Momordin Ic is identified as a novel SENP1 inhibitor [3].
SENP1 [3].

- In human hepatoblastoma HepG2 cells, Momordin Ic treatment (0-40 µM for 4-24 h) resulted in a dose- and time-dependent inhibition of cell proliferation. The IC50 values at 4h, 8h, 12h, and 24h were approximately 35 µM, 18 µM, 12 µM, and 7 µM, respectively [1].
Momordin Ic potently inhibited colony formation in HepG2 cells at concentrations of 5, 10, 15, and 20 µM [1].
Momordin Ic (15 µM, 4h) induced autophagy, as evidenced by an increase in MDC (monodansylcadaverine)-stained autophagic vacuoles and increased protein expression of Beclin 1 and LC3-II in a dose- and time-dependent manner in HepG2 cells [1].
Momordin Ic (15 µM, 4h) induced apoptosis, characterized by increased AO-EB staining, chromatin condensation, loss of mitochondrial membrane potential (MMP), increased Bax/Bcl-2 ratio, cytochrome c release, and activation of caspase-3 and caspase-8 in HepG2 cells [1].
The autophagy inhibitor 3-methyladenine (3-MA) inhibited apoptosis induced by Momordin Ic, while the autophagy activator rapamycin (RAP) enhanced apoptosis. Conversely, the apoptosis inhibitor ZVAD-fmk inhibited Momordin Ic-induced autophagy [1].
Momordin Ic (15 µM, 4h) increased reactive oxygen species (ROS) and H2O2 levels in HepG2 cells. Pretreatment with the antioxidant NAC inhibited both Momordin Ic-induced apoptosis and autophagy [1].
Momordin Ic (15 µM, 4h) suppressed the PI3K/Akt pathway. The PI3K/Akt inhibitor LY294002 increased both apoptosis and autophagy induced by Momordin Ic [1].
Momordin Ic (15 µM, 4h) modulated MAPK pathways; the ERK1/2 inhibitor U0126 inhibited autophagy, while the JNK inhibitor SP600125 and p38 inhibitor SB203580 inhibited both apoptosis and autophagy [1].
Momordin Ic (15 µM, 4h) inhibited NF-κB activation by suppressing IκB phosphorylation and NF-κB p65 nuclear translocation. The NF-κB inhibitor PDTC promoted apoptosis but inhibited autophagy [1].
[1]
- In human colon cancer cell lines HCT-8, HCT-116, SW480, and HT-29, Momordin Ic (2.5-20 µM for 24h) inhibited cell viability in a concentration-dependent manner. The IC50 values were approximately 13.03 µM, 10.67 µM, 17.5 µM, and 12.68 µM, respectively [3].
Momordin Ic (10 µM for 24h) increased the sub-G1 phase cell population and induced apoptosis in HCT-8 and HCT-116 cells [3].
Momordin Ic (10 µM for 24h) down-regulated c-Myc protein levels in a time-dependent manner without affecting c-Myc mRNA levels. It also suppressed the expression of c-Myc target genes (C23, GNL3, E2F2) and decreased protein levels of Bcl-2 while increasing Bax, cleaved Caspase-3, and cleaved Caspase-9 in HCT-116 cells [3].
Knockdown of c-Myc with siRNA abolished the anti-proliferative and pro-apoptotic effects of Momordin Ic in HCT-116 cells [3].
Momordin Ic (5-20 µM) increased the SUMOylation level of c-Myc (SUMO1 modification) in HCT-116 cells, an effect that was dependent on SENP1 [3].
Knockdown of SENP1 abolished the Momordin Ic-induced downregulation of c-Myc protein and the modulation of apoptosis-associated proteins (Bcl-2, Bax, Caspase-3, Caspase-9) in HCT-116 cells [3].
[3]
- In rats, pretreatment with Momordin Ic (30 mg/kg, p.o. for 14 days) significantly lowered serum AST, ALT, LDH, and γ-GT levels induced by carbon tetrachloride (CCl4). It also restored hepatic GSH levels and the activities of SOD, CAT, GSH-Px, GR, and GST, and reduced TBARS levels compared to CCl4-only treated rats [5].
Momordin Ic pretreatment (30 mg/kg, p.o. for 14 days) resulted in lower activities of the microsomal enzymes aminopyrine N-demethylase and aniline hydroxylase in CCl4-treated rats [5].
- In conscious mice, Momordin Ic (12.5, 25, and 50 mg/kg, p.o.) accelerated 30-minute gastrointestinal transit in a dose-related manner. The accelerations were 22.4%, 42.4%, and 51.9%, respectively, compared to control [4].
- In rats, oral administration of Momordin Ic (10 mg/kg) potentially inhibited ethanol-induced gastric mucosal lesions, reducing the lesion length by 87.3% and improving pathological changes [6].

- In a rat model of carbon tetrachloride (CCl4)-induced hepatotoxicity, Momordin Ic (30 mg/kg, p.o., once a day for 14 days) administered 30 minutes before CCl4 injection significantly reduced elevated serum levels of AST, ALT, LDH, and γ-GT. It also prevented the decrease in hepatic glutathione (GSH) levels and the reduction in activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR), and glutathione S-transferase (GST). Furthermore, it lowered hepatic malondialdehyde (MDA) levels and suppressed the increase in microsomal enzyme activities (aminopyrine N-demethylase and aniline hydroxylase) induced by CCl4 [5].
- In a rat model of ethanol-induced gastric mucosal lesions, Momordin Ic (10 mg/kg, p.o.) administered 60 minutes before 99.5% ethanol (1.5 mL/rat) showed a potent protective effect, reducing lesion length by 87.3% and improving pathological changes. This gastroprotective effect was attenuated by pretreatment with capsaicin (ablator of sensory nerves), L-NAME (NO synthase inhibitor), NEM (sulfhydryl blocker), or indomethacin (prostaglandin synthesis inhibitor) [6].
- In conscious fasted mice, Momordin Ic (12.5, 25, 50 mg/kg, p.o.) administered 60 minutes before a charcoal meal accelerated gastrointestinal transit in a dose-related manner. The effect (at 25 mg/kg) was attenuated by a single bolus of DL-p-chlorophenylalanine methyl ester (5-HT synthesis inhibitor), and by the 5-HT2 receptor antagonists cyproheptadine, ritanserin, and clozapine, but not by antagonists of 5-HT1, 5-HT3, or 5-HT4 receptors. Pretreatment with indomethacin (prostaglandin synthesis inhibitor) also attenuated the effect [4].

- Co-immunoprecipitation was used to assess the effect of Momordin Ic on c-Myc SUMOylation. HCT-116 cells were treated with Momordin Ic (5, 10, 20 µM) for 24 h. Cell lysates were immunoprecipitated with an anti-c-Myc antibody, and the immunoprecipitates were then analyzed by Western blot using antibodies against SUMO1, SUMO2, and SUMO3 to detect SUMOylated c-Myc [3].
[3]

- Cell viability was measured using the MTT assay. Cells were seeded in 96-well plates and treated with various concentrations of Momordin Ic for specified periods. Subsequently, MTT solution was added to each well and incubated for 4 h. The formazan crystals were then dissolved in DMSO, and the absorbance was measured at 490 nm using a microplate reader [1], [3].
- Cell colony formation was assessed by seeding cells in 6-well plates. After treatment with Momordin Ic for 2 weeks (with medium refreshed every 4 days), cells were fixed in methanol and stained with crystal violet. The colonies were then imaged and quantified [1], [3].
- Immunofluorescence staining was used to detect LC3 protein. HepG2 cells were fixed, permeabilized, blocked with goat serum, and then incubated with an LC-3 antibody overnight, followed by a secondary antibody. Cells were observed under a confocal fluorescent microscopy [1].
- Autophagic vacuoles were visualized using monodansylcadaverine (MDC) staining. After Momordin Ic treatment, cells were incubated with MDC solution. After washing, the cellular fluorescence was visualized using fluorescence microscopy [1].
- Apoptosis was assessed using AO-EB (acridine orange-ethidium bromide) and Hoechst 33258 staining. Treated cells were fixed and stained with AO-EB or Hoechst 33258, then observed under a fluorescence microscope for nuclear morphology changes [1].
- Mitochondrial membrane potential (MMP) was measured using the JC-1 probe. After treatment, cells were stained with JC-1 solution. The fluorescence intensity was measured using a microplate reader with settings for green (FL1) and red (FL2) fluorescence [1].
- Reactive oxygen species (ROS) levels were measured using the DCFH-DA assay. After treatment, cells were incubated with DCFH-DA. The DCF fluorescence intensity was measured by a fluorescence microplate reader at 485 nm excitation and 535 nm emission [1].
- Western blot analysis was performed to assess protein expression. Cells were lysed in RIPA buffer, and protein concentrations were determined. Protein samples were separated by SDS-PAGE, transferred to PVDF membranes, blocked, and incubated with primary antibodies. Membranes were then incubated with secondary antibodies, and proteins were visualized using an ECL kit [1], [3].
- Quantitative RT-PCR was performed to measure mRNA levels. Total RNA was isolated from cells using Trizol and reverse transcribed. Real-time PCR was carried out using SYBR Green master mix. The comparative CT (ΔΔCT) method was used to determine relative expression levels of genes like c-Myc, C23, GNL3, and E2F2 [3].
- Cell cycle progression was analyzed by flow cytometry. Cells were fixed in ice-cold ethanol, stained with propidium iodide (PI) and RNase A, and analyzed using a flow cytometer to determine the percentage of cells in G0/G1, S, and G2/M phases [3].
- Apoptosis rates were detected by flow cytometry using an Annexin V-FITC/PI apoptosis detection kit. Cells were stained with Annexin V-FITC and PI according to the kit's protocol and analyzed [3].
- Co-immunoprecipitation was performed to study protein-protein interactions. Cells were lysed, and lysates were precleared with protein A/G magnetic beads. The lysates were then incubated with an anti-c-Myc antibody or normal IgG, followed by incubation with protein A/G-agarose beads. The bead-bound proteins were eluted and analyzed by Western blot [3].
- For c-Myc protein stability assays, cells were treated with cycloheximide (CHX, a protein synthesis inhibitor) or MG132 (a proteasome inhibitor) in the presence or absence of Momordin Ic. c-Myc protein levels were then analyzed by Western blot at different time points [3].

- For hepatotoxicity study, male Sprague-Dawley rats (110-120 g) were orally administered Momordin Ic (30 mg/kg, dissolved in phosphate-buffered saline) once daily for 14 days. Thirty minutes after the final administration, a mixture of CCl4 in olive oil (1:1, v/v, 0.2 mL/100 g body weight) was injected to induce liver damage. The control group received saline. Animals were fasted for 12 hours before being sacrificed [5].
- For gastric lesion study, male Sprague-Dawley rats (about 250 g, fasted 24-26 h) were orally administered Momordin Ic (10 mg/kg, dissolved in phosphate-buffered saline) 60 minutes before oral administration of 99.5% ethanol (1.5 or 1.0 mL/rat) to induce gastric mucosal lesions. Control rats received the vehicle (PBS). One hour after ethanol administration, rats were sacrificed, and the stomachs were examined for lesions [6].
- For gastrointestinal transit study, male ddY mice (27-30 g, fasted 18-20 h) were orally administered Momordin Ic (12.5, 25, 50 mg/kg). Sixty minutes later, a charcoal meal (0.2 mL/mouse of a 1.5% CMC-Na and 5% charcoal solution) was given intragastrically. Thirty minutes after the charcoal meal, mice were sacrificed, and the distance traveled by the charcoal in the small intestine was measured [4].
- In the gastrointestinal transit study, various pretreatments were used. DL-p-chlorophenylalanine methyl ester (PCPA, 1000 mg/kg, p.o.) was given once at 1, 6, or 24 h before Momordin Ic; or twice (300 mg/kg x 2) at 72 and 48 h before Momordin Ic. Antagonists like methiothepin, cyproheptadine, ritanserin, clozapine, MDL 72222, metoclopramide, tropisetron, ketanserin, haloperidol, and indomethacin were injected subcutaneously 30 min before oral administration of Momordin Ic [4].

- Momordin Ic is rapidly eliminated in rats, characterized by a short elimination half-life (<2 h) [3].

[1]. Momordin Ic couples apoptosis with autophagy in human hepatoblastoma cancer cells by reactive oxygen species (ROS)-mediated PI3K/Akt and MAPK signaling pathways. Free Radic Biol Med. 2016 Jan;90:230-42.

[2]. Cytotoxic effects of recombinant proteins enhanced by momordin Ic are dependent on cholesterol and ganglioside GM1. Toxicon. 2023 Jun 15;229:107129.

[3]. Momordin Ic induces G0/1 phase arrest and apoptosis in colon cancer cells by suppressing SENP1/c-MYC signaling pathway. J Pharmacol Sci. 2021 Aug;146(4):249-258.

[4]. Acceleration of gastrointestinal transit by momordin Ic in mice: possible involvement of 5-hydroxytryptamine, 5-HT(2) receptors and prostaglandins. Eur J Pharmacol. 2000 Mar 24;392(1-2):71-7.

[5]. Roles of capsaicin-sensitive sensory nerves, endogenous nitric oxide, sulfhydryls, and prostaglandins in gastroprotection by momordin Ic, an oleanolic acid oligoglycoside, on ethanol-induced gastric mucosal lesions in rats. Life Sci. 1999;65(2):PL27-32.

[6]. Momordin Ic and oleanolic acid from Kochiae Fructus reduce carbon tetrachloride-induced hepatotoxicity in rats. J Med Food. 2005 Summer;8(2):177-83.

Momordicin Ic is a triterpenoid saponin. It has been reported that momordicin Ic exists in custard apple (Anredera baselloides) and heart-leaf custard apple (Anredera cordifolia), and relevant data are available for reference.

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- Momordin Ic simultaneously induces apoptosis and autophagy in HepG2 cells, and these two processes promote each other [1].
- The anticancer effects of Momordin Ic in HepG2 cells are regulated by ROS-mediated PI3K/Akt, MAPK (ERK, JNK, p38), and NF-κB signaling pathways [1].
- Momordin Ic induces cell cycle arrest and apoptosis in colon cancer cells by inhibiting SENP1-mediated c-Myc deSUMOylation, leading to reduced c-Myc protein stability [3].
- Momordin Ic accelerates gastrointestinal transit in mice, an effect that involves endogenous 5-HT, 5-HT2 receptors (possibly 5-HT2B and/or 5-HT2C subtypes), and prostaglandins [4].
- The gastroprotective effect of Momordin Ic against ethanol-induced gastric lesions in rats involves capsaicin-sensitive sensory nerves, and endogenous nitric oxide, sulfhydryls, and prostaglandins [6].
- Momordin Ic and oleanolic acid from Kochiae Fructus reduce carbon tetrachloride-induced hepatotoxicity in rats by enhancing the hepatic antioxidant defense system [5].

C41H64O13

764.9391

764.434

96990-18-0

176596

White to off-white solid

1.4±0.1 g/cm3

886.2±65.0 °C at 760 mmHg

263.1±27.8 °C

0.0±0.6 mmHg at 25°C

1.606

9.87

7

13

6

54

1510

17

C[C@@]12CC[C@H]3C([C@H](CC[C@]3(C)[C@H]1CC=C1[C@@H]3CC(C)(C)CC[C@@]3(CC[C@@]21C)C(=O)O)O[C@@H]1O[C@H](C(=O)O)[C@@H](O)[C@H](O[C@@H]2OC[C@@H](O)[C@H](O)[C@H]2O)[C@H]1O)(C)C

HWYBGIDROCYPOE-WEAQAMGWSA-N

InChI=1S/C41H64O13/c1-36(2)14-16-41(35(49)50)17-15-39(6)20(21(41)18-36)8-9-24-38(5)12-11-25(37(3,4)23(38)10-13-40(24,39)7)52-34-29(46)30(28(45)31(54-34)32(47)48)53-33-27(44)26(43)22(42)19-51-33/h8,21-31,33-34,42-46H,9-19H2,1-7H3,(H,47,48)(H,49,50)/t21-,22+,23-,24+,25-,26-,27+,28-,29+,30-,31-,33-,34+,38-,39+,40+,41-/m0/s1

(2S,3S,4S,5R,6R)-6-[[(3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-carboxy-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-yl]oxy]-3,5-dihydroxy-4-[(2S,3R,4S,5R)-3,4,5-trihydroxyoxan-2-yl]oxyoxane-2-carboxylic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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

DMSO : ~100 mg/mL (~130.73 mM)

Solubility in Formulation 1: ≥ 6.25 mg/mL (8.17 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 62.5 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: ≥ 2.5 mg/mL (3.27 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (3.27 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 25.0 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.)