Focused ultrasound delivery of a selective TrkA agonist rescues cholinergic function in a mouse model of Alzheimer's disease.

The degeneration of cholinergic neurons is a prominent feature of Alzheimer's disease (AD). In animal models of injury and aging, nerve growth factor (NGF) enhances cholinergic cell survival and function, contributing to improved memory. In the presence of AD pathology, however, NGF-related therapeutics have yet to fulfill their regenerative potential. We propose that stimulating the TrkA receptor, without p75NTR activation, is key for therapeutic efficacy. Supporting this hypothesis, the selective TrkA agonist D3 rescued neurotrophin signaling in TgCRND8 mice, whereas NGF, interacting with both TrkA and p75NTR, did not. D3, delivered intravenously and noninvasively to the basal forebrain using MRI-guided focused ultrasound (MRIgFUS)-mediated blood-brain barrier (BBB) permeability activated TrkA-related signaling cascades and enhanced cholinergic neurotransmission. Recent clinical trials support the safety and feasibility of MRIgFUS BBB modulation in AD patients. Neuroprotective agents targeting TrkA, combined with MRIgFUS BBB modulation, represent a promising strategy to counter neurodegeneration in AD.

Reduced in vivo lung metastasis of a breast cancer cell line after treatment with Herceptin mAb conjugated to chemotherapeutic drugs.

Anthracyclines and taxanes have remarkable anticancer efficacy, but have poor selectivity and high toxicity. Targeted delivery of chemotherapeutics has emerged as a strategy to achieve higher drug levels at the tumor site, to spare noncancerous tissue and potentially to use lower systemic drug doses, thus preventing side effects. In this study, we targeted the HER2 receptor using the monoclonal antibody (mAb) Herceptin (Trastuzumab) chemically conjugated to Doxorubicin or Taxol. In vitro, drug-Herceptin conjugates exhibited cytotoxicity comparable to equimolar concentrations of free drugs, with the benefit that the cytotoxicity of the conjugates was selective for cells expressing the HER2 target. In vivo, treatment of tumor-bearing mice with Taxol-Herceptin conjugates had a reduction of primary tumors comparable to equivalent doses of free drugs. However, Taxol-Herceptin conjugates significantly reduced metastasis compared with equivalent doses of free drugs. Thus, the data support the concept that conjugates might target metastasis better than primary tumors. This would offer a potential therapeutic approach for management of metastatic breast cancer.

VRQ397 (CRAVKY): a novel noncompetitive V2 receptor antagonist.

Vasopressin type 2 receptor (V2R) exhibits mostly important properties for hydroosmotic equilibrium and, to a lesser extent, on vasomotricity. Drugs currently acting on this receptor are analogs of the natural neuropeptide, arginine vasopressin (AVP), and hence are competitive ligands. Peptides that reproduce specific sequences of a given receptor have lately been reported to interfere with its action, and if such molecules arise from regions remote from the binding site they would be anticipated to exhibit noncompetitive antagonism, but this has yet to be shown for V2R. Six peptides reproducing juxtamembranous regions of V2R were designed and screened; the most effective peptide, cravky (labeled VRQ397), was characterized. VRQ397 was potent (IC(50) = 0.69 +/- 0.25 nM) and fully effective in inhibiting V2R-dependent physiological function, specifically desmopressin-L-desamino-8-arginine-vasopressin (DDAVP)-induced cremasteric vasorelaxation; this physiological functional assay was utilized to avoid overlooking interference of specific signaling events. A dose-response profile revealed a noncompetitive property of VRQ397; correspondingly, VRQ397 bound specifically to V2R-expressing cells could not displace its natural ligand, AVP, but modulated AVP binding kinetics (dissociation rate). Specificity of VRQ397 was further confirmed by its inability to bind to homologous V1 and oxytocin receptors and its inefficacy to alter responses to stimulation of these receptors. VRQ397 exhibited pharmacological permissiveness on V2R-induced signals, as it inhibited DDAVP-induced PGI(2) generation but not that of cAMP or recruitment of beta-arrestin2. Consistent with in vitro and ex vivo effects as a V2R antagonist, VRQ397 displayed anticipated in vivo aquaretic efficacy. We hereby describe the discovery of a first potent noncompetitive antagonist of V2R, which exhibits functional selectivity, in line with properties of a negative allosteric modulator.

Reactions of CH(3)CHO(.+) and of CH(3)COH(.+) with water upon Fourier transform ion cyclotron resonance conditions.

The reactions of CH(3)CHO(+) and of CH(3)COH(+) with water yield the same products, at almost the same rate. It is shown, by using a characteristic reaction of the carbene structure, that a molecule of water converts CH(3)COH(+) into its more stable isomer CH(3)CHO(+), which is a new example of catalyzed 1,2-H transfer. The dominant product is the proton-bound dimer of water which, in fact, comes from the [H(2)OH(+)...CH(3)(.)] and [H(2)OH(+)...CO] primary products whose observed abundances are poor. In a related system, ionized formamide/water, a water molecule catalyzes the 1,3-transfer leading from the solvated carbene to the [H(2)O...H(+)...H(2)N-C=O)] stable intermediate, which eliminates CO without back energy. In contrast, such a process does not take place in the studied system since the cleavage of the so formed [H(2)OH(+)...CH(3)CO] transient intermediate involves a high back energy; this is explained by the charge repartition within this intermediate. In fact, a different pathway takes place. The solvated acetaldehyde ion isomerizes into a terbody intermediate in which protonated water is bonded to a CO molecule on the one hand and to a methyl radical on the other hand. Simple cleavages of this complex yield the observed products.

The

The bimolecular reaction of the CH2CHOH+ enol ion (m/z 44) with acetaldehyde gives a strongly dominant product, m/z 45, formed mainly by proton transfer from the ion to the molecule. The abundance of the product coming from a H* abstraction reaction from the neutral, albeit more exothermic, is negligible. In order to explain this result, the long lived [CH2CHOH*+, CH3CHO] solvated ion was generated by reaction of the CH2CHOH*+ enol ion with (CH3CHO)n in the cell of a Fourier transform ion cyclotron resonance mass spectrometer. The structure of this solvated ion was clearly established. Labeling indicates that [CH2CHOH+, CH3CHO], upon low energy collisions, reacts by H* abstraction more rapidly than by H+ transfer to the neutral moiety. This shows that the entropic factors are determinant when the enol ion reacts directly with acetaldehyde.

Opiate-like peptides. Part XI. 2-[2-phenyl-1,3-indandionyl]--amides of enkephalin analogs. Synthesis and analgesic activity.

Four enkephalin analogs containing C-terminal 2-amino-2-phenyl-1,3-indandione residue were prepared: [Met-NHPID5]--enkephalin (E3), [D-Ala2,Met-NHPID5]--enkephalin (E4), [Leu-NHPID5]--enkephalin (E5), [D-Ala2,Leu-NHPID5]--enkephalin (E6). Their analgesic activities were assayed by three in vivo tests: the "hot plate" method, the reaction to electric painful stimulus and the writhing syndrome test. Neurotoxicity effects were determined by the rota-rod test. The most pronounced analgesic effect was induced by compounds with D-Ala in position 2, particularly in the "hot plate" test.