Background: Supersaturating drug delivery systems (SDDSs) have gained significant attention as a promising strategy to enhance the solubility and bioabsorption of Biopharmaceutics Classification System (BCS) II drugs. To overcome challenges associated with polymer-based amorphous SDDS (aSDDS), coamorphous (CAM) systems have emerged as a viable alternative. Among them, “drug-drug” CAM (ddCAM) systems show considerable potential for combination drug therapy. However, many drugs in their pure amorphous forms are unstable at room temperature (RT), complicating their formation and long-term stability profiles. Consequently, limited knowledge exists regarding the behavior of ddCAMs containing RT-unstable components formed via quench cooling. Methods: In this study, we used naproxen (NAP), a RT-unstable amorphous drug, in combination with felodipine (FEL) or nitrendipine (NTP), two RT-stable amorphous drugs, to create “FEL-NAP” and “NTP-NAP” ddCAM pairs via quench cooling. Our work used a series of methods to perform a detailed analysis on the co-amorphization, dissolution, solubility, and stability profiles of ddCAMs containing RT-unstable drugs, contributing to advancements in co-amorphization techniques for generating SDDS. Results: This study revealed that the co-amorphization and stability profiles of ddCAMs containing RT-unstable components produced via a quench-cooling method were closely related to drug-drug pairing types and ratios. Both quench-cooling and incorporation into coamorphous systems improved the dissolution, solubility, and physical stability of individual APIs. Conclusions: Our findings provide deeper insight into the co-amorphization, dissolution, and stability characteristics of specific drug-drug coamorphous systems FEL-NAP and NTP-NAP, offering valuable guidance for developing new ddCAM coamorphous formulations containing some RT-unstable drugs.

Nanotherapies, valued for their high efficacy and low toxicity, frequently serve as antitumor treatments, but do not readily penetrate deep into tumor tissues and cells. Here we developed an improved tumor-penetrating peptide (TPP)-based drug delivery system. Briefly, the established TPP iNGR was modified to generate a linear NGR peptide capable of transporting nanotherapeutic drugs into tumors through a CendR pathway-dependent, neuropilin-1 receptor-mediated process. Although TPPs have been reported to reach intended tumor targets, they often fail to penetrate cell membranes to deliver tumoricidal drugs to intracellular targets. We addressed this issue by harnessing cell penetrating peptide technology to develop a liposome-based multibarrier-penetrating delivery system (mbPDS) with improved synergistic drug penetration into deep tumor tissues and cells. The system incorporated doxorubicin-loaded liposomes coated with nona-arginine (R9) CPP and cyclic iNGR (CRNGRGPDC) molecules, yielding Lip-mbPDS. Lip-mbPDS tumor-targeting, tumor cell/tissue-penetrating and antitumor capabilities were assessed using CD13-positive human fibrosarcoma-derived cell (HT1080)-based in vitro and in vivo tumor models. Lip-mbPDS evaluation included three-dimensional layer-by-layer confocal laser scanning microscopy, cell internalization/toxicity assays, three-dimensional tumor spheroid-based penetration assays and antitumor efficacy assays conducted in an animal model. Lip-mbPDS provided enhanced synergistic drug penetration of multiple biointerfaces for potentially deep tumor therapeutic outcomes. We co-modified R9 and iNGR on the liposome surface to obtain Lip-mbPDS. The penetration of Lip-mbPDS into multiple biointerfaces has enabled it to demonstrate good anti-tumor ability for CD13-positive HT1080 cells both in vivo and ex vivo.

Peptides and proteins, two important classes of biomacromolecules, play important roles in the biopharmaceuticals field. As compared with traditional drugs based on small molecules, peptide- and protein-based drugs offer several advantages, although most cannot traverse the cell membrane, a natural barrier that prevents biomacromolecules from directly entering cells. However, drug delivery via cell-penetrating peptides (CPPs) is increasingly replacing traditional approaches that mediate biomacromolecular cellular uptake, due to CPPs’ superior safety and efficiency as drug delivery vehicles. In this review, we describe the discovery of CPPs, recent developments in CPP design, and recent advances in CPP applications for enhanced cellular delivery of peptide- and protein-based drugs. First, we discuss the discovery of natural CPPs in snake, bee, and spider venom. Second, we describe several synthetic types of CPPs, such as cyclic CPPs, glycosylated CPPs, and D-form CPPs. Finally, we summarize and discuss cell membrane permeability characteristics and therapeutic applications of different CPPs when used as vehicles to deliver peptides and proteins to cells, as assessed using various preclinical disease models. Ultimately, this review provides an overview of recent advances in CPP development with relevance to applications related to the therapeutic delivery of biomacromolecular drugs to alleviate diverse diseases.

Despite the potential advantages of amorphism-induced supersaturation, the merit of new amorphization formation methods on the properties of the amorphous drug including the stability of the amorphous state, dissolution/solubility, supersaturation, and “spring-parachute” process is still poorly understood, particularly for certain amorphous supersaturating drug delivery systems (aSDDS). The present work aimed to explore the detailed merit of current attractive amorphization manufacturing methods (i. g., hot-melt extrusion (HME) technique) on the property improvement of aSDDS in form of amorphous solid dispersion microparticles by employing a model BCS II drug nitrendipine and a polyvinylpyrrolidone-based model polymer copovidone. Many aSDDS systems were developed by various methods, and their physicochemical properties were characterized by SEM, PXRD and DSC. HME-triggered amorphization induced superior supersaturation by the observation of the highest dissolution and solubility. HME induced the optimal supersaturation duration by the observed greatest extension of “spring-parachute” process (e. g., maximum AUCspring-parachute). HME technique is comparable with other techniques for the stabilization of amorphous state during storage. All aSDDS systems by HME and other methods showed improved long-term stability of the amorphous state in comparison to the pure amorphous drug. Fourier transformation infrared spectroscopy, Noyes-Whitney equation, nucleation theory and Gibbs free energy of transfer (ΔGto) were used to analyze the underlying mechanisms. Molecular mechanism studies indicated that HME caused a stronger crystallization inhibition effect in the aSDDS systems than other methods, but molecular interaction is not a dominant mechanism for property enhancement caused by HME. For the mechanism associated with the polymer itself (PVPVA64), it could inhibit the drug recrystallization, solubilize the drug spontaneously and cause the improved molecular interactions in all aSDDS systems. This study provided a deep insight into detailed advantage of HME-triggered supersaturation/amorphization and facilitated the applications of the technique both in the field of particuology and in pharmaceutical industry.

Aging has become an irreversible trend in the world, the health problems caused by aging cannot be ignored. The physiological functions of human body begin to decline with aging, the decline of gastrointestinal function caused by aging is an important problem that needs to be resolved. In this work, we evaluated the anti-aging effect of uridine in the senescent gastric epithelial cell model, and found that the aging level of gastric epithelial cell was significantly down-regulated by uridine treatment, uridine could obviously down-regulate the ratio of the SA-β-gal-positive senescent cells. Furthermore, aging-related marker molecules (such as p16 and p21) were also significantly down-regulated under uridine treatment. Additionally, the levels of inflammation and oxidative stress were also significantly reduced by uridine treatment. Next, our further studies the effect of aging on FGF activity on gastric epithelial cell, and found that FGF/FGFR-mediated signaling pathways were significantly down-regulated. However, uridine treatment can not only alleviate the senescence of gastric epithelial cell, but also can partially restore the sensitivity of FGF signaling. Taken together, the current work indicates that uridine shows a good anti-aging effect, which lays a solid foundation for the related research in this filed.

Amorphous solid-state dispersions that achieve supersaturation are often used to generate supersaturating drug delivery systems (SDDS). Relative to kinetic disordering approaches, thermodynamic disordering strategies utilizing currently popular self-micellizing amphiphilic polymers are better suited for industrial applications and can greatly improve supersaturating immediate-release of self-micellizing solid dispersion (SmSD) microcarrier delivery systems. However, the impact of thermodynamic disordering processes in different patterns (continuous and discontinuous) on the amorphization and supersaturating immediate-release of amorphous SmSD systems containing insoluble drugs is not clear. Here, we employed hot melt extrusion (HME) as a continuous thermodynamic disordering process, and two methods including solvent evaporation (SE) and microwave-quench cooling (MC) as discontinuous processes, to fabricate amorphous SmSD systems containing self-micellizing polymer Soluplus ® for the supersaturating delivery of a water-insoluble BCS II drug felodipine. Characterization of these amorphous SmSD systems was accomplished via scanning electron microscopy (SEM), powder X-ray diffraction (PXRD), and differential scanning calorimetry (DSC). HME-based continuous thermodynamic disordering process triggered a marked extension of supersaturating immediate-release by elevating solubility, enhancing dissolution and moving up “spring-parachute” processes as compared to discontinuous disordering processes. Molecular mechanisms contributing to internal stability of SmSD systems were explored through analyses of molecular interactions, crystallization inhibition effects and Gibbs free energy of transfer (ΔGtr°) values. This work indicates that HME-based continuous thermodynamic disordering process can be used in the development of superior amorphous SmSD microcarrier systems by triggering the generation of extended supersaturating immediate-release, and displays its further application potential in pharmaceutical industry.

The lack of effective rheumatoid arthritis (RA) therapies is a persistent challenge worldwide, prompting researchers to urgently evaluate traditional Chinese medicines (TCMs) as potential clinical RA treatments. The present investigation was conducted to evaluate the therapeutic effects and potential molecular mechanisms of the active components isolated from TCM Rhodiola sachalinensis Borissova from Baekdu Mountain (RsBBM) using an experimental adjuvant arthritis model induced by injection of rats with Freund's complete adjuvant. After induction of the adjuvant arthritis rat model, the extract-treated and untreated groups of arthritic rats were evaluated for RsBBM therapeutic effects based on comparisons of ankle circumferences and ELISA-determined blood serum inflammatory factor levels (TNF-α, IL-1β, and PGE2). In addition, the joint health of rats was evaluated via microscopic examination of hematoxylin-eosin-stained synovial tissues. Furthermore, to explore whether NF-κB and RANK/RANKL/OPG signaling pathways participated in observed therapeutic effects from a molecular mechanistic viewpoint, mRNA and protein levels related to the expression of nuclear factor kappa-B (NF-κB), osteoprotegerin (OPG), and receptor activator of nuclear factor kappa-Β ligand (RANKL) were analyzed via quantitative RT-PCR and Western blot analysis, respectively. Treatment of arthritic rats with the extract of RsBBM was shown to reduce ankle swelling, reduce blood serum levels of inflammatory factors, and alleviate arthritis-associated synovial inflammation and joint damage. Moreover, an RsBBM 50% ethanol extract treatment inhibited bone destruction by up-regulating OPG-related mRNA and protein expression and down-regulating RANKL-related mRNA and protein expression, while also reducing inflammation by the down-regulating of the NF-κB pathway activity. The results clearly demonstrated that the extract of RsBBM alleviated adjuvant arthritis-associated joint damage by altering activities of inflammation-associated NF-κB and the RANK/RANKL/OPG signaling pathways. Due to its beneficial effects for alleviating adjuvant arthritis, this RsBBM 50% ethanol extract should be further evaluated as a promising new therapeutic TCM treatment for RA.

The short-term immediate release of supersaturated drug-delivery systems (SDDSs) presents an interesting process that can be tailored to multi-stage release events including initial release after dosing and dissolution, evolved release over longer dissolution periods for biological absorption, and terminal release following the end of immediate release. However, although comprehensive analysis of these critical release behaviors is often ignored yet essential for understanding the supersaturable immediate-release events for supersaturable solid formations when employing new techniques or polymers matched to a particular API. Hot-melt extrusion (HME) has become a popular continuous thermodynamic disordering technique for amorphization. The self-micellizing polymer Soluplus® is reported to be a potential amorphous and amphiphilic graft copolymer frequently used in many nano/micro supersaturable formulations. Our current work aims to develop hypotensive supersaturating solid dispersion systems (faSDDSHME) containing the BCS II drug, felodipine, when coordinately employing the HME technique and self-micellizing Soluplus®, and to characterize their amorphization as well as immediate release. Other discontinuous techniques were used to prepare control groups (faSDDSSE and faSDDSQC). Tailored initial/evolved/terminal three-stage supersaturable immediate-release behaviors were identified and possible mechanisms controlling the release were explored. HME produced the highest initial release in related faSDDSHME. During the evolved-release period, highly extended "spring-parachute" process was found in HME-induced amorphization owing to its superior supersaturation duration. Due to the enhanced crystallization inhibition effect, faSDDSHME displayed the strongest terminal release as measured by solubility. For release mechanisms associated with HME, molecular interaction is not the likely dominant mechanism responsible for the improved properties induced by faSDDSHME. For release mechanisms involved with the polymer Soluplus® itself, they were found to inhibit drug recrystallization, spontaneously solubilize the drug and lead to improved molecular interactions in all SDDS systems, which were the factors responsible for the improved release. These mechanisms play an important role for the generation of an extended multi-stage immediate release produced via HME or self-micellizing polymer. This study provides a deeper understanding on amorphization and superior multi-stage supersaturable immediate-release behaviors for a particular hypotensive supersaturated delivery system combined with an HME-based continuous manufacturing technique and self-micellizing polymer strategy.

Hot melt extrusion (HME), a continuous manufacturing process for generating supersaturating amorphous self-micellizing solid dispersion systems (saSMSDs), holds promise for achieving amorphization of many pharmaceutical formulations. For saSMSDs generation, HME-triggered continuous processes offer advantages over traditional non-continuous processes such as fusion/quench cooling (FQC) and co-precipitation (CP). Here we employed HME, FQC, and CP to generate saSMSDs containing the water-insoluble BCS II drug nitrendipine (NIT) and self-micellizing polymer Soluplus®. Scanning electron microscopy, powder X-ray diffraction, and differential scanning calorimetry results revealed that saSMSDs formed when NIT–Soluplus® mixtures were subjected to the abovementioned amorphization methods. All saSMSDs outperformed crystalline NIT preparations and physical mixtures in achieving extended supersaturable immediate release states with superior solubility, “spring-parachute” process characteristics, and dissolution behaviors. Notably, Fourier transform-infrared spectroscopic results obtained for saSMSDs detected hydrogen bonding interactions between the drug and the carrier. Ultimately, our results revealed the advantages of HME-triggered amorphization as a continuous process for significantly improving drug dissolution, increasing solubility, and maintaining supersaturation as compared to traditional amorphization-based techniques.

Purpose: We aimed to investigate whether dihydromyricetin (DHM) could alleviate acetaminophen (APAP)-induced liver damage in mice, and to verify whether the process is associated with the PI3K/AKT signaling pathway. Methods: The contents of DHM in serum and related physiological indicators in blood and liver tissue were measured, respectively. We used haematoxylin and eosin (H&E), TUNEL, Hoechst 33,258, immunofluorescence assay and western blot methods to comprehensively assess the protective mechanism and therapeutic effect of DHM on liver damage induced by APAP (250 mg/kg) in mice. Results: APAP (250 mg/kg) could increase the expression of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-α (TNF-α), and interleukin 1β (IL-1β) and cause 4-hydroxy-2-nonenal (4-HNE) and Cytochrome P450 2E1 (CYP2E1) overexpression and stress response in the PI3K/AKT pathway. DHM was also detected in the serum of mice about five minutes after administration. DHM pretreatment could reverse GSH depletion and CYP2E1 overexpression, reduce the expression of ALT, AST, malondialdehyde, 4-HNE, TNF-α, and IL-1β, meanwhile it could reverse the abnormal expression of PI3K/AKT signaling pathway-related proteins which were induced by APAP. DHM pretreatment significantly reduced APAP-induced liver tissue apoptosis, necrosis, and inflammatory infiltration. Conclusion: DHM had a hepatoprotective effect on hepatotoxicity induced by APAP, which was shown by inhibiting oxidative stress and inflammatory responses, and reducing hepatocyte apoptosis by activating the PI3K/AKT signaling pathway.

目的 建立液质联用检测方法分离鉴定藜芦主要化学成分,并比较各成分与不同比例南沙参配伍前后经Caco-2单层细胞模型吸收转运的变化情况,从吸收转运的角度分析南沙参对藜芦的"增毒"机制.利用Elisa法测定南沙参与不同比例藜芦配伍前后免疫调节作用的变化情况,从药效作用的角度分析藜芦对南沙参的"减效"机制.方法 液质条件为乙腈(A)和0.1％甲酸水溶液(v)作流动相,梯度洗脱,Acquity UPLC BEH C18(1.7 μm,2.1 mm×50 mm)色谱柱,柱温设置35℃;3200 QTRAPTM串联四级杆-线性离子阱质谱仪,采用电喷雾电离源,电喷雾电压为5.0 kV,透镜管电压为90 V,金属毛细管电压为40V、温度为250℃.结果 利用液质联用方法分离鉴定出8种生物碱成分,发现在一定配伍比例下南沙参可使藜芦中一些生物碱溶出和生物利用度提高,具有"增毒"作用.而藜芦的加入,使南沙参的免疫调剂作用降低,并随着藜芦用量的增加其药效作用降低明显,具有"减效"的作用.结论 该研究结果从化学成分含量变化、吸收转运和药效学改变的角度再次考察了中药"十八反"中南沙参"叛"藜芦的配伍禁忌原则,为深入研究中药十八反中藜芦-南沙参药对提供理论基础.

目的 以长白山地区刺五加根为材料,采用响应面分析法优化刺五加总苷超声波提取工艺.方法 以提取工艺中料液比、乙醇体积分数和超声波处理时间为考察参数,以香草醛-高氯酸显色法测定的提取率为评价指标,利用Box-Behnken响应面法分析超声波提取刺五加总苷的最佳条件.结果 超声波提取刺五加总苷的最佳工艺参数为液料比1:34 g/mL,乙醇体积分数72.06％,超声波处理时间63.40 min.结论 响应面法优化刺五加总苷的超声波提取工艺合理可行,有效提高了提取率,为刺五加总苷的深入研究奠定基础.

对北五味子中总木脂素类成分进行纯化并研究其抗氧化和抗炎活性.利用大孔吸附树脂纯化五味子总木脂素,采用总抗氧化活性法、超氧阴离子自由基清除率法测定总木脂素类成分的抗氧化活性,应用酶联免疫吸附测试(enzymelinkedimmunosorbentassay,ELISA)和一氧化氮(NO)试剂盒考察五味子总木脂素类成分对脂多糖(lipopolysaccharide,LPS)诱导的小鼠巨噬细胞分泌肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(Interleukinin-10,IL-10)、NO的影响.抗氧化活性研究方面,与同等浓度的VC对照品相比,北五味子总木脂素类成分的总抗氧化能力是维生素C(VC)的1/7左右,清除超氧阴离子自由基的强度是VC的1/6左右,说明其具有良好的抗氧化活性.抗炎活性研究方面,北五味子总木脂素类成分可显著促进小鼠巨噬细胞分泌IL-10,抑制TNF-α、NO,说明其具有良好的抗炎活性.北五味子总木脂素类成分具有良好的抗氧化能力和抗炎活性,为进一步研究及开发北五味子提供理论支持.

Objective To observe the anti-enterovirus type 71(EV71)effects of Houttuynia cordata Thunb water ex-tract and its main ingredients,including chlorogenic acid,rutin,spasinin,and quercetin in vitro.Methods The RD cells (derived from human rhabdomyosarcoma cells)were divided into the Houttuynia cordata group,hyperoside group,querce-tin group,rutin group,chlorogenic acid group,and control group.Cells in the Houttuyniae cordata group were given water extracts of Houttuynia cordata at the concentrations of 5 000,2 500,1 250,625,and 313 mg/L;cells in the quercetin and rutin groups were added with quercetin and rutin at the concentrations of 10.00,5.00,2.50,1.25, and 0.63 mg/mL;cells in the hyperoside and chlorogenic acid groups were given hyperoside and chlorogenic acid,respectively,with the con-centrations of 4.00,2.00,1.00,0.50,and 0.25 mg/mL;cells in the control group were added with the cell culture flu-id.We observed the cytopathic effect(CPE)and calculated the inhibition rate of virus replication,half maximal inhibitory concentration(IC50),inhibition rate of virus growth,inhibition rate of virus adsorption, virus inactivation rate,and virus penetration rate.Results The inhibition rate of virus replication in the Houttuynia cordata group and quercetin group was higher than that in the other four groups, and the inhibition rate increased with the increasing concentrations(all P<0.05).The inhibition rate of virus growth in the hyperin and rutin groups was higher than that in the other four groups(all P<0.05).The inhibition rate of virus adsorption in the hyperin and Houttuynia cordata group was higher than that in the other four groups(all P<0.05).The water extract and its main components of Houttuynia cordata had no effect in inacti-vating the virus.No inhibitory effect on virus penetration was observed in each group at 15 and 30 min.At 60 min,the in-hibition rate of virus penetration was higher in houttuyniae,quercetin,rutin and chlorogenic acid groups than in the hypero-side group and control group(all P<0.05).The inhibition rate of virus penetration of the quercetin group and rutin group decreased with the decreasing drug concentrations, but increased in the chlorogenic acid group.Conclusions The water extract of Houttuynia cordata Thunb and its main components all have the effect of anti-EV71 in vitro,but had no effect in inactivating the virus.Quercetin has the effect of inhibiting virus replication.Hyperin and rutin block the growth of the vi-rus,hyperoside can inhibit virus adsorption,and quercetin,rutin,chlorogenic acid can inhibit the virus penetration.

[目的]以胰岛素信号通路中负调节蛋白PTP1B为靶点,筛选降糖药用真菌.[方法]比较了蛹虫草、蝙蝠蛾拟青霉、桑黄、灰树花4种药用真菌粗提物对PTP1B的抑制活性,并通过糖尿病小鼠模型进行验证.[结果]蝙蝠蛾拟青霉对PTP1B的抑制率最高,达到(78±2.81)％,其抑制类型为竞争性抑制.体内实验显示,蝙蝠蛾拟青霉降血糖效果最佳,小鼠空腹血糖降低了71.2％,与模型组有显著差异(p＜0.01),和二甲双胍组相近.蛹虫草组、灰树花组和桑黄组血糖分别降低了62.7％、57.8％和48.3％.此外,各个实验组小鼠的甘油三酯、胆固醇、糖化血红蛋白和胰岛素水平与模型组也存在显著差异.[结论] PTP1B作为糖尿病的新靶标,在筛选降糖药用真菌的过程中也同样适用,而且蝙蝠蛾拟青霉显示了良好的降血糖活性.