Background and purpose: New compounds and innovative therapeutic approaches are trying to prevent antimicrobial resistance, which has become a global health challenge. Experimental approach: This study includes a series of twelve mono-, di- and trichlorinated 1-hydroxynaphthalene-2-carboxanilides designed as multitarget agents. All compounds were evaluated for their antistaphylococcal activity. Furthermore, MTT assay and chemoproteomic analysis of selected compounds were performed. Cytotoxicity in human cells was also tested. Key results: N-(3,5-Dichlorophenyl)-1-hydroxynaphthalene-2-carboxamide (10) demon-strated activity comparable to or higher than clinically used drugs, with minimum inhibitory concentrations (MICs) of 0.37 μM. The compound was equally effective against clinical isolates of methicillin-resistant S. aureus. On the other hand, compound 10 showed 96 % inhibition of S. aureus respiration only at a concentration of 16× MIC. Chemoproteomic analysis revealed that the effect of agent 10 on staphylococci resulted in the downregulation of four proteins. This compound expressed no in vitro cytotoxicity up to a concentration of 30 μM. Conclusion: From the set of tested mono-, di- and trisubstituted derivatives, it is evident that the position of chlorine atoms is decisive for significant antistaphylococcal activity. Inhibition of energy metabolism does not appear to be one of the main mechanisms of action of compound 10; on the contrary, the antibacterial effect may likely be contributed by downregulation of proteins (especially ATP-dependent protease ATPase subunit HslU) involved in processes essential for bacterial survival and growth, such as protein, nucleotide/nucleic acid synthesis and efficient protein repair/degradation.

Background and purpose: Many new compounds are being prepared to overcome the problem of increasing microbial resistance and the increasing number of infections. Experimental approach: This study includes a series of twenty-seven mono-, di- and trisubstituted 2-hydroxynaphthalene-1-carboxanilides designed as multitarget agents. The compounds are substituted with methoxy, methyl, and nitro groups, as well as additionally with chlorine, bromine, and trifluoromethyl at various positions. All the compounds were evaluated for antibacterial activities against Gram-positive and Gram-negative bacteria and mycobacteria. Cytotoxicity on human cells was also tested. Key results: Three compounds showed activity comparable to clinically used drugs. N-(3,5-Dimethylphenyl)-2-hydroxynaphthalene-1-carboxamide (13) showed only anti¬sta¬phylococcal activity (minimum inhibitory concentration (MIC) = 54.9 µM); 2-hydroxy-N-[2-methyl-5-(tri¬fluoro¬methyl)phenyl]naphthalene-1-carboxamide (22) and 2-hydroxy-N-[4-nitro-3-(trifluoromethyl)phe¬nyl]na¬phtha¬lene-1-carboxamide (27) were active across the entire spectrum of tested bacteria/mycobacteria, both against the sensitive set and against resistant isolates (MICs range 0.3 to 92.6 µM). Compound 22 was even active against E. coli (MIC = 23.2 µM). The active agents showed no in vitro cytotoxicity up to a concentration of 30 μM. Conclusion: Compounds with trifluoromethyl in the meta-anilide position, experimental lipophilicity expressed as log k (logarithm of the capacity factor) in the range of 0.31 to 0.34 and calculated electron σ parameter for the anilide substituent higher than 0.59 were effective. The investigated compounds meet the definition of Michael acceptors. Based on ADME screening, the investigated compounds 13, 22 and 27 should have suitable physico¬chemical parameters for good bioavailability in the organism. Therefore, these are promising agents for further study.

Overall survival of patients classified according to the European LeukemiaNet 2020 classification. Chronic phase (CP), accelerated phase (AP), blast crisis (BC), low risk (LR), intermediate risk (IR), high risk (HR).

Blood basophils ≥ 20 percent is reportedly associated with a poor prognosis and used to define accelerated phase of chronic myeloid leukaemia (CML) in some classifications. However, quantification of blood basophils is by percentage is inaccurate. Using a Patient Similarity Network (PSM) approach we identified basophil concentration rather than percentage as the more accurate predictive co-variate. To test this observation we interrogated data for a possible correlation between blood basophils quantified by concentration in a training cohort of 131 subjects with newly-diagnosed chronic phase CML receiving tyrosine kinase-inhibitor (TKI)-therapy. Subjects with a basophil concentration ≥ 12.2 x 10E + 9/L had poorer event-free survival (EFS, Odds Ratio [OR] = 12.3 [95% Confidence Interval [CI]. 4.2, 36.1]; p < 0.0001) and failure-free survival (FFS; OR = 10.4 [3.89, 27.72]; p < 0.0001). TKI switch-free survival and progression-free survival were also correlated with basophil concentration. The negative impact of a high basophil concentration was validated in an independent cohort of 1,870 subjects. We explain why basophil concentration is a more accurate co-variate. Our data indicate blood basophil concentration at diagnosis rather than percentage is a more accurate predictor of outcomes in persons with newly-diagnosed chronic phase CML receiving TKI-therapy.

Background A lower dosage of tyrosine kinase inhibitors (TKIs) in patients with chronic myeloid leukaemia (CML) has shown efficacy in managing short-term toxicity and maintaining a deep molecular response in patients who fail to achieve treatment-free remission. Method From over 700 patients with CML who were treated at two centres over the last three decades, this retrospective study identified eight patients characterised by long-term treatment failure and simultaneous prolonged significant haematologic toxicity that prevented the use of the standard tyrosine kinase inhibitor dosage. Results Patients had a high or intermediate ELTS risk score, and most had significant comorbidities. Two patients were treated previously with busulfan, and four were aged over 70, which might explain the reduced pool of normal haematopoietic stem cells. However, concomitant myelodysplastic syndrome or the presence of clonal haematopoiesis of indeterminate potential was not demonstrated. Despite prolonged treatment failure, the survival of these patients (who were ineligible for stem cell transplantation) ranged from 45-396 months. Neither mutations in the ABL kinase domain nor additional cytogenetic abnormalities developed during the treatment of these patients, prompting speculation about the low selective pressure of low-dose tyrosine kinase inhibitors and/or the absence of mutations at diagnosis. Conclusion It is important not to stop treatment with tyrosine kinase inhibitors at a low personalised dosage in CML patients with prolonged significant haematologic toxicity despite long-term treatment failure.

Background Basophilia is common in chronic myeloid leukaemia (CML) and is associated with a poor prognosis. Previously it was quantified by percentage and a binary < or ≥ 20%. However, blood basophil concentration at diagnosis may be a better prognostic co-variate. Methods The hypothesis generating cohort was a 135 newly-diagnosed subjects with BCR::ABL1-positive CML receiving a tyrosine kinase-inhibitor (TKI). Median follow-up was 6.3 years (range, 4-16 years). Data were analyzed by Kaplan-Meier curves and multi-variable patient similarity network (PSN). A cohort of 1919 subjects was used to validate the test hypothesis. Subject gave informed consent for non-interventional data collection and the study was approved by Ethics Committees. Results Multi-variable PSNs of the hypothesis generating cohort indicated 5 subject clusters. Incidence of TKI switch was highest in clusters 1 (45%), 5 (40%) and 4 (30%) which were characterized by the increased basophil concentrations at diagnosis compared with clusters 2 (13%) and 3 (14%) with the low basophil concentrations at diagnosis. PSNs identified the combination of basophil concentration ≥ 8x10E+9/L, basophil percentage ≥ 5 % and WBC ≥ 164x10E+9/L to be associated with significantly worse FFS (p < 0.001) and PFS ( p = 0.02) but not survival ( p = 0.18). Basophil concentration ≥ 8x10E+9/L and WBC concentration ≥ 164x10E+9/L were associated with worse EFS (p < 0.001), (FFS (p < 0.001; Figure 1) but not PFS or survival. Basophil percentage at diagnosis had no impact on outcomes using ≥ 5% or ≥ 20% cutoffs. In the validation cohort the combination of basophil concentration ≥ 8x10E+9/L, basophil percentage ≥ 5% and WBC ≥ 164x10E+9/L at diagnosis was associated with worse FFS ( p < 0.001) but not PFS or survival. Basophils ≥ 5% was associated with worse survival ( p = 0.04) and ≥ 20% with worse FFS ( p = 0.04) and survival ( p = 0.009). Basophil concentration ≥ 8×10E+9/L was associated with worse PFS ( p < 0.001; Figure 1) and survival ( p = 0.03). There was no effect of WBC concentration on any outcome. Conclusion We show blood basophil concentration at diagnosis correlates with diverse outcomes in newly-diagnosed persons with CML receiving TKI-therapy. Support IGA_LF_2023_05, MH_CZ-DRO (FNOL, 00098892)

Background For decades, CML has been considered to be a triphasic disease recognizing 3 distinct stages: chronic phase (CP), accelerated phase (AP), and blast crisis (BC). Since the discovery of TKIs, the prognosis of CML patients has rapidly improved and less than 10% of patients diagnosed in CP experience disease progression on the therapy. Based on low incidence of CML progression and the fact that CML biological behavior seems biphasic, the new WHO 2022 classification of CML deemed AP less relevant and suggested the omission of AP. Many experts, however, still advocate for its inclusion in CML classification (e.g. ELN 2020, ICC 2022, NCCN 2023), even though the universal definition of AP is not established and differs in between publications. Methods This retrospective, real-world study is based on the Czech nationwide CML registry INFINITY with the data of newly diagnosed CML patients who agreed and provided their written consent. Recruited subjects were adult patients diagnosed in years 2005 - 2022 with sufficient data to determine the phase of the disease at diagnosis and follow-ups to assess their overall survival (OS), disease specific survival (DSS) and progression-free survival (PFS). The phase of the disease at the time of diagnosis was determined using the classification according to ELN 2020, ICC 2022, WHO 2016, and WHO 2022. Whenever CP was established, EUTOS Long-Term Survival (ELTS) score was calculated, assigning the patient to low risk (LR), intermediate risk (IR) and high risk (HR) group. OS and PFS are defined according to published guidelines. DSS is defined as the time from diagnosis to death due to CML disease or CML treatment. Results During the studied time period, 1660 new cases of CML were reported in INIFINITY registry. Of them, 1500 patients had data sufficient to classify the phase of the disease at the time of the diagnosis according to ELN 2020 guidelines and WHO 2022 classification and 1395 patients had data to assess the phase according to WHO 2016 and ICC 2022 classifications. When using ELN 2020 guidelines, groups were assigned as follows: LR CP- 784 patients (52.3%), IR CP- 421 patients (28.1%), HR CP- 227 patients (15.1%), AP- 42 patients (2.8%), BC- 26 patients (1.7%). There were significant differences in representation by gender, age, comorbidities and performance score amongst patient groups. When comparing just HR CP and AP CML patients, there were no significant differences. Calculated estimates of 5- and 10- year OS, respectively, were 92.5% and 87.2% for LR CP, 83.5% and 65.9% for IR CP, 76.9% and 65.5% for HR CP, 59.2% and 45.9% for AP, and 32.9% for BC, p= 0.031 for HR CP versus AP (Figure 1). In the case of WHO 2016 and ICC 2022 classifications, 714 (51.2%) patients were diagnosed in LR CP, 374 (26.8%) in IR CP, 154 (11.0%) in HR CP, 125 (9.0%) in AP, and 28 (2.0%) in BC. Again, there were significant differences in representation by gender, age, comorbidities and performance score amongst patient groups. When comparing just HR CP and AP CML patients, only age at the time of diagnosis was significantly different. Calculated estimates of 5- and 10- year OS, respectively, were 93.2% and 87.5% for LR CP, 84.9% and 67.7% for IR CP, 80.0% and 69.5% for HR CP, 65.7% and 55.6% for AP, and 39.7% for BC. We also performed propensity score matching according to age for patients in HR CP and AP and calculated estimates of 5- and 10- year OS, respectively, were 85.4% and 74.4% for HR CP and 60.2%, and 52.0% for AP, p= 0.003 (Figure 2). Similar results were obtained when testing for PFS and DSS. Summary Real-world data obtained from 1500 patients diagnosed in Czechia over 17 years, in our opinion, support the need to recognize the existence of AP at the time of diagnosis, even though a singular definition of AP is not agreed upon. Patients in AP have worse survival scores (OS, DSS, PFS) compared to patients in CP and/or HR CP. Moreover, because ELTS risk score was calculated on patients in CP, it may not be suitable for patients in AP, which would be changed to CP according to the new WHO 2022 classification. In conclusion, based on the real-world data with long-term follow-up, we believe it would be reasonable to keep the AP as part of CML classification of newly diagnosed patients. This publication was supported by the grant number MUNI/A/1224/2022, Programme EXCELES, ID Project No. LX22NPO5102, and by the Ministry of Health of the Czech Republic grant number 00023736.

The treatment outcome in patients with chronic myeloid leukaemia (CML) in blast crisis (BC) is unsatisfactory despite the use of allogeneic stem cell transplantation (ASCT). Moreover, in some patients ASCT is contraindicated, with limited treatment options. We report the case series of two patients with lymphoid BC CML in whom ASCT was not approachable. The first patient developed BC two months after diagnosis in association with dic(7;9)(p11.2;p11.2) and T315I mutation. Blast crisis with central nervous system leukemic involvement and K611N mutation of the SETD2 gene developed abruptly in the second patient five years after ceasing treatment with nilotinib in major molecular response (MMR) at the patient’s request. Both underwent one course of chemotherapy in combination with rituximab and imatinib, followed by dasatinib and interferon α (INFα) treatment in the first and dasatinib alone in the second case. Deep molecular response (DMR; MR 4.0) was achieved within a short time in both cases. It is probable that DMR was caused by a specific immune response to CML cells, described in both agents. The challenging medical condition that prompted these case series, and the subsequent results, suggest a re-visit to the use of a combination of well-known drugs as an area for further investigation.

Topic: 4. Acute myeloid leukemia - Clinical Background: At diagnosis, acute promyelocytic leukaemia (APL) is associated with high risk of bleeding complications due to coagulopathy and thrombocytopenia. In current guidelines, it is explicitly stated that the introduction of the central venous catheter should be avoided in this initial phase. However, patients absolutely need high-quality venous access for the administration of both antitumor and supportive treatment. A peripherally inserted central catheter (PICC) is a central venous catheter intended for medium to long-term use. Its implantation and use are safe in the hands of experienced personnel. We started the PICC implementation program at our institution in 2016. After gaining the necessary experience, we also started implanting this type of catheter in patients with acute leukaemia. Due to the lower risk of bleeding complications associated with implantation, we decided to prefer PICC in patients with APL as well. Aims: Retrospective analysis of a single institution experience with PICC implantation and use in all consecutive patients with newly diagnosed APL over the past 5 years. Methods: We have treated a total of 13 patients with newly diagnosed APL since 2017. A PICC was used in all of them from the beginning of the induction treatment. There were 8 women and 5 men with an median age of 60 (24-73) years. The first five patients were treated with a combination of ATRA and chemotherapy, the others according to the ATRA-ATO protocol. The selection of the treatment protocol was determined by the currently valid recommendation and reimbursement rules of the treatment within the framework of the health insurance. The PICC was usually implanted on the second day of hospitalization (7 patients), less often on the first day (3 patients). In the first two patients in the group, the implantation was performed on the 6th and 5th day, respectively. The punctured vein was the brachial or basilic vein in 10 cases, and the axillary vein (with longer subcutaneous tunnelling) in 3 patients. A 2-lumen 5F catheter was used more often (7 times – including in all 5 patients treated with chemotherapy), a 1-lumen 4F catheter was inserted 6 times. The median level of platelets at implantation was 55 (20 – 118) x 109/l. All patients had coagulopathy typical of APL. Results: In 7 patients, the catheter was used for the entire duration of the treatment and was extracted after its completion. The median duration of use was 202 (159-298) days. Three patients continue treatment with the catheter used 116, 151 and 262 days. In two patients, the catheter was extracted before the end of treatment. In the first patient of the group after 30 days for local inflammation at the entry site, in the seventh patient after 31 days due to fever of unknown origin. One patient died during induction treatment (ATRA/ATO) from respiratory failure caused by COVID-19 pneumonia. In addition to the above, we noticed only one more complication, which did not require catheter extraction. It was a short partial thrombosis of the vein around the catheter 26 days after implantation. None of the patients experienced complication during catheter implantation. Summary/Conclusion: Our 5-year experience suggests that in the hands of an experienced team, the PICC is a safe device providing long-term central venous access for the care of patients with acute promyelocytic leukaemia, including the most critical initial phase of treatment. Supported by IGA_LF_UP_2023_005 a MZ ČR – RVO (FNOL, 00098892). Keywords: Acute promyelocytic leukemia, Central venous catheter

A series of thirty-two anilides of 3-(trifluoromethyl)cinnamic acid (series 1) and 4-(trifluoromethyl)cinnamic acid (series 2) was prepared by microwave-assisted synthesis. All the compounds were tested against reference strains Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212 and resistant clinical isolates of methicillin-resistant S. aureus (MRSA) and vancomycin-resistant E. faecalis (VRE). All the compounds were evaluated in vitro against Mycobacterium smegmatis ATCC 700084 and M. marinum CAMP 5644. (2E)-3-[3-(Trifluoromethyl)phenyl]-N-[4-(trifluoromethyl)phenyl]prop-2-enamide (1j), (2E)-N-(3,5-dichlorophenyl)-3-[3-(trifluoromethyl)phenyl]prop-2-enamide (1o) and (2E)-N-[3-(trifluoromethyl)phenyl]-3-[4-(trifluoromethyl)-phenyl]prop-2-enamide (2i), (2E)-N-[3,5-bis(trifluoromethyl)phenyl]-3-[4-(trifluoromethyl)phenyl]-prop-2-enamide (2p) showed antistaphylococcal (MICs/MBCs 0.15–5.57 µM) as well as anti-enterococcal (MICs/MBCs 2.34–44.5 µM) activity. The growth of M. marinum was strongly inhibited by compounds 1j and 2p in a MIC range from 0.29 to 2.34 µM, while all the agents of series 1 showed activity against M. smegnatis (MICs ranged from 9.36 to 51.7 µM). The performed docking study demonstrated the ability of the compounds to bind to the active site of the mycobacterial enzyme InhA. The compounds had a significant effect on the inhibition of bacterial respiration, as demonstrated by the MTT assay. The compounds showed not only bacteriostatic activity but also bactericidal activity. Preliminary in vitro cytotoxicity screening was assessed using the human monocytic leukemia cell line THP-1 and, except for compound 2p, all effective agents did show insignificant cytotoxic effect. Compound 2p is an interesting anti-invasive agent with dual (cytotoxic and antibacterial) activity, while compounds 1j and 1o are the most interesting purely antibacterial compounds within the prepared molecules.

Background: Despite the elevated basophils at diagnosis may have a prognostic value in chronic myeloid leukemia (CML), there is limited information about the clinical relevance of the changes in relative and absolute peripheral blood basophils counts during the initial cytoreduction treatment in newly diagnosed CML. Methods: This study included a real-world cohort of newly diagnosed 136 BCR-ABL1+ CML patients (56 females/80 males, median age of 58 years, range: 24-92 years) indicated for the first-line tyrosine kinase inhibitor (TKI) treatment. Blood counts and laboratory parameters were assessed at the diagnosis and during first month of cytoreduction treatment with hydroxyurea and/or first line TKI (imatinib or nilotinib in 11 cases). The median follow-up of the patients was 6.3 years (range: 3.5 months - 15.7 years). Kaplan-Mayer curves, odds ratio including 95% confidence intervals (CI) and receiver operation curves (ROC) were calculated using R software. Results: At diagnosis, there was high interindividual heterogeneity in basophil percentage (median 4%, range 0-45%) and basophil absolute counts (median 4.6, range 0-67.6x 109/L). During the first months of cytoreduction treatment, 65 (47.8%) patients had a decrease or stable values in basophil percentage and 71 (52.2%) had increased basophil percentages comparing to the diagnosis. Elevated absolute basophil counts after cytoreduction was evident in 26 (19%) patients, predominantly in young males, with high risk Sokal score, splenomegaly, high leukocyte counts and increased lactate dehydrogenase levels at the diagnosis. The higher percentage (>15%) of basophils at diagnosis was associated with a later cytogenetic response (p=0.011), however we did not prove the prognostic impact for achieving treatment response of relative basophils counts after cytoreduction on overall, even at levels >20%. However, patients with lower absolute counts of basophils at diagnosis had a lower risk for introduction of the second or higher lines of TKI treatment (odds ratio 0.109, 95%CI 0.044-0.270), using absolute basophil count of 12.2x109/L as a threshold revealed from ROC (AUC 0.733). Similar results were observed for absolute counts (0.463, 0.217-0.984) and relative counts (>15%, 0.371, 0.143-0.963) of basophils after cytoreduction. Our pilot data suggest that the increase in relative basophilia in CML patients during cytoreduction may be linked to a slower clearance of basophils from peripheral blood comparing to the neutrophils. Conclusion: Our study on real-world cohort CML patients showed that progression of relative basophilia is a common phenomenon during the cytoreduction and it has no independent prognostic value for achievement of treatment response and survival of patients. However, our data suggested that a high absolute basophil counts at the diagnosis of CML and after the cytoreduction treatment are associated with the risk for a subsequent change in the TKI treatment. Future studies focused on absolute basophil counts in larger cohorts of CML patients are warranted to confirm our results. Supported by MH CZ - DRO (FNOl, 00098892), IGA_LF_2022_001.

A series of twenty-two monosubstituted N-aryl-4-hydroxyquinoline-3-carboxanilides designed as dual anti-invasive agents was prepared and characterized. Lipophilicity significantly affects biological activities of compounds and ADME properties; therefore, the lipo-hydrophilic properties of these 4-hydroxyquinoline-3-carboxanilides were investigated. All the derivatives were analyzed using reversed-phase high-performance liquid chromatography. The procedure was carried out under isocratic conditions with methanol as the organic modifier in the mobile phase using an end-capped non-polar C18 stationary reversed-phase column. In this study, correlations between the logarithm of the capacity factor k and log P/Clog P values calculated using various methods are discussed, as well as the relationships between lipophilicity and chemical structure of the studied compounds.

A series of eighteen 4-chlorocinnamanilides and eighteen 3,4-dichlorocinnamanilides were designed, prepared and characterized. All compounds were evaluated for their activity against gram-positive bacteria and against two mycobacterial strains. Viability on both cancer and primary mammalian cell lines was also assessed. The lipophilicity of the compounds was experimentally determined and correlated together with other physicochemical properties of the prepared derivatives with biological activity. 3,4-Dichlorocinnamanilides showed a broader spectrum of action and higher antibacterial efficacy than 4-chlorocinnamanilides; however, all compounds were more effective or comparable to clinically used drugs (ampicillin, isoniazid, rifampicin). Of the thirty-six compounds, six derivatives showed submicromolar activity against Staphylococcus aureus and clinical isolates of methicillin-resistant S. aureus (MRSA). (2E)-N-[3,5-bis(trifluoromethyl)phenyl]- 3-(4-chlorophenyl)prop-2-enamide was the most potent in series 1. (2E)-N-[3,5-bis(Trifluoromethyl)phenyl]-3-(3,4-dichlorophenyl)prop-2-enamide, (2E)-3-(3,4-dichlorophenyl)-N-[3-(trifluoromethyl)phenyl]prop-2-enamide, (2E)-3-(3,4-dichloro- phenyl)-N-[4-(trifluoromethyl)phenyl]prop-2-enamide and (2E)-3-(3,4-dichlorophenyl)- N-[4-(trifluoromethoxy)phenyl]prop-2-enamide were the most active in series 2 and in addition to activity against S. aureus and MRSA were highly active against Enterococcus faecalis and vancomycin-resistant E. faecalis isolates and against fast-growing Mycobacterium smegmatis and against slow-growing M. marinum, M. tuberculosis non-hazardous test models. In addition, the last three compounds of the above-mentioned showed insignificant cytotoxicity to primary porcine monocyte-derived macrophages.

Pattern 1-hydroxy-N-(2,4,5-trichlorophenyl)-2-naphthamide and the thirteen original carbamates derived from it were prepared and characterized. All the compounds were tested against Staphylococcus aureus ATCC 29213 as a reference and quality control strain and in addition against three clinical isolates of methicillin-resistant S. aureus (MRSA). Moreover, the compounds were evaluated against Enterococcus faecalis ATCC 29212, and preliminary in vitro cytotoxicity of the compounds was assessed using the human monocytic leukemia cell line (THP-1). The lipophilicity of the prepared compounds was experimentally determined and correlated with biological activity. While pattern anilide had no antibacterial activity, the prepared carbamates demonstrated high antistaphylococcal activity comparable to the used standards (ampicillin and ciprofloxacin), which unfortunately were ineffective against E. feacalis. 2-[(2,4,5-Trichlorophenyl)carba- moyl]naphthalen-1-yl ethylcarbamate (2) and 2-[(2,4,5-trichlorophenyl)carbamoyl]naphthalen-1-yl butylcarbamate (4) expressed the nanomolar minimum inhibitory concentrations (MICs 0.018–0.064 μM) against S. aureus and at least two other MRSA isolates. Microbicidal effects based on the minimum bactericidal concentrations (MBCs) against all the tested staphylococci were found for nine carbamates, while 2-[(2,4,5-trichlorophenyl)carbamoyl]naphthalen-1-yl heptylcarbamate (7) and 2-[(2,4,5-trichlorophenyl)carbamoyl]naphthalen-1-yl (4-phenylbutyl)carbamate (14) demonstrated MBCs in the range of 0.124–0.461 μM. The selectivity index (SI) for most investigated carbamates was >20 and for some derivatives even >100. The performed tests did not show an effect on the damage to the bacterial membrane, while the compounds were able to inhibit the respiratory chain of S. aureus.

Unsubstituted (2E)-N-phenyl-3-[2-(trifluoromethyl)phenyl]prop-2-enamide and six other ortho- or para-halogen-substituted anilides of 2-(trifluoromethyl)cinnamic acid were prepared. As the benzene nucleus of cinnamic acid itself is substituted in C(2) position with a trifluoromethyl moiety that is spatially close to both the amide bond and the halogen (F, Cl, CF3) ortho-substitution of the anilide ring, interesting intramolecular interactions can be expected. Other derivatives are substituted at the para-position of the anilide ring, so that intermolecular interactions can be expected. Thus, it can be assumed that predicted properties, especially lipophilicity, will differ significantly from experimentally determined values. All the discussed compounds were analyzed using the reversed-phase high-performance liquid chromatography method. The procedure was performed under isocratic conditions with methanol as an organic modifier in the mobile phase using an end-capped non-polar C18 stationary reversed-phase column. In the present study, the structure–lipophilicity relationships of the studied compounds are discussed.

Lipophilicity is one of the important properties of bioactive molecules, according to which the nature of a potential drug is assessed. Evaluation of the lipophilicity of selected cinnamic acid derivatives was performed by high-performance liquid chromatography (HPLC) using reversed (RP) stationary phase C18 and under isocratic conditions. In the case of determining the capacity factor k, methanol and water were applied to the system as the mobile phase. The distribution coefficient D was determined using a mobile phase composed of methanol and acetate buffer (pH 7.4 or pH 6.5) to ensure a constant pH. This contribution aims to compare the influence of various factors on the lipophilicity of selected trifluoromethyl substituted cinnamanilides, including pH and the position and nature of specific substituents in the anilide portions of the molecules. The results of this study will then be used to evaluate structure-lipophilicity relationships, druglikeness, and structure-activity relationships. Acknowledgement: This study was supported by a grant project of the Comenius University in Bratislava, Slovakia (UK/228/2021) and by the Slovak Research and Development Agency (APVV-17-0373).

Inflammatory and oncogenic signaling, both known to challenge genome stability, are key drivers of BCR-ABL-positive chronic myeloid leukemia (CML) and JAK2 V617F-positive chronic myeloproliferative neoplasms (MPNs). Despite similarities in chronic inflammation and oncogene signaling, major differences in disease course exist. Although BCR-ABL has robust transformation potential, JAK2 V617F-positive polycythemia vera (PV) is characterized by a long and stable latent phase. These differences reflect increased genomic instability of BCR-ABL-positive CML, compared to genome-stable PV with rare cytogenetic abnormalities. Recent studies have implicated BCR-ABL in the development of a "mutator" phenotype fueled by high oxidative damage, deficiencies of DNA repair, and defective ATR-Chk1-dependent genome surveillance, providing a fertile ground for variants compromising the ATM-Chk2-p53 axis protecting chronic phase CML from blast crisis. Conversely, PV cells possess multiple JAK2 V617F-dependent protective mechanisms, which ameliorate replication stress, inflammation-mediated oxidative stress and stress-activated protein kinase signaling, all through up-regulation of RECQL5 helicase, reactive oxygen species buffering system, and DUSP1 actions. These attenuators of genome instability then protect myeloproliferative progenitors from DNA damage and create a barrier preventing cellular stress-associated myelofibrosis. Therefore, a better understanding of BCR-ABL and JAK2 V617F roles in the DNA damage response and disease pathophysiology can help to identify potential dependencies exploitable for therapeutic interventions.