Therapeutic melanoma inhibition by local micelle-mediated cyclic nucleotide repression

The acidic tumor microenvironment in melanoma drives immune evasion by up-regulating cyclic adenosine monophosphate (cAMP) in tumor-infiltrating monocytes. Here we show that the release of non-toxic concentrations of an adenylate cyclase (AC) inhibitor from poly(sarcosine)-block-poly(L-glutamic acid γ-benzyl ester) (polypept(o)id) copolymer micelles restores antitumor immunity. In combination with selective, non-therapeutic regulatory T cell depletion, AC inhibitor micelles achieve a complete remission of established B16-F10-OVA tumors. Single-cell sequencing of melanoma-infiltrating immune cells shows that AC inhibitor micelles reduce the number of anti-inflammatory myeloid cells and checkpoint receptor expression on T cells. AC inhibitor micelles thus represent an immunotherapeutic measure to counteract melanoma immune escape.

Efficacy of florpyrauxifen-benzyl for eurasian watermilfoil control and nontarget Illinois pondweed, elodea, and coontail response

This research evaluated low concentrations and short exposure times of the recently registered aquatic herbicide florpyrauxifen-benzyl (4-amino-3-chloro-6-(4-chloro-2-fluoro-3-methoxyphenyl)-5-fluoro-pyridine-2-benzyl ester) on the target plant Eurasian watermilfoil (Myriophyllum spicatum L., hereafter referred to as EWM) as well as selectivity towards the nontarget submersed species Illinois pondweed (Potamogeton illinoensis Morong), elodea (Elodea canadensis Michx.), and coontail (Ceratophyllum demersum L.)

Synergy of Hydeal-D® and Hyaluronic Acid for Protecting and Restoring Urothelium: In Vitro Characterization

Interstitial cystitis (IC) or painful bladder syndrome is a chronic dysfunction due to an inflammatory condition, characterized by bladder pain and urinary frequency. Currently, no gold standard therapy is available since IC does not respond to conventional ones. Given these premises, the aim of this work was the in vitro characterization of biological properties (mucoadhesion and anti-inflammatory activity) of a commercial product (HydealCyst–HydC) based on hyaluronic acid (HA) and the benzyl ester of HA (Hydeal-D®) intended for bladder instillation to restore and/or protect the urothelial layer of glycosamino glycans (GAGs). The in vitro characterization demonstrated that an interaction product is formed between HA and Hydeal-D® that has a role in the rheological behavior and mucoadhesive properties. HA was identified as a key component to form the mucoadhesive joint, while the interaction of HA with Hydeal-D® improved polysaccharide stability and prolonged the activity ex vivo. Moreover, HydC is cytocompatible with urothelial cells (HTB-4) and possesses an anti-inflammatory effect towards these cells by decreasing the secretion of IL-6 and IL-8, which were both increased in patients with IC, and by increasing the secretion of sulfated GAGs. These two findings, along with the resilience properties of the formulation due to mucoadhesion, suggest the active role of HydC in protecting and restoring urothelium homeostasis.

Aminated Graphene-Graft-Oligo(Glutamic Acid) /Poly(ε-Caprolactone) Composites: Preparation, Characterization and Biological Evaluation

Biodegradable and biocompatible composites are of great interest as biomedical materials for various regeneration processes such as the regeneration of bones, cartilage and soft tissues. Modification of the filler surface can improve its compatibility with the polymer matrix, and, as a result, the characteristics and properties of composite materials. This work is devoted to the synthesis and modification of aminated graphene with oligomers of glutamic acid and their use for the preparation of composite materials based on poly(ε-caprolactone). Ring-opening polymerization of N-carboxyanhydride of glutamic acid γ-benzyl ester was used to graft oligomers of glutamic acid from the surface of aminated graphene. The success of the modification was confirmed by Fourier-transform infrared and X-ray photoelectron spectroscopy as well as thermogravimetric analysis. In addition, the dispersions of neat and modified aminated graphene were analyzed by dynamic and electrophoretic light scattering to monitor changes in the characteristics due to modification. The poly(ε-caprolactone) films filled with neat and modified aminated graphene were manufactured and carefully characterized for their mechanical and biological properties. Grafting of glutamic acid oligomers from the surface of aminated graphene improved the distribution of the filler in the polymer matrix that, in turn, positively affected the mechanical properties of composite materials in comparison to ones containing the unmodified filler. Moreover, the modification improved the biocompatibility of the filler with human MG-63 osteoblast-like cells.

Conjugation of Diclofenac with Novel Oleanolic Acid Derivatives Modulate Nrf2 and NF-κB Activity in Hepatic Cancer Cells and Normal Hepatocytes Leading to Enhancement of Its Therapeutic and Chemopreventive Potential

Combining NSAIDs with conventional therapeutics was recently explored as a new strategy in cancer therapy. Our earlier studies showed that novel oleanolic acid oximes (OAO) conjugated with aspirin or indomethacin may enhance their anti-cancer potential through modulation of the Nrf2 and NF-κB signaling pathways. This study focused on the synthesis and biological evaluation of four diclofenac (DCL)–OAO derivative conjugates in the context of these pathways’ modification and hepatic cells survival. Treatment with the conjugates 4d, 3-diclofenacoxyiminoolean-12-en-28-oic acid morpholide, and 4c, 3-diclofenacoxyiminoolean-12-en-28-oic acid benzyl ester significantly reduced cell viability in comparison to the DCL alone. In THLE-2, immortalized normal hepatocytes treated with these conjugates resulted in the activation of Nrf2 and increased expression in SOD-1 and NQO1, while the opposite effect was observed in the HepG2 hepatoma cells. In both cell lines, reduced activation of the NF-κB and COX-2 expression was observed. In HepG2 cells, conjugates increased ROS production resulting from a reduced antioxidant defense, induced apoptosis, and inhibited cell proliferation. In addition, the OAO morpholide derivative and its DCL hybrid reduced the tumor volume in mice bearing xenografts. In conclusion, our study demonstrated that conjugating diclofenac with the OAO morpholide and a benzyl ester might enhance its anti-cancer activity in HCC.

Activity of Compounds from Temperate Propolis against Trypanosoma brucei and Leishmania mexicana

Ethanolic extracts of samples of temperate zone propolis, four from the UK and one from Poland, were tested against three Trypanosoma brucei strains and displayed EC50 values < 20 µg/mL. The extracts were fractionated, from which 12 compounds and one two-component mixture were isolated, and characterized by NMR and high-resolution mass spectrometry, as 3-acetoxypinobanksin, tectochrysin, kaempferol, pinocembrin, 4′-methoxykaempferol, galangin, chrysin, apigenin, pinostrobin, cinnamic acid, coumaric acid, cinnamyl ester/coumaric acid benzyl ester (mixture), 4′,7-dimethoxykaempferol, and naringenin 4′,7-dimethyl ether. The isolated compounds were tested against drug-sensitive and drug-resistant strains of T. brucei and Leishmania mexicana, with the highest activities ≤ 15 µM. The most active compounds against T. brucei were naringenin 4′,7 dimethyl ether and 4′methoxy kaempferol with activity of 15–20 µM against the three T. brucei strains. The most active compounds against L. mexicana were 4′,7-dimethoxykaempferol and the coumaric acid ester mixture, with EC50 values of 12.9 ± 3.7 µM and 13.1 ± 1.0 µM. No loss of activity was found with the diamidine- and arsenical-resistant or phenanthridine-resistant T. brucei strains, or the miltefosine-resistant L. mexicana strain; no clear structure activity relationship was observed for the isolated compounds. Temperate propolis yields multiple compounds with anti-kinetoplastid activity.

Creatine benzyl ester is effective for prevention and treatment of neurological and cognitive disorders caused by focal cerebral ischemia in rats

Введение. Ишемический инсульт головного мозга является одной из главных причин преждевременных смертей во всем мире, вызванные им когнитивные и функциональные нарушения ведут к инвалидизации. Для клинического использования в настоящее время одобрено лишь одно средство - рекомбинантный активатор плазминогена, который обеспечивает восстановление (реканализацию) мозгового кровотока. Средства эффективной нейропротекции, обеспечивающие непосредственную защиту нейронов от ишемии (предотвращение апоптоза, сохранение функциональной активности), несмотря на огромное число исследований, по-прежнему не найдены. Поиск новых подходов к профилактике и лечению острых нарушений мозгового кровообращения по-прежнему остается одной из наиболее актуальных проблем современной медицины. Цель исследования - изучение влияния бензилового эфира креатина, нового синтетического соединения, обладающего способностью проникать через гематоэнцефалический барьер и осуществлять нейропротекторное действие на неврологические и когнитивные нарушения, вызванные фокальной ишемией головного мозга у крыс линии Спрэг-Доули. Методика. Фокальную ишемию головного мозга (ФИМ) индуцировали путем хронической окклюзии левой средней мозговой артерии (СМА) в соответствии с модификацией методики A. Tamura и соавт., 1981. Использовали комбинированный наркоз: Золетил/Домитор (внутрибрюшинно (в/б) по 50 мг/кг и 0.2 мг/кг, соответственно), температуру тела поддерживали постоянной на уровне 38±0.5 oC. Окклюзию СМА производили путем биполярной высокочастотной электрокоагуляции (аппарат ЭХВЧ-25-11-С, «Медия», Россия) наиболее проксимального участка артерии, начиная с 0.5 мм от места ее отхождения от Виллизиева круга и на протяжении 2-3 мм. Процедура имитации ишемии (ложная операция) была аналогична вышеизложенной, но без электрокоагуляции СМА. Оценку степени неврологических нарушений проводили с использованием неврологической шкалы Гарсии и модифицированной шкалы тяжести неврологических нарушений (ОТНН) последовательно за 1 сут до и через 1, 3 и 7 сут после моделирования ФИМ. Для выявления когнитивных нарушений использовали водный лабиринт Морриса. Стандартный тест исследования процессов обучения и памяти применяли в версии для оценки пространственной референтной памяти, заключающейся в поиске животным скрытой под водой платформы при ориентации только по приметам окружающей обстановки. Бензиловый эфир креатина вводили внутрибрюшинно трижды - за 3, 2 и 1 ч до фокальной ишемии при профилактическом применении, либо через 1, 2 и 3 ч после моделирования ишемии при лечебном применении. Результаты. Получены приоритетные данные о том, что бензиловый эфир креатина при профилактическом введении эффективно предупреждает развитие неврологических нарушений и ослабляет когнитивные нарушения после фокальной ишемии мозга; при лечебном применении препарат ослабляет тяжесть неврологических нарушений и снижает выраженность когнитивных расстройств. Заключение. Результаты исследования позволяют рекомендовать данное производное креатина (после оценки возможной токсичности) для проведения первой фазы клинических испытаний в качестве препарата противоишемического действия. Background. Ischemic stroke is one of the main causes for premature death worldwide. Cognitive and functional disorders induced by stroke result in disability. Currently, only one agent has been approved for the clinical use, recombinant plasminogen activator, which provides recovery (recanalization) of cerebral blood flow. Despite numerous studies, an effective treatment to protect directly neurons from ischemia (preventing apoptosis, maintaining functional activity) has not been found so far. Thus, search for new approaches to prevention and treatment of acute cerebrovascular accidents remains one of the most pressing challenges of modern medicine. The aim of this work was to study the effect of creatine benzyl ester, a new synthetic compound capable for penetrating the blood-brain barrier to provide neuroprotection, on neurological and cognitive disorders induced by focal cerebral ischemia in Sprague-Dawley rats. Methods. Focal cerebral ischemia was induced by chronic occlusion of the left middle cerebral artery (MCA) by a A. Tamura et al. (1981) modified method. Combined intraperitoneal (i.p.) anesthesia with Zoletil/Domitor (50 mg/kg and 0.2 mg/kg, respectively) was used. Body temperature was maintained at 38±0.5 oC. MCA occlusion was performed by bipolar high-frequency electrocoagulation (EKVCh-25-11-C, Media, Russia) of the most proximal part of the artery, from 0.5 mm from the place where it originates from the circle of Willis and over 2-3 mm. The sham ischemia procedure was similar to that described above but without MCA electrocoagulation. Severity of neurological disorders was assessed with the Garcia neurological scale and a modified scale for severity of neurological disorders (OTTN) one day before and at 1, 3, and 7 days following ischemia. Cognitive impairment was assessed with the Morris water maze, a standard test for learning and memory. This test was used in a version for studying spatial reference memory, which consists of finding a platform hidden under the water by the animal guided only by surrounding marks. Creatine benzyl ester was injected i.p. three times, 3, 2 and 1 h before focal ischemia for prevention or 1, 2 and 3 h after ischemia for treatment. Results. The study produced priority data showing that preventive administration of creatine benzyl ester effectively abolished the development of neurological disorders and alleviated the cognitive impairment after focal cerebral ischemia whereas the therapeutic treatment restricted the severity of neurological and cognitive disorders. Conclusion. The results of the study supported recommendation of this creatine derivative (after assessing potential toxicity) for the first phase of clinical trials as an anti-ischemic drug.

Phenolic Compounds of Propolis Alleviate Lipid Metabolism Disorder

Lipid metabolism disorder is one of the significant risk factors for a multitude of human diseases and has become a serious threat to human health. The present study aimed to evaluate the effects of phenolics from poplar-type propolis on regulating lipid metabolism by using cell models of steatosis induced by palmitic acid (PA). Our study shows that phenolic esters have higher lipid-lowering activities than phenolic acids, especially for three caffeic acid esters, including caffeic acid phenethyl ester (CAPE), caffeic acid cinnamyl ester (CACE), and caffeic acid benzyl ester (CABE). Most notably, CACE presents prominent properties to prevent intracellular lipid accumulation and to amend extracellular adipokine secretion abnormalities. In addition, our results firstly reveal that CACE can alleviate lipid metabolism disorder through mediating protein kinase RNA-like endoplasmic reticulum kinase (PERK), activating transcription factor 6 (ATF6) signaling pathway-associated protein expression, suppressing endoplasmic reticulum (ER) stress, and activating peroxisome proliferator-activated receptors (PPARs) by distinct upregulation of PPARα and downregulation of PPARγ.