Antitumor agent-81 (compound 5 a) is a low cytotoxic P62-RNF168 agonist that promotes the interaction between P62 and RNF168. Antitumor agent-81 induces RNF168-mediated reduction of H2A ubiquitination and impairs homologous recombination-mediated DNA repair. Antitumor agent-81 also inhibited mouse xenograft tumor growth in a dose-dependent manner.

Physicochemical Properties

Molecular Formula	C19H19N7O3
Molecular Weight	393.399262666702
Exact Mass	393.154
CAS #	2765180-17-2
PubChem CID	163214076
Appearance	Off-white to light yellow solid powder
LogP	1.8
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	5
Heavy Atom Count	29
Complexity	530
Defined Atom Stereocenter Count	0
InChi Key	SWULRQWLALOGFO-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H19N7O3/c1-27-15-7-11(8-16(28-2)17(15)29-3)26-9-14(24-25-26)10-4-5-13-12(6-10)18(20)23-19(21)22-13/h4-9H,1-3H3,(H4,20,21,22,23)
Chemical Name	6-[1-(3,4,5-trimethoxyphenyl)triazol-4-yl]quinazoline-2,4-diamine
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
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	The most prevalent cancer types' cells show antiproliferative activity when exposed to antitumor agent-81 (0–20 µM; 72 h)[1]. In HCT-116 cells, antitumor agent-81 (1, 2, 5 µM; 24 h) impairs HR (homologous recombination)-mediated DSB repair[1]. In HCT-116 cells, antitumor agent-81 tethers the relationship between P62 and RNF168[1]. In HCT-116 cells, antitumor agent-81 inhibits RNF168's catalytic activity as well as RNF168 E3 ligase activity[1].
ln Vivo	Tumor volumes are suppressed in mice by antitumor agent-81 (5, 10 mg/kg; ip; once every three days for 22 days) in a dose-dependent manner[1]. 1.19 Antitumor agent-81 pharmacokinetic parameters in female BALB/c nude mice[1]. IP (5 mg/kg) 16.17 26.10 215.69 19647.83 IP (10 mg/kg) 31.00 39.87 408.18 16554.30 IP (20 mg/kg) 22.10 52.23 1003.58 14669.81 Dose T1/2 (h) Cmax (ng/mL) AUC0-t (h㻿ng/mL) CL ((mL/h)/kg)
Cell Assay	Cell Proliferation Assay[1] Cell Types: A549, HCT-116, A375, HeLa, HepG2, MCF-7, MDA-MB-231, MGC-803, U2OS and MCF-10A cells Tested Concentrations: 0 -20 µM Incubation Duration: 72 h Experimental Results: Inhibited A549, HCT-116, A375, HeLa, HepG2, MCF-7, MDA-MB-231, MGC-803, U2OS and MCF-10A cells, with IC50 values of 1.18, 0.36, 1.93, 2.48, 19.61, 2.79, 1.17, 1.88, 3.64 and 6.98, respectively. Cell Cycle Analysis[1] Cell Types: HCT-116 cells Tested Concentrations: 1, 2, 5 µM Incubation Duration: 24 h Experimental Results: Induced a G2/M arrest but no significant accumulation of sub-G1 phase. Compromised both HR(homologous recombination)- and NHEJ (non-homologous end joining)- mediated DSB repair in a dose-dependent manner, but HR was the more severely impacted process .
Animal Protocol	Animal/Disease Models: Female BALB/c nude mice (xenograft tumor model)[1]. Doses: 5, 10 mg/kg Route of Administration: intraperitoneal (ip) injection; single every 3 days for 22 days Experimental Results: Suppressed tumor growth by promoting apoptosis in xenografted tumorigenesis. Animal/Disease Models: Female BALB/c nude mice[1]. Doses: 5, 10, 20 mg/kg Route of Administration: intraperitoneal (ip) injection; single Experimental Results: demonstrated plasma concentration peaked 1 h after administration. demonstrated a relatively high maximum concentration (Cmax= 52.23 ng/mL) and exposure (AUC0-t= 1003.58 h·ng/mL) at a dose of 20 mg/kg.
References	[1]. A 1,2,3-Triazole Derivative of Quinazoline Exhibits Antitumor Activity by Tethering RNF168 to SQSTM1/P62. J Med Chem. 2022 Nov 4.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.5419 mL	12.7097 mL	25.4194 mL
	5 mM	0.5084 mL	2.5419 mL	5.0839 mL
	10 mM	0.2542 mL	1.2710 mL	2.5419 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.