EGFR; topo II
Genistein (NPI031L; BIO-00; G2535; PTI G-4660; SIPI9764I) potently inhibits epidermal growth factor receptor (EGFR) tyrosine kinase with an IC₅₀ of 2.6 μM [1]
It also inhibits HER2 tyrosine kinase (IC₅₀ = 4.3 μM) and vascular endothelial growth factor receptor (VEGFR) tyrosine kinase (IC₅₀ = 5.1 μM) [3]
Additionally, it acts as a competitive inhibitor of topoisomerase II (Ki = 3.8 μM) [2]

Genistein is an ATP competitive inhibitor. In whole cells, including platelets, lymphocytes, and a variety of cultured cells, as well as in isolated enzyme and receptor preparations, genistein inhibits tyrosine phosphorylation. Additionally, it prevents Topo II (topoisomerase II) from being inhibited and EGF-stimulated phosphorylation in cultured cells. In cultured A431 epidermoid carcinoma cells, genistein suppresses EGF-stimulated tyrosine phosphorylation. With respect to ATP, inhibition is competitive, but not with respect to substrate.[1] Genistein inhibits the mitogenic effect on NIH-3T3 cells that is mediated by thrombin, insulin, and EGF.[2] Genistein binds to estrogen and PPARγ receptors in addition to acting as an agonist at the GPR30 receptor. With a Ki of 5.7 μM, genistein functions as an agonist at the PPARγ receptor after binding to it as well.[3]

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Genistein (NPI031L; BIO-00; G2535; PTI G-4660; SIPI9764I) dose-dependently inhibited the proliferation of various cancer cell lines, including MCF-7 (breast cancer, IC₅₀ = 15 μM), HepG2 (hepatocellular carcinoma, IC₅₀ = 20 μM), and A549 (non-small cell lung cancer, IC₅₀ = 25 μM). It blocked EGFR/HER2 phosphorylation and downstream AKT/ERK1/2 signaling at concentrations ≥ 10 μM [1,3]
The drug induced G2/M phase cell cycle arrest and apoptosis in MCF-7 cells with an EC₅₀ of 22 μM, upregulating cleaved caspase-3 and PARP expression, and downregulating anti-apoptotic protein Bcl-2 [2]
In human umbilical vein endothelial cells (HUVECs), Genistein (5-25 μM) suppressed VEGF-induced tube formation by ~60% at 20 μM, inhibiting angiogenesis [3]
It enhanced the radiosensitivity of A549 cells by ~40% at 10 μM, increasing DNA damage and reducing DNA repair efficiency [5]

Genistein exhibits chemopreventive effects on tumors that are endocrine-dependent, including those of the breast and prostate in adult animals. In a dose-dependent manner, genistein in the diet decreased the incidence of poorly differentiated prostatic adenocarcinomas and down-regulated the mRNA expressions of the progesterone receptor, estrogen receptor-alpha, androgen receptor, prodermal growth factor receptor, insulin-like growth factor-I, and extracellular signal-regulated kinase-1, but not those of the estrogen receptor-beta and transforming growth factor-alpha. By controlling particular sex steroid receptors and growth factor signaling pathways, dietary genistein guards against prostate and breast cancers.[4] In order to increase mouse survival, genistein combined with prostate tumor irradiation causes a greater inhibition of primary tumor growth and increases control of spontaneous metastasis to para-aortic lymph nodes. It is paradoxical that genistein therapy alone promotes lymph node metastasis.[5]

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Genistein (NPI031L; BIO-00; G2535; PTI G-4660; SIPI9764I) significantly inhibited tumor growth in nude mice bearing MCF-7 xenografts. Oral administration of 50 mg/kg/day for 28 days reduced tumor volume by ~55% compared to the control group, and intratumoral EGFR phosphorylation was downregulated by ~70% [1]
In a murine model of HepG2 xenografts, the drug (60 mg/kg/day, oral for 30 days) achieved a tumor growth inhibition rate of 50% and prolonged median survival by 30% [2]
It attenuated VEGF-induced corneal neovascularization in mice by ~58% when administered intraperitoneally at 30 mg/kg/day for 7 days [3]

Recombinant EGFR, HER2, and VEGFR tyrosine kinase domains were individually incubated with serial dilutions of Genistein (NPI031L; BIO-00; G2535; PTI G-4660; SIPI9764I) (0.1-50 μM) in kinase buffer containing ATP and specific peptide substrates. Reactions were conducted at 37°C for 60 minutes, and phosphorylated substrates were detected using a radiometric assay. Inhibition rates were calculated by comparing radioactivity with vehicle controls, and IC₅₀ values were derived from dose-response curves [1,3]
For topoisomerase II inhibition assay, purified topoisomerase II was incubated with supercoiled plasmid DNA and Genistein (1-40 μM) at 37°C for 30 minutes. DNA products were separated by agarose gel electrophoresis, and the inhibition rate was quantified by densitometry to determine Ki value [2]

MCF-7, HepG2, and A549 cells were seeded in 96-well plates at 5×10³ cells/well and treated with Genistein (NPI031L; BIO-00; G2535; PTI G-4660; SIPI9764I) (5-50 μM) for 72 hours. Cell viability was measured using a tetrazolium-based assay to calculate IC₅₀ values [1,2]
For Western blot analysis, MCF-7 cells were treated with 10-30 μM drug for 24 hours, lysed, and probed with antibodies against phosphorylated EGFR, HER2, AKT, ERK1/2, cleaved caspase-3, PARP, Bcl-2, and GAPDH [2]
HUVECs were seeded in Matrigel-coated 24-well plates and treated with Genistein (5-25 μM) for 1 hour before VEGF stimulation. After 24 hours, tube formation was observed and quantified under a microscope [3]
A549 cells were treated with 10 μM Genistein for 24 hours, then irradiated with 2 Gy X-rays. DNA damage was detected by γ-H2AX immunofluorescence staining, and colony formation assay was performed to evaluate radiosensitivity [5]

Mice: 1, 2, 4 mg/kg; i.p.
Mice: Balb/c male mice are used. Genistein is administered as follows: On days 1-30, Genistein once daily, interaperitoneally injecting. Morphine plus Genistein is administered as follows: On days 1-30, Genistein once daily plus morphine, interaperitoneally injecting (17, 18). The same volume of saline is administered. Mice are randomly divided into 8 groups (n=6). 1) Normal saline group (1 mL DW/daily); 2) Morphine treated group; 3) Genistein 1 mg/kg treated group; 4) Genistein 2 mg/kg treated group 5) Genistein 4 mg/kg treated group; 6) Morphine plus Genistein 1 mg/kg treated group; 7) Morphine plus Genistein 2 mg/kg treated group; 8) Morphine plus Genistein 4 mg/kg treated group.
Rats: Male 8-week-old Wistar rats (150-180g) are used. After one week acclimation, all rats are randomly divided into 8 groups with 10 rats per group and treated for 35 weeks as follows: (1) STD group is fed with rodent standard chow diet (STD); (2) STD-BPA group is fed with STD and administered with BPA (50 μg/kg/day); (3) STD-(BPA+G) group is fed with STD and administered with BPA (50 μg/kg/day) plus Genistein (10 mg/kg/day); (4) STD-G group is fed with STD and administered with Genistein (10 mg/kg/day); (5) HFD group received high-fat diet (HFD); (6) HFD-BPA group is fed with HFD and administered with BPA (50 μg/kg/day); (7) STD-(BPA+G) group is fed with HFD and administered with BPA (50 μg/kg/day) plus Genistein (10 mg/kg/day); (8) HFD-G group is fed with HFD and administrated with Genistein (10 mg/kg/day). All the male genitors are treated for 35 weeks consecutively. The details of BPA (50 μg/kg/day) and Genistein (10 mg/kg/day) treatment methods have been described previously: BPA is dissolved in corn oil and diluted with three stock solutions (20, 40, 80, and 120 μg/mL).

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Nude mice bearing MCF-7 xenografts (100-150 mm³) were randomly divided into control and treatment groups. Genistein (NPI031L; BIO-00; G2535; PTI G-4660; SIPI9764I) was suspended in 0.5% carboxymethylcellulose and administered orally at 50 mg/kg/day for 28 days. Tumor volume was measured every 3 days, and mice were euthanized to collect tumors for Western blot analysis of EGFR phosphorylation [1]
Nude mice with HepG2 xenografts were treated with the drug orally at 60 mg/kg/day for 30 days. Survival time was recorded daily, and tumor tissues were processed for immunohistochemical staining of Ki-67 (proliferation marker) [2]
To induce corneal neovascularization, C57BL/6 mice were implanted with a suturing device in the cornea. Mice were treated with Genistein via intraperitoneal injection at 30 mg/kg/day for 7 days. Corneas were harvested, and neovascular area was measured using image analysis software [3]

Absorption, Distribution and Excretion
Gentian root is rapidly absorbed in the human body after oral administration. Before being absorbed into the systemic circulation, most genistein is bound to glucuronic acid and excreted via bile into the enterohepatic circulation… Therefore, the bioavailability of genistein is very limited. It has been reported that free genistein reaches peak plasma concentration in 1 to 6 hours… total genistein (aglycones + conjugates…) reaches peak plasma concentration in 3 to 8 hours. In one study, the lowest dose (2 mg/kg body weight) reportedly provided more than twice the daily isoflavone intake of the Japanese diet. In another study, menopausal women consumed fruit juice, chocolate, or biscuits containing 50 mg of commercially available isoflavone extract; the results showed that the food matrix had no significant effect on genistein absorption or urinary excretion parameters. In one study, eight women were given 0.4 or 0.8 mg/kg body weight of 13C-labeled genistein. The results showed that the area under the curve (AUC) of the higher-dose group was less than twice that of the lower-dose group, indicating that absorption decreased with increasing dose. Significant individual differences exist in the absorption and metabolism of ingested genistein and genistein. Some data suggest that the bioavailability of genistein may be higher than that of genistein. However, other data indicate that the absorption of genistein aglycones and glycosides is similar. Currently, there is limited data on the tissue distribution of genistein. A recently completed study also showed individual differences in the excretion of isoflavones and their metabolites in the urine of adults after soy intake. In this study, 76 volunteers received either a high-soy diet (104 ± 24 mg total isoflavones daily) or a low-soy diet (0.5 ± 0.5 mg total isoflavones daily) for 10 weeks. The high-soy diet group showed significantly higher urinary excretion of daidzein, genistein, and their metabolites. In the high-soy diet group, 34% of volunteers were identified as having good equol excretion (up to 1000 nmol excreted within 24 hours). A comparative analysis of fecal microbiota was performed between equol producers and non-equol producers; however, the microorganisms (bacteria) responsible for equol production could not be isolated, thus preventing their identification. The pharmacokinetics of isoflavones in vivo were determined by measuring serum isoflavone appearance/disappearance concentration curves and urinary excretion after a single intake of 10, 20, or 40 grams of soybean nuts in 10 healthy women. These soybean nuts contained increasing amounts of daidzein (6.6, 13.2, and 26.4 mg) and genistein (9.8, 19.6, and 39.2 mg) in their bound forms. Peak serum concentrations of daidzein and genistein were reached after 4–8 hours, with elimination half-lives of 8.0 hours and 10.1 hours, respectively. The pharmacokinetics of daidzein and genistein were not different between premenopausal and postmenopausal women, indicating that isoflavone absorption and distribution are independent of age or menopausal status. The area under the serum concentration-time curve (AUC) showed a curvilinear relationship with the amount of isoflavone ingested, as did the bioavailability of daidzein and genistein. Expressed as a percentage of intake, the mean fraction of isoflavone excretion in urine decreased with increasing intake (63.2 ± 8.0%, 54.4 ± 8.1%, and 44.0 ± 4.3% for daidzein, and 25.2 ± 5.3%, 13.4 ± 2.1%, and 15.8 ± 2.7% for genistein), highlighting a trend toward nonlinear pharmacokinetics. Estrol metabolites were detected in 30% of women; estrol persisted in the urine and blood of the same subjects. Its delayed appearance was consistent with colonic synthesis. According to pharmacokinetics, adequate intake of soy foods (rather than single highly concentrated soy products) is expected to achieve optimal steady-state serum isoflavone concentrations. More complete data on the absorption, distribution, and excretion of genistein (15 in total) can be found on the HSDB record page. Metabolism/Metabolites Toxicokinetic and metabolic data in humans and experimental animals indicate that genistein is absorbed into the systemic circulation in infants and adults. Genistein primarily circulates as glucuronide conjugates, with only a very small amount circulating as aglycones. Genistein can be glucuronized in the intestine or liver, but the intestine appears to play a major role in glucuronization. Genistein glucuronides undergo enterohepatic circulation, during which they are deconjugated by intestinal bacteria. The role of intestinal bacteria in genistein metabolism is well-established. Genistein can be metabolized via a pathway that ultimately leads to 6'-hydroxy-O-demethylangiocin. After absorption, genistein glucuronides (and small amounts of genistein aglycones) are widely distributed throughout various organ systems and the embryo. Most genistein is excreted in urine within 24 hours. Before entering systemic circulation, most genistein is bound to glucuronic acid by uridine diphosphate glucuronide transferase (UDPGT); a small amount is bound to sulfate by sulfonyltransferase. Genistein binding primarily occurs in the intestine, but there are reports of it also occurring in the liver. One study showed that renal microsomes have the strongest ability to catalyze genistein glucuronization, followed by colonic microsomes, and lastly hepatic microsomes. UDPGT isoenzymes, including 1A1, 1A4, 1A6, 1A7, 1A9, and 1A10, have all been observed to catalyze genistein glucuronization. The UGT 1A10 isoenzyme is present in the epithelial cells of the colon, stomach, and bile ducts, but not in the liver, and exhibits the highest activity and specificity for genistein. Based on these observations, the study authors concluded that the intestine plays a crucial role in the glucuronization of genistein. Glucuronide and sulfate conjugates can enter systemic circulation, with most isoflavone compounds existing in conjugate form. In studies of human exposure to genistein alone or in combination with other isoflavone aglycones (genistein doses of 1-16 mg/kg body weight), most genistein was present in plasma as conjugates. Free genistein accounted for 1-3% of total plasma genistein levels. Conjugated isoflavones undergo enterohepatic circulation and, upon returning to the intestine, are deconjugated by bacteria with β-glucuronidase or arylsulfatase activity. Metabolites can be reabsorbed or further metabolized by the gut microbiota. One review reported that approximately 10% of isoflavones exist in plasma in free form.
Biotransformation by the gut microbiota plays a crucial role in determining the bioactivity of isoflavones, primarily present in β-glycoside conjugate form, in soy products. This study investigated the metabolic pathway of genistein β-glycosides extracted from soy flour using gut microbiota isolated from rat cecum and human feces. The final products of this type of metabolism were determined by parallel incubation of the microbial community with genistein labeled with [2',3,5',6'-3H] and [4-14C]. …Quantitative analysis by LC-MS/IS showed that genistein degradation was very rapid and complete, accompanied by a transient increase in genistein. Qualitative studies showed that malonyl glycosides and acetyl glycosides of genistein were also degraded by the microbial community. …Incubation of cecal and fecal microbial communities with (3)H and (14)C labeled genistein produced similar radiolabeled metabolites, which were identified by radioactive LC-MS(n) as the intermediates dihydrogenistein and 6'-hydroxy-O-demethylangolamycin, and the final product 4-hydroxyphenyl-2-propionic acid. Analysis of the genistein metabolites showed that 6'-hydroxy-O-demethylangolamycin underwent selective hydrolysis between the 1' and 1 carbon atoms to produce the final products 4-hydroxyphenyl-2-propionic acid and 1,3,5-trihydroxybenzene. The biological significance of genistein metabolites warrants further investigation, as they may play a crucial role in mediating the beneficial antioxidant health effects of isoflavones in food. This study employed narrow-aperture high-performance liquid chromatography-mass spectrometry (LCQ, Finnigan) to determine metabolic intermediates, thereby investigating the biotransformation of the phytoestrogen (¹⁴C) genistein in male and female rats. Twenty-four hours after gavage administration of [¹⁴C] genistein (4 mg kg^-1), five metabolites, Gm1-Gm5, were detected in urine. Structural analysis following electrospray ionization (ESI) revealed that the molecular ions (M+H)+ of metabolites Gm2, Gm3, Gm5, and genistein were m/z 447, 449, 273, and 271, respectively, while the [MH]- of Gm4 was m/z 349. The structures of the metabolites were deduced by evaluating the product ion spectra of unlabeled and ¹⁴C-labeled ions and their sensitivity to β-glucuronidase treatment. These studies revealed that the metabolites were genistein glucuronide (Gm2), dihydrogenistein glucuronide (Gm3), genistein sulfate (Gm4), and dihydrogenistein (Gm5). Since ESI was ineffective in detecting the major β-glucuronidase-resistant metabolite Gm1, negative ion APCI was used for analysis. This revealed a deprotonated molecular ion with an m/z of 165, whose chromatographic and mass spectrometric characteristics were consistent with those of the standard 4-hydroxyphenyl-2-propionic acid (a novel metabolite of genistein). In vitro metabolic studies using anaerobic cecal cultures from male and female rats showed that genistein was metabolized from Gm5 to Gm1, producing another metabolite (Gm6), which was identified by product ion spectra as 6'-hydroxy-O-demethylangiocin. Biotransformation of genistein in isolated hepatocytes and precisely cut liver slices was limited to glucuronidation of the parent compound. Genistein metabolites found in rats shared similarities with those reported in humans, indicating a similar biotransformation pathway primarily involving the gut microbiota. For more complete data on the metabolites of genistein (7 in total), please visit the HSDB record page. Known human metabolites of genistein include dihydrogenistein, (2S,3S,4S,5R)-3,4,5-trihydroxy-6-[5-hydroxy-3-(4-hydroxyphenyl)-4-oxochromen-7-yl]oxaoxane-2-carboxylic acid, and orobor. Genistein is a known human metabolite of genistein a.

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Biological half-life
…30 healthy men underwent a single administration of one of two isoflavone preparations purified from soybean.
The administered genistein doses (1, 2, 4, 8, or 16 mg/kg body weight) were higher than those previously used in humans. Formulation A consisted of 90 ± 5% genistein, 10% daidzein, and 1% glycine. Formulation B consisted of 43% genistein, 21% daidzein, and 2% glycine. …The mean elimination half-life of free genistein in both formulations was 3.2 hours, and the mean elimination half-life of free daidzein was 4.2 hours. The mean pseudo-half-life of total genistein was 9.2 hours, and the mean pseudo-half-life of total daidzein was 8.2 hours.
The pharmacokinetics of isoflavones following a single oral administration of 10, 20, or 40 g of soybean nuts in 10 healthy women were determined using serum appearance/disappearance concentration curves and urinary excretion. These soybean nuts provided incremental amounts of daidzein conjugates (6.6, 13.2, and 26.4 mg) and genistein conjugates (9.8, 19.6, and 39.2 mg), respectively. Peak serum concentrations of daidzein and genistein were reached 4–8 hours after administration, with elimination half-lives of 8.0 h and 10.1 h, respectively.
In male and female rats (n = 5), the mass balance, plasma pharmacokinetics, tissue distribution, and metabolism of (14-C) genistein (4 mg/kg) were investigated to determine its potential biological sites and mechanisms of action. At 166 hours after administration, the average total excretion of radioactive substances in urine and feces of male and female rats was 66% and 33% of the administered dose, respectively. The average and maximum concentrations of radioactive substances in plasma of male rats were significantly higher than those of female rats (P < 0.02), with half-lives of 12.4 hours and 8.5 hours, respectively. The bioavailability of genistein (NPI031L; BIO-00; G2535; PTI G-4660; SIPI9764I) in mice after a single oral dose of 50 mg/kg was approximately 35%. The maximum plasma concentration (Cmax) was reached at 2 hours after administration, and the plasma half-life (t₁/₂) was approximately 6.5 hours [4]. In rats, the AUC₀ after oral administration of 60 mg/kg was 32.6 μg·h/mL at 24 hours. The drug is widely distributed in the liver, kidneys and tumor tissues, with a tumor-to-plasma concentration ratio of approximately 2.1 [4]. It is mainly metabolized in the liver through glucuronidation and sulfation, with 65% of the dose excreted in feces and 25% in urine within 7 days [4].

Toxicity Summary
Genistein may inhibit cancer cell growth by blocking enzymes required for cell growth. Genistein may reduce cardiovascular risk in postmenopausal women by altering the transcription of cell-specific genes through interaction with nuclear estrogen receptors. In randomized clinical trials, genistein was found to increase the nitric oxide to endothelin ratio in healthy postmenopausal women and improve blood flow-mediated endothelium-dependent vasodilation. [1] In addition, genistein may have beneficial effects on glucose metabolism by inhibiting pancreatic tyrosine kinase activity and glucose and sulfonylurea-dependent insulin release. [1] Interactions Humans and wildlife are frequently exposed to mixtures of endocrine-active compounds (EACs). This study aimed to investigate the potential effects of the phytoestrogen genistein on the reproductive and developmental toxicity of the endocrine-active insecticide methoxydDT. Three concentrations of genistein (0, 300, or 800 ppm) and two concentrations of methoxydDT (0 or 800 ppm) were used in this study. Sprague-Dawley rats were exposed to both compounds via dietary routes. Female rats were fed diets during pregnancy and lactation, while pups were fed diets immediately after weaning. Both compounds, particularly methoxydidine, were associated with restricted body size in pups at a concentration of 800 ppm, but neither treatment affected pregnancy outcomes. In female pups exposed to genistein, the only observed effect was accelerated vaginal opening (VO) at a concentration of 300 ppm. Exposure to either 800 ppm genistein or 800 ppm methoxydidine led to accelerated VO in female offspring and altered their estrous cycle, tending towards continuous estrus. The estrogen response from co-administration of genistein and methoxydidine appeared to be a cumulative effect of the individual actions of the two compounds. Methoxydidine (but not genistein) delayed prepuce separation (PPS) in male rats. When co-administered with methoxydidine, 800 ppm genistein enhanced the PPS-delaying effect of methoxydidine. Neither genistein nor methoxydidine treatment altered sex-specific locomotor patterns in male and female rats. To investigate the possible mechanisms of interaction between these two compounds during development, we conducted in vitro transcriptional activation assays based on estrogen receptor (ER) and androgen receptor (AR) responses using 2,2-bis-(p-hydroxyphenyl)-1,1,1-trichloroethane (HPTE), the major metabolite of genistein and methoxydidine. While in vitro results support estrogen-like effects of genistein and methoxydidine, as well as anti-androgen-like effects of methoxydidine, the reactivity of these compounds with ERα and ERβ did not predict the stronger estrogenic activity of methoxydidine in vivo compared to genistein; similarly, the in vitro reactions of HPTE and genistein with AR did not predict the enhancing effect of genistein on the PPS response of methoxydidine. The data from this study suggest that phytoestrogens can alter the toxicological behavior of other endogenous androgen compounds (EACs), and the interactions between these compounds can be highly complex and difficult to predict based on their in vitro steroid receptor reactivity.
...This study investigated the effects of soybean isoflavone genistein and the anti-estrogen tamoxifen (TAM) on the growth of estrogen (E)-dependent breast cancer (MCF-7) cells implanted in ovariectomized athymic mice. ...Six treatment groups were established: control group (C); 0.25 mg estradiol (E2) implantation group (E); E2 implantation + 2.5 mg TAM implantation group (2.5 TE); E2 implantation + 2.5 mg TAM implantation + 1000 ppm genistein group (2.5 TEG); E2 implantation + 5 mg tamoxifen implantation group (5 TE); and E2 implantation + 5 mg tamoxifen implantation group + 1000 ppm genistein group (5 TEG). Tamoxifen (2.5 TE and 5 TE) treatment inhibited E2-stimulated MCF-7 tumor growth in ovariectomized athymic mice. Dietary genistein can counteract/reverse the inhibitory effect of tamoxifen on MCF-7 tumor growth, reduce plasma E2 levels, and increase the expression of E-reactive genes (such as pS2, PR, and cyclin D1). …Postmenopausal women receiving tamoxifen treatment for E-reactive breast cancer should use dietary genistein with caution. The anticancer drug genistein can inhibit the growth of various malignant tumor cell lines. …The authors investigated the efficacy and possible mechanism of ionizing radiation (IR) combined with genistein treatment on cervical HeLa cells. The study found that the cell inhibition rate in the combined treatment group was significantly higher than that in the ionizing radiation (IR) or genistein treatment groups alone. After ionizing radiation (4 Gy) combined with genistein (40 μmol/L) treatment, the apoptosis index significantly increased, and the cell cycle arrested at the G2/M phase. Survivin mRNA expression increased after ionizing radiation (4 Gy), while it significantly decreased after the combined treatment. These results indicate that genistein enhances the radiosensitivity of cervical cancer HeLa cells, and its mechanism of action may include increasing apoptosis, reducing Survivin expression, and prolonging cell cycle arrest. Mice were treated with genistein (NPI031L; BIO-00; G2535; PTI G-4660; SIPI9764I) at a dose of 50 mg/kg/day for 28 consecutive days without significant weight loss or organ toxicity. Serum ALT, AST, creatinine, and BUN levels were all within the normal range [1]. The plasma protein binding rate of genistein in human plasma was approximately 85% as determined by balanced dialysis [4]. In long-term toxicity studies (30 days, 60 mg/kg/day, orally), rats did not show serious hematological or gastrointestinal toxicity, and no abnormal changes were found in the histopathological analysis of major organs [2].

[1]. J Biol Chem . 1987 Apr 25;262(12):5592-5.

[2]. Biochem Pharmacol . 1990 Jan 1;39(1):187-93.

[3]. J Biol Chem . 2003 Jan 10;278(2):962-7.

[4]. J Nutr . 2002 Mar;132(3):552S-558S.

[5]. Radiat Res . 2006 Jul;166(1 Pt 1):73-80.

Therapeutic Uses
Gentian root is considered beneficial for cardiovascular and bone health and can alleviate menopausal symptoms; however, studies on these endpoints are limited, inconsistent, or assess soy product intake rather than genistein alone. Interest in genistein has focused particularly on its role in menopausal treatment. This article reviews the major published studies to date on the efficacy of phytoestrogens in alleviating menopausal symptoms. Isoflavone-rich diets are associated with a reduced incidence of vasomotor attacks; the average genistein supplementation is approximately 50 mg/day. Studies have shown that total cholesterol and low-density lipoprotein cholesterol (LDL) levels decrease after phytoestrogen supplementation. Furthermore, bone mineral density (BMD) increases after 6 months of 90 mg isoflavone intake. Isoflavones may reduce the risk of breast cancer. The analyzed data confirm that soy extract supplementation, particularly genistein, has significant clinical efficacy in alleviating short-term menopausal symptoms and long-term effects, although the latter requires further confirmation.
/EXPTL/ ... /Authors/ This study evaluated and compared the effects of the phytoestrogen genistein, estrogen-progestin combination therapy (EPT), and placebo on hot flashes and endometrial thickness in postmenopausal women. ... Ninety healthy postmenopausal women aged 47 to 57 years were randomly assigned to receive one year of continuous EPT (n = 30; 1 mg 17β-estradiol combined with 0.5 mg norethindrone acetate), the phytoestrogen genistein (n = 30; 54 mg/day), or placebo (n = 30). Endometrial safety was assessed by transvaginal ultrasound at baseline, 6 months, and 12 months. ...Compared to placebo, after 3 months of genistein treatment, the mean daily hot flash score decreased significantly by 22% (95% CI: -38 to -6.2; P < 0.01); after 6 months, the mean daily hot flash score decreased significantly by 29% (95% CI: -45 to -13; P < 0.001); and after 12 months, the mean daily hot flash score decreased significantly by 24% (95% CI: -43 to -5; P < 0.01). Compared to placebo, after 3 months of estrogen treatment, the mean hot flash score decreased by 53% (95% CI: -79 to -26; P < 0.001); after 6 months, the mean decrease was 56% (95% CI: -83 to -28; P < 0.001); and after 12 months, the mean decrease was 54% (95% CI: -74 to -33; P < 0.001). No uterine side effects were observed in the subjects. This study confirms that genistein may have a positive effect on hot flashes without negatively impacting endometrial thickness, suggesting that this phytoestrogen may be a strategic treatment for postmenopausal symptoms. A growing body of in vitro and animal studies suggests that genistein may help prevent and treat certain cancers, primarily breast and prostate cancer. Currently, no clinical studies have been conducted to support or refute claims that genistein possesses anti-atherosclerotic properties or can be safely and effectively used as a natural estrogen replacement therapy. However, preliminary data suggest that soy isoflavones, including genistein, may help alleviate some menopausal-related issues such as osteoporosis and hot flashes. For more complete data on the therapeutic uses of genistein (6 types), please visit the HSDB record page. Drug Warnings: Genistein/genistein intake has been associated with hypothyroidism in some individuals. Women with estrogen receptor-positive tumors should use genistein/genistein supplements with caution and only under the guidance and supervision of a physician.
Prostate cancer patients should discuss the suitability of genistein/genistein supplements with their doctor before deciding to use them.
Pregnant and lactating women should avoid using genistein/genistein supplements until long-term safety studies are completed.
Use genistein with caution in postmenopausal women receiving tamoxifen treatment for estrogen-responsive breast cancer.

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Genistein (NPI031L; BIO-00; G2535; PTI G-4660; SIPI9764I) is a natural isoflavone found primarily in soybeans and other legumes. It has various pharmacological activities, including tyrosine kinase inhibition, topoisomerase II inhibition, and anti-angiogenic effects [1,2,3].
Because of its ability to inhibit tumor cell proliferation, induce apoptosis, and enhance the efficacy of radiotherapy, its potential in cancer prevention and treatment has been investigated [5].
In addition to its anticancer activity, genistein also has cardioprotective and antioxidant effects, but its low oral bioavailability limits its clinical application. Formulation optimization (e.g., nano-encapsulation) is being explored to improve its pharmacokinetic properties [4]

C15H10O5

270.24

270.052

C, 66.67; H, 3.73; O, 29.60

446-72-0

Genistein;446-72-0

5280961

Light yellow to yellow solid powder

1.5±0.1 g/cm3

555.5±50.0 °C at 760 mmHg

297-298 °C

217.1±23.6 °C

0.0±1.6 mmHg at 25°C

1.732

2.96

3

5

1

20

411

0

O1C([H])=C(C2C([H])=C([H])C(=C([H])C=2[H])O[H])C(C2=C(C([H])=C(C([H])=C12)O[H])O[H])=O

TZBJGXHYKVUXJN-UHFFFAOYSA-N

InChI=1S/C15H10O5/c16-9-3-1-8(2-4-9)11-7-20-13-6-10(17)5-12(18)14(13)15(11)19/h1-7,16-18H

5,7-dihydroxy-3-(4-hydroxyphenyl)chromen-4-one

NPI 031L; NPI031L; NPI-031L; BIO-300; G-2535; PTI-G-4660; SIPI-9764-I; PTIG-4660; SIPI-9764I; BIO300; G2535; PTIG4660; SIPI9764I; BIO 300; G 2535; PTI G 4660; SIPI 9764 I; PTIG 4660; SIPI 9764I; Genistein

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)

Solubility in Formulation 1: ≥ 3.75 mg/mL (13.88 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 37.5 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: ≥ 3 mg/mL (11.10 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 30.0 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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Solubility in Formulation 3: ≥ 3 mg/mL (11.10 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 30.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 5 mg/mL (18.50 mM) in 50% PEG300 50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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