Table 3.
Metabolism Studies (Metabolite Profiling) of Herb-Derived Compounds with Anticancer Activities
| Anticancer Drug | Type of Study | Biochemical Tool/Animal/Human (Route: Dose) | Type of Sample | Analytical Technique | Outcome | Ref. | ||
|---|---|---|---|---|---|---|---|---|
| Metabolite | Metabolic Pathway | Efficacy/Toxicity of Metabolite | ||||||
| Alantolactone (Inula helenium L.) | In vivo | Male Sprague–Dawley rats (PO: 100 mg/kg) | Urine, feces, bile | UPLC-TOF-MS | 11 Metabolites for urine (M1-M5, M10-M11, M14-M16, M18); 10 Metabolites for bile (M1-M4, M8, M12, M25, M33, M43-M44); 38 Metabolites for feces (M1-M13, M17-M25, M27-M31, M33, M35-M44) |
Oxidation (M1-M9); Di-oxidation (M10-M11); Demethylation (M12-M13); Demethylation to carboxylic acid (M14-M15); Hydration (M16); Hydrogenation (M17-M18); Addition of H2S (M19-M20); Addition of H2S + oxidation (M21-M23); AL2-S (M24-M25: one sulfur-containing dimer metabolites of alantolactone); AL2-S + oxidation (M27-M31); AL2-SS (M33: two sulfur-containing dimer metabolites of alantolactone); AL2-SS + oxidation (M35-M39); AL2-SSS (M40: three sulfur-containing dimer metabolites of alantolactone); AL2-SSSS (M41: four sulfur-containing dimer metabolites of alantolactone); AL2-SSSSS (M42: five sulfur-containing dimer metabolites of alantolactone); Cysteine conjugation (M43); N-acetylcysteine conjugation (M44) |
Not evaluated | [9] |
| Astragali Radix water extract (containing calycosin-7-β-glucoside, formononetin, calycosin, ononin, astragaloside IV) | In vivo | Male Sprague–Dawley rats (PO: 4 g/kg and 16 g/kg Astragali Radix water extract) | Plasma | UHPLC-MS/MS | 4 Metabolites (calycosin-7-β-glucoside-3ʹ-glucuronide for calycosin-7-β-glucoside, formononetin-3ʹ-glucuronide for formononetin; calycosin-3ʹ-glucuronide for calycosin, daidzein-3ʹ-glucuronide); No metabolites for ononin and astragaloside |
Glucuronidation | Not evaluated | [71] |
| Calphostin C (Cladosporium cladosporoides) | In vitro | CD-1 MLM | – | LC-MS | 1 Metabolite (after incubating liver microsomes with porcine esterase) | Breaking the ester bond | Not evaluated | [72] |
| In vivo | Female CD-1 mice (IP: 40 mg/kg) | Plasma | 1 Metabolite | |||||
| CAT (3,6,7-trimethoxyphenan-throindolizidine (isolated from Tylophora atrofolliculata)) | In vivo | Male Wistar rats (PO: 6 mg/kg) | Urine | RRLC-ESI-QTOF-MS | 21 metabolites (M1-M21) | Di-demethylation + di-glucuronidation (M1-M3); Di-demethylation + glucuronidation (M4-M9); Demethylation + glucuronidation (M10-M12); Di-demethylation (M13-M15); Demethylation (M16-M18); Oxidation (M19); Hydroxylation (M20-M21) |
Not evaluated | [73] |
| Dimethoxycurcumin (Curcumin analog) | In vitro | MLM (male CD-1), HLM (CYPream) | – | HPLC-QTRAP, HPLC-QqQ-LIT-MS | 8 Metabolites for MLM (369, 371, 383, 385, 399, 401, 559, 561); 7 Metabolites for HLM (369, 383, 385, 399, 401, 559, 561) |
Di-O-demethylation (369); Di-O-demethylation + hydrogenation (371); O-demethylation (383); O-demethylation + hydrogenation (385); Hydrogenation (399); Di-hydrogenation (401); O-demethylation + glucuronidation (559); O-demethylation + hydrogenation + glucuronidation (561) |
Not evaluated | [74] |
| Fisetin (flavonoid compound) | In vivo | Female C57BL/6J mice (IP: 223 mg/kg) | Plasma | HPLC-MS/MS | 3 Metabolites (M1, M2, M3: geraldol) | Glucuronidation (M1, M2); Methoxylation (M3) | M3: 2.5 and 1.1-fold higher cytotoxic effect against LLC cell line (Lewis carcinoma) and EAhy 926 cell line (endothelial cell) compared to fisetin, respectively; M3 exhibited 1.5-fold fold higher cytotoxic effect against NIH 3T3 cell line (normal cell) |
[75] |
| Flavone-8-acetic acid (FAA) | In vitro | HLM, MLM expressing CYP enzyme induced by Aroclor 1245 | – | HPLC-UV, HPLC-UV-MS | 6 Metabolites for MLM (M1: 3ʹ,4ʹ-dihydrodiol-FAA, M2: 5.6-epoxy-FAA, M3a: 4ʹ-OH-FAA, M3b: 3ʹ-OH-FAA, M3c: 3ʹ,4ʹ-epoxy-FAA, M4: 6-OH-FAA); One human liver microsome (sample 14) produced M3a and another human liver microsome (sample 15) produced 3 metabolites of M1, M3a, M3c: 3ʹ,4ʹ-epoxy-FAA |
Epoxide hydrolase reaction (M1); Epoxidation (M2, M3c); Hydroxylation (M3a, M3b, M4) |
Interspecies difference metabolism could involve with difference anticancer activity | [76] |
| Furanodiene (Rhizoma Curcumae) | In vitro | Rat liver S9, RLM | – | HPLC-ESI-MS, HR-ESI-MS, and 1H NMR, 13C NMR, 2D NMR | 6 Metabolites for rat liver S9 (M1: 1β,10α,4α,5β-diepoxy-8α-hydroxy-glechoman-8α,12-olide, M2: 2β-hydroxyl-aeruginolactone, M3: 14-hydroxyl-aeruginolactone, M4: 1β,8β-dihydroxyeudesm-4(14),7(11)-dien-8α,12-olide or 1β,8β-dihydroxyeudesm-4,7(11)-dien-8α,12-olide, M5: 1β,8β-dihydroxyeudesm-3,7(11)-dien-8α,12-olide M6: aeruginolactone); 1 Metabolite for RLM (M6) |
Oxidation (M6); Oxidation + Hydroxylation (M2, M3); Epoxidation (M1); Epoxidation + new bond formation (M4, M5) |
Not evaluated | [77] |
| In vivo | Male Sprague-Dawley Rat (PO: 100 mg/kg) | Bile, urine, feces | 6 Metabolites for bile and urine (M1-M6); 1 Metabolite for feces (M6) |
|||||
| (-)-grandisin (extracted from Piper solmsianum) | In vitro | HLM | – | GC-MS and LC-MS | 4 Metabolites (M1: 4-O-demethylgrandisin, M2: 4,4′-di-O-demethylgrandisin, M3: 3,4-di-O-demethylgrandisin or 3,5-di-O-demethylgrandisin, M4: 3-O-demethylgrandisin) | Demethylation | Not evaluated | [2] |
| Irisflorentin (Belamcanda chinensis) | In vitro | RLM | – | HPLC-UV | 7 Metabolites (M1: 6,7-Dihydroxy-5,3′,4′,5′-tetramethoxy isoflavone, M2: Irigenin, M3: 5,7,4′-Trihydroxy-6,3′,5′ trimethoxy isoflavone, M4: 6,7,4′-Trihydroxy-5,3′,5′-trimethoxy isoflavone, M5: 6,7,5′-Trihydroxy-5,3′,4′-trimethoxy isoflavone; M6 and M7: unidentified |
Cleavage of methylene acetyl group (M1); Adding of hydroxyl and methoxyl groups (M2-M5) |
M1 & M2: 4.6 and 8.4-fold cytotoxic activities against DU145 and 4- and 8.2-fold against MCF-7 cell lines compared with parent compound. | [78] |
| Leelamine (bark of pine tree) | In vitro | HLM | – | LC-MS/MS | 1 Metabolite in the presence of NADPH generation system | Hydroxylation | Not evaluated | [7] |
| In vivo | Male ICR mice (IP: 10 mg/kg) | Urine, feces | 1 Metabolite for urine not in feces | |||||
| MPD (Methyl protodioscin: isolated from Dioscorea collettii var. hypoglauca)) | In vivo | Male Sprague-Dawley rats (PO: 80 mg/kg) | Urine | 1H NMR, 13C NMR, HRSI-MS | 10 Metabolites (M1: Dioscin, M2: pregna-5,16-dien-3β-ol-20-one-O-α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-β-D- glucopyranoside, M3: diosgenin, M4: protobioside, M5: methyl protobioside, M6: 26-O-β-D-glucopyrannosyl (25R)-furan-5-ene-3β,22α, 26-trihydroxy-3-O-α-L-rhamnopy-ranosyl-(1→ 4)-β-D-glucopyranoside, M7: 26-O-β-D-glucopyranosyl (25R)-furan-5-ene-3β,26-dihydroxy-22-methoxy-3-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranoside, M8: prosapogenin A of dioscin, M9: prosapogenin B of dioscin, M10: diosgenin-3-O-β-D-glucopyranoside) |
Dealkylation; Dehydration; Oxidation |
All metabolites: lower cytotoxic activities against human HepG2, NCI-H460, MCF-7 and HeLa cell lines than parent drug & MPD. However, M1 and M4 exhibited strong anti-cancer activities against HepG2 (M1), NCI-H460 (M1, M4), and HeLa cell lines (M4). | [79] |
| Oridonin (ORI) (Diterpinoid of Isodon rubescens) | In vivo | Male Spraque-Dawley rats (PO: 10 mg/kg) | Bile, urine | UPLC-QqQ TOF-MS | 17 Metabolites for bile (M1-M4, M6-M18); 10 Metabolites for urine (M1-M6, M12, M14-M16) |
Hydroxylation (M1-M3); Ketone Formation (M4); Hydroxylation + hydration (M5); Dehydroxylation (M6-M7); Di-dehydroxylation (M8-M11); Dehydration (M12); Dehydroxylation + dehydration (M13); Didehydroxylation + oxidized to carboxylic acid (M14); Desaturation + oxidized to carboxylic acid (M15); Hydration (M16); Dehydroxylation + glucuronidation (M17, M18) |
Not evaluated | [80] |
| Quercetin (flavonoid) | In vitro | Human hepatocellular carcinoma cell lines (HepG2) | – | HPLC-RD | 1 Metabolite (4) | O-methylation | Not evaluated | [81] |
| Trabectedin (ET-743: isolated compound from Ecteinascidia turbinata) | Clinical trial | Cancer patients (IV: 1 mg of trabectedin (2.5 MBq [14C] trabectedin (70 µCi))) | Urine and feces | HPLC-QqQ-MS and LC-LSC | 8 Fractions for urine (U1-U8); 10 Fractions for feces (F1-F10) (U4, F3, and F5: ETM-204; U5 and F6: ETM-217; U6 and F7: ET-759A and ET-731; U7 and F9: ET-745, ETM-259; U8 and F10: ET759B) |
Dehydroxylation (ET-745); Dehydroxylation +demethylation (ET-731); Oxidation (ET-759A); Breaking up of the molecule to the individual tetrahydroisoquinoline subunits (ETM259 and ETM204); ETM-259 + acetate ester hydrolysis (ETM217) |
Not evaluated | [82] |
| Yuanhuapine (Isolated from Daphne genkwa) | In vivo | Male Sprague-Dawley rats (Oral: 5 mg/kg) | Urine | UPLC-QTOF-MS | 12 Metabolites (M1-M12) | Hydroxylation (M1-M4); Di-hydroxylation (M5, M6); Tri-hydroxylation (M7); Hydroxylation + glucuronidation (M8); Methylation + di-hydroxylation (M9); (Hydroxylation + methylation) + (hydroxylation + glucuronidation) (M10); Reduction + 2(Cysteine conjugation) (M11); Hydration + (S-Cysteine conjugation) (M12) |
Not evaluated | [3] |
| Ziyuglycoside II (Sanguisorba officinalis L.) | In vitro | Rat liver homogenate | – | LC-QTOF-MS | 16 metabolites (H-M1-H-M16) | Glucuronidation (H-M1); Glucose conjugation (H-M2-H-M4); Dehydration + glucuronidation (H-M5); Formolation of ziyuglycoside II (adding CO: H-M6); Oxidation + reduction (H-M7); Dehydrogenation (H-M8); Demethylation (H-M9); Dehydration (H-M10, H-M11); Dearabinosylation + oxidation (H-M12, H-M13); Dearabinosylation (H-M14); Dearabinosylation + reduction (H-M15, H-M16) |
Not evaluated | [83] |
| In vivo | Male Sprague-Dawley rats (Oral: 50 mg/kg) | Urine, feces | UFLC-QTOF-MS | 10 Metabolites for urine (U-M1-U-M10); 10 Metabolites for feces (F-M1 and F-M10) |
Glucuronidation + glycosylation (U-M1); Glucuronidation + deglycosylation + decarboxylation + dihydroxylation (U-M2); U-M2 + deoxidation + methylation (U-M3); Glucuronidation + loss of C2H2O (U-M4); U-M4 + deoxidation + methylation (U-M5); Decarboxylation (U-M7); Glucosylation (U-M6); Dearabinosylation (U-M9 and F-M9); Dearabinosylation + oxidization (U-M8, F-M5, F-M6); Dearabinosylation +dehydrogenation (U-M10); Oxidation (F-M1); F-M1 + dehydrogenation (F-M2); Dehydration (F-M3and F-M4); Dearabinosylation + loss of CO (F-M7); Dearabinosylation + demethylation (F-M8); F-M9 + dehydrogenation (F-M10) |
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Abbreviations: ESI, Electrospray Ionization; GC, Gas Chromatography; HLM, Human Liver Microsome; HPLC, High Performance Liquid Chromatography; HR, High Resonance; HRSI, High Resolution Single Ion; IP, Intraperitoneal; IV, Intravenous; LC, Liquid Chromatography; MS, Mass Spectrometry; MLM, Monkey Liver Microsome; NMR, Nuclear Magnetic Resonance spectroscopy; PO, Per oral; QqQ, Triple quadrupole; LIT, Linear Ion Trap; LSC, Liquid Scintillation Counting; QTOF, Quadrupole Time-of-flight; QTRAP, Linear ion trap Triple Quadrupole; RD, Radio Detection; RLM, Rat Liver Microsome; RRLC, Rapid Resonance Liquid Chromatography; TIS, Turbo Ion Spray; TOF, Time-of-Flight; UHPLC, Ultra-high performance liquid chromatography; UPLC, Ultra Performance Liquid Chromatography; UV, Ultraviolet detection.