D-α-Hydroxyglutaric acid ((R)-2-Hydroxyglutarate) is the major metabolite accumulated in the neurometabolic disease D-2-hydroxyglutaric aciduria. D-α-Hydroxyglutaric acid is a weak competitive antagonist of α-ketoglutarate (α-KG), which can inhibit a variety of α-KG-dependent dioxygenases (dioxygenases) with a Ki of 10.87 mM. D-α-Hydroxyglutaric acid increases the production of reactive oxygen species (ROS). D-α-Hydroxyglutaric acid also binds to and inhibits ATP synthase and inhibits mTOR signaling.

Physicochemical Properties

Molecular Formula	C5H6O5-2
Molecular Weight	148.11
Exact Mass	148.037
CAS #	13095-47-1
PubChem CID	439391
Appearance	Typically exists as solid at room temperature
Melting Point	> 300 °C
LogP	-1
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	10
Complexity	141
Defined Atom Stereocenter Count	1
SMILES	O[C@@H](C(=O)O)CCC(=O)O
InChi Key	HWXBTNAVRSUOJR-GSVOUGTGSA-N
InChi Code	InChI=1S/C5H8O5/c6-3(5(9)10)1-2-4(7)8/h3,6H,1-2H2,(H,7,8)(H,9,10)/t3-/m1/s1
Chemical Name	(2R)-2-hydroxypentanedioic acid
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	Microbial Metabolite Human Endogenous Metabolite
ln Vitro	When isocitrate dehydrogenase (IDH) 1 and 2 genes undergo neomorphic mutations, D-α-hydroxyglutaric acid ((R)-2-hydroxyglutarate) builds up in human malignancies[1][2]. For both H3K9me2 and H3K27me2 peptides, 50 mM D-2-HG and 100 μM α-ketoglutarate (α-KG) cause a partial inhibition of KDM7A. The inhibition of Caenorhabditis elegans KDM7A (CeKDM7A) by 50 mM D-2-HG can be reversed by adding 300 μM α-KG, suggesting that D-2-HG is a weak competitive inhibitor against α-KG toward the CeKDM7A demethylase[1]. TET hydroxylases are weakly inhibited by D-α-hydroxyglutaric acid. Has a less noticeable inhibitory effect on TET1 when D-α-hydroxyglutaric acid is present at 0.1 mM[1].
ln Vivo	In rat cerebral cortex, human skeletal muscle, and submitochondrial particles from bovine heart, D-α-hydroxyglutaric acid significantly inhibits glucose utilization, CO2 generation, and the respiratory chain, indicating a dysfunction of the aerobic metabolism[5]. Because it dramatically reduced cell viability in neuronal cultures from chick embryo telencephalons and from neonatal rat hippocampus by stimulating certain NMDA glutamate receptors, hydroxyglutaric acid has also been suggested as an endogenous excitotoxic organic acid[5]. In the cortical supernatants of 30-day-old rats, D-α-Hydroxyglutaric acid (0.01 -1 mM) markedly boosted chemiluminescence and thiobarbituric acid-reactive substances (TBA-RS) while decreasing total antioxidant reactivity (TAR) values[5].
ADME/Pharmacokinetics	Metabolism / Metabolites 2-hydroxyglutarate can be converted to α-ketoglutaric acid through the action of a 2-hydroxyglutarate dehydrogenase.
Toxicity/Toxicokinetics	Toxicity Summary 2-hydroxyglutarate is an oncometabolite. It is a competitive inhibitor of multiple α-ketoglutarate-dependent dioxygenases, including histone demethylases and the TET family of 5-methlycytosine (5mC) hydroxylases. As a result, high levels of 2-hydroxyglutarate lead to genome-wide histone and DNA methylation alterations, which in turn lead to mutations that ultimately cause cancer. High L-2-Hydroxyglutarate levels are associated with gliomas and acute myeloid leukemia. L-2-Hydroxyglutarate can reversibly promote leukemogenesis in vitro. D-2-hydroxyglutarate mediates its neurotoxicity through activation of N-methyl-D-aspartate receptors. D-2-hydroxyglutarate is structurally similar to the excitatory amino acid glutamate and stimulates neurodegeneration by mechanisms well-known for glutamate, NMDA or mitochondrial toxins (A15198).
References	[1]. Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of α-ketoglutarate-dependent dioxygenases. Cancer Cell. 2011 Jan 18;19(1):17-30. [2]. 2-Hydroxyglutarate Inhibits ATP Synthase and mTOR Signaling. Cell Metab. 2015 Sep 1;22(3):508-15. [3]. Progress in understanding 2-hydroxyglutaric acidurias. J Inherit Metab Dis. 2012 Jul;35(4):571-87. [4]. D-2-hydroxyglutarate interferes with HIF-1α stability skewing T-cell metabolism towards oxidative phosphorylation and impairing Th17 polarization. Oncoimmunology. 2018 Mar 26;7(7):e1445454. [5]. D-2-hydroxyglutaric acid induces oxidative stress in cerebral cortex of young rats. Eur J Neurosci. 2003 May;17(10):2017-22.
Additional Infomation	(R)-2-hydroxyglutaric acid is the (R)-enantiomer of 2-hydroxyglutaric acid. It has a role as an algal metabolite, a human metabolite and a biomarker. It is a conjugate acid of a (R)-2-hydroxyglutarate(2-). It is an enantiomer of a (S)-2-hydroxyglutaric acid. D-2-Hydroxyglutaric acid is a metabolite found in or produced by Escherichia coli (strain K12, MG1655). (2R)-2-hydroxypentanedioic acid has been reported in Euphorbia resinifera, Chlamydomonas reinhardtii, and Trypanosoma brucei with data available. (R)-2-Hydroxyglutarate is the R-enantiomer of alpha-hydroxyglutarate (2-HG) where the hydroxy group is attached to the right side of the asymmetric carbon furthest from the carbonyl. This isoform is the predominant form found in gliomas associated with the expression of isocitrate dehydrogenase (IDH) mutants. Therefore, measurement of the expression of the R-form may be correlated more closely with those diseases than measurement of total 2-HG. D-2-Hydroxyglutaric acid is an alpha hydroxy acid. In humans the compound is formed by a hydroxyacid-oxoacid transhydrogenase whereas in bacteria is formed by a 2-hydroxyglutarate synthase. D-2-Hydroxyglutarate is also formed via the normal activity of hydroxyacid-oxoacid transhydrogenase during conversion of 4-hydroxybutyrate to succinate semialdehyde. The compound can be converted to α-ketoglutaric acid through the action of a 2-hydroxyglutarate dehydrogenase. In humans, there are two such enzymes called D2HGDH and L2HGDH. Tissue accumulation of high amounts of D-2-hydroxyglutaric acid is the biochemical hallmark of the inherited neurometabolic disorder D-2-hydroxyglutaric aciduria. Both the D and the L stereoisomers of hydroxyglutaric acid (EC 1.1.99.2) are found in body fluids. Accumulation of L-2-hydroxyglutaric acid has been reported in multiple patients who have a clinical phenotype of progressive neurodegeneration with extrapyramidal and cerebellar signs, seizures, and spongiform changes in the white matter. Patients who have excess accumulation of D-2-hydroxyglutaric acid in the urine exhibit a variable phenotype but included mental retardation, macrocephaly with cerebral atrophy, hypotonia, seizures, and involuntary movements. D-2-hydroxyglutarate mediates its neurotoxicity through activation of N-methyl-D-aspartate receptors. D-2-hydroxyglutarate is structurally similar to the excitatory amino acid glutamate and stimulates neurodegeneration by mechanisms well-known for glutamate, NMDA or mitochondrial toxins (A15198)

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	6.7517 mL	33.7587 mL	67.5174 mL
	5 mM	1.3503 mL	6.7517 mL	13.5035 mL
	10 mM	0.6752 mL	3.3759 mL	6.7517 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.