目的：在進(jìn)行聚乙二醇化重組人粒細(xì)胞集落刺激因子（PEG30-rhG-CSF）大鼠和Beagle犬重復(fù)給藥毒性試驗(yàn)時(shí)，觀察給藥后動物血清中抗PEG30-rhG-CSF抗體的產(chǎn)生和抗體的中和活性，為非臨床安全性評價(jià)的確切性以及臨床給藥周期提供依據(jù)。方法：大鼠分為賦形劑對照組、PEG30-rhG-CSF 0.75、3.0、12 mg / kg組，sc給藥，隔日1次，連續(xù)2周；Beagle犬分為賦形劑對照組、PEG30-rhG-CSF 0.25、1.0和4.0 mg / kg，sc，每周1次，連續(xù)4周。采用ELISA法檢測動物血清中抗PEG30-rhG-CSF的抗體，采用NFS-60細(xì)胞／MTT比色法檢測抗體的中和活性。結(jié)果：大鼠在給藥期和恢復(fù)期各劑量組動物血清中均未檢測到抗PEG30-rhG-CSF的結(jié)合抗體。Beagle犬在給藥的第1周和第2周，未檢測到結(jié)合抗體，但在第4周時(shí)，低、中和高3個(gè)劑量組各有5只（5/6）的動物血清中檢測到結(jié)合抗體，而抗體滴度與劑量無明顯相關(guān)性?；謴?fù)4周后，僅高劑量組1只（1/2）動物出現(xiàn)抗體，且抗體滴度呈下降趨勢。另外，在給藥期和恢復(fù)期，血清中所產(chǎn)生的抗體均無中和PEG30-rhG-CSF的活性。結(jié)論：大鼠重復(fù)給予PEG30-rhG-CSF的毒性試驗(yàn)中，所有動物血清中均未檢測到結(jié)合抗體和中和抗體；Beagle犬用藥組的絕大部分動物血清中出現(xiàn)抗PEG30-rhG-CSF的抗體，但所產(chǎn)生抗體無中和活性。

Objective：To investigate the anti-pegylated recombinant human granulocyte colony stimulating factor（PEG30-rhG-CSF） antibody formation and neutralizing antibody activity of PEG30-rhG-CSF in repeated-dose toxicity studies in rats and Beagle dogs. Methods: The rats received sc injection of PEG30-rhG-CSF at doses of 0 (placebo), 0.75, 3.0，and 12 mg / kg (every other day dosing for up to 2 weeks) and Beagle dogs at doses of 0 (placebo), 0.25, 1.0，and 4.0 mg / kg (weekly dosing for up to 4 weeks), respectively. Anti-PEG30-rhG-CSF antibody in serum was measured using ELISA and the neutralizing antibody activity was determined by cell-based bioassay. Results: No anti-PEG30-rhG-CSF responses were observed in rats during the experiment. The antibody titers in serum samples were determined in dogs during the treatment and recovery period, but no neutralizing antibodies were found. There was no apparent dose-response in either incidence of antibody positivity or titer of antibody development in dogs, and titers sharply declined at the end of the recovery period. Conclusion: Antibodies to PEG30-rhG-CSF were not detected in the rats. The dogs have developed binding antibodies, but no neutralizing potency.

“重大新藥創(chuàng)制”科技重大專項(xiàng)（Z20082X09305-005）；天津創(chuàng)新藥物安全評價(jià)技術(shù)平臺建設(shè)。