CA77.1

CA77.1 is an orally active small-molecule derivative of 7-chloro-3-(p-tolyl)-2H-benzo[b][1,4]oxazine (AR7) and can act as an activator of chaperone-mediated autophagy (CMA)^[1]. CA77.1 exerts neuroprotective effects and is being developed for the treatment of various chronic CNS diseases including Alzheimer's disease, Parkinson’s disease and retinal degeneration^[2].

In vitro, CA77.1 (12μM) pretreated murine microglial BV2 cells for 12h followed by LPS treatment. CA77.1 decreased LPS-induced iNOS and COX-2 protein expression, lowered the proinflammatory factors NO and IL-6 and reduced the mRNA expression of iNOS, COX-2, IL-6, and IL-1β without affecting cell viability^[3]. CA77.1 (20µM) treatment on mouse neuroblast Neuro-2a (N2a) cells for 24h promoted the degradation of GAPDH (a canonical substrate of CMA) and augmented Na⁺ /K⁺ -ATPase β1 subunit elimination, accompanied by decreased Na⁺ /K⁺ -ATPase activity and increased concentration of intracellular Na⁺, without mRNA levels changed^[4]. CA77.1 (0-30μM) treatment on PK-15 porcine kidney cells expressing foot-and-mouth disease virus (FMDV) non-structural protein 3D for 24h effectively diminished FMDV 3D polymerase levels and suppressesd FMDV replication via activation of the CMA pathway^[5].

In vivo, CA77.1 (10mg/kg/day) was intraperitoneally injected into intracerebral hemorrhage (ICH) mice models for six consecutive days. CA77.1 administration reversed ICH induced augmentation of A1 reactive astrocytes, myelin damage, neuronal death, and neurobehavioral disorders^[6]. CA77.1 (10mg/kg) was administered intraperitoneally into mitochondrial toxin BDE-47-exposed mice with cognitive impairment every three days for 4 weeks. CA77.1 mitigated BDE-47-induced cognitive impairment in mice by suppressing ferroptosis, preventing synaptic loss in mice hippocampi and rescuing hippocampal neuronal structural atrophy^[7].

References:
[1] Bourdenx M, Martín-Segura A, Scrivo A, et al. Chaperone-mediated autophagy prevents collapse of the neuronal metastable proteome.Cell. 2021 May 13;184(10):2696-2714.e25.
[2] Wu J, Xu W, Su Y, et al. Targeting chaperone-mediated autophagy in neurodegenerative diseases: mechanisms and therapeutic potential.Acta Pharmacol Sin. 2025 Apr;46(4):816-828.
[3] Wu J, Han Y Y, Xu H, et al. Deficient chaperone-mediated autophagy facilitates LPS-induced microglial activation via regulation of the p300/NF-κB/NLRP3 pathway. Sci Adv. 2023 Oct 6;9(40):eadi8343.
[4] Zhang Y X, Wu H H, Zhang Q, et al. LAMP2A-mediated neuronal hyperexcitability by enhancing NKAβ1 degradation underlies depression-induced allodynia. Cell Rep. 2025 Apr 22;44(4):115489.
[5] Ren M, Zhou H Q, Wu J E, et al. Heat shock protein A1 inhibits the replication of foot-and-mouth disease virus by degrading viral RNA polymerase 3D through chaperone-mediated autophagy. J Virol. 2025 May 20;99(5):e0016825.
[6] Zheng Y, Peng L, Jiang G N, et al. Activation of chaperone-mediated autophagy exerting neuroprotection effect on intracerebral hemorrhage-induced neuronal injury by targeting Lamp2a. Exp Neurol. 2024 Dec:382:114986.
[7] Yuan Q, Wang M W, Zhang Z X, et al. The ameliorative effects of melatonin against BDE-47-induced hippocampal neuronal ferroptosis and cognitive dysfunction through Nrf2-Chaperone-mediated autophagy of ACSL4 degradation. Ecotoxicol Environ Saf. 2025 Jan 15:290:117542.

CA77.1是一种口服活性小分子，为7-氯-3-(对甲苯基)-2H-苯并[b][1,4]氧嗪（AR7）的衍生物，可作为伴侣介导自噬（CMA）的激动剂^[1]。CA77.1具有神经保护作用，被开发用于治疗各种慢性中枢神经系统疾病，包括阿尔茨海默病、帕金森病和视网膜变性^[2]。

在体外实验中，CA77.1（12μM）预处理小鼠小胶质BV2细胞12h后再以LPS刺激，CA77.1抑制了LPS诱导的iNOS和COX-2蛋白表达，降低促炎因子NO和IL-6水平，并下调iNOS、COX-2、IL-6和IL-1β的mRNA表达，且不影响细胞活力^[3]。CA77.1（20μM）作用于小鼠神经母细胞Neuro-2a（N2a）细胞24h，CA77.1促进CMA经典底物GAPDH的降解，并增强Na⁺/K⁺-ATPase β1亚基的清除，伴随Na⁺/K⁺-ATPase活性下降及胞内Na⁺浓度升高，而相应mRNA水平无变化^[4]。CA77.1（0-30μM）处理表达口蹄疫病毒（FMDV）非结构蛋白3D的PK-15猪肾细胞24小时后有效降低了细胞中FMDV 3D聚合酶水平，并通过激活CMA途径抑制了FMDV复制^[5]。

在体内实验中，CA77.1（10mg/kg/day）连续6天腹腔注射脑出血（ICH）小鼠模型，可逆转ICH诱导的A1反应性星形胶质细胞增多、髓鞘损伤、神经元死亡及神经行为障碍^[6]。CA77.1（10mg/kg）每3天腹腔给药一次，持续4周，可缓解线粒体毒素BDE-47暴露所致的小鼠认知障碍，通过抑制铁死亡、阻止海马突触丢失及挽救海马神经元结构萎缩发挥作用^[7]。