Evodia rutaecarpa

Evodia Rutaecara, the dried unripe fruit of which is also known as Wu zhu yu (Wu Zhu Yu, interchangeably) or Evodia Fruit, is a herb used in Traditional Chinese Medicine at doses of 3-9 grams (of the berries) thrice a day for the purposes of warmth, intestinal comfort (specifically; to alleviate abdominal pain, acid regurgitation, nausea and diarrhea), dysmenorrheal, and fighting inflammation and infections.^[1] It is frequently used in a combination supplement called Wu Zhu Yu Tang, which consists of Evodia fruit with Jujube Fruit and Panax Ginseng (1:1:1 ratio) and Ginger root (twice the amount of any one other ingredient) which is a decoction used for hypertension or as Zuo jin wan which is used for gastrointestinal distress alongside Rhizoma Coptidis at a 1:6 ratio (Evodia:Coptidis). Another decoction exists called Fan zuo jin wan, which is Rhizome Coptidis and Evodia in the inverse ratio.

When consuming Wu Zhu Yu Tang in a traditional manner, the estimated intake of Rutaecarpine is 16mg daily (extrapolated from the average intake of 9g berry extract taken thrice a day) and slightly higher levels of evodiamine.^[2][3]

Traditionally eaten when you are cold; apparently warms you up real nicely while making your stomach and intestines happy. Also supposedly fights cancer

• Evodiamine^[4] and dehydroevodiamine, two forms of one of the main compounds belong to the 'quinazolinocarboline alkaloid' class ^[5] Evodiamine is found in the range of 0.2-1.6%, with the lower range of the samples tending to have more dehydroevodiamine (0.12-0.86%), suggesting interconversion^[6]
• Rutaecarpine and its metabolite 10-hydroxyrutaecarpine (as well as the former's glycoside, rutaecarpine-10-O-rutinoside^[7]) the forms of the other main quinazolinocarboline alkaloid.^[5] Rutaecarpine is at around 0.15-0.55% in evodia fruit.^[6]
• Other quinazolinocarboline alkaloids such as wuchuyine and rhetsinine^[8][9] and Evodiamide, which is at around 0.005-0.08% usually, although some plants have been noted as high as 0.4-0.6%^[6]
• The quinoline alkaloids Evocarpine, Dihydroevocarpine, 1-methyl-2-n-nonyl-4(1H)quinolone^[5] as well as a large amount (17+) of others^[10]
• Wuzhuyuamide-I^[7] at low levels (0.00003%)^[11]
• Flavonoids and flavonoid glycosides such as isorhamnetin-7-O-rutinoside and diosmetin-7-O-β-d-glucopyranoside^[5]
• Acylgluconic acids such as trans-feruloylgluconic acid and trans-caffeoylgluconic acid^[11] at 0.00003% and 0.0006% whole fruit, respectively (although can be isolated in a methane-ethanol extraction)^[11]
• Limonoid compounds such as Limonin^[12] (not to be confused with Limonene) as well as evodirutaenin, evodol, and shihulimonin A^[13]
• Myo-inositol^[11] at 8g per 299.5g sample (2%)^[11]
• Pthalic acid dibutyl ester at 8mg per 299.5g (miniscule)^[11]
• Essential oils of β-pinene (72.82%), 1R-α-pinene (8.90%), and β-myrcene (1.99%)^[14]

Main collection of compounds are the quinazolinocarboline alkaloids, evodiamine and rutaecarpine (named after the two parts of the herbs name respectively, Evodia Rutaecarpa). The flavonoids, limonoid, and other alkaloid compounds are most likely also bioactive, and the Inositol content is quite respectable and may be bioactive. The content of the main bioactives is relatively low, however

Pictured below are the two main bioactives typically attributed to Evodia Rutaecarpa.^[15]

Despite the similarities in action to capsaicin, which is hot pepper extract; isolated evodiamine possesses no significant taste nor spiciness. A large amount of crystal data for evodiamine can be found here.^[4]

After oral administration of a Wu zhu yu decoction (Evodia, Panax Ginseng, and Jujubae Fructus at a 1:1:1 ratio, and Ginger at twice the amount of any one ingredient; ethanolic extract) given at 12g/kg orally (2.4g/kg Evodia ethanolic extract) the following parameters were found for various compounds in Evodia Rutaecarpa.^[5]

• Evodiamine had a half-life of 0.93 ± 0.45 hours, a T_max of 1.49 ± 0.22 hours, a C_max of 19.52 ± 8.17ug/mL, and an AUC of 71.27 ± 15.52ug/h/mL.^[5] Another study finding a C_max of 49 ± 19ng/mL after 500mg/kg ingestion evodiamine in isolation,^[16] along with the former pinpoint a bioavailability of pure evodiamine at around 0.1% in isolation and slightly higher as Evodia Rutaecarpa. Another study using basic Evodia ethanolic extract found that 40mg/kg of an extract (35% evodiamine by weight, so 14mg/kg) conferred a C_max of 164.8 ± 65.1ug/mL;^[17] relative to the previous study, this suggests that a dose of 35.7-fold lower, if taken in an ethanolic berry extract, has an average peak blood concentration 3.36-fold higher. The T_max in all studies seems to hover between 30-60 minutes, however.^[16][17][5]

• Rutaecarpine had a half-life of 0.98 ± 0.39 hours, a T_max of 1.26 ± 0.23 hours, a C_max of 13.46 ± 7.03ug/mL, and an AUC of 62.44 ± 18.85ug/h/mL.^[5] Another study found low bioavailability of Rutaecarpine in isolation (Concentration of 2.4 ± 3.0ng/ml 30 minutes after 40mg/kg crude drug^[18]) but administering Rutaecarpine as a solid dispersion (mechanical technique) increased the concentration in the blood 7.5-fold (to 18.1 ± 1.8ng/mL^[18]) Nanoemulsions have also shown benefit in increasing the absorption rates.^[19] A basic concentration ethanolic extract of Wu Zhu Yu (pre-ripe) berries (40% of a 40mg/kg oral ingestion, so 16mg/kg), reaches blood levels of 215.3 ± 80.4ug/mL in rats.^[17] The improvement in bioavailability is similar to that seem with evodiamine, where an oral dose 2.5-fold lower can have a blood concentration 89.7-fold higher if consumed in the form of a concentrated berry ethanolic extract. Similar to evodiamine, T_max of all studies hovered around 30-60 minutes regardless of the C_max.^[5][18][17]

• Dehydroevodiamine had a half life of 0.79 ± 0.21 hours, a T_max of 1.07 ± 0.15 hours, a C_max of 7.64 ± 0.63ug/mL, and an AUC of 12.39 ± 2.71ug/h/mL.^[5] In isolation, dehydroevodiamine appears to have better bioavailability than the previous two molecules (averaged at 15.35% in rats^[9]) yet at least one study has investigated Dehydroevodiamine and its circulating levels in conjunction with other plants, and consumption of Evodia alongside Rhizoma Coptidis (1:6 ratio, combination known as Zuojinwan) appears to preserve the T_max (1.6 hours without, 1.8 hours as Zuojinwan) yet elevate the C_max 2.66-fold (15,383 ± 7166 to 40,992 ± 21,052) while reducing the T_max (3.5 ± 3.0 hours to 1.5 ± 1.1) and causing a large increase in the AUC to infinity by 174% (68,134 ± 19,162 to 186,715 ± 39,211).^[20]

The three main components of Evodia Rutaecarpa all appear to be highly subject to herb-herb interactions, where their absorption and circulating levels are much higher when in the form of the whole plant rather than isolated chemical and even higher when paired in Traditional Chinese Medicine decoctions. It may be worthless to supplement evodiamine or rutacarpine in isolation if your goal is to get it to your blood (fine if you want it in the colon) due to poor bioavailability

• 10-hydroxyrutaecarpine had a C_max of 0.76 ± 0.16ug/mL and a T_max of 0.50 ± 0.25 with an AUC of 9.32 ± 2.93ug/h/mL, but an undetectable half-life.^[5]
• 1-methyl-2-n-nonyl-4(1H)quinolone had a C_max of 3.16 ± 1.28ug/mL at a T_max of 0.77 ± 0.15 hours, its AUC was 9.83 ± 1.51ug/h/mL and half-life was 2.18 ± 0.47 hours.^[5]
• Evocarpine had a half-life of 0.53 ± 0.18 hours and a T_max of 0.88 ± 0.17 hours, with a C_max of 11.53 ± 6.97ug/mL and an AUC of 33.66 ± 10.52ug/h/mL.^[5]
• Dihydroevocarpine had a half-life of 0.49 ± 0.21 hours and a T_max of 0.79 ± 0.15 hours, a C_max of 6.05 ± 2.87ug/mL and an AUC of 16.53 ± 5.79ug/h/mL.^[5]
• Isorhamnetin-7-O-rutinoside had a half-life of 0.67 ± 0.30 hours, a T_max of 0.95 ± 0.25 hours, a C_max of 2.21 ± 0.32ug/mL, and an AUC of 7.54 ± 1.03ug/h/mL.^[5]
• Diosmetin-7-O-β-d-glucopyranoside had a half-life of 0.95 ± 0.51 hours, a T_max of 1.53 ± 0.17 hours, a C_max of 1.73 ± 0.51ug/mL, and an AUC of 9.41 ± 3.57ug/h/mL.^[5]

In general, all compounds tend to have low to moderate bioavailability and relatively rapid pharmacokinetic profiles; hitting their peak concentrations in the blood before or at an hour after administration. Evodiamine and Rutaecarpine seem to really benefit from being ingested as a whole berry rather than isolated compounds

Evodiamine appears to be carried in the blood and distributed into organs, and one study in rats found the volume of distribution to be 560ml/kg^[21] and found evodiamine (and metabolites) to be deposited in the liver, kidneys, heart, lung, and adipose tissue at a concentration higher than plasma, and other tissues it diffused in were at lower levels relative to plasma.^[21]

When investigating dehydroevodiamine, it has been demonstrated that this molecule can cross the blood brain barrier of the rat and enter the brain via linear kinetics.^[22] When a plasma level of 4.82 ± 1.55 μg/mL was measured in rats, the concentrations in the brain were 1.11 ± 0.4 (cortex), 0.93 ± 0.24 (hippocampus), 0.64 ± 0.28 (striatum), 1.13 ± 0.33 (cerebellum), 1.04 ± 0.3 (brain stem), and 1.18 ± 0.18 μg/g (everything else).^[9] On average, concentrations of dehydroevodiamine that circulate in the brain are 3-4x lower than plasma levels.^[9]

Dehydroevodiamine appears to be subject to P450, and its circulating metabolites consist of five glucuronides (two identified ones at the 10 and 11 carbon) and one sulfate (Dehydroevodiamine-12-sulfate) in freely moving rats.^[9]

Rutaecarpine is metabolites mostly by cytochrome P450 1A2 (aromatase) at the 10, 11, and 12 carbon positions, although 3 is not unheard of^[23][24] and into the four metabolites of 3, 10, 11, or 12-hydroxyrutaecarpine.^[25] Metabolism into 3- and 10- hydroxyrutaecarpine is strongly inhibited by ketoconazole, suggesting it is metabolized by CYP3A4; the other two metabolites (11- and 12-hydroxyrutaecarpine) are metabolized by CYP1A but can also be metabolized by CYP3A4 and CYP2D6.^[3]

Interestingly, Rutaecarpine's metabolite 10-hydroxyrutaecarpine, which is produced by CYP3A4, can act as an aromatase inhibitor, inhibiting CYP1A1 and CYP1A2 (two aromatase isomers) with IC₅₀ values of 2.56 ± 0.04uM and 2.57 ± 0.11uM, respectively.^[3] Rutaecarpine also shares aromatase inhibitory potential with preference for CYP1A2,^[26] but prolonged (3 days) ingestion of rutaecarpine,^[27] Evodia,^[2] or a Wu Zhu Yu Tang mixture of which effects are attributable to Evodia^[2] may cause CYP1A induction and thus the opposite effects of inhibition.

Rutaecarpine technically is an aromatase inhibitor, but causes increased aromatase activity relatively quickly

CYP1B1 is also inhibited by 10-HRT at 0.09 ± 0.01uM.^[3] CYP3A4 seems to be unaffected by rutaecarpine.^[27]

24 hours after administration of evodiamine, 82% of evodiamine and its metabolites are excreted with 23% of that excretion occuring via the urine and the rest via feces.^[21]

After an oral bolus of 500mg/kg bodyweight dehydroevodiamine, most of it is conjugated by P450; the amount of unchanged metabolite in the urine and feces are 0.5% and 6% of the initial oral dose, respectively.^[9]

Rutaecarpine has a large amount of fecal excretion (~42%) after oral ingestion, and its main urinary metabolite is 10-hydroxyrutaecarpine, which is the result after rutaecarpine interacts with the CYP1A2 enzyme.^[25]

Evodiamine is an agonist of the vanilloid receptor,^[28] a property also seen with the red pepper extract capsaicin. It appears to be slightly less potent than capsaicin, requiring thrice the concentration in vitro to maximally stimulate the receptors.^[28][29]

Through interactions with vanilloid receptors, Evodia Rutacarpa possesses antinocioreceptive (pain relieving) effects.^[30]

Evodia Rutacarpa has been investigated for its effects on body fat due to it being used traditionally as a warming agent and as a 'hot herb'; references to thermogenesis in the Chinese literature.

1-3mg/kg bodyweight evodiamine administered subcutaneously was able to drop core body temperature by 1C in fasted mice, while fed mice required 10mg/kg to achieve the same effect.^[31] These drops in internal temperature were matched with extra heat dissipation from the rat tail, indicative of thermogensis, almost immediately.^[31] The ability of evodiamine's mechanisms of action, TRPV1 activation, to make areas tolerant to the cold is also a possibility; cold hyposensitivty (responding less to the cold) indirectly increases perceptions of heat.^[32]

This heat production, however, is not the sole reason for evodiamine's fat burning effects.^[33]

Evodiamine has been traditionally used to increase warmth, and it appears it might increase both heat production and reduce the perception of cold. However, the studies from which this data is drawn are not suited to human oral consumption of Evodia

When incubated in preadipocytes, evodiamine is able to activate the MAPK cascade, which reduces insulin-induced phosphorylation of Akt and also PPARγ activity, which thus decreases preadipocyte differentiation.^[33] The classical mechanisms of evodiamine, as agonist of TRPV1 receptors,^[34] can also work to reduce preadipocyte differentitation;^[35] thus there may be two similar mechanisms of action occurring.

Inhibition of preadipocyte differentiation has been noted elsewhere relatively dose-dependently, but most significantly at 4uM concentration or above^[36] and in vivo following injections of evodiamine.^[37] The authors mentioned this was 'negative crosstalk' with insulin signalling, which interferes with its effects.^[33]

Evodiamine appears to exert anti-obesity effects via inhibiting preadipocyte differentiation

One study on mice fed 0.03% evodiamine and rats fed 1.35% evodia extract (standardized to 0.02% evodiamine) in their obesity causing diets over 21 days showed no difference in food intake compared to control (important to note, as the adverse taste of capsaicin screws with food intake in animal studies) but a small reduction in fat mass on mice (28% reduction of perirenal fat, 11% less epididymal fat) and decreases in weight (-10.3% relative to control) and increased thermogenesis; suggesting evodiamine may exert an anti-obesity effect.^[31] Another study investigating mechanisms fed 0.03% evodiamine for 6 months and noted significant anti-obesity effects by reducing the rate of weight gain, and this persisted in UCP1 knockout mice who were unable to produce heat from evodiamine.^[33]

Evodiamine has been shown to prevent fat gain to a degree in mice fed a diet that induces fat gain, but has not yet been demonstrated to induce fat loss. The heat producing effects of evodiamine may not be related to attenuation of fat gain

When evodiamine is incubated with endothelial cells, it is able to inhibit IL-1α (at 10uM) and Thromboxane B2 (TXB2) secretion (at 10uM) in response to inflammatory signals (LPS) and was able to decrease the levels of E-selectin on endothelial cells; although no response was dose-dependent.^[38]

Both evodiamine and rutaecarpine have been shown to inhibit PGE(2) production in macrophages that are stimulated by LPS (a pro-inflammatory signal)^[39] and in macrophages undergoing hypoxia.^[40] Evodiamine was further able to prevent upregulation of COX-2, a pro-inflammatory enzyme, while rutaecarpine was ineffective at doing so.^[39] Dehydroevodiamine can also reduce COX-2 activity and mRNA translation^[41] while rutaecarpine seems to be more of a direct inhibitor rather than manipulating protein content of COX enzymes, able to inhibit both COX-1 and COX-2 with IC₅₀ values of 8.7uM and 0.28uM, respectively.^[42]

Another compound, goshuyuamide II, was demonstrated to be able to inhibit 5-LOX and reduce synthesis of leukotrienes.^[39]

The combination decoction of Evodia Fructae and Coptidis Chinensis 1:6 (Zuo Jin Wan) has been shown in vitro to inhibit both NF-kB translocation and AP-1 signalling in HepG2 (liver) cells with IC₅₀ values of >200 and 22.9μg/ml for AP-1 and NF-kB, respectively for the combination.^[14] Berberine was able to inhibit AP-1 and NF-kB with IC₅₀ values of 9.5 and 50μM, respectively, while Evodiamine was only inhibitory on AP-1, and to lesser potency than Berberine.^[14]

Evodiamine has been shown to inhibit NF-kB activation in macrophages at 1-10uM concentration in response to pro-inflammatory signals.^[39]

Topoisomerases (I and II) are enzymes that regulate the separating of DNA strands so DNA can be replicated. DNA gets unwound when a phenolic nucleophile attacks the 3' end of DNA, causing it to bind to Tyr723 on the TopoI enzyme, which allows DNA to unwind and relax; it is eventually then attacked by the 5' end of the DNA and the two ends reunite in a process called religation. Topoisomerase I basically holds the 3' end for a bit while DNA gets replicated.^[43] Topoisomerase I inhibitors can prevent DNA from religating, and induce cytotoxicity (cell death); which is actually a good idea for cancer cells that overexpress Topoisomerase I as they die much faster than regular cells.

Evodiamine has an IC₅₀ value of 6.02uM in MCF-7 breast cancer cells, which express high levels of Topoisomerase I, and only slightly less cytotoxic than Camptothecin which is a research standard drug.^[44] Evodiamine is able to prevent DNA religation in a concentration-dependent manner in a similar manner to Camptothecin, by making a complex with Topoisomerase I and the 3' DNA strand it holds.^[44] Evodiamine has demonstrated cytotoxixity in breast cancer cells elsewhere^[45] and was shown in adriamycin-resistant breast cancer cells to induce cell death both in vitro and in vivo with a potency greater than that of paclitaxel.^[46]

Evodiamine is a dual catalytic topoisomerase I and II inhibitor,^[47] and shows efficacy against some cells that are resistant to the more potent topoisomerase I inhibitor Camptothecin,^[47] and does not seem to induce DNA damage.^[47] A flavanoid from evodiamine also possesses dual inhibitory potential.^[45]

A mechanism by which Evodia Fructs may be anti-cancer; awaits more studies to see its overall clinical relevance, but its dual inhibition is novel and promising at least

Apoptosis, or the event of cell death, is an important biomarker in cancer therapy.

Evodia components have been demonstrated to induce apoptosis in gastric cancer cells (SGC-7901)^[48][49][50] and (N-87),^[10] Breast cancer cells (MCF-7)^[51][45][44] and (NCI/ADR-RES),^[52] Liver cells (HepG2),^[10][53] Leukemia cells (HL-60)^[10] (THP-1)^[51] and (U937),^[54] Lung (H-460)^[10] and (LLC),^[55] Colon (COLO-205),^[56][57] Thyroid (ARO),^[58] Melanoma (A375-S2),^{[59][60][51][61]} and (B16-F10),^[55] Cervical Cancer (HeLa),^[62][51] fibrosarcoma (L929),^[51] and has been demonstrated in vivo to inhibition the Sarcoma-180 tumor model when fed as the decoction Zuo jin wan.^[63][64] More often than not, these effects are attributed to Evodiamine and its metabolites although in some instances flavanoid glycosides are to credit.

When rutaecarpine (active indole in Evodia) is fed to rats, the AUC and C_max values of caffeine are significantly reduced; meaning rutaecarpine can reduce the exposure of caffeine to the body.^[65][66] This also extends to the similar xanthine compound theophylline.^[67] These effects extend to consumption of Evodia itself, and combination decoctions such as Wu Zhu Yu Tang.^[66]

This effect is rather significant; pretreatment of 80mg/kg oral Rutaecarpine daily for 3 days (a high dose, but its bioavailability is unaugmented) in rats reduced the C_max to 31% of the control group, the T_max to 22%, the AUC to 5% of control, and reduced the half-life from 0.73+/-0.07 hours to 0.27+/-0.1 hours.^[65] Similar trends were seen for all metabolites of caffeine (paraxanthine, theophylline, theobromine)

Rutaecarpine is able to induce (increase) activity of various hepatic enzymes such as CYP1A2, CYP2B, and CYP2E1;^[27][68] CYP2E1 induction is seen at 80mg/kg oral ingestion in rats while only 20mg/kg is needed for CYP1A2.^[66] As caffeine is highly metabolized by CYP1A and CYP2E1, their increased activity causes greater metabolism of caffeine in a shorter time frame and thus limits systemic exposure.

Rutaecarpine appears to potently suppress the circulating levels of caffeine via increasing hepatic and intestinal degradation; a highly antagonistic compound

The pair of Evodia fruit and Rhizoma Coptidis (rhizome of Coptis chinensis) is known as Zuo jin wan and is a Traditional Chinese Medicine decoction for acute gastrointestinal distress. It is paired with 6 parts Rhizoma Coptidis to 1 part Evodia Fructus (6:1 ratio). Conversely, reversing the ratio and favoring Evodia in the 6:1 ratio is the basis for Fan zuo jin wan, another Chinese decoction. These formulas are indicated for short-term use only.^[69]

Coptis and Evodia form a dichotomy of cold and hot (Yin and Yang), respectively, where Coptis reportedly induces a cold state and a seeking process for a warm environment while Evodia induces heat and the seeking behavior for a cold environment.^[70] Coptis seems to be able to reduce internal body temperature and oxygen consumption in mice and prolong time spent in a warm environment (as assessed by warm pads) while Evodia increases time in a cold environment, and increases both oxygen consumption and body temperature.^[70] Zuo jin wan is classified as cold whereas Fan zuo jin wan is classified as warm.^[69][70] The only currently known biomarker for this temperature preference is liver ATPase activity.^[71][72]

Coptis Chinensis and Evodia Fructus appear to be mostly antagonist to each other in how they are seen to affect temperature; yet they are used together in decoctions and mixtures (for acute conditions and short-term usage; a 'harmonizing of hot and cold' treatment strategy). The historical reports of Evodia's 'heat' and Coptis's 'cold' properties actually do seem to have some merit, as evidenced by the rat preference tests trying to bioregulate their temperature

When looking at the pharmacokinetics of the combination, Rhizoma Coptidis appears to benefit the pharmacokinetics and circulating levels of some bioactives in Evodia such as dehydroevodiamine^[20] by about 274% of the value of Evodia alone despite the same oral dosage (AUC value).^[73] Evodia seems to either not significantly affect the pharmacokinetics of some Coptidis molecules such as coptisine (acutely)^[20] yet increase general absorption^[74] but can reduce the AUC of various alkaloids (coptisine, palmatine, jateorrhizine) after prolonged ingestion and reduce the AUC and C_max values of the berberine content of Coptidis Rhizoma either chronically^[74] or acutely.^[73] This was hypothesized to be secondary to Evodia Fructus pretreatment enhancing the expression of hepatic UGT1A1, a sulfation enzyme, which conjugated compounds in Coptidis Rhizoma.^[74]

Coptidis chinensis rhizome enhances the bioavailability and circulating amount of co-ingested Evodia fruit, but Evodia fruit hinders the absorption and circulating amount of the active ingredients of Coptidis Chinensis; almost as if the former is sacrificing itself for the latter

Paeoniflorin is the main bioactive found in the herb Paeonia Lactiflora, and its bioavailability and circulating amounts are enhanced by various herbs. Although not to the same degree of Fennel fruit (which increases bioavailability to 226.02% of the level of Paeoniflorin in isolation), consumption of Paeoniflorin alongside Evodia fruit elevates relative absorption to 123.62%.^[75]

Paeonia Lactiflora doesn't do much for Evodia, but Evodia enhances the absorption of Paeonia Lactiflora bioactives