Microwave-assisted synthesis of a MK2 inhibitor by Suzuki-Miyaura coupling for study in Werner syndrome cells

Microwave-assisted Suzuki-Miyaura cross-coupling reactions have been employed towards the synthesis of three different MAPKAPK2 (MK2) inhibitors to study accelerated aging in Werner syndrome (WS) cells, including the cross-coupling of a 2-chloroquinoline with a 3-pyridinylboronic acid, the coupling of an aryl bromide with an indolylboronic acid and the reaction of a 3-amino-4-bromopyrazole with 4-carbamoylphenylboronic acid. In all of these processes, the Suzuki-Miyaura reaction was fast and relatively efficient using a palladium catalyst under microwave irradiation. The process was incorporated into a rapid 3-step microwave-assisted method for the synthesis of a MK2 inhibitor involving 3-aminopyrazole formation, pyrazole C-4 bromination using N-bromosuccinimide (NBS), and Suzuki-Miyaura cross-coupling of the pyrazolyl bromide with 4-carbamoylphenylboronic acid to give the target 4-arylpyrazole in 35% overall yield, suitable for study in WS cells

Age-dependent alterations in spermatogenesis in itchy mice

Spermatogenesis is an intricate process in which spermatogonial stem cells divide and differentiate to produce mature sperm. This process strongly depends on protein turnover both in the developing germ cells and the supportive Sertoli cells, and recent evidence has demonstrated the role of the ubiquitin-proteasome system in this protein turnover in the testis. Itch, an E3 ligase important in the immune system, has been implicated in regulating the blood testis barrier. Although the specific role of Itch during spermatogenesis is not yet well understood, its ubiquitous expression and wide array of functional targets suggest multiple and tissue-specific roles. Here the testes of mice that lack Itch protein are evaluated at two developmental time points: peri-pubertal postnatal day (PND) 28 and adult PND 56. Itchy mice demonstrate an increased germ cell apoptotic index compared with wild type C57BL/6J mice at both PND 28 and PND 56. A corresponding 27% reduction in the total number of spermatid heads produced in PND 56 itchy mice was also evident. A histological evaluation of itchy mice revealed a delay in spermatogenesis at PND 28 and disorganization of late stage spermatids at PND 56. An analysis of several apoptotic markers revealed an age-dependent change in cleaved caspase 9, an intrinsic apoptosis mediator. The breeding success of the itchy mice was also significantly decreased, possibly due to a developmental defect. Taken together, these findings indicate that Itch is required for functional spermatogenesis, and that it may play differing cellular roles during development

Microwave-assisted synthesis of a MK2 inhibitor by Suzuki-Miyaura coupling for study in Werner syndrome cells

Microwave-assisted Suzuki-Miyaura cross-coupling reactions have been employed towards the synthesis of three different MAPKAPK2 (MK2) inhibitors to study accelerated aging in Werner syndrome (WS) cells, including the cross-coupling of a 2-chloroquinoline with a 3-pyridinylboronic acid, the coupling of an aryl bromide with an indolylboronic acid and the reaction of a 3-amino-4-bromopyrazole with 4-carbamoylphenylboronic acid. In all of these processes, the Suzuki-Miyaura reaction was fast and relatively efficient using a palladium catalyst under microwave irradiation. The process was incorporated into a rapid 3-step microwave-assisted method for the synthesis of a MK2 inhibitor involving 3-aminopyrazole formation, pyrazole C-4 bromination using N-bromosuccinimide (NBS), and Suzuki-Miyaura cross-coupling of the pyrazolyl bromide with 4-carbamoylphenylboronic acid to give the target 4-arylpyrazole in 35% overall yield, suitable for study in WS cells

Building molecules for the modulation of age-related diseases

Improved understanding of normal human ageing will provide important insights into major risk factors for many age-related diseases. Using the progeroid disease Werner syndrome (WS) as a model for accelerated ageing, this project aimed to investigate how modulation of protein function in the p38α stress-signalling kinase cascade could affect ageing on the cellular level. New rapid routes towards p38α and MK2 inhibitors have been developed to evaluate their use as chemical probes to investigate the mechanisms of cellular ageing. The p38α inhibitor RO3201195 has been synthesized and used as a probe to corroborate previously published evidence that inhibition of the p38α protein kinase reverses the aged morphology of WS fibroblasts. A new route for the synthesis of the MK2 inhibitor PF-3644022 was developed, exploring new methods for building the heterocyclic scaffold through a number of retrosynthetic strategies. Microwave-assisted organic synthesis was used extensively as a tool towards halogenated benzothiophenes and fused quinoline systems, and in rapid Suzuki-Miyaura coupling and Buchwald-Hartwig N-arylation chemistry. This MK2 inhibitor can now be used as a readily accessible probe to further investigate the role of kinases downstream of p38α and the role of MK2 in the premature ageing of WS cells. The final part of the project focussed on the potential of a known p38α inhibitor, BIRB 796, in the development of new chemical probes. BIRB 796 binds allosterically outside of the ATPbinding pocket of p38α MAPK, which drives a conformational change of the kinase into an inactive form. Using rapid synthetic methods towards pyrazole and urea formation, the selectivity elements of the inhibitor were reviewed and a new library of analogues was synthesized. Upon testing against a small panel of kinases, it was found that through small changes the inhibitor potency for p38α could be dramatically altered, and binding affinity for alternative kinases could be selectively enhanced

Microwave-assisted synthesis of 3-aminobenzo[b]thiophene scaffolds for the preparation of kinase inhibitors

Microwave irradiation of 2-halobenzonitriles and methyl thioglycolate in the presence of triethylamine in DMSO at 130 °C provides rapid access to 3-aminobenzo[b]thiophenes in 58–96% yield. This transformation has been applied in the synthesis of the thieno[2,3-b]pyridine core motif of LIMK1 inhibitors, the benzo[4,5]thieno[3,2-e][1,4]diazepin-5(2H)-one scaffold of MK2 inhibitors and a benzo[4,5]thieno[3,2-d]pyrimidin-4-one inhibitor of the PIM kinases

The effect of RO3201195 and a pyrazolyl ketone P38 MAPK inhibitor library on the proliferation of Werner syndrome cells

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Annotated Bibliography: Literature Review Of Causative And Non-Causative Risk Factors For Breast Cancer-Related Lymphedema

Annotated bibliography derived from a review of literature concerning causative and non-causative risk factors for breast cancer-related lymphedema. This document was created by UNE Doctor of Physical Therapy students to establish a knowledge base for clinicians to better understand, diagnose, and educate patients.https://dune.une.edu/lrr/1000/thumbnail.jp