(5E,9E,13E)-Teprenone does not have a single defined target but functions primarily by inducing the transcriptional activation of heat shock protein (HSP) genes, particularly HSP70 (also known as HSP72). Through this mechanism, it increases the synthesis and accumulation of cytoprotective HSPs in gastric mucosal cells. HSP70 functions as a molecular chaperone, helping to maintain cellular protein homeostasis, prevent protein aggregation, facilitate protein refolding, and protect cells against a variety of stressors including heat shock, oxidative stress, ischemia, inflammation, and exposure to toxins. The induction of HSP70 is thought to increase gastric mucosal defense under conditions of stress. (5E,9E,13E)-Teprenone may also affect the expression of other HSP family members such as HSP27 and HSP60, and may modulate additional cytoprotective pathways. It is an isomer of teprenone, which is clinically used in Japan and other Asian countries as a gastric mucosal protective agent. The (5E,9E,13E) stereoisomer represents the all-trans geometry at the three double bonds, which is the most biologically active configuration for HSP induction. The compound does not inhibit gastric acid secretion, distinguishing it from other anti-ulcer agents such as histamine H2 receptor antagonists and proton pump inhibitors. Its mechanism is cytoprotective rather than antisecretory.

(5E,9E,13E)-Teprenone has the potential to momentarily stimulate the HSP70 gene's transcription [1]. When (5E,9E,13E)-Teprenone is present, heat shock element binding activity indicates that heat shock factor 1 is activated [1].

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(5E,9E,13E)-Teprenone increases the expression of HSP70 mRNA and protein in cultured gastric mucosal cells. This induction is mediated through the activation of heat shock transcription factor 1 (HSF1), which binds to heat shock elements (HSEs) in the promoter regions of HSP genes. In addition to HSP induction, (5E,9E,13E)-Teprenone enhances the resistance of gastric epithelial cells to injury induced by various stressors, including ethanol, bile salts, and NSAIDs, as demonstrated by increased cell viability and reduced lactate dehydrogenase (LDH) release. The compound also increases the production of gastric mucus and prostaglandins, which contribute to gastric mucosal protection. The detailed in vitro activity beyond these cytoprotective effects is primarily focused on HSP70 upregulation and the consequent protection of cellular proteins against denaturation and aggregation under stress conditions. Quantitative analysis of HSP70 levels is typically performed by Western blotting, ELISA, or RT-PCR.

(5E,9E,13E)-Teprenone causes rats' stomach mucosa to accumulate sock protein [1].

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In vivo studies in rat models have demonstrated that (5E,9E,13E)-Teprenone induces HSP70 accumulation in rat gastric mucosa. Oral administration of the compound (typically 200-600 mg/kg) leads to significant increases in HSP70 protein levels in gastric tissue, as detected by immunohistochemistry, Western blotting, and ELISA. This increase in HSP70 is associated with enhanced gastric mucosal defense against various ulcerogenic stimuli. In standard anti-ulcer models, such as ethanol-induced gastric lesions, water-immersion restraint stress-induced ulcers, and NSAID (e.g., indomethacin)-induced gastric damage, (5E,9E,13E)-Teprenone demonstrates significant protective effects, reducing the number and area of gastric mucosal lesions. The protection is attributed to HSP70-mediated cytoprotection rather than inhibition of gastric acid secretion. The compound has also been shown to prevent the activation of stress-activated protein kinases (SAPK/JNK) and to inhibit apoptosis in gastric mucosal cells. Furthermore, it promotes the healing of existing gastric ulcers by stimulating angiogenesis and granulation tissue formation. The all-trans isomer (5E,9E,13E)-Teprenone is considered the active form, and its efficacy in animal models is well established. Teprenone is clinically used as a gastric mucosal protective agent, and its biological activities have been confirmed in human clinical studies as well.

For in vitro HSP70 induction assays, cultured rat gastric mucosal epithelial cells (RGM-1 cells) or human gastric cancer cell lines (e.g., AGS, MKN-45) are maintained in RPMI-1640 or DMEM medium supplemented with 10% fetal bovine serum and antibiotics. Cells are seeded in 6-well plates (5×10⁵ cells/well) or 25-75 cm2 flasks and allowed to attach overnight. (5E,9E,13E)-Teprenone is added to the culture medium at concentrations ranging from 10-400 microM (typically 100 microM) using DMSO as a vehicle (final DMSO concentration ≤0.1% to avoid cytotoxicity). Control cells receive vehicle alone. Incubation is carried out at 37degC in a 5% CO2 humidified incubator for varying time periods (1-24 hours) to determine the time course of HSP70 induction. For protein analysis by Western blotting: At the end of the treatment period, cells are washed with PBS and lysed in RIPA buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM EDTA) containing protease inhibitor cocktail and phosphatase inhibitors. Lysates are cleared by centrifugation at 12,000 × g for 15 minutes at 4degC. Protein concentration is determined using the BCA assay. Equal amounts of protein (20-50 microg per lane) are separated by SDS-PAGE (10-12% gel) and transferred to PVDF or nitrocellulose membranes. Membranes are blocked with 5% non-fat dry milk or BSA in TBST (Tris-buffered saline with Tween-20) for 1 hour at room temperature, then incubated with primary antibodies against HSP70 (mouse monoclonal or rabbit polyclonal, 1:500-1:2000 dilution) overnight at 4degC. After washing with TBST, membranes are incubated with HRP-conjugated secondary antibodies (1:2000-1:10000) for 1 hour at room temperature. Protein bands are visualized using enhanced chemiluminescence (ECL) substrate, and band intensities are quantified by densitometry using ImageJ or similar software. beta-Actin or GAPDH is used as a loading control. For RNA analysis by RT-PCR: Total RNA is extracted from cells using TRIzol reagent or a commercial RNA isolation kit. cDNA is synthesized using reverse transcriptase. Quantitative real-time PCR is performed using SYBR Green or TaqMan probes with specific primers for HSP70 and housekeeping genes (e.g., GAPDH, beta-actin). Relative HSP70 mRNA expression is calculated using the deltadeltaCt method.

To assess the cytoprotective effect, cells are pre-treated with (5E,9E,13E)-Teprenone for 2-24 hours, then exposed to a cytotoxic stressor such as ethanol (5-15% v/v), indomethacin (500-1000 microM), or H2O2 (100-500 microM) for 4-24 hours. Cell viability is measured by MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) or by quantifying LDH release in the culture supernatant. The percentage of protection is calculated as the viability of stressed cells with pre-treatment relative to stressed cells with vehicle pre-treatment.

Animal/Disease Models: Male Wister rat (approximately 250g) [1]
Doses: 200mg/kg
Route of Administration: Oral
Experimental Results:Causes accumulation of all HSP90, HSP70, HSC70 and HSP60 within 60 minutes.

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For in vivo anti-ulcer studies using rat models, male Sprague-Dawley or Wistar rats (200-250 g, n=8-12 per group) are fasted for 24 hours (free access to water) before the experiment. Several well-established models are used to evaluate the gastroprotective effects of (5E,9E,13E)-Teprenone: Model 1 - Ethanol-induced gastric ulcer: (5E,9E,13E)-Teprenone is administered orally at doses of 100, 200, or 400 mg/kg in a vehicle (e.g., 0.5% carboxymethyl cellulose, CMC, or olive oil). One hour after administration, gastric ulcers are induced by oral administration of absolute ethanol (1 mL per rat). Control animals receive vehicle alone. One hour after ethanol administration, rats are euthanized, and stomachs are removed, opened along the greater curvature, and gently rinsed with saline. The total area (mm2) of hemorrhagic lesions in the glandular part of the stomach is measured under a dissecting microscope. The ulcer index (total lesion area per stomach) is calculated, and the percentage of inhibition is determined relative to the ethanol-only control group. Model 2 - Water-immersion restraint stress-induced ulcer: Rats are orally administered (5E,9E,13E)-Teprenone (100-400 mg/kg) 1 hour before stress exposure. Rats are then placed in a restraint cage (wire mesh) and immersed vertically in a water bath (23 +/- 1degC) to the level of the xiphoid process for 6-8 hours. After the stress period, rats are euthanized, and gastric lesions are quantified as described above. Model 3 - NSAID (indomethacin)-induced gastric ulcer: Rats are orally administered (5E,9E,13E)-Teprenone (200-400 mg/kg) twice daily for 2-3 days. On the final day, indomethacin (30 mg/kg) is administered orally. Rats are euthanized 6 hours later, and gastric lesions are quantified. For histopathological assessment: Gastric tissue samples are fixed in 10% buffered formalin, embedded in paraffin, sectioned (5 microm thickness), and stained with hematoxylin and eosin (H&E). Sections are examined under a light microscope for morphological changes (epithelial cell loss, edema, inflammatory cell infiltration). For immunohistochemistry of HSP70: Paraffin-embedded gastric tissue sections are deparaffinized, rehydrated, and subjected to antigen retrieval (e.g., microwave heating in citrate buffer). Sections are incubated with anti-HSP70 primary antibody (overnight at 4degC), followed by HRP-conjugated secondary antibody. Staining is visualized using diaminobenzidine (DAB) chromogen, and sections are counterstained with hematoxylin. The intensity and distribution of HSP70 immunoreactivity are scored (semi-quantitative) or quantified by image analysis. For Western blotting of HSP70 in gastric tissue: Gastric mucosal samples are scraped, homogenized in RIPA buffer, and processed for Western blot analysis as described for the in vitro section. For assessment of gastric mucus production: Gastric tissue sections are stained with periodic acid-Schiff (PAS) to visualize mucopolysaccharides, and the thickness of the mucus layer or the area of PAS-positive staining is measured. For measurement of gastric prostaglandin E2 (PGE2) levels: Gastric mucosal homogenates are prepared, and PGE2 concentration is measured using a commercial ELISA kit. For gastric acid secretion studies: Rats are pylorus-ligated under anesthesia (Shay rat model). (5E,9E,13E)-Teprenone or vehicle is administered orally immediately after pylorus ligation. Four hours later, the gastric contents are collected, and the volume, pH, and titratable acidity are measured. As expected, (5E,9E,13E)-Teprenone does not significantly affect acid secretion, distinguishing it from antisecretory anti-ulcer drugs.

As a synthetic polyisoprenoid compound used orally, teprenone is known to be well-absorbed from the gastrointestinal tract. Clinical formulations of teprenone (e.g., Selbex, Geranylgeranylacetone) are available in Japan and other Asian countries, indicating that the compound has acceptable absorption, distribution, metabolism, and excretion (ADME) properties in humans. The compound is lipophilic due to its polyisoprenoid structure, allowing it to penetrate biological membranes, including the gastric mucosal epithelium. Following oral absorption, teprenone is distributed to the gastric mucosa and other tissues, where it induces HSP70 expression. The compound is metabolized in the liver, likely via oxidation of the terminal isoprenoid units, forming various polar metabolites. Detailed pharmacokinetic parameters (e.g., plasma half-life, Cmax, Tmax, AUC, volume of distribution, protein binding) for the all-trans isomer (5E,9E,13E)-teprenone specifically are not fully detailed in supplier datasheets, as the compound is primarily used as a research standard and the parent compound teprenone (a mixture of isomers or the unspecified isomer) is the clinical drug. The molecular weight of (5E,9E,13E)-Teprenone is 330.55, and its molecular formula is C23H3₈O. It is typically stored as a powder at -20degC or as a 10 mM solution in DMSO (ready-to-use). The compound is soluble in DMSO and organic solvents, and is also formulated as an oil or suspension for oral administration in preclinical studies.

(5E,9E,13E)-Teprenone is generally considered to have a favorable safety profile, consistent with its clinical use as a gastric mucosal protective agent. Teprenone has been approved for clinical use in Japan (Selbex) and several other Asian countries for the treatment of gastric ulcers, acute gastritis, and chronic gastritis, indicating that it is well-tolerated in humans. Common adverse effects reported in clinical practice are mild and may include gastrointestinal discomfort, diarrhea, constipation, or nausea, but serious toxicity is rare. In preclinical animal studies, (5E,9E,13E)-Teprenone has not shown significant acute toxicity at doses up to several thousand mg/kg orally. The compound does not inhibit COX enzymes (unlike NSAIDs) and does not cause gastric mucosal injury. Long-term safety studies have not raised major concerns for the parent drug teprenone. Formal Good Laboratory Practice (GLP) toxicity studies for the isolated isomer may be limited, but given that teprenone products are marketed, the overall safety of this class of compounds is well established for oral use. The compound is intended for research use only in its pure isomer form, and users should refer to the Material Safety Data Sheet (MSDS) for specific handling precautions. As with any lipophilic compound, acute ingestion of very large doses could potentially cause gastrointestinal irritation. However, at research-relevant concentrations and doses (uM in vitro, mg/kg in vivo), (5E,9E,13E)-Teprenone is considered to have a low toxicity profile. It is not known to be genotoxic, carcinogenic, or teratogenic at therapeutic doses, but dedicated studies for the isolated isomer may not be available.

[1]. Geranylgeranylacetone induces heat shock proteins in cultured guinea pig gastric mucosal cells and rat gastric mucosa. Gastroenterology. 1996 Aug;111(2):345-57.

Teprenone is a terpenoid ketone in which a (9E,13E)-geranyl-geranyl group is linked to the α-methyl group of acetone (it is a mixture of 5E- and 5Z-geometric isomers in a 3:2 ratio). It possesses anti-ulcer, cardioprotective, hepatoprotective, nephroprotective, neuroprotective, and Hsp70-inducing effects. It is a methyl ketone and a terpenoid ketone containing a geranyl-geranyl group.

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Mechanism of Action
Teprenone is an anti-ulcer and gastric mucosal protectant used to treat gastric ulcers and gastritis. Currently, its complete mechanism of action is not fully understood.

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(5E,9E,13E)-Teprenone (CAS# 3796-63-2) is a research-grade isomer of teprenone, also known as (5E,9E,13E)-Geranylgeranylacetone or all-trans geranylgeranylacetone. It has a molecular weight of 330.55 and a molecular formula of C23H3₈O. Teprenone is an anti-ulcer agent that works by inducing the expression of cytoprotective heat shock proteins (especially HSP70) in the gastric mucosa, a mechanism distinct from acid suppression. The compound is clinically used as a gastric mucosal protective agent in Japan (trade name Selbex) and other Asian countries for the treatment of gastric ulcers, duodenal ulcers, acute gastritis, and chronic gastritis. It is also used to prevent gastric mucosal injury induced by nonsteroidal anti-inflammatory drugs (NSAIDs) and stress. The all-trans isomer (5E,9E,13E)-Teprenone is the biologically active configuration. This compound is employed in biochemical and pharmacological research to study the molecular mechanisms of heat shock protein induction (HSF1 activation, HSE binding), the role of HSP70 in cytoprotection against oxidative stress, inflammation, and apoptosis, and to evaluate potential HSP70 inducers for various therapeutic indications, including neurodegenerative diseases (where HSP70 induction may prevent protein aggregation) and ischemia-reperfusion injury (cardioprotection, neuroprotection). (5E,9E,13E)-Teprenone is not approved for clinical use in the United States or Europe but is an important research standard for studies related to HSP biology and mucosal protection. It is typically stored at -20degC as a solid or as a 10 mM solution in DMSO, and is soluble in organic solvents. The compound is also used as a reference material for analytical method development (HPLC, GC, LC-MS) for the quantification of teprenone and its isomers in pharmaceutical formulations and biological samples. For researchers aiming to study teprenone‘s pharmacology, this specific isomer provides a well-defined chemical entity for reproducible experiments. The compound should not be used for human consumption and is intended for laboratory research only.

C₂₃H₃₈O

330.55

660.585

3796-63-2

Teprenone;6809-52-5

5282199

Colorless to light yellow liquid

0.871g/cm3

438.2ºC at 760 mmHg

166.5ºC

1.482

15.002

0

1

12

24

482

0

CC(=CCC/C(=C/CC/C(=C/CC/C(=C/CCC(=O)C)/C)/C)/C)C

HUCXKZBETONXFO-NJFMWZAGSA-N

InChI=1S/C23H38O/c1-19(2)11-7-12-20(3)13-8-14-21(4)15-9-16-22(5)17-10-18-23(6)24/h11,13,15,17H,7-10,12,14,16,18H2,1-6H3/b20-13+,21-15+,22-17+

(5E,9E,13E)-6,10,14,18-tetramethylnonadeca-5,9,13,17-tetraen-2-one

(5E,9E,13E)Teprenone; (5E,9E,13E) Teprenone

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~5 mg/mL (~15.13 mM)

Solubility in Formulation 1: ≥ 0.5 mg/mL (1.51 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 5.0 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: ≥ 0.5 mg/mL (1.51 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 5.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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