高效液相色谱法测定异烟肼片中异烟肼及其杂质的含量

目的建立测定异烟肼片剂中异烟肼及其杂质含量的高效液相色谱（HPLC）法。方法色谱柱为Shim-PackVP-OD柱（150mm×4．6mm,5μm）,流动相为甲醇-0．2％多库酯钠水溶液（40：60）,流速为0．8mL／min,紫外检测波长为216nm。结果异烟肼质量浓度在10～1000mg／L范围内与峰面积线性关系良好（r=0．9998,n=6）,平均回收率为99．19％,RSD=1．99％（n=6）;异烟酸质量浓度在4～24mg／L范围内与峰面积呈良好线性关系（r=0．9994,n=6）。结论该法快速、简便、准确、重现性好。     

The influence of dose and <Emphasis Type="Italic">N</Emphasis>-acetyltransferase-2 (NAT2) genotype and phenotype on the pharmacokinetics and pharmacodynamics of isoniazid

Objective This study evaluated the pharmacokinetics of isoniazid (INH) associated with optimal early bactericidal activity (EBA), defined as 90% of the maximum EBA (EBA₉₀) and the influence of N-acetyltransferase-2 (NAT2) subtype on the ability of pulmonary tuberculosis (PTB) patients to reach the identified pharmacokinetic values after INH doses ranging from 0.2 to 10–12 mg/kg body weight. Methods INH serum concentrations and NAT2 subtype were determined during four studies of PTB patients in three of whom the EBA of INH was determined. The relationship of EBA to area under the curve (AUC) and 2-h serum concentrations was examined by exponential regression and fitted curves estimated the and 2-h serum concentrations at which EBA₉₀ was reached. Results EBA₉₀ was reached at an of 10.52 μg/ml per hour and 2-h serum concentrations of 2.19 μg/ml. An of 10.52 μg/ml per hour was reached by all 66 patients receiving a 10–12 mg/kg INH dose and all 21 receiving 6 mg/kg, except 1 of 10 (10%) homozygous fast (FF) acetylators; however, at 5 mg/kg, 4 of 12 (33%) FF and 26 of 27 (96%) heterozygous fast (FS), but all 21 homozygous slow (SS) acetylators did so; and 1 of 3 (33%) FF, 2 of 6 (33%) FS, but all 4 SS acetylators at dose 3 mg/kg. An INH 2-h serum concentration of 2.19 μg/ml was reached by all 66 patients receiving 10–12 mg/kg and all 21 receiving 6 mg/kg, except for 2 (20%) FF acetylators at a dose of 5 mg/kg; however, only 3 (25%) of 12 FF acetylators, but 26 (96%) of 27 FS acetylators, and all 21 SS acetylators reached this concentration; and at a dose of 3 mg/kg, 1 (33%) of 3 FF acetylators, 2 (33%) of 6 FF, but all 4 SS acetylators. Conclusions At a 6 mg/kg dose, all except a minority of FF NAT2 acetylators, achieve an INH and 2-h INH serum concentrations associated with EBA₉₀, as did all 4 SS acetylators receiving 3 mg/kg. Any dose reduction below 6 mg/kg body weight will tend to disadvantage a significant proportion of faster acetylators, but, conversely, SS acetylators require only a 3 mg/kg dose to achieve a satisfactory exposure to INH.     

Exploration of Cold Extrusion for the Preparation of Enteric Minitablets of Isoniazid

The objective of the present work was to formulate the enteric minitablets of isoniazid by cold extrusion method. The minitablets were prepared using isoniazid, hydroxylpropylmethylcellulose phthalate and dibasic calcium phosphate. The minitablets were coated using hydroxypropylmethylcellulose phthalate. Full factorial design was adopted to optimize the formulation. The minitablets showed good flow and acceptable friability. The drug release was resisted in 0.1 N HCl for 2 h from the optimized batch. The optimized batch showed more than 90% of drug release in phosphate buffer in 15 min. Capsules containing rifampicin powder and enteric isoniazid minitablets showed complete drug release in acidic and alkaline media respectively. The process of cold extrusion appears to be an attractive alternative to by-pass the existing patents.     

Low-density poly( -lactide-co-glycolide) foams for prolonged release of isoniazid

Incorporation of the antitubercular drug isoniazid, INH, into low density poly( -lactide-co-glycolide), PLGA, foams of high interstitial void volume prior to high pressure extrusion is shown to prolong the in vitro release of INH. In vitro studies indicate that the duration of INH release can be significantly increased, the early burst dramatically reduced, and variation in replicate samples reduced. Control of the specific gravity and interstitial void volume of the foam is achieved by lyophilization of frozen polymer solutions of specified concentration. The morphology of foams prepared by lyophilization of glacial acetic acid solutions of the polymers produces leaflet or platelet structures. Matrices were prepared by (1) extruding INH impregnated foams previously compacted and ground to 125–180 microns, (2) directly extruding impregnated foams without prior compaction and grinding, and (3) extruding mechanically mixed micronized INH and ground PLGA which had not been prepared as foam. INH release kinetics, analyzed in terms of the Roseman-Higuchi model, confirms that release is diffusion controlled. Diffusion constants for the three methods are 1.2( ±0.1) × 10⁻⁴, 2.1(±0.3) × 10⁻⁴, and 3.2(± 1.6) × 10⁻⁴ cm²/day.     

高效液相色谱法同时测定血浆中异烟肼和乙酰异烟肼

目的:建立同时测定血浆中异烟肼和乙酰异烟肼的高效液相色谱(HPLC)法。方法:血浆样品经10%高氯酸沉淀蛋白后,取上清液直接进样分析。选用LichrospherC18柱(250mm×4.6mm,7μm),流动相为乙腈∶20mmol/L磷酸缓冲液(含2mmol/L庚烷磺酸钠)(2∶98,v/v),检测波长266nm。结果:异烟肼和乙酰异烟肼的血药浓度线性范围分别为0.12～15.89mg/L(r=0.998)和0.13～17.08mg/L(r=0.997),提取回收率分别为78.1%～95.6%和83.4%～94.4%,日内和日间变异系数(RSD)均<10%。结论:本研究建立的方法简便、准确,能满足药代动力学研究中生物样品测定的要求。     

中国汉族结核病患者 N-乙酰基转移酶2基因型与药物性肝损伤以及抗结核疗效的关系

目的：了解中国汉族结核病患者 N-乙酰基转移酶（NAT2）基因型与异烟肼引起的药物性肝损伤及与抗结核疗效的关系。方法初治结核病患者108例。采用 PCR 直接测序法对 NAT2基因7个单核苷酸多态性位点分析比对,判定 NAT2基因型。分析基因型与药物性肝损伤的关系,比较基因型与抗结核疗效的关系。统计学处理采用χ2检验。结果108例结核病患者中,中间代谢型59例（54．63％）,快代谢型36例（33．33％）,慢代谢型11例（10．19％）,超快代谢型2例（1．85％）。20例药物性肝损伤患者中快代谢型2例,慢代谢型5例,中间代谢型13例。快代谢型患者发生药物性肝损伤可能性较低（OR ＝0．176,95％CI ：0．038～0．809,P ＝0．014）,慢代谢型患者易发生药物性肝损伤（OR ＝4．556,95％CI ：1．231～16．854,P ＝0．044）。NAT2*4／*6A 基因型（OR ＝7．741,95％CI ：2．653～22．586,P ＜0．01）和 NAT2*6A／*6A 基因型（OR＝15．353,95％CI ：1．506～156．552,P ＝0．020）患者易发生药物性肝损伤,NAT2*4／*4型患者则不易发生药物性肝损伤（OR ＝0．176,95％CI ：0．038～0．809,P ＝0．014）。58例痰菌阳性的患者,治疗2个月后仍痰菌阳性12例,其中快代谢型患者8例,疗效差（OR＝7．200,95％CI ：1．794～28．900,P ＝0．008）。结论中国汉族结核病患者中 NAT2基因型以中间代谢型为主。NAT2慢代谢型发生药物性肝损伤的风险较高。NAT2*6A 为药物性肝损伤高风险等位基因,而 NAT2*4／*4则可能为保护性基因型。快代谢型患者早期的疗效较差。     

耐异烟肼结核分枝杆菌临床分离株耐药相关基因突变研究

目的阐明结核分枝杆菌耐异烟肼临床分离株katG、inhA、ahpC、kasA及oxyR基因突变特点。方法对144株结核分枝杆菌临床分离株(耐异烟肼菌株101株;异烟肼敏感株43株)的katG、inhA、kasA、ahpC及oxyR基因进行DNA片断扩增及DNA序列分析,与GeneBank中结核分枝杆菌标准序列进行比较。结果(1)耐异烟肼菌株中未发现katG完全缺失,81株耐药株(80.2%)katG存在点突变、缺失或插入,其中16个突变位点未见报道;39株(38.6%)耐药株第315位点突变,低耐药菌株(1μg/ml)第315位点突变率显著高于高耐药菌株(10μg/ml;χ2=9.31,P<0.05);58株(57.4%)耐药株第463位点突变。23株(53.3%)敏感株第463位点突变。(2)5株(4.9%)耐药株inhA发生突变。敏感株inhA无突变。(3)3株(2.9%)耐药株ahpC发生突变。敏感株ahpC无突变。(4)17株(16.8%)耐药株kasA发生突变。敏感株中3株菌株Gly312Ser突变。(5)在全部菌株中未发现oxyR基因突变。(6)综合本项研究中各基因的突变情况,共有91株耐异烟肼菌株发生与异烟肼耐药相关的突变。结论本项研究进一步证实了结核分枝杆菌耐异烟肼与katG、inhA、ahpC及kasA基因突变之间的关系,并且提示还有其他机制参与异烟肼耐药。     

Metabolic studies on the possible mode of action of isoniazid tumorigenicity

Summary Data on tumorigenicity and mutagenicity of INH show that INH is tumorigenic in mice but not in rats. The metabolic studies on the two species denote that rats are rapid inactivators whereas mice are slow inactivators of INH. Rats are also resistant to the immediate inhibitory effect of INH on DNA biosynthesis. Using Ames test it was observed that INH is mutagenic to salmonella typhimurium strains TA 100 and 1535 and this effect is abolished in presence of 59 mixture. In vivo and in vitro studies on INH interaction with macromolecules reveal that there is a greater interaction with RNA than with DNA and the site of interaction is the cytidine and deoxycytidine, respectively. A preliminary study is undertaken to see if healed TB cases have a higher risk for cancer. It is found that cancer incidence in this group is higher as compared to noncancer patients.     

结核分枝杆菌临床分离株异烟肼耐药性四种测定方法的比较

目的探讨噬菌体生物扩增(PhaB)、Bactec-960、最低抑菌浓度(MIC)和基因芯片方法在结核分枝杆菌(MTB)异烟肼(INH)耐药性测定中的应用价值。方法应用4种方法检测167株MTB临床分离株INH耐药性，并比较测定结果的敏感性、特异性和准确性。结果Bactec-960检测敏感株111株，耐药株56株。若以Bactec-960测定结果为判断标准，则PhaB的敏感性、特异性、阳性预测值、阴性预测值和准确性分别为96．4％、96．4％、93．1％、98．2％和96．4％；MIC的敏感性、特异性、阳性预测值、阴性预测值和准确性分别为92．9％、99．1％、98．1％、96．5％和97．0％；基因芯片的敏感性、特异性、阳性预测值、阴性预测值和准确性分别为83．9％、96．4％、92．2％、92．2％和92．2％。若以MIC测定耐药性结果为判断标准，则PhaB检测INH耐药性的敏感性、特异性、阳性预测值、阴性预测值和准确性分别为100％、95．6％、91．4％、100％和97．0％；Bactec-960的敏感性、特异性、阳性预测值、阴性预测值和准确性分别为98．1％、96．5％、92．9％、99．1％和97．0％；基因芯片的敏感性、特异性、阳性预测值、阴性预测值和准确性分别为88．7％、96．5％、92．2％、94．8％和94．0％。结论PhaB检测INH耐药性具有很高的敏感性和特异性，与Bactec-960及MIC的符合率很高，只需3d时间，且操作简便、不需特殊仪器设备，可作为MTB的INH耐药性快速筛选方法。MIC测定INH耐药性结果与Bactec-960符合率较高，是否用于INH耐药性测定的判断标准有待进一步研究。基因芯片检测INH耐药性敏感性偏低，只能作为参考方法。     

补肺活血胶囊联合四联抗结核疗法治疗结核性胸膜炎的临床研究

目的观察补肺活血胶囊联合四联抗结核疗法治疗结核性胸膜炎的疗效。方法选取2015年4月—2020年9月在新乡医学院第一附属医院诊治的96例结核性胸膜炎患者为研究对象,随机分为对照组(48例)和治疗组(48例)。对照组患者口服利福平片,0.6 g/次,1次/d;异烟肼片,0.3 g/次,1次/d;吡嗪酰胺片,1.25 g/次,1次/d;盐酸乙胺丁醇片,0.75g/次,1次/d。治疗组在对照组的基础上口服补肺活血胶囊,4粒/次,3次/d。两组患者连续治疗6周。观察两组患者临床疗效,比较治疗前后两组患者胸水吸收时间、住院时间、临床症状缓解时间、胸膜厚度,炎症因子白介素-6(IL-6)、C反应蛋白(CRP)和肿瘤坏死因子-α(TNF-α)水平及肺功能第1秒用力呼气容积(FEV1)和用力肺活量(FVC)。结果治疗后,治疗组的总有效率明显高于对照组(91.67%vs 72.92%,P0.05)。治疗后,治疗组的胸水吸收时间、住院时间、临床症状缓解时间明显短于对照组,胸膜厚度小于对照组(P0.05)。治疗后,两组患者IL-6、CRP、TNF-α水平均较治疗前显著下降(P0.05),且治疗组明显低于对照组(P0.05)。治疗后,两组患者FEV1、FVC均较治疗前升高(P0.05),且治疗组高于对照组(P0.05)。结论补肺活血胶囊联合四联抗结核疗法治疗结核性胸膜炎,可有效阻止疾病进展,缓解临床症状,促进肺功能改善,疗效可靠。