Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease with limited treatment options and high mortality rate. Pyroptosis mediated by gasdermin D (GSDMD) has been linked to disease advancement by releasing pro-fibrotic mediators, whereas the serum amyloid P component (SAP) shows anti-fibrotic properties. In this study, we designed a pulmonary mucus-penetrating lipid nanoparticle delivery system, known as DAS-lipid nanoparticle (DAS-LNP), to enable the simultaneous administration of GSDMD-targeting small interfering RNA (siRNA) and SAP-expressing plasmid DNA (si/pD-DAS-LNP). The formulation showed enhanced mucus penetration (approximately 2.2-fold) and improved endo-lysosomal escape, leading to a 3.5-fold increase in lung accumulation of the nucleic acid drug compared to the free drug group, and a 1.5-fold increase relative to the standard MC3-LNP (MC-LNP) group at 24 h post-administration. Both in vitro and in vivo studies indicated a favorable safety profile for DAS-LNP. In a bleomycin-induced IPF mouse model, si/pD-DAS-LNP alleviated pulmonary inflammation, reduced pro-fibrotic factor levels, inhibited extracellular matrix deposition, improved lung function, and enhanced survival rates. The delivery efficiency enabled reduced dosage and administration frequency without compromising efficacy, thereby enhancing treatment compliance. These results suggest that concurrent inhibition of GSDMD and enhancement of SAP offer a complementary approach to modulate pyroptosis and the fibrotic microenvironment in IPF. The structure of DPPC/AS-modified LNP facilitates the effective pulmonary co-delivery of siRNA and pDNA, positioning si/pD-DAS-LNP as a promising translational strategy for IPF treatment.

This study aimed to evaluate and develop a novel long-acting injectable microcrystalline suspension of a proprietary prodrug, brexpiprazole laurate (BPZL), to improve the compliance of patients with schizophrenia. The BPZL microcrystalline suspension was prepared through a novel "progressive particle-size control" method, incorporating microfluidics, rapid freezing, and lyophilization. The formula was characterized by scanning electron microscope, X-ray diffraction analysis, differential scanning calorimetry, and dynamic light scattering analysis. Both in vitro and in vivo experiments were conducted to assess the performance of the suspension. The resulting suspensions exhibited consistent particle size distributions (D10: 1.23 +/- 0.19 & micro;m, D50: 2.62 +/- 0.56 & micro;m, D90: 4.88 +/- 1.28 & micro;m) and demonstrated excellent stability over 67 days of storage at 25 degrees C. In vivo pharmacokinetic studies in rats showed that there was no initial burst release after a single intramuscular administration, and the plasma concentration of the active metabolite (brexpiprazole) remained above the therapeutic threshold (the median effective concentration, EC50) for approximately 5 days. These results demonstrated that this technique effectively enhances the microcrystalline formulation's long-term stability and sustained-release performance while preventing sudden release effects. These findings offer theoretical support and practical guidance for the clinical application of BPZL microcrystals and the development of sustained-release formulations for other lipophilic drugs.

Current analgesics on the market exhibit a short duration of action and induce the production of inflammatory factors in tissues damaged by surgical procedures. Inflammatory factor production can create acidic environments, limiting drug delivery. In this study, we developed a novel injectable formulation comprising bupivacaine multivesicular liposomes of high osmotic pressure (H-MVL) and meloxicam nanocrystals (MLX) in a thermosensitive gel (H-MVL/MLX@GEL) adapted to the microenvironment for long-term postoperative analgesia. To achieve formulation stability, H-MVL were prepared by regulating the osmotic pressure of the gel system. Moreover, the inclusion of MLX serves to not only attenuate local inflammatory factors, regulating the acidic microenvironment, but also to prolong the duration of action of meloxicam (MEL). The increased absorption of bupivacaine (BUP) and the prolongation of the half-life of BUP release in H-MVL/MLX@GEL were demonstrated through pharmacokinetic experiments. Sciatic nerve block models and hot plate analgesia tests demonstrated that H-MVL/MLX@GEL effectively alleviated pain for at least five days. The immunohistochemical results showed that the addition of MLX reduced the production of the local inflammatory factors interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α), thereby improving the analgesic effect by regulating the local acidic environment and alleviating local irritation.

Sepsis is a life-threatening condition caused by a dysregulated response to infection. Currently, no effective therapeutic drugs have been approved. Gasdermin D (GSDMD) deficiency has been shown to alleviate sepsis, therefore, small interfering RNA (siRNA)-based agents targeting GSDMD may be an effective approach for treating sepsis. Moreover, the development of lipid nanoparticles (LNPs) and the innovation of microfluidic technology have promoted the clinical translation of siRNA-based therapeutics. Here we designed a novel continuous microfluidic platform (CA/SAR/C1), leveraging the benefits of circular asymmetric split-and- recombination (CA/SAR) approach, to prepare superior LNPs encapsulating siRNA specific to GSDMD (siGSDMD) for treating sepsis. The siGSDMD-LNPs, with exceptional characteristics, precise controllability and excellent stability, were produced continuously. In vitro, the siGSDMD-LNPs were efficiently taken up by bone marrow-derived macrophages (BMDMs), promoted lysosomal escape, silenced GSDMD and subsequently reduced inflammatory factors secretion. In vivo studies conducted in a lipopolysaccharide (LPS)-induced septic mice revealed that pulmonary administration of siGSDMD-LNPs alleviated organ damage and improved survival rates, which were achieved by silencing GSDMD and thereby blocking pyroptosis. Our findings support siGSDMD-LNPs as a promising therapeutic strategy for the management of sepsis.

Co-crystallization has received widespread attention, particularly in the modification of poorly water-soluble drugs. In this paper, two new complexes were obtained from heterocyclic co-formers, namely relugolix-pyrazine and relugolix-2-picolinic acid. Relugolix-pyrazine was identified as a cocrystal, while relugolix-2-picolinic acid was identified as a salt based on single crystal analysis. The introduction of co-former induced a conformational alteration in relugolix to some degree and stabilized the structure through weak intermolecular forces. Relugolix-pyrazine exhibited improved solubility, stability and drug-target interactions, effectively addressing the limitations of the parent molecule regarding its physicochemical properties. Additionally, we developed computational simulations to predict the solubility and proposed a method to predict unknown substances which include cocrystals and salts. The simulations aimed to elucidate the dissolution behavior of these substances, and the results were consistent with the experimental data, thereby demonstrating that the formation of cocrystals/salts is thermodynamically favorable for their dissolution.

Microfluidic technology facilitates precise control over fluid mixing and interactions between the components, including self-assembly and precipitation. It offers new options for accurately manufacturing particles and holds significant potential in advancing micro/nanoparticle drug delivery systems (DDSs). Various microchannel/microfluidic chips have been explored to construct micro/nanoparticle DDSs. The precise manipulation of particle size, morphology, structure, stiffness, surface characteristics, and elasticity through microfluidic technology relies on specific microchannel geometrical designs and the application of exogenous energy, adhering to the principles of fluid motion. Consequently, this enables reproducible control over critical quality attributes (CQAs), such as particle size and distribution, encapsulation efficiency, drug loading, in vitro and in vivo drug delivery profiles, Zeta potential, and targeting capabilities, for micro/nanoparticle DDSs. In this review, we categorize microfluidic techniques and explore recent research developments in novel microchannel structures spanning the past 5 years (2018–2023) and their applications in micro/nanoparticle DDSs. Additionally, we elucidate the latest manipulation strategies of microfluidic techniques that impact foundational structures related to the CQAs of micro/nanoparticle DDSs. Furthermore, we offer insights into the industrial applications and challenges microfluidic techniques face in the context of novel micro/nanoparticle DDSs.

Cerebral ischemia-reperfusion injury（CI/RI） remains the main cause of disability and death in stroke patients due to lack of effective therapeutic strategies. One of the main issues related to CI/RI treatment is the presence of the blood-brain barrier（BBB）, which affects the intracerebral delivery of drugs. Ginkgolide B（GB）, a major bioactive component in commercially available products of Ginkgo biloba, has been shown significance in CI/RI treatment by regulating inflammatory pathways, oxidative damage, and metabolic disturbance, and seems to be a candidate for stroke recovery. However, limited by its poor hydrophilicity and lipophilicity, the development of GB preparations with good solubility, stability, and the ability to cross the BBB remains a challenge. Herein, we propose a combinatorial strategy by conjugating GB with highly lipophilic docosahexaenoic acid（DHA） to obtain a covalent complex GB-DHA, which can not only enhance the pharmacological effect of GB, but can also be encapsulated in liposomes stably. The amount of finally constructed Lipo@GB-DHA targeting to ischemic hemisphere was validated 2.2 times that of free solution in middle cerebral artery occlusion（MCAO） rats. Compared to the marketed ginkgolide injection, Lipo@GB-DHA significantly reduced infarct volume with better neurobehavioral recovery in MCAO rats after being intravenously administered both at 2 h and 6 h post-reperfusion. Low levels of reactive oxygen species（ROS） and high neuron survival in vitro was maintained via Lipo@GB-DHA treatment, while microglia in the ischemic brain were polarized from the pro-inflammatory M1 phenotype to the tissue-repairing M2 phenotype, which modulate neuroinflammatory and angiogenesis. In addition, Lipo@GB-DHA inhibited neuronal apoptosis via regulating the apoptotic pathway and maintained homeostasis by activating the autophagy pathway. Thus, transforming GB into a lipophilic complex and loading it into liposomes provides a promising nanomedicine strategy with excellent CI/RI therapeutic efficacy and industrialization prospects.

金合欢素(1)是一种选择性高、体内外抗房颤活性良好的黄酮化合物,但水溶性差和半衰期短的缺陷严重限制了其成药.为改善其水溶性、增强口服吸收率,该研究以肠道上的钠-葡萄糖转运蛋白1(SGLT1)为靶点,设计并合成了 2个1糖苷前药.Caco-2细胞转运试验结果表明,1糖苷前药的渗透率均比1有显著提高,其转运由SGLT1介导,且受到多药耐药蛋白的外排作用.大鼠药动学研究结果显示,1的单糖前药和二糖前药的cmax值分别比1提高5和30倍,AUC相比1分别提升10和100倍,半衰期均延长2h左右.以上研究结果表明,使用糖基修饰1可有效增强其口服吸收效率.

The functional properties of xanthan gum (XG) in pharmaceutical preparations depend on its rheological properties, which inevitably rely on its molecular structure. Hence, this work investigated the relationship between the molecular structure of XG and its rheological properties and functional characteristics, and revealed the structural factors influencing the XG functionalities in oral suspensions and matrix tablets. Primarily, the molecular structures of four commercial XG products were characterized by infrared spectroscopy, differential scanning calorimetry and measuring the monosaccharide composition, average molecular weight, and pyruvate and acetyl contents. Furthermore, the flow behavior and viscoelasticity of XG solutions, the viscoelasticity of XG hydrogels, and XG combinations (XGC, aqueous solution containing XG, liquid glucose, and glycerin) were investigated. Finally, the dissolution time of XGC and the swelling and erosion properties of the XG matrix were studied to evaluate XG functionality in oral suspensions and matrix tablets, respectively. Results showed that the polydispersity of molecular weight and the pyruvate content affected the functionality and performance of XG in suspension and tablet forms. The higher polydispersity and pyruvate content of XG improved the hydrogel strength, which led to a longer dissolution time of XGC and a higher swelling extent of the XG matrix but a slower erosion rate.

Purpose:To enhance tumor penetration by formulation design and tumor microenvironment (TME) modulation, herein a novel reactive oxygen species (ROS)-responsive size/shape transformable lipid-polymer hybrid nanoparticle (LPN) has been fabricated for the co-delivery of an anticancer and collagen-inhibition drug.Methods:A ROS-responsive poly(D, L-lactide)-thioketal-polyethylene glycol (PLA-TK-PEG) co-polymer was synthesized. LPNs were then fabricated by encapsulation of losartan (LST)-loaded micelles as the core to support paclitaxel (PTX)-loaded liposomes. The PEG content in the lipid shell of LPNs was then adjusted to obtain the size-/shape-transformable LPNs (M/LST-Lip/PTX-PEG5%). The ROS-responsiveness was observed in vitro by transmission electron microscopy and the tumor-penetration of the LPNs was evaluated in 3D tumor spheroids by confocal laser scanning microscopy. Tumor-targeting, tumor-penetrating, and antitumor efficacies of the NPs in 4T1 tumor-bearing mice were determined by in vivo imaging.Results:ROS-responsive micellar core degradation and the transformation of spherical LPNs (120nm) to smaller 40 mm discoid nanoparticles (NP) were observed. The transformable LPNs exhibited enhanced capacity of penetration in contrast to the un-transformable preparations in three-dimensional (3D) tumor spheroids. Furthermore, synergetic penetrating enhancement was achieved by LST-loaded transformable LPNs in 4T1 and fibroblast cell mixed 3D tumor spheroids. The improved tumor penetration of LST-loaded transformable LPNs was observed in vivo, which could be due to their collagen inhibiting and size/shape transformable effect. Due to their enhanced penetrability, LST and PTX-loaded transformable LPNs demonstrated significant in vivo antitumor efficacy in comparison to other preparations.Conclusion:The results confirmed the efficacy of M/LST-Lip/PTX-PEG5% in tumor targeting, collagen inhibition in TME, and enhanced tumor penetration. This novel drug delivery system can therefore play a substantial role in improving the therapeutic efficacy of antitumor drugs combined with TME-improving agents.

Liver mitochondria isolated from fetal and newborn guinea pigs synthesized phosphoenolpyruvate at 4-6 nmol/min per mg protein with 2 mM malate, succinate, and alpha-ketoglutarate as substrates. These rates were 90-110% of that by adult liver mitochondria and were not substantially altered in the second half of gestation or within 24 h after birth. Both palmitoyl- and octanoylcarnitine were inhibitory to phosphoenolpyruvate synthesis in adult and fetal preparations, but free octanoate was inhibitory only in adult liver mitochondria.

Docetaxel (DTX) is a poorly soluble drug. The purpose of this study was to explore a DTX-loaded micelle delivery system using N-(all-trans-retinoyl)-L-cysteic acid methyl ester sodium salt (XMeNa) as the carrier materials. In this study, amphiphilic surfactant XMeNa was synthesized. Then, the blood biocompatibility and the value of critical micelle concentration (CMC) were assessed by a hemolysis test and pyrene-based fluorescent probe techniques, respectively. The XM-DTX micelles were prepared using the method of thin-film hydration, and characterized by dynamic light scattering and transmission electron microscopy (TEM). The entrapment efficiency (EE) and drug loading efficiency (DLE) were assessed by the ultrafiltration method. In vitro release and pharmacokinetic behaviors of XM-DTX micelles were performed in rats using Taxotere (a commercialized DTX injection) as a control. Our data confirmed the excellent blood biocompatibility of XMeNa as a carrier. XMeNa can self-assemble into micelles in aqueous media with a very low CMC (6.2 μg/mL). The average size and zeta potential of the XM-DTX micelles were 17.3 ± 0.2 nm, and −41.6 ± 0.3 mV, respectively. EE and DLE reached up to 95.3 ± 0.7% and 22.4 ± 0.2%, respectively, which may account for the high solubility of DTX in normal saline. The micelles were spherical in TEM with good dispersion and no aggregation and adhesion, and exhibited good stability after reconstitution over 8 hours. Results from in vitro release assay suggested a much slower release behavior of XM-DTX micelles in comparison to Taxotere. Additionally, XM-DTX micelles prolonged DTX retention in blood circulation, increased the area under the curve by 2.4-fold, and significantly decreased the clearance of the drug. Given above, the XM-DTX micelles could improve the solubility and the release of DTX. The amphiphilic surfactant XMeNa also exhibited great potential as a vehicle for exploring delivery of poorly water soluble drugs in the future.

Doxorubicin hydrochloride (DOX) is one of the widely used antineoplastic agents in treating various cancers, yet it is always associated with the occurrence of adverse reactions that limit its clinical use. Currently, encapsulating DOX in micelles may represent a promising strategy to reduce toxicity and side effects of the drug. This study aimed to explore a novel acitretin-based surfactant (ACMeNa) with good solid stability to encapsulate DOX to form micelles (ACM-DOX). In this work, ACM-DOX micelles were prepared by a microfluidic method free of organic solvents. The characteristics of ACM-DOX micelles were assessed, including morphology, particle size, stability, entrapment efficiency, and drug loading. An in vitro cytotoxicity experiment of the micelles on MDA-MB-231 (a human breast cancer cell line) was also performed. The micelle formation mechanism suggested that the insoluble ACMeNa/DOX complex was formed by electrostatic interaction, and subsequently encapsulated by self-assembly into micelles. The designed ACM-DOX micelles had an average particle size of 19.4 ± 0.2 nm and a zeta potential of −43.7 ± 2.4 mV, with entrapment efficiency and drug loading efficiency of 92.4 ± 0.5% and 33.4 ± 0.3%, respectively. The ACM-DOX micelles had worm-like structures under a Cryo-transmission electron microscope and exhibited good stability within 8 hours after reconstitution and 4- to 32-fold dilution of its reconstituted solution. ACM-DOX micelles released 80% of DOX within 24 hours in a medium of pH = 5.0, and its drug profile can be described by a first-order model. Moreover, ACM-DOX micelles showed cytotoxicity against MDA-MB-231 in a dose-dependent manner, and displayed a higher antitumor activity as compared with free DOX, with IC50 values of DOX and ACM-DOX micelles being 6.80 ± 0.50 and 4.64 ± 0.32 μg/mL, respectively. Given above, ACMeNa has great application potential as a DOX carrier for the treatment of cancers.

Abstract Parathyroid hormone (PTH)1–34 is an effective peptide drug for osteoporosis therapy. However, the half-life of PTH1–34 in vivo is short, leading to the need for frequent injections of this drug during its treatment. To prolong the half-life of PTH1–34, a novel PTH1–34 analog was generated based on fatty acid generation, and its synthesis process included recombinant protein expression, side-chain modification, and peptide decoration. The PTH1–34 variant was expressed in Escherichia coli, with a single Lys (position 27) retained as a modification site. The side chain, –AEEA-γGlu-C18 diacid, was synthesized using 2-chlorotrityl chloride resin as a solid support, and then was conjugated to the PTH1-34 variant to form PTH-Lys27-AGC. Reversed-phase chromatography confirmed a high final purity (>98%) of the target compound; in vitro bioactivity tests showed that PTH-1 receptor potency of PTH-Lys27-AGC was comparable to that of the native PTH1–34. A competitive human serum albumin binding test demonstrated a high albumin affinity of PTH-Lys27-AGC in comparison to PTH1–34. In summary, we developed a novel PTH1–34 analog, PTH-Lys27-AGC, which may be a long-acting agent for osteoporosis treatment in the future.

Hypoparathyroidism (HP) is a rare disease with clinical manifestations of hypocalcemia and hyperphosphatemia, resulting from deficient or absent parathyroid hormone (PTH) secretion. Conventional treatment for patients with HP involves extensive calcium and vitamin D supplementation. In 2015, PTH1-84 was approved by the United States Food and Drug Administration as an adjunct for HP patients who cannot be well-controlled on conventional treatment. However, PTH1-84 therapy requires a daily injection, leading to poor patient compliance. The purpose of this study was to develop a long-acting PTH1-34 analogue by increasing its affinity to albumin. Three PTH1-34 variants were generated by substituting two of the three lysine (Lys) residues with arginine, reserving a single Lys as the modification site in each sequence. A series of side chains, containing fatty acid, deoxycholic acid, or biotin groups, were synthesized to modify these PTH1-34 variants by using a solid-liquid phase synthesis approach. In vitro bioactivity and albumin affinity tests were used to screen these new PTH1-34 analogues. Finally, Lys27-AAPC was selected from 69 synthesized analogues as a candidate therapeutic compound because it retained potency and exhibited a high albumin-binding capacity. In pharmacodynamic experiments, Lys27-AAPC demonstrated enhanced and prolonged efficacy in serum calcium elevating relative to PTH1-84. Moreover, a lyophilized powder for injection containing Lys27-AAPC was developed for further testing and represented a potential long-acting HP treatment.

本研究采用基因重组大肠埃希菌发酵的方法,以可溶形式表达人甲状旁腺激素融合蛋白.菌体经高压破壁和加热沉淀后获得融合蛋白,经Ulp1酶切去除泛素标签后获得目的蛋白粗品,经阳离子交换色谱法和反相色谱法获得纯度为99.2％的目的蛋白.产品经相对分子质量测定、肽图分析和生物活性测定,证明自制甲状旁腺激素的结构和功能与理论值一致.本研究为工业化制备重组人甲状旁腺激素提供了参考.

本研究选择两阶段的释药方法,其中酸阶段pH 1.0、缓冲阶段pH 6.8,分别模拟胃和小肠的酸碱环境,考察了释放介质的离子强度和离子种类对自制艾司奥美拉唑镁肠溶微丸释药行为的影响.结果 表明,释放介质的离子强度和阴离子种类对肠溶微丸的释药行为具有显著影响.在0.091～0.262 mol/L的离子强度范围内,肠溶性聚合物Eudragit L30D-55和主药艾司奥美拉唑镁的溶解性质均对释放介质的离子强度较敏感,是造成肠溶微丸的释药行为受释放介质离子强度显著影响的原因.而阴离子种类(盐酸盐和磷酸盐)对Eudragit L30D-55的溶解行为有显著影响,对主药几乎无影响,因此肠溶微丸的释药行为受到显著影响而未包肠溶衣的载药微丸的释药行为几乎不受影响.试验中未见阳离子(钾离子和钠离子)对肠溶微丸的释药行为产生显著性影响.因此,在肠溶制剂体外评价时应将离子强度、离子种类等因素的影响考虑在内.

本研究建立了一种以大肠埃希菌作为宿主,表达重组猪促肾上腺皮质激素(pig adrenocorticotropic hormone,pACTH)的新方法.pACTH在N末端与经典猪瘟病毒N末端自切蛋白酶Npro突变体EDDIE融合,然后在大肠埃希菌胞浆中以包涵体形式表达,包涵体约占菌体湿重的36.2％.经过复性后的包涵体可以促使EDDIE自酶切,从而获得pACTH.通过优化复性条件,EDDIE-pACTH蛋白的自切率从25％提高至60％.复性液通过酸沉淀,能够去除杂蛋白,经反相色谱纯化后获得纯度为99.56％的pACTH.

Background: Vincristine is a potent therapeutic agent with well-defined activity against hematologic malignancies and solid tumors. It is a cell-cycle specific drug with concentration and exposure duration dependent activity. When used by liposomal delivery, it exhibits enhanced anti-tumor activity. However, vincristine liposome formulation in the clinic is supplied as a 3-vial-kit due to lacking sufficient stability. So it has to be prepared in situ prior to use through a multi-step process. Purpose: The purpose here is to develop a more stable and ready-to-use liposomal formulation for vincritstine in one vial. Patients and methods: A series of preparations were investigated based on sphingomyelin/cholesterol/PEG2000-DSPE lipid composition, with different drug/lipid (D/L) ratios (1/10, 1/5, 1/2), using an active sucrose octasulfate triethylamine salt gradient loading method. In this work, compared to generic vincristine sulfate liposome injection (GVM), the stability both in vivo and in vitro and efficacy in vivo of novel vincristine liposomes were investigated. Results: It was shown that the degradation of vincristine during 2-8°C storage was significantly decreased from 8.2% in 1 month (GVM) to 2.9% in 12 months (D/L ratio 1/5). The half-time for sphingomyelin/cholesterol/PEG2000-DSPE liposomes in vivo could be adjusted from 17.4 h (D/L ratio 1/10) to 22.7 h (D/L ratio 1/2) in rats, while the half-time for GVM was only 11.1 h. The increase in drug retention contributed to the lower in vivo toxicity. The antitumor efficacy was evaluated using a human melanoma tumor model and showed remarkable improvement compared to GVM. Conclusion: The study demonstrates that the new formulation with the drug/lipid ratio of 1/5 owns a higher encapsulation efficiency, better stability, lower toxicity and superior antitumor efficacy, which is screened out for further development.