抗结核药缓释涂层在兔脊柱结核模型体内的释药性能及组织分布研究

Study on tissue distribution of HRZ coating material of sustained-release anti-tuberculosis drug in rabbit spinal tuberculosis model

中文关键词:  脊柱结核  释药性能  组织分布  异烟肼  利福平  吡嗪酰胺

英文关键词:Spinal tuberculosis  Drug release  Tissue distribution  Isoniazid  Rifampicin  Pyrazinamide

中文摘要:

  【摘要】 目的：观察抗结核药缓释涂层在兔脊柱结核模型体内的释药性能及组织分布特性。方法：选取新西兰大白兔（雌雄不限）体重3.00±0.25kg，通过椎体钻孔、浸注结核杆菌法构建并筛选脊柱结核模型120只，按“异烟肼（INH，H）、利福平（RFP，R）和吡嗪酰胺（PZA，Z）”给药剂型及途径不同，将模型分为4组，每组各30只：A组，缓释材料组，病灶局部置入缓释抗结核药缓释涂层（300mg/kg）+自体髂骨；B组，局部给药组，病灶局部置入HRZ药物晶体（H，12.5mg/kg；R，12.5mg/kg；Z，25.0mg/kg）；C组，灌喂给药组，每日固定时间灌喂给药（H，12.5mg/kg/d；R，12.5mg/kg/d；Z，25.0mg/kg/d）；D组，假手术灌药组，每天固定时间给药（剂量：H，12.5mg/kg/d；R，12.5mg/kg/d；Z，25.0mg/kg/d）。四组单次给药剂量相同，其中A、B两组单次给药，C、D两组研究期间连续多次给药。采用高效液相色谱法测定给药后（7d、14d、28d、84d）各组静脉血、腰大肌以及骨组织（病灶椎体）中H、R及Z的药物浓度，绘制药物浓度-时间曲线，观察其释药性能及组织分布特性。结果：4组单次给药剂量相同，A、B两组HRZ各药的给药总剂量均显著小于C、D两组（P<0.01）。A组术后（7d、14d、28d、84d）腰大肌及骨组织中H、R及Z的药物浓度均显著高于C、D两组（P<0.01）。A组术后（7d、14d、28d、84d）静脉血中H、R及Z药物浓度均极低或检测不到（P>0.05）。A组在腰大肌及骨组织中的药物浓度-时间曲线平缓，无突释现象出现。A组腰大肌H（14.22±1.07μg/ml）、R（12.66±1.12μg/ml）、Z（28.93±1.30μg/ml）的浓度和骨组织中H（12.46±0.29μg/ml）、R（10.34±0.32μg/ml）及Z（26.21±0.82μg/ml）的浓度在给药后第7d时最高，至给药后第84d时H、R及Z的浓度降至最低，此时病灶骨组织中H（6.69±1.42μg/ml）、R（6.28±0.77μg/ml）及Z（19.88±0.90μg/ml）的浓度在各检测时间点均≥10倍的最低抑菌浓度。B组术后7d时腰大肌H（8.19±1.98μg/ml）、R（16.87±3.03μg/ml）、Z（91.18±11.12μg/ml）及骨组织H（5.70±1.25μg/ml）、R（13.06±1.26μg/ml）、Z（79.00±8.68μg/ml）中的药物浓度较高，此后急剧下降，至术后28d时腰大肌及骨组织中已检测不到药物（H、R及Z）浓度。B组静脉血中的药物（H、R及Z）浓度极低或检测不到。C、D两组静脉血、腰大肌及骨组织在各检测时间点均可测得药物（H、R及Z）浓度。C组及D组骨组织中药物（H、R及Z）的浓度均维持在相对平稳的较低水平。结论：缓释HRZ材料在兔脊柱结核病灶局部释药性能满意，组织分布特征为病灶局部高药物浓度而外周低药物浓度。

英文摘要:

  【Abstract】 Objectives: To observe the drug release and tissue distribution characteristics of anti-tuberculosis drug′s sustained-release coating in rabbit spinal tuberculosis model. Methods: New Zealand white rabbits of either sex weighing 3.00±0.25kg were selected. A total of 120 spinal tuberculosis models were constructed and screened by vertebral body drilling and infusion of Mycobacterium tuberculosis. According to different drug delivery dosage forms and ways of the "isoniazid (INH, H), rifampicin (RFP, R), and pyrazinamide (PZA, Z)", the rabbit model of spinal tuberculosis were randomly divided into four groups of 30 rabitts: group A, sustained-releasematerial group, with local implantation of sustained-release of HRZ material (300mg/kg)+ autogenous iliac bone; group B, local administration group, with local implantation of HRZ drug crystals (H,12.5mg/kg; R, 12.5mg/kg; Z, 25.0mg/kg); group C, intragastrically fed group, fed intragastricallyat fixed time daily (H, 12.5mg/kg/d; R, 12.5mg/kg/d; Z, 25.0mg/kg/d); group D, sham operation perfusion group, with daily fixed time perfusion (dose: H, 12.5mg/kg/d; R, 12.5mg/kg/d; Z, 25.0mg/kg/d). All groups were given the same dose in a single dose. Group A and B were given single dose, while group C and D were given multiple doses consecutively. Five rabbits were randomly selected from each group at each time point, and drug concentrations of H, R and Z in venous blood, psoas major muscle and bone tissue of each group were determined by high performance liquid chromatography after administration(7d, 14d, 28d and 84d). Drug concentration-time curves were drawn to observe drug release properties and tissue distribution characteristics. Results: The single dose of the four groups was the same, and the total dose of HRZ in groups A and B was significantly lower than that in groups C and D(P<0.01). The drug concentrations of H, R and Z in psoas major muscle and bone tissue in group A were significantly higher than those in groups C and D(P<0.01). In group A, the concentrations of H, R and Z in venous blood were very low orundetected(P>0.05). In group A, the drug-time curve in psoas major muscle and bone tissue was smooth without sudden release. In group A, the concentration of H(14.22±1.07μg/ml), R(12.66±1.12μg/ml), and Z(28.93±1.30μg/ml) in psoas major muscle and bone tissue H(12.46±0.29μg/ml), R(10.34±0.32μg/ml), and Z(26.21±0.82μg/ml) reached the highest on the 7th day after administration, and the lowest on the 84th day after administration. At this time, the concentrations of H(6.69±1.42μg/ml), R(6.28±0.77μg/ml), and Z(19.88±0.90μg/ml) in the bone tissue of the lesion were more than 10 times of the minimum inhibitory concentration(MIC) at each detection time point. The drug concentrations in the psoas major muscle of H(8.19±1.98μg/ml), R(16.87±3.03μg/ml), and Z(91.18±11.12μg/ml) and bone tissue of H(5.70±1.25μg/ml), R(13.06±1.26μg/ml), and Z(79.00±8.68μg/ml) in group B were higher at 7d after surgery, and decreased sharply, which were undetectable in both psoas major muscle and bone tissue 28d after surgery. In group B, the concentration of drugs(H, R, and Z) in venous blood was very low or could not be detected. The concentration of drugs(H, R, and Z) in venous blood, psoas major muscle and bone tissue of C and D groups could be measured at each detection time point.The concentrations of drugs (H, R, and Z) in bone tissue of groups C and D were maintained at relatively stable and low levels. Conclusions: The sustained-release HRZ material has satisfactory local drug release performance in rabbit spinal tuberculosis lesions, and the tissue distribution characteristics are high local drug concentration and low peripheral drug concentration.