incyclinide  (MetaStat Inc)

Journal: bioRxiv

Article Title: Atypical tetracyclines promote longevity and ferroptotic neuroprotection via translation attenuation

doi: 10.64898/2026.01.09.698733

Figure Lengend Snippet: (A) Shown is the general structure of the tetracyclines with the keto-enol system (pink) at C11 & C12 that is responsible for the chelation of Zn 2+ and other divalent ions. Structural modifications in 12-aminominocycline and R464 disrupt the chelation center (blue & orange circles). (B) Colorimetric assay measuring the % remaining Zn 2+ as a function of tetracycline concentration, indicative of the chelation effect of each tetracycline. 12-aminominocycline does not chelate ions at concentrations up to 1800 µM, whereas 300 µM is required for R464. Other tetracyclines tested (grey) include: 4-epiminocycline, minocycline, Col-3, Doxycycline, Tigecycline, & Evaracycline. EDTA was used as a positive control. (C) Bar graph shows the % remaining recombinant MMP9 activity after tetracycline treatment (100 µM). The assay measures proteolytic cleavage of a fluorogenic substrate, released upon cleavage. NNGH is a broad-spectrum inhibitor of matrix metalloproteinases and was used as a positive control. Significance was determined by one-way ANOVA with Dunnett’s multiple comparisons, where *** = p < 0.001 and **** = p < 0.0001. Error bars indicate mean ± SD from three independent trials. (D) Dose response curve of four neuroprotective tetracyclines. Tetracyclines with IC 50 greater than 300 µM are indicated as “not determined” (n.d.).

Article Snippet: Tetracyclines: Minocycline (MP Biomedicals, Cat#155718), 4-epiminocycline (Toronto Research Chemicals, Cat#TRC-E588540), Doxycycline (Sigma, Cat#D9891), Col-3 (Incyclinide, MedChemExpress, Cat#HY-13648), R464449 (Sigma-Aldrich, Cat#R464449), 12-aminominocycline (Toronto Research Chemicals, Cat#TRC-A618285), Tigecycline (LKT Laboratories, Cat#T3324).

Techniques: Colorimetric Assay, Concentration Assay, Positive Control, Recombinant, Activity Assay

Journal: bioRxiv

Article Title: Combining Top-Down and Bottom-Up: An Open Microfluidic Microtumor Model for Investigating Tumor Cell-ECM Interaction and Anti-Metastasis

doi: 10.1101/2024.03.19.585776

Figure Lengend Snippet: Anti-metastasis drug testing. (a) A schematic illustrates the workflow of in vivo mice experiments (see Experimental Section). (b) Primary tumor mass shows no significant differences between the PBS [i.e., control (-)] and incyclinide-treated mice groups. (c) The number of metastatic colonies from the incyclinide-treated group is significantly lower than that of the PBS group. (d) A schematic demonstrating the workflow of UOMS microtumor model test with incyclinide treatment (see Experimental Section). (e) The representative composite microscopic images from the UOMS microtumor model (2-2- L 0.5- W 0.1) test with and without incyclinide treatment (Movie S3). (f), (g) ECM contraction and cell count analyses of (e). On UOMS microtumor model, tumor cells treated with incyclinide displayed reduced migration and ECM contraction. Error bars are mean ± S.D. from ≥3 replicates. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001. “ns” represents “not significant”.

Article Snippet: 40 mg of incyclinide (TA9H94533373, Sigma-Aldrich) was dissolved in 1 mL dimethyl sulfoxide (DMSO) to reach 40 μg/μL.

Techniques: In Vivo, Cell Counting, Migration