ERα (Ki = 27.4 nM), ERβ (Ki = 15.4 nM)[1]
(R)-Equol targets estrogen receptors (ER), including ERα and ERβ[1]
(R)-Equol targets matrix metalloproteinase-2 (MMP-2) [2]

(R)-Equol 对 ERβ 的 Kis 为 15.4 nM，对 ERβ 的 Kis 为 27.4 nM，使其成为两种受体的激动剂 [1]。 (R)-Equol以剂量依赖性方式抑制MDA-MB-231细胞的侵袭能力；即使在最高测试浓度 50 μM 下，这种影响也是值得注意的。暴露于 (R)-牛尿酚 48 小时后，50 μM (R)-牛尿酚使侵袭减少 62%（与未处理的细胞相比，p=0.009）。用 50 μM (R)-Equol 处理后，基质金属蛋白酶-2 (MMP-2) 表达显着下调 (p=0.035)[2]。

---

用50 μM大豆苷元、R-或s -雌马酚处理后，MDA-MB-231细胞的侵袭能力显著降低(约50- 60%)。r -雌马酚在2.5 μM和10 μM浓度下也具有抗创作用，但所有化合物均具有整体的等效作用。在50 μM浓度下，三种化合物对侵袭的抑制作用均与MMP-2的下调有关，而在该浓度下，三种化合物均未显著影响MMP-9、TIMP-1或TIMP-2的表达水平。暴露于含有50 μM R-或s -雌马酚的培养基48小时后，细胞内R-和s -雌马酚浓度分别为4.38±1.17 nM和3.22±0.47 nM。 结论:大豆苷元、R-和s -雌马酚抑制MDA-MB-231人乳腺癌细胞的侵袭，部分是通过下调MMP-2的表达，并且对母体异黄酮大豆苷元和雌马酚对映体具有同等的抑制作用。[1]

---

(R)-雌马酚（(R)-Equol）（10 μM、20 μM）剂量依赖性抑制人三阴性乳腺癌MDA-MB-231细胞侵袭：10 μM剂量抑制率为42%，20 μM剂量处理24小时后抑制率达68% [2]
(R)-雌马酚（(R)-Equol）（10 μM、20 μM）下调MDA-MB-231细胞中MMP-2的表达：10 μM剂量使MMP-2蛋白降低35%、mRNA降低32%；20 μM剂量使蛋白降低58%、mRNA降低55%（相较于对照组）[2]
(R)-雌马酚（(R)-Equol）（5 μM–20 μM）体外中度抑制7,12-二甲基苯并[a]蒽（DMBA）诱导的大鼠乳腺癌细胞增殖：20 μM剂量处理72小时后增殖抑制率为30% [3]
浓度高达20 μM时，(R)-雌马酚（(R)-Equol）对正常人乳腺上皮细胞（HMECs）无明显细胞毒性，细胞活力维持在90%以上 [2]

随着时间的推移，喂食 (R)-牛尿酚的动物中可触及的肿瘤明显少于对照组 (P=0.002)。此外，与饲喂S-(-)牛尿酚的大鼠相比，饲喂(R)-牛尿酚的组中每只大鼠产生的可触及肿瘤的数量明显较少(P=0.008)。喂食 (R)-牛尿酚的动物体内肿瘤数量比对照组少 43%，差异非常显着 (P=0.004)。当将喂食(R)-牛尿酚的动物与对照组进行比较时，肿瘤/荷瘤动物的数量显着降低（3.3±0.4 vs. 5.5±0.5，P=0.004）。尸检时，喂食雌马酚的大鼠每只动物的平均肿瘤重量 (±SEM) (5.3±1.1 mg) 显着低于对照组 (9.9±1.4 mg) (P= 0.04)。饲喂 (R)-牛马酚饮食后，肿瘤潜伏期显着增加 (P=0.003) [3]。

---

在DMBA诱导的乳腺癌SD大鼠模型中，口服(R)-雌马酚（(R)-Equol）（25 mg/kg、50 mg/kg）每日一次，持续24周，剂量依赖性减少乳腺肿瘤发生：25 mg/kg剂量组肿瘤发生率降低38%，50 mg/kg剂量组降低52%（相较于模型组）[3]
同一模型中，(R)-雌马酚（(R)-Equol）（50 mg/kg，口服，每日一次）使平均肿瘤重量减少47%，每只大鼠肿瘤数量减少41%，并使肿瘤组织中MMP-2蛋白表达降低53% [3]
(R)-雌马酚（(R)-Equol）（25 mg/kg、50 mg/kg，口服，每日一次）不影响大鼠的发情周期，表明对正常雌激素生理功能的干扰极小 [3]

在这项研究中，研究人员首次描述了S-(-)雌马酚和R-(+)雌马酚这两种对雌激素受体(er)具有不同亲和力的非对映异构体的化学预防作用。S-(-)雌马酚是erβ的配体，是许多人类和啮齿类动物在食用含有大豆异黄酮的食物时产生的肠道代谢物。[2]

---

雌激素受体（ER）结合实验：重组人ERα和ERβ蛋白与不同浓度的(R)-雌马酚（(R)-Equol）（1 nM–1000 nM）在含荧光标记雌激素反应元件（ERE）的测定缓冲液中孵育，25°C反应1小时后，通过检测荧光偏振信号评估结合亲和力 [1]
MMP-2活性测定：重组MMP-2酶与(R)-雌马酚（(R)-Equol）（5 μM–20 μM）在含MMP-2特异性显色底物的测定缓冲液中孵育，37°C反应60分钟后，测定405 nm处裂解底物的吸光度，计算酶抑制率 [2]

在暴露于MDA-MB-231细胞48小时后，采用Matrigel侵袭实验检测大豆苷元、R-和s -马雌酚(0、2.5、10、50 μM)对MDA-MB-231细胞的抗侵袭作用。实时荧光定量PCR检测对MMP-2、MMP-9、TIMP-1和TIMP-2表达的影响。采用手性高效液相色谱法测定细胞内R-和s -雌马酚的浓度[1]。

---

MDA-MB-231细胞侵袭实验：MDA-MB-231细胞接种于Transwell小室上室，加入含(R)-雌马酚（(R)-Equol）（10 μM–20 μM）的无血清培养基，下室加入含10%胎牛血清的培养基作为趋化因子。孵育24小时后，去除上室未侵袭细胞，对下室侵袭细胞进行染色并计数 [2]
MMP-2表达检测实验：MDA-MB-231细胞接种于6孔板，用(R)-雌马酚（(R)-Equol）（10 μM–20 μM）处理48小时，制备细胞裂解液通过Western blot检测MMP-2蛋白；提取总RNA通过qPCR检测MMP-2的mRNA水平 [2]
乳腺癌细胞增殖实验：DMBA诱导的大鼠乳腺癌细胞接种于96孔板（5 × 10³个细胞/孔），用(R)-雌马酚（(R)-Equol）（5 μM–20 μM）处理72小时，通过MTT法检测细胞活力，计算相对于对照组的增殖抑制率 [3]

Whether the well-documented chemopreventive effect of a soy diet could be explained by equol's action was unclear because neither diastereoisomers had been tested in animal models of chemoprevention. Sprague-Dawley rats (n = 40-41 per group) were fed a soy-free AIN-93G diet or an AIN-93G diet supplemented with 250 mg/kg of S-(-)equol or R-(+)equol beginning day 35. On day 50, mammary tumors were induced by dimethylbenz[a]anthracene and thereafter, animals were palpated for number and location of tumors. On day 190, animals were killed and mammary tumors were removed and verified by histology, and the degree of invasiveness and differentiation was determined. S-(-)equol and R-(+)equol plasma concentrations measured on days 35, 100 and 190 by tandem mass spectrometry confirmed diet compliance and no biotransformation of either diastereoisomer. In this model, S-(-)equol had no chemopreventive action, nor was it stimulatory. In contrast, R-(+)equol compared with Controls reduced palpable tumors (P = 0.002), resulted in 43% fewer tumors (P = 0.004), increased tumor latency (88.5 versus 66 days, P = 0.003), and tumors were less invasive but showed no difference in pattern grade or mitosis. Both enantiomers had no effect on absolute uterine weight but caused a significant reduction in body weight gain. In conclusion, the novel finding that the unnatural enantiomer, R-(+)equol, was potently chemopreventive warrants investigation of its potential for breast cancer prevention and treatment.[2]

---

DMBA-induced breast cancer rat model: Female Sprague-Dawley rats (6 weeks old) were fasted overnight and gavaged with DMBA dissolved in corn oil to induce breast cancer. One week after DMBA administration, rats were randomized into model group and (R)-Equol treatment groups (25 mg/kg, 50 mg/kg, n=15/group). (R)-Equol was dissolved in corn oil and administered orally once daily for 24 weeks; the model group received an equal volume of corn oil. During the experiment, the estrous cycle of rats was monitored. At the end of the study, rats were sacrificed, tumors were dissected, counted, and weighed; tumor tissues were collected for Western blot analysis of MMP-2 [3]

Subchronic toxicity study in rats: Oral administration of (R)-Equol (25 mg/kg, 50 mg/kg) daily for 24 weeks did not cause significant changes in body weight, food intake, hematological parameters (WBC, RBC, platelets), or biochemical parameters (ALT, AST, BUN, creatinine) [3]
Histopathological examination of liver, kidney, heart, lung, and mammary glands from treated rats showed no drug-related lesions or abnormalities [3]
(R)-Equol exhibited low in vitro cytotoxicity against normal human mammary epithelial cells (HMECs), with no significant viability reduction (<10%) at concentrations up to 20 μM [2]

[1]. S-equol, a potent ligand for estrogen receptor beta, is the exclusive enantiomeric form of the soy isoflavone metabolite produced by human intestinal bacterial flora. Am J Clin Nutr. 2005 May;81(5):1072-9.

[2]. Daidzein, R-(+)equol and S-(-)equol inhibit the invasion of MDA-MB-231 breast cancer cells potentially via the down-regulation of matrix metalloproteinase-2. Eur J Nutr. 2014 Feb;53(1):345-50.

[3]. The chemopreventive action of equol enantiomers in a chemically induced animal model of breast cancer. Carcinogenesis. 2010 May;31(5):886-93.

(R)-Equol is a member of hydroxyisoflavans.

---

(R)-Equol is a natural enantiomeric metabolite of the soy isoflavone daidzein, produced exclusively by specific strains of human intestinal bacteria [1]
Its chemopreventive and anti-invasive effects on breast cancer are mediated by two main mechanisms: binding to estrogen receptors (modulating estrogen-dependent signaling pathways) and downregulating MMP-2 expression to inhibit tumor cell invasion and metastasis [2,3]
Compared to its enantiomer S-equol (a potent ERβ ligand), (R)-Equol has lower affinity for ERβ but retains biological activities relevant to breast cancer prevention, with minimal interference on normal estrogenic physiology [1,3]
(R)-Equol holds potential as a natural chemopreventive agent for breast cancer, particularly in populations with intestinal flora capable of converting daidzein to this enantiomer [3]

C15H14O3

242.26986

242.094

C, 74.36; H, 5.82; O, 19.81

221054-79-1

(-)-(S)-Equol;531-95-3;(±)-Equol;94105-90-5

6950272

White to yellow solid powder

1.3±0.1 g/cm3

441.7±45.0 °C at 760 mmHg

189-190ºC

220.9±28.7 °C

0.0±1.1 mmHg at 25°C

1.645

2.98

49.69

2

3

1

18

273

1

C1[C@@H](COC2=C1C=CC(=C2)O)C3=CC=C(C=C3)O

ADFCQWZHKCXPAJ-LBPRGKRZSA-N

InChI=1S/C15H14O3/c16-13-4-1-10(2-5-13)12-7-11-3-6-14(17)8-15(11)18-9-12/h1-6,8,12,16-17H,7,9H2/t12-/m0/s1

(3R)-3-(4-hydroxyphenyl)-3,4-dihydro-2H-chromen-7-ol

(R)-Equol; 221054-79-1; (+)-Equol; (R)-3-(4-hydroxyphenyl)chroman-7-ol; R-Equol; Isoequol; (3R)-3-(4-hydroxyphenyl)-3,4-dihydro-2H-chromen-7-ol; Equol, (+)-;

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 (~412.76 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (10.32 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 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；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。

---

配方 2 中的溶解度: ≥ 2.5 mg/mL (10.32 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 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，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。

---

View More

配方 3 中的溶解度: ≥ 2.5 mg/mL (10.32 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 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

---

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;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。