Oxidopamine (6-OHDA) HBr is an antagonist of the neurotransmitter dopamine. Oxidopamine HBr is a extensively used neurotoxin that selectively destroys dopaminergic neurons. Oxidopamine HBr promotes COX-2 activation, leading to PGE2 synthesis and secretion of the pro-inflammatory cytokine IL-1β. Oxidopamine HBr may be utilized in study/research of Parkinson's disease (PD), attention deficit hyperactivity disorder (ADHD), and Leschnain syndrome.

Physicochemical Properties

Molecular Formula	C8H12BRNO3
Molecular Weight	250.09
Exact Mass	249
Elemental Analysis	C, 38.42; H, 4.84; Br, 31.95; N, 5.60; O, 19.19
CAS #	636-00-0
Related CAS #	Oxidopamine hydrochloride;28094-15-7; 1199-18-4; 636-00-0 (HBr)
PubChem CID	176170
Appearance	Light brown to gray solid
Boiling Point	406ºC at 760 mmHg
Melting Point	216-220 °C(lit.)
Flash Point	199.3ºC
LogP	1.963
Hydrogen Bond Donor Count	5
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	2
Heavy Atom Count	13
Complexity	142
Defined Atom Stereocenter Count	0
SMILES	OC1=CC(CCN)=C(O)C=C1O.[H]Br
InChi Key	MLACDGUOKDOLGC-UHFFFAOYSA-N
InChi Code	InChI=1S/C8H11NO3.BrH/c9-2-1-5-3-7(11)8(12)4-6(5)10;/h3-4,10-12H,1-2,9H2;1H
Chemical Name	2,4,5-Trihydroxyphenethylamine hydrobromide
Synonyms	6-Hydroxydopamine hydrobromide; 6-OHDA hydrobromide; 6-OHDA HBr; 6 OHDA HBr; 6OHDA HBr; 6-OHDA Hydrobromide; 6 OHDA Hydrobromide; 6OHDA Hydrobromide; 6-Hydroxydopamine hydrobromide; 636-00-0; Oxidopamine hydrobromide; Oxidopamine (hydrobromide); 6-hydroxydopamine hbr; 6-OHDA; 2,4,5-Trihydroxyphenethylamine hydrobromide; 5-(2-aminoethyl)benzene-1,2,4-triol hydrobromide; 6-Hydroxydopamine Hydrobromide; 6Hydroxydopamine Hydrobromide; 6 Hydroxydopamine Hydrobromide; 6-Hydroxydopamine HBr; 6 Hydroxydopamine HBr
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: (1). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.(2). This product is not stable in solution, please use freshly prepared working solution for optimal results.
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	COX-2; IL-1;βCaspase-3; Caspase-8; Caspase-9
ln Vitro	Dopamine oxidation by hydrobromic acid (0-500 μM, 24 hours) decreases Neuro-2a cell survival and SH-SY5Y cell viability in a concentration-dependent way [1]. Dopamine oxidation with hydrobromic acid (75-150 μM, 0-24 h) stimulates the expression of COX-2. It also promotes the production of pro-inflammatory cytokines such IL-1β and PGE 2 biosynthesis [1]. Dopamine (0-150 μM, 12 h) and nuclear translocation are oxidized by hydrobromic acid [1]. Phosphorylation of p38 is induced by hydrobromic acid oxidation of dopamine (75 μM, 0–12 h) [3].
ln Vivo	Degeneration of dopaminergic neurons in the substantia nigra is caused by hydrobromic acid oxidation of dopamine (5 μg/2 μL, injected unilaterally into the right striatum) [2].
Enzyme Assay	Cyclooxygenase-2 (COX-2) triggers pro-inflammatory processes that can aggravate neuronal degeneration and functional impairments in many neurological conditions, mainly via producing prostaglandin E2 (PGE2) that activates four membrane receptors, EP1-EP4. However, which EP receptor is the culprit of COX-2/PGE2-mediated neuronal inflammation and degeneration remains largely unclear and presumably depends on the insult types and responding components. Herein, we demonstrated that COX-2 was induced and showed nuclear translocation in two neuronal cell lines - mouse Neuro-2a and human SH-SY5Y - after treatment with neurotoxin 6-hydroxydopamine (6-OHDA), leading to the biosynthesis of PGE2 and upregulation of pro-inflammatory cytokine interleukin-1β. Inhibiting COX-2 or microsomal prostaglandin E synthase-1 suppressed the 6-OHDA-triggered PGE2 production in these cells. Treatment with PGE2 or EP2 selective agonist butaprost, but not EP4 agonist CAY10598, increased cAMP response in both cell lines. PGE2-initiated cAMP production in these cells was blocked by our recently developed novel selective EP2 antagonists - TG4-155 and TG6-10-1, but not by EP4 selective antagonist GW627368X. The 6-OHDA-promoted cytotoxicity was largely blocked by TG4-155, TG6-10-1 or COX-2 selective inhibitor celecoxib, but not by GW627368X. Our results suggest that PGE2 receptor EP2 is a key mediator of COX-2 activity-initiated cAMP signaling in Neuro-2a and SH-SY5Y cells following 6-OHDA treatment, and contributes to oxidopamine-mediated neurotoxicity [1].
Cell Assay	Cell Viability Assay[1] Cell Types: Neuro-2a cells and SH-SY5Y cells Tested Concentrations: 0-500 µM Incubation Duration: 24 or 48 hrs (hours) Experimental Results: Induction of neurotoxicity in Neuro-2a cells and SH-SY5Y cells in a concentration-dependent manner Causes cytotoxicity in SY5Y cells. In Neuro-2a cells, EC50=111 µM (incubation for 24 hrs (hours)) and 109 µM (incubation for 48 hrs (hours)); in SH-SY5Y cells, EC50=118 µM for 24 hrs (hours) and EC50=107 µM for 48 hrs (hours). RT-PCR[1] Cell Types: Neuro-2a cells and SH-SY5Y cells Tested Concentrations: 75 or 150 µM Incubation Duration: 0, 6 or 24 hrs (hours) Experimental Results: Rapid and robust induction of COX-2 in a time-dependent manner. Induces COX-2 activation, characterized by induction of expression and nuclear translocation. PGE2 in the culture medium increased Dramatically by nearly 5-fold in Neuro-2a cells (75 µM) and 3-fold in SH-SY5Y cells (150 µM). The pro-inflammatory cytokine interleukin 1β (IL-1β) was Dramatically upregulated in Neuro-2a cells and SH-SY5Y cells. Apoptosis analysis [3] Cell Types: PC12 cells Tested Concentrations: 0, 25, 50, 75, and 150 μM Incubation Duration: 0, 2, 4, 6, 12, and 20 h Experimental Results: Induced apoptosis of PC12 cells. Increased the activities of caspase-3, -8 and -9 in PC12 cells in a time- and concentration-dependent manner. Increased these caspase activities at 2-4 h and reached a maximum at 12 h. diminished cells with high mitochondrial membrane potential (JC-1 aggregate) in a time- and concentration-dependent manner. Western Blot Analysis[3] Cell Types: PC12 cells Tested Concentrations: 75 μM Incubation Duration: 0, 3, 5, 6, 8, 10, and 12 h Experimental Results: Increased the level of p-p38 in a time-dependent manner.
Animal Protocol	The present study was undertaken to investigate the neuroprotective effects of resveratrol on 6-hydroxydopamine (6-OHDA)-induced Parkinson's disease in rats. 6-OHDA-induced Parkinson's disease rat model involves chronic inflammation, mitochondrial dysfunction, and oxidative stress, and the loss of the dopaminergic neurons in the substantia nigra is the predominant lesion. Resveratrol has been shown to have anti-inflammatory actions, and thus was tested for its beneficial effects using 6-OHDA-induced Parkinson's disease rat model. Adult Sprague-Dawley (SD) rats were unilaterally injected with 6-OHDA (5 microg/2 microl) into the right striatum, and the striatum damage was assessed by rotational test, ultrahistopathology, and molecular alterations. Resveratrol (10, 20 and 40 mg/kg) was then given orally to Parkinson's disease rats, daily for 10 weeks to examine the protective effects. Rotational test (turns of rats) showed that resveratrol significantly attenuated apomorphine-induced turns of rats in 6-OHDA-injuried Parkinson's disease rat model as early as two weeks of administration. Ultrastructural analysis showed that resveratrol alleviated 6-OHDA-induced chromatin condensation, mitochondrial tumefaction and vacuolization of dopaminergic neurons in rat substantia nigra. Furthermore, resveratrol treatment also significantly decreased the levels of COX-2 and TNF-alpha mRNA in the substantia nigra as detected by real-time RT-PCR. COX-2 protein expression in the substantia nigra was also decreased as evidenced by Western blotting. These results demonstrate that resveratrol exerts a neuroprotective effect on 6-OHDA-induced Parkinson's disease rat model, and this protection is related to the reduced inflammatory reaction.[2].
References	[1]. Cyclooxygenase-2 contributes to oxidopamine-mediated neuronal inflammation and injury via the prostaglandin E2 receptor EP2 subtype. Sci Rep. 2017 Aug 25;7(1):9459. [2]. Neuroprotective effect of resveratrol on 6-OHDA-induced Parkinson's disease in rats. Eur J Pharmacol. 2008 Dec 14;600(1-3):78-82. [3]. Cell-permeable cAMP analog suppresses 6-hydroxydopamine-induced apoptosis in PC12 cells through the activation of the Akt pathway. Brain Res. 2006 Oct 3;1113(1):10-23. [4]. Autoxidation and neurotoxicity of 6-hydroxydopamine in the presence of some antioxidants: potential implication in relation to the pathogenesis of Parkinson's disease. J Neurochem. 2000 Apr;74(4):1605-12.
Additional Infomation	A neurotransmitter analogue that depletes noradrenergic stores in nerve endings and induces a reduction of dopamine levels in the brain. Its mechanism of action is related to the production of cytolytic free-radicals.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~199.93 mM) H2O : ~20 mg/mL (~79.97 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (10.00 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 25.0 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 (8.32 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 20.8 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: 50 mg/mL (199.93 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.9986 mL	19.9928 mL	39.9856 mL
	5 mM	0.7997 mL	3.9986 mL	7.9971 mL
	10 mM	0.3999 mL	1.9993 mL	3.9986 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.