CA77.1 is a potent, brain-penetrating and orally bioactive activator of chaperone-mediated autophagy (CMA) that displays good pharmacokinetics. CA77.1 is an analogue of AR7 that can increase the expression of lysosomal receptor (LAMP2A) in lysosomes. CA77.1 can improve the behavioral and neuropathological characteristics of the PS19 mouse model and may be utilized in the research/study of Alzheimer's disease (AD).

Physicochemical Properties

Molecular Formula	C16H12CLN3O
Molecular Weight	297.7390
Exact Mass	297.066
CAS #	2412270-22-3
PubChem CID	146439211
Appearance	Off-white to light yellow solid powder
LogP	2.7
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	2
Heavy Atom Count	21
Complexity	371
Defined Atom Stereocenter Count	0
SMILES	ClC1C([H])=C([H])C2C(C=1[H])=NC([H])=C(C1C([H])=C([H])C(=C([H])C=1[H])N([H])C(C([H])([H])[H])=O)N=2
InChi Key	ZQXMPDVGBWOTBY-UHFFFAOYSA-N
InChi Code	InChI=1S/C16H12ClN3O/c1-10(21)19-13-5-2-11(3-6-13)16-9-18-15-8-12(17)4-7-14(15)20-16/h2-9H,1H3,(H,19,21)
Chemical Name	N-[4-(6-chloroquinoxalin-2-yl)phenyl]acetamide
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage.
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro	NIH 3T3 cells exhibit stable expression of the KFERQ-PS-Dendra reporter gene as a result of CA77.1's dose- and time-dependent activation of CMA (0-30 μM; 16 hours). The average of the fluorescence spots within each cell was used to calculate CMA activity [1]. LC3-II expression and autophagic flux in NIH 3T3 cells are unaffected by CA77.1 (20 μM; 6 hours) [1].
ln Vivo	CA-77.1 (10 mg/kg; single dose; oral gavage) exhibits good pharmacokinetics and brain penetration. Cmax, AUClast, Tmax, and T1/2 are, in that order, 3534 ng/g, 8338 h*ng/g, 1 hour, and 1.89 hours [1]. In PS19 mice, previously reported hyperkinesia was normalized to control levels by CA77.1 (oral gavage; 30 mg/kg; 6 months). In the hippocampus, amygdala, and piriform cortex, it also decreased the quantity and percentage of neurons with pathogenic tau conformations. Vehicle-treated PS19 mice showed a decrease in the number of large Iba1-positive cells with rod-dystrophic morphology and an increase in microglia following CA77.1 treatment [1].
Animal Protocol	Animal/Disease Models: 9-month-old CTR or PS19 mice [1] Doses: 30 mg/kg Route of Administration: po (oral gavage); 30 mg/kg; 6-month Experimental Results: Behavior of frontotemporal dementia-related protein toxicity mouse model and neuropathology improved.
References	[1]. Chaperone-mediated autophagy prevents collapse of the neuronal metastable proteome.Cell. 2021 May 13;184(10):2696-2714.e25.

Solubility Data

Solubility (In Vitro)	DMSO : ~5 mg/mL (~16.79 mM) Ethanol : ~1 mg/mL (~3.36 mM)
Solubility (In Vivo)	Solubility in Formulation 1: 2 mg/mL (6.72 mM) in 45% PEG300 +5% Tween-80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.3586 mL	16.7932 mL	33.5864 mL
	5 mM	0.6717 mL	3.3586 mL	6.7173 mL
	10 mM	0.3359 mL	1.6793 mL	3.3586 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.