by values that are directly related to physiological and anatomical quantities.

Novel mimics of sialyl Lewis X: design, synthesis and biological activity of a series of 2- and 3-malonate substituted galactoconjugates.

A series of potent inhibitors of P-selectin as potential anti-inflammatory agents is reported. These compounds are derivatives of galactocerebrosides bearing a malonate side chain in positions 2 and 3 of the galactose moiety. Based on the binding mode of sialyl Lewis X, the two acidic groups of the malonate are designed to form ionic interactions with two important lysines in the active site of P-selectin, Lys113 and Lys111. On the other hand, the 4- and 6-hydroxy groups on the galactose ring are arranged to chelate the calcium ion in the P-selectin active site. The synthesis and the biological activity of this series of compounds are described. Lead compounds having a greater potency than sialyl Lewis X are identified.

Prediction of electroencephalographic spectra from neurophysiology.

A recent neurophysical model of propagation of electrical waves in the cortex is extended to include a physiologically motivated subcortical feedback loop via the thalamus. The electroencephalographic spectrum when the system is driven by white noise is then calculated analytically in terms of physiological parameters, including the effects of filtering of signals by the cerebrospinal fluid, skull, and scalp. The spectral power at low frequencies is found to vary as f(-1) when awake and f(-3) when asleep, with a breakpoint to a steeper power-law tail at frequencies above about 20 Hz in both cases; the f(-1) range concurs with recent magnetoencephalographic observations of such a regime. Parameter sensitivities are explored, enabling a model with fewer free parameters to be proposed, and showing that spectra predicted for physiologically reasonable parameter values strongly resemble those observed in the laboratory. Alpha and beta peaks seen near 10 Hz and twice that frequency, respectively, in the relaxed wakeful state are generated via subcortical feedback in this model, thereby leading to predictions of their frequencies in terms of physiological parameters, and of correlations in their occurrence. Subcortical feedback is also predicted to be responsible for production of anticorrelated peaks in deep sleep states that correspond to the occurrence of theta rhythm at around half the alpha frequency and sleep spindles at 3/2 times the alpha frequency. An additional positively correlated waking peak near three times the alpha frequency is also predicted and tentatively observed, as are two new types of sleep spindle near 5/2 and 7/2 times the alpha frequency, and anticorrelated with alpha. These results provide a theoretical basis for the conventional division of EEG spectra into frequency bands, but imply that the exact bounds of these bands depend on the individual. Three types of potential instability are found: one at zero frequency, another in the theta band at around half the alpha frequency, and a third at the alpha frequency itself.

Molecular heterogeneity and function of EWS-WT1 fusion transcripts in desmoplastic small round cell tumors.

Desmoplastic small round cell tumor (DSRCT) is a primitive sarcoma with a consistent cytogenetic abnormality, t(11;22)(p13;q12). This chromosomal translocation generates a chimeric transcript that is formed by fusion of the 5' region of the Ewing's sarcoma gene, EWS, with the 3' DNA-binding segment of WT1, the Wilms' tumor suppressor gene. We collected 14 DSRCT tumor samples and examined the hybrid transcripts. We identified: (a) combinatorial heterogeneity of EWS exons fused to WT1 including use of EWS exons 7, 8, and 9; (b) subpopulations of variant transcripts in 6 of 14 tumors characterized by aberrant splicing resulting in loss of EWS exon 6 or WT1 exon 9; (c) multiple cDNA products with large internal deletions; and (d) insertion of small stretches of heterologous DNA at the fusion site or exon splice region in transcripts from two tumors. Most of the splice variants were in-frame, and in vitro translated fusion proteins with intact DNA-binding motifs formed complexes with a WT1 response element in gel mobility assays. Each of the chimeric proteins retains the ability to bind to the GC and TC elements of the early transcription factor EGR-1 as well as WT1 consensus sequences. We present evidence that various EWS-WT1 proteins up-regulated EGR-1 promoter activity and that this up-regulation is specifically dependent upon the absence of the exon 9 KTS domain of WT1. The molecular diversity and functionality exhibited by these fusion transcripts may have significant biological implications for their transactivating and tumorigenic potential.

Synchronous cortical gamma-band activity in task-relevant cognition.

Widespread synchronous oscillatory activity, particularly in the gamma ('40 Hz') band, has been postulated to exist in the brain as a mechanism underlying binding. A new method of examining phase synchronicity across multiple electrode sites in specific EEG frequency bands as a function of time was employed, in a conventional cognitive ERP paradigm in 40 normal subjects. A significant late post-stimulus gamma synchronicity response occurred for task-relevant stimuli, whereas for task-irrelevant stimuli no such response was evident. However, an early response was seen for both task-relevant and irrelevant stimuli. This is the first empirical demonstration that widespread synchronous high frequency oscillations occur in humans in relation to cognition.

Response of hypercholesterolemic subjects to administration of tocotrienols.

The cholesterol-suppressive actions of Palmvitee and gamma-tocotrienol were assessed in hypercholesterolemic subjects after acclimation to the American Heart Association Step 1 dietary regimen for four and eight weeks, respectively. The four-week dietary regimen alone elicited a 5% decrease (P < 0.05) in the cholesterol level of the 36 subjects. Subjects continuing on the dietary regimen for a second four-week period experienced an additional 2% decrease in their cholesterol levels. Dietary assessments based on unanticipated recalls of 24-h food intake records suggest that significant reductions in energy and fat, predominantly in saturated fat, intakes are responsible. The subjects experienced significant Palmvitee- and gamma-tocotrienol-mediated decreases in cholesterol. The group of subjects acclimated to the dietary regimen for four weeks responded to Palmvitee (a blend of tocols providing 40 mg alpha-tocopherol, 48 mg alpha-tocotrienol, 112 mg gamma-tocotrienol, and 60 mg delta-to-cotrienol/day for four weeks) with a 10% decrease in cholesterol (P < 0.05). Dietary assessments showed no further change in energy and fat intakes. alpha-Tocopherol attenuated the cholesterol-suppressive action of the tocotrienols. The second group of subjects, acclimated to the dietary regimen for eight weeks, received 200 mg gamma-tocotrienol/d for four weeks. The cholesterol-suppressive potency of this alpha-tocopherol-free preparation was calculated to be equivalent to that of the mixture of tocotrienols (220 mg) used in the prior study. Cholesterol levels of the 16 subjects in the second group decreased 13% (P < 0.05) during the four-week trial. Plasma apolipoprotein B and ex vivo generation of thromboxane B2 were similarly responsive to the tocotrienol preparations, whereas neither preparation had an impact on high density lipoprotein cholesterol and apolipoprotein A-1 levels.

Inverse filter computation of the neural impulse giving rise to the auditory evoked potential.

An impulse response hypothesis for evoked potentials is tested. The auditory evoked potential (AEP) is shown to be the consequence of an impulse (the arrival of sensory signals in cortex) giving rise to an impulse response (the resonation of electrocortical activity in the form of group linear waves). To demonstrate this, pre- and post-stimulus EEG activity was recorded from subjects engaged in performance of an auditory odd-ball experiment. For each stimulus, the impulse required to account for the single auditory evoked potential (AEP) as a linear impulse response, was computed by use of the inverse of a filter obtained by autoregression analysis of the pre-stimulus EEG epoch. Single estimations of the impulse were then averaged. The average impulse exhibits a time course and topology consistent with the arrival of neural volleys in the cortex. The physical validity of the hypothesis is supported by a high lag correlation of the following values of the AEP to the average impulse. A further test calculation supports the linear additivity assumptions of the hypothesis.

Gross electrocortical activity as a linear wave phenomenon with variable temporal damping, regulated by ascending catecholamine neurones.

We report critical tests for a theory of electrocortical wave processes, in which telencephalic dendritic potentials reflect the mass action of coupled oscillatory circuits exhibiting complicated and unspecified non-linearities, the whole system being driven by active cell firing. Specific assumptions were: stochastic independence for instantaneous coupling parameters in the system, an individual central tendency to the cycle time for each circuit, and the maintenance of steady state conditions. Application of the central limit theorem to the state transition matrix shows that the gross electrocortical waves should be linear waves, exhibiting a multitude of invariant resonant modes, with the natural frequencies of all the modes being clustered about a smaller number of center values. Ascending brain-stem neurones of at least the dopaminergic and noradrenergic classes should regulate both the power of noise-like signals driving the telencephalic resonant patterns, and the temporal damping of each resonance. We devised tests which involved between hemisphere comparisons of electrocortical spectra, before and after unilateral lesion of transhypothalamic ascending fibres, thus obtaining ratio power changes attributable to post lesion asymmetry of damping and driving, in modes of equivalent left-right center-frequencies. These ratio spectra were curve-fitted to an approximate theoretical expression, and the parameters obtained enabled tests of several specific predictions. Estimates of the center values for resonant mode frequencies, comparison of the relative changes in left/right phase with that expected from the ratio changes in power, and estimates of the surface-to-signal transformation of left and right signals made by a back-calculation, all conform to expectation from the theory, and are consistent across lesion of different types of ascending neurone.

State-changes in the brain viewed as linear steady-states and non-linear transitions between steady-states.

Preceding papers concern a linear theory of electrocortical waves and their regulation by brain-stem neurones, in conditions of steady-state. The present paper reconsiders the theory, and generalizes beyond the effects of the fibre systems so far studied. Relaxation and unification of the assumptions upon which the initial model was based is undertaken. It is shown that the generalised model may render state changes within the brain accessible to systematic description, using the EEG as dependent variable. It is proposed that a multitude of stable states are possible within the brain, each characterised by a set of damping parameters for separate linear resonant modes. Within each stable state, the set of sums of resonant modes characterises a sub-space of the total state-space. Transition between stable regions can occur with either perturbation by external signals, or by internal controls. Tentative consideration is given to the role of plastic changes leading to adaptive learning as an attribute of a system of this type.

Amplitude and phase relations of electrocortical waves regulated by transhypothalamic dopaminergic neurones: a test for a linear theory.

We have previously proposed that electrocortical activity (EEG) arises as a manifestation of linear waves generated by resonance among telencephalic neurones, and that this activity is controlled in part by ascending neurones from the brain-stem, which regulate the damping of each resonance. The present experiments focus on a specific class of ascending neurones, the mesotelencephalic dopaminergic cells, because these cells are thought to mediate important psychological effects, and are conveniently subject to selective lesion. A critical test of the theory is undertaken, by performing selective unilateral lesion, assessing the changes in the power spectrum of the EEG attributable to lesion, and determining whether the changes in phase of the EEG correspond to that predicted from the changes in power. Results support the theory, although the model order applicable in these experiments in inadequate. The consequences of these findings for automata theory, linear network theory and their application to mammalian brains are briefly discussed.

A preliminary mathematical model for lateral hypothalamic regulation of electrocortical activity.

The assumption that transhypothalamic neurones exert damping effects on electrocortical oscillatory activity enables a preliminary model to be advanced, in which the action of the lateral hypothalamus upon the EEG is that of a linear filter. Predictions of the model were tested on data obtained from lateral hypothalamic lesion and stimulation studies.

Lateral hypothalamic regulation of electrocortical activity.

The EEG power spectrum of normal and split-brain cats was analysed before and after unilateral hypothalamic lesion at sites which prior to lesion had ICSS or aversive properties on stimulation, and which gave rise to sensorimotor neglect after lesion. A filter-gain function was calculated to describe the lesions' effect upon the ipsilateral EEG spectrum in stable conditions, and was found to be approximated by a specific gamma function. This partially defines the action that the lateral hypothalamus may exert on electrocortical activity.