Androgen receptor (AR) [1]

Lupeol is a useful AR mold for developing new therapeutics to cure human elk cancer (CaP). After 48 hours of treatment, lupeol (10-50 μM) inhibited the proliferation of androtrophic phenotype (ADPC) cells, namely LAPC4 and LNCaP cells, with IC50 values of 15.9 and 17.3 μM. Lupeol reduced the development of 22Rv_1 with an IC50 of 19.1 μM. Lupeol inhibits C4-2b cell growth at an IC50 of 25 μM. Lupeol has the ability to inhibit ADPC and CRPC. Phenotypic CaP cell growth capacity. Androgens, through AR activation, are known to promote CaP cell proliferation [1].

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Lupeol inhibited the growth of androgen-dependent prostate cancer (ADPC) cells LAPC4 (IC50=15.9 μM), LNCaP (IC50=17.3 μM), and castration-resistant prostate cancer (CRPC) cells C4-2b (IC50=25 μM) after 48 h treatment. It also inhibited 22Rv1 cells (IC50=19.1 μM). [1]
Lupeol (31-50 μM) significantly reduced R1881 (androgen analogue)-stimulated cell growth and proliferation (as measured by ³[H]thymidine uptake) in LAPC4 and LNCaP cells. [1]
Lupeol (added every 3rd day for 14 days) significantly reduced R1881-induced clonogenic potential of LNCaP and LAPC4 cells in soft agar. [1]
In AR-negative PC-3 cells, the growth suppression by Lupeol was not reversed by increasing R1881 concentration, while in AR-positive LNCaP cells, the suppression was partially reversed by increasing R1881 from 1.0 to 1.5 nM at 25 μM Lupeol but not at 50 μM. [1]
Lupeol (5-50 μM) significantly inhibited R1881-induced AR transcriptional activity (3.5-5 fold induction by R1881) in LAPC4 and LNCaP cells as assessed by luciferase-based ARE reporter assay. [1]
Lupeol decreased mRNA expression of PSA in a dose-dependent manner in LNCaP, LAPC4, 22Rv1 and C4-2b cells, and decreased protein levels of PSA in LNCaP, 22Rv1 and C4-2b cells. [1]
Lupeol decreased R1881-induced mRNA and protein levels of PSA in LAPC4 and LNCaP cells. [1]
Lupeol decreased secreted PSA levels in culture media from LNCaP and LAPC4 cells in a concentration-dependent manner. [1]
Lupeol (25-50 μM) competed with labeled R1881 for ligand binding domain of AR in LAPC4 and LNCaP cells. [1]
Lupeol treatment significantly reduced AR-DNA binding efficiency in LNCaP cells as assessed by EMSA. [1]
Lupeol decreased AR occupancy on ARE (androgen response elements) and ARB (AR binding sites) on AR-responsive genes (PSA, SGK-1, TIPARP, IL-6) in LNCaP cells and on PSA gene in C4-2b cells as assessed by ChIP assay. [1]
Lupeol reduced RNA polymerase II (RNA Pol II) occupancy at the PSA promoter in LNCaP cells (in presence of R1881) and in C4-2b cells (both in presence and absence of androgen). [1]
Lupeol sensitized C4-2b cells to bicalutamide: cells pre-treated with Lupeol (30-50 μM for 24 h) followed by bicalutamide (10 μM for 24 h) showed significant decrease in cell viability and PSA expression. [1]

One powerful medication that may reduce the tumorigenicity of CaP cells in the stroma is lupeol. Total circulatory PSA levels (via stromal tumor cell apoptosis) were assessed on day 56 of the research. At poststromal day 5 56, PSA levels in control rats with LNCaP and C4-2b tumors varied from 11.95 to 12.79 ng/mL, respectively. Conversely, rats treated with lopezol showed lower serum PSA levels, ranging from 4.25-7.09 ng/mL. Serum PSA levels were lower in tumor tissue from animals given lopeol than in control subjects [1].

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In athymic nude mice bearing LNCaP (ADPC) or C4-2b (CRPC) cell-derived tumors, intraperitoneal administration of Lupeol (40 mg/kg, 3 times per week) significantly inhibited tumor growth. At day 56 post-inoculation, average tumor volume in control mice was 1138 mm³ (LNCaP) and 1198 mm³ (C4-2b), while in Lupeol-treated mice it was 309 mm³ and 510 mm³, respectively. Approximately 60% (LNCaP) and 50% (C4-2b) of Lupeol-treated mice did not reach the endpoint (1000 mm³ tumor volume) by the end of week 12. [1]
Lupeol treatment reduced serum PSA levels in tumor-bearing mice: control animals had 11.95-12.79 ng/mL, while Lupeol-treated animals had 4.25-7.09 ng/mL. Immunoblot analysis of tumor tissues confirmed reduced PSA protein levels. [1]

In silico molecular docking: Lupeol was docked into the active region of AR (2PNU.pdb) using AutoDock4 and Sybyl programs. The binding energy was -10.66 Kcal/mol (~56 nM). Lupeol interacted with AR through 17 putative amino acid residues and 2 hydrogen bonds, surrounded by a hydrophobic pocket consisting of several hydrophobic side chains and 6 methionines (742,745,749,780,787,895) on 5 different helices, with hydrophilic interactions with Trp752 and Asn705. [1]
AR competitive ligand-binding assay: Cells were incubated with 1 nM ³[H]R1881 in the absence or presence of 0.1-100-fold molar excess of unlabeled competitor ligands (including Lupeol) for 90 min at 37°C. Bound ligands were extracted in ethanol for 30 min at room temperature and detected using a scintillation counter. Unlabeled R1881 and bicalutamide effectively competed ³[H]R1881 binding; Lupeol at 25-50 μM also competed for the ligand binding domain of AR. [1]
Electrophoretic mobility shift assay (EMSA): Nuclear lysates from cells treated with Lupeol were incubated with biotin-labeled AR oligonucleotide (sense: 5'-TGC AGA ACA GCA AGT GCT AGC-3'; anti-sense: 5'-GCT AGC ACT TGC TGT TCT GCA-3'), and DNA binding was assessed. Controls included biotin-EBNA control DNA, EBNA extract, and competition with excess unlabeled EBNA DNA. [1]
Chromatin immunoprecipitation (ChIP) assay: Cells were treated with Lupeol, then cross-linked, lysed, and chromatin was immunoprecipitated with anti-AR antibody or anti-RNA Pol II antibody. AR occupancy on ARE/ARB sites of PSA, SGK-1, TIPARP, IL-6 genes, and RNA Pol II occupancy at PSA promoter were measured by PCR. [1]

Cell viability assay (MTT): Cells (60-70% confluent) were treated with Lupeol (10-50 μM) for 48 h in complete growth medium. MTT solution was added, incubated, and formazan crystals dissolved in DMSO, measured at 570 nm. For combination studies, cells were treated with R1881 (1 nM), bicalutamide (10 μM), or R1881+Lupeol for 48 h. For sensitization, C4-2b cells were treated with Lupeol for 24 h then bicalutamide (10 μM) for another 24 h before MTT. [1]
³[H]thymidine incorporation assay: Cells were treated as indicated, then pulsed with ³[H]thymidine, and incorporated radioactivity was measured. [1]
Colony formation assay: Cells were seeded in soft agar, treated with R1881 (1 nM daily) and/or Lupeol (added every 3rd day for 14 days), then colonies were counted. [1]
AR transcriptional activity reporter assay: LAPC4 and LNCaP cells were transfected with ARE-Luc plasmid (200 ng/well) for 24 h, then treated with R1881 (1 nM), bicalutamide (10 μM), Lupeol (10-50 μM), or combinations for 48 h. Luciferase activity was measured using dual luciferase assay kit. [1]
Real-time PCR for PSA expression: Cells were treated with Lupeol, total RNA extracted, and real-time PCR performed using PSA primers (sense: 5'-GTGCTTGTCGCTCTCGCT-3'; anti-sense: 5'-CAGCAAGATCACGTCTTTG-3'). [1]
Immunoblot analysis: Cell lysates or tumor tissue lysates were subjected to SDS-PAGE, transferred to membranes, and probed with anti-PSA and anti-AR antibodies. β-actin was used as loading control. [1]
PSA secretion assay: Culture media from treated LNCaP and LAPC4 cells were collected and secreted PSA levels were measured. [1]

Athymic nude mice bearing C4-2b and LNCaP cell-originated tumors were treated intraperitoneally with Lupeol (40 mg/kg; 3 times per week) for up to 12 weeks. Tumor growth was measured as volume over time. Mice were monitored for body weight, food intake, and signs of toxicity. At day 56 post-inoculation (when 100% of control animals reached tumor volume >1000 mm³), tumors and serum were collected for analysis. Corn oil was used as vehicle control. [1]
For bioavailability studies, mice received a single intraperitoneal administration of Lupeol (200 mg/kg), and serum was collected at 4 h and 8 h post-administration. Alternatively, mice bearing tumors were treated with Lupeol (40 mg/kg) for up to 8 weeks, and serum was collected. [1]

Lupeol was detected in serum by HPLC with a retention time of 13.4 min, matching the standard, indicating stability and bioavailability. [1]
After a single intraperitoneal administration of Lupeol (200 mg/kg), serum levels were 3.08 μM at 4 h and 5.22 μM at 8 h post-administration. [1]
In mice bearing tumors and receiving repeated Lupeol treatment (40 mg/kg, 3 times/week for 4-8 weeks), serum Lupeol levels reached 10-20 μM. [1]

Interactions
In in vivo studies using an orthotopic metastatic nude mouse oral and tongue squamous cell carcinoma model, 2 mg/mouse lupeol significantly reduced tumor volume and inhibited local metastasis, showing superior efficacy compared to cisplatin alone. Lupeol, when used in combination with low-dose cisplatin, exhibited significant synergistic cytotoxicity without side effects…
…This study…used an alkaline melting method to investigate the effect of lupeol on 7,12-dimethylbenzo[a]anthracene (DMBA)-induced DNA strand breaks in mouse skin. Samples were taken at 24, 48, 72, and 96 hours before or after a single topical application of DMBA (100 μg/mouse), and incremental doses of lupeol (50-200 μg/mouse) were administered topically. Both pretreatment and posttreatment with lupeol showed significant preventative effects against DMBA-induced DNA strand breaks (p<0.001), in a dose- and time-dependent manner. Pretreatment with 200 μg/mouse prevented 56.05% of DNA strand breaks after 96 hours; posttreatment prevented 43.26% of DNA strand breaks… This study evaluated the antigenotoxic effects of the triterpenoid compounds lupeol and mango pulp extract (MPE) in Swiss albino mice. The known mutagen benzo[a]pyrene (B[a]P) was administered intraperitoneally at a single dose of 100 mg/kg body weight. Seven days prior to B[a]P administration, mice were pretreated with lupeol (1 mg/mouse) and MPE (1 mL, 20%) via gavage. All animals were sacrificed at 24-hour sampling time, and chromosomal damage and micronucleus induction in their bone marrow tissue were analyzed. In the benzo[a]pyrene (B[a]P) treatment group, a significant increase in chromosomal aberrations and micronuclei, along with a decreased mitotic index, was observed. In the groups supplemented with lupeol or MPE, B[a]P-induced chromosome breakage was significantly reduced. Compared with the B[a]P-treated group, both lupeol and MPE reduced the incidence of abnormal cells and micronuclei. Furthermore, the anti-cytotoxic effects of lupeol or MPE were also significant, manifested by a significant increase in the mitotic index. Therefore, the results of this study indicate that lupeol and mango pulp extract (MPE) have a protective effect against benzo[a]pyrene (B[a]P)-induced chromosome breakage in Swiss albino mice. This study aimed to investigate the antioxidant effect of lupeol/mango pulp extract (MPE) on testosterone-induced oxidative stress in the prostate of male Swiss albino mice. …For 15 consecutive days, mice were orally administered lupeol (1 mg/mouse) and MPE (1 mL [20% w/v]/mouse), respectively, while simultaneously receiving a subcutaneous injection of testosterone (5 mg/kg body weight). At the end of the study, the prostate was dissected, and reactive oxygen species (ROS) levels, lipid peroxidation, and the activities of antioxidant enzymes (catalase, superoxide dismutase, glutathione reductase, and glutathione S-transferase) were measured. Results: In testosterone-treated animals, increased ROS led to depletion of antioxidant enzymes and increased lipid peroxidation in the mouse prostate. However, lupeol/MPE treatment resulted in decreased ROS levels, and lipid peroxidation and antioxidant enzyme levels returned to normal. …These results indicate that lupeol/MPE effectively combats oxidative stress-induced prostate cell damage in mice…
For more complete interaction data (14 items in total) on lupeol, please visit the HSDB record page.

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Non-human toxicity values
Rats oral LD50 >2 g/kg /see table/
Mice oral LD50 >2 g/kg /see table/

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Lupeol treatment (40 mg/kg, 3 times/week intraperitoneally) did not cause any loss in body weight, change in food intake, or exhibit apparent signs of toxicity in athymic nude mice. [1]

[1]. Lupeol, a novel androgen receptor inhibitor: implications in prostate cancer therapy. Clin Cancer Res. 2011 Aug 15;17(16):5379-91.

Lupeol is a pentacyclic triterpenoid compound, a derivative of lupene, in which the hydrogen atom at the 3β position is replaced by a hydroxyl group. It is found in the seed coat of lupin seeds, as well as in the latex of fig and rubber trees. Additionally, it is present in many edible fruits and vegetables. It has anti-inflammatory properties and is a plant metabolite. Lupeol is a secondary alcohol belonging to the pentacyclic triterpenoid class. It is derived from the hydride of lupene. Lupeol has been studied for its use in treating acne. It has been reported to be found in tea (Camellia sinensis), holly leaf erythrorhiz (Acanthus ilicifolius), and other organisms with relevant data. See also: Calendula officinalis flower (part).

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Mechanism of Action
Lupinol is a triterpenoid compound found in fruits and vegetables. It selectively induces the death of large numbers of head and neck squamous cell carcinoma (HNSCC) cells, but has little effect on normal tongue fibroblast cell lines cultured in vitro. Downregulation of NF-κB is considered the main mechanism by which lupinol combats HNSCC. Lupinol not only inhibits tumor growth but also suppresses HNSCC cell invasion by reversing NF-κB-dependent epithelial-mesenchymal transition. Lupinol has a synergistic effect with cisplatin, enabling HNSCC cell lines with high NF-κB activity in vitro to become chemosensitized by cisplatin.

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Lupeol (Lup-20(29)-en-3b-ol) is a triterpene found in fruits, vegetables, and medicinal plants, possessing strong antioxidant, anti-inflammatory, anti-arthritic, anti-mutagenic and anti-malarial activities. [1]
Mechanism: Lupeol acts as a competitive antagonist for AR, blocking AR binding to androgen, inhibiting AR-DNA binding, reducing AR occupancy on ARE/ARB sites of AR-responsive genes (PSA, TIPARP, SGK, IL-6), and inhibiting RNA Pol II recruitment to target genes. It also sensitizes CRPC cells to anti-hormone therapy (bicalutamide). [1]
Lupeol is a diet-derived agent, non-toxic, and bioavailable in vivo, suggesting translational potential for treating both androgen-dependent and castration-resistant prostate cancer. [1]

C30H50O

426.729

426.386

545-47-1

259846

White to off-white solid powder

1.0±0.1 g/cm3

488.1±14.0 °C at 760 mmHg

215-216ºC

216.9±12.4 °C

0.0±2.8 mmHg at 25°C

1.516

10.98

1

1

1

31

766

10

CC(=C)[C@@H]1CC[C@]2([C@H]1[C@H]3CC[C@@H]4[C@]5(CC[C@@H](C([C@@H]5CC[C@]4([C@@]3(CC2)C)C)(C)C)O)C)C

MQYXUWHLBZFQQO-QGTGJCAVSA-N

InChI=1S/C30H50O/c1-19(2)20-11-14-27(5)17-18-29(7)21(25(20)27)9-10-23-28(6)15-13-24(31)26(3,4)22(28)12-16-30(23,29)8/h20-25,31H,1,9-18H2,2-8H3/t20-,21+,22-,23+,24-,25+,27+,28-,29+,30+/m0/s1

Lup-20(29)-en-3-ol, (3-beta)-

NSC 90487Monogynol BLupeol β-ViscolClerodol Fagarasterol Lupenol

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)

Ethanol : ~14.29 mg/mL (~33.49 mM)
DMSO : ~2 mg/mL (~4.69 mM)

Solubility in Formulation 1: ≥ 1.43 mg/mL (3.35 mM) (saturation unknown) in 10% EtOH + 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 14.3 mg/mL clear EtOH + stock solution to 900 μL of corn oil and mix well.

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Solubility in Formulation 2: 20 mg/mL (46.87 mM) in Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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