Hubbry Logo
ThapsigarginThapsigarginMain
Open search
Thapsigargin
Community hub
Thapsigargin
logo
8 pages, 0 posts
0 subscribers
Be the first to start a discussion here.
Be the first to start a discussion here.
Thapsigargin
Thapsigargin
Thapsigargin
View on Wikipedia
from Wikipedia

Thapsigargin
Thapsigargin
Thapsigargin
Names
IUPAC name
(11S)-7,11-Dihydroxy-12-oxo-6β,12-epoxy-1β,7α,10α-guai-4-ene-2β,3α,8α,10-tetrayl 10-acetate 8-butanoate 3-[(2Z)-2-methylbut-2-enoate] 2-octanoate
Systematic IUPAC name
(3S,3aR,4S,6S,6aR,7S,8S,9bS)-3,3a-Dihydroxy-3,6,9-trimethyl-2-oxo-2,3,3a,4,5,6,6a,7,8,9b-decahydroazuleno[4,5-b]furan-4,6,7,8-tetrayl 6-acetate 4-butanoate 8-[(2Z)-2-methylbut-2-enoate] 7-octanoate
Identifiers
3D model (JSmol)
ChEBI
ChEMBL
ChemSpider
ECHA InfoCard 100.116.539 Edit this at Wikidata
UNII
  • InChI=1S/C34H50O12/c1-9-12-13-14-15-17-24(37)43-28-26-25(20(5)27(28)44-30(38)19(4)11-3)29-34(41,33(8,40)31(39)45-29)22(42-23(36)16-10-2)18-32(26,7)46-21(6)35/h11,22,26-29,40-41H,9-10,12-18H2,1-8H3/b19-11-/t22-,26+,27-,28-,29-,32-,33+,34+/m0/s1 checkY
    Key: IXFPJGBNCFXKPI-FSIHEZPISA-N checkY
  • InChI=1/C34H50O12/c1-9-12-13-14-15-17-24(37)43-28-26-25(20(5)27(28)44-30(38)19(4)11-3)29-34(41,33(8,40)31(39)45-29)22(42-23(36)16-10-2)18-32(26,7)46-21(6)35/h11,22,26-29,40-41H,9-10,12-18H2,1-8H3/b19-11-/t22-,26+,27-,28-,29-,32-,33+,34+/m0/s1
    Key: IXFPJGBNCFXKPI-FSIHEZPIBR
  • O=C3O[C@H]2C\1=C(\[C@H](OC(=O)\C(=C/C)C)[C@@H](OC(=O)CCCCCCC)[C@@H]/1[C@@](OC(=O)C)(C[C@H](OC(=O)CCC)[C@]2(O)[C@@]3(O)C)C)C
Properties
C34H50O12
Molar mass 650.762 g·mol−1
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
☒N verify (what is checkY☒N ?)

Thapsigargin is a non-competitive inhibitor of the sarco/endoplasmic reticulum Ca2+ ATPase (SERCA).[1] Structurally, thapsigargin is classified as a guaianolide, and is extracted from a plant, Thapsia garganica.[2] It is a tumor promoter in mammalian cells.[3]

Thapsigargin raises cytosolic (intracellular) calcium concentration by blocking the ability of the cell to pump calcium into the sarcoplasmic and endoplasmic reticula. Store-depletion can secondarily activate plasma membrane calcium channels, allowing an influx of calcium into the cytosol. Depletion of ER calcium stores leads to ER stress and activation of the unfolded protein response.[4] Non-resolved ER stress can cumulatively lead to cell death.[5][6] Prolonged store depletion can protect against ferroptosis via remodeling of ER-synthesized phospholipids.[7]

Thapsigargin treatment and the resulting ER calcium depletion inhibits autophagy independent of the UPR.[8][9]

Thapsigargin is useful in experimentation examining the impacts of increasing cytosolic calcium concentrations and ER calcium depletion.

A study from the University of Nottingham showed promising results for its use against Covid-19 and other coronavirus.

Biosynthesis

[edit]

The complete biosynthesis of thapsigargin has yet to be elucidated. A proposed biosynthesis starts with the farnesyl pyrophosphate. The first step is controlled by the enzyme germacrene B synthase. In the second step, the C(8) position is easily activated for an allylic oxidation due to the position of the double bond. The next step is the addition of the acyloxy moiety by a P450 acetyltransferase; which is a well known reaction for the synthesis of the diterpene, taxol. In the third step, the lactone ring is formed by a cytochrome P450 enzyme using NADP+. With the butyloxy group on the C(8), the formation will only generate the 6,12-lactone ring. The fourth step is an epoxidation that initiates the last step of the base guaianolide formation. In the fifth step, a P450 enzyme closes the 5 + 7 guaianolide structure. The ring closing is important, because it will proceed via 1,10 - epoxidation in order to retain the 4,5 - double bond needed in thapsigargin. It is not known whether the secondary modifications to the guaianolide occur before, or after the formation of thapsigargin, but will need to be considered when elucidating the true biosynthesis. It should also be noted, that several of these enzymes are P450s, therefore oxygen and NADPH are likely crucial to this biosynthesis as well as other cofactors such as Mg2+ and Mn2+ may be needed.[10]

Research

[edit]

Since inhibition of SERCA is a mechanism of action that has been used to target solid tumors, thapsigargin has attracted research interest. A prodrug of thapsigargin, mipsagargin, is currently undergoing clinical trials for the treatment of glioblastoma.[11][12][13][14]

The biological activity has also attracted research into the laboratory synthesis of thapsigargin. To date, three distinct syntheses have been reported: one by Steven V. Ley,[15] one by Phil Baran.,[16] and one by P. Andrew Evans.[17]

Preclinical studies demonstrated that other effects of thapsigargin include suppression of nicotinic acetylcholine receptors activity in neurons of the guinea-pig ileum submucous plexus[18] and rat superior cervical ganglion.[19]

Laboratory studies at the University of Nottingham, using in vitro cell cultures, indicates possible potential as a broad spectrum antiviral, with activity against the COVID-19 virus (SARS-CoV-2), a common cold virus, respiratory syncytial virus (RSV), and the influenza A virus.[20]

See also

[edit]

References

[edit]

Further reading

[edit]
Revisions and contributorsEdit on WikipediaRead on Wikipedia

Thapsigargin

View on Grokipedia
from Grokipedia
Thapsigargin is a guaianolide isolated from the roots of the Mediterranean plant Thapsia garganica ( family), a species historically known as the "deadly " for its irritant properties. First isolated in 1978 with its structure elucidated in 1985, it features a complex tricyclic framework consisting of a seven-membered ring fused to a five-membered and a γ-lactone ring. As a potent and irreversible inhibitor of the sarco/ Ca²⁺-ATPase (SERCA) pump at subnanomolar to low nanomolar concentrations, thapsigargin depletes calcium stores in the (ER), disrupting calcium homeostasis and triggering ER stress. This mechanism activates the unfolded protein response (UPR) and can induce in various cell types, making thapsigargin a widely used tool in research to study , ER stress pathways, and related cellular processes. In experimental settings, it is typically applied at concentrations of 0.05–1 μM to elicit these effects without immediate , though higher doses (e.g., low micromolar) reveal its potent cytotoxic potential. Historically, extracts from Thapsia garganica have been employed in folk medicine across the Mediterranean for treating rheumatic pain, lung diseases, and , leveraging its and irritant qualities. Beyond research applications, thapsigargin exhibits promising therapeutic potential, particularly as an anticancer agent due to its ability to selectively induce in tumor cells via inhibition and NOTCH1 pathway disruption in certain leukemias. Prodrugs such as mipsagargin (G202), which is activated by prostate-specific membrane antigen (PSMA), have advanced to Phase II clinical trials as of 2025 for , hepatocellular, , and cancers, demonstrating tolerability in Phase I studies and efficacy in preclinical xenograft models. Additionally, thapsigargin displays broad-spectrum antiviral activity against major human respiratory viruses, including , , (RSV), and influenza A, by inducing ER stress and host responses that inhibit viral replication at early stages (e.g., transcription for RSV and OC43) with selectivity indices exceeding 700 in cell cultures; recent 2024 studies confirm activity against additional coronaviruses and noroviruses. Its has been achieved through multi-step processes, with scalable methods reported in 2016 enabling further derivatization for improved specificity and reduced toxicity in clinical contexts.

Overview and History

Discovery and Isolation

Thapsigargin was first isolated in 1978 from the roots of the Mediterranean umbellifer Thapsia garganica L. by researchers at the , including Søren Brøgger Christensen, U. Rasmussen, and F. Sandberg, as part of efforts to characterize the plant's skin-irritant principles. The isolation process involved extracting the ground roots with , followed by concentration and purification of the active components via (HPLC) to yield thapsigargin as a colorless oil. This work identified thapsigargin alongside a related compound, thapsigargicin, both responsible for the irritant effects observed in traditional uses of the plant.98451-3) In the same 1978 study, the initial pharmacological effects of thapsigargin were reported, revealing its potent ability to liberate from isolated rat peritoneal mast cells in a calcium-dependent manner, with an ED50 value around 40 nM. This histamine-releasing activity in mammalian cells underscored its irritant potential and laid the groundwork for further investigations into its biological actions. Subsequent studies in the early confirmed tumor-promoting activity in two-stage mouse skin models, where thapsigargin enhanced formation at doses of 1–10 nmol per application. The full molecular structure of thapsigargin was elucidated in 1985 by Christensen and Norup through a combination of (NMR) spectroscopy and of its 3-O-benzoyl derivative, establishing it as a guaianolide with a unique hexaoxygenated framework and specific at eight chiral centers.98451-3) This structural determination resolved earlier ambiguities from partial degradations and spectroscopic data, confirming thapsigargin's classification within the family and enabling subsequent synthetic and mechanistic studies.

Traditional and Early Uses

Thapsia garganica, the primary source of thapsigargin, has been utilized in ancient Mediterranean folk medicine since at least the time of and around 400–300 BCE, where its resin was applied as a counterirritant for treating , , and other skin conditions, as well as serving as a purgative for digestive ailments. In Greek and Roman traditions, the plant's roots and resin were employed externally to alleviate pain and internally to promote , though its acrid properties often led to severe blistering and upon contact. North African and Arabian healers continued these practices into later centuries, using crude extracts for rheumatic pain relief, lung diseases like , , and fever, despite awareness of its potent irritant effects causing skin and gastrointestinal distress. All traditional applications relied on unrefined extracts, as purified thapsigargin was not isolated until 1978. The plant earned the moniker "deadly carrot" due to its resemblance to edible carrots and its high toxicity, with historical accounts documenting livestock poisoning across North Africa and southern Europe, where ingestion by sheep, cattle, and camels caused rapid onset of gastrointestinal hemorrhage, convulsions, and death. In regions like and , pastoral communities reported fatal cases in grazing animals, attributing the dangers to the resinous sap, which induced severe mucosal irritation and systemic effects even in small amounts. These poisonous properties were well-known in ethnobotanical records from the early , reinforcing caution in human uses and highlighting the plant's dual role as both remedy and . By the early , European botanists and pharmacologists began systematically recognizing the irritant compounds in Thapsia species, noting the presence of sesquiterpenoid lactones responsible for and toxicity, which spurred targeted investigations in the into the bioactive guaianolides within the . These efforts built on folk knowledge but focused on chemical characterization without isolating thapsigargin itself until later. The skin-irritant effects observed in traditional applications have since been linked to disruptions in cellular calcium .

Chemical Properties

Molecular Structure

Thapsigargin is classified as a hexa-oxygenated guaianolide with a 5-7-5 core. The features a 6β,12- bridge, 7α,11-dihydroxy groups, a 12-oxo moiety, and four side chains at positions 2, 3, 8, and 10, comprising a 10-acetate, 8-butanoate, 3-(2Z-2-methylbut-2-enoate), and 2-octanoate. These structural elements define its guaianolide scaffold, with the epoxy bridge and hydroxy groups contributing to the trans-fused ring configuration. The IUPAC name for thapsigargin is (3S,3aR,4S,6S,6aR,7S,8S,9bS)-6-(acetyloxy)-4-(butanoyloxy)-3,3a-dihydroxy-3,6,9-trimethyl-8-{[(2Z)-2-methylbut-2-enoyl]oxy}-9-(octanoyloxy)-3a,4,5,5a,6,6a,7,8-octahydroazuleno[4,5-b]furan-2(3H)-one. Thapsigargin exhibits specific at chiral centers C1β, C7α, C10α, and C11S, which is essential for its biological potency through precise interactions with target proteins.

Physicochemical Characteristics

Thapsigargin possesses the molecular formula \ceC34H50O12\ce{C34H50O12} and a of 650.762 g/mol. At , the compound typically presents as a white powder or a colorless , reflecting its nature. These physical attributes facilitate its handling in laboratory settings, where it is often stored as a solid under desiccated conditions at -20°C to maintain integrity. Solubility profiles of thapsigargin highlight its lipophilic character, with high solubility in organic solvents such as DMSO (≥65 mg/mL), ethanol (≥20 mg/mL), and acetone. In contrast, it exhibits poor solubility in water, requiring ultrasonication to achieve concentrations of approximately 4 mg/mL. This behavior is quantified by its octanol-water partition coefficient (logP) of 4.9, underscoring its preference for non-aqueous environments and aiding its membrane permeability in biological systems. The compound demonstrates sensitivity to light exposure and basic conditions, which can lead to degradation, while remaining stable in neutral pH environments typical of physiological media. Due to its amorphous form, thapsigargin lacks a distinct melting point but undergoes decomposition above 200°C. In terms of inhibitory kinetics, it functions as a non-competitive inhibitor of the sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA) with an IC50 of approximately 0.4 nM.

Natural Occurrence and Biosynthesis

Plant Sources

Thapsigargin is primarily sourced from the roots of Thapsia garganica L., a perennial herbaceous plant in the family native to Mediterranean regions, including , , and . This species thrives in dry, rocky, and stony habitats, such as open shrubby vegetation, fallow fields, groves, and steppe-like areas at low altitudes up to 900 meters, preferring well-drained sandy or loamy soils in sunny positions. The center of its diversity lies in the western Mediterranean, where it grows in arid environments but faces threats from habitat loss and overharvesting due to increasing demand for thapsigargin in research and potential pharmaceutical applications. In T. garganica, thapsigargin concentrations reach 0.2%–1.2% of the dry root weight, with higher levels (0.7%–1.5%) in ripe fruits, though remain the main commercial source. Lower amounts of thapsigargin or related lactones occur in of certain other Thapsia , such as T. transtagana, but T. garganica is the predominant natural producer. Extraction typically begins with drying the , followed by solvent extraction using or 70% to obtain crude extracts, which are then concentrated and purified via chromatography techniques like (HPLC) or centrifugal on to isolate thapsigargin as a pure, oily residue. Wild collection poses sustainability challenges, as T. garganica populations are vulnerable to depletion from unregulated harvesting, prompting conservation efforts including in vitro propagation and greenhouse cultivation in Denmark to provide a stable research supply without relying on wild sources.

Biosynthetic Pathway

The biosynthesis of thapsigargin in Thapsia garganica begins with (FPP), a universal sesquiterpenoid precursor derived from the , which undergoes cyclization to form the guaiane skeleton. The initial committed step is catalyzed by the terpene synthase TgTPS2, which converts FPP to epi-kunzeaol (also referred to as kunzeaol in some contexts), a bicyclic alcohol featuring a 6β-hydroxygermacra-1(10),4-diene structure. This enzyme, identified through sequencing of T. garganica roots, operates in the and requires divalent metal ions such as Mg²⁺ or Mn²⁺ as cofactors to facilitate the ionization and cyclization of FPP, potentially involving allylic rearrangements to establish the essential for downstream modifications. Subsequent transformations involve multiple cytochrome P450-mediated oxidations to introduce functional groups and form the characteristic octahydroazulene core with its fused lactone ring. A key enzyme, CYP76AE2, performs a triple oxidation at the C-12 position of epi-kunzeaol, yielding epi-dihydrocostunolide and initiating lactone ring closure between C-6 and C-12; this P450 is localized in the epithelial cells lining the secretory ducts of roots, where thapsigargin accumulates. Further steps include epoxidation (e.g., at the 6,7-double bond), hydroxylations at positions such as C-7, C-8, C-10, and C-11, and acylation of hydroxyl groups with short-chain fatty acids like acetate and butanoate, likely mediated by acyltransferases. These oxidative and conjugative reactions depend on NADPH and molecular oxygen as cofactors, with an estimated 10-15 enzymatic steps in total, including additional allylic oxidations and rearrangements to achieve the complex guaianolide architecture. The pathway remains hypothetical and incompletely elucidated, with gene clusters partially identified through root transcriptomics but lacking full genomic context due to the absence of a complete T. garganica . is confined to specialized oil secretory structures in and fruits, contributing to low natural yields (typically <1% dry weight), which complicates industrial production. As of 2025, no complete in vitro reconstruction of the pathway has been achieved, hindering metabolic engineering efforts despite identification of early enzymes like TgTPS2 and CYP76AE2.

Mechanism of Action

Molecular Target

Thapsigargin's primary molecular target is the (SERCA) pump family, which actively transports Ca²⁺ ions into the sarcoplasmic reticulum (SR) in muscle cells or the endoplasmic reticulum (ER) in non-muscle cells to maintain intracellular Ca²⁺ homeostasis. It exhibits high potency against the SERCA2b isoform, predominant in the ER of non-muscle cells, with lower potency against SERCA3, which is expressed in specific tissues such as endothelial and epithelial cells. Thapsigargin inhibits all SERCA isoforms but with varying affinities, blocking Ca²⁺ uptake into the ER/SR without affecting Ca²⁺ release channels such as inositol 1,4,5-trisphosphate receptors (IP₃Rs). The inhibition is non-competitive with respect to both Ca²⁺ and ATP, occurring through irreversible binding to a hydrophobic pocket in the transmembrane domain of SERCA, primarily involving helices M3, M5, and M7. This binding stabilizes the enzyme in its Ca²⁺-free E2 conformation, preventing the necessary structural transition to the Ca²⁺-bound E1 state required for ATP hydrolysis and Ca²⁺ translocation across the membrane. The interaction is rapid and stoichiometric, leading to prolonged disruption of ATP-driven Ca²⁺ pumping. Thapsigargin displays subnanomolar affinity for , with reported IC₅₀ values ranging from approximately 0.1 to 2 nM across isoforms, such as 0.35 nM for SERCA1a and around 10 nM for SERCA2b. Structure-activity relationship studies highlight the essential role of specific functional groups in thapsigargin's guaianolide sesquiterpene scaffold for this potency: the 7,11-epoxy bridge and the ester moieties at positions O-2, O-3, O-8, and O-10 are critical, with modifications to the epoxy group or O-2 ester dramatically reducing inhibitory activity against . These elements facilitate tight hydrophobic and hydrogen-bonding interactions within the binding pocket. At higher concentrations (micromolar range), thapsigargin exhibits weak off-target inhibition of other Ca²⁺-ATPases, such as the plasma membrane Ca²⁺-ATPase (PMCA), though with substantially lower potency than for SERCA. Additionally, in neuronal contexts, thapsigargin suppresses activity of nicotinic acetylcholine receptors (nAChRs) indirectly through SERCA-mediated Ca²⁺ store depletion, which dysregulates Ca²⁺-dependent signaling and reduces receptor currents.

Cellular and Physiological Effects

Thapsigargin inhibits the sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA), leading to rapid depletion of Ca²⁺ from endoplasmic reticulum (ER) stores and a subsequent increase in cytosolic Ca²⁺ concentrations. This depletion activates store-operated Ca²⁺ entry (SOCE) through STIM1/Orai1-mediated pathways, as well as store-independent mechanisms involving TRPC3/PLC/PKC signaling in endothelial cells. The resulting Ca²⁺ dysregulation triggers ER stress, activating the unfolded protein response (UPR) via the PERK, IRE1, and ATF6 pathways, which upregulate chaperones like CHOP and promote pro-apoptotic signaling. Independently of UPR activation, thapsigargin inhibits autophagic flux by blocking autophagosome-lysosome fusion via disruption of RAB7 recruitment to autophagosomes, leading to accumulation of autophagosomes without affecting their formation. Calpain activation contributes to early autophagy induction. At the cellular level, these effects culminate in cell death, primarily through apoptosis involving caspase activation (e.g., caspase-8 via DR5 and CHOP), reactive oxygen species (ROS) production, and mitochondrial dysfunction. Thapsigargin can also induce non-apoptotic pathways, such as autosis—a Na⁺/K⁺-ATPase-dependent form of programmed cell death characterized by perinuclear ballooning and autophagolysosome accumulation—in basophilic leukemia cells. Physiologically, thapsigargin causes skin irritation through neurogenic inflammation, stimulating arachidonic acid release and PGE₂ production in keratinocytes, though with lower potency than phorbol esters. It promotes tumor formation in mouse skin models by inducing hyperplasia via regenerative proliferation following cytotoxicity, acting as a weak tumor promoter. In neurons, thapsigargin suppresses nicotinic acetylcholine receptor (nAChR) signaling by elevating intracellular Ca²⁺, inhibiting acetylcholine-induced currents in submucous plexus neurons. The effects are dose-dependent: low nanomolar concentrations (e.g., 1–10 nM) are used for Ca²⁺ signaling and ER stress studies without overt cytotoxicity, while higher micromolar doses (e.g., >1 μM) drive robust .

Research and Applications

Cancer Therapeutics

Thapsigargin's potential as an anticancer agent stems from its inhibition of the sarco/ Ca²⁺-ATPase () pump, which disrupts and induces (ER) stress preferentially in rapidly dividing cancer cells that exhibit elevated demands. This ER stress triggers the unfolded protein response (UPR), leading to via pathways such as JNK activation and CHOP upregulation, while offering a mechanism to circumvent resistance to conventional chemotherapies that rely on damage or disruption. Preclinical studies have demonstrated efficacy against various solid tumors, including , , and hepatocellular carcinomas, where thapsigargin promotes immunogenic by exposing on the cell surface, potentially enhancing antitumor immune responses.02085-2/fulltext) To address thapsigargin's systemic toxicity, researchers have developed that mask its activity until activated in the . The lead compound, mipsagargin (G-202), is a peptide-conjugated cleaved by prostate-specific membrane antigen (PSMA), enabling targeted release in PSMA-expressing cancers like tumors. Phase I trials established its tolerability and in patients with advanced solid tumors, including , with a recommended dose of 3.5 mg/m² administered intravenously over three days in 28-day cycles. Subsequent Phase II studies evaluated mipsagargin in recurrent and sorafenib-refractory , showing disease stabilization in subsets of patients but limited overall response rates, leading to trial completion without further advancement by 2025 due to insufficient efficacy signals. Structure-activity relationship (SAR) studies have guided the synthesis of less toxic thapsigargin analogs by modifying ester groups to create inactive proforms activated by tumor-specific enzymes or conditions, reducing off-target effects while retaining inhibitory potency. These derivatives, when combined with chemotherapies like or , synergistically enhance in preclinical models of esophageal and cancers by amplifying ER stress and overcoming survival signaling. Despite promising preclinical data, challenges persist, including dose-limiting toxicities such as vascular leak syndrome and no FDA approvals to date. Ongoing research focuses on advanced targeted delivery systems, such as encapsulation, to improve tumor specificity and minimize systemic exposure, with preclinical evidence of enhanced cell death via sustained ER stress induction. Analogs incorporating tumor-homing peptides or pH-sensitive linkers continue to advance in early-stage development, aiming to revive thapsigargin's therapeutic potential in .

Antiviral and Other Uses

Thapsigargin has demonstrated broad-spectrum antiviral activity against major respiratory viruses, including , (RSV), and , primarily through induction of (ER) stress via the unfolded protein response (UPR). This host-directed mechanism disrupts viral replication by blocking transcription in coronaviruses and RSV, while inhibiting post-translational processes in A, with observed at low non-cytotoxic nanomolar concentrations (typically 50-500 nM ). Studies from the have highlighted its potential as a host-centric , showing up to 10,000-fold reduction in RSV progeny and significant protection in models of at oral doses of 1.5 μg/kg/day. As of 2025, thapsigargin remains in preclinical stages for antiviral applications, with no approved formulations, but its multimodal action offers promise for rapid deployment against emerging epidemics.00084-0) Beyond antivirals, thapsigargin serves as a key research tool in , particularly for investigating Ca²⁺ signaling and ER stress pathways. It selectively inhibits the sarco/ Ca²⁺-ATPase (), depleting ER Ca²⁺ stores and activating store-operated Ca²⁺ entry (also known as capacitative Ca²⁺ entry), which has been instrumental in defining these mechanisms across various cell types. Its ability to induce UPR without immediate at low doses makes it a standard for modeling ER stress-related processes in neuronal and other cells. Advancements in thapsigargin synthesis have enabled production of analogs for research and potential therapeutics, overcoming limited natural supply from Thapsia garganica. The Ley group achieved the first in 2007, a 42-step enantioselective route starting from (S)- that established access to the core guaianolide scaffold. In 2016, the Baran laboratory reported a more concise 11-step scalable synthesis from (+)-dihydrocarvone, yielding 0.137% overall and supporting gram-scale production of intermediates for analog diversification. Complementing these, the Evans group developed an efficient route in 2017 to nortrilobolide and thapsigargin from (R)-(-)- in fewer steps, emphasizing stereocontrolled assembly for practical analog synthesis. Thapsigargin also shows exploratory potential in neurodegenerative diseases through ER stress modulation, as nanoparticle formulations have protected against amyloid-β-induced in Alzheimer's models by activating UPR pathways. In plant biology, studies reveal its role in , with production induced in Thapsia garganica following simulated herbivory, altering profiles to deter herbivores via sesquiterpene lactone .

Toxicity and Safety

Toxicological Effects

Thapsigargin exhibits high in animal models, with a reported LD50 of 2 mg/kg via in mice, indicating potential for rapid systemic effects following exposure. This toxicity arises primarily from its inhibition of the sarco/ Ca²⁺-ATPase (), leading to (ER) stress and subsequent cellular dysfunction, particularly in calcium-sensitive tissues. In vivo studies demonstrate that thapsigargin administration (1 mg/kg, intraperitoneal) in mice induces ER stress-mediated contractile dysfunction and interstitial in cardiac tissue, underscoring its role in precipitating organ-level damage through and . At the cellular level, thapsigargin is highly cytotoxic, primarily triggering through ER stress and the unfolded protein response, as observed in various cell lines including hepatocytes and cells. It also induces autosis, a non-apoptotic form of characterized by vacuolization and dependence on sodium/potassium activity, particularly at concentrations that deplete ER calcium stores. Sublethal doses promote tumor formation by stimulating in , acting as a weak skin tumor promoter without direct . In animal models, thapsigargin causes significant adverse effects, including skin vesication and irritation upon contact, which historically contributed to the naming of its source plant, Thapsia garganica, after reports of severe burns and blisters from resin exposure. Systemic exposure leads to , manifesting as behavioral impairments linked to ER stress in brain tissue, though direct blockade of nicotinic acetylcholine receptors (nAChRs) has not been confirmed as the primary mechanism. Human data on thapsigargin exposure are limited, with no reported cases of direct systemic intoxication; however, handling the source plant Thapsia garganica commonly results in , presenting as , itching, and small blisters due to the compound's irritant properties. Its tumor-promoting activity raises concerns of potential carcinogenicity, particularly with chronic low-level exposure, though it lacks inherent mutagenicity. Chronic exposure to thapsigargin exacerbates ER stress, leading to lipid accumulation in hepatocytes and other cells via impaired lipid metabolism and unfolded protein response dysregulation. Prolonged effects include fibrosis in susceptible organs, such as cardiac interstitial fibrosis from sustained calcium imbalance and oxidative damage, with elevated risk in calcium-sensitive tissues like the heart and brain due to heightened vulnerability to ER-mediated apoptosis.

Mitigation Strategies

To mitigate the systemic of thapsigargin, which arises from its potent and non-selective inhibition of pumps leading to widespread disruption of calcium , researchers have developed strategies that restrict activation to tumor microenvironments. These prodrugs conjugate thapsigargin to or chemical masks that are cleaved by cancer-specific enzymes, thereby minimizing exposure to healthy tissues while preserving cytotoxic effects against malignant cells. A prominent example is mipsagargin (G202), a activated by prostate-specific membrane antigen (PSMA), which is overexpressed in and certain other solid tumors. Upon intravenous administration, mipsagargin remains inactive until PSMA-mediated releases the active thapsigargin derivative (8-O-debenzoylthapsigargin) at the tumor site, achieving tumor growth inhibition in preclinical xenograft models without significant off-target toxicity. Phase I and II clinical trials (completed by 2018; e.g., NCT02067156) in patients with advanced and other PSMA-expressing cancers demonstrated acceptable tolerability, with dose-limiting toxicities primarily limited to mild infusion reactions and manageable liver enzyme elevations, though no further phases advanced as of 2025. Another approach involves (PSA)-activated prodrugs, such as those linking thapsigargin to peptides cleaved by PSA, a elevated in . Preclinical studies with PSA-prodrugs like Leu-12ADT showed complete regression of prostate tumor xenografts in mice at doses fourfold higher than the maximum tolerated dose of unmodified thapsigargin, with no observed systemic due to selective activation in PSA-rich environments. These strategies highlight the potential of enzyme-triggered release to enhance the of thapsigargin, though challenges remain in optimizing linker stability and enzyme specificity for broader applications. As of 2025, mipsagargin has not received regulatory approval, with development appearing discontinued following completion of Phase II trials showing limited efficacy despite tolerability. In laboratory and handling contexts, thapsigargin's —manifesting as , respiratory distress, and potential carcinogenicity—necessitates strict precautions to prevent inadvertent exposure. Standard protocols recommend using it in well-ventilated fume hoods, wearing gloves, protective eyewear, and lab coats, followed by thorough handwashing and of surfaces with appropriate solvents like DMSO or . Storage in desiccated, amber vials at -20°C further stabilizes the compound and reduces degradation-related risks during research use.

References

  1. https://pmc.ncbi.nlm.nih.gov/articles/PMC7792614/
  2. https://www.sciencedirect.com/topics/chemistry/thapsigargin
  3. https://pmc.ncbi.nlm.nih.gov/articles/PMC7913267/
  4. https://www.mdpi.com/2072-6694/11/6/833
  5. https://ctv.veeva.com/study/efficacy-safety-and-cns-exposure-of-g-202-mipsagargin-in-patients-with-recurrent-or-progressive-g
  6. https://veterinaryresearch.biomedcentral.com/articles/10.1186/s13567-024-01359-x
  7. https://pubs.acs.org/doi/10.1021/acscentsci.6b00313
  8. https://pubmed.ncbi.nlm.nih.gov/79299/
  9. https://pubmed.ncbi.nlm.nih.gov/26429715/
  10. https://pmc.ncbi.nlm.nih.gov/articles/PMC6272310/
  11. https://www.cheminst.ca/magazine/article/the-life-saving-potential-of-the-deadly-carrot/
  12. https://www.semanticscholar.org/paper/Thapsigargin%252C-Origin%252C-Chemistry%252C-Structure-Activity-Doan-Christensen/a6b6f79c43339b030d0af1a2402c7b727ac83a65
  13. https://pubmed.ncbi.nlm.nih.gov/11474736/
  14. https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/thapsia-garganica
  15. https://pubchem.ncbi.nlm.nih.gov/compound/Thapsigargin
  16. https://www.pnas.org/doi/10.1073/pnas.0403300101
  17. https://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5351
  18. https://www.researchgate.net/publication/282568293_Thapsigargin_Origin_Chemistry_Structure-Activity_Relationships_and_Prodrug_Development
  19. https://www.sigmaaldrich.com/US/en/product/sigma/t9033
  20. https://www.cellsignal.com/products/12758/datasheet
  21. https://www.selleckchem.com/products/thapsigargin.html
  22. https://www.enzo.com/product/thapsigargin/
  23. https://www.sciencedirect.com/topics/neuroscience/thapsigargin
  24. https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/242/310/t9033pis.pdf
  25. https://www.mdpi.com/1422-0067/22/1/4
  26. https://www.cretanflora.com/thapsia_garganica.html
  27. https://pfaf.org/user/Plant.aspx?LatinName=Thapsia%20garganica
  28. https://www.researchgate.net/publication/275240902_Thapsigargin-From_Thapsia_L_to_Mipsagargin
  29. https://portals.iucn.org/library/efiles/documents/2011-014.pdf
  30. https://www.mdpi.com/1420-3049/20/4/6113
  31. https://bio-protocol.org/exchange/minidetail?id=4178832&type=30
  32. https://www.sciencedirect.com/science/article/abs/pii/S1570023213001116
  33. https://plantmethods.biomedcentral.com/articles/10.1186/s13007-018-0346-z
  34. https://pmc.ncbi.nlm.nih.gov/articles/PMC6128993/
  35. https://doi.org/10.3390/molecules24213961
  36. https://doi.org/10.1042/BJ20120654
  37. https://doi.org/10.3390/molecules20046113
  38. https://doi.org/10.1007/s00425-016-2651-1
  39. https://jhoonline.biomedcentral.com/articles/10.1186/s13045-020-01015-9
  40. https://pmc.ncbi.nlm.nih.gov/articles/PMC5712609/
  41. https://pubmed.ncbi.nlm.nih.gov/8405462/
  42. https://pmc.ncbi.nlm.nih.gov/articles/PMC5409463/
  43. https://www.researchgate.net/publication/21322651_Thapsigargin_inhibits_the_sarcoplasmic_or_endoplasmic_reticulum_Ca-ATPase_family_of_calcium_pumps
  44. https://pubmed.ncbi.nlm.nih.gov/28264934/
  45. https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2018.00669/full
  46. https://pubmed.ncbi.nlm.nih.gov/9935265/
  47. https://pmc.ncbi.nlm.nih.gov/articles/PMC3124681/
  48. https://pubmed.ncbi.nlm.nih.gov/28302665/
  49. https://journals.biologists.com/jcs/article/130/7/1209/56663/New-insights-into-autophagosome-lysosome-fusion
  50. https://pubmed.ncbi.nlm.nih.gov/8625480/
  51. https://link.springer.com/article/10.1007/s12032-024-02541-z
  52. https://link.springer.com/article/10.1007/s00262-021-03072-6
  53. https://www.nature.com/articles/bjc201672
  54. https://clinicaltrials.gov/study/NCT02067156
  55. https://www.mdpi.com/1420-3049/26/24/7469
  56. https://www.researchgate.net/publication/309093697_Thapsigargin_sensitizes_human_esophageal_cancer_to_TRAIL-induced_apoptosis_via_AMPK_activation
  57. https://pmc.ncbi.nlm.nih.gov/articles/PMC6874456/
  58. https://www.nottingham.ac.uk/news/thapsigargin-covid-19
  59. https://www.researchgate.net/publication/23393281_Methods_for_Studying_Store-Operated_Calcium_Entry
  60. https://www.organic-chemistry.org/totalsynthesis/totsyn03/thapsigargin-ley.shtm
  61. https://pubs.acs.org/doi/10.1021/jacs.7b01734
  62. https://www.researchgate.net/publication/342115771_Thapsigargins_and_induced_chemical_defence_in_Thapsia_garganica
  63. https://www.fermentek.com/sites/default/files/2024-08/SDS%20Thapsigargin%20vers%208-2024.pdf
  64. https://pmc.ncbi.nlm.nih.gov/articles/PMC6986015/
  65. https://www.sciencedirect.com/science/article/abs/pii/S0009279724004381
  66. https://www.nature.com/articles/s41420-025-02602-w
  67. https://pmc.ncbi.nlm.nih.gov/articles/PMC9500646/
  68. https://pmc.ncbi.nlm.nih.gov/articles/PMC3276380/
  69. https://academic.oup.com/jnci/article/95/13/990/2520317
  70. https://www.sigmaaldrich.com/BE/en/sds/sigma/t9033
  71. https://www.fishersci.com/store/msds?partNumber=BP6291&productDescription=THAPSIGARGIN%2B1MG&vendorId=VN00033897&countryCode=US&language=en
Add your contribution
Related Hubs
User Avatar
No comments yet.