As a novel Rho-associated protein kinase (ROCK) inhibitor, SAR407899 exhibits significant efficacy in the fields of antihypertension, vasodilation, and improvement of erectile function. Due to its potent bioactivity and unique fragment, SAR407899 is widely recognized as an important chemical tool in the development of novel ROCK inhibitors. Until now, only one synthetic method has been reported, which exhibits several drawbacks such as high-temperature reaction and usage of explosive and toxic materials. Consequently, it is of great significance to optimize the synthetic process of SAR407899 . In this paper, we report an alternative, safe, and cost-efficient protocol for the synthesis of the ROCK inhibitor SAR407899 , which obviates the use of hazardous and costly reagents. Developed in innovative research training courses, this method exemplifies the successful integration of research-oriented education with practical synthetic chemistry. The application of a complete synthetic process of a drug in undergraduate laboratory courses represents an exploration of experimental teaching in college. The results verified that the teaching approach is suitable for undergraduate experiment courses. It not only developed the students’ experimental skills but also motivated their research interest.

The epidermal growth factor receptor (EGFR) represents one of the most promising targets for non-small cell lung cancer (NSCLC) therapy. However, the clinical efficacy of EGFR inhibitors is often constrained by drug resistance. EGFR resistance is primarily caused by amino acid mutations in the receptor, with common mutations such as T790M, L858R and C797S. To deeply understand the mechanism of EGFR resistance, in the present work, we employed molecular dynamics (MD) simulation to examine the interaction between the wild-type and several mutant EGFRs (including EGFRL858R, EGFRT797S, EGFRT790M/L858R, EGFRT797S/C797S and EGFRT797S/L858R/C797S) and the known EGFR inhibitors (Gefitinib, Osimertinib and TQB3804). The present work demonstrates that TQB3804 exhibits strong affinity and inhibition effects against various mutant forms of EGFR, indicating its potential to overcome resistance. Moreover, computational analyses of HOMO-LUMO energy gap revealed that TQB3804 interacts more stably at the EGFR active site than Osimertinib. The present work provides a crucial theoretical foundation for understanding the mechanisms of EGFR inhibitors and their resistance, offering valuable guidance for future drug design.

Afuresertib is one of the only two AKT inhibitors currently undergoing critical clinical development. The current preparations for afuresertib are limited to suboptimal overall yields and the use of hazardous chemical reagents, thereby emphasizing the need for a novel preparation method for bulk synthesis. This study provides a novel synthesis process for afuresertib that is characterized by environmentally friendly reaction conditions and improved overall yield. In this process, the starting material 4-bromothiophene-2-carboxaldehyde (17) was subjected to four reactions of chlorination, Suzuki coupling, secondary chlorination and oxidation to give the 2-thiophencarboxylic acid intermediate (5). While intermediate 10 was synthesized via four reactions including reduction, Boc protection, substitution reaction and deprotection using (S)-2-amino-3-(3-fluorophenyl)propanoic acid (6) as the starting material. Finally, the final product afuresertib in 99.6

Drug resistance seriously affects the treatment effect and survival rate of colorectal cancer (CRC) patients. The discovery of novel drugs and mechanisms is an important way to overcome drug resistance. Here, Tubocapsenolide A (TA), a major withanolide isolated from Tubocapsicum anomalum, significantly inhibited the growth of patient-derived organoids with oxaliplatin or 5-fluorouracil resistance and HCT116-/DLD-1-derived xenografts. Leveraging Proteolysis Targeting Chimera (PROTAC)-based target identification approach, PLK1 was identified as a direct target of TA. Mechanistically, PLK1 was first determined as a p53 cytoplasmic anchoring factor to prevent p53 from inducing ferroptosis in CRC. TA competitively inhibits the formation of PLK1-p53 heterodimers, promotes the nuclear translocation of p53, and then activates ferroptosis. Collectively, our study clarified the key role of PLK1 in p53-mediated ferroptosis for the first time and elucidated a novel mechanism of TA as a natural ferroptosis inducer in CRC treatment. The activation of the PLK1-p53-ferroptosis signaling axis may provide a brand-new strategy for the treatment of CRC.

One of the main challenges in small molecule drug discovery is finding novel chemical compounds with desirable activity. Traditional drug development typically begins with target selection, but the correlation between targets and disease remains to be further investigated, and drugs designed based on targets may not always have the desired drug efficacy. The emergence of machine learning provides a powerful tool to overcome the challenge. Herein, a machine learning-based strategy is developed for de novo generation of novel compounds with drug efficacy termed DTLS (Deep Transfer Learning-based Strategy) by using dataset of disease-direct-related activity as input. DTLS is applied in two kinds of disease: colorectal cancer (CRC) and Alzheimer's disease (AD). In each case, novel compound is discovered and identified in in vitro and in vivo disease models. Their mechanism of actionis further explored. The experimental results reveal that DTLS can not only realize the generation and identification of novel compounds with drug efficacy but also has the advantage of identifying compounds by focusing on protein targets to facilitate the mechanism study. This work highlights the significant impact of machine learning on the design of novel compounds with drug efficacy, which provides a powerful new approach to drug discovery.

The novel coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread worldwide. The main protease (Mpro) of SARS-CoV-2 plays a central role in viral replication and transcription and represents an attractive drug target for fighting COVID-19. Many SARS-CoV-2 Mpro inhibitors have been reported, including covalent and noncovalent inhibitors. The SARS-CoV-2 Mpro inhibitor PF-07321332 (Nirmatrelvir) designed by Pfizer has been put on the market. This paper briefly introduces the structural characteristics of SARS-CoV-2 Mpro and summarizes the research progress of SARS-CoV-2 Mpro inhibitors from the aspects of drug repurposing and drug design. These information will provide a basis for the drug development of treating the infection of SARS-CoV-2 and even other coronaviruses in the future.

Human immunodeficiency virus type 1 (HIV-1) is characterized by high variability and drug resistance. This has necessitated the development of antivirals with a new chemotype and therapy. We previously identified an artificial peptide with non-native protein sequence, AP3, with the potential to inhibit HIV-1 fusion through targeting hydrophobic grooves on the N-terminal heptad repeat trimer of viral glycoprotein gp41. Here, a small-molecule HIV-1 inhibitor targeting chemokine coreceptor CCR5 on the host cell was integrated into the AP3 peptide, producing a novel dual-target inhibitor with improved activity against multiple HIV-1 strains including those resistant to the currently used anti-HIV-1 drug enfuvirtide. Its superior antiviral potency in comparison with the respective pharmacophoric moieties is in consonance with the dual binding of viral gp41 and host factor CCR5. Therefore, our work provides a potent artificial peptide-based bifunctional HIV-1 entry inhibitor and highlights the multitarget-directed ligands approach in the development of novel therapeutic anti-HIV-1 agents.

Reprogramming of energy metabolism is one of the basic characteristics of cancer and has been proved to be an important cancer treatment strategy. Isocitrate dehydrogenases (IDHs) are a class of key proteins in energy metabolism, including IDH1, IDH2, and IDH3, which are involved in the oxidative decarboxylation of isocitrate to yield α-ketoglutarate (α-KG). Mutants of IDH1 or IDH2 can produce d-2-hydroxyglutarate (D-2HG) with α-KG as the substrate, and then mediate the occurrence and development of cancer. At present, no IDH3 mutation has been reported. The results of pan-cancer research showed that IDH1 has a higher mutation frequency and involves more cancer types than IDH2, implying IDH1 as a promising anti-cancer target. Therefore, in this review, we summarized the regulatory mechanisms of IDH1 on cancer from four aspects: metabolic reprogramming, epigenetics, immune microenvironment, and phenotypic changes, which will provide guidance for the understanding of IDH1 and exploring leading-edge targeted treatment strategies. In addition, we also reviewed available IDH1 inhibitors so far. The detailed clinical trial results and diverse structures of preclinical candidates illustrated here will provide a deep insight into the research for the treatment of IDH1-related cancers.

Lymphoma and leukemia have been the top two hematopoietic cancers in US, leading to more than 40,000 deaths each year. Although the 5-year relative survival rates have been improved up to 60%, there is still a lot of patients showed not response to standard therapy. Patient-Derived Xenograft model (PDX model), as the classical mouse xenograft tumor model that best represents the genetic information characteristics of human tumors, can be used to predict the therapeutic effect, and develop individualized treatment for patients. Its greatest advantage is that the model retains the microenvironment of the original tumor cells, inherits all the molecular biological properties of the primary tumor, and preserves the heterogeneity of the tumor. However, due to the factors that the loss of tumor heterogeneity in hematopoietic cancers after ex vivo clonal expansion and selection, and this in vitro assay based system could not fully reflect the host response to the drugs, including a range of novel immunotherapeutic agents such as PD-1 monoclonal antibody, CD38 monoclonal antibody, etc. With the increasing clinical use of oncology immunological agents, recently we are developing a new assay, named IO-FIVE (Immuno-Oncology drugs Fast In Vivo Efficacy test) for research and therapy, by retaining an appropriate proportion of tumor-infiltrating immune cells (and stromal cells) mixed with tumor cells when digesting clinical tumor samples or patient-derived xenografts-preserved tumor samples into cell suspensions, and then injecting them into the specialized IO-FIVE device, a modified microencapsulation and hollow fiber culture system (OncoVee® MiniPDX), which is implanted subcutaneously into mice and administered systematically for immuno-drug susceptibility testing to screen for immuno-drugs or combinations suitable for individual treatment. The IO-FIVE testing cycle takes only 14 days and is performed using Celltiter Glo before and after the injection of cell suspensions into the specialized device. Celltiter Glo, flow cytometry and Omics (RNA-seq, DNA-seq) are used to measure the viability of total cell subpopulations, the relative ratio of tumor cells to immune cells and the alteration of cellular transcription levels in the device, in order to further explore the sensitivity of tumor tissues to the immunotherapeutic drugs and the potential molecular mechanism for the responders and non-responders. Immune regulatory CD38 antibody Daratumumab has been tested in more than 40 acute myeloid leukemia (AML) cases, with its overall in vivo efficacy around 25%, which is consistent with previous research. It is interesting to uncover novel pathways for drug discovery and clinic treatment since CD38 is well expressed in most AML patients, but at least half of those CD38 + AML patients are not sensitive or resistant to Daratumumab monotherapy. Expression of CD38 on AML is not enough to distinguish the responder vs non-responder of Daratumumab. IO-FIVE could potentially be a companion diagnosis on AML for the patient stratification of Daratumumab. PD1 antibody Sintilimab has been tested in all the patients (12 lymphoma, 52 leukemia), and the only two laboratory PD1-Ab non-responsive T cell lymphoma patients have been showed to be clinically non-responders as well. Immunotherapy and chemical or targeted therapy combination could enhance tumor killing ability and disease control. We will use the IO-FIVE data to further compare it with its source of real-world clinical patients to obtain more evidence-based evidence that can predict individualized treatment with immunotherapy.

CD73, as a surface enzyme anchored on the outside of the cell membrane via glycosylphosphatidylinositol (GPI), can convert the AMP in the tumor cell microenvironment into adenosine to promote the growth of tumor cells. It has been overexpressed in many different types of human tumors, such as gastric cancer, pancreatic cancer, liver cancer, and other tumor cells. Therefore, targeted inhibitors of CD73 are considered potential tumor treatment methods. Due to the low bioavailability of nucleoside CD73 inhibitors, it is necessary to develop new inhibitors. In this study, through molecular docking, three-dimensional quantitative structure–activity relationship (3D-QSAR) and molecular dynamics (MD) simulations, a series of CD73 inhibitors were calculated and studied to reveal their structure–activity relationships. Through molecular docking studies, the possible mode of interaction between inhibitors and protein is explored. Subsequently, a 3D-QSAR model was established by comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA). For the best CoMFA model, the Q2 and R2 values are 0.708 and 0.983, respectively, while for the best CoMSIA model, the Q2 and R2 values are 0.809 and 0.992, respectively. Based on the contour maps, we designed ten new CD73 inhibitors and predicted their activity by the model, all of them are better than molecules in the dataset. In addition, in order to select potential drug candidates, ADMET prediction was performed on template molecules and designed compounds. Moreover, the stability of the complex formed by the two inhibitors and CD73 was evaluated by molecular dynamics simulation, and the results are consistent with the results of molecular docking and 3D-QSAR research. Finally, the binding free energy was calculated by the surface area method (MM-GBSA), and the results are consistent with the activities that van der Waals and Coulomb contribute the most during the binding process of the molecule to the CD73 protein. In conclusion, our research provides valuable information for the further development of CD73 inhibitors.

Molecular imprinting polymer (MIP) has been increasingly employed for sulfonylurea herbicides (SUHs) detection in different matrices. A novel MIP that was effective as a highly class-selective sorbent in molecularly imprinted solid-phase extraction (MISPE) was successfully prepared for isolation and purification of SUHs, namely, metsulfuron-methyl, chlorsulfuron, chlorimuron-ethyl, prosulfuron, and pyrazosulfuron-ethyl, in rice, corn and soybean samples. The MIP was synthesized by precipitation polymerization using metsulfuron-methyl as the template, 4-vinylpyridine as the functional monomer, ethylene glycol dimethacrylate as the crosslinker, and MeCN as the porogen. The polymerization system of the MIP was optimized, and its adsorption performances were evaluated by comparing its adsorption isotherms and adsorption kinetics with those of a non-imprinted polymer (NIP). Following MISPE for extracting and enriching SUHs from rice, corn and soybean samples, high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was performed. Acceptable recoveries were observed at SUHs contaminant concentrations of 10, 20 and 40 μg/L: from 77.56 to 99.81%, with relative standard deviations of <13.8% (n = 5) for all samples. The limits of detection for the five SUHs were 0.21-0.26 μg/L. The results demonstrated that the proposed MISPE-HPLC-MS/MS method is an effective approach for the simultaneous and sensitive determination of the five SUHs in rice, corn and soybean samples.

Nonsmall cell lung cancer (NSCLC) is one of the most common malignancies and needs novel and effective chemotherapy. In this study, our purpose is to explore the anticancer effects of 2-methoxy-5((3,4,5-trimethosyphenyl) seleninyl) phenol (SQ) on human NSCLC (A549 and H460) cells. We found that SQ suppressed the proliferation of NSCLC cells in time- and dose-dependent manners, and blocked the cells at G2/M phase, which was relevant to microtubule depolymerization. Additionally, SQ induced A549 and H460 cell apoptosis by activating the mitochondrial apoptotic pathway. Further, we demonstrated that SQ enhanced the generation of reactive oxygen species (ROS), and pretreatment with N-acetyl- L-cysteine (NAC) attenuated SQ-induced cell apoptosis. Meanwhile, SQ mediated-ROS generation caused DNA damage in A549 and H460 cells. Our data also revealed that SQ-induced apoptosis was correlated with the inhibition of mouse double minute 2 (MDM2) in A549 and H460 cells. In summary, our research indicates that the novel compound SQ has great potential for therapeutic treatment of NSCLC in future.

The accumulation of β-amyloid (Aβ) in the brain plays an important role in the pathogenesis of Alzheimer’s disease (AD). The lack of estrogen is one of the risk factors for AD. Quercetin is a phytoestrogen with a chemical structure similar to that of estrogen. However, the mechanism by which quercetin prevents AD is unclear. PC12 cells were cultured with Aβ25–35 for 24 h. Then the cells were further treated with 17β-estradiol, genistein, and quercetin for another 24 h, respectively. Next, ICI182780 and U0126 were used to study the mechanisms of estrogen-like neuroprotection. Methyl thiazolyl tetrazolium (MTT) assay was performed to detect cell survival. The protein expression was analyzed by immunofluorescence and western blot. The survival of PC12 cells induced by Aβ25–35 was increased by quercetin. The levels of estrogen receptor α (ERα) and p-extracellular signal-regulated kinase (ERK)1/2 were improved by quercetin, but not those of ERβ. On the contrary, Bcl-2/Bax was increased and the expression of Caspase-3 was decreased. When the cell was pretreated with ICI182780, the p-ERK1/2 and Bcl-2/Bax ratio was decreased, but Caspase-3 expression was increased. In addition, pretreatment with U0126 would reduce Bcl-2/Bax ratio and increase Caspase-3 protein expression. Conclusively, quercetin plays a neuroprotective role through the ER pathway and the mitogen-activated protein kinase (MAPK) pathway. The MAPK signaling pathways could also be activated by quercetin via the mediation of ERα.

Nanocrystals (NCs) exhibit potential in improving oral bioavailability for poorly water-soluble drugs. However, whether NCs improve oral absorption by quick dissolution or by endocytosis remains inconclusive because tracking of dissolved drugs and NCs particles cannot occur simultaneously. In this study, we aim to elucidate how NCs improve oral absorption by using coumarin 6 (C6), an aggregation-caused quenching fluorophore, and 2-((5-(4-(dip-tolylamino)phenyl)thiophen-2-yl)methylene)malononitrile (MeTTMN), an aggregation-induced emission fluorophore. C6 was used as a model drug to prepare NCs and MeTTMN was incorporated to construct fluorescence resonance energy transfer (FRET) pairs. Thus, the molecular absorption can be detected using the fluorescence signal of dissolved C6 and the NCs particles can be tracked simultaneously by monitoring FRET signals. The reliability of this tracking method was validated. Accordingly, in vitro dissolution, gastrointestinal traffic, and biodistribution studies were conducted. The results showed that dissolved C6 molecules were the main absorption mode of C6 NCs. Identification of such pathways bears considerable significance for the broad application of drug NCs in improving the druggability of insoluble drugs.

Natural products are one of the important sources for the discovery of new drugs. Betulinic acid (BA), a pentacyclic triterpenoid widely distributed in the plant kingdom, exhibits powerful biological effects, including antitumor activity against various types of cancer cells. A considerable number of BA derivatives have been designed and prepared to remove their disadvantages, such as poor water solubility and low bioavailability. This review summarizes the current studies of the structural diversity of antitumor BA derivatives within the last five years, which provides prospects for further research on the structural modification of betulinic acid.

Fluorescence imaging is a promising visualization tool and possesses the advantages of in situ response and facile operation; thus, it is widely exploited for bioassays. However, traditional fluorophores suffer from concentration limits because they are always quenched when they aggregate, which impedes applications, especially for trace analysis and real-time monitoring. Recently, novel molecules with aggregation-induced emission (AIE) characteristics were developed to solve the problems encountered when using traditional organic dyes, because these new molecules exhibit weak or even no fluorescence when they are in free movement states but emit intensely upon the restriction of intramolecular motions. Inspired by the excellent performances of AIE molecules, a substantial number of AIE-based probes have been designed, synthesized, and applied to various fields to fulfill diverse detection tasks. According to numerous experiments, AIE probes are more practical than traditional fluorescent probes, especially when used in bioassays. To bridge bioimaging and materials engineering, this review provides a comprehensive understanding of the development of AIE bioprobes. It begins with a summary of mechanisms of the AIE phenomenon. Then, the strategies to realize accurate detection using AIE probes are discussed. In addition, typical examples of AIE-active materials applied in diagnosis, treatment, and nanocarrier tracking are presented. In addition, some challenges are put forward to inspire more ideas in the promising field of AIE-active materials.

The discovery of targetable mutations, which cause gene rearrangement, led to a major advancement in the treatment of patients with non-small cell lung cancer (NSCLC), and cancers with such mutations can be paired with drugs which specifically target them. c-ros oncogene (ROS1) positive NSCLC is one molecular subtype of NSCLC with a therapeutic target. Currently, different targeted therapies and ROS1 inhibitors have been discovered, but all are in different investigational phases, with only one (crizotinib) which is FDA approved. Crizotinib is a small-molecule tyrosine kinase inhibitor (TKI) which was discovered to actively inhibit ALK, MET, and ROS1. Crizotinib has shown to be remarkably efficacious against ROS1 lung cancer, prompting ROS1 detection in lung cancer to be quite significant. Sadly, crizotinib resistance in ROS1 is a frequent occurrence which poses a major clinical challenge in the successful treatment of ROS1 lung cancer; hence, the discovery of the second and third generation ROS1 inhibitors is of utmost importance. In this review, we discuss the underlying mechanisms through which ROS1 tumor cells acquire resistance to crizotinib—the first-line drug for ROS1-positive NSCLC, and summarize various new potent drugs which can overcome this resistance and serve as viable alternatives.

Description of the back cover image: The cover image is based on the REVIEW ARTICLE A new approach to developing diagnostics and therapeutics: aggregation-induced emission based fluorescence turn-on by Meichen Guo et al., https://doi.org/10.1002/med.21595.

2-Methoxy-5((3,4,5-trimethosyphenyl)seleninyl) phenol (SQ), a novel synthesized combretastatin A-4\n(CA-4) analogue, is identified as a microtubule inhibitor and has been shown to exert anticancer activity in breast cancer cells. Here, we found that SQ reversed epidermal growth factor (EGF)-induced motility and invasion in breast cancer cell lines by the in vitro Wound healing and Transwell assay. Further studies showed that SQ treatment resulted in inhibitory alteration of EGF-stimulated epithelial-to-mesenchymal transition (EMT) and MMP-2 activity. What is more, SQ significantly inhibited the EGF-induced mouse double minute 2- (MDM2) expression and transcription factor Twist1 expression. In addition, compared with the control cells, MDM2 overexpression up-regulated Twist1 expression and dramatically promoted cell migration and invasion, MDM2 under-expression also down-regulated Twist1 expression and suppressed cell motility and invasion. Taken together, our findings suggest that the inhibitory effects of SQ on migration and invasion were related to the suppression of MDM2 and Twist1 signal axis.

It is still challenging to determine the potential targets of natural products, which is essential for further drug research and development. Due to its novel mechanism of action of inducing autophagy effects in breast cancer cells, asperphenamate has received our considerable attention. However, its unknown target inevitably impedes further study. In our previous work, the target enzyme of asperphenamate was predicted as cathepsin by the natural product consensus pharmacophore strategy. Then, asperphenamate and its three derivatives were chosen to study in detail by molecular docking calculations with AutoDock 4 suite. The docking results showed the three derivatives interacted more tightly with either cathepsin L or cathepsin S than with asperphenamate. The ortho-benzyloxyl phenylacetyl derivative 1 andp-toluenesulfonyl derivative 3 showed similar interactions with cathepsin L and adopted a better geometric shape within the binding pocket than did the N-CBZ-piperidyl analog 2. On the other hand, compound 2 formed more hydrogen bonds than 1 and 3 to make it tightly bind within cathepsin S. The cathepsin inhibitory activity assay verified the molecular simulation results. Compound 2 was remarkably less active than 1 and 3 against cathepsin L. However, compound 2 showed the strongest potency against cathepsin S with IC50 of 13.12 ± 0.29 μM. Considering that cathepsin S plays a vital role in the process of metastasis in breast cancer cells, the inhibitory effect of 2 on migration and invasion was further studied in human breast cancer MDA-MB-231 cells by wound healing and transwell chamber assays. The results illustrated that 2 exhibited an apparent inhibitory ability to the metastasis of MDA-MB-231 cells. Next, 2 will be chosen as a lead compound to develop novel double functional chemotherapeutic agents with both novel mechanisms of action against apoptosis-resistant cancer cells, such as inducing autophagy and inhibiting cancer metastasis.