Corticosteroid-associated osteonecrosis of the femoral head in an 87-year-old patient following treatment for COVID-19-associated pneumonia: A case report.

Coronavirus disease 2019 is known to cause severe acute respiratory syndrome, and serious cases need to be treated with corticosteroids. Herein, we report an 87-year-old woman who developed bilateral osteonecrosis of the femoral head after corticosteroid treatment for coronavirus disease 2019-related pneumonia. Sixteen months after treatment, she developed right hip pain without any evidence of trauma. A diagnosis of bilateral osteonecrosis of the femoral head was made based on sclerotic bands on plain radiographs and low-signal bands on T1-weighted magnetic resonance images. The patient underwent right total hip arthroplasty 4 months after symptom onset. Histological examination of the resected femoral head revealed pathological evidence of osteonecrosis. The postoperative course was good, and the patient can now walk unassisted. To the best of our knowledge, this is the first report of histologically proven osteonecrosis after corticosteroid therapy for coronavirus disease 2019-related disease.

The role of torsional stress in the development of subchondral insufficiency fracture of the femoral head: A finite element model analysis.

BACKGROUND: Subchondral insufficiency fracture of the femoral head generally occurs without evidence of trauma or with a history of minor trauma. Insufficient bone quality is considered one cause; however, the detailed mechanism of fracture development at the subchondral area (SA) is not understood. The aim of this study was to clarify the directions of force that cause subchondral fracture using finite element model analysis. METHODS: Two types of finite element models were generated from the CT data of femurs obtained from three individuals without osteoporosis (normal models) and another three with osteoporosis (osteoporosis models). Three directions of force, including compressive, shearing, and torsional, were applied to the femoral head. The distribution of von Mises stress (Mises stress) was evaluated at the SA, principal compressive trabeculae (PC), and principal tensile trabeculae. RESULTS: Under compressive force, the mean Mises stress value was greatest at the PC in both the normal and osteoporosis models. Under shearing force, the mean Mises stress value tended to be greatest at the SA in the normal model and at the PC in the osteoporosis model. Under torsional force, the mean Mises stress value was greatest at the SA in both types of models. CONCLUSIONS: The torsional force showed the greatest Mises stress at the SA in both the normal and osteoporosis models, suggesting the importance of torsion as a possible force responsible for subchondral insufficiency fracture development.

Side-to-side variability in the femoral neck anteversion angle: A study of the Japanese population with osteonecrosis of the femoral head.

BACKGROUND: Side-to-side variability in the femoral neck anteversion angle (FA) reportedly varies from 0.0° to 17.3°. To investigate the side-to-side variability in the FA and the relationship between the FA and the morphology of the acetabulum in the Japanese population, we performed a three-dimensional computed tomography (CT)-based study involving patients with osteonecrosis of the femoral head (ONFH). METHODS: CT data were obtained from 170 nondysplastic hips of 85 patients with ONFH. The FA and acetabular coverage parameters, including the acetabular anteversion angle, acetabular inclination angle, and acetabular sector angle in the anterior, superior, and posterior directions, were measured using three-dimensional CT. The distribution of the side-to-side variability in the FA was evaluated separately for each of the five degrees. RESULTS: The mean side-to-side variability in the FA was 6.7° ± 5.3° (range, 0.2°-26.2°). The distribution of the side-to-side variability in the FA was 0.0°-5.0° in 41 patients (48.2%), 5.1°-10.0° in 25 patients (29.4%), 10.1°-15.0° in 13 patients (15.3%), 15.1°-20.0° in 4 patients (4.7%), and >20.1° in 2 patients (2.4%). There was a weak negative correlation between the FA and anterior acetabular sector angle (r = -0.282, P < 0.001) and a very weak positive correlation between the FA and acetabular anteversion angle (r = 0.181, P < 0.018). CONCLUSIONS: The mean side-to-side variability in the FA was 6.7° ± 5.3° (range, 0.2°-26.2°) in Japanese nondysplastic hips, and about 20% of the patients had a side-to-side variability of >10°.

Assessment of the Hip Flexor Muscles Shortly after Curved Periacetabular Osteotomy using Magnetic Resonance Imaging.

BACKGROUND: Curved periacetabular osteotomy requires detachment and retraction of the hip flexors. In this study, we evaluated hip flexor muscle status by magnetic resonance imaging (MRI) shortly after curved periacetabular osteotomy. METHODS: We retrospectively evaluated 60 hips of 56 patients by MRI 1 week and 3 months after curved periacetabular osteotomy performed from August 2017 to December 2019. We classified the condition of the flexors as follows: Grade 0, normal; Grade I, strain/edema; Grade II, partial tear; and grade III, complete tear. RESULTS: At 1 week after surgery, the iliacus muscle was classified as grades I and II in 12.0 and 88.0% of hips; psoas as grades 0, I and II in 22.0, 72.0, and 6.0%; sartorius muscle as grades 0, I and II in 6.0, 62.0, and 32.0%; and rectus femoris muscle as grades 0 and I in 86.0 and 14.0%, respectively. At 3 months, 82.0, 88.0, and 96.0% of psoas, sartorius, and rectus femoris muscles, respectively, had improved to grade 0, whereas the iliacus was grades I and II in 94.0 and 6.0%, respectively. These changes in the iliacus muscle at 3 months were not significantly associated with patient characteristics, radiographic data, or clinical scores. CONCLUSIONS: All the iliacus, 78% of psoas, 94% of sartorius, and 14% of rectus femoris muscles appeared abnormal on MRI 1 week after curved periacetabular osteotomy. However, at 3 months, only 18% of psoas, 12% of sartorius, and 4% of rectus femoris muscles appeared abnormal, whereas all iliacus muscles still appeared abnormal. These abnormalities did not significantly affect clinical scores.

Mechanical Influence of Pubic Nonunion on the Stress Distribution After Curved Periacetabular Osteotomy: Patient-Specific Three-Dimensional Finite Element Analysis.

BACKGROUND: Pubic nonunion after curved periacetabular osteotomy (CPO) reportedly occurs in 1%-17% of patients and causes pubic pain in 21%. Furthermore, pubic nonunion is associated with a risk of ischial ramus stress fracture, but the mechanical influence of pubic nonunion has not been fully clarified. METHODS: Patient-specific finite element (FE) analysis was performed using Mechanical Finder software. Three FE models (pre-CPO, union, and nonunion models) were constructed from preoperative and postoperative computed tomographic data. The contact area (mm2) and contact pressure (MPa) in the hip joint as well as the equivalent stress (MPa) at the ischial ramus were evaluated among the 3 FE models. RESULTS: Patient-specific FE models were generated using 18 consecutive hips treated with CPO. The mean contact pressure in the hip joint was not significantly different between the union and nonunion models (0.50 ± 0.10 vs 0.50 ± 0.09 MPa, P = .88). However, the mean equivalent stress at the ischial ramus in the nonunion models was 1.7 times higher than that in the union models (1.13 ± 0.77 vs 0.64 ± 0.45 MPa, P < .01). CONCLUSION: FE analysis revealed that pubic nonunion did not affect the mechanical distribution in the hip joint itself but increased the mean equivalent stress at the ischial ramus. This finding suggests the importance of achieving pubic union after CPO to avoid the risk of ischial ramus stress fracture.

Acid ceramidase promotes drug resistance in acute myeloid leukemia through NF-κB-dependent P-glycoprotein upregulation.

Acute myeloid leukemia (AML) is the most common acute leukemia in adults. More than half of older AML patients fail to respond to cytotoxic chemotherapy, and most responders relapse with drug-resistant disease. Failure to achieve complete remission can be partly attributed to the drug resistance advantage of AML blasts that frequently express P-glycoprotein (P-gp), an ATP-binding cassette transporter. Our previous work showed that elevated acid ceramidase (AC) levels in AML contribute to blast survival. Here, we investigated P-gp expression levels in AML relative to AC. Using parental HL-60 cells and drug-resistant derivatives as our model, we found that P-gp expression and efflux activity were highly upregulated in resistant derivatives. AC overexpression in HL-60 conferred resistance to the AML chemotherapeutic drugs, cytarabine, mitoxantrone, and daunorubicin, and was linked to P-gp upregulation. Furthermore, targeting AC through pharmacologic or genetic approaches decreased P-gp levels and increased sensitivity to chemotherapeutic drugs. Mechanistically, AC overexpression increased NF-κB activation whereas NF-kB inhibitors reduced P-gp levels, indicating that the NF-kappaB pathway contributes to AC-mediated modulation of P-gp expression. Hence, our data support an important role for AC in drug resistance as well as survival and suggest that sphingolipid targeting approaches may also impact drug resistance in AML.

Role of deltaNp63(pos)CD44v(pos) cells in the development of N-nitroso-tris-chloroethylurea-induced peripheral-type mouse lung squamous cell carcinomas.

The role of cells expressing stem cell markers deltaNp63 and CD44v has not yet been elucidated in peripheral-type lung squamous cell carcinoma (pLSCC) carcinogenesis. Female A/J mice were painted topically with N-nitroso-tris-chloroethylurea (NTCU) for induction of pLSCC, and the histopathological and molecular characteristics of NTCU-induced lung lesions were examined. Histopathologically, we found atypical bronchiolar hyperplasia, squamous metaplasia, squamous dysplasia, and pLSCCs in the treated mice. Furthermore, we identified deltaNp63(pos)CD44v(pos)CK5/6(pos)CC10(pos) clara cells as key constituents of early precancerous atypical bronchiolar hyperplasia. In addition, deltaNp63(pos)CD44v(pos) cells existed throughout the atypical bronchiolar hyperplasias, squamous metaplasias, squamous dysplasias, and pLSCCs. Overall, our findings suggest that NTCU induces pLSCC through an atypical bronchiolar hyperplasia-metaplasia-dysplasia-SCC sequence in mouse lung bronchioles. Notably, Ki67-positive deltaNp63(pos)CD44v(pos) cancer cells, cancer cells overexpressing phosphorylated epidermal growth factor receptor and signal transducer and activator of transcription 3, and tumor-associated macrophages were all present in far greater numbers in the peripheral area of the pLSCCs compared with the central area. These findings suggest that deltaNp63(pos)CD44v(pos) clara cells in mouse lung bronchioles might be the origin of the NTCU-induced pLSCCs. Our findings also suggest that tumor-associated macrophages may contribute to creating a tumor microenvironment in the peripheral area of pLSCCs that allows deltaNp63(pos)CD44v(pos) cancer cell expansion through activation of epidermal growth factor receptor signaling, and that exerts an immunosuppressive effect through activation of signal transducer and activator of transcription 3 signaling.

Examination of in vivo mutagenicity of sodium arsenite and dimethylarsinic acid in gpt delta rats.

Arsenic is a well-known human bladder and liver carcinogen, but its exact mechanism of carcinogenicity is not fully understood. Dimethylarsinic acid (DMAV) is a major urinary metabolite of sodium arsenite (iAsIII) and induces urinary bladder cancers in rats. DMAV and iAsIII are negative in in vitro mutagenicity tests. However, their in vivo mutagenicities have not been determined. The purpose of present study is to evaluate the in vivo mutagenicities of DMAV and iAsIII in rat urinary bladder epithelium and liver using gpt delta F344 rats. Ten-week old male gpt delta F344 rats were randomized into 3 groups and administered 0, 92mg/L DMAV, or 87mg/L iAsIII (each 50mg/L As) for 13weeks in the drinking water. In the mutation assay, point mutations are detected in the gpt gene by 6-thioguanine selection (gpt assay) and deletion mutations are identified in the red/gam genes by Spi- selection (Spi- assay). Results of the gpt and Spi- assays showed that DMAV and iAsIII had no effects on the mutant frequencies or mutation spectrum in urinary bladder epithelium or liver. These findings indicate that DMAV and iAsIII are not mutagenic in urinary bladder epithelium or liver in rats.

Targeted delivery of ubiquitin-conjugated BH3 peptide-based Mcl-1 inhibitors into cancer cells.

BH3 peptides are key mediators of apoptosis and have served as the lead structures for the development of anticancer therapeutics. Previously, we reported the application of a simple cysteine-based side chain cross-linking chemistry to NoxaBH3 peptides that led to the generation of the cross-linked NoxaBH3 peptides with increased cell permeability and higher inhibitory activity against Mcl-1 ( Muppidi, A., Doi, K., Edwardraja, S., Drake, E. J., Gulick, A. M., Wang, H.-G., Lin, Q. ( 2012 ) J. Am. Chem. Soc. 134 , 14734 ). To deliver cross-linked NoxaBH3 peptides selectively into cancer cells for enhanced efficacy and reduced systemic toxicity, here we report the conjugation of the NoxaBH3 peptides with the extracellular ubiquitin, a recently identified endogenous ligand for CXCR4, a chemokine receptor overexpressed in cancer cells. The resulting ubiquitin-NoxaBH3 peptide conjugates showed increased inhibitory activity against Mcl-1 and selective killing of the CXCR4-expressing cancer cells. The successful delivery of the NoxaBH3 peptides by ubiquitin into cancer cells suggests that the ubiquitin/CXCR4 axis may serve as a general route for the targeted delivery of anticancer agents.

Synthesis of cell-permeable stapled BH3 peptide-based Mcl-1 inhibitors containing simple aryl and vinylaryl cross-linkers.

We report the synthesis of a series of distance-matching aryl and vinylaryl cross-linkers for constructing stapled peptides containing cysteines at i,i+7 positions. Langevin dynamics simulation studies helped to classify these cross-linkers into two categories: the rigid cross-linkers with narrower S-S distance distribution and the flexible cross-linkers with wider S-S distance distribution. The stapled Noxa BH3 peptides with the flexible distance-matching cross-linkers gave the highest degree of helicity as well as the most potent inhibitory activity against Mcl-1. However, the stapled peptides with the highest hydrophobicity showed the most efficient cellular uptake. Together, this work illustrates the divergent nature of binding affinity and cellular uptake, and the vital importance of choosing appropriate cross-linkers in constructing stapled peptides with the drug-like properties.

Incidence and clinical outcome of lateral femoral cutaneous nerve injury after periacetabular osteotomy.

Aims: Lateral femoral cutaneous nerve (LFCN) injury is a complication after periacetabular osteo-tomy (PAO) using an anterior approach, which might adversely affect the outcome. However, no prospective study has assessed the incidence and severity of this injury and its effect on the clinical outcomes over a period of time for longer than one year after PAO. The aim of this study was to assess the incidence and severity of the symptoms of LFCN injury for ≥ three years after PAO and report its effect on clinical outcomes. Methods: A total of 40 hips in 40 consecutive patients who underwent PAO between May 2016 and July 2018 were included in the study, as further follow-up of the same patients from a previous study. We prospectively evaluated the incidence, severity, and area of symptoms following LFCN injury. We also recorded the clinical scores at one year and ≥ three years postoperatively using the 36-Item Short Form Health Survey (SF-36) and Japanese Orthopaedic Association Hip Disease Evaluation Questionnaire (JHEQ) scores. Results: A total of 20 patients (50%) had symptoms of a LFCN injury at one year after PAO. At ≥ three years postoperatively, the symptoms had completely resolved in seven of these patients and 13 (33%) had persistent symptoms. The severity and area of symptoms did not significantly differ between one and ≥ three years postoperatively. The JHEQ showed significant differences in the patient satisfaction and mental scores between those with and those without sypmtoms of LFCN injury at ≥ three years postoperatively, while there was no significant difference in the mean SF-36 scores. Conclusion: The incidence of LFCN injury after PAO using an anterior approach is high. The outcome of PAO, ≥ three years postoperatively, is poorer in patients with persistent symptoms from a perioperative LFCN injury, in that patient satisfaction and mental health scores are adversely affected.

Life Course Epidemiology of Hip Osteoarthritis in Japan: A Multicenter, Cross-Sectional Study.

BACKGROUND: The incidence of developmental dysplasia of the hip (DDH) in Japanese newborns has reduced drastically following a primary prevention campaign initiated around 1972 to 1973; this perinatal education campaign promoted maintaining the hips of newborns in the naturally flexed-leg position. The purpose of the present study was to describe the life course epidemiology of hip osteoarthritis (OA) in adolescent and adult patients and to assess its association with exposure to the primary prevention campaign for DDH. METHODS: We included new patients with hip OA diagnosed from January 1, 2022, to December 31, 2022, at 12 core hospitals (8 special-function hospitals and 4 regional medical care support hospitals). The trend in the percentage of hips with a history of DDH treatment in childhood was estimated with use of a centered moving average using the birth year of the patient. We compared the prevalence of severe subluxation (Crowe type II, III, or IV) between patients with secondary hip OA due to hip dysplasia who were born in or before 1972 and those who were born in or after 1973. RESULTS: Overall, 1,095 patients (1,381 hips) were included. The mean age at the time of the survey was 63.5 years (range, 15 to 95 years). A total of 795 patients (1,019 hips; 73.8% of hips) were diagnosed with secondary OA due to hip dysplasia. Approximately 13% to 15% of hips among patients born from 1963 to 1972 had a history of DDH treatment in childhood; however, the percentage decreased among patients born in or after 1973. The prevalence of severe subluxation (Crowe type II, III, or IV) among patients born in or after 1973 was 2.4%, which was significantly less than that among patients born in or before 1972 (11.1%; odds ratio, 0.20; p < 0.001). CONCLUSIONS: As of 2022, secondary hip OA due to hip dysplasia is still responsible for most new cases of adolescent and adult hip OA seen in core hospitals in Japan. However, the perinatal education campaign initiated 50 years ago, which utilized a population approach and advocated for maintaining the hips of newborns in the naturally flexed-leg position, may have improved the environmental factors of DDH, as indicated by the apparently reduced need for treatment of DDH in childhood and the associated severe subluxation. This may result in a reduced need for challenging hip surgery later in life. LEVEL OF EVIDENCE: Prognostic Level III . See Instructions for Authors for a complete description of levels of evidence.

Discovery of marinopyrrole A (maritoclax) as a selective Mcl-1 antagonist that overcomes ABT-737 resistance by binding to and targeting Mcl-1 for proteasomal degradation.

The anti-apoptotic Bcl-2 family of proteins, including Bcl-2, Bcl-X(L) and Mcl-1, are well-validated drug targets for cancer treatment. Several small molecules have been designed to interfere with Bcl-2 and its fellow pro-survival family members. While ABT-737 and its orally active analog ABT-263 are the most potent and specific inhibitors to date that bind Bcl-2 and Bcl-X(L) with high affinity but have a much lower affinity for Mcl-1, they are not very effective as single agents in certain cancer types because of elevated levels of Mcl-1. Accordingly, compounds that specifically target Mcl-1 may overcome this resistance. In this study, we identified and characterized the natural product marinopyrrole A as a novel Mcl-1-specific inhibitor and named it maritoclax. We found that maritoclax binds to Mcl-1, but not Bcl-X(L), and is able to disrupt the interaction between Bim and Mcl-1. Moreover, maritoclax induces Mcl-1 degradation via the proteasome system, which is associated with the pro-apoptotic activity of maritoclax. Importantly, maritoclax selectively kills Mcl-1-dependent, but not Bcl-2- or Bcl-X(L)-dependent, leukemia cells and markedly enhances the efficacy of ABT-737 against hematologic malignancies, including K562, Raji, and multidrug-resistant HL60/VCR, by ∼60- to 2000-fold at 1-2 µM. Taken together, these results suggest that maritoclax represents a new class of Mcl-1 inhibitors, which antagonizes Mcl-1 and overcomes ABT-737 resistance by targeting Mcl-1 for degradation.

Clinical Outcomes for Total Hip Arthroplasty with and without Previous Curved Periacetabular Osteotomy.

There are currently no reports on the clinical outcomes after total hip arthroplasty (THA) with previous curved periacetabular osteotomy (CPO), although the outcomes after THA with non-CPO types of periacetabular osteotomy have been reported. This study aimed to clarify the differences in clinical outcomes and radiographic features after THA with or without previous CPO. We performed a retrospective case-control with individual matching study. The participants were 10 patients with 11 hips that underwent cementless THA between October 1998 and October 2018 with previous CPO (osteotomy group). For the control group, we matched age, sex, and follow-up period, and included 32 patients with 33 hips that underwent cementless THA without previous CPO at a 1:3 ratio. The Harris Hip Score (HHS), cup size, position, and alignment, global offset (GO), operative time, perioperative blood loss, frequency of osteophyte removal, and major complications were compared between the two groups. The osteotomy group had no cases with revision surgery and dislocation. No significant differences were found between the two groups as follows: mean HHS, 94.9 points in the osteotomy group versus 92.7 points in the control group at the final follow-up; mean GO, 70.1 mm in the osteotomy group versus 71.4 mm in the control group; cup size, position, and alignment after THA; operative time; and perioperative blood loss. The frequency of osteophyte removal was higher in the osteotomy group. The take-home messages were that the clinical outcomes, including HHS, and radiographic features, including GO, after THA were equivalent in the two groups.

A case of total hip arthroplasty for nonunion after femoral trochanteric fracture surgery with complete breakage of only the compression screw of the InterTAN nail.

Intramedullary nails are an effective treatment for common femoral trochanteric fractures. However, one of their complications is implant breakage due to poor reduction and nonunion after surgery. We herein report a case of a 54-year-old man who underwent total hip arthroplasty for nonunion after internal fixation of a femoral trochanteric fracture. The femoral trochanteric fracture was treated by internal fixation using the Trigen InterTAN nail. The patient developed symptoms of hip pain 6 months after internal fixation. Nine months after internal fixation, hip radiographs and computed tomography scans showed breakage of only the compression screw. During total hip arthroplasty, we were unable to remove the lag screw and compression screw before the femoral head dislocation because no gap was present between the two screws. Thus, we removed these screws with the femoral head after dislocation of the femoral head. The removed nail was partially damaged at the lag screw hole. This change was retrospectively observed on the preoperative computed tomography scan. Three months after total hip arthroplasty, the patient was able to walk unaided and the hip pain had resolved. If only the compression screw is completely broken after internal fixation with the Trigen InterTAN nail, both the lag screw and compression screw will be difficult to remove with preservation of the femoral head. We effectively managed such a case by not only revision internal fixation but also total hip arthroplasty.

Going beyond Binary: Rapid Identification of Protein-Protein Interaction Modulators Using a Multifragment Kinetic Target-Guided Synthesis Approach.

Kinetic target-guided synthesis (KTGS) is a powerful screening approach that enables identification of small molecule modulators for biomolecules. While many KTGS variants have emerged, a majority of the examples suffer from limited throughput and a poor signal/noise ratio, hampering reliable hit detection. Herein, we present our optimized multifragment KTGS screening strategy that tackles these limitations. This approach utilizes selected reaction monitoring liquid chromatography tandem mass spectrometry for hit detection, enabling the incubation of 190 fragment combinations per screening well. Consequentially, our fragment library was expanded from 81 possible combinations to 1710, representing the largest KTGS screening library assembled to date. The expanded library was screened against Mcl-1, leading to the discovery of 24 inhibitors. This work unveils the true potential of KTGS with respect to the rapid and reliable identification of hits, further highlighting its utility as a complement to the existing repertoire of screening methods used in drug discovery.

The BH3 alpha-helical mimic BH3-M6 disrupts Bcl-X(L), Bcl-2, and MCL-1 protein-protein interactions with Bax, Bak, Bad, or Bim and induces apoptosis in a Bax- and Bim-dependent manner.

A critical hallmark of cancer cell survival is evasion of apoptosis. This is commonly due to overexpression of anti-apoptotic proteins such as Bcl-2, Bcl-X(L), and Mcl-1, which bind to the BH3 α-helical domain of pro-apoptotic proteins such as Bax, Bak, Bad, and Bim, and inhibit their function. We designed a BH3 α-helical mimetic BH3-M6 that binds to Bcl-X(L) and Mcl-1 and prevents their binding to fluorescently labeled Bak- or Bim-BH3 peptides in vitro. Using several approaches, we demonstrate that BH3-M6 is a pan-Bcl-2 antagonist that inhibits the binding of Bcl-X(L), Bcl-2, and Mcl-1 to multi-domain Bax or Bak, or BH3-only Bim or Bad in cell-free systems and in intact human cancer cells, freeing up pro-apoptotic proteins to induce apoptosis. BH3-M6 disruption of these protein-protein interactions is associated with cytochrome c release from mitochondria, caspase-3 activation and PARP cleavage. Using caspase inhibitors and Bax and Bak siRNAs, we demonstrate that BH3-M6-induced apoptosis is caspase- and Bax-, but not Bak-dependent. Furthermore, BH3-M6 disrupts Bcl-X(L)/Bim, Bcl-2/Bim, and Mcl-1/Bim protein-protein interactions and frees up Bim to induce apoptosis in human cancer cells that depend for tumor survival on the neutralization of Bim with Bcl-X(L), Bcl-2, or Mcl-1. Finally, BH3-M6 sensitizes cells to apoptosis induced by the proteasome inhibitor CEP-1612.

Rational design of proteolytically stable, cell-permeable peptide-based selective Mcl-1 inhibitors.

Direct chemical modifications provide a simple and effective means to "translate" bioactive helical peptides into potential therapeutics targeting intracellular protein-protein interactions. We previously showed that distance-matching bisaryl cross-linkers can reinforce peptide helices containing two cysteines at the i and i+7 positions and confer cell permeability to the cross-linked peptides. Here we report the first crystal structure of a biphenyl-cross-linked Noxa peptide in complex with its target Mcl-1 at 2.0 Å resolution. Guided by this structure, we remodeled the surface of this cross-linked peptide through side-chain substitution and N-methylation and obtained a pair of cross-linked peptides with substantially increased helicity, cell permeability, proteolytic stability, and cell-killing activity in Mcl-1-overexpressing U937 cells.

Synthesis and evaluation of substituted hexahydronaphthalenes as novel inhibitors of the Mcl-1/BimBH3 interaction.

Mcl-1, an anti-apoptotic member of the Bcl-2 protein family, is overexpressed in a broad range of human cancers and plays a critical role in conferring resistance to chemotherapy. In the course of screening a natural product-like library of sesquiterpenoid analogs, we identified substituted hexahydronaphthalenes that showed activity against the Mcl-1/BimBH3 interaction in vitro. Here, we describe the synthesis of a small library of analogs and their biological evaluation. The most potent inhibitor in the series (19) exhibits an IC(50) of 8.3 µM by ELISA and disrupts the interaction between endogenously expressed Mcl-1 and Bim in cultured MDA-MB-468 breast cancer cells.

Enhancement of preneoplastic lesion yield by Chios Mastic Gum in a rat liver medium-term carcinogenesis bioassay.

The mastic (Pistacia lentiscus var. chia) tree is native throughout the Mediterranean region and has long proved a source of food additives and medical treatments. To investigate the modifying effects of Chios Mastic Gum on rat liver carcinogenesis, 6-week-old male F344 rats were subjected to the established rat liver medium-term carcinogenesis bioassay (Ito-test). At the commencement, rats (groups 1-4) were intraperitoneally injected with 200 mg/kg body weight of diethylnitrosamine (DEN). After two weeks, mastic was added to CRF (Charles River Formula)-1 powdered basal diet at doses of 0, 0.01, 0.1 and 1% in groups 1-4, respectively. At week 3, all rats were underwent two-thirds partial hepatectomy. The experiment was terminated at week 8. As results show, liver weights were significantly increased in a mastic dose-dependent manner among groups 1-4. The numbers (/cm(2)) and the areas (mm(2)/cm(2)) of glutathione S-transferase placental form (GST-P)-positive cell foci (>or=0.2 mm in diameter) were significantly increased in the DEN-1% group compared to the DEN-alone group, along with the average areas per foci and larger-sized foci (>or=0.4 mm). 5-Bromo-2'-deoxyuridine (BrdU)+GST-P double-immunohistochemistry showed the highest BrdU-labeling indices within GST-P foci in the DEN-1% group. 8-hydroxydeoxyguanosine (8-OHdG) levels in liver DNA did not vary, while real-time quantitative polymerase chain reaction (PCR) analysis of livers revealed many up- or down-regulated genes in the DEN-1% group. In conclusion, this is the first report to display a promotion potential of Chios Mastic Gum on the formation of preneoplastic lesions in the established rat liver medium-term carcinogenesis bioassay.