Synthesis and Characterization of Glycidyl Esters of Acrylic and Methacrylic Acids for NIPU Coatings

Glycidyl acrylate (GA) and Glycidylmethacrylate (GMA) are the reaction products of the epichlorohydrin (ECH) with acrylic acid (AA) and methacrylic acid (MAA) respectively. These monomers were synthesized via two different routes i.e. direct reaction of AA and MAA with ECH and second by reaction of AA and MAA with Sodium hydroxide to form sodium salt of the acid followed by reacting with ECH. The polymerization inhibitor used was tert-butyl hydroquinone (TBHQ) and catalysts used were triethylamine (TEA) and quaternary ammonium salt. Experimental results show that first route is suitable for synthesizing GA and second route is suitable for synthesis of GMA. The catalysts also have drastic effect on the conversion to the respective glycidyl esters. The study of the effect of catalyst and polymerization inhibitor has been carried out via both routes.

GBT021601, a Next Generation HbS Polymerization Inhibitor: Results of Safety, Tolerability, Pharmacokinetics and Pharmacodynamics in Adults Living with Sickle Cell Disease and Healthy Volunteers

Background: Sickle cell disease (SCD) is caused by polymerization of sickle hemoglobin (HbS), resulting in red blood cell (RBC) sickling, RBC destruction, vaso-occlusion and end-organ damage. GBT021601 is an oral, small molecule, next-generation HbS polymerization inhibitor. Similar to voxelotor, the first generation HbS polymerization inhibitor, GBT021601 increases hemoglobin-oxygen (Hb-O2) affinity and stabilizes hemoglobin (Hb) in the oxy-hemoglobin (oxyHb) state, thereby inhibiting polymerization of HbS in RBCs. The fraction of Hb bound to drug - (Hb occupancy) approximates the oxyHb molecules per RBC. Compared to voxelotor, GBT021601 has the potential to achieve higher Hb occupancies. GBT021601 achieves greater exposures per dose and is more potent as measured by improvements in hematological parameters in an in vivo SCD mouse model (Dufu, Kobina 2020). We hypothesized that GBT021601 would achieve a substantial reduction in RBC hemolysis and increase in hemoglobin while maintaining a favorable safety profile. We therefore explored safety, tolerability, pharmacokinetics (PK) and pharmacodynamics (PD) in healthy volunteer participants (HVs) and adults living with SCD. Methods: Two studies are in progress: 1) the first in human (FIH), randomized, double-blind, placebo-controlled, parallel group phase 1 study for HVs ages 18-55 years; and 2) a single arm, intrapatient single dose and multiple ascending dose study in homozygous HbSS SCD patients ages 18-60 years with baseline Hb levels ≥5.5 g/dL and ≤10.5 g/dL and without vaso-occlusive crisis or transfusion within 30 days of screening. The HV study tests single ascending doses (SAD) with a 6:2 randomization ratio. Doses administered to HV cohorts ranged from 50 mg to 2200 mg. The SCD patient study tests a single dose (100 mg) followed by multiple ascending doses of the study drug in the same patients to reach a % Hb occupancy of >20-30% over 7 weeks. The primary endpoints for both studies are safety and tolerability. Secondary endpoints include PK and PD. For the patient study, additional secondary endpoints are confirmation of the relationship between time matched GBT021601 concentrations and the change from baseline of clinical measures of anemia and hemolysis. Hb occupancy is calculated as a percentage of the molar ratio of drug concentrations in RBCs to the estimated Hb concentrations (using MCHC) in RBCs. Results: As of July 22, 2021, 63 HVs and 6 adults with SCD were enrolled. Thirty-nine HVs completed the study; one discontinued due to moving to a new geographical area and 23 were in follow-up. GBT021601 was generally well-tolerated, most adverse events (AE) were Grade 1 and 2, there were no deaths, and there was one serious adverse event in a HV that was not related to the study drug. There were no AEs indicative of tissue hypoxia. GBT021601 showed linear PK, high partitioning into RBCs, and a dose dependent increase in percent hemoglobin occupancy in HVs (Figure 1). From single doses of 50 to 1400 mg, the mean preliminary % Hb occupancy ranged from 0.88 to 25%, respectively, exceeding the Hb occupancies reported for HV receiving single doses of voxelotor over a similar range (Putz, 2017). Six adults with SCD have received a single 100 mg dose of GBT021601 and the dose has been well tolerated. Multiple-dose data in adults will be presented. Conclusions: Single doses of GBT021601 are well tolerated in HV and adults with SCD. Given its drug exposure, GBT021601 has the capacity to achieve a targeted Hb occupancy and attain the desired hematological effect at low doses, therefore reducing pill burden and improving clinical outcomes for individuals living with SCD. Forthcoming multiple-dose data will help to evaluate GBT021601's potential as a best-in-class, oral, disease-modifying therapy for SCD. Funding: This study was supported by Global Blood Therapeutics. Figure 1 Figure 1. Disclosures Brown: Pfizer: Research Funding; Imara: Consultancy, Research Funding; Novartis: Consultancy, Research Funding; Forma Therapeutics: Research Funding; Novo Nordisk: Consultancy; Global Blood Therapeutics: Consultancy, Research Funding. Redfern: Eisai: Other: Advisory Board; Linear Clinical Research: Current Employment; Pfizer: Other: Advisory Board; Novartis: Other: Advisory Board; Astra Zeneca: Other: Advisory Board; Roche: Other: Advisory Board. Lisbon: Global Blood Therapeutics: Current Employment. Washington: Global Blood Therapeutics: Consultancy. Agodoa: Global Blood Therapeutics: Current Employment, Current equity holder in publicly-traded company. Smith-Whitley: Global Blood Therapeutics: Current Employment.

Applications of the Poly(methyl methacrilate) (PMMA) in Cranioplasty

Cranioplasty is a surgical procedure that is used to correct any defect in skull bones after a previous decompresive craniectomy, usually made for traumatic brain injuries, with ischemic or hemorragic nature, or after a tumor removal. A composite for modeling on a defect in skull bones was prepared by crosslinking poly(methyl methacrylate) in the presence of barium sulfate. The crosslinking rate of methyl polymethacrylate in the presence of the benzoyl peroxide initiator, the N, N-dimethyl-β-toluidine polymerization accelerator and the hydroquinone polymerization inhibitor allows modeling according to the location and size of the defect. Thus, the concentration of composite precursor components was optimal for this purpose. The TGA diagram shows the almost total consumption of methyl methacrylate and butyl methacrylate monomers in the crosslinking process of methyl polymethacrylate with the formation of the composite. This technical study demonstrate the efficacy of this treatment, as well as to show all the possible scenarios in such procedures.

ВЛИЯНИЕ ЦЕЛОСТНОСТИ ЭЛЕМЕНТОВ ЦИТОСКЕЛЕТА И КАЛЬЦИЯ ВНУТРИКЛЕТОЧНЫХ ДЕПО НА ФУНКЦИОНАЛЬНЫЙ СТАТУС РАЗМОРОЖЕННЫХ СПЕРМАТОЗОИДОВ

Криоконсервация спермы — важный инструмент репродуктивных технологий, однако качество замороженной спермы остается невысоким. Предположено, что предварительная инкубация сперматозоидов быков до замораживания в присутствии определенных соединений может способствовать улучшению ее качества после разморозки. Предварительную инкубацию проводили с применением гепарина, который стимулирует в сперматозоидах быков увеличение количества капацитированных клеток. Другие используемые соединения — пролактин (ПРЛ) и гуанозинтрифосфат (ГТФ), при инкубации не увеличивали число капацитированных сперматозоидов. Добавление гепарина стимулировало освобождение Са2+ из внутриклеточных депо сперматозоидов быков, содержание которого снижалось в присутствии ингибитора полимеризации микрофиламентов цитохалазина Д, тогда как ингибитор полимеризации микротрубочек нокодазол не влиял на этот процесс. Активированное совместным действием ПРЛ и ГТФ освобождение Са2+ из внутриклеточных депо сперматозоидов отменялось в присутствии нокодазола, в то время как цитохалазин Д не влиял на этот процесс. После разморозки сперматозоидов увеличивается число капацитированных клеток, и чем их больше, тем хуже образец. С помощью хлортетрациклинового теста обнаружено, что после предварительной капацитации в присутствии гепарина, последующего замораживания и размораживания отмечалось снижение количества капацитированных сперматозоидов, в то же время использование для предварительной инкубации ПРЛ и ГТФ не приводило к уменьшению числа капацитированных клеток в размороженной сперме. Таким образом, предварительная инкубация сперматозоидов быков перед замораживанием в присутствии гепарина, действие которого связано с микрофиламентами, приводила к снижению числа капацитированных клеток после размораживания, что может свидетельствовать о возможном улучшении качества спермы после размораживания. INFLUENCE OF INTEGRITY OF CELLSKELETON AND CALCIUM ELEMENTS OF EXTRACELLULAR STORES ON THE FUNCTIONAL STATUS OF DEFROSTED SPERMATOZOA Sperm cryopreservation is an important tool of reproductive technologies, but the quality of frozen sperm remains low. It has been suggested that pre-incubation of bovine spermatozoa before freezing in the presence of certain compounds may provide to improving the quality of sperm after thawing. Pre-incubation was carried out with using of heparin, which stimulate an increase in the number of capacitated spermatozoa during incubation. Other compounds, which were used — prolactin (PRL) and guanosine triphosphate (GTP), did not increase the number of capacitated cells during incubation. The supplementation of heparin stimulated Ca2+ release from intracellular stores of bull spermatozoa, which was inhibited by microfilament polymerization inhibitor —cytochalasin D, while microtubule polymerization inhibitor nocodazole did not affect this process. The release of Ca2+ from intracellular sperm stores activated by complex action of PRL and GTP was canceled by nocodazole, while cytochalasin D did not affect this process. After thawing of sperm, the number of capacitated cells increases, and the higher their number is, the worse the sample is. By using chlortetracycline probe it was found that after preliminary capacitation conducted by heparin, subsequent freezing and thawing, the decrease in the number of capacitated cells was observed at the same time the usage of PRL for pre-incubation and GTP did not lead to the drop in the number of capacitated cells among thawed bovine spermatozoa. Thus, pre-incubation of bovine sperm before freezing in the presence of heparin, whose action is associated with microfilaments, led to a decrease in the number of capacitated cells after thawing that may indicate a possible improvement of sperm quality after thawing.

4’-O-Methylbroussochalcone B as a novel tubulin polymerization inhibitor suppressed the proliferation and migration of acute myeloid leukaemia cells

Background Recent years, survival rates of human with high-risk acute myeloid leukaemia (AML) have not raised substantially. This research aimed to investigate the role of 4′-O-Methylbroussochalcone B, for the treatment of human AML. Methods Firstly, we evaluated the effects of six chalcones on AML cells activity by MTT assay. Immunofluorescence staining, tubulin polymerization assay and N,N′-ethylenebis (iodoacetamide) (EBI) competition assay were performed on ML-2 cells. Transwell and apoptosis assay were also utilized in ML-2 cells and OCI-AML5 cells. The expressions of migration-related proteins, apoptosis-related proteins and Wnt/β-catenin pathway were detected by Western Blot. Results The results found six chalcones exhibited the anti-proliferative activity against different AML cell lines. Based on the results of immunofluorescence staining, tubulin polymerization assay and EBI competition assay, 4′-O-Methylbroussochalcone B was discovered to be a novel colchicine site tubulin polymerization inhibitor. 4′-O-Methylbroussochalcone B could induce apoptosis, inhibit proliferation and migration of ML-2 cells and OCI-AML5 cells. The cells were arrested in the G2-M phase by the treatment of 4′-O-Methylbroussochalcone B. In addition, 4′-O-Methylbroussochalcone B regulated MAPK and Wnt/β-catenin pathways in AML cells. Conclusion 4′-O-Methylbroussochalcone B might inhibit proliferation and migration of the AML cells by MAPK and Wnt/β-catenin pathways as a tubulin polymerization inhibitor. It is promising for 4′-O-Methylbroussochalcone B to become a new drug to treat AML.

Pyrinap ligands for enantioselective syntheses of amines

Amines are a class of compounds of essential importance in organic synthesis, pharmaceuticals and agrochemicals. Due to the importance of chirality in many practical applications of amines, enantioselective syntheses of amines are of high current interest. Here, we wish to report the development of (R,Ra)-N-Nap-Pyrinap and (R,Sa)-N-Nap-Pyrinap ligands working with CuBr to catalyze the enantioselective A3-coupling of terminal alkynes, aldehydes, and amines affording optically active propargylic amines, which are platform molecules for the effective derivatization to different chiral amines. With a catalyst loading as low as 0.1 mol% even in gram scale reactions, this protocol is applied to the late stage modification of some drug molecules with highly sensitive functionalities and the asymmetric synthesis of the tubulin polymerization inhibitor (S)-(-)-N-acetylcolchinol in four steps. Mechanistic studies reveal that, unlike reported catalysts, a monomeric copper(I) complex bearing a single chiral ligand is involved in the enantioselectivity-determining step.