1H和19F核磁共振定量法测定塞来昔布含量

目的 分别采用_¹H核磁共振定量法和_¹⁹F核磁共振定量法测定塞来昔布绝对含量,并将两种方法测定结果与质量平衡法测定结果进行比较。方法 氢核磁共振定量法以塞来昔布δ 6.92处质子峰作为定量峰,马来酸δ 6.33处为内标峰。在弛豫延迟时间10 s,采样时间4.01 s,扫描次数为32条件下采集混合物的氢谱。氟核磁共振定量法以4-溴-2-氟-乙酰苯胺为内标,在谱宽240 ppm,中心频率-94.26 ppm,样品扫描次数为16次条件下采集氟谱,计算两种核磁共振定量法的测试结果。结果 氢核磁共振定量法和氟核磁共振定量法测定结果相近,且与质量平衡法测定结果基本一致。结论 核磁共振定量法可以用来测定塞来昔布含量,具有准确、高效优势。     

HPLC法测定塞来昔布原料药中有关物质

目的建立测定塞来昔布原料药中有关物质的高效液相色谱(HPLC)法。方法建立HPLC法,采用Century SIL C30色谱柱(250mm×4.6mm,5μm),流动相:水(A)–乙腈(B)–甲醇(C),梯度洗脱;体积流量:1.0mL/min;检测波长:215 nm;柱温:25℃;进样量:20μL。结果塞来昔布与各杂质的分离度良好,且在一定浓度范围内线性关系良好。塞来昔布和杂质A、B、C、D、F的定量限分别为198.0、161.2、240.0、239.4、203.5、156.8 ng/mL,检测限分别为40.3、50.0、39.9、61.0、39.2、24.8ng/mL。结论本法操作简单,准确度、灵敏度高,可用于塞来昔布原料药中有关物质的分离检测。     

Effects of acetylsalicylic acid and celecoxib on the N-nitrosodiethylamine induced carcinogenesis in rat liver and esophagus

The effects of nonsteroid antiinflammatory drugs (acetylsalicylic acid and celecoxib) on N-nitrosodiethylamine-induced carcinogenesis in the liver and esophagus were studied in rats. The inhibitory effect of celecoxib on carcinogenesis was more pronounced (in comparison with acetylsalicylic acid), which manifested in a significantly decreased incidence of neoplastic changes in the liver tissue (from 91.7 to 65.2%), number of tumors in the esophagus (from 4.13 to 2.61 tumor/rat), and in delayed malignization in the liver and esophagus. The incidence of erosions and ulcers of the gastric mucosa was significantly lower after celecoxib treatment. These data indicate that celecoxib inhibits N-nitrosodiethylamine-induced carcinogenesis in the liver and esophagus. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 143, No. 1, pp. 93–96, January, 2007     

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.     

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。     

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.     

Effect of Concomitant Polyethylene Glycol and Celecoxib on Colonic Aberrant Crypt Foci and Tumors in F344 Rats

We investigated whether celecoxib augments the protective effect of polyethylene glycol (PEG) on colonic aberrant crypt foci (ACF) and tumor formation in F344 rats treated with azoxymethane (AOM). Three groups of rats received AOM: I (AOM alone), II (PEG), and III (PEG/celecoxib). PEG reduced the mean number of total ACF per colon from 190 to 141 (P < 0.05; 26% reduction) and ≥4-crypt ACF from 95 to 58 (P < 0.01; 39%). Group III rats had a greater proportion of their ACF distally; whereas transverse colon ACF were reduced ∼50%, distal ACF were reduced by only ∼8% (P < 0.05). Of 13 large bowel tumors, 8 were in Group I, 4 in Group II, and 1 in Group III rats (P = 0.02). Thus in AOM-treated rats celecoxib appeared to enhance the PEG-induced reduction in colonic tumor formation, and in transverse but not distal or whole-colon ACF.     

塞来昔布联合 NK 细胞对裸鼠人肝癌皮下移植瘤生长的影响

目的：观察塞来昔布联合NK细胞对裸鼠人肝癌细胞SMMC-7721皮下移植瘤生长的影响,并探讨其可能的作用机制。方法选择裸鼠40只,制备人肝癌细胞SMMC-7721移植瘤模型,随机分为对照组、NK细胞组、塞来昔布组及联合干预组各10只。 NK细胞组瘤内注射NK细胞悬液0．6 mL,每7 d注射1次,共注射5次;塞来昔布组从接种后第3天起给予塞来昔布100 mg/kg灌胃,每天1次,连续35 d;联合干预组同时给予塞来昔布和NK细胞,给药剂量及途径与单用组相同;对照组灌胃和瘤内注射等量生理盐水。35 d后切取移植瘤组织计算体积,称取瘤质量,并计算抑瘤率;TUNEL法评价肿瘤细胞凋亡情况,免疫组化法检测肿瘤内VEGF、Bax、Bcl-2、caspase-3、Ki-67、NF-κB的阳性表达。结果联合干预组移植瘤体积、肿瘤质量均明显低于单用组（ P均＜0．05）,抑瘤率、凋亡指数明显高于单用组（ P均＜0．05）。联合干预组移植瘤中VEGF、Bcl-2、NF-κB、Ki-67表达明显低于单用组（ P均＜0．05）,Bax、caspase-3表达明显高于单用组（P均＜0．05）。结论塞来昔布联合NK细胞可明显抑制裸鼠人肝癌细胞SMMC-7721皮下移植瘤的生长;其作用机制可能是通过促进凋亡级联通路上Bax、caspase-3的表达,抑制VEGF、Bcl-2、NF-κB、Ki-67的表达而实现。     

塞来昔布对阿戈美拉汀抑制作用的体内外研究

目的：研究塞来昔布对阿戈美拉汀的体内外抑制作用及机制。方法：鼠肝微粒体体外实验得到塞来昔布对阿戈美拉汀的抑制机制,并在大鼠体内得到验证。因此,可通过人肝脏微粒体和重组CYP2C9.1蛋白体外实验预测塞来昔布在人体中对阿戈美拉汀的抑制作用和可能机制。结果：塞来昔布表现出明显抑制作用,该作用在CYP2C9.1中呈现浓度依赖性,而在鼠肝微粒体中表现为非竞争性抑制。结论：塞来昔布联合阿戈美拉汀进行抑郁治疗时可能会对后者产生非竞争抑制,这一作用能允许降低阿戈美拉汀的使用剂量,从而降低不良反应的发生,提高治疗的安全性。