Anti-Leukemic Effect of Retinoic Acid in Non-APL AML Cells

Abstract [Background] Differentiation therapy with retinoic acid (RA) has remarkably improved the cure rate for acute promyelocytic leukemia (APL). RAs are known to have anti-leukemic effect in parts of non-APL acute myelocytic leukemia (AML) cells. It has been reported that AML patient with NPM1 or IDH1/2 mutation may have sensitivity to all-trans retinoic acid (ATRA) and the clinical study with combination of chemotherapy and ATRA is conducted in NPM1 mutated AML. In addition, ATRA and a FLT3 inhibitor, quizartinib, showed synergistic activity against FLT3-ITD positive AML patients' samples and their combination therapy improved the survival of AML xenograft mouse model. Moreover, it is also reported that the presence of super-enhancer at RARA gene locus is related to high RARA mRNA expression and sensitivity to tamibarotene. Another study showed that ATRA promote cancer apoptosis in the presence of Cellular Retinoic Acid-Binding Protein 1 (CRABP1) through G1 expansion. However, the efficacy of RAs and its mechanism in non-APL AML are still largely unknown. In this study, we evaluated anti-leukemic effect of RAs on primary AML cells and explored its predictive biomarkers for the purpose of optimization of therapy for non-APL AML. [Method] We examined growth inhibitory and differentiation effects of ATRA and tamibarotene on primary AML cells in vitro. Primary AML cells were purified by density-based centrifugation and cultured with various concentrations of RAs in methylcellulose-based media including cytokines. IC50 values of RAs were calculated by luminescent cell viable assay on day7 and 14. Cell differentiation upon RAs treatment was also examined in these cells by morphology and expression of CD11b using flow cytometry analysis. We also evaluated the efficacy of combination therapy of RAs and several tyrosine kinase inhibitors, quizartinib, midstaurin and dasatinib, and assessed their synergistic effect by calculating combination index. The association of IC50 values of RAs with clinical characteristics, mRNA expressions of RARA and CRABP1, and genetic mutations were investigated. [Result] We analyzed total of 30 primary non-APL AML samples and IC50 values of ATRA and tamibarotene were lower than 5 µM in 14 of 30 (46.7 %) and 4 of 14 (28.6 %) AML cells, respectively. There were no significant difference in IC50 value of ATRA and tamibarotene by FAB subtype and chromosomal abnormalities. ATRA and tamibarotene induced differentiation in 10 of the 22 and 2 of the 9, respectively, and anti-leukemic effect of ATRA and tamibarotene were not associated with these differentiation statuses. IC50 value of ATRA and tamibarotene were significantly lower in ASXL1 (p=0.0071 and p=0.0012, respectively), SRSF2 (p=0.0101 and p=0.0113, respectively), and IDH2 (p=0.0295 and p=0.0044, respectively) mutated AML cells. FLT3-ITD mutation with high allelic ratio (>0.5) was associated with high IC50 value of ATRA (p=0.0116). Although NPM1 mutation was associated with higher IC50 value of ATRA, all of 4 patients with NPM1 mutation concurrently have FLT3-ITD mutations in this study. Other gene mutations were not correlated with RAs sensitivity. The expression of RARA mRNA weakly correlated with sensitivity of ATRA and tamibarotene (P=0.1126 and P=0.0711 respectively), but it was not significant. [Conclusion] RAs were effective in almost half of non-APL AML cells in vitro and some AML cells were sensitive to ATRA without cell differentiation. Mutation in ASXL1, SRSF2, and IDH2 were identified as predictive factors for the RAs sensitivity, whereas the expression of RARA mRNA and other gene mutations were not correlated with RAs sensitivity. Disclosures Kiyoi: Takeda Pharmaceutical Co., Ltd.: Research Funding; Phizer Japan Inc.: Research Funding; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding; Chugai Pharmaceutical Co., Ltd.: Research Funding; FUJIFILM Corporation: Research Funding; Kyowa Hakko Kirin Co., Ltd.: Research Funding; Sanofi K.K.: Research Funding; Otsuka Pharmaceutical Co., Ltd.: Research Funding; Novartis Pharma K.K.: Research Funding; Zenyaku Kogyo Co., Ltd.: Research Funding; Eisai Co., Ltd.: Research Funding; Bristol-Myers Squibb: Honoraria; Celgene Corporation: Research Funding; Astellas Pharma Inc.: Research Funding; Nippon Shinyaku Co., Ltd.: Research Funding.

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Addition of All-Trans Retinoic Acid (ATRA) to the Combination of Fludarabine, Cytarabine, Idarubicin, with or without GCSF in Older, Higher Risk Patients with AML and High-Risk MDS Does Not Improve the Outcome in Those with NPM1 Mutation.

Blood ◽

10.1182/blood.v120.21.2616.2616 ◽

2012 ◽

Vol 120 (21) ◽

pp. 2616-2616 ◽

Cited By ~ 1

Author(s):

Aziz Nazha ◽

Carlos E. Bueso-Ramos ◽

Stefan Faderl ◽

Elihu H. Estey ◽

Susan M. O'Brien ◽

...

Keyword(s):

Retinoic Acid ◽

High Risk ◽

Research Funding ◽

Gene Mutations ◽

Npm1 Mutation ◽

Related Disease ◽

Trans Retinoic Acid ◽

All Trans Retinoic Acid ◽

Significant Difference ◽

The Impact

Abstract Abstract 2616 Background: Previous studies have been conflicting in their findings regarding whether NPM1 gene mutations may be a marker for response to all-trans retinoic acid (ATRA) given as an adjunct to intensive chemotherapy in patients with acute myeloid leukemia (AML) (Schlenk RF, Haematologica, 2009, 94, 54–60, Burnett AK, Blood, 2010, 115,948, and Schlenk RF, abstract #80, ASH 2011). Patients and Methods: we examined the impact of the addition of ATRA among patients with diploid cytogenetics treated on a randomized phase II study of fludarabine + cytarabine + idarubicine +/− GCSF +/− ATRA with available data on their NPM1 mutation status. Between September 1995 and November 1997, 215 patients with newly diagnosed AML (n=153) or high-risk MDS (RAEB or RAEB-T; n=62) were enrolled in the study. They had to have one of the unfavorable features of age > 71 years, antecedent hematological disorder (AHD), therapy- related disease, or high bilirubin (>2.9 mg/ml) or creatinine (>1.5 mg/ml) to be eligible to participate in the study (Estey E, Blood, 1999, 93, 2478). Among them 70 patients had diploid cytogenetic and are the subjects of this analysis. Results: The median age of the 70 patients was 66.5 (range, 23–87), 60% had AHD, 15.7% had therapy-related disease, and 40% were older than 71. Bone marrow samples were available for all patients for examination by immunohistochemistry (IHC) for the presence of cytoplasmic NPM (correlating to NPM1 mutation). Twenty (29%) of patients had NPM1 mutation. Among them 7 (35%) did and 13 (65%) did not receive ATRA in combination with chemotherapy. Complete remission (CR) was achieved in 5 (71.4%) of patients treated with ATRA as compared to 9 (69.2%) without ATRA (p=NS). The median event-free survival was 82 weeks vs. 56 weeks for patients receiving ATRA or not [(p=0.5), ranges, (10–173 weeks) and (1–646 weeks), respectively]. The median overall survival was 41 weeks vs. 60 weeks for patients receiving ATRA or not [(p=0.74), ranges, (3–452 weeks) and (0–735 weeks), respectively]. Conclusion: There was no significant difference in CR rate, EFS, or OS in this higher risk population of patients with NPM1 mutated diploid AML or high-risk MDS receiving ATRA as an adjunct to chemotherapy. Disclosures: Kantarjian: Genzyme: Research Funding. Ravandi:genzyme: Research Funding.

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Quantification of Fosfomycin in Combination with Nine Antibiotics in Human Plasma and Cation-Adjusted Mueller-Hinton II Broth via LCMS

Antibiotics ◽

10.3390/antibiotics11010054 ◽

2022 ◽

Vol 11 (1) ◽

pp. 54

Author(s):

Kelvin Kau-Kiat Goh ◽

Wilson Ghim-Hon Toh ◽

Daryl Kim-Hor Hee ◽

Edwin Zhi-Wei Ting ◽

Nathalie Grace Sy Chua ◽

...

Keyword(s):

Combination Therapy ◽

Human Plasma ◽

Limit Of Detection ◽

Synergistic Activity ◽

Limit Of Quantification ◽

Mueller Hinton ◽

Antibiotic Drug ◽

Dosing Regimens ◽

Antibiotic Combination

Fosfomycin-based combination therapy has emerged as an attractive option in our armamentarium due to its synergistic activity against carbapenem-resistant Gram-negative bacteria (CRGNB). The ability to simultaneously measure fosfomycin and other antibiotic drug levels will support in vitro and clinical investigations to develop rational antibiotic combination dosing regimens against CRGNB infections. We developed an analytical assay to measure fosfomycin with nine important antibiotics in human plasma and cation-adjusted Mueller–Hinton II broth (CAMHB). We employed a liquid-chromatography tandem mass spectrometry method and validated the method based on accuracy, precision, matrix effect, limit-of-detection, limit-of-quantification, specificity, carryover, and short-term and long-term stability on U.S. Food & Drug Administration (FDA) guidelines. Assay feasibility was assessed in a pilot clinical study in four patients on antibiotic combination therapy. Simultaneous quantification of fosfomycin, levofloxacin, meropenem, doripenem, aztreonam, piperacillin/tazobactam, ceftolozane/tazobactam, ceftazidime/avibactam, cefepime, and tigecycline in plasma and CAMHB were achieved within 4.5 min. Precision, accuracy, specificity, and carryover were within FDA guidelines. Fosfomycin combined with any of the nine antibiotics were stable in plasma and CAMHB up to 4 weeks at −80 °C. The assay identified and quantified the respective antibiotics administered in the four subjects. Our assay can be a valuable tool for in vitro and clinical applications.

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Structurally Simple Phenanthridine Analogues Based on Nitidine and Their Antitumor Activities

Molecules ◽

10.3390/molecules24030437 ◽

2019 ◽

Vol 24 (3) ◽

pp. 437

Author(s):

Shu-Qin Qin ◽

Lian-Chun Li ◽

Jing-Ru Song ◽

Hai-Yun Li ◽

Dian-Peng Li

Keyword(s):

Anticancer Agents ◽

Human Cancer ◽

A549 Cells ◽

Anticancer Activities ◽

Human Cancer Cell Lines ◽

Ic50 Value ◽

Ic50 Values ◽

Diphenyl Tetrazolium Bromide ◽

Conjugated Compounds

A series of novel structurally simple analogues based on nitidine was designed and synthesized in search of potent anticancer agents. The antitumor activity against human cancer cell lines (HepG2, A549, NCI-H460, and CNE1) was performed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay in vitro. The results showed that some of them had good anticancer activities, especially derivatives with a [(dimethylamino)ethyl]amino side chain in the C-6 position. Planar conjugated compounds 15a, 15b, and 15c, with IC50 values of 1.20 μM, 1.87 μM, and 1.19 μM against CNE1 cells, respectively, were more active than nitidine chloride. Compound 15b and compound 15c with IC50 values of 1.19 μM and 1.37 μM against HepG2 cells and A549 cells demonstrated superior activities to nitidine. Besides, compound 5e which had a phenanthridinone core displayed extraordinary cytotoxicity against all test cells, particularly against CNE1 cells with the IC50 value of 1.13 μM.

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Modulation of spontaneous B-cell differentiation in macroglobulinemia by retinoic acid

Blood ◽

10.1182/blood.v83.8.2206.2206 ◽

1994 ◽

Vol 83 (8) ◽

pp. 2206-2210 ◽

Cited By ~ 6

Author(s):

Y Levy ◽

S Labaume ◽

MC Gendron ◽

JC Brouet

Keyword(s):

Retinoic Acid ◽

Cell Differentiation ◽

B Cells ◽

B Cell ◽

Plasma Cells ◽

B Cell Differentiation ◽

Trans Retinoic Acid ◽

All Trans Retinoic Acid ◽

Culture Supernatants

Abstract We previously showed that clonal blood B cells from patients with macroglobulinemia spontaneously differentiate in vitro to plasma cells. This process is dependent on an interleukin (IL)-6 autocrine pathway. We investigate here whether all-trans-retinoic acid (RA) interferes with B-cell differentiation either in patients with IgM gammapathy of undetermined significance (MGUS) or Waldenstrom's macroglobulinemia (WM). RA at a concentration of 10(-5) to 10(-8) mol/L inhibited by 50% to 80% the in vitro differentiation of purified B cells from four of five patients with MGUS and from one of five patients with WM as assessed by the IgM content of day 7 culture supernatants. We next determined whether this effect could be related to an inhibition of IL- 6 secretion by cultured B cells and/or a downregulation of the IL-6 receptor (IL-6R), which was constitutively expressed on patients' blood B cells. A 50% to 100% (mean, 80%) inhibition of IL-6 production was found in seven of 10 patients (five with MGUS and two with WM). The IL- 6R was no more detectable on cells from patients with MGUS after 2 days of treatment with RA and slightly downregulated in patients with WM. It was of interest that B cells susceptible to the action of RA belonged mostly to patients with IgM MGUS, which reinforces our previous data showing distinct requirements for IL-6-dependent differentiation of blood B cells from patients with VM or IgM MGUS.

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Momilactones A, B, and Tricin in Rice Grain and By-Products are Potential Skin Aging Inhibitors

Foods ◽

10.3390/foods8120602 ◽

2019 ◽

Vol 8 (12) ◽

pp. 602 ◽

Cited By ~ 3

Author(s):

Nguyen Van Quan ◽

Dam Duy Thien ◽

Tran Dang Khanh ◽

Hoang-Dung Tran ◽

Tran Dang Xuan

Keyword(s):

Liquid Chromatography ◽

High Performance ◽

Biological Activities ◽

Ultra Performance Liquid Chromatography ◽

Skin Aging ◽

Ionization Mass ◽

Rice Grain ◽

Synergistic Activity ◽

Ic50 Values

We previously reported the inhibitory potentials of momilactones A (MA) and B (MB) against key enzymes related to type 2 diabetes and obesity. In this study, antioxidant and anti-skin-aging activities of MA and MB were investigated and compared with tricin, a well-known antioxidant and antiaging flavonoid in rice. MA, MB, and tricin were purified from rice husk by column chromatography and their biological activities were subsequently assayed by in vitro trials. The contents of MA, MB, and tricin of different commercial rice cultivars in Japan were quantified and confirmed by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) and high-performance liquid chromatography (HPLC) analyses. The antioxidant assays revealed a synergistic activity of the mixture MA and MB (MAB, 1:1, v/v). In addition, in 2,2’-azino-bis (ABTS) assay, IC50 values of MAB (0.3 mg/mL) and tricin (0.3 mg/mL) was 4-fold and 9-fold greater than that of individual MB (1.3 mg/mL) or MA (2.8 mg/mL), respectively. The in vitro enzymatic assays on pancreatic elastase and tyrosinase indicated that MA and MB were potential to relief skin wrinkles and freckles. In detail, MA exerted higher inhibition on both enzymatic activities (30.9 and 37.6% for elastase and tyrosinase inhibition, respectively) than MB (18.5 and 12.6%) and MAB (32.0 and 19.7%) at a concentration of 2.0 mg/mL. Notably, MA and the mixture MAB exhibited stronger inhibitions on elastase and tyrosinase in comparison with tricin and vanillin. MA, MB, and tricin in rice are potential to develop cosmetics as well as supplements for skin aging treatments.

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α-Glucosidase Inhibition and Molecular Docking Studies of Natural Brominated Metabolites from Marine Macro Brown Alga Dictyopteris hoytii

Marine Drugs ◽

10.3390/md17120666 ◽

2019 ◽

Vol 17 (12) ◽

pp. 666 ◽

Cited By ~ 8

Author(s):

Najeeb Ur Rehman ◽

Kashif Rafiq ◽

Ajmal Khan ◽

Sobia Ahsan Halim ◽

Liaqat Ali ◽

...

Keyword(s):

Molecular Docking ◽

Brown Alga ◽

Docking Studies ◽

Spectroscopic Techniques ◽

Molecular Docking Studies ◽

Ethyl Methyl ◽

2D Noesy ◽

Ic50 Value ◽

Ic50 Values

Bioassay guided isolation of the methanolic extract of marine macro brown alga Dictyopteris hoytii afforded one new metabolite (ethyl methyl 2-bromobenzene 1,4-dioate, 1), one new natural metabolite (diethyl-2-bromobenzene 1,4-dioate, 2) along with six known metabolites (3–8) reported for the first time from this source. The structure elucidation of all these compounds was achieved by extensive spectroscopic techniques including 1D (1H and 13C) and 2D (NOESY, COSY, HMBC and HSQC) NMR and mass spectrometry and comparison of the spectral data of known compounds with those reported in literature. The in vitro α-glucosidase inhibition studies confirmed compound 7 to be the most active against α-glucosidase enzyme with IC50 value of 30.5 ± 0.41 μM. Compounds 2 and 3 demonstrated good inhibition with IC50 values of 234.2 ± 4.18 and 289.4 ± 4.91 μM, respectively, while compounds 1, 5, and 6 showed moderate to low inhibition. Furthermore, the molecular docking studies of the active compounds were performed to examine their mode of inhibition in the binding site of the α-glucosidase enzyme.

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In VitroCombination of Isavuconazole with Micafungin or Amphotericin B Deoxycholate against Medically Important Molds

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.03261-14 ◽

2014 ◽

Vol 58 (11) ◽

pp. 6934-6937 ◽

Cited By ~ 25

Author(s):

Aspasia Katragkou ◽

Matthew McCarthy ◽

Joseph Meletiadis ◽

Vidmantas Petraitis ◽

Patriss W. Moradi ◽

...

Keyword(s):

Combination Therapy ◽

Invasive Aspergillosis ◽

Amphotericin B ◽

Aspergillus Flavus ◽

Aspergillus Terreus ◽

Synergistic Activity ◽

Content Type ◽

Amphotericin B Deoxycholate ◽

Combination Studies

ABSTRACTWhether isavuconazole, an extended-spectrum triazole, possesses synergistic activity in combination therapy with echinocandins or amphotericin B for the treatment of invasive molds infections has not been studied. Ourin vitrocombination studies showed that isavuconazole and micafungin are synergistically active againstAspergillus fumigatus,Aspergillus flavus,Aspergillus terreus, andCunninghamella bertholletiae. These results suggest that isavuconazole, in combination with micafungin, may have a role in the treatment of invasive aspergillosis and warrants further investigation.

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Inhibition of Androgen Synthesis in Human Testicular and Prostatic Microsomes and in Male Rats by Novel Steroidal Compounds*

Endocrinology ◽

10.1210/endo.140.6.6832 ◽

1999 ◽

Vol 140 (6) ◽

pp. 2891-2897 ◽

Cited By ~ 8

Author(s):

Ivo P. Nnane ◽

Katsuya Kato ◽

Yang Liu ◽

Brian J. Long ◽

Qing Lu ◽

...

Keyword(s):

Male Rats ◽

Normal Male ◽

Rat Tissues ◽

The Novel ◽

Androgen Synthesis ◽

Potent Inhibition ◽

Ic50 Value ◽

Ic50 Values ◽

Wet Weight

Abstract The C17,20-lyase and 5α-reductase are key enzymes in the biosynthesis of androgens. The effects of novel steroidal compounds were evaluated as inhibitors against both human C17,20-lyase and 5α-reductase in vitro. The concentrations of testosterone (T) and dihydrotestosterone (DHT) in the prostate, testis and serum and changes in the tissue weights were also determined in rats treated with the novel inhibitors. L-12 and L-26 showed potent inhibition of human testicular C17,20-lyase with IC50 values of 50 and 25 nm, respectively. L-12, L-38, and I-47 showed moderate inhibition of human testicular C17,20-lyase with IC50 values of 75, 108, and 70 nm, respectively similar to ketoconazole (78 nm). Interestingly, L-6, L-26, and L-38 also showed some inhibitory activity against 5α-reductase with IC50 values of 75, 125, and 377 nm, respectively. Finasteride, an inhibitor of 5α-reductase had an IC50 value of 33 nm. However, ketoconazole did not inhibit 5α-reductase nor did finasteride inhibit C17,20-lyase. Treatment of normal male rats with several of these novel inhibitors (50 mg/kg·day, sc, for 14 consecutive days) caused about 45–91% decrease in serum, testicular and prostatic T concentration. Similarly, serum and prostatic DHT concentration were significantly decreased in rats treated with these novel compounds by 50–90% compared with controls. Surgical castration caused almost complete elimination of circulating T and DHT concentration in rat tissues. L-6 and L-12 were the most effective and reduced the wet weight of the prostate by 50%. Although future improvements in their bioavailability are necessary, these novel steroidal compounds show promise as potential agents for reducing T and DHT levels in patients with androgen dependent diseases.

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Sintesis dan Evaluasi Antimalaria In Vitro Turunan Kinin Terhadap Plasmodium falciparum

Jurnal Kefarmasian Indonesia ◽

10.22435/jki.v11i2.3923 ◽

2021 ◽

Vol 11 (2) ◽

pp. 109-120

Author(s):

Salahuddin Salahuddin ◽

Rahmana Emran K ◽

Muhammad Hanafi ◽

Andini Sundowo ◽

Puspa Dewi NL ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Antimalarial Drug ◽

Malaria Treatment ◽

Reaction Products ◽

Synthetic Product ◽

13C Nuclear Magnetic Resonance ◽

Ic50 Value ◽

Ic50 Values

Nowadays kinin is the most effective antimalarial drug and its used as an alternative in malaria treatment. However, toxicity of quinine restrict its use as an antimalarial drug. Lipophilicity and long half-life (t½) of quinine that reach 10-20 hours are responsible for its toxicity. The aim of this research is to obtain more polar quinine derivatives by means of hydrogen peroxide reactions to reduce the toxicity. The reactions using hydrogen peroxyde is performed analogously to the procedures reported in the literature. Extract of pure anhydrous kinin is purified in coloumn chromatography followed by structure elucidation. Synthetic product is tested in vitro against Plasmodium falciparum. The characterization of reaction products is performed with proton (1H) and carbon 13 (13C) nuclear magnetic resonance (NMR) spectroscopy. It showed that the reaction using reagents led to epoxidation of vinyl substituents of chinuclidine ring with 61,08% yields. Antimalarial test against Plasmodium falciparum obtained 1.250-2.500 μg/mL of IC50 value. The IC50 values indicated that the synthesis products were not potential for malaria treatment.

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Caspase-8, RIPK1, and RIPK3 Coordinately Regulate Retinoic Acid-Induced Cell Differentiation and Necroptosis

10.1101/156901 ◽

2017 ◽

Author(s):

Masataka Someda ◽

Shunsuke Kuroki ◽

Makoto Tachibana ◽

Shin Yonehara

Keyword(s):

Retinoic Acid ◽

Cell Differentiation ◽

Es Cells ◽

Death Receptor ◽

Binding Activity ◽

Embryoid Bodies ◽

Caspase 8 ◽

Retinoic Acid Response Element

AbstractCaspase-8, which is essential for death receptor-mediated apoptosis, inhibits necroptosis by suppressing the function of RIPK1 and RIPK3 to activate MLKL. We show that knockdown of caspase-8 expression in embryoid bodies derived from ES cells markedly enhances retinoic acid (RA)-induced cell differentiation and necroptosis, both of which are dependent on Ripkl and Ripk3. RA treatment obviously enhanced the expression of RA-specific genes having a retinoic acid response element (RARE) to induce cell differentiation, and induced marked expression of RIPK1, RIPK3 and MLKL to stimulate necroptosis. Caspase-8 knockdown induced RA receptor (RAR) to form a complex with RIPK1 and RIPK3 in the nucleus, and RAR interacting with RIPK1 and RIPK3 showed much stronger binding activity to RARE than RAR without RIPK1 or RIPK3. In Caspase-8-deficient mouse embryos, expression of RA-specific genes was obviously enhanced. Thus, caspase-8, RIPK1, and RIPK3 regulate RA-induced cell differentiation and necroptosis both in vitro and in vivo.

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