平阳霉素纳米活性炭的研制及其淋巴靶向性研究

目的 制备淋巴靶向平阳霉素纳米活性炭(PYM-ACH-NP),观察经荷瘤鼠癌周黏膜下注射PYM-ACH-NP后,活性药物平阳霉素在体内各组织器官的药物分布特点,探讨PYM-ACH-NP对淋巴结转移灶的靶向性.方法 制备活性炭纳米微粒(ACH-NP),观察ACH-NP对平阳霉素(PYM)吸附效率.采用改良氯胺T法将放射性核素125I-NaI标记PYM,应用淋巴结高转移癌株U14建立昆明小鼠颈淋巴结转移动物模型,随机分为PYM组和PYM-ACH-NP组,于癌周黏膜下按10mg/kg的剂量分别注射PYM水溶液和PYM-ACH-NP,给药72 h后处死小鼠,取血、心、肝、脾、肺、肾和颈淋巴结,检测各组织器官内的PYM的比放射活性,以考察药物在体内的组织分布情况,对所得数据行t检验.结果 PYM-ACH-NP平均粒径为178 nm,随ACH-NP投入比例的增高,对PYM的吸附率相应增高,10∶1质量比下吸附率达99.38%.癌周黏膜下注射给药后,PYM-ACH-NP组颈淋巴结转移灶内药物比放射活性为(148.72±29.35)cpm/mg远高于PYM组(10.17±2.11)cpm/mg(P＜0.01);PYM-ACH-NP组血、心、肝、脾、肺、肾等器官中的药物比放射活性分别为(2.18±0.39)、(1.19±0.21)、(2.41±0.50)、(1.09±0.24)、(1.95±0.47)、(2.21±0.44)cpm/mg,显著低于PYM组相应各组织中的值(17.22±3.04)、(2.48±0.47)、(6.94±1.38)、(4.12±0.79)、(8.25±2.04)、(18.83±3.89)cpm/mg,差异有统计学意义(P＜0.01).结论 ACH-NP对PYM的吸附率高,PYM-ACH-NP具有良好的淋巴靶向性,经癌周黏膜下给药后,可显著提高淋巴结内的药物浓度,同时减少全身重要脏器的药物分布,降低了药物的全身不良反应.

Abstract：

Objective To explore the possibility of anticancer drug targeting to lymph metastasis using activated charcoal nanoparticles as a drug delivery carrier, the drug distribution in tissues of cervical lymph node metastasis mice model after submucosa adjacent cancer injection of pingyangmycin (PYM ) absorbed in activated charcoal nano-particles ( ACH-NP) and the lymph targeting effect of PYM-ACH-NP.Methods PYM-ACH-NP was prepared by mixed the ACH-NP + PYM + saline and shaken for 20 min. The absorbency of PYM on ACH-NP was evaluated. Cervical lymph node metastasis mice model was established by buccal submucosa implantation of a high lymph metastasis cell line U14 cancer cells (5 × 10 /L). PYM was radiolabeled with 125I by the modified chloramine T method. Thirty Kunming mice models burdened with cervical lymph metastasis were randomly divided into control group, PYM-treated group and PYM-ACH-NP-treated group. The animal in each group was injected with 0. 2 mL saline or corresponding drugs (equal to 10 mg/kg PYM) respectively. The radioactivity of PYM in blood, heart, liver, spleen, lung,kidney and cervical lymph node was detected 72 h after administration. Specific radioactivity of each sample was calculated. Results The average diameter of PYM-ACH-NP was 176 nm. The absorbency of PYM on ACH-NP was increased with the increased ratio of ACH-NP to PYM. In PYM-CH-NP group, specific radioactivity of PYM was significantly higher in cervical lymph node ( 148. 72 ± 29. 35 ) cpm/mg than in PYM group ( 10. 17 ±2. 11) cpm/mg ( P ＜0. 01 ) , meanwhile the specific radioactivity of drug in the blood, heart, liver, spleen, lung and kidney of PYM-CH-NP group was (2. 18 ±0. 39), (1. 19 ±0.21) ,(2.41 ±0.50) , (1.09 ±0.24) , (1.95 ±0.47) and (2.21 ±0.44) cpm/mg respectively, which was significantly lower than in PYM group (17. 22 ± 3. 04) , (2. 48 ± 0. 47 ), (6. 94 ± 1. 38 ), (4. 12 ±0.79), (8. 25 ±2.04), (18. 83 ±3. 89) cpm/mg. The uptake of PYM in the blood, heart, liver,spleen, lung and kidney was greatly decreased in PYM-CH-NP group (P ＜0. 01). Conclusion The activated charcoal nanoparticles has high absorbency of PYM. ACH-NP may serve as a new drug delivery carrier of PYM. Anticancer nanoparticles with the average diameter of 176 nm could deliver anticancer drug to lymph nodes metastasis specifically and decrease drug dosage of other organs. It will be promising in anticancer applications by elevating the therapeutic effect for lymph metastasis and reducing adverse reaction.

Preparation and lymphatic targeting study of pingyangmycin-activated charcoal nanoparticles

Objective To explore the possibility of anticancer drug targeting to lymph metastasis using activated charcoal nanoparticles as a drug delivery carrier, the drug distribution in tissues of cervical lymph node metastasis mice model after submucosa adjacent cancer injection of pingyangmycin (PYM ) absorbed in activated charcoal nano-particles ( ACH-NP) and the lymph targeting effect of PYM-ACH-NP.Methods PYM-ACH-NP was prepared by mixed the ACH-NP + PYM + saline and shaken for 20 min. The absorbency of PYM on ACH-NP was evaluated. Cervical lymph node metastasis mice model was established by buccal submucosa implantation of a high lymph metastasis cell line U14 cancer cells (5 × 10 /L). PYM was radiolabeled with 125I by the modified chloramine T method. Thirty Kunming mice models burdened with cervical lymph metastasis were randomly divided into control group, PYM-treated group and PYM-ACH-NP-treated group. The animal in each group was injected with 0. 2 mL saline or corresponding drugs (equal to 10 mg/kg PYM) respectively. The radioactivity of PYM in blood, heart, liver, spleen, lung,kidney and cervical lymph node was detected 72 h after administration. Specific radioactivity of each sample was calculated. Results The average diameter of PYM-ACH-NP was 176 nm. The absorbency of PYM on ACH-NP was increased with the increased ratio of ACH-NP to PYM. In PYM-CH-NP group, specific radioactivity of PYM was significantly higher in cervical lymph node ( 148. 72 ± 29. 35 ) cpm/mg than in PYM group ( 10. 17 ±2. 11) cpm/mg ( P ＜0. 01 ) , meanwhile the specific radioactivity of drug in the blood, heart, liver, spleen, lung and kidney of PYM-CH-NP group was (2. 18 ±0. 39), (1. 19 ±0.21) ,(2.41 ±0.50) , (1.09 ±0.24) , (1.95 ±0.47) and (2.21 ±0.44) cpm/mg respectively, which was significantly lower than in PYM group (17. 22 ± 3. 04) , (2. 48 ± 0. 47 ), (6. 94 ± 1. 38 ), (4. 12 ±0.79), (8. 25 ±2.04), (18. 83 ±3. 89) cpm/mg. The uptake of PYM in the blood, heart, liver,spleen, lung and kidney was greatly decreased in PYM-CH-NP group (P ＜0. 01). Conclusion The activated charcoal nanoparticles has high absorbency of PYM. ACH-NP may serve as a new drug delivery carrier of PYM. Anticancer nanoparticles with the average diameter of 176 nm could deliver anticancer drug to lymph nodes metastasis specifically and decrease drug dosage of other organs. It will be promising in anticancer applications by elevating the therapeutic effect for lymph metastasis and reducing adverse reaction.