Recombinant human granulocyte colony stimulating factor (rhG-CSF) has been in clinical use for the adjuvant therapy of cancer patients with neutropenia caused by radiotherapy/chemotherapy. However, it does have some drawbacks such as poor stability and short half-life, and needs to be administered repeatedly, which is easy to cause adverse reactions such as drug tolerance and immune rejection. Therefore, it is necessary to develop long-acting rhG-CSF to improve its clinical efficacy. In this review, we summarize the research progress on the development of long-acting rhG-CSF using the strategies such as PEGylation, fusion protein and new dosage forms in recent years, and discuss its future development trend.

This study investigated the effect of morin, an active ingredient of the family Moraceae, on collagen-induced arthritis (CIA) in mice, and explored the underlying mechanism from the perspective of recovering immune balance. The collagen was used to induce model of CIA in mice, morin was administered by gavage, and arthritis index (AI) score, imaging and histopathological changes of the paws and ankle joints, and the levels of proinflammatory factors, proinflammatory mediators as well as the IgG class antibodies in serum were detected. In addition, the frequencies of T helper 17 (Th17) and regulatory T (Treg) cells and the levels of relevant transcription factors and functional factors in lymph nodes/spleen as well as the levels of interleukin-17A (IL-17A) and IL-10 in serum were determined. The results showed that oral administration of morin significantly reduced the AI score, improved joint swelling and bone damage, reduced the pathological score, and down-regulated the levels of proinflammatory factors [tumor necrosis factor-‍α (TNF-‍α), IL-6 and IL-1β], proinflammatory mediators [prostaglandin E₂ (PGE₂), matrix metalloproteinase-13 (MMP-13) and nitric oxide (NO)] and IgG class antibodies (IgG and IgG2a) in serum. Moreover, the percentage of Th17 cells, the expressions of Th17-specific transcription factor retinoic acid-related orphan receptor γt (RORγt) and functional factors IL-17A, IL-21 and IL-22 in lymph nodes/spleen, as well as the level of IL-17A in serum were down-regulated, while the percentage of Treg cells, the expressions of Treg-specific transcription factor forkhead box P3 (Foxp3) and functional factors IL-10 and transforming growth factor-‍β (TGF-‍β) in lymph nodes/spleen, as well as the level of IL-10 in serum were up-regulated. All animal treatments were approved by the Animal Ethics Committee of China Pharmaceutical University and strictly followed the welfare regulations of laboratory animals of China Pharmaceutical University. This study indicates the therapeutic effect of morin on mice with CIA, and the mechanism is associated with the improvement of Th17/Treg imbalance, which provides a theoretical basis for the clinical application of morin.

To explore the protective effects and pharmacophore of verbascoside against oxygen-glucose deprivation/reperfusion (OGD/R)-induced neuronal cells injury, we used OGD/R-induced PC12 cells as a neuronal injury model. We investigated the neuroprotective effects of verbascoside and its structural fragments (caffeic acid 3, 4-dihydroxyphenethyl ester, caffeic acid, and 3, 4-dihydroxyphenylethanol) by MTT and crystal violet staining analysis. Moreover, we studied the protection of verbascoside and its structural fragments on mitochondria by Hoechst33258 staining, JC-1 staining and transmission electron microscope analysis. The neuroprotective mechanisms of verbascoside and its major active fragment caffeic acid were investigated by detecting B cell lymphoma 2 (Bcl 2)/Bcl 2 associated X protein (Bax)-dependent mitochondrial cysteinyl aspartate specific proteinase 3 (caspase 3)/poly ADP-ribose polymerase (PARP) apoptosis pathway by Western blot. The results showed that verbascoside, caffeic acid 3, 4-dihydroxyphenethyl ester and caffeic acid significantly improved cell viability and maintained normal PC12 cells morphology. These compounds also significantly reversed OGD/R-induced PC12 cells apoptosis, inhibited cell mitochondria depolarization, and maintained normal mitochondria structure. Furthermore, verbascoside and its major active fragment caffeic acid markedly inhibited the cleavage of mitochondrial apoptotic proteins caspase 3 and PARP, down-regulated Bax, and increased Bcl 2 expression. These results indicate that verbascoside protects OGD/R-induced neuronal cells injury via mitochondrial caspase 3/PARP apoptosis pathway, and caffeic acid may function as the major pharmacophore structure.

Viral infections pose a persistent threat to human health and life. The rapid emergence of drug-resistant strains and the outbreak of new viruses require researchers to develop innovative strategies to accelerate the development of more antiviral drugs towards novel targets to meet the clinical needs. This paper selects typical research cases and reviews the novel targets and strategies of antiviral drugs in recent years from the perspective of medicinal chemistry.

The success of new drug discovery in 2021 can be affirmative, although the whole world is still suffering from COVID-19 pandemic. 50 new drugs were approved by the FDA's Center for Drug Evaluation and Research (CDER) last year. Among them, 27 were defined as first-in-class drugs, accounting for the highest number in the past decade. Notably, small molecule drugs still occupy a dominant position in first-in-class drugs with 15 drugs approved. Some of them were regarded as milestones for the drug discovery including sotorasib, a first small molecular covalent inhibitor targeting the "undruggable" target of the KRAS G12C; asciminib, a first small molecular allosteric inhibitor targeting the allosteric pocket of BCR-ABL1; belzutifan, a first small molecular inhibitor to inhibit HIF-2α; and vericiguat, a first small molecular sGC agonist for the treatment of chronic heart failure (CHF). First-in-class drugs rely on the discovery of novel targets and biological mechanisms, thus requiring different drug design approaches and being important guidance. In this review, we expect to provide research ideas and methods for more first-in-class drugs based on the research background, development process and therapeutic application of 3 first-in-class small molecule drugs in 2021.

Using Lonicera japonica genomic DNA as a template, we cloned Lonicera japonica U6 promoters. Four LjU6 promoters, 336, 708, 359 and 602 bp in length, were cloned by PCR from Lonicera japonica genomic DNA. PlantCARE analysis found that the four promoters contained typical promoter cis-elements, such as a TATA box and CAAT box, and contained regulatory elements related to light response and stress response. After the cloning products were sequenced, the LjU6 promoter was ligated to the pBI121 vector carrying the β-glucuronidase (GUS) gene to construct four LjU6-pBI121 fusion expression vectors. Nicotiana tabacum leaves were transformed by the Agrobacterium transient transformation method and GUS histochemical staining was performed on the leaves. The staining results showed that LjU61-F1 had the highest transcriptional activity. This study thus identified a U6 promoter with high transcriptional activity, providing a basis for the establishment of CRISPR/Cas9 genome editing technology in Lonicera japonica.

This study aimed to research the antitumor activity and mechanisms of caffeic acid phenethyl ester derivative PEC01 in mouse G422 glioma. MTT assay, flow cytometry (FCM) and Transwell migration assay were used to detect the effects of PEC01 on proliferation, apoptosis, and migration of G422 cells respectively. Mouse subcutaneously transplanted G422 tumor model was used to analyse the effect of PEC01 on the growth of glioma in vivo. Animal welfare and experimental procedure are in accordance with the regulations of the Animal Ethics Committee of institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College. Western blot was used to detect the protein levels of epidermal growth factor receptor (EGFR), Src and their downstream signaling pathways in G422 cells and tumor issue. The results showed that PEC01 inhibited proliferation of G422 cells in a time- and dose-dependent manner, with IC₅₀ of (9.02 ± 0.36) μmol·L^-1 at 96 h. PEC01 significantly induced early apoptosis and late apoptosis of G422 cells at 10.0 and 20.0 μmol·L^-1 concentrations for 96 h. Scratch healing rate of G422 cells reduced after treated with different concentrations (0.625-5.0 μmol·L^-1) of PEC01 for 12-48 h in scratch healing assay. The number of transmembrane G422 cells decreased in groups treated with PEC01 for 8 h compared with DMSO group. The average tumor weight of groups treated with 30.0 and 60.0 mg·kg^-1 PEC01 was significantly reduced in G422 insubcutaneously transplanted tumor model, and the inhibition rate of tumor weight was 72.29% and 59.44%, respectively. Protein levels of EGFR, Src, c-myc and hypoxia-inducible factor 1-alpha (HIF-1α) decreased significantly in G422 cells and tumor tissue. The mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway and phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway related proteins were down-regulated. Related proteins of invasion, metastasis and cell cyclin were significantly down-regulated. PEC01 can suppress the growth of G422 glioma in vitro and in vivo. The antitumor activity of PEC01 in mouse subcutaneously transplanted G422 tumor model might be related to the blockcade of PI3K/Akt/mTOR and MAPK/ERK signaling pathways.

Polysaccharides as one of the most common drug delivery materials have the excellent advantages, such as diverse natural sources, high biocompatibility and multi-functions. Polysaccharides have been investigated and widely used in food industry, pharmaceutical and medical fields especially in the targeted oral drug delivery for colonic diseases treatment with important research values and great potential applications. Inspired by the unique properties of polysaccharides, researchers around the world have developed the cross-linked nanoparticles, self-assembled nanoparticles and hydrogels, focusing on various drug delivery strategies such as pH-sensitive, microbial enzyme-responsive, reactive oxygen species-responsive, mucoadhesive and receptor-targeted. Exhilaratingly, the polysaccharides-based therapeutics have shown high efficacy for the treatment of digestive tract diseases, such as colitis or colonic cancer. Herein, we summarized the research progress of polysaccharides-based targeted oral drug delivery systems for colonic diseases treatment, and discussed the perspectives of the researches and application development in future.

As a kind of tumor immunotherapy, tumor vaccine provides a new strategy for cancer treatment. With nano-biomimetic materials to encapsulate the tumor antigens, the construction of nano-biomimetic tumor vaccine can target the tumor and release antigens, with high efficiency and safety. Therefore, nano-biomimetic vaccine has become a hot research topic. Based on this review, several new nano-biomimetic nanoparticles are summarized, and the clinical applications of the nano-biomimetic vaccine combined with other therapeutic strategies are introduced.

The purpose of the study was to investigate the thermal expansion characteristics of brivaracetam form Ⅰ, and explore the influence mechanism of the crystal structure on its thermal expansion behavior. The crystal structure of brivaracetam form Ⅰ was characterized by X-ray single crystal diffraction and variable temperature X-ray powder diffraction at different temperatures. The interaction energy of brivaracetam molecule calculated by B3LYP/6-31G(d, p) wave function with the aid of CrystalExplorer 21.5 software. The results show that brivaracetam form Ⅰ exhibits significant reversible anisotropic thermal expansion under the temperature range of 123-323 K. The principal expansion X₁, X₂, X₃ axes are approximately aligned with the a, b and c axes of the unit cell, and the thermal expansion coefficients of the principal expansion axes are -127.61×10^-6, 95.96×10^-6, 233.80×10^-6 K^-1, respectively. The a-axis exhibits negative expansion characteristics. The volumetric thermal expansion coefficient is 202.17×10^-6 K^-1. The energy framework of the crystal is obvious layered, and the interaction energy between layers is weak, which leads to a significant linear positive expansion in the c-axis direction of the unit cell. Through a combination of experimental and theoretical methods, the thermal expansion characteristics of brivaracetam form Ⅰ are systematically analyzed, and the influence mechanism of the crystal structure on its thermal expansion behavior is explored, which has certain guiding significance for the production process of tablet preparations in practice.