-
Molecules (Basel, Switzerland) Mar 2021Osteomyelitis and orthopedic infections are major clinical problems, limited by a lack of antibiotics specialized for such applications. In this paper, we describe the...
Osteomyelitis and orthopedic infections are major clinical problems, limited by a lack of antibiotics specialized for such applications. In this paper, we describe the design and synthesis of a novel bone-binding antibiotic (BBA-1) and its subsequent structural and functional characterization. The synthesis of BBA-1 was the result of a two-step chemical conjugation of cationic selective antimicrobial-90 (CSA-90) and the bisphosphonate alendronate (ALN) via a heterobifunctional linker. This was analytically confirmed by HPLC, FT-IR, MS and NMR spectroscopy. BBA-1 showed rapid binding and high affinity to bone mineral in an in vitro hydroxyapatite binding assay. Kirby-Baur assays confirmed that BBA-1 shows a potent antibacterial activity against and methicillin-resistant comparable to CSA-90. Differentiation of cultured osteoblasts in media supplemented with BBA-1 led to increased alkaline phosphatase expression, which is consistent with the pro-osteogenic activity of CSA-90. Bisphosphonates, such as ALN, are inhibitors of protein prenylation, however, the amine conjugation of ALN to CSA-90 disrupted this activity in an in vitro protein prenylation assay. Overall, these findings support the antimicrobial, bone-binding, and pro-osteogenic activities of BBA-1. The compound and related agents have the potential to ensure lasting activity against osteomyelitis after systemic delivery.
Topics: 3T3 Cells; Alendronate; Animals; Anti-Bacterial Agents; Anti-Infective Agents; Bone and Bones; Calcification, Physiologic; Cell Differentiation; Cells, Cultured; Diphosphonates; Methicillin-Resistant Staphylococcus aureus; Mice; Osteoblasts; Osteogenesis; Osteomyelitis; Pregnanes; Propylamines; Spectroscopy, Fourier Transform Infrared; Staphylococcal Infections; Staphylococcus aureus
PubMed: 33799713
DOI: 10.3390/molecules26061541 -
The Journal of Pharmacology and... Apr 2016Synaptic GABAA receptors are primary mediators of rapid inhibition in the brain and play a key role in the pathophysiology of epilepsy and other neurologic disorders....
Synaptic GABAA receptors are primary mediators of rapid inhibition in the brain and play a key role in the pathophysiology of epilepsy and other neurologic disorders. The δ-subunit GABAA receptors are expressed extrasynaptically in the dentate gyrus and contribute to tonic inhibition, promoting network shunting as well as reducing seizure susceptibility. However, the neurosteroid structure-function relationship at δGABA(A) receptors within the native hippocampus neurons remains unclear. Here we report a structure-activity relationship for neurosteroid modulation of extrasynaptic GABAA receptor-mediated tonic inhibition in the murine dentate gyrus granule cells. We recorded neurosteroid allosteric potentiation of GABA as well as direct activation of tonic currents using a wide array of natural and synthetic neurosteroids. Our results shows that, for all neurosteroids, the C3α-OH group remains obligatory for extrasynaptic receptor functional activity, as C3β-OH epimers were inactive in activating tonic currents. Allopregnanolone and related pregnane analogs exhibited the highest potency and maximal efficacy in promoting tonic currents. Alterations at the C17 or C20 region of the neurosteroid molecule drastically altered the transduction kinetics of tonic current activation. The androstane analogs had the weakest modulatory response among the analogs tested. Neurosteroid potentiation of tonic currents was completely (approximately 95%) diminished in granule cells from δ-knockout mice, suggesting that δ-subunit receptors are essential for neurosteroid activity. The neurosteroid sensitivity of δGABA(A) receptors was confirmed at the systems level using a 6-Hz seizure test. A consensus neurosteroid pharmacophore model at extrasynaptic δGABA(A) receptors is proposed based on a structure-activity relationship for activation of tonic current and seizure protection.
Topics: Animals; Anticonvulsants; Dentate Gyrus; GABA Agonists; GABA Modulators; In Vitro Techniques; Interneurons; Mice; Mice, Inbred C57BL; Mice, Knockout; Midazolam; Neurons; Neurotransmitter Agents; Pregnanes; Pregnanolone; Receptors, GABA-A; Seizures; Structure-Activity Relationship
PubMed: 26857959
DOI: 10.1124/jpet.115.229302 -
International Journal of Molecular... Jun 2022Progesterone biotransformation is worth studying because of the high industrial value of its derivatives. This study investigated the catalytic ability of the...
Progesterone biotransformation is worth studying because of the high industrial value of its derivatives. This study investigated the catalytic ability of the entomopathogenic filamentous fungus strain KCh KW1.1 to transform progesterone derivatives: 11α-hydroxyprogesterone, 17α-hydroxyprogesterone, 16α,17α-epoxyprogesterone and pregnenolone. In the culture of KCh KW1.1, 11α-hydroxyprogesterone was effectively transformed into only one product: 6β,11α-dihydroxyprogesterone. Transformation of 17α-hydroxyprogesterone gave three hydroxy derivatives: 6β,17α-dihydroxyprogesterone, 12β,17α-dihydroxyprogesterone and 6β,12β,17α-trihydroxyprogesterone. Two products: 6β-hydroxy-16α,17α-epoxyprogesterone and 6β,11α-dihydroxy-16α,17α-epoxyprogesterone, were obtained from the 16α,17α-epoxyprogesterone transformation. We isolated two compounds from the biotransformation medium with pregnenolone: 11α-hydroxy-7-oxopregnenolone and 5α,6α-epoxy-3β,11α-dihydroxypregnan-7,20-dione. In this study, we observed only mono- and dihydroxy derivatives of the tested substrates, and the number of obtained products for each biotransformation did not exceed three.
Topics: Algestone; Biotransformation; Cordyceps; Hydroxylation; Hydroxyprogesterones; Pregnenolone; Progesterone
PubMed: 35806021
DOI: 10.3390/ijms23137015 -
The American Journal of Sports Medicine Jul 2023The chondrotoxic effects of methylprednisolone acetate (MP) and triamcinolone acetonide (TA) have been well described. However, the mechanical effects of these commonly...
BACKGROUND
The chondrotoxic effects of methylprednisolone acetate (MP) and triamcinolone acetonide (TA) have been well described. However, the mechanical effects of these commonly used steroids on native cartilage are largely unknown.
PURPOSE
To investigate the in vitro effects of a single 1-hour MP or TA exposure on the viability, mechanics, and biochemical content of native articular cartilage explants.
STUDY DESIGN
Controlled laboratory study.
METHODS
Articular cartilage explants (n = 6 per group) were harvested from the femoral condyles of bovine stifles. Explants were exposed to chondrogenic medium containing a clinical dose of MP or TA for 1 hour, followed by fresh medium wash and exchange. Explants in the control group underwent the same treatment with chondrogenic medium alone. At 24 hours after treatment, samples were assessed for viability (live/dead), mechanical properties (creep indentation and Instron tensile testing), biochemical (collagen and glycosaminoglycan) content, and pyridinoline crosslinking via mass spectrometry.
RESULTS
Mean cell viability was significantly decreased in native explants exposed to MP (35.5%) compared with the control (49.8%; < .001) and TA (45.7%; = .01) specimens. Significant decreases were seen in the mechanical properties of steroid-treated native explants when compared with controls, with decreases in aggregate modulus (646.3 vs 312.8 kPa [MP] and 257.0 kPa [TA]; < .001), shear modulus (370.1 vs 191.2 kPa [MP] and 157.4 kPa [TA]; < .001), and ultimate tensile strength (9.650 vs 5.648 MPa [MP; = .021] and 6.065 MPa [TA; = .0403]). No significant differences in collagen and glycosaminoglycan content were found in the steroid-treated groups. Pyridinoline crosslinking was significantly decreased in explants exposed to TA compared with controls ( = .027).
CONCLUSION
Exposure of MP to articular cartilage explants was chondrotoxic, and exposure of articular cartilage explants to MP or TA resulted in significant decreases in mechanical properties of articular cartilage explants compared with controls. Clinicians should be judicious regarding use of intra-articular steroids, particularly in patients with intact healthy articular cartilage.
Topics: Humans; Animals; Cattle; Methylprednisolone; Triamcinolone; Cartilage, Articular; Triamcinolone Acetonide; Methylprednisolone Acetate; Glycosaminoglycans
PubMed: 37183987
DOI: 10.1177/03635465231162644 -
Cells Oct 2022Triazole fungicides such as propiconazole (Pi) or tebuconazole (Te) show hepatotoxicity in vivo, e.g., hypertrophy and vacuolization of liver cells following interaction...
New Approach Methods for Hazard Identification: A Case Study with Azole Fungicides Affecting Molecular Targets Associated with the Adverse Outcome Pathway for Cholestasis.
Triazole fungicides such as propiconazole (Pi) or tebuconazole (Te) show hepatotoxicity in vivo, e.g., hypertrophy and vacuolization of liver cells following interaction with nuclear receptors such as PXR (pregnane-X-receptor) and CAR (constitutive androstane receptor). Accordingly, azoles affect gene expression associated with these adverse outcomes in vivo but also in human liver cells in vitro. Additionally, genes indicative of liver cholestasis are affected in vivo and in vitro. We therefore analyzed the capability of Pi and Te to cause cholestasis in an adverse outcome pathway (AOP)-driven approach in hepatic cells of human origin in vitro, considering also previous in vivo studies. Bile salt export pump (BSEP) activity assays confirmed that both azoles are weak inhibitors of BSEP. They alternate the expression of various cholestasis-associated target genes and proteins as well as the mitochondrial membrane function. Published in vivo data, however, demonstrate that neither Pi nor Te cause cholestasis in rodent bioassays. This discrepancy can be explained by the in vivo concentrations of both azoles being well below their EC50 for BSEP inhibition. From a regulatory perspective, this illustrates that toxicogenomics and human in vitro models are valuable tools to detect the potential of a substance to cause a specific type of toxicity. To come to a sound regulatory conclusion on the in vivo relevance of such a finding, results will have to be considered in a broader context also including toxicokinetics in a weight-of-evidence approach.
Topics: Humans; ATP Binding Cassette Transporter, Subfamily B, Member 11; Fungicides, Industrial; Adverse Outcome Pathways; Azoles; Cholestasis; Receptors, Cytoplasmic and Nuclear; Triazoles; Pregnanes
PubMed: 36291160
DOI: 10.3390/cells11203293 -
Journal of Affective Disorders Apr 2021Zuranolone (SAGE-217) is a novel, investigational positive allosteric modulator of GABA receptors being investigated in major depressive disorder (MDD). This analysis of... (Meta-Analysis)
Meta-Analysis
BACKGROUND
Zuranolone (SAGE-217) is a novel, investigational positive allosteric modulator of GABA receptors being investigated in major depressive disorder (MDD). This analysis of phase 2 data quantified the benefit and risk of zuranolone (30mg) versus placebo and antidepressants in terms of number needed to treat (NNT) and number needed to harm (NNH).
METHODS
Rates of response, remission, and all-cause discontinuation for zuranolone and 11 antidepressant comparators were obtained from the zuranolone phase 2 clinical study (N=89) and a published network meta-analysis, respectively. An indirect treatment comparison was conducted using the Bucher method to compare zuranolone to standard-of-care.
RESULTS
Zuranolone demonstrated greater benefit compared to placebo on Day 3 (NNT range for response=4-5, NNT for remission=10) and at Day 15 (NNT=3 for response and remission). Compared to SSRIs and SNRIs, zuranolone at Day 15 showed improved treatment response (NNT=4 [95% CI = 3; 16] and 5 [95% CI = 3; 25], respectively) and remission (NNT=4 [95% CI = 2; 13] and 4 [95% CI = 2; 18], respectively). This was accompanied by a reduction in all-cause discontinuation, with negative NNH values (-57 and -28), respectively.
LIMITATIONS
Variations in study design across the included trials may limit the generalizability of results.
CONCLUSIONS
With a small positive NNT as early as Day 3 indicating robust benefit and a negative NNH indicating reduced harm, this analysis based on a phase 2 study suggests that patients with MDD may benefit from the benefit-to-risk profile of zuranolone.
Topics: Antidepressive Agents; Clinical Trials, Phase II as Topic; Depressive Disorder, Major; Humans; Pregnanes; Pyrazoles
PubMed: 33640861
DOI: 10.1016/j.jad.2021.02.027 -
Biomedicine & Pharmacotherapy =... Jul 2020As ongoing investigation of Huernia saudi-arabica D.V.Field (Asclepiadaceae), a new steroidal pregnane glycoside (Huernioside A) was isolated from dichloromethane...
As ongoing investigation of Huernia saudi-arabica D.V.Field (Asclepiadaceae), a new steroidal pregnane glycoside (Huernioside A) was isolated from dichloromethane fraction (DCM); it was identified as 3β, 11, 14β, 20(R)-tetrahydroxy-pregna-5,9(11)-diene-3-O-β-D-thevetopyranosyl-(1-4)-β-D-cymaropyranoside(HCP) through analysis of 1D, 2D NMR besides ESI-MS data. The alcoholic extract of the aerial part (ALE), DCM and HCP showed inhibitory potential against pancreatic lipase compared to orilstat. Among the tested samples, the ALE and HCP exhibited a promising pancreatic lipase inhibitory commotion through IC values of 0.61 ± 0.15, 1.23 ± 0.07 mg/ml (equivalent to 88.8 μM), respectively. HCP was prevailed to have a mixed mode of inhibition as exposed by enzyme kinetic studies. Hydrophobic interactions were the major forces involved in ligand enzyme interactions. In contrast, moderate α-glucosidase inhibitory activities were evidenced for ALE and HCP (% inhibition: 24.8 ± 1.8 and 26.6 ± 2.5, respectively) compared to acarbose. This investigation is the first to report on the possible in vitro anti-obesity and anti-diabetic impact of H. saudi-arabica.
Topics: Apocynaceae; Glycoside Hydrolase Inhibitors; Glycosides; Lipase; Plant Extracts; Pregnanes
PubMed: 32339923
DOI: 10.1016/j.biopha.2020.110143 -
Molecules (Basel, Switzerland) Oct 2022Metastasis is an important cause of cancer-related death. Previous studies in our laboratory found that pregnane alkaloids from had antimetastatic activity against...
Metastasis is an important cause of cancer-related death. Previous studies in our laboratory found that pregnane alkaloids from had antimetastatic activity against breast cancer cells. In the current study, we demonstrated that treatment with one of the alkaloid derivatives, ()-3β-ethylamino-pregn-17(20)-en (), led to the downregulation of the HIF-1α/VEGF/VEGFR2 pathway, suppressed the phosphorylation of downstream molecules Akt, mTOR, FAK, and inhibited breast cancer metastasis and angiogenesis both in vitro and in vivo. Furthermore, the antimetastasis and antiangiogenesis effects of treatment (40 mg/kg) were more effective than that of Sorafenib (50 mg/kg). Surface plasmon resonance (SPR) analysis was performed and the result suggested that HSP90α was a direct target of . Taken together, our results suggested that compound might represent a candidate antitumor agent for metastatic breast cancer.
Topics: Humans; Angiogenesis Inhibitors; Proto-Oncogene Proteins c-akt; Vascular Endothelial Growth Factor A; Triple Negative Breast Neoplasms; Sorafenib; Cell Line, Tumor; Neovascularization, Pathologic; Antineoplastic Agents; TOR Serine-Threonine Kinases; Alkaloids; Pregnanes; Hypoxia-Inducible Factor 1, alpha Subunit
PubMed: 36296726
DOI: 10.3390/molecules27207132 -
Proceedings of the Royal Society of... Aug 1965
Topics: Asthma; Betamethasone; Drug Therapy; Humans; Prednisone; Research; Steroids; Triamcinolone
PubMed: 14341858
DOI: No ID Found -
Biological & Pharmaceutical Bulletin 2022Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is a potential target for inflammatory-breast cancer treatment as it participates in its...
Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is a potential target for inflammatory-breast cancer treatment as it participates in its pathogenesis, such as tumor initiation, progression, survival, metastasis, and recurrence. In this study, we aimed to discover a novel anti-cancer treatment from natural products by targeting NF-κB activity. Using the 4T1-NFκB-luciferase reporter cell line, we tested three pregnane glycosides extracted from the herb Caralluma tuberculata and discovered that Russelioside A markedly suppressed NF-κB activity in breast cancer. Russelioside A inhibited NF-κB (p65) transcriptional activity and its phosphorylation. Following NF-κB inhibition, Russelioside A exerted anti-proliferative and anti-metastatic effects in breast cancer cells in vitro. Moreover, it inhibited the NF-κB constitutive expression of downstream pathways, such as VEGF-b, MMP-9, and IL-6 in 4T1 cells. In addition, it reduced the metastatic capacity in a 4T1 breast cancer model in vivo. Collectively, our conclusions reveal that Russelioside A is an attractive natural compound for treating triple-negative breast cancer growth and metastasis through regulating NF-κB activation.
Topics: Apocynaceae; Biological Products; Breast Neoplasms; Cell Line, Tumor; Female; Glycosides; Humans; Interleukin-6; Matrix Metalloproteinase 9; NF-kappa B; Pregnanes; Triple Negative Breast Neoplasms; Vascular Endothelial Growth Factor B
PubMed: 36184517
DOI: 10.1248/bpb.b22-00508