Matousek Josef, Poucková Pavla, Hlousková Daniela, Zadinová Marie, Soucek Josef, Skvor Jirí
Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Libechov 277 21, Czech Republic.
J Control Release. 2004 Feb 10;94(2-3):401-10. doi: 10.1016/j.jconrel.2003.10.025.
Subcutaneous application of bovine RNase A conjugated to HYase (bovine hyaluronidase), polyethylene glycol (PEG) and HYase+PEG resulted in a marked reduction of the width of the spermatogenic layers of the mouse testes. The number of sperms in caput epididymidis was significantly decreased in mice injected with conjugated RNase A. There was not any significant embryotoxic effect of free RNase A even conjugated with HYse, PEG and HYse+PEG. The immunogenicity, expressed in production of antibodies against free RNase A or conjugates with PEG, was very low. However, the immunogenic action of this enzyme conjugated only to HYase was much higher and produced the same immunogenicity as HYase itself. The immunogenic effect of RNase A+HYase conjugate decreased when PEG was joined to this conjugate. The inhibitory effect of RNase A conjugated to HYase, PEG and HYase+PEG on human ML-2 cells studied in vitro, was practically ineffective. On the other side, when RNase A conjugated to HYase or PEG was administered intraperitoneally into the mice bearing human melanoma, the antitumor effect was pronounced.
将与透明质酸酶（牛透明质酸酶）、聚乙二醇（PEG）偶联的牛核糖核酸酶A以及透明质酸酶+PEG进行皮下给药，可显著减小小鼠睾丸生精层的宽度。注射偶联核糖核酸酶A的小鼠附睾头部精子数量显著减少。即使与透明质酸酶、PEG以及透明质酸酶+PEG偶联，游离核糖核酸酶A也未产生任何显著的胚胎毒性作用。以针对游离核糖核酸酶A或与PEG的偶联物产生抗体来衡量的免疫原性非常低。然而，仅与透明质酸酶偶联的这种酶的免疫原性作用要高得多，并且产生与透明质酸酶本身相同的免疫原性。当PEG与核糖核酸酶A+透明质酸酶偶联物结合时，该偶联物的免疫原性作用降低。在体外研究中，与透明质酸酶、PEG以及透明质酸酶+PEG偶联的核糖核酸酶A对人ML-2细胞的抑制作用实际上无效。另一方面，当将与透明质酸酶或PEG偶联的核糖核酸酶A腹腔注射到荷人黑色素瘤的小鼠体内时，抗肿瘤作用显著。
