Product name: Cyasterone
Synonym name:
Catalogue No.: BP0432
Cas No.: 17086-76-9
Formula: C₂₉H₄₄O₈
Mol Weight: 520.663
Botanical Source: Achyranthes bidentata Bl.
Physical Description:
Type of Compound: Steroids

Purity: 95%~99%
Analysis Method: HPLC-DAD or/and HPLC-ELSD
Identification Method: Mass, NMR
Packing: Brown vial or HDPE plastic bottle

Storage: Store in a well closed container, protected from air and light. Put into refrigerate or freeze for long term storage.
Whenever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20℃. Generally, these will be useable for up to two weeks.

The product could be supplied from milligrams to grams
Inquire for bulk scale.

Description:

Cyasterone, a natural EGFR inhibitor, can inhibit growth of A549 and MGC823 cells, via regulating EGFR signaling pathway, it maybe a promising anti-cancer agent; it also has antifeeding activity.

References:

Insect Sci., 2001, 8(3):233-9.    

Effects of cyasterone on growth and development of diamondback moth, Plutella xylostella (L.)   

Cyasterone is the main phytoecdysteroid component from cultured plants of Ajuga nipponensis, and common in other Ajuga species. 

METHODS AND RESULTS:

Results showed that when treated with 50 mg/L Cyasterone, the egg hatching of diamondback moth was retarded. The total percentage of eggs hatched in treatment was 94.7%, significantly less than that in control, which was 100%. Low toxicity and good antifeeding activity to the 3rd instar larvae were exhibited, which are dependent upon concentration. The growth and development of diamondback moth were affected by Cyasterone. It was found that, at concentrations higher than 50 mg/L Cyasterone, the larval growth and egg production were inhibited; while promoted at lower concentrations. 

CONCLUSIONS:

This dual role of Cyasterone might be attributed to its hormonal activity and antifeeding activity, which resulted in poor nutrition. The pupation and eclosion were hindered by treatment of larvae. This insect species was more susceptible to Cyasterone than 20-hydroxyecdysone.    

Biomed. Pharmacother., 2016, 84:330-9.    

Anti-proliferation effects, efficacy of cyasterone in vitro and in vivo and its mechanism.  

Cyasterone was demonstrated potential inhibition effect in mouse skin carcinoma cells in published report. However, the molecular mechanisms of the Cyasterone on cells remain unknown. 

METHODS AND RESULTS:

Herein, we investigated the effects of Cyasterone-induced apoptosis in A549 and MGC823 cells in vitro. MTT assay showed that Cyasterone caused a significantly decreasing of the proliferation of A549 and MGC823 cells in a time-and dose-dependent manner with IC50 values of 38.50±3.73μg/mL on A549 cells and 32.96±1.24μg/mL on MGC823 cells at 48h, respectively. Hoechst staining and TUNEL staining results indicated the quintessential apoptosis features in immunofluorescence image. Apoptosis and cell cycle were determined by flow cytometry. Cyasterone treatment triggered inhibition of epidermal growth factor receptor- phosphatidylinositol 3 kinase/protein kinase B (EGFR-AKT) signaling pathways and activation of P38 pathways. Furthermore, Cyasteroneinhibited MGC823 cells xenografted tumor growth in vivo with few changes in body weights. 

CONCLUSIONS:

In conclusion, our findings provide the evidence that Cyasterone inhibits growth of A549 and MGC823 cells, via regulating EGFR signaling pathway. Our results indicated that Cyasterone, a natural EGFR inhibitor, maybe a promising anti-cancer agent.    

Storage conditions：Short-term storage at 2~8℃, long-term storage at -20 ~ -80℃

Introduction/Overview

Malignant tumor, especially breast cancer, is a serious disease threatening human health worldwide. Despite continuous advancements in strategies such as surgery, radiotherapy, chemotherapy, and targeted therapy, drug resistance, recurrence, and metastasis of tumors remain severe challenges in clinical practice. Therefore, searching for efficient and low toxicity novel anti-tumor lead compounds from natural products has always been an important direction in drug development. Cyasterone (CAS No. 17086-76-9), as a natural steroid compound isolated from traditional medicinal plants, has attracted extensive attention of pharmacology researchers in recent years due to its remarkable anti-tumor activity, especially its inhibitory effect on breast cancer. Its unique chemical structure enables it to act on multiple key targets such as epidermal growth factor receptor (EGFR), exerting anti proliferative effects through multiple mechanisms such as inducing cell apoptosis and cell cycle arrest. This article aims to provide a systematic review of the chemical properties, plant sources, pharmacological activities, molecular mechanisms of action, pharmacological evaluation, and clinical application potential of naringenin, in order to provide comprehensive scientific references for the in-depth research and development of this natural product.

Chemical structure and physicochemical properties

Cup amaranthrone belongs to the class of ecdysterone compounds, and its chemical name is (2 β, 3 β, 5 β, 22R) -2,3,14,20,22,25-hexahydroxycholestan-7-en-6-one. The molecular formula is C29H44O8 and the molecular weight is 520.6630. Its core structure is the steroid nucleus, with a ketone group at position C-6 and a double bond at position C-7. Multiple hydroxyl groups are connected at positions C-2, C-3, C-14, C-20, C-22, and C-25, with the configuration at position C-22 being R-type. The structural characteristics of this polyhydroxylation have a decisive impact on its biological activity and physicochemical properties.

From the analysis of parameters related to drug properties, the lipid water partition coefficient (LogP) of amaranth ketone is 1.6355, indicating that it has a certain lipophilicity, but not highly hydrophobic, which is beneficial for its penetration into cell membranes. Its topological polar surface area (TPSA) is as high as 144.52 Å ², mainly attributed to the numerous hydroxyl and ketone groups in the molecule, which enable it to form abundant hydrogen bonds. Correspondingly, its water solubility value is 0.0852, belonging to the category of slight solubility, which suggests that solubilization strategies may need to be considered in formulation development. Preliminary pharmacokinetic predictions indicate that the ability of cup-shaped ketone to penetrate the blood-brain barrier is relatively low, which to some extent limits its direct effects on central nervous system tumors, but may also reduce potential central nervous system side effects. Importantly, its hERG inhibition risk is' no ', and the Ames test result is 0.0, indicating a low risk of cardiac toxicity and genetic toxicity, providing preliminary positive signals for its safety as a lead compound.

Plant sources and extraction methods

Cup amaranth ketone is mainly isolated from plants of the Labiatae family, including the genus Caryophyllum Creeping Muscle Bone Grass（Ajuga decumbens Thunb.）， This plant is commonly used as a traditional herb in East Asia, especially in China, Japan, and South Korea, to treat inflammation, fever, and liver disease. In addition, in plants of the same genus such as Ajuga decumbens Cupressterone has also been found in certain plants of Ajuga bracteosa and other families and genera, but creeping muscle grass remains its main and most commonly studied natural source.

Organic solvent extraction is commonly used to extract chalcone from plant materials. The classic process includes crushing the dried creeping muscle grass whole plant, first degreasing it with petroleum ether or n-hexane to remove weak polar impurities such as chlorophyll and oil. Subsequently, medium polarity solvents such as methanol, ethanol, or ethyl acetate are used for repeated leaching or reflux extraction. After vacuum concentration, the obtained crude extract was separated and purified using various chromatographic techniques, such as silica gel column chromatography, reverse phase C18 column chromatography, high performance liquid chromatography (HPLC), and preparative thin layer chromatography (PTLC). By using nuclear magnetic resonance (NMR), mass spectrometry (MS), and comparison with standard samples, high-purity cup-shaped ketone monomers can be ultimately identified and obtained. With the development of green extraction technology, methods such as ultrasound assisted extraction and microwave-assisted extraction have also been attempted to improve extraction efficiency and reduce solvent consumption.

Pharmacological activity research

The most notable pharmacological activity of amaranth ketone is its wide range of antitumor activity Studies have shown that it has significant concentration dependent growth inhibitory activity on a variety of human tumor cell lines, especially breast cancer cells (such as MCF-7, MDA-MB-231, T47D, etc.). Its anti-tumor effect is mainly achieved through the following two core ways:

1. Inducing cell apoptosis Cuplane ketone treatment can significantly upregulate the expression of pro apoptotic proteins (such as Bax) and downregulate the level of anti apoptotic proteins (such as Bcl-2), leading to a decrease in mitochondrial membrane potential, release of cytochrome C, and activation of the Caspase cascade reaction, ultimately triggering typical cell apoptosis. Morphologically, cell shrinkage, chromatin condensation, and the formation of apoptotic bodies can be observed.
2. Inducing cell cycle arrest Cuplane ketone can block tumor cells at specific phases of the cell cycle, most commonly in the G1 or G2/M phase. This blockade is closely related to the inhibition of the expression of key cyclins (such as Cyclin D1, Cyclin B1) and cyclin dependent kinases (CDKs), as well as the upregulation of cyclin dependent kinase inhibitors (such as p21). The stagnation of the cell cycle process prevents the unlimited proliferation of tumor cells.

In addition to its direct cytotoxic effects, cup-shaped ketone has also been reported to have anti-inflammatory and antioxidant Activity, these characteristics may indirectly contribute to its anti-tumor effect, as chronic inflammation and oxidative stress are important promoting factors for tumor occurrence and development.

Mechanism of action and molecular targets

The antitumor effect of caliamarone involves a complex regulatory network of multiple targets and pathways, especially in breast cancer. Its main molecular targets and mechanisms of action include:

1. EGFR signaling pathway inhibition Cuplane ketone has been identified as a natural EGFR inhibitor. It can bind to the kinase domain of EGFR, inhibit its autophosphorylation and downstream signaling pathways such as RAS/RAF/MEK/ERK and PI3K/Akt/mTOR, thereby suppressing tumor cell proliferation, survival, and migration.
2. AMPK (PRKAA1) activation Adenosine activated protein kinase (AMPK) is a core regulatory factor in cellular energy metabolism. Cup amaranth ketone can activate AMPK, thereby inhibiting its downstream synthetic metabolic pathways (such as mTORC1), inducing autophagy, and regulating cell growth and metabolic reprogramming, which is crucial for inhibiting tumor growth.
3. STAT3 signal suppression Signal transduction and transcription activator 3 (STAT3) is an important oncogenic transcription factor. Cuplane ketone can inhibit the phosphorylation (Tyr705 site) and nuclear translocation of STAT3, thereby downregulating the expression of its target genes (such as Bcl-2, Cyclin D1, MMP-2), promoting apoptosis, and inhibiting proliferation and invasion.
4. Regulation of estrogen receptor beta (ESR2)For estrogen receptor positive breast cancer, amaranthine may interfere with estrogen dependent growth signals by regulating the expression or activity of ESR2.
5. Affects apoptosis related proteins Directly regulating the balance of Bcl-2 family proteins (inhibiting BCL2 and promoting pro apoptotic proteins) is the core link in the mitochondrial pathway apoptosis induced by kaempferol.
6. Inhibition of invasion and metastasis related factors Cup amaranth ketone can downregulate the expression and activity of matrix metalloproteinase-2 (MMP-2), thereby inhibiting the degradation and invasion ability of tumor cells to extracellular matrix. The potential impact on microtubule associated protein tau (MAPT) may also involve cytoskeleton remodeling.
7. Reverse multidrug resistance (MDR) potential: Studies have shown that amaranth sterone may inhibit the function of ATP binding box transporter superfamily members such as P-glycoprotein (ABCB1) and breast cancer resistant protein (ABCG2), reduce the excretion of chemotherapy drugs from the cell to the outside, and thus enhance the sensitivity of drug resistant tumor cells to conventional chemotherapy drugs.
8. Other targets Inhibition of protein kinase C alpha (PRKCA) may affect various cellular signals; The inhibitory activity of tyrosinase (TYR) suggests its potential application value in other tumors such as melanoma.

In summary, by synergistically acting on multiple key targets mentioned above, cup-shaped ketone forms a comprehensive network that inhibits tumor growth, promotes apoptosis, blocks cycles, inhibits invasion, and may reverse drug resistance.

Evaluation of drug properties and pharmacokinetics

Despite exhibiting good anti-tumor activity in vitro and some in vivo models, the pharmacological potential of cupped amaranth ketone as a drug candidate still needs to be comprehensively evaluated.

Pharmacokinetic aspects Currently, there is relatively limited in vivo pharmacokinetic research data available for publicly available systems. Based on its physicochemical properties (moderate LogP value, high TPSA, low water solubility), it can be inferred that its oral bioavailability may face challenges: it may be limited in gastrointestinal absorption and susceptible to first pass effects. Its blood-brain barrier permeability is low, as mentioned earlier. The key parameters of the metabolic pathway, main metabolites, half-life, distribution volume, and excretion mode of sitosterol in vivo need to be elucidated through more in-depth animal experiments (such as pharmacokinetic studies in rats and mice). These studies are crucial for determining appropriate routes of administration (such as oral administration, injection) and dosage form design.

safety evaluation The preliminary computer predictions (hERG inhibition negative, Ames test negative) provide a good starting point. But comprehensive preclinical toxicology studies must be conducted, including acute toxicity, subchronic toxicity, genetic toxicity, reproductive toxicity, etc., to clarify its safe dose window.

Pharmaceutical Challenge Its micro solubility is the main obstacle to formulation development. Advanced drug delivery technologies are needed to improve its solubility and bioavailability, such as making nanocrystals, liposomes, polymer micelles, cyclodextrin inclusion complexes, or solid dispersions.

Clinical application prospects and prospects

As a multi-target anti-tumor natural product, cupped amaranth ketone has broad clinical application prospects, but also faces many challenges.

Potential application directions：
1. Adjuvant or alternative drugs for breast cancer treatment In view of its effectiveness on a variety of breast cancer cell lines, especially for the regulation of EGFR signaling pathway, STAT3 pathway and AMPK pathway, caliamarone is expected to be developed as a potential drug for the treatment of triple negative breast cancer (lack of clear targets) or breast cancer resistant to existing targeted drugs.
2. Chemosensitizer It reverses the multidrug resistance potential mediated by ABCB1/ABCG2, making it possible to use it in combination with traditional chemotherapy drugs such as doxorubicin and paclitaxel to improve chemotherapy efficacy, reduce required doses and side effects.
3. Combination therapy strategy Combined use with existing targeted drugs (such as EGFR inhibitors, CDK4/6 inhibitors) or immune checkpoint inhibitors may produce synergistic effects and overcome the limitations of monotherapy.
4. Other types of cancer The mechanism of action suggests that cupped amaranth steroids may also have therapeutic potential for other solid tumors that overly rely on the same signaling pathway, such as lung cancer, liver cancer, and colon cancer.

Future research prospects：
1. In depth mechanism research It is necessary to use techniques such as gene knockout/knockdown, chromatin immunoprecipitation (ChIP), proteomics, etc. to more accurately elucidate the direct interaction details and downstream networks between quercetin and various targets.
2. Systematic pharmacokinetics and toxicology research Conduct standardized pharmacokinetic and toxicological studies in animals to obtain key data on absorption, distribution, metabolism, excretion (ADME), and safety.
3. Structural optimization and derivative development Based on the parent nucleus structure of amaranthrone, reasonable chemical modifications are carried out to improve its activity, solubility, metabolic stability, and targeting, and reduce potential toxicity.
4. Advanced delivery system research and development Actively developing a new nano drug delivery system suitable for cupped amaranth steroid ketone to solve its drug development bottleneck.
5. High quality preclinical and clinical research Ultimately, rigorous preclinical efficacy validation (such as human tumor xenograft PDX models) and gradual clinical trials need to be designed to evaluate their true efficacy and safety in humans.

Conclusion

Cup amaranth steroid ketone is a natural molting steroid ketone with important research value discovered from the traditional medicinal plant creeping muscle bone grass. It effectively induces tumor cell apoptosis and cycle arrest by inhibiting EGFR, activating AMPK, inhibiting STAT3, regulating Bcl-2 family proteins and other multi-target mechanisms, and demonstrates the potential to reverse multidrug resistance. Although there are challenges in drug formulation, especially in solubility and systemic pharmacokinetics, the preliminary safety prediction results are positive. In the future, through in-depth molecular mechanism analysis, systematic preclinical efficacy and safety evaluation, reasonable structure optimization and innovative drug delivery technology development, caliamarone is expected to be successfully transformed into a new candidate drug or adjuvant for the treatment of malignant tumors such as breast cancer, injecting new vitality into the field of natural product anti-tumor drug research and development.