Anti-inflammatory agent 35 (compound 5a27) is an orally bioavailable curcumin analog with anti-inflammatory activity. Anti-inflammatory agent 35 blocks mitogen-activated protein kinase (MAPK) signaling and nuclear translocation of NF-kB. Anti-inflammatory agent 35 also inhibits yellow neutrophil infiltration and proinflammatory cytokine production. Anti-inflammatory agent 35 significantly attenuated lipopolysaccharide (LPS)-induced acute lung injury (ALI) in in vivo studies.

Physicochemical Properties

Molecular Formula	C27H29NO8
Molecular Weight	495.52
Exact Mass	495.189
CAS #	2293951-00-3
PubChem CID	155549737
Appearance	White to off-white solid powder
LogP	3.7
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	9
Heavy Atom Count	36
Complexity	826
Defined Atom Stereocenter Count	0
SMILES	N1(C(=O)/C=C/C2=CC=C(OC(C)=O)C(OC)=C2)CCC/C(=C\C2=CC(OC)=C(OC)C(OC)=C2)/C1=O
InChi Key	QLGWATTXCFBRGY-ZEQSIELKSA-N
InChi Code	InChI=1S/C27H29NO8/c1-17(29)36-21-10-8-18(14-22(21)32-2)9-11-25(30)28-12-6-7-20(27(28)31)13-19-15-23(33-3)26(35-5)24(16-19)34-4/h8-11,13-16H,6-7,12H2,1-5H3/b11-9+,20-13+
Chemical Name	[2-methoxy-4-[(E)-3-oxo-3-[(3E)-2-oxo-3-[(3,4,5-trimethoxyphenyl)methylidene]piperidin-1-yl]prop-1-enyl]phenyl] acetate
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

Targets	p38 MAPK ERK I-kappaBalpha IL-6 p65 NF-κB
ln Vitro	In mice, the development of pro-inflammatory cytokines (IL-6, TNF-α) by LPS (0.5 μg/mL; 24 hours) is inhibited by anti-inflammatory drug 35 (compound 5a27) (10 μM; 30 minutes). It is not cytotoxic to produce macrophages. With an IC50 of 2.40 μM and 2.23 μM, respectively, anti-inflammatory drug 35 suppresses the synthesis of TNF-α and IL-6 [1]. RAW 264.7 murine macrophages' activation caused by LPS was greatly reduced by anti-inflammatory drug 35 (10 μM; 30 min). Furthermore, it suppresses IκB levels, p-p38, and p-ERK substantially, which suggests that MAPK and NF-κB signaling are inhibited [1].
ln Vivo	Curcumin (HY-N0005) had a lower bioavailability than anti-inflammatory drug 35 (Compound 5a27) (50 mg/kg; oral; single dose, measured 0–25 minutes) [1]. By reducing inflammation in mice models, anti-inflammatory drug 35 (10 mg/kg; intraperitoneal injection; once daily for 1 week) ameliorates LPS-induced ALI [1]. Rat pharmacokinetic parameters [1]: Route Dose (mg/kg) AUC(0-t) (μg/L·h) AUC(0-∞) (μg/L·h) MRT(0-) (h) t1/2 (h) Tmax (h) CLz/F (L/h/kg) Vz/F (L/kg) Cmax po 50 231.2 325.6 7.8 12.3 6.7 3.3 5062.6 827.1 113.3 iv 5 34.3 122.4 11.3 19.9 0.2 0.1 404.4 59.5 16.4
Cell Assay	Western Blot Analysis[1] Cell Types: RAW 264.7 mouse macrophages Tested Concentrations: 10 μM Incubation Duration: 30 min; followed by incubation with 0.5 μg/mL LPS for another 30 min Experimental Results: diminished the phosphorylation of p38 and ERK. And down-regulated IκB (inhibitor of NF-κB), inhibits the transcription of TNF-α, IL-6, IL-1β, ICAM-1.
Animal Protocol	Animal/Disease Models: ALI mouse model (C57BL/6 mice)[1] Doses: 10 mg/kg Route of Administration: intraperitoneal (ip)injection, one time/day for 1 week; 30 min later every dose, followed by 5 mg/kg LPS, intratracheal injection Experimental Results: Dramatically normalized the wet/dry ratio of lungs.
References	[1]. Design and synthesis novel di-carbonyl analogs of curcumin (DACs) act as potent anti-inflammatory agents against LPS-induced acute lung injury (ALI). Eur J Med Chem. 2019 Apr 1;167:414-425.

Solubility Data

Solubility (In Vitro)

DMSO : 16.67 mg/mL (33.64 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 1.67 mg/mL (3.37 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 16.7 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	2.0181 mL	10.0904 mL	20.1808 mL
	5 mM	0.4036 mL	2.0181 mL	4.0362 mL
	10 mM	0.2018 mL	1.0090 mL	2.0181 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.