TQB-3804 (TQB3804; EGFR-IN-7; compound 34) is a 4th-generation, oralli bioactive and selective EGFR kinase inhibitor (EGFR-TKI, developed in China and disclosed in patent WO2019015655A1) with potential anticancer activity. It inhibits EGFR (WT) and EGFR (mutant C797S/T790M/L858R) with IC50s of 7.92 nM and 0.218 nM, respectively.

Physicochemical Properties

Molecular Formula	C32H41BRN9O2P
Molecular Weight	694.6048
Exact Mass	693.23
Elemental Analysis	C, 55.33; H, 5.95; Br, 11.50; N, 18.15; O, 4.61; P, 4.46
CAS #	2267329-76-8
Related CAS #	2267329-76-8;
PubChem CID	138911391
Appearance	Light yellow to yellow solid powder
LogP	4.4
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	11
Rotatable Bond Count	8
Heavy Atom Count	45
Complexity	994
Defined Atom Stereocenter Count	0
InChi Key	ZYSKXRAGBGLELB-UHFFFAOYSA-N
InChi Code	InChI=1S/C32H41BrN9O2P/c1-21-18-26(28(44-3)19-27(21)42-12-8-22(9-13-42)41-16-14-40(2)15-17-41)38-32-36-20-23(33)31(39-32)37-25-7-6-24-29(35-11-10-34-24)30(25)45(4,5)43/h6-7,10-11,18-20,22H,8-9,12-17H2,1-5H3,(H2,36,37,38,39)
Chemical Name	5-bromo-4-N-(5-dimethylphosphorylquinoxalin-6-yl)-2-N-[2-methoxy-5-methyl-4-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]phenyl]pyrimidine-2,4-diamine
Synonyms	TQB-3804 TQB3804 TQB 3804
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	For EGFR (WT), EGFR (Δ19del/T790M/C797S), and EGFR (C797S/T790M/L858R), EGFR-IN-7 (10 mM) exhibits a potent inhibitory effect on their respective enzymatic activities, with IC50 values of 7.92 nM, 0.218 nM, and 0.16 nM, respectively[1]. As demonstrated by its IC50 value of 154 nM, EGFR-IN-7 (1 mM) exhibits good selectivity for EGFR (WT) in A431 cells. The Ba/F 3 (EGFRΔ19del/T790M/C797S) triple mutant cells are well-inhibited by EGFR-IN-7 (10 μM-0.508 nM), with an IC50 value of 22 nM[1]. For pEGFR Ba/F 3 (EGFRΔ19del/T790M/C797S) cells, EGFR-IN-7 (10 μM or 100 μM) suppresses phosphorylation activity with an IC50 value of 19 nM[1].
ln Vivo	EGFR-IN-7 (Compound 34; 5-45 mg/kg; oral; daily; for 13 days) exhibited in Ba/F 3 (Δ19del/T790M/C797S)-derived xenografts (CDX) implanted subcutaneously Effective anti-tumor activity) BALB/c nude mouse drug resistance model [1]. EGFR-IN-7 (25 and 50 mg/kg; oral; daily for 3 weeks) effectively reduces tumor growth in the mouse subcutaneous xenograft PC-9 (Δ19del) model [1].
Cell Assay	Cell proliferation experiment [1] Cell Types: A431 cells; Ba/F 3 (EGFRΔ19del/T790M/C797S) Suspension cell Tested Concentrations: 1 mM; 10 μM-0.508 nM Incubation Duration: 3 days Experimental Results: Inhibited cell proliferation.
Animal Protocol	Animal/Disease Models: Ba/F 3 (Δ19del/T790M/C797S)-derived xenograft (CDX) BALB/c nude mice (female, 6-8 weeks, 18-22 g) [1] Doses: 5, 15, 45 mg/kg Route of Administration: Orally, one time/day, for 13 days. Experimental Results: The half-life, plasma and tissue exposure were Dramatically increased, and it had good pharmacokinetic/PK/PK effects in mice. Animal/Disease Models: subcutaneousxenograft PC-9 (Δ19del) model [1] Doses: Days 0-9: 50 mg/kg, Days 10-21: 25 mg/kg Route of Administration: Orally, one time/day, 3 weeks Experimental Results: It has a significant inhibitory effect on tumor growth, has a tumor reducing effect, and shows good anti-tumor efficacy.
References	[1]. Prostaglandin E2 suppresses bacterial killing in alveolar macrophages by inhibiting NADPH oxidase. WO2019015655A1.

Solubility Data

Solubility (In Vitro)

DMSO : ~5 mg/mL (~7.20 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 1.25 mg/mL (1.80 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1.25 mg/mL (1.80 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 1.25 mg/mL (1.80 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.4397 mL	7.1984 mL	14.3968 mL
	5 mM	0.2879 mL	1.4397 mL	2.8794 mL
	10 mM	0.1440 mL	0.7198 mL	1.4397 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.