In silico and in vitro metabolism of ribociclib: a mass spectrometric approach to bioactivation pathway elucidation and metabolite profiling

Ribociclib (RBC, Kisqali®) is a highly selective CDK4/6 inhibitor that has been approved for breast cancer therapy. Initially, prediction of susceptible sites of metabolism and reactivity pathways were performed by the StarDrop WhichP450™ module and the Xenosite web predictor tool, respectively. Later, in vitro metabolites and adducts of RBC were characterized from rat liver microsomes using LC-MS/MS. Subsequently, in silico data was used as a guide for the in vitro work. Finally, in silico toxicity assessment of RBC metabolites was carried out using DEREK software and structural modification was proposed to reduce their side effects and to validate the bioactivation pathway theory using the StarDrop DEREK module. In vitro phase I metabolic profiling of RBC was performed utilizing rat liver microsomes (RLMs). Generation of reactive metabolites was investigated using potassium cyanide (KCN) as a trapping nucleophile for the transient and reactive iminium intermediates to form a stable cyano adduct that can be identified and characterized using mass spectrometry. Nine phase I metabolites and one cyano adduct of RBC were characterized. The proposed metabolic pathways involved in generation of these metabolites are hydroxylation, oxidation and reduction. The reactive intermediate generation mechanism of RBC may provide an explanation of its adverse reactions. Aryl piperazine is considered a structural alert for toxicity as proposed by the DEREK report. We propose that the generation of only one reactive metabolite of RBC in a very small concentration is due to the decreased reactivity of the piperazine ring compared to previous reports of similar drugs. Docking analysis was performed for RBC and its proposed derivatives at the active site of the human CDK6 enzyme. Methyl-RBC exhibited the best ADMET and docking analysis and fewer side effects compared to RBC and fluoro-RBC. Further drug discovery studies can be conducted taking into account this concept allowing the development of new drugs with enhanced safety profiles that were confirmed by using StarDrop software. To the best of our knowledge, this is the first literature report of RBCin vitro metabolic profiling and structural characterization and toxicological properties of the generated metabolites.

Nine phase I metabolites and one product of KCN trapping of RBC were characterized. Aryl piperazine is considered a structural alert for toxicity as proposed by the DEREK report. Methyl-RBC exhibited less toxicity and more binding affinity to CDK6.     

Odontogenic tumors: A Retrospective Study in Egyptian population using WHO 2017 classification

Background Odontogenic tumors (OTs) are considered important among oral lesions because of their clinicopathological heterogeneity and variable biological behavior. The purpose of this retrospective cross-sectional study was to evaluate the frequency and distribution of different types of odontogenic tumors based on the current 2017 WHO Classification of Head and Neck Tumors over a period of 5 years. This was achieved by reviewing the records of Cairo''s educational hospitals and institutions and comparing the results with findings in the literature.Material and Methods The records of patients diagnosed with odontogenic tumors were obtained from six educational hospitals and a single institute in Cairo which included: Oral and Maxillofacial Pathology Department, Faculty of Dentistry, Cairo University; General Pathology Department, Faculty of Medicine, Cairo University; Oral Pathology Department, Faculty of Dentistry, Ain Shams University; Eldemerdash Hospital, Ain Shams University; El-Sayed Galal Hospital, Al-Azhar University; Ahmed Maher Teaching Hospital and National Cancer Institute. These records were reviewed over a 5-year (2014-2018) period and the odontogenic tumors were investigated for frequency, age, gender and site. The data were recorded, then analyzed using SPSS software.Results Intraosseous (central) odontogenic tumors constituted 2.56% of all 8974 registered oral and maxillofacial biopsies. A total of 230 cases of OTs were collected and reviewed. Of these, 97.8% were benign and 2.17% were malignant. The mandible was the most commonly affected anatomic location. Ameloblastoma, with a predilection for the posterior mandible, was the most frequent odontogenic tumor (55.65%), followed by cemento-ossifying fibroma (14.78%) and odontoma (9.13%). Females were more commonly affected than males. Most of the patients were in the third and fourth decades of life. There were no peripheral odontogenic tumors diagnosed in this period.Conclusions Some similarities and differences between our findings and those of previous studies of various populations were witnessed. OTs may greatly diverge according to the version of the classification used and by the sample size of the study. Retrospective analysis of the relative frequency of OTs in different countries will be helpful in enhancing the understanding of OTs, which is important for both oral maxillofacial surgeons and pathologists. Key words:Odontogenic tumors, epidemiology, world health organization classification, oral pathology.     

Effects of dexamethasone and transient malnutrition on rabbits infected with aerosolized Mycobacterium tuberculosis CDC1551

Malnutrition is common in the developing world, where tuberculosis is often endemic. Rabbits infected with aerosolized Mycobacterium tuberculosis that subsequently became inadvertently and transiently malnourished had compromised cell-mediated immunity comparable to that of the rabbits immunosuppressed with dexamethasone. They had significant leukopenia and reduced delayed-type hypersensitivity responses. Malnutrition dampened cell-mediated immunity and would interfere with diagnostic tests that rely on intact CD4 T-cell responses.     

Chromosomal microarray analysis of functional Xq27-qter disomy and deletion 3p26.3 in a boy with Prader-Willi like features and hypotonia

Duplications of the long arm of the X chromosome are rare. The infantile phenotype shares some resemblance with the Prader-Willi syndrome, presenting severe psychomotor retardation, facial dysmorphic features with a broad face, a small mouth and a thin pointed nose, hypotonia, urogenital malformation and proneness to infections. We report a boy with an additional Xq27-qter chromosome segment translocated onto the short arm of chromosome 3. The karyotype was 46,XY,der(3)t(X;3)(q27.3; p26.3)mat. This cryptic unbalanced X-autosome translocation resulted in Xq27-qter functional disomy and a deletion 3p26.3. A detailed analysis of the constitutional chromosomal changes in the patient was performed using array-CGH, FISH and PCR. The aim was to characterize the size and the location of the duplication Xq27-qter (8.18 Mb) and of the deletion 3p26.3 (1.05 Mb), to establish phenotype-genotype correlations and to offer genetic counselling.