Abstract Cardiovascular diseases (CVDs), including atherosclerosis, congestive heart failure, and hypertension, are most commonly observed in middle-aged and elderly individuals. Approximately 15 million people die from them every year worldwide. CVDs are associated with high morbidity, disability, and mortality rates, posing a significant threat to public health. Although the importance of natural medicines in treating CVDs has become increasingly prominent, there is still much to be discovered about their composition and mechanisms of action. With the rise of network pharmacology as a valuable tool to understand how natural medicines work, researchers can study the body's responses to disease stimuli and drug therapy by constructing a “component–target–pathway–disease” interaction network. In this work, we summarized the application of network pharmacology in the study of CVDs. We also discussed the challenges and opportunities by this tools, hoping to provide suggestions for the treatment of the disease in the future.

Quetiapine, a dibenzothiazepine derivative, is an antagonist of serotonin and dopamine. It was approved by the Food Drug and Administration for the treatment of schizophrenia in 1997 and has been used as a commonly prescribed second-generation atypical antipsychotic drug. Loxapine is a dibenzoxazepine tricyclic compound used as an antipsychotic, antagonising dopamine and serotonin receptors for the treatment of acute and chronic schizophrenia. However, the preparation of their Key Starting Materials (KSMs) in the reported synthesis routes suffers from several significant restrictions, such as multistep synthesis, harsh reaction conditions, high cost factors, and the use of reagents that are environmentally unfriendly. In this work, we aimed to explore a telescopic green process for the synthesis of dibenzo[b,f][1,4]thiazepin-11(10H)-one and 2-chlorodibenzo[b,f][1,4]oxazepin-11(10H)-one, the two KSMs required for the commercial production of quetiapine and loxapine. The process involves an intermolecular base-mediated SNAr reaction of 2-fluoro-1-nitrobenzene and methyl 2-mercaptobenzoate or methyl 2-hydroxybenzoate, followed by intramolecular reductive amidation employing sodium dithionite (Na2S2O4) as the sole reagent. The SNAr reaction was performed in the presence of K2CO3 in DMF at 90°C, followed by workup to give a crude product, which was treated with Na2S2O4 in DMSO without any prepurification to obtain the desired cyclized KSMs. Unlike the commonly used metal/acid reagent for the reduction of nitro compounds, the developed process avoids the use of any metal reagent or acidic conditions. The key features include a reduced number of steps, a telescopic process avoiding purification of the first step product, and reductive amidation of unactivated esters without any externally added activating agent.

In the development of dry powder inhaler (DPI) formulations, the choice and optimization of carriers are critical, as they directly impact not only drug stability and bioavailability but also patient adherence. This study sought to create and synthesize a novel DPI excipient with enhanced qualities relative to the current excipient, i.e., fumaryl diketopiperazine (FDKP). In this work, FDKP's framework was utilized to synthesize a variety of novel compounds (DBBB1–15), preserving the diketopiperazine ring and symmetrical branched chains while implementing structural alterations. The artificial intelligence software Schrödinger was employed to screen these chemicals for potential possibilities. As a result, DBBB6 was selected because of its advantageous look and physicochemical properties, including a greater pKa (reduced acidity) when compared with FDKP. The synthesis method for DBBB6 was refined, resulting in a 9.7% yield. Significantly, investigations involving rats demonstrated that DBBB6 did not induce coughing, a possible adverse effect associated with FDKP. The results indicate that DBBB6 is a viable alternative to FDKP as a DPI excipient. Its improved tolerability profile suggests a potential for reduced adverse effects. Additional studies are required to comprehensively assess its safety and efficacy for clinical application.

Abstract Vosoritide is a therapeutic peptide that promotes skeletal growth by targeting the NPR-B receptor and was approved in 2021 for the treatment of achondroplasia. However, its high production cost poses a considerable economic burden on patients, limiting its widespread use. This study aims to establish an efficient and cost-effective biosynthetic process for the production of Vosoritide using Escherichia coli. In this work, Vosoritide was expressed as inclusion bodies (IBs) in E. coli BL21 (DE3) via a gp55 fusion strategy. Downstream processes, including fermentation, HCl-induced acidic cleavage of IBs, pH-dependent protein precipitation, as well as the performing conditions of the ion-exchange chromatography and reverse-phase chromatography, were optimized. The analysis methods included sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), high-performance liquid chromatography (HPLC), and liquid chromatography-tandem mass spectrometry. Cyclic guanosine monophosphate (cGMP) assays were conducted to assess the biological activity of the protein in NIH3T3 cells. Vosoritide was obtained at 1.3 g per liter of fermentation broth with a purity exceeding 99%. The peptide's primary structure, molecular weight, and disulfide bond integrity were also confirmed through mass spectrometry and peptide mapping. The purified Vosoritide stimulated cGMP production in NIH3T3 cells with a half-maximal effective concentration (EC50) of 0.37 nmol/L. In conclusion, this study provided a scalable, high-yield production method for Vosoritide that yields Vosoritide with pharmaceutical-grade purity and activity, holding significant potential for manufacturing a cost-effective biosimilar with broader clinical accessibility.

Abstract Melanotransferrin (CD228) is a membrane glycoprotein involved in tumor growth and metastasis and is highly expressed in various solid tumors. Despite its tumor-specific nature, no antibody drugs targeting CD228 are currently available. This study aimed to develop a humanized CD228 monoclonal antibody and its antibody–drug conjugate (ADC) to evaluate in vitro cytotoxicity and in vivo antitumor efficacy against melanoma. In this work, mice were immunized with the extracellular domain of CD228, and specific antibodies were screened using hybridoma technology, followed by humanization. The humanized antibody was conjugated to monomethyl auristatin E (MMAE) with a citrulline–valine linker and purified by protein A chromatography. The drug–antibody ratio (DAR) and purity were analyzed using hydrophobic interaction chromatography and size-exclusion chromatography. CCK8 assay was performed to assess cytotoxicity against tumor cells, and antibody-dependent cell-mediated cytotoxicity (ADCC) was assessed using Jurkat cells. An A2058 melanoma xenograft model was used to examine in vivo efficacy. This work identified a humanized antibody, Ab-2F6A1, with high CD228 binding affinity, with EC50 values of 1.746 ng/mL (protein binding activity) and 83.8 ng/mL (cell binding activity). The ADC, Ab-2F6A1-VcMMAE, had an average DAR of 4.9026 and a purity of 98.24% and showed significant in vitro cytotoxicity against melanoma, breast, and colon cancer cells. Ab-2F6A1-VcMMAE has a stronger ADCC effect compared with the positive control (Ab-hI49-VcMMAE), and exhibited superior melanoma tumor inhibition in vivo. Given the above, a novel humanized anti-CD228 antibody and its ADC were successfully developed. The ADC demonstrated strong antigen binding, in vitro antitumor activity, ADCC effects, and superior in vivo melanoma inhibition, supporting CD228 as a promising therapeutic target.

Abstract Collagenase and elastase enzymes that cause the degradation of collagen and elastase have both been implicated in skin aging. Agathosma betulina is a popular plant in South Africa used in aromatherapy and folk medicine but the scientific knowledge regarding its anti-aging activity is yet to be explored. This study aimed to investigate the roles of the essential oil of A. betulina leaves on skin aging and its inhibitory effect on collagenase and elastase activities. In this work, the chemical profiling of essential oils of A. betulina leaves was explored. The antioxidant activity of essential oil of A. betulina leaves at different concentrations (0.07–10 µg/mL) was assessed using standard assay methods for 1,1-dihenyl-2-picryhydrayl (DPPH) and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS). Standard procedures for elastase and collagenase assays were also employed to determine the anti-aging skin activity of the essential oil at the same concentrations. Gas chromatography-mass spectrometric analysis showed the presence of 43 constituents with menthone (29.2%), limonene (23.7%), and pulegone (8.4%) as the major compounds. The oil expressed strong radical scavenging activity against DPPH and ABTS radicals at higher concentrations, with over 90% inhibition, particularly at 2.5, 5, and 10 µg/mL compared to the control (butylated hydroxytoluene). The oil also demonstrated stronger inhibitory activity against elastase at higher concentrations (5 and 10 µg/mL) and concentration-dependent inhibitory activity against collagenase compared to the control (ursolic acid). The essential oil of A. betulina leaves could be a potential candidate for the cosmetic industry to retard skin wrinkles and other manifestations of skin aging particularly associated with extrinsic factors.

Abstract Diclofenac sodium is a widely used non-steroidal anti-inflammatory drug with potent analgesic, anti-inflammatory, and antipyretic properties. However, its limited aqueous solubility poses challenges in pharmaceutical formulations and analytical quantification. To address this issue, various solubility enhancement techniques have been explored to improve its bioavailability and facilitate precise spectrophotometric analysis. Hydrotropic solubilization has been widely studied as an effective technique to improve the aqueous solubility of poorly water-soluble drugs. This study aimed to investigate an eco-friendly and efficient approach to enhance the solubility of diclofenac sodium, focusing on a mixed hydrotropic approach, ensuring reliable and accurate spectrophotometric estimation without the need for organic solvents. In this work, sodium citrate and N,N-dimethylurea were explored as hydrotropic agents. The selection was based on their complementary solubilization mechanisms. Sodium citrate enhances solubility through ionic interactions, whereas N,N-dimethylurea disrupts the water structure and forms hydrogen bonds with the drug. The results showed a synergistic effect of the combined use of these agents with a solubility of 63 mg/mL, which was significantly higher than that of individual use. Besides, the Fourier transform infrared (FTIR) spectroscopy confirmed hydrogen bonding when diclofenac sodium was dissolved in the hydrotropic blend, and X-ray diffraction analysis demonstrated a decrease in the crystallinity of the drug, indicating a transition to a more soluble amorphous state. The solubility studies, molecular docking, and recovery experiments confirmed the high accuracy and precision of the proposed method with percentage recoveries near 100%. In conclusion, this green and cost-effective approach eliminates the need for toxic organic solvents, making it a sustainable and scalable solution for the analysis and formulation of poorly water-soluble drugs.

Abstract Drug resistance is a looming threat to global health security, undermining the effectiveness of current treatments and increasing treatment failures. To address this challenge, it is necessary to explore innovative strategies by identifying new binding mechanisms and targeting previously undeveloped therapeutic avenues. This paper reviewed the potential of leveraging undeveloped domains to combat drug resistance and proposes a range of methodologies to accurately identify those specific targets. There is also an extensive review of the challenges associated with targeting undeveloped areas and strategies to effectively address them. In this process, the application of artificial intelligence (AI) can effectively improve the efficiency of drug design, while appropriate attention should be paid to the physicochemical and drug-like properties of pharmaceutical compounds in the realm of drug discovery. Given the above, focusing on these undeveloped areas could provide a promising pathway to address drug resistance; however, achieving this objective necessitates sustained investigative efforts and inventive approaches.

Abstract In the fields of drug synthesis and fine chemicals, the traditional kettle-type reactor with a small heat exchange area and the microreactor which is easy to clog, are not suitable for the reaction with fast reaction speed, large heat release, and high solid content. However, the continuously stirred reactor has the advantages of high mass and heat transfer efficiency, high reaction safety, and not easy to be clogged, and therefore, has a great potential for application. This paper reviews the research progress of continuous agitation reactors in organic synthesis, drug crystallization, and biocatalysis, hoping to provide a reference for applications of continuous agitation reactors in the pharmaceutical and chemical fields.