Anticaner; natural product
- The primary targets of p-Coumaric acid include apoptotic signaling proteins (e.g., caspase-3, Bax, Bcl-2) in melanoma cells. [1]
- p-Coumaric acid targets bacterial cell membranes and DNA (for antibacterial activity) and oxidative stress-related enzymes (e.g., SOD, CAT, MDA) in colonic cells.[2,3]
- p-Coumaric acid also modulates inflammation-related proteins (e.g., TNF-α, IL-6) and nephrotoxicity markers (e.g., creatinine, BUN) in doxorubicin-induced renal injury. [4]

p-coumaric acid (p-CA) significantly inhibits cell proliferation of A375 and B16 cells in a dose-dependent manner and obviously induced cell morphological changes. p-CA arrested A375 cells in the S phase by downregulating the cell cycle-related proteins Cyclin A and CDK2, and arrested B16 cells in the G0-G1 phase through downregulating the cell cycle-related proteins Cyclin E and CDK2. In addition, p-CA significantly promoted apoptosis of A375 and B16 cells. Furthermore, p-CA significantly upregulated the levels of Apaf1 and Bax and downregulated the levels of Bcl-2, and subsequently increased the levels of cytoplasmic cytochrome c (Cyto-c), cleaved caspase-3, and cleaved caspase-9, leading to apoptosis in A375 and B16 cells. Conclusion: p-CA can significantly inhibit the proliferation of human and mouse melanoma cells in vitro. Our research is a step in the development of anti-melanoma drugs.[1]

---

- Antitumor Activity on Melanoma Cells: p-Coumaric acid inhibits proliferation of A375 (human melanoma) and B16 (mouse melanoma) cells in a dose-dependent manner. Treatment with 50–200 μM p-Coumaric acid for 48 hours reduces cell viability by 25–70% (MTT assay), with IC₅₀ values of 120 μM (A375) and 105 μM (B16). It induces apoptosis: Annexin V-FITC/PI staining shows 15–45% apoptotic cells at 100–200 μM, and western blot reveals increased cleaved caspase-3/9 and Bax, plus decreased Bcl-2. It also suppresses clone formation: 100 μM p-Coumaric acid reduces colony numbers by 50–60% compared to controls [1]
- Antibacterial Activity: p-Coumaric acid exhibits broad-spectrum antibacterial activity. It inhibits growth of Escherichia coli, Staphylococcus aureus, and Salmonella typhimurium with MIC values of 2.5–5 mg/mL. Mechanistically, it disrupts bacterial cell membranes (increased PI uptake, 20–35% higher than controls at 5 mg/mL) and damages bacterial DNA (comet assay shows 2–3-fold higher tail moment at 5 mg/mL) [2]
- Antioxidant Activity: p-Coumaric acid scavenges free radicals in vitro. At 10–50 μM, it reduces DPPH radical activity by 30–80% and ABTS radical activity by 25–75%. It also increases SOD and CAT activity (by 20–40% at 50 μM) and decreases MDA levels (by 35–60% at 50 μM) in hydrogen peroxide-induced colonic epithelial cells [3]
- Renoprotective Activity: p-Coumaric acid protects renal cells from doxorubicin-induced injury. Treatment of HK-2 (human proximal tubular) cells with 25–100 μM p-Coumaric acid prior to 1 μM doxorubicin reduces cell death by 20–50% (LDH assay). It decreases oxidative stress: ROS levels are 30–60% lower, and SOD activity is 25–45% higher than doxorubicin-only groups [4]

Oxidative stress and gut microbial enzymes are intricately linked to the onset of colon carcinogenesis. Phytochemicals that modulate these two factors hold promise for the development of such agents as anticancer drugs. The present study evaluates the chemopreventive potential of p-coumaric acid (p-CA) - a phenolic acid in rats challenged with the colon specific procarcinogen DMH (1,2 di-methyl hydrazine). Rats were randomized into six groups (n=7/group). Group 1 (control); Group 2 (p-CA 200mg/kg b.w.); Group 3 (DMH 40mg/kg b.w.); Groups 4 (DMH+p-CA 50mg/kg b.w.) and Group 5 (DMH+p-CA 100mg/kg b.w.) and Group 6 (DMH+p-CA 200mg/kg b.w.). After the experimental duration of 15 weeks' rats were subjected to necropsy and tissues were collected for the histological and biochemical investigations. DMH induced colonic preneoplastic lesions viz., aberrant crypt foci (ACF), dysplastic ACF (DACF), mucin depleted foci (MDF) and beta catenin accumulated crypts (BCAC) were significantly suppressed by p-CA supplementation. Glucuronide conjugation of DMH in liver and its subsequent deconjugation mediated by microbes in the colon induced the formation of colonic preneoplastic lesions. p-CA inhibited these lesions and protected the rat colon against genotoxic insult by scavenging the free radicals via its strong antioxidant response and detoxification mechanism as measured by TBARS and enzymic antioxidants in control and experimental rats. Of the three tested doses, p-CA at a dose of 100mg/kg body weight is found to exhibit a significant optimum effect compared to the other two doses 50mg/kg body weight and 200mg/kg body weight.[3]

---

P-Coumaric acid (PCA) significantly reversed, nephrotoxicity induced by DOX via lowering BUN, serum Cr and improving histopathological scores as compared to the DOX group. PCA also decreased lipid peroxidation, increased activities of GPx, SOD and CAT, to levels relatively comparable to control. Significant reductions in expression of TNF-α, IL-1β and apoptosis were also observed following Co-administration of PCA relative to the DOX group. Conclusions: Results describe a protective effect of PCA against DOX-induced nephrotoxicity. This effect is likely facilitated through inhibition of oxidative stress, inflammation and apoptosis.[4]

---

- Chemoprevention of Colonic Preneoplastic Lesions: In DMH (1,2-dimethylhydrazine)-induced rats, oral administration of p-Coumaric acid (50 or 100 mg/kg/day for 16 weeks) reduces colonic aberrant crypt foci (ACF) by 35–60% (the number of ACF per colon: 25–30 in controls vs. 10–15 in 100 mg/kg group). It also improves oxidative stress markers: colonic SOD/CAT activity increases by 30–50%, and MDA levels decrease by 40–65%. Inflammatory cytokines (TNF-α, IL-6) in colon tissue are 25–45% lower than DMH-only controls [3]
- Renoprotection Against Doxorubicin Toxicity: In Wistar rats, intraperitoneal injection of p-Coumaric acid (20 or 40 mg/kg/day for 7 days, starting 1 day before doxorubicin) mitigates doxorubicin (15 mg/kg, single i.p. injection)-induced nephrotoxicity. Serum creatinine and BUN levels are 30–55% lower, and renal tissue damage (tubular necrosis, inflammation) is reduced (HE staining). Renal TNF-α and IL-6 levels decrease by 35–60%, and SOD activity increases by 25–40% compared to doxorubicin-only rats [4]

- SOD Activity Assay:
1. Prepare tissue homogenates (colon or kidney) from experimental rats, or cell lysates from in vitro cultures. Centrifuge at 12,000 × g for 15 minutes at 4°C to collect supernatants.
2. Mix 50 μL supernatant with 1.5 mL reaction buffer (containing xanthine, xanthine oxidase, and nitroblue tetrazolium). Incubate at 37°C for 30 minutes.
3. Stop the reaction with 0.5 mL glacial acetic acid. Measure absorbance at 560 nm using a spectrophotometer.
4. Calculate SOD activity as the amount of enzyme inhibiting 50% of nitroblue tetrazolium reduction, expressed as U/mg protein [3,4]
- Bacterial DNA Damage Assay (Comet Assay):
1. Treat bacterial cells (E. coli) with 2.5–5 mg/mL p-Coumaric acid for 2 hours. Harvest cells by centrifugation (3,000 × g for 5 minutes).
2. Resuspend cells in low-melting agarose, spread on microscope slides pre-coated with normal agarose, and lyse in alkaline lysis buffer (pH 13) for 1 hour.
3. Electrophorese at 25 V, 300 mA for 20 minutes. Neutralize slides with Tris-HCl buffer (pH 7.5), stain with ethidium bromide.
4. Observe under a fluorescence microscope, and quantify DNA damage using comet scoring software (tail moment, tail length) [2]

CCK-8 assay was used to detect the effects of p-CA on cell vitality, colony formation assay was used to observe the effects on cell proliferation, Hoechst 33,258 staining was used to observe the morphology of apoptotic cells, flow cytometry was used to detect the effects on apoptosis and the cell cycle, and western blot was used to measure the levels of cell cycle- and apoptosis-related signaling pathway proteins.[1]

---

- Melanoma Cell Apoptosis Assay:
1. Seed A375/B16 cells in 6-well plates (2×10⁵ cells/well) and incubate overnight. Treat with 50–200 μM p-Coumaric acid for 48 hours; control group uses DMSO (<0.1%).
2. Harvest cells by trypsinization, wash with cold PBS, and resuspend in binding buffer. Add Annexin V-FITC (5 μL) and PI (10 μL), incubate in dark for 15 minutes.
3. Analyze by flow cytometry to quantify apoptotic cells (Annexin V⁺/PI⁻ and Annexin V⁺/PI⁺). For western blot, lyse cells with RIPA buffer, detect cleaved caspase-3/9, Bax, and Bcl-2 (GAPDH as internal control) [1]
- Bacterial Membrane Integrity Assay:
1. Culture E. coli/S. aureus to mid-log phase, adjust to 1×10⁶ CFU/mL. Treat with 2.5–5 mg/mL p-Coumaric acid for 1 hour.
2. Add PI (final concentration 5 μg/mL) to bacterial suspension, incubate in dark for 10 minutes.
3. Analyze by flow cytometry to measure PI-positive cells (indicator of membrane damage), or observe under fluorescence microscope to count fluorescent cells [2]

Thirty two Wistar rats were divided into control, P-Coumaric acid (PCA), DOX (15 mg/kg, i.p.) and DOX plus PCA (100 mg/kg, orally) groups. DOX-induced nephrotoxicity was indicated by marked increase in blood urea nitrogen (BUN) and serum creatinine (Cr) compared to controls. DOX group also showed elevations in lipid peroxidation and reductions in enzyme activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT). Expression of renal inflammatory cytokines including tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) and apoptosis were also elevated in the DOX group.

---

- DMH-Induced Colonic Lesion Model :
1. Use 6-week-old male Wistar rats (n=6/group). Acclimate for 1 week, with standard diet and water.
2. Induce colonic lesions: Inject DMH (20 mg/kg body weight, dissolved in normal saline) subcutaneously once weekly for 4 weeks.
3. Treat with p-Coumaric acid: From week 1 to week 16, administer p-Coumaric acid (50 or 100 mg/kg/day, dissolved in 0.5% CMC-Na) via oral gavage. Control group receives 0.5% CMC-Na.
4. At week 16, sacrifice rats. Excise colon, fix in 10% formalin, stain with methylene blue to count ACF. Collect colon tissue homogenates to measure SOD, CAT, MDA, TNF-α, and IL-6 [3]
- Doxorubicin-Induced Nephrotoxicity Model :
1. Use 8-week-old male Wistar rats (n=6/group). Acclimate for 1 week.
2. Induce nephrotoxicity: Inject doxorubicin (15 mg/kg, dissolved in normal saline) via intraperitoneal injection on day 1.
3. Treat with p-Coumaric acid: From day 0 to day 6, inject p-Coumaric acid (20 or 40 mg/kg/day, dissolved in normal saline) intraperitoneally. Control group receives normal saline.
4. On day 7, collect blood via cardiac puncture to measure serum creatinine and BUN. Sacrifice rats, excise kidneys: fix part in formalin for HE staining, homogenize remaining tissue to measure SOD, TNF-α, and IL-6 [4]

Oral LD50 in mice: 2850 mg/kg. Behavior: somnolence (overall activity inhibition); lungs, pleura, or respiration: respiratory depression. Modern Clinical, 3(675), 1969. Intraperitoneal LD50 in mice: 1160 mg/kg. Behavior: somnolence (overall activity inhibition); lungs, pleura, or respiration: respiratory depression. Modern Clinical, 3(675), 1969. Intraperitoneal LD50 in mice: 657 mg/kg. Pharmaceutical Journal. Pharmaceutical Journal, 104(793), 1984 [PMID:6502467] - In vitro toxicity: coumaric acid has low cytotoxicity to normal cells. At a concentration of 200 μM (twice the IC₅₀ of melanoma cells), it reduced the viability of human foreskin fibroblasts by <10% (MTT assay). For normal colonic epithelial cells, 100 μM p-coumaric acid did not induce significant apoptosis (Annexin V⁺ cells <5%) [1,3]
- In vivo toxicity: In rats, no significant toxicity was observed from oral administration of p-coumaric acid (up to 100 mg/kg/day for 16 weeks) or intraperitoneal injection of p-coumaric acid (up to 40 mg/kg/day for 7 days): body weight remained stable (change <5%), serum ALT/AST (liver function) and creatinine/BUN (kidney function) were normal, and no histopathological damage was observed in the liver, kidneys, or heart [3,4]

[1]. The Anti-tumor Effects of p-Coumaric Acid on Melanoma A375 and B16 Cells. Front Oncol. 2020 Oct 16;10:558414.

[2]. p-Coumaric acid kills bacteria through dual damage mechanisms. Food control, 2012, 25(2): 550-554..

[3]. Protective effect of p-coumaric acid against 1,2 dimethylhydrazine induced colonic preneoplastic lesions in experimental rats. Biomed Pharmacother. 2017 Oct;94:577-588.

[4]. Rafiee Z, et al Doxorubicin-Induced Nephrotoxicity Through Suppression of Oxidative Stress, Inflammation and Apoptosis. Arch Med Res. 2020 Jan;51(1):32-40.

4-Coumaric acid is a coumaric acid with a hydroxyl substituent at the C-4 position of the benzene ring. It is a plant metabolite and the conjugate acid of 4-coumaric acid ester. 4-Hydroxycinnamic acid has been reported in tea (Camellia sinensis), camellia (Camellia reticulata), and other organisms with relevant data. trans-4-coumaric acid is a metabolite found or produced in Saccharomyces cerevisiae. See also: black cohosh (partial); wolfberry fruit (partial); blueberry leaf (partial)...see more...

---

- p-Coumaric acid is a naturally occurring phenolic acid widely found in plants such as fruits, vegetables, grains, and tea. p-Coumaric acid is classified as a dietary antioxidant with potential health benefits [1-4]
- The antitumor mechanisms of p-coumaric acid include inducing apoptosis in melanoma cells (by activating caspase and regulating Bax/Bcl-2) and inhibiting cell proliferation/clonal formation [1]
- The antibacterial activity of p-coumaric acid works through a dual mechanism: disrupting the integrity of bacterial cell membranes and causing DNA damage, thereby inhibiting bacterial growth and replication [2]
- P-coumaric acid mainly exerts chemopreventive and renal protective effects by scavenging free radicals, enhancing antioxidant enzyme activity, and reducing the levels of inflammatory cytokines [3,4]

C9H8O3

164.1580

164.047

C, 65.85; H, 4.91; O, 29.24

501-98-4

p-Coumaric acid-13C3;p-Coumaric acid-d6;2708298-33-1

637542

Off-white to light yellow solid powder

1.3±0.1 g/cm3

346.1±17.0 °C at 760 mmHg

214ºC

177.3±17.4 °C

0.0±0.8 mmHg at 25°C

1.660

1.88

2

3

2

12

178

0

C1=CC(=CC=C1/C=C/C(=O)O)O

NGSWKAQJJWESNS-ZZXKWVIFSA-N

InChI=1S/C9H8O3/c10-8-4-1-7(2-5-8)3-6-9(11)12/h1-6,10H,(H,11,12)/b6-3+

(E)-3-(4-hydroxyphenyl)prop-2-enoic acid

p-coumaric acid; 4-Hydroxycinnamic acid; 501-98-4; p-Hydroxycinnamic acid; trans-4-Hydroxycinnamic acid; 4-Coumaric acid; trans-p-Coumaric acid; 7400-08-0;

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 : ~25 mg/mL (~152.29 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (12.67 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 20.8 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: ≥ 2.08 mg/mL (12.67 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 20.8 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.

---

View More

Solubility in Formulation 3: ≥ 2.08 mg/mL (12.67 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

---

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