Artemisinin (also known as qinghaosu in Chinese; NSC-369397), discovered by Tu Youyou (2015 Nobel Prize winner in Medicine) team, is a naturally occurring anti-malarial drug isolated from the aerial parts of Artemisia annua L. plants. Artemisinin and its semi-synthetic derivatives are a group of drugs that possess the most rapid action of all current drugs against Plasmodium falciparum malaria.

Physicochemical Properties

Molecular Formula	C15H22O5
Molecular Weight	282.3322
Exact Mass	282.146
Elemental Analysis	C, 63.81; H, 7.85; O, 28.33
CAS #	63968-64-9
Related CAS #	Artemisinin-d3;176652-07-6
Appearance	White to off-white solid powder
Density	1.2±0.1 g/cm3
Boiling Point	389.9±42.0 °C at 760 mmHg
Melting Point	156-157ºC
Flash Point	172.0±27.9 °C
Vapour Pressure	0.0±0.9 mmHg at 25°C
Index of Refraction	1.533
LogP	2.27
SMILES	O1[C@@]23[C@]4([H])OC([C@]([H])(C([H])([H])[H])[C@]2([H])C([H])([H])C([H])([H])[C@@]([H])(C([H])([H])[H])[C@]3([H])C([H])([H])C([H])([H])C(C([H])([H])[H])(O1)O4)=O
InChi Key	BLUAFEHZUWYNDE-NNWCWBAJSA-N
InChi Code	InChI=1S/C15H22O5/c1-8-4-5-11-9(2)12(16)17-13-15(11)10(8)6-7-14(3,18-13)19-20-15/h8-11,13H,4-7H2,1-3H3/t8-,9-,10+,11+,13-,14-,15-/m1/s1
Chemical Name	(3R,5aS,6R,8aS,9R,12S,12aR)-3,6,9-trimethyloctahydro-12H-3,12-epoxy[1,2]dioxepino[4,3-i]isochromen-10(3H)-one
Synonyms	qinghaosu; NSC-369397; NSC369397; Arteannuin; Huanghuahaosu; Artemisinine; Artemisine; (+)-Artemisinin; NSC 369397
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

Targets	Plasmodium; anti-malarial
ln Vitro	Artemisinin (3.125-100 μM) concentration-dependently suppresses Aβ25-35 induced cytotoxicity in PC12 cells. Artemisinin (25 μM) suppresses Aβ25-35-induced LDH release, apoptosis and ROS production, attenuates Aβ-induced mitochondrial membrane potential loss and caspase 3/7 activity increase, and stimulates the phosphorylation of ERK1/2 in a time- and concentration-dependent manner in PC12 cell. ERK 1/2 pathway mediates the protect effects of artemisinin in PC12 cells[1]. Artemisinin shows cytotoxic activity in MCF-7/Dox cell line with IC50 of 3.7±0.4 μg/mL after 24 h treatment. Besides, Artemisinin treatment of MCF-7 cells, sensitive and resistant to Dox and DDP, causes a decrease in expression of proteins such as LF, FTH1, and HEP. Artemisinin activates p38 MAPK-kinase cascade regardless of the oxidative stress due to inhibition of VEGF expression and cell migration[2]. Artemisinin (50, 100 or 200 mg) significantly inhibits isoflurane-induced hippocampal neuronal loss, decreases caspase-3-positive cell counts and also cleaves caspase-3 expression, and modulates the expression of apoptosis pathway proteins. Artemisinin modulates JNK/ERK 1/2 signalling and histone acetylation[3]. Artemisinin inhibits HCV replication in a dose-dependent manner with EC50 value of 167±38 µM. Artemisinin and its most potent analogues partially inhibit the in vitro replication of HCV by induction of reactive oxygen species (ROS)[4]. Artemisinin significantly inhibits VSMC proliferation in a dose-dependent manner. Artemisinin (1 mM) for 72 h significantly reduces the expression of proliferating cell nuclear antigen messenger RNA[5].
ln Vivo	Artemisinin (50, 100 or 200 mg/kg b.wt/day, p.o.) prevents isoflurane-induced working memory impairments as observed in T-maze test. Artemisinin enhances spatial navigation and memory of rats exposed to isoflurane. Artemisinin-treated rats exhibit markedly better performance in comparison with isoflurane-alone-exposed rats[3].
Enzyme Assay	Reactive Oxygen Species Measurements[2] Intracellular and mitochondrial ROS production were assessed by a fluorometric assay using 2′,7′-dichlorofluorescein diacetate (H2DCFDA) and MitoTracker Red CMXRos, respectively. After 12 h treatment of artemisinin and followed by 12 h treatment of glutamate, cells were incubated in 10 μmol/L H2DCFDA or 0.25 μmol/L MitoTracker Red CMXRos for 30 min at 37°C. The fluorescence was then observed via a fluorescence microscope. The fluorescence was detected with 530/485-nm and 579/599-nm excitation/emission wave lengths.[2] Mitochondrial Membrane Potential (ΔΨm) Measurement[2] The mitochondrial membrane potential was detected using Tetramethylrhodamine, Ethyl Ester (TMRE) mitochondrial membrane potential assay kit (Abcam, USA). Cells were loaded with 20 nM of TMRE working solution for 20 min at 37°C. The fluorescent images were observed and obtained on a Zeiss fluorescence microscope. Fluorescence intensity was measured using a Tecan Infinite F200 plate reader with 594/575-nm excitation/emission.
Cell Assay	For this purpose, cells are cultivated in 96-well plates in DMEM, supplemented with insulin. The artemisinin, Dox, and DDP are added to media at different concentrations and the cells are cultivated for either 24 or 48 h. For this purpose artemisinin is diluted in 0.01% DMSO in media. After this time, 10 µL of the MTT dye solution (5 mg/mL in phosphate buffer saline) is added to the cells; the cells are incubated at the same conditions for 3 h. After centrifugation (1500 rpm, 5 min) the supernatant is removed. 100 μL of dimethyl sulfoxide is added to each well, to dissolve formazan. The absorption is measured, using a multi-well spectrophotometer at a wavelength of 540 nm.
Animal Protocol	Separate group of rat pups (total rat pups 80; n = 16 per group) is administered artemisinin (50, 100 or 200 mg/kg body weight) via oral gavage, every day from P2 to P21. On P7, the pups are exposed to isoflurane (0.75% in 30% oxygen or air) for 6 h in a temperature-controlled chamber. Animals that are not exposed to anaesthesia nor given artemisinin served as control group, while rats that receive isoflurane alone are grouped as anaesthetic-controls.
References	[1]. Artemisinin protects PC12 cells against β-amyloid-induced apoptosis through activation of the ERK1/2 signaling pathway. Redox Biol. 2017 Apr 4;12:625-633. [2]. Artemisinin Prevents Glutamate-Induced Neuronal Cell Death Via Akt Pathway Activation. Front Cell Neurosci. 2018 Apr 20;12:108.
Additional Infomation	(+)-artemisinin is a sesquiterpene lactone obtained from sweet wormwood, Artemisia annua, which is used as an antimalarial for the treatment of multi-drug resistant strains of falciparum malaria. It has a role as an antimalarial and a plant metabolite. It is a sesquiterpene lactone and an organic peroxide. Artemisinin has been used in trials studying the treatment of Schizophrenia, Malaria, Falciparum, and Plasmodium Falciparum. Artemisinin has been reported in Artemisia lancea, Artemisia annua, and other organisms with data available. Accumulating evidence displays that an abnormal deposition of amyloid beta-peptide (Aβ) is the primary cause of the pathogenesis of Alzheimer's disease (AD). And therefore the elimination of Aβ is regarded as an important strategy for AD treatment. The discovery of drug candidates using culture neuronal cells against Aβ peptide toxicity is believed to be an effective approach to develop drug for the treatment of AD patients. We have previously showed that artemisinin, a FDA-approved anti-malaria drug, has neuroprotective effects recently. In the present study, we aimed to investigate the effects and potential mechanism of artemisinin in protecting neuronal PC12 cells from toxicity of β amyloid peptide. Our studies revealed that artemisinin, in clinical relevant concentration, protected and rescued PC12 cells from Aβ25-35-induced cell death. Further study showed that artemisinin significantly ameliorated cell death due to Aβ25-35 insult by restoring abnormal changes in nuclear morphology, lactate dehydrogenase, intracellular ROS, mitochondrial membrane potential and activity of apoptotic caspase. Western blotting analysis demonstrated that artemisinin activated extracellular regulated kinase ERK1/2 but not Akt survival signaling. Consistent with the role of ERK1/2, preincubation of cells with ERK1/2 pathway inhibitor PD98059 blocked the effect of artemisinin while PI3K inhibitor LY294002 has no effect. Moreover, Aβ1-42 also caused cells death of PC12 cells while artemisinin suppressed Aβ1-42 cytotoxicity in PC12 cells. Taken together, these results, at the first time, suggest that artemisinin is a potential protectant against β amyloid insult through activation of the ERK1/2 pathway. Our finding provides a potential application of artemisinin in prevention and treatment of AD.[1] Artemisinin is an anti-malarial drug that has been in use for almost half century. Recently, novel biological effects of artemisinin on cancer, inflammation-related disorders and cardiovascular disease were reported. However, neuroprotective actions of artemisinin against glutamate-induced oxidative stress have not been investigated. In the current study, we determined the effect of artemisinin against oxidative insult in HT-22 mouse hippocampal cell line. We found that pretreatment of artemisinin declined reactive oxygen species (ROS) production, attenuated the collapse of mitochondrial membrane potential induced by glutamate and rescued HT-22 cells from glutamate-induced cell death. Furthermore, our study demonstrated that artemisinin activated Akt/Bcl-2 signaling and that neuroprotective effect of artemisinin was blocked by Akt-specific inhibitor, MK2206. Taken together, our study indicated that artemisinin prevented neuronal HT-22 cell from glutamate-induced oxidative injury by activation of Akt signaling pathway.[2]

Solubility Data

Solubility (In Vitro)

DMSO : 50~56 mg/mL (177.10~198.34 mM) H2O : < 0.1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (7.37 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 (7.37 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: ≥ 2.08 mg/mL (7.37 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 3% DMSO+ 97% Corn oil: 6mg/ml (21.25mM)  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.5420 mL	17.7098 mL	35.4195 mL
	5 mM	0.7084 mL	3.5420 mL	7.0839 mL
	10 mM	0.3542 mL	1.7710 mL	3.5420 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.