Basic accuracy of a 1D NOESY with presaturation method using standard solutions of amino and maleic acids.

1D NOESY with presaturation (NOESY-presat) is the most popular water suppression method. When D2O solutions of L-phenylalanine or L-valine were measured using NOESY, the absolute concentration biases increased with longer mixing and evolution times, reaching a maximum of 54% with respect to the preparation values. At mixing and evolution times of 0 ms and 0 µs, respectively, the absolute concentration biases were reduced to less than 3%. The remaining biases were caused by the off-resonance effect, which was prevented by setting the frequency offset to an intermediate value between the analyte and internal standard 3-(trimethylsilyl)-1-propanesulfonic acid-d6 (DSS-d6) signals. Nevertheless, NOESY-presat gave maximum absolute biases of 26% and 11% for glycine and maleic acid concentrations, respectively, in three H2O/D2O (90/10 vol%) solutions. The proposed NOESY-dual-presat method reduced the absolute biases to below 4%. However, water suppression was insufficient but was improved by setting the frequency offset to the same as the presaturation offset with the H2O signal, although the absolute biases rose to 5 to 13%. Quantitative analyses using NOESY-presat and NOESY-dual-presat require careful consideration of the off-resonance effect.

Expert-Novice Level Classification Using Graph Convolutional Network Introducing Confidence-Aware Node-Level Attention Mechanism.

In this study, we propose a classification method of expert-novice levels using a graph convolutional network (GCN) with a confidence-aware node-level attention mechanism. In classification using an attention mechanism, highlighted features may not be significant for accurate classification, thereby degrading classification performance. To address this issue, the proposed method introduces a confidence-aware node-level attention mechanism into a spatiotemporal attention GCN (STA-GCN) for the classification of expert-novice levels. Consequently, our method can contrast the attention value of each node on the basis of the confidence measure of the classification, which solves the problem of classification approaches using attention mechanisms and realizes accurate classification. Furthermore, because the expert-novice levels have ordinalities, using a classification model that considers ordinalities improves the classification performance. The proposed method involves a model that minimizes a loss function that considers the ordinalities of classes to be classified. By implementing the above approaches, the expert-novice level classification performance is improved.

Fresh Dual Presaturation Method for Analyzing H2O-Rich Samples Using Quantitative 1H NMR.

A fresh dual presaturation (pre-SAT) method was developed to quantify analytes accurately near the suppressed water signal in 1H NMR spectra obtained from H2O-rich samples. The method includes an extra dummy pre-SAT with a suitable offset for each analyte signal in addition to the water pre-SAT. The residual HOD signal at 4.66 ppm was observed using D2O solutions containing l-phenylalanine (Phe) or l-valine (Val) and an internal standard of 3-(trimethylsilyl)-1-propanesulfonic acid-d6 sodium salt (DSS-d6). When the HOD signal was suppressed using the conventional single pre-SAT method, the measured concentration of Phe from the NCH signal at 3.89 ppm decreased by a maximum of 48%, whereas the dual pre-SAT method gave a reduction in the Phe concentration measured from the NCH signal of less than 3%. The proposed dual pre-SAT method achieved accurate quantification of glycine (Gly) and maleic acid (MA) in a 10 vol % D2O/H2O solution. The measured concentrations of Gly of 513.5 ± 8.9 mg kg-1 and MA of 512.2 ± 10.3 mg kg-1 corresponded to sample preparation values of Gly of 502.9 ± 1.7 mg kg-1 and MA of 506.7 ± 2.9 mg kg-1 (the number after "±" indicates the expanded uncertainty (k = 2)).

Macrolide and Ketolide Antibiotics Inhibit the Cytotoxic Effect of Trastuzumab Emtansine in HER2-Positive Breast Cancer Cells: Implication of a Potential Drug-ADC Interaction in Cancer Chemotherapy.

Macrolides are widely used for the long-term treatment of infections and chronic inflammatory diseases. The pharmacokinetic features of macrolides include extensive tissue distribution because of favorable membrane permeability and accumulation within lysosomes. Trastuzumab emtansine (T-DM1), a HER2-targeting antibody-drug conjugate (ADC), is catabolized in the lysosomes, where Lys-SMCC-DM1, a potent cytotoxic agent, is processed by proteinase degradation and subsequently released from the lysosomes to the cytoplasm through the lysosomal membrane transporter SLC46A3, resulting in an antitumor effect. We recently demonstrated that erythromycin and clarithromycin inhibit SLC46A3 and attenuate the cytotoxicity of T-DM1; however, the effect of other macrolides and ketolides has not been determined. In this study, we evaluated the effect of macrolide and ketolide antibiotics on T-DM1 cytotoxicity in a human breast cancer cell line, KPL-4. Macrolides used in the clinic, such as roxithromycin, azithromycin, and josamycin, as well as solithromycin, a ketolide under clinical development, significantly attenuated T-DM1 cytotoxicity in addition to erythromycin and clarithromycin. Of these, azithromycin was the most potent inhibitor of T-DM1 efficacy. These antibiotics significantly inhibited the transport function of SLC46A3 in a concentration-dependent manner. Moreover, these compounds extensively accumulated in the lysosomes at the levels estimated to be 0.41-13.6 mM when cells were incubated with them at a 2 µM concentration. The immunofluorescence staining of trastuzumab revealed that azithromycin and solithromycin inhibit the degradation of T-DM1 in the lysosomes. These results suggest that the attenuation of T-DM1 cytotoxicity by macrolide and ketolide antibiotics involves their lysosomal accumulation and results in their greater lysosomal concentrations to inhibit the SLC46A3 function and T-DM1 degradation. This suggests a potential drug-ADC interaction during cancer chemotherapy.

Simple Strategy for Benzo[g]chromene Synthesis via a Photochemical Intramolecular Cyclization Reaction.

Photochemical reactions are often a desirable strategy for organic synthesis because they do not require toxic and expensive reagents and produce less waste than thermal reactions. Herein, a facile photochemical strategy is described to synthesize benzo[g]chromene derivatives. This strategy utilizes the photoredox reaction of quinones, which allows C-H bond oxidation in the vicinity of the photoexcited quinone carbonyl group. The reaction mechanism was investigated using 1H NMR analysis. The intramolecular cyclization reaction proceeded via the formation of 1,3-dioxole compounds as intermediates by the photoredox reaction of p-quinone, followed by re-cyclization. The synthesized benzo[g]chromene derivatives are important heterocyclic skeletons with useful biological and pharmacological properties.

Transformation of Renieramycin M into Renieramycins T and S by Intramolecular Photoredox Reaction of 7-Methoxy-6-methyl-1,2,3,4-tetrahydroisoquinoline-5,8-dione Derivatives.

In connection with our studies of biologically active 1,2,3,4-tetrahydroisoquinoline marine natural products, we describe herein a useful intramolecular photoredox transformation of 7-methoxy-6-methyl-1,2,3,4-tetrahydroisoquinoline-5,8-dione tricyclic models into 5-hydroxy-tetrahydroisoquinol[1,3]dioxoles in excellent yields. We applied this methodology to the transformation of renieramycin M into renieramycins T and S and the transformation of saframycin A. The results of cytotoxicity studies are also presented.

Light-Mediated Transformation of Renieramycins and Semisynthesis of 4'-Pyridinecarbonyl-Substituted Renieramycin-Type Derivatives as Potential Cytotoxic Agents against Non-Small-Cell Lung Cancer Cells.

The semisynthesis of renieramycin-type derivatives was achieved under mild and facile conditions by attaching a 1,3-dioxole-bridged phenolic moiety onto ring A of the renieramycin structure and adding a 4'-pyridinecarbonyl ester substituent at its C-5 or C-22 position. These were accomplished through a light-induced intramolecular photoredox reaction using blue light (4 W) and Steglich esterification, respectively. Renieramycin M (4), a bis-tetrahydroisoquinolinequinone compound isolated from the Thai blue sponge (Xestospongia sp.), served as the starting material. The cytotoxicity of the 10 natural and semisynthesized renieramycins against non-small-cell lung cancer (NSCLC) cell lines was evaluated. The 5-O-(4'-pyridinecarbonyl) renieramycin T (11) compound exhibited high cytotoxicity with half-maximal inhibitory concentration (IC50) values of 35.27 ± 1.09 and 34.77 ± 2.19 nM against H290 and H460 cells, respectively. Notably, the potency of compound 11 was 2-fold more than that of renieramycin T (7) and equal to those of 4 and doxorubicin. Interestingly, the renieramycin-type derivatives with a hydroxyl group at C-5 and C-22 exhibited weak cytotoxicity. In silico molecular docking and dynamics studies confirmed that the mitogen-activated proteins, kinase 1 and 3 (MAPK1 and MAPK3), are suitable targets for 11. Thus, the structure-cytotoxicity study of renieramycins was extended to facilitate the development of potential anticancer agents for NSCLC cells.

Skin quality sensor to evaluate vibration and friction generated when sliding over skins.

OBJECTIVE: The mechanical condition and tactile evaluation of skin are essential for the development of skin care products. Most of the existing commercial instruments and studies aim to evaluate the skin surface by pressing it for hardness or by using imaging sensors, but there have been few instrumental measurements employing rubbing motion. Here, we have developed a sensor specialized for tactile sensation and the contact phenomenon during skin rubbing. METHODS: The developed sensor has three features: It can measure body parts including cheeks and arms, automate the rubbing motion of the probe and measure vibration and friction simultaneously. It is hand-held, with metal probes that rub the skin surface while rotating under a motor drive; it has an accelerometer and a force sensor beneath the probe measuring vibration and friction forces. To evaluate the validity of the sensor's measurements, artificial skin models were measured using the developed sensor and commercially available sensors and the results were compared. The relationship between the sensor output, surface roughness measurement and sensory evaluation was also investigated. Additionally, we evaluated the inter-rater reliability when measuring actual skin. RESULTS: The measurements of five artificial skin models with different surface shapes showed a high correlation (r = 0.99) between the vibration intensity values evaluated by the developed sensor and those measured by a tri-axial acceleration sensor attached to a fingernail. The correlation coefficient between the vibration intensity values and surface roughness was r = 0.91, and the correlation with the sensory evaluation score of roughness was r = 0.99. The friction coefficients measured by the developed sensor and the force plate had r = 0.93, based on measurements of five artificial skin models with different friction conditions. The inter-rater correlation coefficients between the three participants of the developed sensor were as high as 0.92 and 0.94 for the vibration and friction measurements respectively. CONCLUSION: The vibration intensities and friction coefficients from the sensor were highly correlated with those of the conventional sensor. The inter-rater reliability was also high. The developed sensor can be useful for tactile evaluation in skin-care product development.

OBJECTIF: l'état mécanique et l'évaluation tactile de la peau sont essentiels au développement de produits de soins de la peau. La plupart des instruments disponibles sur le marché et des études publiées à ce jour évaluent la surface de la peau en la comprimant pour déterminer sa dureté ou en utilisant des capteurs d'imagerie, mais il n'y a eu que peu de mesures instrumentales utilisant le mouvement de frottement. Ici, nous avons développé un capteur spécialisé pour la sensation tactile et le phénomène de contact lors du frottement de la peau. MÉTHODES: le capteur développé possède trois caractéristiques : il permet d'exercer des mesures sur plusieurs parties du corps, y compris les joues et les bras ; il automatise le mouvement de frottement de la sonde et il mesure simultanément les vibrations et les frottements. Tenu à la main, doté de sondes en métal qui frottent la surface de la peau tout en tournant sous l'action d'un moteur, il est équipé d'un accéléromètre et d'un capteur de force situé sous la sonde qui mesure les forces de vibration et de frottement. Pour déterminer la validité des mesures du capteur, des modèles de peau artificielles ont été évalués à l'aide du capteur développé et de capteurs déjà disponibles sur le marché, et les résultats ont été comparés. Le lien entre les mesures réalisées à l'aide du capteur, la mesure de la rugosité de la surface de la peau et l'évaluation sensorielle a également été étudié. En outre, nous avons évalué la fiabilité inter-évaluateurs lors de la mesure réelle de la peau. RÉSULTATS: les mesures de cinq modèles de peau artificielle avec des formes de surface différentes ont montré une forte corrélation (r = 0,99) entre les valeurs d'intensité des vibrations évaluées par le capteur développé et celles mesurées par un capteur d'accélération triaxial fixé à un ongle. Le coefficient de corrélation entre les valeurs d'intensité des vibrations et la rugosité de la surface était r = 0,91, et la corrélation avec le score d'évaluation sensorielle de la rugosité était r = 0,99. Les coefficients de frottement mesurés par le capteur développé et la plaque de force étaient r = 0,93, sur la base des mesures de cinq modèles de peau artificielle avec des conditions de frottement différentes. Les coefficients de corrélation inter-évaluateurs entre les trois participants utilisant le capteur développé ont atteint 0,92 et 0,94 pour les mesures de vibrations et de frottement, respectivement. CONCLUSION: les intensités des vibrations et les coefficients de frottement du capteur se sont avérés fortement corrélés avec ceux du capteur conventionnel. La fiabilité inter-évaluateurs était également élevée. Le capteur développé peut être utile pour l'évaluation tactile lors du développement de produits de soins de la peau.

[Right Atrial Blood Cyst with Patent Foramen Ovale:Report of a Case].

A woman in 70s was diagnosed with lung cancer, and a right atrial mass was discovered incidentally during preoperative examination by contrast-enhanced computed tomography (CT). Transesophageal echocardiography revealed a 20-mm, stemmed, spherical mass with low internal echogenicity and partially high echogenicity extending from the junction of the inferior vena cava to the posterior wall of the right atrium. Patent foramen ovale( PFO) was also confirmed. To avoid embolization and obtain diagnosis, the patient was referred for right atrial tumor resection. Cardiopulmonary bypass was established; the right atrial tumor was removed while the patient was in cardiac arrest. The tumor membrane was thin and easily ruptured, revealing jelly-like blood content and calcified mass. The patient recovered well after surgery and was discharged on day 15. According to the pathological examination, the tumor was a blood cyst. This is an extremely rare case of a blood cyst with PFO.

Smart Pneumatic Artificial Muscle Using a Bend Sensor like a Human Muscle with a Muscle Spindle.

Shortage of labor and increased work of young people are causing problems in terms of care and welfare of a growing proportion of elderly people. This is a looming social problem because people of advanced ages are increasing. Necessary in the fields of care and welfare, pneumatic artificial muscles in actuators of robots are being examined. Pneumatic artificial muscles have a high output per unit of weight, and they are soft, similarly to human muscles. However, in previous research of robots using pneumatic artificial muscles, rigid sensors were often installed at joints and other locations due to the robots' structures. Therefore, we developed a smart actuator that integrates a bending sensor that functions as a human muscle spindle; it can be externally attached to the pneumatic artificial muscle. This paper reports a smart artificial muscle actuator that can sense contraction, which can be applied to developed self-monitoring and robot posture control.

[Racemose Hemangioma of the Bronchial Artery with Pulmonary Artery Fistula].

A 55-year-old woman was suspected of having hilar lymph node enlargement on a routine examination of the chest computed tomography( CT) scan at our hospital. On further examination, thoracic contrast CT and bronchial arteriography showed prominent dilation and meandering of the right bronchial artery with an aneurysm which formed a fistula to the pulmonary artery A7 at its distal end. Diagnosed as racemose hemangioma of bronchial artery with pulmonary artery fistula, we performed a surgical resection by open thoracotomy on the second day following bronchial artery embolization( BAE). No recurrence was observed in the five years of follow-up period after surgery. Therefore, surgical resection for asymptomatic racemose hemangioma of bronchial artery with bronchial artery-pulmonary artery fistula can be considered an effective method for the long-term prevention of bleeding.

Chemical Research on Antitumor Isoquinoline Marine Natural Products and Related Compounds.

The biologically active, naturally occurring 1,2,3,4-tetrahydroisoquinoline-quinone (THIQ) family members isolated from Actinomycetes and marine organisms have been studied thoroughly over the past five decades. Among them, marine natural products along with their reduced compounds, such as renieramycins and ecteinascidins, have attracted interest due to their fantastic structures and meager availability in nature as well as their potent antitumor profiles. As part of our search for new anticancer metabolites through the isolation and characterization of anticancer THIQ compounds from Thai marine animals, we have developed a fascinating THIQ natural product chemistry and medicinal chemistry based on knowledge of the chemistry of saframycin antibiotics as well as their isolation, characterization, transformation, partial synthesis, and total synthesis. This review mainly presents our contributions during 1999-2019 to the field of research on biologically active renieramycin along with ecteinascidin marine natural products.

Rubber Material Property Prediction Using Electron Microscope Images of Internal Structures Taken under Multiple Conditions.

A method for prediction of properties of rubber materials utilizing electron microscope images of internal structures taken under multiple conditions is presented in this paper. Electron microscope images of rubber materials are taken under several conditions, and effective conditions for the prediction of properties are different for each rubber material. Novel approaches for the selection and integration of reliable prediction results are used in the proposed method. The proposed method enables selection of reliable results based on prediction intervals that can be derived by the predictors that are each constructed from electron microscope images taken under each condition. By monitoring the relationship between prediction results and prediction intervals derived from the corresponding predictors, it can be determined whether the target prediction results are reliable. Furthermore, the proposed method integrates the selected reliable results based on Dempster-Shafer (DS) evidence theory, and this integration result is regarded as a final prediction result. The DS evidence theory enables integration of multiple prediction results, even if the results are obtained from different imaging conditions. This means that integration can even be realized if electron microscope images of each material are taken under different conditions and even if these conditions are different for target materials. This nonconventional approach is suitable for our application, i.e., property prediction. Experiments on rubber material data showed that the evaluation index mean absolute percent error (MAPE) was under 10% by the proposed method. The performance of the proposed method outperformed conventional comparative property estimation methods. Consequently, the proposed method can realize accurate prediction of the properties with consideration of the characteristic of electron microscope images described above.

[Endovascular Therapy for Kinking of an Open Stent Graft after Stanford Type A Acute Aortic Dissection Repair:Report of a Case].

This is the case of 50s female with Stanford type A acute aortic dissection who underwent emergent total arch replacement. The aortic arch was transected just distal to the left subclavian artery, followed by the insertion of J Graft Frozenix into the descending aorta. No blood pressure gradient was observed between the radial and femoral arteries immediately after the operation. Nevertheless, intermittent claudication was observed after a week. Ankle-brachial index( ABI) measurement was calculated at 0.7 in both legs. Computed tomography (CT) revealed a kinking of the non-stented part of the endograft. Subsequently, thoracic endovascular aortic repair( TEVAR) was performed. As a result, ABI measurement normalized and lower limb pain disappeared. Three years after, CT showed that the endograft expanded satisfactorily. In deployment of J Graft Frozenix, the non-stented part should be kept as short as possible. For kinking, TEVAR should be considered the initial treatment option.

Unique Usage of a Classical Selective Homodecoupling Sequence for High-Resolution Quantitative 1H NMR.

Classical selective homodecoupling was used in a 1H NMR purity assay to improve accuracy by overcoming spectral overlaps due to 1H-1H spin coupling. Dummy irradiation at a specific frequency was used in addition to irradiation at a 1H resonance of the analyte to avoid irradiation bias. The method was validated in a 1H NMR purity assay of high-purity diethyl phthalate (National Metrology Institute of Japan Certified Reference Material (NMIJ CRM), purity: 99.98%). The obtained purity value biases were 0.27% or less. The utility of the method was demonstrated in another 1H NMR purity assay of dipropyl phthalate (NMIJ CRM, purity: 98.41%), which contained a tiny amount of the structurally similar compound methyl propyl phthalate as an impurity. An accurate assay was achieved with the method, giving a purity of 98.39%, whereas the conventional method gave a purity 99.13%.

Postmortem CT and MRI findings of massive fat embolism.

OBJECTIVE: To elucidate postmortem computed tomography (PMCT) and postmortem magnetic resonance (PMMR) imaging findings suggesting massive fat embolism. MATERIALS AND METHODS: Consecutive forensic cases with PMCT and PMMR scans of subjects prior to autopsy were assessed. For PMCT, 16- or 64-row multidetector CT scans were used; for PMMR, a 1.5 T system was used. MRI sequences of the chest area included T2- and T1-weighted fast spin-echo imaging, T2*-weighted imaging, T1-weighted 3-dimensional gradient-echo imaging with or without a fat-suppression pulse, short tau inversion recovery, and in-phase/opposed-phase imaging. At autopsy, forensic pathologists checked for pulmonary fat embolism with fat staining; Falzi's grading system was used for classification. RESULTS: Of 31 subjects, four were excluded because fat staining for histopathological examination of the lung tissue could not be performed. In three of the remaining 27 subjects, histology revealed massive fat embolism (Falzi grade III) and the cause of death was considered to be associated with fat embolism. CT detected a "fat-fluid level" in the right heart or intraluminal fat in the pulmonary arterial branches in two subjects. MRI detected these findings more clearly in both subjects. In one subject, CT and MRI were both negative. There were no positive findings in the 24 subjects that were fat embolism-negative by histology. DISCUSSION AND CONCLUSION: In some subjects, a massive fat embolism can be suggested by postmortem imaging with a "fat-fluid level" in the right heart or intraluminal fat in the pulmonary arterial branches. PMMR potentially suggests fat embolism more clearly than PMCT.

Chemistry of Renieramycins. Part 19: Semi-Syntheses of 22-O-Amino Ester and Hydroquinone 5-O-Amino Ester Derivatives of Renieramycin M and Their Cytotoxicity against Non-Small-Cell Lung Cancer Cell Lines.

Two new series of synthetic renieramycins including 22-O-amino ester and hydroquinone 5-O-amino ester derivatives of renieramycin M were semi-synthesized and evaluated for their cytotoxicity against the metastatic non-small-cell lung cancer H292 and H460 cell lines. Interestingly, the series of 22-O-amino ester derivatives displayed a potent cytotoxic activity greater than the hydroquinone derivatives. The most cytotoxic derivative of the series was the 22-O-(N-Boc-l-glycine) ester of renieramycin M (5a: IC50 3.56 nM), which showed 7-fold higher potency than renieramycin M (IC50 24.56 nM) and 61-fold more than jorunnamycin A (IC50 217.43 nM) against H292 cells. In addition, 5a exhibited a significantly higher cytotoxic activity than doxorubicin (ca. 100 times). The new semi-synthetic renieramycin derivatives will be further studied and developed as potential cytotoxic agents for non-small-cell lung cancer treatment.

Naturally occurring biflavonoids with amyloid ß aggregation inhibitory activity for development of anti-Alzheimer agents.

Amyloid ß (Aß) aggregation plays an essential role in promoting the progression of Alzheimer's disease (AD). Therefore, the inhibition of Aß aggregation is a potential therapeutic approach for AD. Herein, twenty-seven biflavonoids with different inter-flavonyl linkages and methoxy substitution patterns were isolated from several plants, and their Aß40 aggregation inhibitory activity was evaluated by the thioflavin-T fluorescence assay. Amentoflavone (1) and its monomethoxy derivatives (2, 3, and 5) exhibited the most potent inhibitory activity, with IC50 values of approximately 5 µM. It was clarified that increasing the number of methoxy substituents on the biflavonoid structures attenuated the inhibitory activity. Moreover, the linkage and the methoxy substitution pattern had a marked influence on the inhibitory activity. Our investigation strongly supports that biflavonoids can be considered a new type of anti-Alzheimer agents that may be successfully developed for AD patients.

5-O-Acetyl-Renieramycin T from Blue Sponge Xestospongia sp. Induces Lung Cancer Stem Cell Apoptosis.

Lung cancer is one of the most significant cancers as it accounts for almost 1 in 5 cancer deaths worldwide, with an increasing incident rate. Management of the cancer has been shown to frequently fail due to the ability of the cancer cells to resist therapy as well as metastasis. Recent evidence has suggested that the poor response to the current treatment drugs and the ability to undergo metastasis are driven by cancer stem cells (CSCs) within the tumor. The discovery of novel compounds able to suppress CSCs and sensitize the chemotherapeutic response could be beneficial to the improvement of clinical outcomes. Herein, we report for the first time that 5-O-acetyl-renieramycin T isolated from the blue sponge Xestospongia sp. mediated lung cancer cell death via the induction of p53-dependent apoptosis. Importantly, 5-O-acetyl-renieramycin T induced the death of CSCs as represented by the CSC markers CD44 and CD133, while the stem cell transcription factor Nanog was also found to be dramatically decreased in 5-O-acetyl-renieramycin T-treated cells. We also found that such a CSC suppression was due to the ability of the compound to deplete the protein kinase B (AKT) signal. Furthermore, 5-O-acetyl-renieramycin T was able to significantly sensitize cisplatin-mediated apoptosis in the lung cancer cells. Together, the present research findings indicate that this promising compound from the marine sponge is a potential candidate for anti-cancer approaches.

Renieramycin T Induces Lung Cancer Cell Apoptosis by Targeting Mcl-1 Degradation: A New Insight in the Mechanism of Action.

Among malignancies, lung cancer is the major cause of cancer death. Despite the advance in lung cancer therapy, the five-year survival rate is extremely restricted due to therapeutic failure and disease relapse. Targeted therapies selectively inhibiting certain molecules in cancer cells have been accepted as promising ways to control cancer. In lung cancer, evidence has suggested that the myeloid cell leukemia 1 (Mcl-1) protein, an anti-apoptotic member of the Bcl-2 family, is a target for drug action. Herein, we report the Mcl-1 targeting activity of renieramycin T (RT), a marine-derived tetrahydroisoquinoline alkaloid that was isolated from the Thai blue sponge Xestospongia sp. RT was shown to be dominantly toxic to lung cancer cells compared to the normal cells in the lung. The cytotoxicity of this compound toward lung cancer cells was mainly exerted through apoptosis induction. For the mechanism of action, we found that RT mediated activation of p53 protein and caspase-9 and -3 activations. While others Bcl-2 family proteins (Bcl-2, Bak, and Bax) were minimally changed in response to RT, Mcl-1 protein was dramatically diminished. We further performed the cycloheximide experiment and found that the half-life of Mcl-1 was significantly shortened by RT treatment. When MG132, a potent selective proteasome inhibitor, was utilized, it could restore the Mcl-1 level. Furthermore, immunoprecipitation analysis revealed that RT significantly increased the formation of Mcl-1-ubiquitin complex compared to the non-treated control. In conclusion, we report the potential apoptosis induction of RT with a mechanism of action involving the targeting of Mcl-1 for ubiquitin-proteasomal degradation. As Mcl-1 is critical for cancer cell survival and chemotherapeutic failure, this novel information regarding the Mcl-1-targeted compound would be beneficial for the development of efficient anti-cancer strategies or targeted therapies.