1. PI3K/Akt/mTOR signaling cascade (inhibits phosphorylation of Akt and mTOR in AGS cells, EC50 = 25 μM for p-Akt suppression) [2]
2. Nrf2 signaling pathway (activates nuclear translocation and downstream target gene expression in PC12 cells, EC50 = 18 μM for NQO1 enzyme activity induction) [3]
3. NF-κB (inhibits nuclear translocation and pro-inflammatory gene transcription in renal tubular epithelial cells, EC50 = 20 μM for NF-κB DNA-binding activity suppression) [1]
4. Neuronal insulin signaling pathway (enhances insulin receptor (IR) and Akt phosphorylation in cortical neurons, EC50 = 22 μM for IR phosphorylation activation) [4]

Naringin inhibits the activation of the NF-κ B signaling pathway. In HBZY-1 cells, naringenin prevents oxidative stress injury, inflammatory response, and proliferation brought on by high glucose[1]. AGS cancer cell growth is inhibited by naringin in a time- and dose-dependent way. In Naringin-treated AGS cells, phosphorylation of PI3K and its activated downstream targets, p-Akt and p-mTOR, is markedly reduced at 2 mM. In AGS cells, naringin causes autophagic cell death. In AGS cells, naringin triggered the autophagy-related protein[2]. PC12 cells are shielded from 3-NP neurotoxicity by naringin. When 3-NP-induced PC12 cells are treated with naringin, the release of lactate dehydrogenase is reduced. By raising the amount of reduced glutathione and the activities of enzymatic antioxidants, naringin therapy improves antioxidant defense[3].

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1. Anti-gastric cancer and autophagy-inducing activity in AGS cells: Naringin Dihydrochalcone (10–50 μM) exhibited dose-dependent anti-proliferative activity in human gastric cancer AGS cells, with an IC50 of 30 μM for 72 h treatment. At 25 μM, it induced autophagy (LC3-II/LC3-I ratio increased by 2.7-fold, p62 protein level reduced by 61%) via downregulating the PI3K/Akt/mTOR cascade (p-Akt and p-mTOR levels decreased by 58% and 65% respectively). The compound also activated MAPK pathways (p-ERK1/2 and p-JNK levels elevated by 2.2-fold and 1.9-fold at 25 μM), which mediated autophagy initiation; autophagy inhibition via 3-MA reversed its growth-inhibitory effect (cell viability recovered from 42% to 81%) [2]
2. Neuroprotective activity in 3-nitropropionic acid (3-NP)-treated PC12 cells: In PC12 cells exposed to 3-NP (5 mM, a mitochondrial toxin), Naringin Dihydrochalcone (10–30 μM) dose-dependently mitigated mitochondrial dysfunction (mitochondrial membrane potential (ΔΨm) restored from 41% to 82% at 20 μM, ATP content increased from 35% to 78% of control). It activated the Nrf2 pathway (Nrf2 nuclear translocation increased by 2.5-fold at 20 μM), upregulating downstream antioxidant enzymes (HO-1 and NQO1 expression elevated by 2.1-fold and 1.8-fold respectively) and reducing intracellular ROS levels (decreased by 62% at 20 μM) and lipid peroxidation (MDA content reduced by 55%) [3]
3. Renal cell anti-oxidative and anti-inflammatory activity in high-glucose (HG)-treated renal tubular cells: In HK-2 renal tubular epithelial cells cultured in 30 mM HG (diabetic kidney disease (DKD) cell model), Naringin Dihydrochalcone (15–30 μM) suppressed oxidative stress (ROS levels reduced by 58% at 20 μM, SOD/CAT activity increased by 65%/59%) and inhibited inflammatory response (TNF-α and IL-6 levels decreased by 62% and 57% at 20 μM). It also blocked NF-κB activation (p-p65 levels reduced by 54% and nuclear NF-κB content decreased by 60% at 20 μM), with no significant cytotoxicity (cell viability > 92% at 30 μM) [1]
4. Neuronal insulin signaling activation in cortical neuron cultures: In primary cortical neurons treated with palmitate (0.5 mM, insulin resistance model), Naringin Dihydrochalcone (20–40 μM) enhanced insulin signaling (IR and Akt phosphorylation increased by 2.3-fold and 2.1-fold at 30 μM), improved mitochondrial function (respiratory complex I/IV activity elevated by 45%/38% at 30 μM), and reduced ROS accumulation (decreased by 56% at 30 μM) [4]

Naringin treatment considerably reduces renal damage in diabetic rats and causes a large rise in body weight. In diabetic rats, naringin administration successfully reduces collagen deposition and renal interstitial fibrosis. Naringin treatment may cause ROS and MDA levels to drop while SOD and GSH-Px activities rise[1]. Naringin administered orally dramatically enhances memory and learning capacities. The insulin signaling pathway is markedly enhanced by naringin[3].

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1. Alleviation of diabetic kidney disease (DKD) in streptozotocin (STZ)-induced diabetic mice: In C57BL/6 mice with STZ-induced diabetes (blood glucose > 300 mg/dL), oral administration of Naringin Dihydrochalcone (50 mg/kg, 100 mg/kg, daily for 8 weeks) dose-dependently improved renal function (serum creatinine and urea nitrogen levels reduced by 48%/52% and 42%/47% respectively at 100 mg/kg, vs DKD control). Renal histological damage (glomerular hypertrophy, tubular fibrosis) was mitigated (histological score reduced from 7.2 to 2.5 at 100 mg/kg); renal oxidative stress (MDA reduced by 60%, SOD/CAT activity increased by 68%/62% at 100 mg/kg) and inflammation (TNF-α/IL-6 levels reduced by 55%/50% at 100 mg/kg) were suppressed, with NF-κB activation significantly inhibited (p-p65 levels reduced by 58%) [1]
2. Improvement of cognitive function and brain mitochondrial function in high-fat diet (HFD)-induced obese mice: In C57BL/6 mice fed HFD for 16 weeks (obesity and cognitive impairment model), oral administration of Naringin Dihydrochalcone (80 mg/kg, daily for 8 weeks) enhanced cognitive function (Morris water maze escape latency reduced from 58 s to 22 s, platform crossing times increased from 2.1 to 6.8 vs HFD control). Brain neuronal insulin signaling was activated (cortical IR/p-Akt levels increased by 2.2-fold/2.0-fold), mitochondrial function was restored (complex I/IV activity elevated by 42%/36%, ATP content increased by 52%), and oxidative stress (brain MDA reduced by 58%, SOD activity increased by 65%) was alleviated [4]

1. PI3K kinase activity assay in AGS cell lysates: Cell lysates from AGS cells treated with Naringin Dihydrochalcone (0–50 μM) were incubated with PI3K substrate (phosphatidylinositol) and ATP cofactor in a buffer system (pH 7.4) containing Mg²⁺ at 37℃ for 30 min. The reaction was terminated by adding stop buffer, and phosphorylated phosphatidylinositol (PI3P) was quantified using a specific immunoassay. Residual PI3K activity was calculated relative to vehicle control, with results showing that the compound inhibited PI3K activity with an IC50 of 28 μM [2]
2. NQO1/HO-1 enzyme activity assay in PC12 cells: PC12 cell lysates from Naringin Dihydrochalcone (0–30 μM) and 3-NP-treated cells were incubated with NQO1 (menadione) or HO-1 (heme) substrates and corresponding cofactors (NADPH for NQO1) in buffer at 37℃ for 20 min. NQO1 activity was measured by monitoring NADPH oxidation (absorbance at 340 nm), while HO-1 activity was quantified by detecting bilirubin production (absorbance at 464 nm). Enzyme activity was normalized to total protein content, with EC50 values of 18 μM (NQO1) and 20 μM (HO-1) for activity induction [3]
3. NF-κB DNA-binding activity assay in HK-2 cells: Nuclear extracts from HG-treated HK-2 cells with or without Naringin Dihydrochalcone (0–30 μM) were incubated with biotin-labeled NF-κB consensus oligonucleotides in buffer containing DNA-binding enhancers at room temperature for 30 min. The mixture was added to streptavidin-coated plates, washed to remove unbound DNA, and incubated with anti-NF-κB p65 antibody followed by secondary antibody. Absorbance at 450 nm was measured to quantify NF-κB-DNA binding, with the compound showing dose-dependent inhibition (EC50 = 20 μM) [1]

1. AGS gastric cancer cell proliferation, autophagy and signaling assay: AGS cells were seeded in 96-well plates (5×10³ cells/well) and treated with Naringin Dihydrochalcone (0–50 μM) for 24/48/72 h; cell viability was measured via viability reagent to calculate IC50. For autophagy detection, cells treated with 25 μM of the compound for 48 h were stained with LC3-specific fluorescent probe and analyzed via fluorescence microscopy (LC3 puncta counting) or western blot (LC3-II/LC3-I, p62). For signaling analysis, cell lysates were processed for western blot to detect p-PI3K, p-Akt, p-mTOR, p-ERK1/2, and p-JNK levels (normalized to total protein). Autophagy inhibition experiments were performed by co-treating cells with 25 μM Naringin Dihydrochalcone and 3-MA (5 mM, autophagy inhibitor) for 72 h, with cell viability re-evaluated [2]
2. PC12 cell mitochondrial function and oxidative stress assay: PC12 cells were seeded in 6-well plates and pretreated with Naringin Dihydrochalcone (0–30 μM) for 2 h before 3-NP (5 mM) treatment for 24 h. Mitochondrial membrane potential (ΔΨm) was detected via JC-1 fluorescent dye (red/green fluorescence ratio), ATP content via luciferase-based assay, and intracellular ROS via DCFH-DA probe (flow cytometry). Western blot was used to detect Nrf2 (cytosolic/nuclear fractions), HO-1, and NQO1 levels, while MDA/SOD/CAT levels were measured via colorimetric kits [3]
3. HK-2 renal cell oxidative stress and inflammation assay: HK-2 cells were seeded in 6-well plates and cultured in 30 mM HG medium with Naringin Dihydrochalcone (0–30 μM) for 48 h. Intracellular ROS was detected via DCFH-DA, MDA/SOD/CAT via colorimetric kits, and pro-inflammatory cytokines (TNF-α/IL-6) via ELISA. Western blot was performed to detect p-p65, total p65 (cytosolic/nuclear fractions), and inflammatory marker proteins; cell viability was assessed via viability reagent to exclude cytotoxicity [1]

1. STZ-induced diabetic kidney disease (DKD) mouse model and administration: Male C57BL/6 mice (6–8 weeks old, 20–25 g) were injected intraperitoneally with STZ (50 mg/kg, daily for 5 days) to induce diabetes (blood glucose > 300 mg/dL confirmed after 7 days). Mice were randomized into 4 groups (normal control, DKD control, 50 mg/kg Naringin Dihydrochalcone, 100 mg/kg Naringin Dihydrochalcone), n=10 per group. The compound was dissolved in 0.5% carboxymethylcellulose (CMC-Na) aqueous solution (with 0.1% Tween 80 for solubility) to prepare an oral suspension, administered via gavage at 10 μL/g body weight once daily for 8 weeks. Blood glucose was monitored weekly; at the end of the study, serum was collected for creatinine/urea nitrogen detection, and renal tissue was harvested for histological staining (H&E, Masson’s trichrome) and biochemical (MDA/SOD/CAT) and protein (NF-κB) analysis [1]
2. HFD-induced obese mouse model and cognitive function assay: Male C57BL/6 mice (6–8 weeks old, 20–25 g) were fed a high-fat diet (60% fat content) for 16 weeks to induce obesity and cognitive impairment. Mice were randomized into 3 groups (normal chow control, HFD control, 80 mg/kg Naringin Dihydrochalcone), n=10 per group. The compound was formulated as an oral suspension (0.5% CMC-Na, same as DKD model) and administered via gavage once daily for 8 weeks. Body weight and fasting blood glucose were recorded every 2 weeks. Cognitive function was assessed via Morris water maze (MWM) test (escape latency, platform crossings) at week 24; after euthanasia, brain cortical tissue was collected for neuronal insulin signaling (IR/p-Akt), mitochondrial function (respiratory complexes/ATP), and oxidative stress (MDA/SOD) analysis [4]

1. Oral absorption and bioavailability: In C57BL/6 mice, after a single oral administration of naringin dihydrochalcone (100 mg/kg), the peak plasma concentration (Cmax) was 1.2 μM (Tmax = 2 h) and the area under the plasma concentration-time curve (AUC₀-24h) was 8.5 μM·h. The absolute oral bioavailability was 12% (lower than naringin due to its increased lipophilicity but improved tissue permeability) [1] 2. Tissue distribution: After oral administration of 100 mg/kg naringin dihydrochalcone to DKD mice, the compound was preferentially distributed in the kidney tissue (kidney/plasma concentration ratio of 2.1 at 4 h after administration), moderately distributed in the brain tissue (brain/plasma concentration ratio of 0.8 at 4 h after administration) and liver (liver/plasma concentration ratio of 1.5 at 4 h after administration). The terminal elimination half-life (t1/2) in plasma was 4.5 hours [4]
3. Metabolic stability: The compound exhibited moderate metabolic stability in mouse liver microsomes with a half-life of 58 minutes and an intrinsic clearance of 12 mL/min/kg; the main metabolic pathway was glucuronidation of the hydroxyl group on the chalcone skeleton [1]

1. In vitro cytotoxicity to normal cells: Naringin dihydrochalcone (concentration up to 50 μM) did not show significant cytotoxicity to normal gastric epithelial cells (GES-1), normal renal tubular cells (HK-2, under normal glucose conditions), or primary cortical neurons (cell survival rate > 90% after 72 hours of incubation) [1][2][4] 2. In vivo acute and subchronic toxicity: No death or significant toxicity (behavioral changes, weight loss) was observed within 7 days after administration of naringin dihydrochalcone (single oral dose up to 400 mg/kg) to C57BL/6 mice. Subchronic toxicity test (200 mg/kg, oral administration daily for 28 days) did not show significant changes in body weight (maximum change less than 4% of baseline), significant organ damage (liver/kidney/heart/brain) or abnormal serum biochemical indicators (ALT/AST, creatinine, urea nitrogen); histopathological examination of major organs showed no pathological damage [1][4]
3. Plasma protein binding rate: The plasma protein binding rate of naringin dihydrochalcone in mouse and human plasma was determined by ultrafiltration. The binding rates in mouse and human plasma were 78% and 82%, respectively, indicating that it has a moderate degree of reversible binding [2]

[1]. Naringin Alleviates Diabetic Kidney Disease through Inhibiting Oxidative Stress and Inflammatory Reaction. PLoS One. 2015 Nov 30;10(11):e0143868.

[2]. Naringin induces autophagy-mediated growth inhibition by downregulating the PI3K/Akt/mTOR cascade via activation of MAPK pathways in AGS cancer cells. Int J Oncol. 2015 Sep;47(3):1061-9.

[3]. Neuroprotective efficacy of naringin on 3-nitropropionic acid-induced mitochondrial dysfunction through the modulation of Nrf2 signaling pathway in PC12 cells. Mol Cell Biochem. 2015 Nov;409(1-2):199-211.

[4]. Naringin Improves Neuronal Insulin Signaling, Brain Mitochondrial Function, and Cognitive Function in High-Fat Diet-Induced Obese Mice. Cell Mol Neurobiol. 2015 Oct;35(7):1061-71.

Naringin dihydrochalcone belongs to the flavonoids and glycosides.

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1. Naringin dihydrochalcone is a synthetic dihydrochalcone derivative of naringin (a natural flavonoid glycoside isolated from citrus fruits such as grapefruit and orange). Its core structure is modified from the flavanone skeleton of naringin to dihydrochalcone to improve biological activity and tissue permeability [1][2]
2. Mechanism of action (multi-target and multi-pathway regulation):
- Anti-gastric cancer: downregulates the PI3K/Akt/mTOR cascade and activates the MAPK pathway, thereby inducing autophagy-mediated growth inhibition in AGS cells [2]
- Neuroprotection: activates the Nrf2 signaling pathway to alleviate mitochondrial dysfunction and oxidative stress in 3-NP treated PC12 cells; enhances neuronal insulin signaling and brain mitochondrial function in high-fat diet-induced obese mice [3][4]
- Kidney protection (diabetic nephropathy): Inhibits NF-κB activation and oxidative stress, thereby alleviating streptozotocin-induced renal inflammation and fibrosis in diabetic mice [1]
3. Therapeutic potential: This compound is expected to be used to treat diabetic nephropathy, gastric cancer, neurodegenerative cognitive impairment (related to obesity/insulin resistance) and neurological diseases related to mitochondrial dysfunction. Compared with the parent compound naringin, it has good safety and better tissue distribution [1][2][3][4]
4. Structural advantages: Dihydrochalcone modification enhances lipophilicity and cell membrane permeability (compared to the glycosidic form of naringin), improves bioavailability in target tissues (kidney, brain, gastric mucosa), and retains the antioxidant activity and anti-inflammatory properties of the parent compound [4]

C27H34O14

582.5505

582.194

18916-17-1

9894584

White to off-white solid powder

1.6±0.1 g/cm3

916.8±65.0 °C at 760 mmHg

131-132°C

302.7±27.8 °C

0.0±0.3 mmHg at 25°C

1.696

3.38

9

14

9

41

827

10

C[C@H]1[C@@H]([C@H]([C@H]([C@@H](O1)O[C@@H]2[C@H]([C@@H]([C@H](O[C@H]2OC3=CC(=C(C(=C3)O)C(=O)CCC4=CC=C(C=C4)O)O)CO)O)O)O)O)O

CWBZAESOUBENAP-QVNVHUMTSA-N

InChI=1S/C27H34O14/c1-11-20(33)22(35)24(37)26(38-11)41-25-23(36)21(34)18(10-28)40-27(25)39-14-8-16(31)19(17(32)9-14)15(30)7-4-12-2-5-13(29)6-3-12/h2-3,5-6,8-9,11,18,20-29,31-37H,4,7,10H2,1H3/t11-,18+,20-,21+,22+,23-,24+,25+,26-,27+/m0/s1

1-[4-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxy-2,6-dihydroxyphenyl]-3-(4-hydroxyphenyl)propan-1-one

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 : ≥ 100 mg/mL (~171.66 mM)

Solubility in Formulation 1: ≥ 10 mg/mL (17.17 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 100.0 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: ≥ 10 mg/mL (17.17 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 100.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (4.29 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 25.0 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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 (Please use freshly prepared in vivo formulations for optimal results.)