NHWD-870 is a potent, orally bioactive and selective BET family bromodomain inhibitor that binds only to BRD2, BRD3, BRD4 (IC50=2.7 nM) and BRDT. NHWD-870 has a strong tumor suppressor effect and can inhibit the interaction between cancer cells and macrophages.

Physicochemical Properties

Molecular Formula	C29H29N7O
Molecular Weight	491.59
Exact Mass	491.243
CAS #	2115742-03-3
PubChem CID	137448679
Appearance	White to off-white solid powder
LogP	4.1
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	37
Complexity	788
Defined Atom Stereocenter Count	1
SMILES	C12N=CC(C3=C(C)N=NN3C)=CC=1N([C@@H](C1CCOCC1)C1=CC=CC=C1)C1C2=CC2C=NN(C)C=2C=1
InChi Key	YMSCCGUMGIFOSI-GDLZYMKVSA-N
InChi Code	InChI=1S/C29H29N7O/c1-18-28(35(3)33-32-18)22-14-26-27(30-16-22)23-13-21-17-31-34(2)24(21)15-25(23)36(26)29(19-7-5-4-6-8-19)20-9-11-37-12-10-20/h4-8,13-17,20,29H,9-12H2,1-3H3/t29-/m1/s1
Chemical Name	5-(3,5-dimethyltriazol-4-yl)-12-methyl-8-[(S)-oxan-4-yl(phenyl)methyl]-3,8,12,13-tetrazatetracyclo[7.7.0.02,7.011,15]hexadeca-1(9),2(7),3,5,10,13,15-heptaene
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	Melanoma cells (A375) are inhibited by NHWD-870 (0.01-10000 nM) at an IC50 of 2.46 nM[1]. 5 dys; NHWD-870, 0–10,000 nM) inhibited cell growth[1]. NHWD-870 (0–50 nM; 24 hours) suppresses the expression of c-MYC and BRD4 phosphorylation[1]. The hERG channel is mildly inhibited by NHWD-870 (IC50 = 5.4 µM)[1]. Strong effects on apoptosis induction and cell proliferation suppression are demonstrated by NHWD-870[1].
ln Vivo	In mouse models, NHWD-870 (0.75-3 mg/kg; po) exhibits potent anti-tumor properties[1]. Tumor associated macrophages (TAMs) in subcutaneously implanted H526 and A2780 tumors are reduced by NHWD-870. reduced the expression of CSF1 in tumor cells to stop the growth of TAM[1]. distinct cancer situations result in distinct modes of action for NHWD-870. These include: 1) suppressing the proliferation of tumor cells by downregulating the signaling of PDGFRβ, MEK1/2, and STAT1/MYC in tumor cells; 2) suppressing the production of PDGF in tumor cells and the signaling of PDGFRβ and MEK1/2 in endothelial cells in tumor cells to prevent tumor angiogenesis. In xenograft mouse models of triple negative breast cancer, ovarian cancer, and small cell lung cancer, NHWD-870 has strong tumor suppression efficacies[2].
Cell Assay	Cell Viability Assay[1] Cell Types: H526, A2780, ES-2, and MDA-MB231 cells Tested Concentrations: 0-10000 nM Incubation Duration: 5 days Experimental Results: demonstrated strong inhibitory activities against these cells in 5-day assays. Western Blot Analysis[1] Cell Types: H526, A2780, ES-2, and MDA-MB231 cells Tested Concentrations: 0- 50 nM Incubation Duration: 24 hrs (hours) Experimental Results: Led to the depletion of phosphorylated BRD4 and c-MYC at 10 nM.
Animal Protocol	Animal/Disease Models: 4-6 weeks old female BALB/c nude mice/6-8 weeks old female C57BL/6 mice were used for B16F10 experiments (bearing NCI-H526, A2780, A375, B16F10, and TMD-8 cells)[1] Doses: 0.75-3 mg/kg Route of Administration: Po; TMD8 and B16F10 melanoma model with one time/day for 11-21 days; A375 melanoma and PDX of melanoma with one time/day (5 days on, 2 days off) for 21 days. Experimental Results: Strongly suppressed the growth of established lung tumor, ovarian tumor, lymphoma, and melanoma in vivo.
References	[1]. Potent BRD4 inhibitor suppresses cancer cell-macrophage interaction. Nat Commun. 2020;11(1):1833. Published 2020 Apr 14. [2]. Abstract 1382: A novel BET family bromodomain inhibitor NHWD-870 represents a promising therapeutic agent for a broad spectrum of cancers. Molecular and Cellular Biology, Genetics.

Solubility Data

Solubility (In Vitro)

DMSO : 62.5 mg/mL (127.14 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.23 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 20.8 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: ≥ 2.08 mg/mL (4.23 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 20.8 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	2.0342 mL	10.1711 mL	20.3422 mL
	5 mM	0.4068 mL	2.0342 mL	4.0684 mL
	10 mM	0.2034 mL	1.0171 mL	2.0342 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.