Synthesis and Preliminary Characterization of Putative Anle138b-Centered PROTACs against α-Synuclein Aggregation.

The search for disease-modifying agents targeted against Parkinson's disease led us to rationally design a small array of six Anle138b-centered PROTACs, 7a,b, 8a,b and 9a,b, targeting αSynuclein (αSyn) aggregates for binding, polyubiquitination by the E3 ligase Cereblon (CRBN), and proteasomal degradation. Lenalidomide and thalidomide were used as CRBN ligands and coupled with amino- and azido Anle138b derivatives through flexible linkers and coupling reactions (amidation, 'click' chemistry). Four Anle138b-PROTACs, 8a,b and 9a,b, were characterized against in vitro αSyn aggregation, monitoring them in a Thioflavin T (ThT) fluorescence assay and in dopaminergic neurons derived from a set of isogenic pluripotent stem cell (iPSC) lines with SNCA multiplications. Native and seeded αSyn aggregation was determined with a new biosensor, and a partial correlation between αSyn aggregation, cellular dysfunctions, and neuronal survival was obtained. Anle138b-PROTAC 8a was characterized as the most promising αSyn aggregation inhibitor/degradation inducer, with potential usefulness against synucleinopathies and cancer.

Synthesis of SARS-CoV-2 Mpro inhibitors bearing a cinnamic ester warhead with in vitro activity against human coronaviruses.

COVID-19 now ranks among the most devastating global pandemics in history. The causative virus, SARS-CoV-2, is a new human coronavirus (hCoV) that spreads among humans and animals. Great efforts have been made to develop therapeutic agents to treat COVID-19, and among the available viral molecular targets, the cysteine protease SARS-CoV-2 Mpro is considered the most appealing one due to its essential role in viral replication. However, the inhibition of Mpro activity is an interesting challenge and several small molecules and peptidomimetics have been synthesized for this purpose. In this work, the Michael acceptor cinnamic ester was employed as an electrophilic warhead for the covalent inhibition of Mpro by endowing some peptidomimetic derivatives with such a functionality. Among the synthesized compounds, the indole-based inhibitors 17 and 18 efficiently impaired the in vitro replication of beta hCoV-OC-43 in the low micromolar range (EC50 = 9.14 µM and 10.1 µM, respectively). Moreover, the carbamate derivative 12 showed an antiviral activity of note (EC50 = 5.27 µM) against another hCoV, namely hCoV-229E, thus suggesting the potential applicability of such cinnamic pseudopeptides also against human alpha CoVs. Taken together, these results support the feasibility of considering the cinnamic framework for the development of new Mpro inhibitors endowed with antiviral activity against human coronaviruses.

Discovery of a Novel Trifluoromethyl Diazirine Inhibitor of SARS-CoV-2 Mpro.

SARS-CoV-2 Mpro is a chymotrypsin-like cysteine protease playing a relevant role during the replication and infectivity of SARS-CoV-2, the coronavirus responsible for COVID-19. The binding site of Mpro is characterized by the presence of a catalytic Cys145 which carries out the hydrolytic activity of the enzyme. As a consequence, several Mpro inhibitors have been proposed to date in order to fight the COVID-19 pandemic. In our work, we designed, synthesized and biologically evaluated MPD112, a novel inhibitor of SARS-CoV-2 Mpro bearing a trifluoromethyl diazirine moiety. MPD112 displayed in vitro inhibition activity against SARS-CoV-2 Mpro at a low micromolar level (IC50 = 4.1 µM) in a FRET-based assay. Moreover, an inhibition assay against PLpro revealed lack of inhibition, assuring the selectivity of the compound for the Mpro. Furthermore, the target compound MPD112 was docked within the binding site of the enzyme to predict the established intermolecular interactions in silico. MPD112 was subsequently tested on the HCT-8 cell line to evaluate its effect on human cells' viability, displaying good tolerability, demonstrating the promising biological compatibility and activity of a trifluoromethyl diazirine moiety in the design and development of SARS-CoV-2 Mpro binders.

Metacarpophalangeal joint hyperextension in rhizartrosis: is surgical correction necessary?

BACKGROUND AND AIM: Rhizarthrosis represents 10% of all arthritic manifestations and its prevalence increases with age and in women. The hyperextension of the metacarpophalangeal joint (MCPj) is consequent to a progressive dorsoradial subluxation of the trapeziometacarpal joint (TMj) in advanced osteoarthritis. The aim of this retrospective study is to evaluate the clinical and functional results of 32 patients affected by advanced rhizarthrosis who underwent to modified Burton-Pellegrini's trapeziectomy in absence of surgical correction of MCPj hyperextension in order to understand when this last step is really necessary. METHODS: Patients were assessed trough DASH and PRWHE questionnaires; the functionality of the hand was assessed by carrying out specific test (grip strength, key-pinch, kapandji test, reduction of wrist flexion strength) and the degree of MCP joint hyperextension was recorded. RESULTS: Clinical evaluation and individual satisfactory were positive in most cases (mean DASH 19 and mean PRWHE 21.8, with a reduction of 77% of VAS pain score). Kapandji test was excellent in 26 patients and grip strength and key pinch were stackable in operated and non-operated hands. Twenty-five out 32 patients presented a MCP joint hyperextension between 0° and 5°, 5 of 10° and other 2 of 15°. CONCLUSION: Modified Burton-Pellegrini's trapeziectomy is a valid option to treat patient with TMj osteoarthritis. The absence of surgical correction of the MCPj does not affect clinical and functional results in deformities <15°.

Interfering with the Tumor-Immune Interface: Making Way for Triazine-Based Small Molecules as Novel PD-L1 Inhibitors.

The inhibition of the PD-1/PD-L1 axis by monoclonal antibodies has achieved remarkable success in treating a growing number of cancers. However, a novel class of small organic molecules, with BMS-202 (1) as the lead, is emerging as direct PD-L1 inhibitors. Herein, we report a series of 2,4,6-tri- and 2,4-disubstituted 1,3,5-triazines, which were synthesized and assayed for their PD-L1 binding by NMR and homogeneous time-resolved fluorescence. Among them, compound 10 demonstrated to strongly bind with the PD-L1 protein and challenged it in a co-culture of PD-L1 expressing cancer cells (PC9 and HCC827 cells) and peripheral blood mononuclear cells enhanced antitumor immune activity of the latter. Compound 10 significantly increased interferon γ release and apoptotic induction of cancer cells, with low cytotoxicity in healthy cells when compared to 1, thus paving the way for subsequent preclinical optimization and medical applications.

CXCR4 antagonism sensitizes cancer cells to novel indole-based MDM2/4 inhibitors in glioblastoma multiforme.

Glioblastoma Multiforme (GBM) is a highly invasive primary brain tumour characterized by chemo- and radio-resistance and poor overall survival. GBM can present an aberrant functionality of p53, caused by the overexpression of the murine double minute 2 protein (MDM2) and its analogue MDM4, which may influence the response to conventional therapies. Moreover, tumour resistance/invasiveness has been recently attributed to an overexpression of the chemokine receptor CXCR4, identified as a pivotal mediator of glioma neovascularization. Notably, CXCR4 and MDM2-4 cooperate in promoting tumour invasion and progression. Although CXCR4 actively promotes MDM2 activation leading to p53 inactivation, MDM2-4 knockdown induces the downregulation of CXCR4 gene transcription. Our study aimed to assess if the CXCR4 signal blockade could enhance glioma cells' sensitivity to the inhibition of the p53-MDMs axis. Rationally designed inhibitors of MDM2/4 were combined with the CXCR4 antagonist, AMD3100, in human GBM cells and GBM stem-like cells (neurospheres), which are crucial for tumour recurrence and chemotherapy resistance. The dual MDM2/4 inhibitor RS3594 and the CXCR4 antagonist AMD3100 reduced GBM cell invasiveness and migration in single-agent treatment and mainly in combination. AMD3100 sensitized GBM cells to the antiproliferative activity of RS3594. It is noteworthy that these two compounds present synergic effects on cancer stem components: RS3594 inhibited the growth and formation of neurospheres, AMD3100 induced differentiation of neurospheres while enhancing RS3594 effectiveness preventing their proliferation/clonogenicity. These results confirm that blocking CXCR4/MDM2/4 represents a valuable strategy to reduce GBM proliferation and invasiveness, acting on the stem cell component too.

Subtrocantheric nonunion following fracture of an arthrodesed hip: a case report.

In recent years in the era of successful of total hip replacement, hip arthrodesis is rarely performed. The anatomy and biomechanics of an arthrodesed hip is altered, thus influencing the treatments strategies in case of fracture or nonunions. This case report describes the management and therapeutic solution for the treatment of subtrochanteric nonunion in a patient with hip arthrodesis. Satisfactory outcomes were finally obtained after a double surgical procedure.

Efficacy and safety of viscosupplementation with hyaluronic acid for hip osteoarthritis: results from a cross-sectional study with a minimum follow-up of 4 years.

BACKGROUND AND AIM OF THE WORK: Osteoarthritis is the most common cause of disability in elderly. Hip osteoarthritis is the second most frequent form affecting a large joint and the social and economic impact on society of its related disability is expected to increase. The purpose of this study was to verify the efficacy and safety of ultrasound-guided viscosupplementation with high weight hyaluronic acid in hip osteoarthritis. METHODS: 183 patients with painful hip OA (Kellgren-Lawrence 1-2-3) were treated from January 2014 to December 2016 with viscosupplementation. Patients were evaluated before injection (T0) and after 1,2,3,4 years through the VAS scale and Harris Hip Score (HHS). Patients who underwent to subsequent injections were followed and assessed. Subjects who underwent prosthesis were analyzed for a minimum of 6 months in order to detect any early postoperative complication. RESULTS: The mean improvement of HHS and VAS between T0 and T1 was statistically significant. Patients who underwent subsequent injections showed a higher improvement even if statistical significance was not observed. Results showed that patients with grade 2 of osteoarthritis had the higher change in the scores. No adverse effects were registered. No early complications were reported in those patients who needed prosthesis. DISCUSSION AND CONCLUSIONS: Results observed confirm that ultrasound-guided viscosupplementation with high weight hyaluronic acid could be a possibility in the treatment of hip osteoarthritis, especially in patients with Kellgren-Lawrence grade 2 of disease. Subsequent injections are not characterized by similar positive effects. Outcomes of prosthetic surgery are not influenced by viscosupplementation.

Risks of subsequent contralateral fractures of the trochanteric region in elderly.

BACKGROUND: Fractures in elderly are always a dramatic event and the healing is often not complete. In a context of bone fragility, repeated fractures are a growing problem in the industrialized world, in which the mean age of population is increasing. The aim of this study was to identify those general factors which may increase the risk of subsequent trochanteric fractures after an initial lesion. MATERIALS AND METHODS: Three-hundred and thirty-one patients who underwent intramedullary fixation for trochanteric fractures between January 2012 and December 2013 were studied. Forty subjects yet alive (group 1), affected by a subsequent contralateral hip fracture, were compared with 202 patients (group 2) affected by isolated trochanteric fracture. Days of hospitalization before surgery, hospitalization, period of rehabilitation, type of discharge and comorbidities, that are reported in literature as possible risk factors for hip refracture, were analyzed. In addition, all patients were interviewed in order to assess if a therapy for osteoporosis was prescribed after the initial fracture and how their gait had been modified by fractures. RESULTS: Days of hospitalization before surgery, hospitalization, period of rehabilitation and type of discharge were not predictive factors for subsequent fractures, as well as diabetes mellitus, hypertension and cardiac diseases. The presence of neurologic and respiratory diseases were associated to a higher risk of refractures, as well as the absence of specific medical treatment for osteoporosis. CONCLUSIONS: Neurologic and respiratory comorbidities and the absence of osteoporosis medical treatment are the variables associated to a higher risk of contralateral fractures. Physicians can do more in terms of prevention and strategies must consider these risk factors.

Neuromuscular taping versus sham therapy on muscular strength and motor performance in multiple sclerosis patients.

PURPOSE: Purpose of this study is to evaluate differences in leg muscles strength and motor performance between neuromuscular taping (NT) and sham tape groups. METHOD: Relapsing-remitting (RR) multiple sclerosis (MS) patients were recruited and randomly assigned to NT or sham tape groups. All patients underwent the treatment 5 times at 5-d intervals. They were submitted to a 6-minute walk test and isokinetic test (peak torque) at the beginning (T0), at the end (T1) and 2 months after the end of the treatment (T2). RESULTS: Forty MS patients (38 F; 2 M; mean age 45.5 ± 6.5 years) were assigned to NT group (n = 20) and to sham tape group (n = 20). Delta Peak Torque T1-T0 and T2-T0 between two groups were statistically significant in quadriceps (p = 0.007; 0.000) and hamstrings (p = 0.011; 0.007). The difference between the two groups according to 6-minute walk test was not statistically significant but in NT group it was noticed an increasing trend about the distance run. CONCLUSIONS: In this single-blind randomized controlled trial, NT seemed to increase strength in leg muscles, compared to a sham device, in RR MS patients. Further studies are needed to consider this therapy as a complement to classic physical therapy. IMPLICATIONS FOR REHABILITATION: Neuromuscular taping (NT) in multiple sclerosis: NT is well tolerated by multiple sclerosis patients and should be a complement to classic physical therapy. This technique normalizes muscular function, strengthens weakened muscles and assists the postural alignment.

Use of platelet-rich plasma in the care of sports injuries: our experience with ultrasound-guided injection.

BACKGROUND: Platelet-rich plasma is being used more frequently to promote healing of muscle injuries. The growth factors contained in platelet-rich plasma accelerate physiological healing processes and the use of these factors is simple and minimally invasive. The aim of this study was to demonstrate the efficacy of ultrasound-guided injection of platelet-rich plasma in muscle strains and the absence of side effects. MATERIALS AND METHODS: Fifty-three recreational athletes were enrolled in the study. The patients were recruited from the Emergency Room in the University Hospital at Parma according to a pre-defined protocol. Every patient was assessed by ultrasound imaging to evaluate the extent and degree of muscle injuries. Only grade II lesions were treated with three ultrasound-guided injections of autologous platelet-rich plasma every 7 days. Platelet concentrate was produced according to standard methods, with a 10% variability in platelet count. The platelet gel for clinical use was obtained by adding thrombin to the concentrates under standardised conditions. Outcomes assessed were: pain reduction, muscle function recovery and return to sports activity, ultrasound-imaging tissue healing, relapses, local infections, and any side effect during the treatment. RESULTS: In all cases muscle lesions healed fully on ultrasound-imaging, the pain disappeared, and muscle function recovery was documented with a return to sports activity. A single patient had a relapse 1 year after treatment. DISCUSSION: Platelet-rich plasma injected into the injury site is one of the most important factors rendering the treatment effective. To maximise its efficacy the preliminary ultrasound must be done accurately to localise the lesion and guide the needle into the corresponding lesion. According to the current results, which document full muscle recovery and no relapse except for one case, platelet-rich plasma ultrasound-guided injection represents a valid mini-invasive treatment for muscle injuries.