Discovery of HyT-Based Degraders of CDK9-Cyclin T1 Complex.

Direct modulation of the non-kinase functions of cyclin and CDK-cyclin complexes poses challenges. We utilize hydrophobic tag (HyT) based small-molecule degraders induced degradation of cyclin T1 and its corresponding kinase partner CDK9. LL-CDK9-12 demonstrated the most potent and selective degradation ability, with DC50 values of 0.362 µM against CDK9 and 0.680 µM against cyclin T1. In prostate cancer cells, LL-CDK9-12 showed enhanced anti-proliferative activity than its parental molecule SNS032 and LL-K9-3, the previous reported CDK9-cyclin T1 degrader. Moreover, LL-CDK9-12 suppressed the downstream signaling of CDK9 and AR efficiently. Altogether, LL-CDK9-12 was an effective dual degrader of CDK9-cyclin T1 and helped study the unknown function of CDK9-cyclin T1. These results suggest that HyT-based degraders could be used as a strategy to induce the degradation of protein complexes, providing insights for the design of protein complexes' degraders.

Discovery of LL-K8-22: A Selective, Durable, and Small-Molecule Degrader of the CDK8-Cyclin C Complex.

The CDK8-cyclin C complex is an important anti-tumor target, but unlike CDK8, cyclin C remains undruggable. Modulators regulating cyclin C activity directly are still under development. Here, a series of hydrophobic tagging-based degraders of the CDK8-cyclin C complex were designed, synthesized, and evaluated to identify the first dual degrader, LL-K8-22, which induced selective and synchronous degradation of CDK8 and cyclin C. Proteomic and immunoblot studies exhibited that LL-K8-22 significantly degraded CDK8 without reducing CDK19 and did not degrade other cyclin proteins except cyclin C. Moreover, LL-K8-22 showed enhanced anti-proliferative effects over its parental molecule, BI-1347, with potency increased by 5-fold in MDA-MB-468 cells. LL-K8-22 exhibited more pronounced effects on CDK8-cyclin C downstream signaling than BI-1347, suppressing STAT1 phosphorylation more persistently. RNA-sequencing analysis revealed that LL-K8-22 inhibited E2F- and MYC-driven carcinogenic transcriptional programs. Overall, LL-K8-22 is the first-in-class degrader of cyclin C and would be useful for studying the unknown functions of cyclin C.

Laser-induced acoustic desorption coupled with electrospray ionization mass spectrometry for rapid qualitative and quantitative analysis of glucocorticoids illegally added in creams.

We present a strategy for the coupling of laser-induced acoustic desorption (LIAD) with electrospray ionization (ESI) mass spectrometry. Different from desorption electrospray ionization (DESI) or paper spray ionization (PSI), the technique decouples the desorption of analytes from the subsequent ionization. The desorption is initiated by a shock wave induced in 10 µm titanium (Ti) foil coated with the sample, irradiated from the rear side by a laser beam, and then the desorbed neutral analytes are post-ionized by ESI and finally characterized by quadrupole/time-of-flight (Q-TOF) mass spectrometry (MS). Separating desorption from the ionization event makes this technique flexible and decreases the matrix effect and salt effect. Various kinds of common creams containing glucocorticoids are investigated using LIAD/ESI/MS without sample pretreatment. The results show that volatile and nonvolatile analytes in creams are sampled simultaneously by LIAD, providing a convenient way for high-throughput screening of the target compounds. In addition, quantitation of glucocorticoids in creams was performed by analyzing samples with decreasing concentrations of analytes (dexamethasone (20 µg g-1) used as an internal standard (IS)), until no more signal was observed. The limits of detection (LODs) of glucocorticoids were determined experimentally to be ranging from 0.7 µg g-1 for triamcinolone acetonide to 10 µg g-1 for beclomethasone dipropionate, which are two orders of magnitude lower than the regular usage of glucocorticoids (beclomethasone dipropionate 0.25 mg g-1, triamcinolone acetonide 0.25 mg g-1). Overall, LIAD/ESI/MS is demonstrated to be of great practical importance for rapid qualitative and quantitative analysis of glucocorticoids in creams, and good sensitivity can be achieved without tedious sample pretreatment and time-consuming chromatographic separation, irrespective of the presence of complex matrices.

Metal Probe Microextraction Coupled to Dielectric Barrier Discharge Ionization-Mass Spectrometry for Detecting Drug Residues in Organisms.

In this study, a novel method for the direct coupling of metal probe microextraction (MPME) and a dielectric barrier discharge ionization (DBDI) source with mass spectrometry (MS) is reported. Analytes adsorbed on a tungsten needle were directly transferred to the DBDI source via rapid thermal desorption, which resulted in a limit of detection as low as 8 pg/mL. This is in part due to the "active capillary" configuration of the plasma ion source, where the efficiency of ion transfer to the MS is ∼100%. Specialty gases to maintain the plasma and carry analytes to the MS are not required. In contrast to direct one-step ionization of molecular adsorbates, the complete separation of the analyte desorption from the probe and the ionization event in our experimental setup greatly enhanced the sensitivity and detection reproducibility (RSD of 8.3%). We show detection of pyrimethamine, a first-line drug for the treatment and prevention of Plasmodium falciparum malaria all over the world, by this MPME/DBDI/MS method. The detection of drug residues in live fish and paramecium was achieved without the need for any sample pretreatment. The relative concentration of the drug in different organs of the fish was determined. This simple and convenient method has the potential for the analysis of chemicals even in single-cell organisms.

In situ Raman study of the photoinduced behavior of dye molecules on TiO2(hkl) single crystal surfaces.

In dye-sensitized solar cells (DSSCs), the TiO2/dye interface significantly affects photovoltaic performance. However, the adsorption and photoinduced behavior of dye molecules on the TiO2 substrate remains unclear. Herein, shell-isolated nanoparticle-enhanced Raman spectroscopy (SHINERS) was used to study the adsorption and photoinduced behavior of dye (N719) molecules on different TiO2(hkl) surfaces. On TiO2(001) and TiO2(110) surfaces, the in situ SHINERS and mass spectrometry results indicate S[double bond, length as m-dash]C bond cleavage in the anchoring groups of adsorbed N719, whereas negligible bond cleavage occurs on the TiO2(111) surface. Furthermore, DFT calculations show the stability of the S[double bond, length as m-dash]C anchoring group on three TiO2(hkl) surfaces in the order TiO2(001) < TiO2(110) < TiO2(111), which correlated well with the observed photocatalytic activities. This work reveals the photoactivity of different TiO2(hkl) surface structures and can help with the rational design of DSSCs. Thus, this strategy can be applied to real-time probing of photoinduced processes on semiconductor single crystal surfaces.

Gold-catalyzed intermolecular oxidation of o-alkynylbiaryls: an easy and practical access to functionalized fluorenes.

A novel gold-catalyzed intermolecular oxidation of o-alkynylbiaryls has been developed. A variety of functionalized fluorenes are readily accessed by utilizing this non-diazo approach, thus providing a viable alternative to synthetically useful fluorenes.