Natural triterpenoid; Nrf2
Acetyl-11-keto-β-boswellic acid (AKBA) is known to induce and activate nrf2 .
AKBA induces the tyrosine phosphatase SHP-1. [1]

AKBA (Acetyl-11-keto-β-boswellic Acid) Significantly Reduces Infarct Volume and Improves Cellularity and Increases Neurological Score via Nrf2 and HO-1 Expression in 48-Hour Middle Cerebral Artery Occlusion (MCAO) Brain Tissue Reperfusion Recovery. In primary cultured neurons, AKBA increased the expression of Nrf2 and HO-1, thereby preventing OGD-induced oxidative damage. In addition, AKBA therapy improves the binding activity of Nrf2 to the antioxidant response element (ARE) [1]. AKBA (11-methyl-β-boswellic acid) strongly suppresses the development of human adrenal cancer cells, demonstrating cell cycle activation in the G1 phase and causing cellular inflammation [3]. AKBA (11-oneki-beta-boswellic acid) causes substantial lipolysis in 3T3-L1 adipocytes, as demonstrated by a decrease in cytoplasm neutral sodium and an increase in carotene in the intermediates. Increased lipolysis by AKBA is accompanied by upregulation of adipose lipase, adipocyte triglyceride lipase (ATGL) and adipose lipase (HSL), and decreased expression of the lipid droplet stability regulator perilipin. In addition, AKBA (Acetyl-11-keto-β-boswellic) Acid) treatment reduced the phenotypic markers of mature adipocytes aP2, adiponectin, and glut-4 in mature adipocytes [5].

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In Gie-3B11 human gingival epithelial cell layers, Acetyl-11-keto-β-boswellic acid (AKBA) (0.5 μM to 10 μM, 48-hour exposure to both cell surfaces) produced dual indications of enhanced barrier function: increased transepithelial electrical resistance (Rt) and decreased transepithelial mannitol flux (Jm). Specifically, 2 μM AKBA induced a maximal decrease in Jm (to 68.3% ± 5.4% of control) and an increase in Rt (to 120.4% ± 3.8% of control). 10 μM AKBA yielded the maximal increase in Rt (149.7% ± 6.0% of control) but induced leak to 14C-D-mannitol (Jm increased to 143.2% ± 12.3% of control). [1]
Western immunoblot analysis of total cell lysates from Gie-3B11 cell layers treated with 2 μM AKBA for 48 hours showed no significant change in claudin-1, claudin-2, or claudin-5 abundance, but induced a small (20%) but highly significant increase in claudin-4. Additionally, AKBA produced significant (30%) increases in claudin-3 and claudin-7 in Gie-3B11 cell layers (data not shown in figures but mentioned in text). [1]
Apical-only administration of AKBA (100 μM, 48 hours) did not decrease mannitol leak but produced over 50% increase in Rt. Apical-only AKBA required higher concentrations to produce similar effects on barrier function compared to exposure to both cell surfaces. For example, 50 μM AKBA apically increased Rt significantly, whereas lower concentrations (e.g., 2 μM) from both sides were effective. A concentration curve for apical-only AKBA (0.5 to 50 μM) showed that higher concentrations were needed for barrier enhancement. [1]

AKBA (acetyl-11-keto-beta-boswellic acid) strongly reduced intestinal adenomatous polyps without harm in mice. AKBA is more active than aspirin in preventing the growth of tiny intestinal and respiratory polyps. Histopathological investigation suggests that the effect of AKBA, i.e. reducing polyp size and degree of atypical repair, is more noticeable in polyps of larger size, especially those originating from intermittent polyps [1]. AKBA successfully inhibited the growth of HT-29 xenografts in mice without harm. AKBA is more active than aspirin [3]. AKBA demonstrates antitumor action both in vitro and in vivo. Through sidewall application in mice, AKBA significantly decreased SGC-7901 and MKN-45 xenografts without toxicity [4].

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Concentration-dependent effects of AKBA on Gie-3B11-barrier function. The cell layers were treated with AKBA on both cell surfaces for 48 h, beginning 4 days postseeding, before measurements were performed. Results shown represent the mean ± standard error for n = 7 cell layers (for the Rt) or n = 4 cell layers (for the mannitol flux). indicates P < 0.05, indicates P < 0.01, and indicates P < 0.001 (one-way ANOVA, Tukey).[1]
Cell layers were treated with 50 μM zinc, 400 μM quercetin, 15 μM retinoic acid, or 2 μM AKBA, as described. (A) Actual immunoblot bands observed for claudins-1, -2, -4, and -5 for the four control cell layers (lanes 1–4) and four micronutrient-treated cell layers (lanes 5–8). (B) Band densities quantitated by the optical densities.[1]

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Gie-3B11 cell culture and barrier function assessment: Gie-3B11 human gingival keratinocyte cells were seeded into sterile polycarbonate filter units (30 mm diameter, 0.4 μm pore size) at a density of 5×10^5 cells/4.2 cm² insert on day 0. On day 1, cells were refed with control medium containing penicillin (50 U/ml) and streptomycin (50 μg/ml). Nutraceutical treatments (including AKBA) were performed 3 days post-seeding, with exposure time of 48 hours. On the day of transepithelial permeability experiments, cell layers were refed with fresh control medium and incubated for 1.5-2 hours prior to electrophysiological readings. Transepithelial electrical resistance (Rt) was measured using 1 sec, 40 μA direct current pulses, and calculated using Ohm's law. After electrical measurements, basal-lateral medium was replaced with medium containing 0.1 mM, 0.1 μCi/ml 14C-D-mannitol and incubated at 37°C for 90 min. Mannitol flux rate (Jm) was calculated from scintillation counting of basal-lateral and apical samples, expressed as pmoles/min/cm². [1]
Immunoblot analysis of tight junction proteins: Gie-3B11 cells harvested from filter supports were washed in cold phosphate buffered saline, then lysed in lysis buffer with protease and phosphatase inhibitors. Whole cell lysates were prepared by sonication and ultracentrifugation. Samples were analyzed by PAGE using 4%-20% gradient Tris-glycine gel at 125V for 1 hour 40 minutes. Proteins were transferred to PVDF membrane at 30V for 2 hours. Membranes were blocked with 5% milk/PBS-T overnight at 4°C, then incubated with primary antibody (anti-claudin-1, -2, -4, -5) at 0.5 μg/ml in 5% milk/PBS-T for 2 hours at room temperature, followed by secondary antibody (HRP-labeled) for 1 hour. Chemiluminescence detection was performed, and band densities were quantified. [1]
Confocal immunofluorescence: Gie-3B11 cells cultured on glass coverslips were fixed in ice-cold acetone for 5 minutes, blocked with 5% normal goat serum in PBS/0.1% Tween 20 for 30 minutes, then treated with primary antibody (mouse anti-human claudin-2; 5 μg/ml) for 45 minutes at room temperature, followed by Cy3-conjugated secondary antibody (1:500) for 45 minutes. Nuclei were stained with DAPI and cells were mounted and viewed under confocal microscopy. For AKBA, this method was used to assess changes in claudin localization (though no AKBA-specific immunofluorescence images were shown in the paper, the method was described for quercetin and applies to the same cell line). [1]

[1]. J Agric Food Chem. 2017 Dec 7. doi: 10.1021/acs.jafc.7b04203.

3-Acetyl-11-keto-β-boswellic acid is a triterpenoid compound. It has been reported to exist in Boswellia sacra, Boswellia serrata, and Boswellia papyrifera, with relevant data available. See also: Indian Boswellia (partial).

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Acetyl-11-keto-β-boswellic acid (AKBA) has been shown earlier to support barrier function in human gastrointestinal epithelial cell layers by decreasing the deleterious effects of hydrogen peroxide, TNF-α, and interferon-γ. This protective action was accompanied by a decrease in the level of NF-κB phosphorylation produced by proinflammatory stimuli. AKBA has also been shown to inhibit leukotriene biosynthesis, as well as the synthesis and secretion of TNF-α, IL-1, IL-2, IL-6, and interferon-γ. [1]
The ability of AKBA to induce compositional changes in tight junction complexes (e.g., increasing claudin-4, -3, -7) may have implications for preventing pathogen binding and translocation across epithelial barriers, as certain pathogens bind to specific claudins. AKBA may be prophylactically effective in reducing the frequency and/or severity of certain infections by modifying the claudin "neighborhood" in tight junctions. [1]
AKBA was effective when administered apically only (as an oral spray or lozenge would be in clinical use), though higher concentrations were required compared to basal-lateral exposure. This is relevant for topical oral prophylaxis to improve oral epithelial barrier function. [1]

C32H48O5

512.73

512.35

C, 74.96; H, 9.44; O, 15.60

67416-61-9

11168203

White to off-white solid powder

1.1±0.1 g/cm3

600.3±55.0 °C at 760 mmHg

184.4±25.0 °C

0.0±3.7 mmHg at 25°C

1.549

8

1

5

3

37

1060

11

C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC(=O)[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@H]([C@]5(C)C(=O)O)OC(=O)C)C)C)[C@@H]2[C@H]1C)C)C

HMMGKOVEOFBCAU-BCDBGHSCSA-N

InChI=1S/C32H48O5/c1-18-9-12-28(4)15-16-30(6)21(25(28)19(18)2)17-22(34)26-29(5)13-11-24(37-20(3)33)32(8,27(35)36)23(29)10-14-31(26,30)7/h17-19,23-26H,9-16H2,1-8H3,(H,35,36)/t18-,19+,23-,24-,25+,26-,28-,29+,30-,31-,32-/m1/s1

(3R,4R,4aR,6aR,6bS,8aR,11R,12S,12aR,14aR,14bS)-3-acetyloxy-4,6a,6b,8a,11,12,14b-heptamethyl-14-oxo-1,2,3,4a,5,6,7,8,9,10,11,12,12a,14a-tetradecahydropicene-4-carboxylic acid

3-O-acetyl-11-keto-β-Boswellic acid; 3-acetyl-11-keto-β-Boswellic acid; AKBA; 67416-61-9; 3-acetyl-11-keto-beta-boswellic acid; 3-O-Acetyl-11-keto-beta-Boswellic Acid; AKBA cpd; acetyl-11-ketoboswellic acid; Acetyl-11-keto-beta-boswellic acid; UNII-BS16QT99Q1; Acetylketo-β-boswellic acid

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)

DMSO : ≥ 5.2 mg/mL (~10.14 mM)

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.

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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).

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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)

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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).

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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

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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.

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