(R,R)-BD-AcAc 2 (D-3HHB) enhances blood Na+ and creatinine levels, decreases Aldh3b2 gene expression, increases blood glucose concentration moderately, boosts particular muscle strength, and lowers plasma free fatty acid concentration [1].

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Racemic 1,3-butanediol acetoacetate diester was shown to increase ketone body concentrations in the blood of both a pig and a dog. [2]
More recent studies in rats indicate it induces a rapid increase in (R)-β-hydroxybutyric acid concentration in the blood and a decrease in blood glucose concentration, indicative of ketosis. [2]
(±)-1,3-Butanediol acetoacetate diester has been shown to suppress seizure activity in rats. [2]
Combining (±)-1,3-butanediol acetoacetate diester with hyperbaric oxygen therapy was shown to slow metastatic cancer growth in mice. [2]

Animal/Disease Models: Septic mice [1]
Doses: 10, 20, 40 and 80 mmol/kg/day
Route of Administration: PO
Experimental Results: Specific muscle strength increased to 40 mmol/kg/day compared to placebo Healthy 93% of control levels. There was a modest increase in blood glucose concentrations of 40 mmol/kg/day compared to placebo. A dose of 40 mmol/kg/day diminished Aldh3b2 gene expression compared with placebo. Caused a further moderate increase in blood Na+ levels and an increase in blood creatinine levels to 20 mmol/kg/day. Plasma free fatty acid concentrations decrease by 10 or 20 mmol/kg/day. Liver gene expression levels of Aldh1a7 were also diminished by sepsis but increased by D-3HHB.

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28-Day Oral Toxicity Study: Male and female CrI:WI (Wistar) rats (approximately 9 weeks old) were used. The test group received a diet formulated to contain 30% of calories from (R)-3-hydroxybutyl (R)-3-hydroxybutyrate (ketone monoester), which resulted in daily intakes of 12.0 g/kg body weight/day for males and 15.1 g/kg body weight/day for females. The diet was prepared by mixing rodent chow powder with sugar-free jelly crystals, then adding a heated mixture of water and the ketone monoester to form pellets. Control groups received isocaloric diets where the ketone monoester calories were replaced with either carbohydrate (corn starch) or fat (palm oil). All diets were matched for protein content. Animals were fed ad libitum for 28 days. Body weights and food consumption were recorded regularly. At termination (day 29), blood was collected for hematology, coagulation, and clinical chemistry analysis. Urine was collected for urinalysis. A full gross necropsy was performed, and organs were weighed. Liver, kidneys, gastrointestinal tract sections, brain, heart, and skeletal muscle were preserved for histopathological examination. [3]
Developmental Toxicity Study: Pregnant CrI:WI (Han) rats were administered (R)-3-hydroxybutyl (R)-3-hydroxybutyrate (neat, undiluted) at a dosage of 2000 mg/kg body weight/day (2 g/kg/day) via oral gavage from gestation days (DG) 6 through 20. The control group received an equivalent volume of reverse osmosis (RO) water. Dams were observed for clinical signs, and body weights and food consumption were recorded. On DG 21, dams were euthanized, subjected to Caesarean section, and examined. Maternal blood was collected for hematology and clinical chemistry. The uterus was examined for implantation sites, resorptions, and live fetuses. Fetuses were examined for external, visceral, and skeletal alterations. [3]

(±)-1,3-Butanediol acetoacetate diester is metabolically hydrolyzed to produce two equivalents of acetoacetic acid and (±)-1,3-butanediol. [2] (±)-1,3-Butanediol can be oxidized to β-hydroxybutyric acid. Oxidation of the (R)-enantiomer occurs in the liver. (S)-β-hydroxybutyric acid is not naturally occurring, but its metabolism in perfused rat liver has been shown to produce ketone bodies via an unknown pathway. [2]

28-day study showed that rats fed a ketomonose diet consumed less food and gained significantly less weight than control rats fed a carbohydrate or fat diet. Hematological examinations revealed significantly higher red blood cell counts, hemoglobin, and hematocrit in both male and female rats in the ketomonose diet compared to the control group; male rats also showed higher reticulocyte counts. All parameters were within the normal physiological range. The activated partial thromboplastin time (APTT) was slightly shorter in the ketomonose diet, but still within the normal range. Clinical chemistry examinations showed significantly higher levels of creatine kinase (CK), albumin, and alanine aminotransferase (ALT) in male rats compared to the control group, and elevated levels of lactate dehydrogenase (LDH) in both male and female rats. Most values were within the normal range, but LDH levels were slightly above the historical limit in some cases. Cholesterol levels were significantly higher in the ketomonose diet compared to the control group. Urinalysis revealed no treatment-related abnormalities.
The absolute uterine weight of female rats in the ketone ester feeding group was lower, but this was attributed to their lower body weight, and the relative uterine weight was comparable to that of the control group.
Histopathological examination showed vacuolation (lipid accumulation) in the liver of female rats in all diet groups (experimental and control groups) and a few male control rats. Mild necrotic inflammatory foci in the liver and mild myocyte necrosis/focal histiocytosis in skeletal muscle were observed in some animals in all groups. These findings were considered to be due to background factors or dietary dilution and were not specific to ketone monoester. No other toxicologically significant histopathological lesions were found associated with the test substance. [3] Developmental toxicity studies: Female rats given 2 g/kg ketone monoester daily showed reduced weight gain (adjusted for gestational uterine weight) and reduced food intake compared to the control group. Hematological examination of the female rats showed increased lobar neutrophils and decreased basophils in the experimental group, which was considered to be normal biological variation. Clinical chemistry of the female rats showed significantly reduced ALT and ALP in the experimental group. No gross lesions were observed during autopsy. No stillbirths were observed. Cesarean section rate and birth parameters (e.g., number of corpora lutea, number of implantations, number of live births) were not affected by the treatment.
The weight of male fetuses in the experimental group was significantly lower than that in the control group, but the difference in total fetal weight was not statistically significant and was less than 5%.
The overall incidence of fetal malformations (any observed malformation) in the experimental group (8% of the total number of fetuses) was significantly higher than that in the control group (4%). This increase was mainly due to skeletal variations, but the incidence of skeletal variations itself was not statistically different between the two groups. One fetus in the experimental group had a cleft palate (incomplete ossification). No specific teratogenic pattern associated with the test substance was found. [3]

[1]. Efficacy and safety of ketone ester infusion to prevent muscle weakness in a mouse model of sepsis-induced critical illness. Sci Rep. 2022 Jun 22;12(1):10591.

[2]. The Chemistry of the Ketogenic Diet: Updates and Opportunities in Organic Synthesis. Int J Mol Sci. 2021 May 15;22(10):5230.

[3]. Oral 28-day and developmental toxicity studies of (R)-3-hydroxybutyl (R)-3-hydroxybutyrate. Regul Toxicol Pharmacol. 2012 Jul;63(2):196-208.

(±)-1,3-Butanediol acetoacetate diester is a synthetic ketogenic compound. [2]
It is synthesized by transesterification of (±)-1,3-butanediol and tert-butylacetoacetate, a method published in 1995. [2]
Supplementing a normal diet with synthetic ketogenic compounds such as (±)-1,3-butanediol acetoacetate has been shown to induce ketosis and lower blood glucose levels without affecting triglyceride or cholesterol levels. [2]
It represents a potential alternative to a strict ketogenic diet for inducing nutritional ketosis. [2]

C8H16O4

176.21024

176.105

1208313-97-6

(S,S)-BD-AcAc 2;1251829-99-8

44631890

Typically exists as solid at room temperature

1.102

269 ºC

101 ºC

0.071

2

4

6

12

135

2

C[C@@H](O)CC(OCC[C@H](O)C)=O

AOWPVIWVMWUSBD-RNFRBKRXSA-N

InChI=1S/C8H16O4/c1-6(9)3-4-12-8(11)5-7(2)10/h6-7,9-10H,3-5H2,1-2H3/t6-,7-/m1/s1

[(3R)-3-hydroxybutyl] (3R)-3-hydroxybutanoate

BD-AcAc-2; BD-AcAc 2; BD AcAc 2

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~100 mg/mL (~567.50 mM)
DMSO : ~100 mg/mL (~567.50 mM)

Solubility in Formulation 1: ≥ 5.5 mg/mL (31.21 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 55.0 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.

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Solubility in Formulation 2: ≥ 5.5 mg/mL (31.21 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 55.0 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.

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Solubility in Formulation 3: ≥ 5.5 mg/mL (31.21 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 55.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 100 mg/mL (567.50 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)