- Estrogen receptor β (ERβ) (EC₅₀: 0.1–0.5 μM in reporter gene assays)
- Sirtuin 1 (SIRT1) (activation at 10–20 μM in vitro)

甘草素用 ERβ 而不是 ERα 转染 U2OS 细胞，这会导致 ERE tk-Luc 的剂量反应性激活。除了剂量激活 ERβ（而非 ERα）之外，甘草素还引起 CECR6、NKG2E 和 NKD 的暂时兴奋。甘草素的 ERβ 选择性是由于共激活因子休克共激活因子样 2 被选择性地招募到目标基因所致。 Liquiritigenin 以相当的亲和力与 ERα 和 ERβ 结合，从而导致 SRC-2 选择性募集到 ERβ 细胞中的靶基因 [Liquiritigenin MC3T3-E1 细胞抑制 TNF-α、细胞内活性氧、蛋白质加合物、MG-诱导成骨细胞 MC3T3-E1 死亡、线粒体超氧化物和心磷脂过氧化 [1, 2]。

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- ERβ介导的转录激活：在转染了ERβ表达质粒和ERβ响应性荧光素酶报告质粒的HEK293细胞中，Liquiritigenin 剂量依赖性激活报告基因。该激活作用的EC₅₀为0.3 μM，在1 μM时达到最大激活（相对于溶媒组激活2.8倍）。值得注意的是，该化合物即使在浓度高达10 μM时也不激活ERα，显示出对ERβ的高选择性。此外，与ERβ特异性拮抗剂PHTPP（1 μM）共同处理可完全消除Liquiritigenin（1 μM）的激活效应，证实其活性依赖于ERβ[1]
- 保护MC3T3-E1细胞对抗甲基乙二醛（MGO）诱导的细胞毒性：通过MTT实验检测，Liquiritigenin（5、10、20 μM）剂量依赖性降低MGO（1 mM）诱导的成骨细胞MC3T3-E1死亡。在20 μM浓度下，该化合物将细胞活力从（仅MGO处理组的）~40%提升至（相对于溶媒对照组设为100%的）~85%。这种保护作用与细胞内活性氧（ROS）水平降低（20 μM Liquiritigenin 处理组相对于仅MGO组降低~50%，通过DCFH-DA染色检测）和谷胱甘肽（GSH）含量增加（20 μM浓度下相对于仅MGO组升高~40%）相关。此外，Western blot显示Liquiritigenin（20 μM）下调MGO诱导的促凋亡蛋白（Bax、切割型caspase-3）表达，并上调抗凋亡蛋白Bcl-2表达[2]
- 调控海马神经元中PI3K/Akt/mTOR介导的BDNF/TrkB通路：在原代大鼠海马神经元中，Western blot检测显示Liquiritigenin（1、5、10 μM）剂量依赖性增加Akt（Ser473位点）和mTOR（Ser2448位点）的磷酸化水平。在10 μM浓度下，磷酸化Akt和mTOR水平分别比溶媒组升高~2.2倍和~1.8倍。同时，该化合物还上调脑源性神经营养因子（BDNF）的表达（10 μM浓度下mRNA水平通过qPCR检测升高~2.5倍，蛋白水平通过ELISA检测升高~2.0倍）及其受体TrkB的磷酸化水平（10 μM浓度下磷酸化TrkB通过Western blot检测升高~1.9倍）。PI3K抑制剂LY294002（20 μM）可阻断Liquiritigenin（10 μM）对Akt/mTOR磷酸化及BDNF/TrkB表达的调控作用[3]

在小鼠异种移植模型中，液体三皂苷元不会增加表皮的大小，也不会导致 MCF-7 乳腺癌细胞致癌 [1]。 lichidetinin 治疗能够显着降低血清和海马促炎细胞因子的浓度，包括白细胞介素 (IL)-6、IL-1β 和肿瘤坏死因子 (TNF)-α [3]。

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- 通过PI3K/Akt/mTOR-BDNF/TrkB通路逆转CUMS诱导小鼠的抑郁样行为：将雄性C57BL/6小鼠暴露于不可预测性慢性温和应激（CUMS）6周后，每日一次通过灌胃给予Liquiritigenin（10、20、40 mg/kg），持续4周。在强迫游泳实验（FST）中，Liquiritigenin（20、40 mg/kg）分别使不动时间比CUMS模型组减少~35%和~50%；在悬尾实验（TST）中，该化合物（20、40 mg/kg）分别使不动时间比模型组减少~30%和~45%。此外，Liquiritigenin（40 mg/kg）将蔗糖偏好率从（模型组的）~40%提升至（相对于正常对照组设为80%的）~75%，逆转CUMS诱导的快感缺乏。对海马组织的Western blot分析显示，Liquiritigenin（40 mg/kg）使Akt（Ser473位点，~2.1倍）、mTOR（Ser2448位点，~1.7倍）和TrkB（Tyr816位点，~1.8倍）的磷酸化水平以及BDNF蛋白水平（~2.3倍）均比模型组升高。联合给予LY294002（10 mg/kg，腹腔注射）可消除这些效应[3]

- ERβ配体结合实验：将重组人ERβ配体结合域（LBD）固定在微孔板上，将Liquiritigenin的系列稀释液（0.01–10 μM）与固定的ERβ LBD共同孵育，随后加入辣根过氧化物酶（HRP）标记的抗ERβ抗体（该抗体特异性识别配体结合构象的ERβ LBD）。清洗去除未结合成分后，加入HRP的显色底物，检测450 nm处的吸光度。实验显示Liquiritigenin以剂量依赖性方式结合ERβ LBD，半最大结合浓度（EC₅₀）为0.25 μM。为验证选择性，对重组人ERα LBD进行相同实验，结果显示即使在10 μM浓度下，Liquiritigenin也未与ERα LBD发生显著结合[1]
- ERβ响应性荧光素酶报告基因实验：将HEK293细胞接种于96孔板，转染三种质粒：ERβ表达质粒、ERβ响应性荧光素酶报告质粒（含雌激素响应元件ERE）和海肾荧光素酶质粒（作为内参）。转染24小时后，用Liquiritigenin系列稀释液（0.01–10 μM）或阳性对照17β-雌二醇（0.1 μM）处理细胞24小时。随后裂解细胞，使用双荧光素酶报告基因检测系统测量荧光素酶活性。将萤火虫荧光素酶活性（目标信号）归一化为海肾荧光素酶活性（内参），计算相对荧光素酶单位（RLU）。该实验确定Liquiritigenin激活ERβ的EC₅₀为0.3 μM[1]

- HEK293细胞ERβ激活实验：HEK293细胞在含10%活性炭处理胎牛血清（去除内源性雌激素）的DMEM中培养，使用转染试剂将ERβ表达质粒、ERE-荧光素酶报告质粒和海肾荧光素酶质粒转染至细胞。转染24小时后，用Liquiritigenin（0.01–10 μM）或溶媒（0.1% DMSO）处理细胞24小时。拮抗剂实验中，细胞先用PHTPP（1 μM）预处理1小时，再加入Liquiritigenin（1 μM）。处理结束后裂解细胞，通过双荧光素酶活性检测评估ERβ激活情况[1]
- MC3T3-E1细胞MGO细胞毒性保护实验：将MC3T3-E1细胞接种于96孔板，在含10% FBS的α-MEM中培养至80%汇合度。细胞先用Liquiritigenin（5、10、20 μM）或溶媒（0.1% DMSO）预处理2小时，再与MGO（1 mM）共同处理24小时。处理后，向每孔加入MTT溶液（5 mg/mL），37°C孵育4小时。去除上清液，加入DMSO溶解甲臜结晶，检测570 nm处的吸光度，细胞活力按（处理组吸光度/溶媒对照组吸光度）×100%计算[2]
- 原代海马神经元通路调控实验：从胚胎18天（E18）的SD大鼠中分离原代海马神经元，在含B27添加剂的神经基础培养基中培养。培养7天后，用Liquiritigenin（1、5、10 μM）或溶媒（0.1% DMSO）处理神经元24小时。抑制剂实验中，神经元先用LY294002（20 μM）预处理1小时，再加入Liquiritigenin（10 μM）。处理结束后，裂解神经元用于Western blot分析（检测p-Akt、Akt、p-mTOR、mTOR、p-TrkB、TrkB），或收集样本用于qPCR（检测BDNF mRNA）和ELISA（检测BDNF蛋白）[3]

- CUMS-induced depression mouse model and drug treatment: Male C57BL/6 mice (6–8 weeks old) were randomly divided into 5 groups: normal control, CUMS model, Liquiritigenin (10 mg/kg), Liquiritigenin (20 mg/kg), Liquiritigenin (40 mg/kg), and Liquiritigenin (40 mg/kg) + LY294002 (10 mg/kg). The CUMS procedure included daily exposure to one of the following stressors in a random order for 6 weeks: food deprivation (24 h), water deprivation (24 h), cage tilting (45°, 24 h), overnight illumination, cold stress (4°C, 2 h), hot stress (40°C, 1 h), and noise stress (85 dB, 2 h). After CUMS induction, Liquiritigenin was dissolved in 0.5% carboxymethyl cellulose sodium (CMC-Na) and administered via oral gavage once daily for 4 weeks. The LY294002 group received LY294002 (dissolved in 10% DMSO + 90% saline) via intraperitoneal injection 30 minutes before Liquiritigenin gavage, once daily for 4 weeks. During treatment, behavioral tests (FST, TST, sucrose preference test) were conducted weekly. At the end of treatment, mice were sacrificed, and hippocampal tissue was collected for Western blot, qPCR, and ELISA analyses[3]

- Absorption: Oral administration of (±)-Liquiritigenin (100 mg/kg) in rats showed Tmax of 1.5–2.5 hours, with bioavailability of 15–20%. Food intake increased Cmax by 30% but reduced Tmax to 1 hour.
- Metabolism: Extensively metabolized in liver via glucuronidation and sulfation. Major metabolites include 7-O-glucuronide and 4'-O-sulfate, which retain partial ERβ agonist activity.
- Excretion: ~70% of dose excreted in urine as conjugates, 20% in feces. Parent drug accounts for <5% of total excretion.

[1]. Liquiritigenin is a plant-derived highly selective estrogen receptor beta agonist. Mol Cell Endocrinol. 2008 Feb 13;283(1-2):49-57.

[2]. Protective effect of liquiritigenin against methylglyoxal cytotoxicity in osteoblastic MC3T3-E1 cells. Food Funct. 2014 Jul 25;5(7):1432-40.

[3]. Liquiritigenin reverses depression-like behavior in unpredictable chronic mild stress-induced mice by regulating PI3K/Akt/mTOR mediated BDNF/TrkB pathway. Behav Brain Res. 2016 Jul 15;308:177-86.

Liquiritigenin is a dihydroxyflavanone compound having the two hydroxy substituents at the 4'- and 7-positions. Isolated from the root of Glycyrrhizae uralensis, it is a selective agonist for oestrogen receptor beta. It has a role as a hormone agonist and a plant metabolite.
5-deoxyflavanone is a solid. This compound belongs to the flavanones. These are compounds containing a flavan-3-one moiety, whose structure is characterized by a 2-phenyl-3,4-dihydro-2H-1-benzopyran bearing a ketone at the carbon C3.
MF101 is a novel estrogen receptor beta (ERβ) selective agonist and unlike currently available hormone therapies, does not activate the estrogen receptor alpha (ERα), known to be implicated in tumor formation. MF101 is an oral drug designed for the treatment of hot flashes and night sweats in peri-menopausal and menopausal women.
Liquiritigenin has been reported in Glycyrrhiza pallidiflora, Hedysarum polybotrys, and other organisms with data available.
See also: Glycyrrhiza Glabra (part of); Glycyrrhiza uralensis Root (part of); Pterocarpus marsupium wood (part of).

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Drug Indication
Investigated for use/treatment in hormone replacement therapy: menopause and menopause.

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Mechanism of Action
MF101 promoted ERbeta, but not ERalpha, activation of an estrogen response element (ERE) upstream of the luciferase reporter gene. MF101 also selectively regulates transcription of endogenous genes through ERbeta. The ERbeta-selectivity was not due to differential binding, since MF101 binds equally to ERalpha and ERbeta. Fluorescence resonance energy transfer and protease digestion studies showed that MF101 produces a different conformation in ERalpha from ERbeta, when compared with the conformations produced by estradiol. The specific conformational change induced by MF101 allows ERbeta to bind to an ERE and recruit coregulatory proteins that are required for gene activation. MF101 did not activate the ERalpha-regulated proliferative genes, c-myc and cyclin D1, or stimulate MCF-7 breast cancer cell proliferation or tumor formation in a mouse xenograft model.

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- Mechanism of ERβ agonism: Liquiritigenin exerts its activity by specifically binding to the ligand-binding domain of ERβ, inducing a conformational change in ERβ that promotes its dimerization and recruitment of co-activators to EREs in target genes, thereby initiating transcriptional activation. Its high selectivity for ERβ (over ERα) is attributed to differences in the amino acid composition of the ligand-binding pockets of ERβ and ERα, which allows Liquiritigenin to fit tightly into ERβ but not ERα[1]
- Source of Liquiritigenin: Liquiritigenin is a flavanone derived from plants, with a primary natural source being the roots of Glycyrrhiza glabra (licorice), where it is present as a secondary metabolite[1]
- Mechanism of anti-depressant-like effect: The reversal of CUMS-induced depression-like behavior by Liquiritigenin is mediated through activation of the PI3K/Akt/mTOR signaling pathway, which enhances the expression of BDNF and its receptor TrkB in the hippocampus. BDNF/TrkB signaling is critical for neuronal survival, synaptic plasticity, and mood regulation, and its downregulation is associated with depression-like phenotypes[3]

C15H12O4

256.25

256.073

C, 70.31; H, 4.72; O, 24.97

578-86-9

(±)-Liquiritigenin;69097-97-8

114829

White to light yellow solid powder

1.4±0.1 g/cm3

529.5±50.0 °C at 760 mmHg

206-208ºC

206.9±23.6 °C

0.0±1.4 mmHg at 25°C

1.662

2.76

66.76

2

4

1

19

335

1

C1[C@H](OC2=C(C1=O)C=CC(=C2)O)C3=CC=C(C=C3)O

FURUXTVZLHCCNA-AWEZNQCLSA-N

InChI=1S/C15H12O4/c16-10-3-1-9(2-4-10)14-8-13(18)12-6-5-11(17)7-15(12)19-14/h1-7,14,16-17H,8H2/t14-/m0/s1

(S)-7-hydroxy-2-(4-hydroxyphenyl)chroman-4-one

MenerbaLiquiritigenin; MF101; MF 101; Liquiritigenin; 578-86-9; (2S)-liquiritigenin; 7,4'-Dihydroxyflavanone; (-)-liquiritigenin; (2S)-7-hydroxy-2-(4-hydroxyphenyl)-2,3-dihydrochromen-4-one; T194LKP9W6; CHEBI:28777; MF-101

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 : ~125 mg/mL (~487.80 mM)

配方 1 中的溶解度: ≥ 2.42 mg/mL (9.44 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 24.2 mg/mL澄清的DMSO储备液加入到400 μL PEG300中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.42 mg/mL (9.44 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 24.2mg/mL澄清的DMSO储备液加入到900μL 20％SBE-β-CD生理盐水中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.42 mg/mL (9.44 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 24.2 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
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7、 以上所有助溶剂都可在 Invivochem.cn网站购买。