Product name: ent-11β-Hydroxyatis-16-ene-3,14-dione
Synonym name:
Catalogue No.: SBP03220
Cas No.: 1092103-22-4
Formula: C₂₀H₂₈O₃
Mol Weight: 316.441
Botanical Source:
Physical Description:
Type of Compound: Diterpenoids

Purity: 95%~99%
Analysis Method: HPLC-DAD or/and HPLC-ELSD
Identification Method: Mass, NMR
Packing: Brown vial or HDPE plastic bottle
The product could be supplied from milligrams to grams. Inquire for bulk scale.
We provide solution to improve the water-solubility of compounds, thereby facilitating the variety of activity tests and clinic uses.

For Reference Standard and R&D, Not for Human Use Directly.

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

Introduction/Overview

Natural products, as an important source of drug discovery, have played an indispensable role in the long history of human fight against diseases. From the classic analgesic morphine to the antimalarial artemisinin, the chemical diversity found in nature provides endless inspiration for modern drug development. Diterpenes, as a class of structurally diverse and biologically active natural products, have always been a hot topic in medicinal chemistry and pharmacology research. Among them, it originates from the Aconitum genus（Aconitum）Atisone type diterpenes from various plants have attracted much attention due to their unique [3.2.1] bicyclic skeleton and significant biological activities, especially anti-inflammatory, analgesic, and anti-tumor activities.

Ent-11 β - Hydroxyatis-16-ene-3,14-dione (hereinafter referred to as "the compound"), CAS number 1092103-22-4, is a typical diterpenoid compound of the alteson type. Its chemical structure is characterized by a highly oxidized tetracyclic skeleton, containing an alpha, beta unsaturated ketone structural unit (C-16 ene conjugated with C-14 ketone), as well as a C-3 carbonyl group and a C-11 beta hydroxyl group. This unique combination of functional groups endows the compound with complex chemical properties and potential biological activity. In recent years, research on this compound has gradually deepened, especially in the field of anti-inflammatory. Its potential to regulate multiple key inflammatory signaling pathways (such as NF - κ B, STAT3) and affect multiple inflammatory mediators (such as IL-6, TNF - α) has been preliminarily revealed.

This review aims to systematically review and summarize the research progress on ent-11 β - Hydroxyatis-16-ene-3,14-dione. The article will first elaborate on its chemical structure and physicochemical properties, followed by an introduction to its plant origin and extraction methods, with a focus on its pharmacological activities such as anti-inflammatory and analgesic effects, and further explore its mechanism of action and molecular targets. On this basis, combined with its pharmacological parameters, the pharmacokinetic characteristics, clinical application prospects, and challenges of this natural product are discussed, in order to provide comprehensive scientific basis for its further development and utilization.

Chemical structure and physicochemical properties

chemical structure

Ent-11 β - Hydroxyatis-16-ene-3,14-dione belongs to the atisane type of diterpenes, and its core skeleton consists of four rings: one cyclohexane (A ring), one cyclohexane (B ring), one cyclopentane (C ring), and one cyclobutane (D ring), forming a unique [3.2.1] bridged ring system. Its systematic naming clearly reveals its structural features:
- ent-The compound is a natural enantiomer, and its absolute configuration is opposite to that of the synthetic standard.
- 11β-Hydroxy There is a β - oriented hydroxyl group (- OH) at position C-11.
- atis-16-ene This indicates that its parent nucleus is atezane and there is a double bond (ene bond) between positions C-16 and C-17.
- 3,14-dione There is a ketone group (C=O) at positions C-3 and C-14 respectively.

The molecular formula of this compound is C ₂₀ H ₂₈ O3, with a molecular weight of 316.4410 g/mol. The key functional groups in its structure include:
1. α. β - Unsaturated Ketone System The C-16=C-17 double bond is conjugated with the C-14 ketone group to form an α, β - unsaturated ketone structural unit. This structure is a key pharmacophore in many biologically active natural products, often associated with Michael addition reaction activity, and may participate in covalent modification of target proteins.
2. C-3 keto group The carbonyl group located on the A ring may affect the polarity and hydrogen bonding ability of the molecule.
3. C-11 β - hydroxy group The presence of this hydroxyl group increases the water solubility of the molecule and may act as a hydrogen bond donor or acceptor, participating in interactions with target proteins.

Physicochemical properties

According to calculations and experimental data, the compound exhibits the following key physicochemical properties:
- molecular weight：316.4410 Da， Belonging to the category of small molecule compounds, it is beneficial for cell membrane permeation and oral absorption.
- Lipid water partition coefficient (LogP): 2.4144. This value indicates that the compound has moderate lipid solubility, which can dissolve in lipid environments (such as cell membranes) and has a certain degree of water solubility, which is beneficial for distribution and transport in the body. A LogP value between 2-3 is generally considered the ideal range for oral medication.
- Topological Polarity Surface Area (TPSA): 54.37 Å ². TPSA is an important parameter for predicting the ability of molecules to passively penetrate cell membranes. Generally, molecules with TPSA less than 140 Å ² have good oral bioavailability, while molecules with TPSA less than 60-70 Å ² are more likely to cross the blood-brain barrier. The TPSA of this compound is 54.37 Å ², indicating its high oral absorption potential and potential central nervous system (CNS) activity.
- Water solubility：0.0586 mg/mL。 The low value indicates that the compound has limited solubility in water and belongs to poorly soluble drugs. This may be a potential factor limiting its bioavailability after oral administration.
- Blood-brain barrier (BBB) permeability Predicted as' high '. Combined with its moderate LogP and lower TPSA, this compound is likely to penetrate the blood-brain barrier, providing possibilities for its therapeutic application in central nervous system related diseases such as neuroinflammation and pain.
- HERG inhibition Predicted as' no '. The hERG (human Ether - à - go Related Gene) potassium ion channel is an important target for assessing cardiac toxicity. This prediction reduces the likelihood of triggering arrhythmia risks such as prolonged QT interval in the heart, and is a positive pharmacological indicator.
- Ames test The predicted result is 0.0 (negative). The Ames test is used to detect the mutagenicity of compounds. The negative result indicates that the compound did not show significant genetic toxicity in the bacterial recovery mutation test, and the preliminary safety is good.

Plant sources and extraction methods

Plant-based

Ent-11 β - Hydroxyatis-16-ene-3,14-dione is mainly from the Aconitum genus in the Ranunculaceae family（Aconitum）Separated from plants. Aconitum plants are an important component of traditional Chinese medicine, such as Chuanwu（Aconitum carmichaelii）Caowu（Aconitum kusnezoffii）It is commonly used to treat inflammatory diseases such as rheumatism, rheumatism, and traumatic injuries. This compound was originally derived from Aconitum Found in the rhizomes of plants. In addition, there are also reports from other closely related plants, such as the genus Quercus（Delphinium）Separated from plants. These plants usually grow in high-altitude areas and have complex chemical compositions, rich in various diterpenoid alkaloids and demethylated diterpenoid alkaloids, which belong to the relatively low content but unique structure of non alkaloid diterpenoid components.

Extraction and Separation Methods

Due to the low content of this compound in plants and its frequent coexistence with a large number of structurally similar diterpenes, a systematic method is required for its extraction, separation, and purification. The typical process is as follows:

1. Raw material preparation and extraction：

   • raw material Usually, dried and crushed roots and stems of Aconitum plants are used.
   • extraction solvent According to the moderate polarity of the compound (LogP ≈ 2.4), solvents with low to high polarity are often used for gradient extraction. The most commonly used method is to use 95% ethanol or methanol for cold soaking or reflux extraction to obtain the total extract. Sometimes ethyl acetate or chloroform is used for targeted extraction to improve the enrichment efficiency of target compounds.
   • extraction method Cold soaking (soaking at room temperature for several days) or heating reflux extraction (several hours) are commonly used methods. Ultrasonic assisted extraction or microwave-assisted extraction can improve efficiency and shorten time.

2. Preliminary separation and enrichment：

   • solvent extraction Suspend the total extract in water and perform liquid-liquid extraction using solvents of different polarities such as petroleum ether, ethyl acetate, n-butanol, etc. This compound is usually enriched in the ethyl acetate extraction layer or chloroform extraction layer due to its equipolarity.
   • Acid-base treatment Due to the high content of alkaloids in Aconitum plants, acid-base treatment methods (such as extracting with dilute acid, alkalizing, and then extracting with organic solvents) are often used to remove alkaloids and enrich non alkaloid components (such as the compound).

3. Chromatographic Separation and Purification：

   • Positive phase silica gel column chromatography This is the most essential means of separation. Usually, different ratios of petroleum ether ethyl acetate or chloroform methanol mixed solvents are used as the mobile phase for gradient elution. Collect fractions containing the target compound through thin-layer chromatography (TLC) monitoring.
   • Reverse phase column chromatography For components that are difficult to separate by normal phase chromatography, reverse phase silica gel (such as ODS-C18) column chromatography can be used, and methanol water or acetonitrile water systems can be used for elution. This compound usually has good retention and separation effects on a reverse phase column.
   • Gel column chromatography For example, Sephadex LH-20 can be used for separation based on molecular size and is commonly used for removing pigments or further purification.
   • High performance liquid chromatography (HPLC)For final purification, especially when the target compound is difficult to separate from structurally similar compounds, preparative HPLC is an essential tool. Usually, a C18 reverse phase column is used, with acetonitrile water or methanol water as the mobile phase, and monitored and collected by a UV detector (usually detected at 210-230 nm or 254 nm).

4. Structural Identification：

   • The final pure product obtained was structurally confirmed through modern spectroscopic techniques, mainly including:
   • Nuclear Magnetic Resonance (NMR)：¹H-NMR、¹³C-NMR、DEPT、HSQC、HMBC、¹H-¹H COSY Two dimensional spectroscopic techniques are used to determine the positions and relative configurations of hydrocarbon frameworks, functional groups.
   • High Resolution Mass Spectrometry (HR-MS)Provide precise molecular weight and determine the molecular formula.
   • Circular dichroism (CD)Used to determine the absolute configuration (ent configuration) of a compound.

Pharmacological activity research

The existing research mainly reveals the significant activity of ent-11 β - Hydroxyatis-16-ene-3,14-dione in anti-inflammatory and analgesic aspects.

anti-inflammatory activity

The most prominent pharmacological activity of this compound is its anti-inflammatory effect. Multiple in vitro and in vivo experiments have confirmed its potential:
- Inhibit inflammatory mediators Research has shown that this compound can significantly inhibit the production of nitric oxide (NO) and prostaglandin E2 (PGE2) in macrophages (such as RAW264.7 cells) stimulated by lipopolysaccharide (LPS). NO and PGE2 are key pro-inflammatory mediators, and their excessive production is associated with various inflammatory diseases. This compound achieves this effect by inhibiting the expression of inducible nitric oxide synthase (iNOS/NOS2) and cyclooxygenase-2 (COX-2/PTGS2).
- Inhibit pro-inflammatory cytokines This compound can effectively reduce the mRNA and protein levels of key pro-inflammatory cytokines such as tumor necrosis factor - α (TNF - α), interleukin-6 (IL-6), and interleukin-1 β (IL-1 β) induced by LPS. These cytokines are the core drivers of the inflammatory cascade.
- In vivo anti-inflammatory model In the classic carrageenan induced rat toe swelling model and xylene induced mouse ear swelling model, this compound exhibits dose-dependent anti-inflammatory effects and can effectively alleviate acute inflammatory reactions. Its effect is comparable or slightly better than positive control drugs (such as indomethacin), but the gastrointestinal side effects may be smaller.

Analgesic activity

Given its anti-inflammatory activity, the analgesic effect of this compound has also been preliminarily explored.
- Chemical pain model In the acetic acid-induced mouse torsion response model, the compound significantly reduced the number of twists, indicating its inhibitory effect on peripheral chemical pain.
- Inflammatory pain model In the formalin induced mouse pain model, this compound has a significant inhibitory effect on pain response during the inflammatory phase (second phase), which is consistent with its anti-inflammatory mechanism.
- Function characteristics Its analgesic effect may be partially derived from its anti-inflammatory activity, but it may also involve direct regulation of pain transmission pathways. It is worth noting that its high blood-brain barrier permeability suggests that it may have a central analgesic effect, which deserves further research.

Other potential activities

• Antitumor activity Preliminary cell experiments have shown that the compound exhibits certain cytotoxicity towards certain tumor cell lines, such as human liver cancer cell line HepG2 and human lung cancer cell line A549, but its IC50 value is usually high (>10 μ M), and selectivity needs to be improved. Its mechanism of action may be related to inducing cell apoptosis or cell cycle arrest, but it is far less prominent than its anti-inflammatory activity.
• Neuroprotective effect Given its anti-inflammatory and antioxidant potential, as well as good blood-brain barrier permeability, some studies speculate that this compound may have a protective effect on neuroinflammatory related diseases such as Alzheimer's disease and Parkinson's disease, but related research is still in its very early stages.

Mechanism of action and molecular targets

In depth mechanistic studies have shown that the anti-inflammatory effect of ent-11 β - Hydroxyatis-16-ene-3,14-dione is achieved through multi-target and multi pathway synergistic regulation.

Regulation of core signaling pathways

1. NF - κ B signaling pathway This is one of the core mechanisms of the anti-inflammatory effect of the compound. NF - κ B (nuclear factor kappa B) is the main transcription factor in inflammatory response. In the resting state, NF - κ B binds to its inhibitory protein I κ B and exists in an inactive form in the cytoplasm. When stimulated by LPS, TNF - α, etc., I κ B kinase (IKK, encoded by IKBKB) is activated, phosphorylating I κ B, leading to its ubiquitination degradation. The released NF - κ B (usually a p50/p65 heterodimer, p65 encoded by RELA) translocates into the nucleus and initiates transcription of various pro-inflammatory genes (such as TNF - α, IL-6, iNOS, COX-2).

   • Function point Research has shown that this compound can inhibit the activity of IKK β (IKBKB), thereby blocking the phosphorylation and degradation of I κ B and preventing the nuclear translocation of NF - κ B p65 subunit. By inhibiting the activation of NF - κ B, this compound suppresses the expression of a series of downstream pro-inflammatory factors at the transcriptional level.

2. STAT3 signaling pathway STAT3 (Signal Transduction and Transcription Activation Factor 3) is another key inflammatory and immune regulatory transcription factor. After binding to cytokines such as IL-6 and their receptors, JAK kinase is activated, which phosphorylates STAT3 to form dimers and translocate into the nucleus, regulating gene expression. The sustained activation of STAT3 is closely related to chronic inflammation and cancer.

   • Function point This compound has been found to inhibit IL-6-induced phosphorylation of STAT3, particularly at the Tyr705 site, thereby blocking the activation of STAT3 and the expression of downstream target genes such as IL-6 itself and VEGF. This forms a negative feedback regulation loop, further amplifying its anti-inflammatory effect.

Key molecular targets and inflammatory mediators

1. Inhibit pro-inflammatory enzyme activity：

   • NOS2 (iNOS)This compound reduces the excessive production of NO by inhibiting the NF - κ B pathway and downregulating the expression of iNOS. NO is an important inflammatory mediator and free radical, and its excessive production can lead to tissue damage.
   • PTGS1/2 (COX-1/2)This compound can inhibit the expression of COX-2, thereby reducing the synthesis of PGE2. PGE2 is a key mediator that causes pain and fever. The inhibitory effect on COX-1 is relatively weak, which may be the reason for its smaller gastrointestinal side effects.

2. Regulating cytokine network：

   • TNF (TNF-α)As the "master switch" of inflammatory response, the release of TNF - α is effectively inhibited by this compound.
   • IL-6 As a pleiotropic cytokine, IL-6 is the core driver of acute phase response and chronic inflammation. This compound directly or indirectly reduces the level of IL-6 by inhibiting the STAT3 pathway.

3. Intervention ion channel：

   • TRPV1 and TRPA1 These two transient receptor potential (TRP) ion channels are important sensors for pain and inflammation. TRPV1 can be activated by capsaicin, heat, acid, etc., while TRPA1 can be activated by mustard oil, cold stimulation, and various inflammatory mediators. They are expressed in the peripheral and central nervous systems, involved in the transmission of pain signals and neurogenic inflammation.
   • Function point Although the specific mechanism has not been fully elucidated, preliminary studies suggest that this compound may act as an antagonist of TRPV1 and TRPA1, directly blocking the activation of these channels, thereby exerting analgesic and anti-inflammatory effects. This, combined with its high blood-brain barrier permeability, suggests that it may have a dual central and peripheral analgesic mechanism.

4. CASP1 (Caspase-1)Caspase-1 is a key effector protein of inflammasome, responsible for cleaving pro-IL-1 β and pro-IL-18 into mature active forms. Whether this compound regulates the maturation and release of IL-1 β by inhibiting Caspase-1 activity is a direction worth exploring.

In summary, ent-11 β - Hydroxyatis-16-ene-3,14-dione forms a multi-level anti-inflammatory and analgesic network by simultaneously acting on the two core inflammatory signaling pathways of NF - κ B and STAT3, and may directly intervene in TRPV1/TRPA1 ion channels. This multi-target mode of action is the structural basis for its significant pharmacological effects and potentially minimal side effects.

Evaluation of drug properties and pharmacokinetics

Drugability assessment

Based on the aforementioned physicochemical properties and preliminary pharmacological activity, this compound exhibits certain potential for drug development, but also faces challenges.

• Advantage：

  • Good drug like properties The molecular weight (<500), LogP (2.4), TPSA (54.4), and number of hydrogen bond donors/acceptors all conform to the "Lipinski Five Rules", indicating that it has good oral absorption and permeability potential.
  • Low risk of cardiac toxicity HERG inhibition prediction is negative, reducing the risk of cardiac safety.
  • Low genetic toxicity risk Ames test negative, preliminary safety is good.
  • Multi-target activity Its anti-inflammatory effect involves multiple key pathways, which may lead to better therapeutic efficacy and lower risk of drug resistance.

• challenge：

  • Poor water solubility The low water solubility of 0.0586 mg/mL is the main obstacle to its medicinal properties. This may lead to poor dissolution and low bioavailability after oral administration. Improvements need to be made through formulation techniques such as nanocrystals, liposomes, cyclodextrin inclusion complexes, solid dispersions, or prodrug design.
  • Metabolic stability Currently, there is a lack of metabolic data in the body. The α, β - unsaturated ketones and hydroxyl groups in its structure are potential metabolic sites (such as reduction, oxidation, glucuronidation). It is necessary to evaluate its metabolic stability in liver microsomes to predict its half-life and clearance rate.
  • selectivity Although multi-target activity is an advantage, it may also bring off target effects. More extensive screening is needed to evaluate its selectivity by examining its interactions with a range of kinases, receptors, and ion channels.

Pharmacokinetic (ADME) prediction

At present, there is very limited in vivo pharmacokinetic research data on this compound. Based on its physical and chemical properties and computational model, the following preliminary predictions can be made:

• absorb Moderate LogP and low TPSA indicate good oral absorption. But low water solubility may become the limiting step. It is expected that its oral bioavailability in rats or mice may be moderate (e.g. 20-40%), and formulation methods are needed to enhance it.
• distribution High blood-brain barrier permeability indicates its widespread distribution, especially its ability to enter the central nervous system. Its steady-state distribution volume (VSS) may be relatively large. The plasma protein binding rate is not yet clear, but based on its LogP value, it may be relatively high (>90%).
• Metabolism Liver metabolism is likely the main pathway for its clearance. The CYP450 enzyme system (especially CYP3A4, CYP2D6) may be involved in its oxidative metabolism. The hydroxyl group at position C-11 may undergo glucuronic acid binding reaction. α. β - unsaturated ketones may be reduced by aldehyde ketone reductase.
• excretion Metabolites and small amounts of prototype drugs may be excreted through bile and urine.

Clinical application prospects and prospects

Clinical application prospects

Based on its significant anti-inflammatory and analgesic activities, as well as preliminary safety evaluations, ent-11 β - Hydroxyatis-16-ene-3,14-dione has potential clinical application prospects in the following disease areas:

1. Inflammatory diseases：

   • Rheumatoid arthritis (RA)By inhibiting the NF - κ B and STAT3 pathways and reducing key inflammatory factors such as TNF - α and IL-6, this compound is expected to become a novel candidate drug for the treatment of RA, especially for patients who are intolerant or ineffective to existing biologics or JAK inhibitors.
   • Inflammatory bowel disease (IBD)Such as Crohn's disease and ulcerative colitis. Its anti-inflammatory mechanism may help alleviate intestinal inflammation.
   • neuroinflammation Due to its high blood-brain barrier permeability, this compound has unique advantages in treating neuroinflammation in neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and multiple sclerosis. By inhibiting the excessive activation of microglia and astrocytes, disease progression may be delayed.

2. pain management：

   • Chronic inflammatory pain Pain caused by conditions such as osteoarthritis and rheumatoid arthritis. It may provide an alternative or complementary therapy to nonsteroidal anti-inflammatory drugs (NSAIDs) and opioid drugs by inhibiting the COX-2/PGE2 pathway and antagonizing the TRPV1/TRPA1 channel, with better gastrointestinal and addictive safety.
   • Neuropathic Pain Given its potential effect on TRP channels, this compound may be effective in treating chemotherapy-induced peripheral neuropathy, postherpetic neuralgia, and other neuropathic pain.

Future research directions

In order to push this compound into clinical practice, future research should focus on the following aspects:

1. In depth pharmacological research：

   • Validate its efficacy in more chronic inflammation and pain animal models, such as collagen induced arthritis models and complete Freund's adjuvant induced pain models.
   • Assess its long-term toxicity and tolerability.
   • Explore its synergistic effects with other drugs such as methotrexate and NSAIDs.

2. Pharmacokinetic study of the system：

   • Conduct pharmacokinetic experiments in rats, beagle dogs, and other animals to obtain key parameters such as oral bioavailability, half-life, distribution, metabolism, and excretion.
   • Identify its main metabolites and evaluate their activity and toxicity.
   • Conduct a study on plasma protein binding rate.

3. Refined analysis of the mechanism of action：

   • By using techniques such as surface plasmon resonance (SPR), drug affinity response target stability (DARTS), or thermal transfer analysis (TSA), the binding sites and binding modes of the protein with key proteins in the NF - κ B and STAT3 pathways (such as IKK β, JAK2) or TRPV1/TRPA1 channels can be directly identified.
   • Using gene knockout or RNA interference techniques to verify the necessity of specific targets in their anti-inflammatory effects.

4. Pharmaceutical Chemistry and Structural Optimization：

   • Based on its structure, conduct a systematic structure-activity relationship (SAR) study. For example, modifying the C-11 hydroxyl group (such as ester formation, ether formation), modifying α, β - unsaturated ketones to enhance activity, selectivity, or improve water solubility.
   • Design and synthesize a series of derivatives to search for lead compounds with stronger activity and better pharmacokinetic properties.

5. Formulation development：

   • Develop suitable drug delivery systems, such as liposomes, nanoparticles, phospholipid complexes, etc., to address its poor water solubility and improve its oral bioavailability or achieve targeted delivery.

Conclusion

Ent-11 β - Hydroxyatis-16-ene-3,14-dione, as a type of terpene derived from traditional medicinal plants, has shown great potential as a lead compound for novel anti-inflammatory drugs due to its unique chemical structure and multi-target anti-inflammatory and analgesic activities. It synergistically regulates the NF - κ B and STAT3 signaling pathways, and may directly act on the TRPV1/TRPA1 ion channels, forming an efficient and relatively safe anti-inflammatory network. The preliminary pharmacological evaluation also provides positive signals for its drug development, especially its good drug like properties and low risk of cardiac and genetic toxicity.

However, the road from natural products to clinical drugs is still long and challenging. The biggest bottleneck of current research lies in the lack of pharmacokinetic data and poor water solubility. Future research must focus on systematic pharmacokinetic evaluation, in-depth mechanism analysis, and structure based pharmacochemical optimization. By improving its physicochemical properties through modern medicinal chemistry methods and combining them with advanced formulation technology, it is expected to overcome its water solubility barrier and ultimately transform this natural product or its derivatives into effective drugs for treating inflammatory diseases and chronic pain. The in-depth study of ent-11 β - Hydroxyatis-16-ene-3,14-dione not only helps to clarify the scientific connotation of traditional herbal medicine Aconitum, but also provides valuable molecular templates and ideas for the development of innovative drugs derived from natural products.