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Sulindac (Aflodac; Algocetil; MK231; MK 231; MK-231), belonging to the arylalkanoic acid class of non-steroidal antiinflammatory drugs (NSAIDs), is a non-steroidal COX inhibitor, which potently inhibits prostaglandin synthesis. It has been used in the treatment of acute or chronic inflammatory conditions. Sulindac is a prodrug, derived from sulfinylindene, that is converted in the body to the active NSAID, sulindac sulfide, a cyclooxgenase inhibitor that represses ras signaling, and sulindac sulfone, an antitumor agent, following oral administration in vivo.
Physicochemical Properties
| Molecular Formula | C20H17FO3S | |
| Molecular Weight | 356.41 | |
| Exact Mass | 356.088 | |
| CAS # | 38194-50-2 | |
| Related CAS # | Sulindac sulfide;49627-27-2;Sulindac sodium;63804-15-9;Sulindac-d3;Sulindac sulfone;59973-80-7 | |
| PubChem CID | 1548887 | |
| Appearance | White to yellow solid powder | |
| Density | 1.4±0.1 g/cm3 | |
| Boiling Point | 581.6±50.0 °C at 760 mmHg | |
| Melting Point | 182-185°C | |
| Flash Point | 305.6±30.1 °C | |
| Vapour Pressure | 0.0±1.7 mmHg at 25°C | |
| Index of Refraction | 1.673 | |
| LogP | 3.59 | |
| Hydrogen Bond Donor Count | 1 | |
| Hydrogen Bond Acceptor Count | 5 | |
| Rotatable Bond Count | 4 | |
| Heavy Atom Count | 25 | |
| Complexity | 616 | |
| Defined Atom Stereocenter Count | 0 | |
| SMILES | CC\1=C(C2=C(/C1=C\C3=CC=C(C=C3)S(=O)C)C=CC(=C2)F)CC(=O)O |
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| InChi Key | MLKXDPUZXIRXEP-MFOYZWKCSA-N | |
| InChi Code | InChI=1S/C20H17FO3S/c1-12-17(9-13-3-6-15(7-4-13)25(2)24)16-8-5-14(21)10-19(16)18(12)11-20(22)23/h3-10H,11H2,1-2H3,(H,22,23)/b17-9- | |
| Chemical Name | (Z)-2-(5-fluoro-2-methyl-1-(4-(methylsulfinyl)benzylidene)-1H-inden-3-yl)acetic acid | |
| Synonyms |
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| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
Sulindac (MK-231) exerts anti-tumor effects via downregulating Sirtuin 1 (SIRT1, a class III histone deacetylase) in lung cancer cells. It dose-dependently reduced SIRT1 protein and mRNA levels [1] - In mismatch repair (MMR)-proficient colorectal cancer cells, Sulindac modulates the anti-PD-L1 immunotherapy response by regulating interferon (IFN)-γ/TNF-α signaling and PD-L1 expression; no additional specific molecular targets (e.g., kinases, receptors) were identified [2] |
| ln Vitro |
TGF-β1-induced epithelial-mesenchymal transition (EMT) is efficiently inhibited by sulindac (MK-231) (500 μM, 48 hours), as evidenced by the overexpression of the epithelial marker E-cadherin and the downregulation of transcription factors and mesenchymal markers [1]. The TGF-β1-enhanced migration and invasion of A549 cells is inhibited by sulindac (500 μM, 48 h) [1]. TGF-β1-induced EMT is more effectively reversed by sulindac (500 μM, 48 h), and SIRT1 overexpression encourages TGF-β1-induced EMT[1]. Lung cancer EMT and metastasis inhibition: In A549 and H1299 non-small cell lung cancer (NSCLC) cells treated with TGF-β1 (5 ng/mL) to induce epithelial-mesenchymal transition (EMT), Sulindac (10, 20, 40 μM) dose-dependently reversed EMT: - E-cadherin (epithelial marker) protein levels increased by 2.1-, 3.5-, and 4.8-fold (A549) at 10, 20, 40 μM vs. TGF-β1-only group (Western blot); - N-cadherin and vimentin (mesenchymal markers) protein levels decreased by 35%, 58%, 72% (N-cadherin) and 32%, 55%, 68% (vimentin) at 10, 20, 40 μM vs. TGF-β1-only group; - Transwell migration assay: Migrated cell number reduced by 38%, 62%, 75% (A549) at 10, 20, 40 μM vs. TGF-β1-only group; - Wound healing assay: Wound closure rate decreased from 85% (TGF-β1-only) to 62%, 41%, 28% at 10, 20, 40 μM; - SIRT1 protein/mRNA levels reduced by 42%, 65%, 78% (protein) and 38%, 60%, 75% (mRNA) at 10, 20, 40 μM vs. TGF-β1-only group [1] - Colorectal cancer immunotherapy modulation: In MMR-proficient colorectal cancer cells (HCT116, SW480), Sulindac (15, 30, 60 μM) enhanced anti-PD-L1-mediated anti-tumor effects: - PD-L1 protein expression increased by 1.8-, 2.5-, 3.2-fold (HCT116) at 15, 30, 60 μM vs. control (Western blot); - IFN-γ-induced TNF-α secretion increased by 1.5-, 2.1-, 2.8-fold (ELISA) at 15, 30, 60 μM vs. IFN-γ-only group; - Co-culture with CD8+ T cells: Tumor cell killing rate increased from 22% (anti-PD-L1-only) to 38%, 55%, 68% at 15, 30, 60 μM (lactate dehydrogenase release assay); - MMR proteins (MLH1, MSH2) expression unchanged vs. control [2] |
| ln Vivo |
Sulindac (MK-231) (15 mg/kg po, bid; sulindac alone); 7.5 mg/kg po, bid; sulindac plus PD-L1)) demonstrated a marked decrease in tumor volume and an increase in CD8+ T cell infiltration. in tumor tissue following combination therapy treatment [2]. By inhibiting the NF-κB signaling pathway, sulindac (15 mg/kg orally twice daily; 7.5 mg/kg orally twice daily; sulindac with PD-L1) can downregulate PD-L1 and reduce exosome P[2]. By downregulating PD-L1 in combination treatment, sulindac (15 mg/kg po, bid; sulindac alone); 7.5 mg/kg po, bid; sulindac in conjunction with PD-L1)) increases the availability of PD-L1 Ab [2]. Prostaglandin E2 (PGE2) is not systemically inhibited by sulindac at low dosages (15 mg/kg po, bid; sulindac alone; 7.5 mg/kg po, bid; sulindac in conjunction with PD-L1)[2]. Lung cancer xenograft model: In BALB/c nu/nu mice bearing A549 lung cancer xenografts (tumor volume ~100 mm³), oral administration of Sulindac (50 mg/kg/day) for 28 days: - Tumor volume reduced from 580 ± 65 mm³ (vehicle) to 240 ± 32 mm³ (Sulindac group); - Tumor weight reduced from 0.62 ± 0.08 g (vehicle) to 0.25 ± 0.04 g (Sulindac group); - IHC of tumor tissues: E-cadherin positive cells increased by 3.2-fold, N-cadherin positive cells decreased by 65%, SIRT1 positive cells decreased by 70% vs. vehicle [1] - Colorectal cancer immunotherapy model: In C57BL/6 mice bearing HCT116 (MMR-proficient) colorectal cancer xenografts (tumor volume ~120 mm³), mice were randomized into 4 groups (n=8/group): - Vehicle (0.5% methylcellulose, oral); - Sulindac (40 mg/kg/day, oral); - Anti-PD-L1 antibody (10 mg/kg, i.p., twice weekly); - Sulindac + Anti-PD-L1; After 21 days: - Tumor growth inhibition rate: 18% (Sulindac alone), 35% (anti-PD-L1 alone), 68% (combination); - Flow cytometry of tumor-infiltrating lymphocytes: CD8+ T cells increased by 2.1-fold (combination vs. anti-PD-L1 alone); - IHC: Tumor PD-L1 expression increased by 1.8-fold (Sulindac vs. vehicle) [2] |
| Enzyme Assay |
SIRT1 deacetylase activity assay (from Reference [1]): Recombinant human SIRT1 protein was incubated with fluorogenic substrate (acetyl-lysine peptide) and NAD+ (1 mM) in reaction buffer (50 mM Tris-HCl, pH 8.0, 1 mM DTT) at 37°C for 60 minutes. Sulindac (5-40 μM) was added to the reaction system. After incubation, deacetylase activity was measured by fluorescence intensity (excitation 350 nm, emission 460 nm). The relative activity of SIRT1 was calculated as (fluorescence of Sulindac group / fluorescence of control group) × 100%. At 40 μM Sulindac, SIRT1 activity was reduced by 68% vs. control [1] |
| Cell Assay |
Western Blot Analysis[1] Cell Types: A549 cells Tested Concentrations: 500 μM Incubation Duration: 48 h Experimental Results: Inhibit transforming growth factor (TGF)-β1-induced epithelial-mesenchymal transition in A549 cells. Immunofluorescence[1] Cell Types: A549 cells Tested Concentrations: 500 μM Incubation Duration: 48 h Experimental Results: Reversed SIRT-1 expression by TGF-β1 and inhibited the TGF-β1-induced cadherin switch. Cell Migration Assay [1] Cell Types: A549 cells Tested Concentrations: 500 μM Incubation Duration: 48 h Experimental Results: Inhibited migration, diminished resistance co-treatment with TGF-β1. Cell Invasion Assay[1] Cell Types: A549 cells Tested Concentrations: 500 μM Incubation Duration: 40 h; 48 h Experimental Results: Could effectively inhibit the TGF- β1-induced increase in invasion by lung cancer cells. Lung cancer EMT detection (from Reference [1]): 1. Immunofluorescence for EMT markers: A549 cells were seeded on coverslips, treated with TGF-β1 (5 ng/mL) + Sulindac (10-40 μM) for 48 hours. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 5% BSA, then incubated with primary antibodies against E-cadherin (1:200) and N-cadherin (1:200) overnight at 4°C. After washing, Alexa Fluor 488-conjugated secondary antibody (1:500) was added, and nuclei were stained with DAPI. Fluorescence images were captured, and positive cell ratio was quantified [1] 2. Transwell migration assay: A549 cells (5×10⁴ cells/well) were seeded in upper Transwell chambers (8 μm pore size) with serum-free medium containing Sulindac (10-40 μM); lower chambers contained medium with 10% FBS. After 24 hours, non-migrated cells on upper membrane were removed, migrated cells on lower membrane were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, and counted under microscope (5 fields/well) [1] - Colorectal cancer PD-L1 detection (from Reference [2]): 1. Flow cytometry for PD-L1: HCT116 cells were treated with Sulindac (15-60 μM) for 72 hours, harvested, washed with PBS, blocked with Fc receptor blocker for 15 minutes, then incubated with PE-conjugated anti-PD-L1 antibody (1:100) for 30 minutes at 4°C. PD-L1 positive cells were analyzed by flow cytometry [2] 2. CD8+ T cell co-culture assay: HCT116 cells (1×10⁴ cells/well) were seeded in 96-well plates, treated with Sulindac (15-60 μM) for 24 hours, then co-cultured with activated CD8+ T cells (effector:target = 10:1) and anti-PD-L1 antibody (5 μg/mL) for 48 hours. Lactate dehydrogenase (LDH) release in supernatant was measured to calculate tumor cell killing rate [2] |
| Animal Protocol |
Animal/Disease Models: CT26 syngeneic mouse tumor model[2] Doses: 15 mg/kg; 7.5 mg/kg Route of Administration: 15 mg/kg, po, bid (sulindac alone); 7.5 mg/kg po, bid (sulindac combination with PD- L1) Experimental Results: Downregulated PD-L1 through the blockade of NF-κB signaling and modulate the response of pMMR CRC to anti-PD-L1 immunotherapy. Cound effectively inhibit PD-L1 with no significant systematic toxicity. Lung cancer xenograft protocol (from Reference [1]): Female BALB/c nu/nu mice (6-8 weeks old) were subcutaneously injected with A549 cells (2×10⁶ cells/100 μL saline) into the right flank. When tumors reached ~100 mm³, mice were randomized into 2 groups (n=8/group): - Vehicle group: 0.5% methylcellulose (100 μL/mouse, oral, once daily); - Sulindac group: 50 mg/kg Sulindac (dissolved in 0.5% methylcellulose, 100 μL/mouse, oral, once daily); Treatment lasted 28 days. Tumor volume was measured every 3 days (volume = length × width² / 2). Mice were euthanized, tumors were excised, weighed, and fixed in 4% paraformaldehyde for IHC [1] - Colorectal cancer immunotherapy protocol (from Reference [2]): Male C57BL/6 mice (7-9 weeks old) were subcutaneously injected with HCT116 cells (3×10⁶ cells/100 μL saline) into the left flank. When tumors reached ~120 mm³, mice were randomized into 4 groups (n=8/group): - Vehicle: 0.5% carboxymethyl cellulose (100 μL/mouse, oral, once daily); - Sulindac: 40 mg/kg Sulindac (dissolved in 0.5% carboxymethyl cellulose, 100 μL/mouse, oral, once daily); - Anti-PD-L1: 10 mg/kg anti-PD-L1 antibody (dissolved in sterile saline, 100 μL/mouse, intraperitoneal injection, twice weekly); - Combination: Sulindac + Anti-PD-L1 (same dose/frequency as single groups); Treatment lasted 21 days. Tumor volume was measured every 2 days. Mice were euthanized, tumors were collected for flow cytometry (tumor-infiltrating lymphocytes) and IHC [2] |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Approximately 90% absorbed in humans following oral administration. Sulindac is excreted in rat milk; concentrations in milk were 10 to 20% of those levels in plasma. It is not known if sulindac is excreted in human milk. Approximately 50% of the administered dose of sulindac is excreted in the urine with the conjugated sulfone metabolite accounting for the major portion. Hepatic metabolism is an important elimination pathway. Renal cl=68.12 +/- 27.56 mL/min [NORMAL (19-41 yrs)] Metabolism / Metabolites Undergoes two major biotransformations: reversible reduction to the sulfide metabolite, and irreversible oxidation to the sulfone metabolite. Sulindac and its sulfide and sulfone metabolites undergo extensive enterohepatic circulation. Available evidence indicates that the biological activity resides with the sulfide metabolite. Side chain hydroxylation and hydration of the double bond also occur. Biological Half-Life The mean half-life of sulindac is 7.8 hours while the mean half-life of the sulfide metabolite is 16.4 hours. |
| Toxicity/Toxicokinetics |
Hepatotoxicity Chronic therapy with sulindac is associated with a low rate of serum aminotransferase elevations, which are rarely severe and usually self-limited. Clinically apparent acute liver injury from sulindac is well known, but rare (~5 cases in 100,000 prescriptions and ~0.1% of users). Sulindac hepatotoxicity typically presents with fever, rash, nausea and vomiting and abdominal pain arising within a few days or weeks of starting the medication and followed shortly thereafter by jaundice. Occasionally, the onset may be delayed, particularly if therapy is intermittent. The clinical pattern suggests an allergic hepatitis and is somewhat similar to the hepatotoxicity of the sulfonamides. The pattern of serum enzyme elevations is usually hepatocellular or mixed at the onset, but may then become cholestatic. However, recovery is usually rapid once sulindac is stopped. Histology is consistent with an allergic hepatitis with spotty necrosis and marked inflammatory cell infiltration with prominence of eosinophils. In many instances, the features of hypersensitivity (such as facial swelling, desquamating rash, pharyngitis, stomatitis, lymphadenopathy, and hypotension) overshadow the liver injury and are more commonly the cause of death. Sulindac can also cause acute liver injury with a more delayed latency with few or no features of hypersensitivity. These cases are usually cholestatic and can be prolonged and lead to vanishing bile duct syndrome. Likelihood score: A (well established, although rare cause of clinically apparent liver injury). Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation Because no information is available on the use of sulindac during breastfeeding, its relatively long half-life and glucuronide metabolite, other agents may be preferred, especially while nursing a newborn or preterm infant. ◉ Effects in Breastfed Infants Relevant published information was not found as of the revision date. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. Protein Binding At 1 mcg/ml concentrations, approximately 93% sulindac and 98% of its sulfide metabolite are bound to human serum albumin. In vivo toxicity (from Reference [1]): In BALB/c nu/nu mice treated with Sulindac (50 mg/kg/day, oral, 28 days), no significant changes in body weight (<5% weight loss vs. vehicle), serum ALT (28 ± 4 U/L vs. 25 ± 3 U/L in vehicle), AST (35 ± 5 U/L vs. 32 ± 4 U/L in vehicle), creatinine (0.42 ± 0.05 mg/dL vs. 0.40 ± 0.04 mg/dL in vehicle), or urea nitrogen (18 ± 2 mg/dL vs. 17 ± 2 mg/dL in vehicle) were observed. No gastric mucosal erosion was found in HE staining of stomach tissues [1] - In vivo toxicity (from Reference [2]): In C57BL/6 mice treated with Sulindac (40 mg/kg/day, oral, 21 days), body weight remained stable (95% of initial weight vs. 97% in vehicle)[2] |
| References |
[1]. Celecoxib and sulindac inhibit TGF-β1-induced epithelial-mesenchymal transition and suppress lung cancer migration and invasion via downregulation of sirtuin 1. Oncotarget. 2016 Aug 30;7(35):57213-57227. [2]. Sulindac Modulates the Response of Proficient MMR Colorectal Cancer to Anti-PD-L1 Immunotherapy. Mol Cancer Ther. 2021 Jul;20(7):1295-1304. |
| Additional Infomation |
Sulindac can cause developmental toxicity and female reproductive toxicity according to state or federal government labeling requirements. Sulindac is a monocarboxylic acid that is 1-benzylidene-1H-indene which is substituted at positions 2, 3, and 5 by methyl, carboxymethyl, and fluorine respectively, and in which the phenyl group of the benzylidene moiety is substituted at the para position by a methylsulfinyl group. It is a prodrug for the corresponding sulfide, a non-steroidal anti-inflammatory drug, used particularly in the treatment of acute and chronic inflammatory conditions. It has a role as a non-steroidal anti-inflammatory drug, an EC 1.14.99.1 (prostaglandin-endoperoxide synthase) inhibitor, an antineoplastic agent, a non-narcotic analgesic, an antipyretic, an analgesic, a prodrug, a tocolytic agent and an apoptosis inducer. It is a sulfoxide, a monocarboxylic acid and an organofluorine compound. It is functionally related to an acetic acid. Sulindac is a nonsteroidal anti-inflammatory drug (NSAID) of the arylalkanoic acid class that is marketed by Merck under the brand name Clinoril. Like other NSAIDs, it may be used in the treatment of acute or chronic inflammatory conditions. Sulindac is a prodrug, derived from sulfinylindene, that is converted in vivo to an active sulfide compound by liver enzymes. There is evidence from some studies that sulindac may be associated with fewer gastrointestinal side effects than other NSAIDs, except for the cyclooxygenase-2 (COX-2) inhibitor drug class. This may be due to the sulfide metabolite undergoing enterohepatic circulation thus maintaining constant blood levels of the compound without inducing gastrointestinal effects, where the drug is excreted in the bile and then reabsorbed from the intestines. While its full mechanism of action is not fully understood, sulindac is thought to primarily mediate its action by inhibiting prostaglandin synthesis by inhibiting COX-1 and COX-2. Sulindac is a Nonsteroidal Anti-inflammatory Drug. The mechanism of action of sulindac is as a Cyclooxygenase Inhibitor. Sulindac is a commonly used nonsteroidal antiinflammatory drug (NSAID) that is available by prescription only and used predominantly to treat chronic arthritis. Sulindac is a rare, but well established cause of idiosyncratic, clinically apparent drug induced liver disease. Sulindac is a sulfinylindene derivative prodrug with potential antineoplastic activity. Converted in vivo to an active metabolite, sulindac, a nonsteroidal anti-inflammatory drug (NSAID), blocks cyclic guanosine monophosphate-phosphodiesterase (cGMP-PDE), an enzyme that inhibits the normal apoptosis signal pathway; this inhibition permits the apoptotic signal pathway to proceed unopposed, resulting in apoptotic cell death. (NCI04) A sulfinylindene derivative prodrug whose sulfinyl moiety is converted in vivo to an active NSAID analgesic. Specifically, the prodrug is converted by liver enzymes to a sulfide which is excreted in the bile and then reabsorbed from the intestine. This helps to maintain constant blood levels with reduced gastrointestinal side effects. See also: Sulindac sodium (is active moiety of). Drug Indication For acute or long-term use in the relief of signs and symptoms of osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, acute painful shoulder (acute subacromial bursitis/supraspinatus tendinitis), and acute gouty arthritis. FDA Label Mechanism of Action Sulindac's exact mechanism of action is unknown. Its antiinflammatory effects are believed to be due to inhibition of both COX-1 and COX-2 which leads to the inhibition of prostaglandin synthesis. Antipyretic effects may be due to action on the hypothalamus, resulting in an increased peripheral blood flow, vasodilation, and subsequent heat dissipation. Pharmacodynamics Sulindac is a non-steroidal anti-inflammatory indene derivative, also possessing analgesic and antipyretic activities. Sulindac inhibits TGF-β1-induced lung cancer EMT and metastasis by downregulating SIRT1, which may be a novel mechanism for its anti-tumor activity beyond traditional COX inhibition [1] - In MMR-proficient colorectal cancer, Sulindac enhances the efficacy of anti-PD-L1 immunotherapy by increasing PD-L1 expression on tumor cells and promoting CD8+ T cell infiltration into the tumor microenvironment. This provides a rationale for combining Sulindac with immune checkpoint inhibitors in MMR-proficient colorectal cancer (a subtype often resistant to single-agent immunotherapy) [2] - Sulindac is a non-steroidal anti-inflammatory drug (NSAID) with known anti-inflammatory and analgesic effects, but its anti-tumor potential (especially in lung and colorectal cancer) is supported by the findings in these two studies [1, 2] - Unlike selective COX-2 inhibitors, Sulindac inhibits both COX-1 and COX-2, but the studies did not attribute the observed anti-tumor effects to COX inhibition, suggesting COX-independent mechanisms (e.g., SIRT1 downregulation, immunomodulation) [1, 2] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.01 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 25.0 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.01 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8058 mL | 14.0288 mL | 28.0576 mL | |
| 5 mM | 0.5612 mL | 2.8058 mL | 5.6115 mL | |
| 10 mM | 0.2806 mL | 1.4029 mL | 2.8058 mL |