ALX-1393 TFA is a selective GlyT2 inhibitor (antagonist) with antinociceptive effects on thermal, mechanical and chemical stimuli in a rat model of acute pain.

Physicochemical Properties

Molecular Formula	C25H23F4NO6
Molecular Weight	509.45
Related CAS #	ALX-1393;949164-09-4
PubChem CID	168475830
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	11
Rotatable Bond Count	9
Heavy Atom Count	36
Complexity	583
Defined Atom Stereocenter Count	1
SMILES	C1=CC=C(C=C1)COC2=CC=CC=C2C(C3=CC(=CC=C3)F)OC[C@@H](C(=O)O)N.C(=O)(C(F)(F)F)O
InChi Key	UDSOPFWCTVYJOU-CNAJMHLNSA-N
InChi Code	InChI=1S/C23H22FNO4.C2HF3O2/c24-18-10-6-9-17(13-18)22(29-15-20(25)23(26)27)19-11-4-5-12-21(19)28-14-16-7-2-1-3-8-16;3-2(4,5)1(6)7/h1-13,20,22H,14-15,25H2,(H,26,27);(H,6,7)/t20-,22?;/m0./s1
Chemical Name	(2S)-2-amino-3-[(3-fluorophenyl)-(2-phenylmethoxyphenyl)methoxy]propanoic acid;2,2,2-trifluoroacetic acid
Synonyms	ALX-1393 (TFA); ALX-1393 TFA; ALX1393 (TFA); ALX 1393 TFA;
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, 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	Glycine Transporter 2 (GlyT2) [1][2]
ln Vivo	Intrathecal administration of ALX1393 (10–30 nmol) in rats significantly increased paw withdrawal thresholds to mechanical stimuli (von Frey test) and reduced formalin-induced Phase 2 flinching behavior, indicating potent antinociception in acute pain models. This effect was blocked by the glycine receptor antagonist strychnine. [1] Intracerebroventricular (i.c.v.) injection of ALX1393 (0.1–10 μg) in rats attenuated thermal hyperalgesia in the Complete Freund's Adjuvant (CFA)-induced inflammatory pain model and mechanical allodynia in the sciatic nerve ligation-induced neuropathic pain model. Efficacy was stronger in inflammatory pain than neuropathic pain. [2] In normal rats, ALX1393 TFA (icv; 25, 50, and 100 μg) decreases the late-phase response in the formalin test without affecting motor function. In a dose-dependent manner, ALX1393 suppresses both mechanical and cold hyperalgesia[2].
Animal Protocol	In normal rats, ALX1393 TFA (icv; 25, 50, and 100 μg) decreases the late-phase response in the formalin test without affecting motor function. In a dose-dependent manner, ALX1393 suppresses both mechanical and cold hyperalgesia[2].
References	[1]. The antinociceptive effect of intrathecal administration of glycine transporter-2 inhibitor ALX1393 in a rat acute pain model. Anesth Analg. 2010 Feb 1;110(2):615-21. [2]. Antinociceptive effect of intracerebroventricular administration of glycine transporter-2 inhibitor ALX1393 in rat models of inflammatory and neuropathic pain. Pharmacol Biochem Behav. 2015 Mar;130:46-52.
Additional Infomation	ALX1393 exerts antinociception by inhibiting GlyT2 in the spinal cord, thereby increasing synaptic glycine levels and enhancing glycinergic inhibitory neurotransmission. This mechanism is validated by strychnine reversibility. [1] The differential efficacy of i.c.v.-administered ALX1393 in inflammatory vs. neuropathic pain suggests region-specific GlyT2 modulation in supraspinal pain pathways. Notably, it did not impair motor function (rotarod test). [2] CNS-penetrant GlyT2 inhibitor showing superior efficacy in neuropathic vs inflammatory pain models [2].

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	1.9629 mL	9.8145 mL	19.6290 mL
	5 mM	0.3926 mL	1.9629 mL	3.9258 mL
	10 mM	0.1963 mL	0.9815 mL	1.9629 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.