TML-6 is an orally bioavailable curcumin analogue that can inhibit the synthesis of β-amyloid precursor protein and β-amyloid (Aβ). TML-6 upregulates Apo E, inhibits NF-κB and mTOR, and increases the activity of the antioxidant Nrf2 gene. TML-6 may be utilized in AD/Alzheimer's disease research.

Physicochemical Properties

Molecular Formula	C30H37NO7
Molecular Weight	523.617289304733
Exact Mass	523.257
CAS #	1462868-88-7
Related CAS #	TML-6-d3;2673270-28-3
PubChem CID	72705569
Appearance	Off-white to yellow solid powder
LogP	4.4
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	14
Heavy Atom Count	38
Complexity	783
Defined Atom Stereocenter Count	0
SMILES	O=C(CC(C(/C=C/C1C=CC(=C(C=1)OC)OC)=O)(C(/C=C/C1C=CC(=C(C=1)OC)OC)=O)C)N(CC)CC
InChi Key	UWVCYNXVZRDWSD-UNZYHPAISA-N
InChi Code	InChI=1S/C30H37NO7/c1-8-31(9-2)29(34)20-30(3,27(32)16-12-21-10-14-23(35-4)25(18-21)37-6)28(33)17-13-22-11-15-24(36-5)26(19-22)38-7/h10-19H,8-9,20H2,1-7H3/b16-12+,17-13+
Chemical Name	(E)-6-(3,4-dimethoxyphenyl)-3-[(E)-3-(3,4-dimethoxyphenyl)prop-2-enoyl]-N,N-diethyl-3-methyl-4-oxohex-5-enamide
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: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.
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	NF-κB mTOR
ln Vitro	TML-6 (0.65–5.24 µg/mL; for 24 hours) increases the expression of ApoE protein while decreasing that of APP and phospho-NF-κB. TML-6 suppresses phospho-mTOR, which in turn inhibits the mTOR signaling pathway[1]. With an IC50 of 4.19 µg/mL (8 μM), TML-6 (0.31, 0.63, 2.5, 5, 10, 20 μM; 24 h) exhibits minimal cytotoxicity in Huh-7 cells at concentrations below 5 μM[1]. In N2a/APPswe cells, TML-6 (1.05, 2.09, 3.14, 4.19 μg/mL; 24 h) dose-dependently decreases the production of Aβ40 and Aβ42 between 1.05, 2.09, and 3.14 μg/mL (equivalent to 2, 4 and 6 μM)[1]. TML-6 has the ability to activate the Nrf2 gene transcriptionally in a dose-dependent manner; the maximum activity is observed at 1.32 µg/mL[1].
ln Vivo	Treatment with TML-6 (diet; 150 mg/kg/day; for four months) significantly improves learning and reduces Aβ in the brain and the microglial activation marker Iba-1[1]. The T1/2, Cmax, and AUC of TML-6 (oral; 150 mg/kg) are 1.27 hours, 35.9 ng/mL, and 177 ng·hr/mL, respectively[1].
Cell Assay	Western Blot Analysis[1] Cell Types: Huh-7 cells Tested Concentrations: 0.65, 1.31, 1.96, 2.61, 3.93, 5.24 µg/mL Incubation Duration:For 24 hrs (hours) Experimental Results: diminished the amyloid precursor protein (APP) protein expression level by 60 % and diminished the level of phosphorylated NF-κB by about 50% at a dose of 1.96 µg/mL after 24 h treatment. Induced the protein expression level of ApoE by approximately 44% at a dose of 2.62 µg/mL.
Animal Protocol	Animal/Disease Models: Six-month-old 3xTg (mutations: APPKM670 /671NL, MAPTP301L and PSEN1M146V) AD transgenic mice[1] Doses: 150 mg/kg Route of Administration: Diet; daily; for four months Experimental Results: Improved the learning behaviors, Dramatically suppressed the Aβ levels and Iba-1 expression in the brain of 3xTg AD transgenic mice. Animal/Disease Models: SD rats[1] Doses: 150 mg/kg (pharmacokinetic/PK Analysis) Route of Administration: Oral Experimental Results: Had a T1/2 of 1.27 hrs (hours), a Cmax of 35.9 ng/mL and an AUC of 177 ng• hr/mL.
References	[1]. A Curcumin Analog Exhibits Multiple Biologic Effects on the Pathogenesis of Alzheimer's Disease and Improves Behavior, Inflammation, and β-Amyloid Accumulation in a Mouse Model. Int J Mol Sci. 2020 Jul 30;21(15):5459.

Solubility Data

Solubility (In Vitro)

DMSO: 120 mg/mL (229.17 mM)

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	1.9098 mL	9.5489 mL	19.0978 mL
	5 mM	0.3820 mL	1.9098 mL	3.8196 mL
	10 mM	0.1910 mL	0.9549 mL	1.9098 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.