Liquid chromatography-mass spectrometry (LC-MS) metabolic profiling of media from primary mouse myocytes with forced expression of PGC-1α revealed a 2.7-fold increase in the secretion of β-aminoisobutyric acid (BAIBA) compared to control GFP-expressing cells. [1]
Treatment of primary mouse adipocytes differentiated from the stromal vascular fraction of inguinal white adipose tissue (WAT) with 5 µM BAIBA for 6 days significantly increased the mRNA expression of brown adipocyte-specific genes UCP-1 (5.3-fold) and CIDEA (2.25-fold) in a dose-dependent manner, without altering the expression of the white adipocyte gene ADIPOQ. [1]
Treatment of human induced pluripotent stem cell (iPSC)-derived white adipocytes with 1 µM or 10 µM BAIBA during differentiation induced a dose-dependent increase in the expression of brown adipocyte-specific genes (UCP-1, CIDEA, PRDM16, ELOVL3). [1]
In the same human iPSC-derived white adipocytes, treatment with BAIBA (concentration not specified for this assay) significantly increased both basal and insulin-stimulated glucose uptake. It also increased the basal oxygen consumption rate (OCR), the oligomycin-insensitive OCR (indicative of uncoupling), and the maximal respiratory capacity induced by carbonyl cyanide m-chlorophenylhydrazone (CCCP). [1]
Treatment of mouse primary adipocytes with 5 µM BAIBA increased the mRNA expression of PPARα by 2.4-fold. This effect, along with the BAIBA-induced upregulation of UCP-1 and CIDEA, was abolished by co-treatment with the selective PPARα antagonist GW6471 or by using adipocytes derived from PPARα null mice. [1]
Treatment of H4IIE hepatocytes with 5 µM BAIBA for 6 days significantly increased the mRNA expression of PPARα (5.4-fold) and key fatty acid β-oxidation genes (CPT1, ACADvI, ACADm, ACOX1). This induction was abolished by co-treatment with the PPARα antagonist GW6471. [1]
Treatment of H4IIE hepatocytes with BAIBA at concentrations ranging from 0.1 to 10 µM for 6 days dose-dependently increased the maximal oxygen consumption rate (OCR) induced by the uncoupler FCCP. [1]

Following treatment with 3-amino-2-methylpropionic acid (BAIBA), the mice's body weight decreased somewhat. MRI-based body composition study revealed that BAIBA therapy dramatically decreased mice's body fat. Metabolic cage study revealed that oxygen consumption (VO2) and whole-body energy expenditure were raised in BAIBA-treated mice without any significant variations in activity or food intake, which is consistent with the effects on thermogenic and beta-oxidation gene expression and body weight. Additionally, mice were given an intraperitoneal glucose tolerance test (IPGTT) challenge. The area under the IPGTT curve indicates that BAIBA can dramatically increase mice's glucose tolerance [1].

---

Oral administration of BAIBA (100 mg/kg/day or 170 mg/kg/day) in drinking water to C57BL6/J mice for 14 days dose-dependently increased plasma BAIBA concentrations (to 2 µM and 8.9 µM, respectively) and significantly upregulated the expression of brown adipocyte-specific genes (UCP-1, CIDEA, PGC-1α, Cytochrome C) in inguinal white adipose tissue (WAT). [1]
Muscle-specific PGC-1α transgenic (MCK-PGC1α) mice showed an 11-fold increase in plasma BAIBA concentration (6.5 µM) compared to controls. Conversely, PGC-1α knockout mice had reduced plasma BAIBA levels. [1]
Wild-type mice subjected to 3 weeks of voluntary wheel running (exercise) showed a significant increase in plasma BAIBA concentration (19% increase to 2.6 µM) compared to sedentary controls. [1]
Oral administration of BAIBA (100 mg/kg/day) in drinking water to C57BL6/J mice for 14 weeks (while on a high-fat diet) resulted in slightly decreased body weight gain, significantly reduced body fat percentage, increased whole-body oxygen consumption (VO2) and energy expenditure (without changes in activity or food intake), and improved glucose tolerance as measured by intraperitoneal glucose tolerance test (IPGTT). [1]
The induction of brown adipocyte-specific gene expression (UCP-1, CIDEA, PGC-1α, Cytochrome C) in inguinal WAT by BAIBA treatment (100 mg/kg/day for 14 days) was completely abolished in PPARα null mice. [1]
Oral administration of BAIBA (100 mg/kg/day) in drinking water to C57BL6/J mice for 14 days significantly increased the hepatic mRNA expression of PPARα and key β-oxidation genes (CPT1, ACADvI, ACADm, ACOX1). This induction was absent in PPARα null mice. [1]

Myocyte Culture and Metabolite Profiling: Primary mouse myoblasts were differentiated into myotubes. On day 2 of differentiation, cells were transduced with an adenovirus expressing either PGC-1α or GFP. After 24 hours, the media was replaced with serum-free media. Following another 24-hour incubation, the conditioned media was collected, centrifuged, and the supernatant was analyzed using an LC-MS-based metabolic profiling platform to quantify small molecule metabolites, including BAIBA. [1]
Primary Mouse Adipocyte Differentiation and Treatment: The stromal vascular fraction was isolated from mouse inguinal white adipose tissue. Cells were cultured and induced to differentiate into adipocytes using a differentiation cocktail containing insulin, dexamethasone, IBMX, indomethacin, T3, and rosiglitazone. During the 6-day differentiation process, cells were treated with vehicle or various concentrations of BAIBA (e.g., 0.3, 1, 3, 5 µM). For inhibition studies, the PPARα antagonist GW6471 was added. RNA was extracted at the end of differentiation for gene expression analysis by quantitative PCR (qPCR). [1]
Human iPSC Differentiation into Adipocytes: Human induced pluripotent stem cells (iPSCs) were differentiated into mesenchymal progenitor cells, which were then programmed to become white adipocytes via lentiviral expression of PPARG2. During the 21-day adipogenic differentiation process, the medium was supplemented with or without BAIBA (1 µM or 10 µM). Gene expression was analyzed by qPCR. [1]
Glucose Uptake Assay in Human iPSC-Derived Adipocytes: Differentiated adipocytes were serum-starved and then incubated in a Krebs-Ringer HEPES buffer. Glucose uptake was measured by incubating cells with radiolabeled 2-deoxy-D-glucose for 5 minutes. Nonspecific uptake was determined in the presence of cytochalasin B and subtracted. [1]
Cellular Oxygen Consumption Rate (OCR) Measurement: Differentiated adipocytes or hepatocytes were plated in specialized microplates. The OCR was measured using an extracellular flux analyzer. Mitochondrial function was assessed by sequential injection of oligomycin (ATP synthase inhibitor), CCCP or FCCP (mitochondrial uncouplers), and antimycin A (complex III inhibitor). [1]
Hepatocyte Culture and Treatment: H4IIE hepatocytes were seeded in collagen-coated plates and cultured in standard medium supplemented with or without 5 µM BAIBA and/or 1 µM GW6471 (PPARα antagonist) for 6 days, with medium changes every two days. RNA was then extracted for qPCR analysis of β-oxidation genes. [1]
Hepatocyte Respirometry (OxoPlate): H4IIE hepatocytes were plated in 96-well plates containing an oxygen-sensitive fluorescent sensor at the bottom. Cells were treated with a range of BAIBA concentrations (0-10 µM) with or without GW6471 for 6 days, with regular medium changes. After 6 days, the medium was replaced with serum-free medium containing the same compounds and overlaid with mineral oil. The uncoupler FCCP was added to induce maximal respiration. Fluorescence was measured at multiple time points to calculate oxygen consumption rates. [1]

BAIBA Treatment in Wild-type and PPARα Null Mice: Six-week-old C57BL6/J (wild-type) or PPARα null mice were weight-matched and assigned to control or treatment groups. BAIBA was dissolved in the drinking water at a dose of 100 mg/kg/day or 170 mg/kg/day. Mice had ad libitum access to this medicated water (and either standard chow or a high-fat diet, as specified) for 14 days or 14 weeks. Control mice received normal drinking water. [1]
Tissue Collection: At the end of the treatment period, mice were fasted, euthanized, and plasma was collected via cardiac puncture. Tissues of interest (e.g., inguinal white adipose tissue, liver) were rapidly dissected, snap-frozen in liquid nitrogen, and stored for subsequent RNA extraction and gene expression analysis. [1]
Indirect Calorimetry: Mice treated with BAIBA (100 mg/kg/day) or water control for 14 weeks were placed in metabolic cages (PhenoMaster system). After a 7-day acclimation period, parameters including oxygen consumption (VO2), carbon dioxide production (VCO2), food intake, body mass, and locomotor activity were monitored twice hourly for several days under a controlled light/dark cycle with ad libitum access to food and (medicated) water. [1]
Intraperitoneal Glucose Tolerance Test (IPGTT): After a 6-hour fast, mice were injected intraperitoneally with a glucose solution (1.5 mg glucose per gram of body weight). Blood glucose levels were measured from the tail vein immediately before injection (0 min) and at 30, 60, and 120 minutes post-injection using a glucometer. The area under the curve (AUC) was calculated to assess glucose tolerance. [1]
Exercise Training in Mice: Twelve-week-old wild-type mice were given access to free-running wheels for 3 weeks to perform voluntary endurance exercise. Sedentary control mice were housed without running wheels. After the training period, plasma and muscle tissues were collected for metabolite analysis. [1]

[1]. β-Aminoisobutyric acid induces browning of white fat and hepatic β-oxidation and is inversely correlated with cardiometabolic risk factors. Cell Metab. 2014 Jan 7;19(1):96-108.

[2]. BAIBA attenuates insulin resistance and inflammation induced by palmitate or a high fat diet via an AMPK-PPARδ-dependent pathway in mice. Diabetologia. 2015 Sep;58(9):2096-105.

[3]. β-Aminoisobutyric acid (L-BAIBA) is a novel regulator of mitochondrial biogenesis and respiratory function in human podocytes. Sci Rep. 2023 Jan 14;13(1):766.

[4]. Signaling metabolite β-aminoisobutyric acid as a metabolic regulator, biomarker, and potential exercise pill. Front Endocrinol (Lausanne). 2023 May 29;14:1192458.

3-Aminoisobutyric acid (3-aminoisobutyric acid) is a β-amino acid, a product of isobutyric acid with one methyl hydrogen atom replaced by an amino group. It is a metabolite functionally related to isobutyric acid. It is the conjugate acid of 3-aminoisobutyrate ester and also a zwitterion tautomer of 3-aminoisobutyrate ester. 3-Az-2-methylpropionate has been reported in Drosophila, Salmonella enteritidis, and Caenorhabditis elegans, with relevant data available. 3-Aminoisobutyric acid is a derivative of the amino acid alanine, a product of thymine metabolism, and has the potential to induce downstream metabolic activities. During exercise, 3-aminoisobutyric acid is secreted into the bloodstream by muscle cells. Once this molecule interacts with adipocytes, it may induce signaling pathways regulating lipid, insulin, and cholesterol metabolism. β-Aminoisobutyric acid (BAIBA) has been identified as a novel small molecule myocytokine secreted by skeletal muscle cells in response to exercise-induced increases in the expression of the transcriptional coactivator PGC-1α. [1]
In a large community cohort study (Framingham Heart Study, n=2067), plasma BAIBA levels were significantly negatively correlated with cardiovascular metabolic risk factors (fasting blood glucose, insulin, HOMA-IR, triglycerides, total cholesterol). [1]
In a controlled exercise intervention study (HERITAGE, n=80), 20 weeks of endurance training significantly increased plasma BAIBA concentration by 17%. [1] Human genetic association studies have found that single nucleotide polymorphisms (SNPs) in genes involved in valine catabolism and BAIBA biosynthesis pathways (e.g., AGXT2, ACADS, HADHA) are closely associated with plasma BAIBA levels. Studies have also found that several genes in these pathways are upregulated by PGC-1α transcription in muscle cells. [1] This study proposes that BAIBA is a PGC-1α-dependent exercise-induced signal that promotes the interaction between muscle and adipose tissue/liver, thereby promoting beneficial metabolic phenotypes (browning of white adipose tissue, enhanced hepatic β-oxidation) and improving glucose homeostasis, and may mediate some protective effects of exercise against metabolic diseases. [1]

C4H9NO2

103.1198

103.063

144-90-1

(S)-b-aminoisobutyric acid;4249-19-8;3-Amino-2-methylpropanoic acid-d3;1219803-65-2

64956

White to off-white solid powder

1.1±0.1 g/cm3

223.6±23.0 °C at 760 mmHg

246.72°C (estimate)

89.0±22.6 °C

0.0±0.9 mmHg at 25°C

1.462

-0.51

2

3

2

7

72.1

0

QCHPKSFMDHPSNR-UHFFFAOYSA-N

InChI=1S/C4H9NO2/c1-3(2-5)4(6)7/h3H,2,5H2,1H3,(H,6,7)

3-amino-2-methylpropanoic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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 (~969.74 mM)
DMSO : ~1 mg/mL (~9.70 mM)

Solubility in Formulation 1: 100 mg/mL (969.74 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

---

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