In Vitro and In Vivo Evaluation of Core–Shell Mesoporous Silica as a Promising Water-Insoluble Drug Delivery System: Improving the Dissolution Rate and Bioavailability of Celecoxib With Needle-Like Crystallinity

The objective of our study was to prepare mesoporous silica nanoparticles with a core–shell structure (CSMSNs) and improve the dissolution and bioavailability of celecoxib (Cxb), a water-insoluble drug, by changing its needle-like crystal form. CSMSNs are prepared by a core-shell segmentation self-assembly method. The S_BET and V_t of CSMSNs were 890.65 m²/g and 1.23 cm³/g, respectively. Cxb was incorporated into CSMSNs by the solvent evaporation method. The gastrointestinal irritancy of the CSMSNs was evaluated by a gastric mucosa irritation test. In vitro dissolution and in vivo pharmacokinetic tests were carried out to study the improvement in the dissolution behavior and oral bioavailability of Cxb. In conclusion, gastric mucosa irritation study indicated the good biocompatibility of CSMSNs. The cumulative dissolution of CSMSNs-Cxb is 86.2% within 60 min in SIF solution, which may be ascribed to the crystal form change caused by control of the nanochannel for CSMSNs. Moreover, CSMSNs could enhance the 9.9-fold AUC of Cxb. The cumulative dissolution and bioavailability of Cxb were both significantly enhanced by CSMSNs. CSMSNs with a core–shell structure are suitable as a carrier for a poorly water-soluble drug (Cxb).     

Celecoxib is a substrate of CYP2D6: Impact on celecoxib metabolism in individuals with CYP2C9*3 variants

Celecoxib was characterized as a substrate of human cytochrome P450 (CYP) 2D6 in vitro. In recombinant CYP2D6, celecoxib hydroxylation showed atypical substrate inhibition kinetics with apparent K_m, K_i, and V_max of 67.2 μM, 12.6 μM, and 1.33 μM/min, respectively. In human liver microsomes (HLMs), a concentration-dependent inhibition of celecoxib hydroxylation by quinidine was observed after CYP2C9 and CYP3A4 were inhibited. In individual HLMs with variable CYP2D6 activities, a significant correlation was observed between celecoxib hydroxylation and CYP2D6-selective dextromethorphan O-demethylation when CYP2C9 and CYP3A4 activities were suppressed (r = 0.97, P < 0.0001). Molecular modeling showed two predominant docking modes of celecoxib with CYP2D6, resulting in either a substrate or an inhibitor. A second allosteric binding antechamber, which stabilized the inhibition mode, was revealed. Modeling results were consistent with the observed substrate inhibition kinetics. Using HLMs from individual donors, the relative contribution of CYP2D6 to celecoxib metabolism was found to be highly variable and dependent on CYP2C9 genotypes, ranging from no contribution in extensive metabolizers with CYP2C9*1*1 genotype to approximately 30% in slow metabolizers with allelic variants CYP2C9*1*3 and CYP2C9*3*3. These results demonstrate that celecoxib may become a potential victim of CYP2D6-associated drug-drug interactions, particularly in individuals with reduced CYP2C9 activity.     

关节腔注射塞来昔布微球对大鼠类风湿性关节炎的治疗作用

考察关节腔注射塞来昔布（CEL）聚乳酸-羟基乙酸共聚物（PLGA）微球对大鼠类风湿性关节炎的治疗作用。以PLGA为载体,采用O/W乳化溶剂挥发法制备塞来昔布微球（CEL-MS）。采用完全弗氏佐剂诱导佐剂型关节炎（AA）大鼠模型,以大鼠足跖肿胀度、关节炎指数、脾脏指数和关节滑膜组织病理学检查作为治疗效果的指标,考察关节腔注射CEL-MS对大鼠类风湿性关节炎的治疗作用。结果显示,微球的形态圆整、表面光滑,平均粒径为（2.1 ± 0.3）μm、载药量为（20.8 ± 0.6）%。体内药效实验结果显示,与灌胃给予塞来昔布混悬液和关节腔注射塞来昔布混悬液相比,在关节炎大鼠关节腔中注射CEL-MS可显著降低关节肿胀和关节炎指数,有效抑制滑膜炎症。以上结果表明,关节腔注射CEL-MS对大鼠类风湿性关节炎具有良好的治疗作用。     

塞来昔布对离体人膝骨关节炎细胞凋亡及EGFR/MAPK信号通路的影响

目的 探究塞来昔布对离体人膝骨关节炎细胞凋亡及表皮生长因子受体/丝裂原激活蛋白激酶（EGFR/MAPK）通路的影响。方法 体外分离及培养膝关节软骨细胞,并以甲苯胺蓝染色鉴定软骨细胞。将细胞分为空白对照组（人正常膝骨关节软骨细胞不做特殊处理）、模型组（人膝骨关节炎软骨细胞不做特殊处理）及塞来昔布低、高剂量组（人膝骨关节炎软骨细胞中分别加入含终浓度为50 μmol/L和200 μmol/L塞来昔布溶液的培养液）。采用CCK-8法、AnnexinV/PITC双染法分别检测各组细胞增殖、凋亡能力,Western blotting检测细胞增殖、凋亡及EGFR/MAPK通路蛋白的表达。结果 甲苯胺蓝染色结果显示,人正常膝骨关节软骨细胞、人膝骨关节炎软骨细胞内见蓝紫色异染颗粒,细胞核呈深蓝色,细胞质呈浅蓝色;与人膝骨关节炎软骨细胞形态比较,人正常膝骨关节软骨细胞体积较大,形态较规则,蓝紫色异染颗粒较多。与空白对照组比较,模型组,塞来昔布低、高剂量组细胞存活率降低（P <0.05）,凋亡率升高（P <0.05）;与模型组比较,塞来昔布低、高剂量组细胞存活率升高（P <0.05）,凋亡率降低（P <0.05）;与塞来昔布低剂量组比较,塞来昔布高剂量组细胞存活率升高（P <0.05）,凋亡率降低（P <0.05）。与空白对照组比较,模型组,塞来昔布低、高剂量组EGFR、MMP-9、Caspase-3蛋白相对表达量升高（P <0.05）,p-P38 MAPK/P38 MAPK蛋白相对表达量降低（P <0.05）;与模型组相比,塞来昔布低、高剂量组EGFR、MMP-9、Caspase-3蛋白相对表达量降低（P <0.05）,p-P38 MAPK/P38 MAPK蛋白相对表达量升高（P <0.05）;与塞来昔布低剂量组比较,塞来昔布高剂量组EGFR、MMP-9、Caspase-3蛋白相对表达量降低（P <0.05）,p-P38 MAPK/P38 MAPK蛋白相对表达量升高（P <0.05）。结论 塞来昔布能促进离体人膝骨关节炎细胞增殖,抑制细胞凋亡。其作用可能是通过抑制EGFR表达,促进P38 MAPK通路活化实现的。     

Reduction of arthrofibrosis utilizing a collagen membrane drug-eluting scaffold with celecoxib and subcutaneous injections with ketotifen

The dense formation of abnormal scar tissue after total knee arthroplasty results in arthrofibrosis, an unfortunate sequela of inflammation. The purpose of this study was to use a validated rabbit model to assess the effects on surgically-induced knee joint contractures of two combined pharmacological interventions: celecoxib (CXB) loaded on an implanted collagen membrane, and subcutaneously (SQ) injected ketotifen. Thirty rabbits were randomly divided into five groups. The first group received no intervention after the index surgery. The remaining four groups underwent intra-articular implantation of collagen membranes loaded with or without CXB at the time of the index surgery; two of which were also treated with SQ ketotifen. Biomechanical joint contracture data were collected at 8, 10, 16, and 24 weeks. At the time of necropsy (24 weeks), posterior capsule tissue was collected for messenger RNA and histopathologic analyses. At 24 weeks, there was a statistically significant increase in passive extension among rabbits in all groups treated with CXB and/or ketotifen compared to those in the contracture control group. There was a statistically significant decrease in COL3A1, COL6A1, and ACTA2 gene expression in the treatment groups compared to the contracture control group (P < .001). Histopathologic data also demonstrated a trend towards decreased fibrous tissue density in the CXB membrane group compared to the vehicle membrane group. The present data suggest that intra-articular placement of a treated collagen membrane blunts the severity of contracture development in a rabbit model of arthrofibrosis, and that ketotifen and CXB may independently contribute to the prevention of arthrofibrosis. Statement of clinical significance: Current literature has demonstrated that arthrofibrosis may affect up to 5% of primary total knee arthroplasty patients. For that reason, novel pharmacologic prophylaxis and treatment modalities are critical to mitigating reoperations and revisions while improving the quality of life for patients with this debilitating condition.     

塞来昔布对压力性尿失禁模型大鼠的效果及作用机制

目的研究塞来昔布对压力性尿失禁模型大鼠的干预效果及作用机制。方法建立压力性尿失禁大鼠模型后分为模型组和塞来昔布组各18只,正常组10只。塞来昔布组大鼠采用塞来昔布进行干预,模型组和正常组大鼠采用等体积的无菌蒸馏水干预,设置喷嚏试验、检测尿流动力学指标,然后测定漏尿点压力(LPP)、腹部漏尿点压力(ALPP),观察3组大鼠病理形态学特征,Western blot检测转化生长因子-β1(TGF-β1)、结缔组织生长因子(CTGF)通路蛋白表达,酶联免疫吸附试验法检测环氧化酶2(COX-2)、脂肪氧化酶(LOX)、基质金属蛋白酶-2(MMP-2)蛋白水平。结果喷嚏实验阳性率模型组66.67%,显著大于塞来昔布组的27.78%,差异具有显著性统计学意义(P<0.05);LPP、ALPP、TGF-β1、CTGF水平及LOX蛋白水平均为模型组<塞来昔布组<正常组(P均<0.05),COX-2、MMP-2蛋白水平均为模型组>塞来昔布组>正常组(P均<0.05)。结论塞来昔布对压力性尿失禁模型大鼠的治疗效果显著,通过调控TGF-β1/CTGF通路蛋白,可以改善大鼠COX-2、LOX、MMP-2水平,对于治疗和预防压力性尿失禁的研究提供了方向。     

塞来昔布联合吉非替尼对HCC827细胞凋亡影响机制探讨

目的探讨Cox-2抑制剂塞来昔布联合吉非替尼对肺癌EGFR 19号外显子E746-A750缺失细胞株HCC827细胞凋亡的影响及其可能机制。方法 RPMI1640培养HCC827细胞,分为正常对照组、塞来昔布组、吉非替尼组、塞来昔布加吉非替尼组。塞来昔布与吉非替尼药物浓度均为5、10、20、40、80μmol/L。药物干预细胞48h后,MTT法检测细胞增殖,流式细胞术检测细胞凋亡,蛋白质印迹法检测细胞中Cox-2和p-EGFR蛋白表达。结果 MTT结果显示,塞来昔布和吉非替尼单独用药时HCC827细胞的增殖受到明显的抑制,且随着药物浓度的增加,抑制作用逐步增强,药物剂量与细胞增殖抑制率呈正相关;塞来昔布与吉非替尼联合用药时对HCC827细胞呈现出更强的抑制作用,与单独用药相比差异有统计学意义,P<0.001;对照组细胞凋亡率为(0.26±0.09)%,塞来昔布组为(4.86±0.37)%,吉非替尼组为(8.53±0.78)%,塞来昔布+吉非替尼组为(23.28±1.63)%,组间比较差异有统计学意义,F=111.291,P<0.001,且两药联合时具有交互效应,F=90.440,P<0.001;蛋白质印记法结果显示,用药组较对照组Cox-2(F=185.351,P<0.001)和p-EGFR(F=61.328,P<0.001)蛋白水平均有不同程度的下调,其中两药联合组下调最为明显。结论塞来昔布与吉非替尼具有良好的协同作用,其机制可能与诱导凋亡、抑制EGFR的活化和下调Cox-2蛋白的表达有关,在治疗非小细胞肺癌中联合用药可能会具有较大的应用潜力。     

EFFECTS OF p53 GENE THERAPY COMBINED WITH CYCLOOXYGENASE-2 INHIBITOR ON CYCLOOXYGENASE-2 GENE EXPRESSION AND GROWTH INHIBITION OF HUMAN LUNG CANCER CELLS

1 INTRODUCTIONCOXis the key enzyme involved in the synthesis of prostanoids,a collective termfor the PGs andthromboxanes.Of the two major isoforms of the COXenzyme,COX-1 is ubiquitous and constitutively ex-pressedin virtually all normal tissues.In contras…     

Effects of Combined Application of Muscle Relaxants and Celecoxib Administration After Total Knee Arthroplasty (TKA) on Early Recovery: A Randomized,Double-Blind,Controlled Study

The purpose of this study was to evaluate the effectiveness of application of muscle relaxants and celecoxib in early recovery after total knee arthroplasty (TKA). One hundred and fifty patients were randomized 1:1:1 to receive either both of muscle relaxants and celecoxib or muscle relaxants alone or placebo for 2 weeks (50 patients in each group). VAS pain scores as primary efficacy, active range of motion, morphine consumption, blood loss, and postoperative complications including postoperative nausea and vomiting (PONV), extremities myasthenia and deep vein thrombosis (DVT) were determined postoperatively. Group A improved better with reduced VAS pain scores compared with another two groups. These results demonstrated that application of muscle relaxants and celecoxib into patients undergoing TKA for 2 weeks postoperative consequently improved their convalescence.