Discovery of the Potent and Selective ATR Inhibitor Camonsertib (RP-3500).

ATR is a key kinase in the DNA-damage response (DDR) that is synthetic lethal with several other DDR proteins, making it an attractive target for the treatment of genetically selected solid tumors. Herein we describe the discovery of a novel ATR inhibitor guided by a pharmacophore model to position a key hydrogen bond. Optimization was driven by potency and selectivity over the related kinase mTOR, resulting in the identification of camonsertib (RP-3500) with high potency and excellent ADME properties. Preclinical evaluation focused on the impact of camonsertib on myelosuppression, and an exploration of intermittent dosing schedules to allow recovery of the erythroid compartment and mitigate anemia. Camonsertib is currently undergoing clinical evaluation both as a single agent and in combination with talazoparib, olaparib, niraparib, lunresertib, or gemcitabine (NCT04497116, NCT04972110, NCT04855656). A preliminary recommended phase 2 dose for monotherapy was identified as 160 mg QD given 3 days/week.

A cadaveric assessment of percutaneous trigger finger release with 15° stab knife: its effectiveness and complications.

Percutaneous release of the A1 pulley has been introduced as a therapeutic approach for trigger fingers and is suggested as an effective and safe alternative, where conservative treatments fail. The aim of the current study was to determine if percutaneous release with a 15° stab knife can effectively result in acceptable efficacy and lower complication rate. METHODS: In the present study, the percutaneous release of the A1 pulley was evaluated by percutaneous release using a 15° stab knife in 20 fresh-frozen cadaver hands (10 cadavers). One hundred fingers were finally included in the present study. The success rate of A1 pulley release as well as the complications of this method including digital vascular injury, A2 pulley injury, and superficial flexor tendon injury was evaluated, and finally, the data were analyzed by the SPSS software. RESULTS: The results showed a success rate of 75% for A1 pulley release in four fingers, followed by eleven fingers (90%) and eighty-five fingers (100%). Therefore, the A1 pulley was found to be completely released in eighty-five fingers (100%). Overall, the mean of A1 pulley release for these fingers was determined as 97.9%, indicating that percutaneous trigger finger release can be an effective technique using a 15° stab knife. Furthermore, our findings revealed no significant difference in the amount of A1 pulley release in each of the fingers in the right and left hands. Additionally, 17 fingers developed superficial scrape in flexor tendons, while 83 fingers showed no flexor tendons injuries and no other injuries (i.e., vascular, digital nerve, and A2 pulley injuries). CONCLUSIONS: Percutaneous release of the A1 pulley using a 15° stab knife was contributed to acceptable efficacy and a relatively good safety in the cadaveric model.

Comparing the Corticosteroid Injection and A1 Pulley Percutaneous Release in Treatment of Trigger Finger: A Clinical Trial.

Background: Primary treatment of trigger digits is conservative including stretching, night splinting and combination of heat and ice. When these methods fail, invasive methods such as corticosteroid injection, percutaneous release and open surgery are used. The purpose of this study is to compare the efficacy of two outpatient methods of percutaneous trigger finger release (PTFR) and corticosteroid injection (CI). Methods: This study is a randomized clinical trial that was performed with 6-month follow up. A total of 83 patients with trigger finger treated either with corticosteroid injection (n:40) or percutaneous release of the A1 pulley (n:43) were enrolled in this study. Demographic data were recorded before intervention. Pain score (VAS criterion), disease stage (Quinnell criteria), patient satisfaction and complications such as paresthesia, scarring, and stiffness (decrease in the range of motion) were recorded after the intervention. We used SPSS program (statistical package for the social science SPSS version 16) to perform the analysis. Results: There were 18 male (21.7%) and 65 female (78.3%) patients, whose mean age was 52.54 ± 11.45 (28-85) years. There was a significant difference between the degree of pain at the time of the third, sixth weeks and sixth months in two groups. The degree of pain was lower in the CI group in the third and sixth weeks but it was lower in the PTFR group in the sixth month. Satisfaction of the patients in the sixth month was significantly higher in the PTFR group. The incidence of stiffness was also significantly lower in the PTFR group in the sixth month. Conclusions: Patients in PTFR group had greater recovery and satisfaction level and lower recurrence rate and pain. Therefore PTFR may be used as a substitute for CI in the treatment of trigger finger from the beginning especially in patients who do not want to have open surgery.

Treatment of Distal Humerus Articular Fracture with Pin-and-Plate Technique.

Background: Distal Humerus Articular Fracture (classified by Arbeitsgemeinschaft für Osteosynthesefragen/Orthopedic Trauma Association as 13-B3) is an uncommon fracture with significant complications. We report results of 14 patients treated by open reduction and pin-and-plate fixation technique. Methods: In this retrospective study, we applied pin-and-plate fixation to 14 distal humerus articular fractures, in which screw fixation alone was inefficient or inadequate to provide stable fixation. After anatomical reduction of articular fragments, multiple Kirschner wires were inserted through the fragments. To prevent migration of Kirschner wires a small plate was placed proximally on the bent end of the pins. Results: The average age of 14 patients (8 males and 6 females) was 36.4 years (range: 16-57) and the mean follow up period was 43 months (range: 12-80). At last follow up the average quick Disabilities of the Arm, Shoulder and Hand score was 18.9 (range: 2.3-42.5) and the mean points for Mayo Elbow Performance Index was 75.3 (range: 50-100). Mean final arc of flexion-extension was 97° (range: 40-131). Conclusions: Distal humerus articular fracture is sometimes difficult to fix with conventional methods. We used pin-and-plate technique that could make a stable fixation and allow early range of motion with acceptable results.

Ecological Roles of Tryptanthrin, Indirubin and N-Formylanthranilic Acid in Isatis indigotica: Phytoalexins or Phytoanticipins?

Leaves of the plant species Isatis indigotica Fortune ex Lindl. (Chinese woad) produce the metabolites tryptanthrin, indirubin and N-formylanthranilic acid upon spraying with an aqueous solution of copper chloride but not after spraying with water. The antifungal activities of these metabolites against the phytopathogens Alternaria brassicicola, Leptosphaeria maculans and Sclerotinia sclerotiorum established that tryptanthrin is a much stronger growth inhibitor of L. maculans than the phytoalexin camalexin. The biosynthetic precursors of tryptanthrin and N-formylanthranilic acid are proposed based on the deuterium incorporations of isotopically labeled compounds. The overall results suggest that tryptanthrin is a phytoalexin and indirubin and N-formylanthranilic acid are phytoanticipins in the plant species I. indigotica and that chemical diversity and biodiversity are intimately connected.

Methoxycamalexins and related compounds: Syntheses, antifungal activity and inhibition of brassinin oxidase.

The phytoalexin camalexin is a competitive inhibitor of brassinin oxidase, an enzyme that detoxifies the phytoalexin brassinin and is produced by an economically important plant pathogen. For this reason, the camalexin scaffold has guided the design of inhibitors of brassinin detoxification. To further understand the structure-activity relationships of camalexin related compounds, the syntheses of monomethoxy and dimethoxycamalexins were undertaken. Four monomethoxy camalexins together with 4,6-dimethoxy and 5,7-dimethoxy camalexins were prepared from the corresponding methoxyindoles using the Ayer's method. The dimethoxy derivatives were prepared from the corresponding dimethoxyindole-3-thiocarboxamides using the Hantzsch reaction; however, this method did not work for the syntheses of 4,6-dimethoxy and 5,7-dimethoxycamalexins due to the lower reactivities of the corresponding indole-3-thiocarboxamides. The antifungal activity and brassinin oxidase inhibitory activity of all methoxycamalexins and ten camalexin related compounds were investigated. Among the 20 compounds evaluated, monomethoxycamalexins were stronger antifungals than the dimethoxy derivatives. However, remarkably, 5,6-dimethoxycamalexin, 6,7-dimethoxycamalexin and 5-methoxycamalexin displayed the strongest inhibitory activity against brassinin oxidase, while 4,5-dimethoxycamalexin displayed no inhibitory effect. Altogether the structure-activity relationships of camalexin related compounds suggest that the targets for fungal growth inhibition and brassinin oxidase inhibition are unrelated and emphasize that brassinin oxidase inhibitors do not need to be antifungal.

Inhibitors of the Detoxifying Enzyme of the Phytoalexin Brassinin Based on Quinoline and Isoquinoline Scaffolds.

The detoxification of the phytoalexin brassinin to indole-3-carboxaldehyde and S-methyl dithiocarbamate is catalyzed by brassinin oxidase (BOLm), an inducible fungal enzyme produced by the plant pathogen Leptosphaeria maculans. Twenty-six substituted quinolines and isoquinolines are synthesized and evaluated for antifungal activity against L. maculans and inhibition of BOLm. Eleven compounds that inhibit BOLm activity are reported, of which 3-ethyl-6-phenylquinoline displays the highest inhibitory effect. In general, substituted 3-phenylquinolines show significantly higher inhibitory activities than the corresponding 2-phenylquinolines. Overall, these results indicate that the quinoline scaffold is a good lead to design paldoxins (phytoalexin detoxification inhibitors) that inhibit the detoxification of brassinin by L. maculans.

Biotransformation of rutabaga phytoalexins by the fungus Alternaria brassicicola: Unveiling the first hybrid metabolite derived from a phytoalexin and a fungal polyketide.

The biotransformations of the rutabaga phytoalexins rutalexin, brassicanate A, isalexin and rapalexin A by the plant pathogenic fungus Alternaria brassicicola are reported. While the biotransformations of rutalexin, brassicanate A, and isalexin are fast, rapalexin A is resistant to fungal transformation. Unexpectedly, biotransformation of rutalexin yields a hybrid metabolite named rutapyrone, derived from rutalexin metabolism and phomapyrone G, a fungal metabolite produced by A. brassicicola. These fungal transformations are detoxification reactions likely carried out by different enzymes. The discovery of rapalexin A resistance to detoxification suggests that this phytoalexin in combination with additional phytoalexins could protect crucifers against this pathogen. Phytoalexins resistant to degradation by A. brassicicola are expected to provide the producing plants with higher disease resistance levels.

Evaluation of differences between two groups of low back pain patients with and without rotational demand activities based on hip and lumbopelvic movement patterns.

BACKGROUND: Excessive and earlier lumbopelvic motions during trunk and limb movements tests have been reported in both low back pain (LBP) patients with and without trunk and hip rotational demand activities. The aim of the present study was to determine differences in hip and lumbopelvic rotation during the active hip internal rotation (AHIR) test between two groups of LBP patients with and without regular trunk and hip rotational demand activities. MATERIAL AND METHODS: A total of 35 LBP patients, including 15 males who regularly participated in rotational demand sports activities and 20 males not participating in sports and functional rotational demand activities, participated in study. The AHIR test was performed. The kinematic variables of hip and pelvic rotations were recorded by a Qualisys motion analysis system. Pelvic and hip rotations were calculated across time during the test. In addition, pelvic rotations in the first half of the test and pelvic-hip timing were calculated. RESULTS: People with rotational demand activities had a higher pelvic rotation both during the test and in the first 50% of movement. Earlier pelvic rotation was observed in people with rotational demand activities compared to people with non-rotational demand activities. CONCLUSION: 1. The data of the current study suggests that lumbopelvic movement patterns in different groups of LBP patients in regard to their specific activities may vary. 2. LBP people with rotational demand sports activities have a greater tendency of pelvic rotation motion during the AHIR.

The phytoalexin camalexin induces fundamental changes in the proteome of Alternaria brassicicola different from those caused by brassinin.

Camalexin is the major phytoalexin produced by Alternaria thaliana, but is absent in Brassica species that usually produce phytoalexin blends containing brassinin and derivatives. The protein profiles of A. brassicicola treated with camalexin were evaluated using proteomics and metabolic analyses and compared with those treated with brassinin. Conidial germination and mycelial growth of A. brassicicola in liquid media amended with camalexin and brassinin showed that fungal growth was substantially slower in presence of camalexin than brassinin; chemical analyses revealed that A. brassicicola detoxified camalexin at much slower rate than brassinin. Two-dimensional gel electrophoresis (2-DE) followed by tryptic digestion and capillary liquid chromatography-mass spectrometric analyses identified 158 different proteins, of which 45 were up-regulated and 113 were down-regulated relative to controls. Venn diagram analyses of differentially expressed proteins in cultures of A. brassicicola incubated with camalexin and brassinin indicated clear differences in the effect of each phytoalexin, with camalexin causing down-regulation of a larger number of proteins than brassinin. Overall, results of this work suggest that each phytoalexin has several different targets in the cells of A. brassicicola, and that camalexin appears to have greater potential to protect cultivated Brassica species against A. brassicicola than brassinin.

Metabolism of the phytoalexins camalexins, their bioisosteres and analogues in the plant pathogenic fungus Alternaria brassicicola.

The metabolism of the phytoalexins camalexin (1), 1-methylcamalexin (10) and 6-methoxycamalexin (11) by Alternaria brassicicola and their antifungal activity is reported. This work establishes that camalexins are slowly biotransformed (ca. six days) to the corresponding indole-3-thiocarboxamides, which are further transformed to the indole-3-carboxylic acids. These metabolites are substantially less inhibitory to A. brassicicola than the parent camalexins, indicating that these enzyme-mediated transformations are detoxifications. In addition, analyses of the metabolism of synthetic isomers and bioisosteres of camalexin (1) indicate that isomers of camalexin in the thiazole ring are not metabolized. Based on these results, the potential intermediates that lead to formation of indole-3-thiocarboxamides are proposed.

Unprecedented spirocyclization of 3-methyleneindoline-2-thiones during hydrolysis of the phytoalexin cyclobrassinin.

The phytoalexin cyclobrassinin is a plant defense that has additional importance since it inhibits brassinin hydrolase, a phytoalexin detoxifying enzyme produced by the plant pathogen Alternaria brassicicola. Hence, the 1,3-thiazino[6,5-b]indole scaffold of cyclobrassinin has great application as a lead structure to design potential inhibitors of brassinin detoxification. For this reason, it is necessary to determine whether A. brassicicola is able to transform cyclobrassinin. During this work new reactions of 1,3-thiazino[6,5-b]indoles and indoline-2-thiones and their unique [4+2] cycloaddition products were discovered and characterized.

Synthesis of new tripodal Hantzsch 1,4-dihydropyridines under solvent-free condition and their conversion to the corresponding tripodal pyridines.

A mixture of ß-ketoester and tri-aldehydes in the presence of ammonium fluoride was converted to their corresponding tripodal 1,4-dihydropyridines under solvent-free condition with good yields. The obtained tripodal 1,4-dihydropyridines were also aromatized with oxone(®)/NaNO(2)/wet SiO(2) system under mild and heterogeneous conditions quantitatively.

A simple and efficient route for the synthesis of di and tri(bis(indolyl) methanes) as new triarylmethanes.

Preparation of di and tri(bis(indolyl) methanes) from di and trialdehydes and indoles in the presence of silica sulfuric acid was described. Reactions proceeded in good to excellent yields in acetonitrile as solvent at room temperature.