乙二胺四甲叉膦酸骨靶向性异烟肼载体的设计和在脊柱结核动物模型中释放的研究

It is well known that spinal tuberculosis focus can be resected directly with surgery，But tuberculosis bacteria is unresectable. Residues focus of postoperative patients with Mycobacterium tuberculosis, especially placed with internal fixation could be made of the adherence of Mycobacterium tuberculosis on the metal bed. Along with bones special composition and physiological activity, and half-life of anti-TB drug is shorter and the organization permeability of itself is not strong it is difficult for therapeutic anti-tuberculosis drugs to efficient transport to the site of necessary place by the general route, which has become an important cause of postoperative patients for the recurrence of the spinal tuberculosis.Matrix of hydroxyapatite is main components of human bone , and it has become a important target of bone to the ideals of the specificity of binding sites.Multi-phosphoric acid compounds on bone matrix has high affinity, Therefore we came up with a hypothesis that Multi-phosphate can be used as a specific bone targeting carrier in combination with anti-TB drugs and we had designed a bone-targeting of ethylene diamine four methylene phosphonic acid as a carrier of isoniazid, made it continuous suppled with anti-TB drugs to bones from the peripheral blood for improving concentration of them by this specific bone targeting carrier.At the same time we had produced a isoniazid controlled release agents and directly embedded this anti-TB drugs controlled release agents in the region of surgery at the end of operation to remains the anti-TB drugs concentration of bones in the long term remaining above effective bactericidal concentrations,for effectively improving the sterilization drug performance.By isotope labeling the isoniazid for tracking the disstribution of anti-tuberculosis drugs in the bertebra of healthy rabbits could effectively avoided differences of concentration of antituberculosis drugs due to bias of sample collection,Then we will continue to use this tracking technology and validate this specific bone targeting carrier in animal models of spinal tuberculosis in vivo, test the concentration of anti-TB drugs in rabbit spinal tuberculosis model after using this specific bone isoniazid targeting vectors and controlled release agents in long and short term, With a view to improving the treatment of spinal tuberculosis and maximizing the germicidal efficacy of anti-tuberculosis drugs, effectively reduce postoperative recurrence of tuberculosis of the spine.

外科手术可以直接切除脊柱结核病灶，但不能完全清除结核菌。而手术放置内固定则有可能成为术区残留结核菌粘附的金属床，加上骨骼特殊的成分和生理活性，以及抗结核药物自身半衰期较短和组织渗透性差的特点，治疗药物按一般给药途径难以有效转运到作用部位，成为了脊柱结核术后复发的一个重要原因。羟基磷灰石是人类骨基质的主要成分同时也成为了骨靶向理想的特异性结合位点，多元磷酸类化合物对骨基质具有高度的亲和性。因此本题提出了多元磷酸盐可以作为与抗结核药物结合的特异性骨靶向性载体的假说，设计了一种具有骨靶向性的乙二胺四甲叉膦酸作为异烟肼载体，从外周血液特异性地将抗结核药物输送到骨骼的结核病灶中，并直接在术区埋置抗结核药物缓释剂，使骨骼中的抗结核药物浓度长期保持在有效杀菌药物浓度，最后在脊柱结核动物模型病灶内进行浓度监测的验证。从而有效地提高脊柱结核的疗效，减少脊柱结核术后的复发，为骨结核的治疗提供理论依据。

外科手术可以直接切除脊柱结核病灶，但不能完全清除结核菌。在放置内固定后术区残留结核菌有可能成为结核菌粘附的金属床，加上骨骼特殊的成分和生理活性，以及抗结核药物自身半衰期较短和组织的渗透性差，治疗药物按一般给药途径难以有效转运到作用部位，成为了脊柱结核术后复发的一个重要原因。羟基磷灰石是人类骨基质的主要成分，同时也是骨靶向较理想的特异性结合位点。.本题提出了多元磷酸盐可以作为与抗结核药物的特异性骨靶向性载体的假说，并设计了一种具有具有骨靶向性的乙二胺四甲叉膦酸作为异烟肼载体，从外周血液特异性地将抗结核药物输送到骨骼的结核病灶中，同时直接在术区埋置抗结核药物缓释剂，使骨骼中的抗结核药物浓度长期保持在有效杀菌药物浓度，并在脊柱结核动物模型病灶内进行浓度监测的验证。从而有效地提高脊柱结核的疗效，减少脊柱结核术后的复发，为骨结核的治疗提供理论依据。.通过研究，获得的主要成果有：①制作出能在脊柱结核病灶术区内持续长时间释放抗结核药物的缓释载体；②本题所采用质谱条件能一次性进行检测全部一线五种抗结核药物浓度；③对实验动物进行造模干预后模拟人体抗结核治疗的过程，实验动物病灶内的药物浓度并与人体抗痨治疗后药物浓度有一定差异，实验动物体内埋置异烟肼缓释微球可保持局部抗结核药物的高浓度水平；④采用同位素示踪技术，用放射性核素锝-99标记异烟肼，同时验证了聚乳酸异烟肼微球、羟基磷灰石异烟肼微球及异烟肼原药44、48、52天透析液的结核菌培养抑制效果；⑤使用聚乳酸-羟基乙酸共聚物作为缓释载体搭载利福平构建抗结核缓释微球，并完成复合材料的体内缓释实验、生物安全性实验和体内成骨实验；⑥设计 ALN-PLGA 纳米粒装载异烟肼的靶向制剂，完成其载体毒性试验和体内外靶向性试验。.本项目完成3篇SCI论文，1篇中文核心期刊论文，3篇2014年SICOT会议录用论文大会发言，培养博士生3人，硕士生2人。