The tradicional medicinal herb III: Andrographi Paniculata

1. Sources and Composition

1.1. Sources

Andrographis Paniculata (of the family Acanthaceae) is a herb with traditional usage for fighting off colds and infection (usually in Chinese medicine[6])[7][8] although antibacterial, antifungal, antiviral, choleretic, hypoglycemic, hypocholesterolemic, and Adaptogen-like effects have been reported as well as being a gastric (stomach) and hepatic (liver) tonic.[9]The usage of Andrographis Paniculata as a liver tonic is seen more in Ayurveda than it is in Traditional Chinese medicine[6] and is used as an anti-neoplastic agent in Ayuredic medicine.[10][6]

It also goes by the names Chiretta (or Chirayetah in Urdu), King of Bitters (as a testament to its bitter properties in Traditional medicine as well as taste), and Kalmegh (Hindi);[8][9] in english it is referred to as Creat[9] and in Traditional Chinese Medicine it is referred to as Chuanxinlian, Yijianxi or Lanhelian.[9] It is a component of a Chinese medication known as Xiang-Qi-Tang alongside Astragalus membranaceus and Cyperus rotundus.[11]

Although all parts of the plant have traditionally been reported at times, the leaves are the most common medicinal part of this plant.[9]

 Has usage in traditional medicine for both being a liver tonic and anti-cancer agent (Ayurvedic medicine) and is most popular for its usage in Traditional Chinese Medicine for reducing the severity and length of cold/flu symptoms

1.2. Composition

Andrographis Paniculata tends to contain in the leaves:

• Andrographolide at (commonly seen as the main bioactive[12][8] at 24.72-43.16mg/g in the flowers[13]but 0.81-1.86% (81-186mg/g) overall[14]) with up to 4% being reported.[12]
• Isoandrographolide, Bisandrographolide A[15] and Neoandrographolide (16.65+/-4.48 mg/g in the cotyledons[13])
• 14-deoxy-11,12-didehydroandrographolide
• 14-deoxyandrographolide at 10.67-24.54mg/g (1-2.5%)[13] with highest content in young leaves[13]
• Andrograpanin[12]
• Skullcapflavone I
• 7-O-Methylwogonin[12]
• Various methoxyflavones[16][17][18][19]
• 3,4-Dicaffeoylquinic acid[12]
• Onysilin[12]
• β-sitosterol, stigmasterol, and ergosterol peroxide[18]

Despite the above quantification of Diterpenoids (Andrographolide and related names), products derived from the herbs tend to have variable quantities (market assessment of products);[20] and one review[12] noting that the Andrographolide content in the leaves can vary from between 0.5% and 6% dry weight.[20][21][22]

 The Andrographolide class of nutrients are the active ingredients, and in the plants themselves they have large variability

2. Molecular Targets

2.1. TRPs

TRPV4 is a calcium channel that, upon activation, increases intracellular calcium and has been implicated in osmoregulation, nociception, regulation of vascular tone, and heat sensation[23] with some possible roles in heat regulation[24] and usually the pharmacological goal is to antagonize the channel.[23]

Bisandrographolide A has been noted to be an agonist of the TRPV4 receptor with an EC50of 790-950nM without having any affinity for TRPV1-3; Andrographolide per se was inactive on any receptor.[15]

 One of the secondary bioactives is a potent and highly selective TRPV4 receptor activator

3. Pharmacology

3.1. Absorption

In an oral bioavailability in rats using 20mg/kg of Andrographolide Paniculata (1mg/kg Andrographolide) it was noted that serum Andrographolide has a bioavailability of 91% at this dose, with ten-fold the dose (10mg/kg Andrographolide) having a bioavailability of 21.4%[25] and one study using 120mg/kg Andrographolide noting a poor absorption rate of 2.67%.[26]

The decreasing bioavailability with higher doses appears to be related to excessive P-Glycoprotein efflux, and incubation with Vermapril can increase its absorption and accumulation into cells.[26]

 Appears to have great absorption at low doses (1mg/kg in rats, equivalent of 0.16mg/kg in humans or about 11mg for a 150lb human) with progressively reduced absorption at higher doses; this may underlie its apparently high toxicity threshold (large safety buffer)

3.2. Serum

In rats fed 20mg/kg Andrographolide Paniculata (1mg/kg Andrographolide) it was noted that Andrographolide had a Cmax of 1.273mcg/mL at a Tmax of 2.41 hours with an AUC of 7.09mcg/h/mL and a half-life of 2.4 hours.[25]

In rats fed 200mg/kg Andrographolide Paniculata (10mg/kg Andrographolide) it was noted that Andrographolide had a Cmax of 3mcg/mL at a Tmax of 1.67 hours with an AUC of 15.07mcg/h/mL and a half-life of 2.9 hours.[25]

In humans using Kan Jang capsules (combination Andrographolide and Acanthapanax senticocus) totalling 17mg Andrographolide, oral consumption resulted in a serum concentration of 141.7+/-20ng/mL with an approximately Tmax of 1.36 hours and a half-life of about 25 minutes, with no detectable Andrographolide detectable in the blood 8 hours after oral administration (some persons having no serum levels after 4 hours).[25] The authors the hypothesized what the steady state value would be following traditional usage (this dose, taken thrice a day) and calculated 660ng/mL; human values obtained in this study were predicted well with rat parameters obtained earlier, suggest they are similar.[25]One other study in healthy male volunteers noted a Tmax at 1.6 hours with a Cmax of 58.62ng/mL following oral ingestion of 200mg isolated Andrographolide.[27]

3.3. Metabolism

Following ingestion of Andrographolide, sulfated metabolites are found in the urine[28] and in particular 14-deoxy-12(R)-sulfoandrographolide[29] whcih appears to be structurally identical to an anti-inflammatory drug marketing in China under the name Lianbizhi.[30]|published=1981 Aug|authors=Meng ZM|journal=Yao Xue Xue Bao][26]

3.4. Distribution

Andrographolide has been noted to bind to bovine serum albumin (BSA) with an association constant of 2.59 and 5.52 binding sites and maximal binding of 79.2%.[25] 64% of Andrographolide is thought to bind to Bovine Serum Albumin at physiological concentrations, and 55% to human serum albumin.[25]

Andrographolide has been noted to distribute into tissues of persons consuming it with variability, with this study noting that it was fairly rapid tissue accumulation after absorption in 25% of the sample (n=4) with accumulation occurring after 1.5-3 hours in the rest of the sample.[25]

3.5. Excretion

It appears that minimal Andrographolide per se is excreted in the urine (rats), with 8.2% of the oral dose beinge excreted in the urine within 72 hours (8.75% overall) and having an elimination rate of 0.128 hours-1 and renal clearance of -0.028 ml/min.[25] The authors suspected with metabolic transformation or fecal excretion accounted for the majority.[25]

3.6. Phase I Enzyme Interactions

An ethanolic extract of andrographis paniculata appears to have mixed type inhibitory potential against CYP2C19 with a IC50 of 91.7μg/mL.[31] The methanolic and hexane extracts were weak inhibitors (IC50 values of 123.3μg/mL and 107.1μg/mL; respectively), the water extract showed no inhibitory potential, and andrographolide itself was very ineffective (IC50 greater than 1426.7μM).[31]

3.7. Phase II Enzyme Interactions

Andrographus paniculata (95% ethanolic extract) appears to exert relatively potent inhibitory effects against various enzymes of drug metabolism including UGT1A1 (IC50 of 5μg/mL), UGT1A3 (1.7μg/mL), UGT1A6 (5.66μg/mL), UGT1A7 (9.88μg/mL), UGT1A8 (2.57μg/mL), UGT1A10 (15.66μg/mL), UGT2B7 (2.82μg/mL) although the inhibitory effects on UGT1B15 were greater than 50μg/mL.[32]

4. Neurology

4.1. Neuroinflammation

0.78mg/kg of Andrographis Paniculata in mice for a week prior to LPS injections (pro-inflammatory stimuli) noted that CXCL2 (aka. MIP2) mRNA was significantly suppressed relative to LPS control; 1.52-3.12mg/kg were less effective, but as effective as the active control of 50mg/kg pyrrolidine dithiocarbamate.[33] This study also noted no significant influence of MIP2 mRNA in the liver of the same mice, suggesting a localized effect.[33] MIP-2 is involved in neutrophil accumulation[34] and when active in the brain have been noted to increase blood-brain barrier permeability to immune cells to facilitate recruitment.[35]

5. Cardiovascular Health

5.1. Cardiac Tissue

Incubation of cardiac muscle with 5-320uM of 14-deoxy-11,12-didehydroandrographolide (DDS) has been noted to cause a concentration-dependent reduction in the rate of atrail beating (heart rate).[36]

5.2. Blood pressure

14-deoxy-11,12-didehydroandrographolide (DDS) appears to cause dose-dependent reductions in blood pressure[37][36] (which has led to it's quantity being restriced in some supplements to avoid possible hypotension[8]). DDS is more effective at reducing blood pressure than andrographolide and neoandrographolide in vivo[37] and appears to have an ED50 value of 3.43mmol/kg (in anaesthetised rats).[36]

The reduction in blood pressure appears to be mediated via adrenoreceptors, and is thought to act as a beta-blocker (antagonist of the beta-adrenoreceptors) as captopril and propanolol both attenuated the effects with the latter nearly abolishing it.[36]

5.3. Platelet

Andrographolide is able to concentration-dependently (25-100uM) cause apoptosis of platelets via a mitochondrial dependent pathway, which was inhibited by the caspase-8 inhibitor z-IETD-fmk (suggesting capsase-8 is critical to Andrographolide-mediated apoptosis).[38]

6. Interactions with Immunology

6.1. Mechanisms

In LPS-activated macrophages, 14-deoxy-14,15-dehydroandrographolide was able to inhibit NF-kB activation with an IC50 of 2μg/mL[18] although up to 10-20μg/mL of the ethyl acetate extract from Andrographis Paniculata is required for such inhibition.[33] At least one study noted that in vitro efficacy of the plant extract (ethyl acetate) is similar to Morus albaand Eucommia ulmodes leaves while being more effective than Astragalus membranaceusand trended to be less effective than Isatis indigotica (all tested in vitro between 2-5μg/mL).[33]

This has been noted elsewhere with the ethyl acetate fraction of Andrographis Paniculatafollowing oral ingestion of 0.78-3.13mg/kg in mice injected with LPS after Andrographis feeding for a week; this study noted that 6.25mg/kg failed to reduce lethality from LPS like lower doses.[33]

 Some compounds may have anti-inflammatory effects, with fairly potent effects in vitro

At 1µM, lymphocyte proliferation and IL-2 secretion is enhanced by Andrographolide (14%), 14-deoxyandrographolide (5%), and 14-deoxy-11,12-didehydroandrographolide (7%) although the dichloromethane and methanolic extracts (which contain these molecules) are more potent, suggesting synergism from other bioactives.[39]

 On immune cells, Andrographolide may possess weak to moderate immunostimulatory properties

6.2. T-Cells

15-30mg Andrographolide daily for up to 6 weeks has been noted to decrease CD4+ T-cell count by 27% in healthy control persons after 6 weeks while increasing CD4+ cell count in persons with HIV by 23.7% at 6 weeks.[40]

6.3. Sickness and Infection

Andrographis Paniculata is a popular Traditional Chinese Medicine for the purposes of reducing cold and flu symptoms. Surprisingly, this herb may not possess any anti-bacterial effects.[41]

In persons (n=152) with pharyngotonsillitis, daily ingestion of 3-6g of Andrographis Paniculata were effective at reducing signs and symptoms of pharyngotonsillitis with the higher dose (6g) being as effective as the active control of Paracetemol (Tylenol).[42] Benefits were seen on day three of treatment, with no further benefit seen when measured on day 7.[42]

In persons with uncomplicated Upper Respiratory Tract Infection (URTI) taking 200mg of an extract from the leaves of Andrographis Paniculata (31.3% Andrographolide) in two divided doses of 100mg daily for 5 days noted that while there were no differences between groups at baseline or at three days that on day 5 the Andrographis group experienced reduced symptoms (such as Expectoration, Headache, Cough, Fever, and Fatigue; Earache was the only unaffected parameter).[8] These seemingly general benefits are also noted with 1200mg of the basic plant extract, where 4 days of supplementation was able to reduce all measured symptoms of the common cold relative to placebo while sore throat, nasal secretion, and earache were significantly reduced after 2 days.[43]

 In regards to Andrographis alone, it appears to be effective in a general manner against respiratory infections

For the purposes of general immunity, Andrographis Paniculata appears to be used in combination with Siberian Ginseng (Eleutherococcus senticoccus), which is either known as SHA-10 (standardization) or 'Kan Jang' tablets; the latter due to the combination also being used in Traditional Chinese Medicine.[1] There appears to be a large amount of human studies originating from Russia in support of this combination (cited from this study,[1]cannot be located online and conducted between 1967-1995).

A pilot study using Kan Jang tablets (SHA-10) with both Andrographis Paniculata (85mg standaridized for 5.25mg Andrographolide plus Deoxyandrographolide) and Siberian Ginseng (9.7mg containing 2% Eleuthroside B and E) with four tablets taken thrice a day (63mg Andrographolides and 116mg Siberian Ginseng daily) noted that the improvement in overall symptoms in persons with URTIs were to a greater degree than placebo over 5 days.[1] These results were later replicated in a Phase III Trial of 180 persons, where Kan Jang capsules at this dose outperformed placebo over the course of 5 days in reducing symptoms from URTI.[1] Similar effects have been replicated elsewhere in a sample of 185 persons using similar dosing, although this latter study noted a larger effect size (while placebo experienced a 23% reduction in symptoms, Kan Jang experienced an 86% reduction) with most significant improvement in throat symptoms (dry and soreness), headache, malaise, sneezing, and runny nose with some efficacy on cough.[2]

One comparative study pitting Kan Jang capsules against Echinacea purpurea using either for a 10 day period in children with colds noted that Kan Jang was significantly more effective than Echinacea.[4]

One study using Kan Jang capsules with additional Schisandra chinensis and Licorice(48mg Andrographolide daily) for one month has noted significantly less symptoms (relative to placebo) in persons with Familial Mediterranean Fever.[5]

 Kan Jang tablets (combination therapy of Andrographis paniculata and Eleutherococcus senticoccus) appears to be effective in reducing the signs and symptoms of upper respiratory tract infections when taken at the onset of sickness. At least one study suggests it is more potent than Echinacea purpuera

6.4. Virology

In 13 HIV-positive persons given Andrographolide (PN355) at 5mg thrice a day (totalling 15mg) for 3 weeks in escalating dose (10mg for another 3 weeks with the trial ending at 6 weeks) noted a high degree of side-effects associated with treatment (no placebo for comparison) and noted an increase in hepatic enzymes (ALT and AST) associated with treatment and normalized 3 weeks after cessation.[40] There was no influence on HIV RNA content.[40]

6.5. Arthritis

Supplementation of Andrographis extract (30% Andrographolide) at three daily doses of 100mg (300mg daily) for 14 weeks in persons with Rheumatoid arthritis failed to significantly reduce joint pain although both the count and degree of tender and swollen joints was significantly reduced with Andrographis intervention.[44] This study also noted reduced serum IgA and complement component C4.[44]

 At least one study noted that Andrographis may reduce swelling (but not significantly reduce pain) associated with Rheumatoid Arthritis

7. Interactions with Hormones

7.1. Testosterone

One study using 20-1000mg/kg of Andrographis Paniculata (10.9% Andrographolide) daily for 60 days has failed to find any significant influence on circulating testosterone levels.[45]

8. Interactions with Organ Systems

8.1. Liver

Mechanistically, in HepG2 cells treated with CCL4 both Andrographolide and Andrographis paniculata extract (2.7% Andrographolide) exert cytoprotection when coincubated with maximal protective effects at 30µmol/mL Andrographolide which preserved 84.3% of cell viability (higher concentrations not tested);[46] this was said to be due to its antioxidative capacity (IC50 of 3.2µg/mL (9.2µmol/mL) in DPPH assay with a maximum potency at 15µmol/mL, more potent than Vitamin C which had an IC40 of 4.2µg/mL and reached maximal potency at 40µmol/mL).[46] Protective effects against CCL4 hepatotoxicity have been noted in vivo when mice were pretreated for 8 days with 50-100mg/kg Andrographolide, where only the higher dose of 100mg/kg was associated with a reduction of ALT and AST enzyme secretion with reduced MDA (indicative of lipid peroxidation) and increased glutathione; 100mg/kg Andrographolide was equally effective as the active control of 100mg/kg Silymarins (from Milk Thistle).[47]

Andrographolide has also shown protective effects in H4IIEC3/G(-) liver cells induced with acetominophen toxicity, where 10µM of Andrographolide preserved 55% of cell viability and outperformed 10µM of Kutkin from Picrorhiza kurroa (46%) and Silymarin (24%).[48]

 Appears to have general hepatoprotective properties against pro-oxidative toxins, with one study suggesting it is dose-for-dose as effective as Silymarins from Milk Thistle (although Andrographus paniculata supplements tend to be dosed much lower inherently)

One pilot study in persons with HIV (small group of persons without HIV) noted that treatment of 15-30mg Andrographolide in three divided doses was associated with an increase in serum liver enzymes (AST and ALT) in both groups, with the HIV group normalizing liver enzymes 3 weeks after supplement cessation.[40]

8.2. Lung

Andrographolide pretreatment to mice 2 hours prior to exposure to smoke fumes has been noted to be associated with less fluid accumulation and changes in biomarkers thought to be due to antioxidative effects (reduced lipid peroxidation and increase antioxidative enzymes); this was thought to be downstream of Nrf2 activation, which was noted in vitro(BEAS-2B cells) to be induced by Andrographolide treatment.[49]

8.3. Kidneys

In a series of case studies of persons who underwent Extracorporeal Shock Wave Lithotripsy (ESWL; a kidney stone treatment[50]) given two doses of 1,000mg Andrographis Paniculata daily for 5 days following treatment noted reductions in pre and post-ESWL pyuria, hematuria and proteinuria of similar potency to the active controls of Norfloxacin (200mg) or Cotrimoxazole.[51] These results suggest that Andrographis Paniculata has renoprotective properties.

8.4. Intestines

In animals with colitis given a water extract of Andrographis Paniculata, inhibition of CD4+ T-cell differentiation into TH1/TH17 cells was said to underlie the observed protective effects of supplementation where the signs and symptoms of colitis were abolished in mice recieving both supplementation and methyl celluose (which induced the colitis);[52] this study is duplicated in Medline.[53]

An extract known as HMPL-004 at 1200mg (thrice daily dosing of 400mg) in persons with Ulcerative Colitis daily for 54 days (on average) noted that the rate of clinical remission was 21% (clinical signs) or 28% (colonoscopy) with the rate of response being 76% and 82% (clinical and colonoscopy, respectively); the active control of 4500mg Slow-Release Mesalazine achieved similar rates of remission (16% and 24%, respectively) and response (74% and 71%).[54] Another trial using 1200mg and 1800mg in adults with mild-to-moderate colitis noted a dose-dependent increase in 'response to treatment' (reduction of symptoms associated with colitis) which trended towards significance at 1200mg but was significant at 1800mg; no significant differences were noted in side-effects relative to placebo.[55]

 Appears to show promise in Ulcerative Colitis, with one double-blind and one comparative study coming back positive currently

8.5. Testes

Interestingly, one study on Eurycoma Longifolia Jack showed the efficacy of said supplement in a rat model of "Andrographis Paniculata induced infertile rats".[56] This may have originated from a study in male albino rats where 20mg powder for 60 days induced infertility and a cessation of spermatogenesis[57] and another study noting that 25-50mg/kg isolated Andrographolide for 48 days inducted testicular toxicity and altered spermatozoa.[58]

In opposition to the above, doses of 1000mg/kg Andrographis Paniculata (6.1% Andrographolide) for 60 days in male rats has failed to exert appreciable testicular toxicity[22] and these lack of effects persist when looking at fertility of male rats over 65 days, where the lack of testicular toxicity was replicated.[45] It should be noted that the latter two studies appear to have better methodology than the former two (involving electron microscope analysis of testicular histology and measuring serum levels of biomarkers).[45]

Additionally, a Phase I trial in healthy humans using Kan Jang capsules (the anti-cold Chinese Medicine where Andrographis is paired with Siberian Ginseng) at thrice the recommended dose for 10 days failed to find any toxic effects on sperm cells.[3]

 Has been implicated in inducing testicular toxicity, although this is a contested issue and the one human study using feasible oral doses has failed to find any signs of testicular toxicity

9. Interactions with Cancer

9.1. Immunological Interactions

In lymphocytes (obtained from volunteer) treated with 1μM Andrographolide, an increase in CD3 (61–91% relative to control lymphocytes), CD4+ (40–61%), CD8+ (23–31%), and CD56 (2–3%) marker expression was noted and associated with enhanced lymphocyte-mediated toxicity on K562 cancer cells; this was also noted to occur at 0.1μM.[59]

 May enhance lymphocyte-mediated toxicity towards some cancer cells

Treatment of cancer cells with Andrographolide was associated with upregulation of Death-Receptor 4 (DR4) which enhanced TRAIL-mediated apoptosis; this was downstream of p53 induction and abolished with coincubation of N-Acetylcysteine (and thus was mediated by pro-oxidative mechanisms).[60] TRAIL is a selectively tumor cytotoxic agent from the immune system[61] which is a therapeutic target in cancer research.[6][62]

 Appears to enhance TRAIL-mediated cytotoxicity in cancer cells via pro-oxidative (possibly hormetic) means

Andrographolide is able to enhance cytotoxicity of reference cytotoxic drugs (5-fluorouracil, adriamycin, and cisplatin) when coincubated in multi-drug resistant colorectal cells[63] where it synergistically enhances the cytotoxicity of other agents.[64]

 May confer aid in multi-drug resistant cells

9.2. Colon Cancer

In human colorectal carcinoma cells (Lovo) noted that Andrographolide was accumulated up to 0.61+/-0.07μM/mg (12 hours) in cells and displayed a IC50 on cell growth of 8.6μM at 24 hours, with 48 hours incubation of 10-30μM causing near inhibition of proliferation.[65]One study assessing a wide variety of tumor cell lines noted that while the IC50 ranged from 5-15μM that one colon cell line (COLO205) was more sensitive with an IC50 of less than 1μM, although inhibitory effects were noted on all five tested colon cancer cell lines (SW620, HCT116, HT29, KM12, and COLO205)[59] and elsewhere noted to be 11μM on SW620.[39] Andrographolide is not the only molecule in Andrographis Paniculata to have cytotoxic effects on colon cancer cells, but the methylated flavones appear to have higher IC50 values (less potency) and most other diterpenes related to Andrographolide are about 3-fold less effective, although this study noted that 14-deoxy-14,15-didehydroandrographolide was similarly effective on COLO205 cells;[66] this potency of Andrographolide relative to other diterpenes has been noted in Leukeia cells as well.[67]

 Andrographolide appears to have potent anti-proliferative effects on a wide variety of colon cancer cells; the cytotoxic effects (ability to kill cells, in this context we mean tumor cells) of Andrographolide are currently not as well researched and some sources hint at it being quite less potent or unremarkable

Andrographolide has been noted to accumulate cells in the G1 phase of the cell cycle (43.28% to 56.05% at 10μM) with a decrease in S fraction in a concentration and time dependent manner; this was associated with a downregulation of Cyclin D1 (no significant influence on Cyclin D3 or E although both Cdk2 and Cdk4 were reduced in protein content) all thought to be downstream of p53 induction of protein content and phosphorylation.[65]

An increase in the Rb/E2F complex has been noted with Andrographolide,[65] and secondary to more of this complex less free E2F is released; E2F being a factor in cell proliferation and its complexation reducing proliferation.[68] One other study has noted anti-invasive effects of Androgapholide on colon cancer cells (CT26 and HT29) although attributed this observation to inhibition of MMP2 (and inhibition of Akt),[69] a protein that promotes invasiveness of tumor cells. Studies in lung cancer cells suggest that MMP2 inhibition may also be downstream of HLJ1 upregulation.[70]

 The anti-proliferative effects appear to be mediated by modulation of Cyclin proteins, which appear to be downstream of p53; other evidence (not on colorectal cells) hints that p53 is downstream of prooxidative and hormetic effects

9.3. Breast

Andrographolide has been noted to induce apoptosis in breast cancer cells at 0.35-1.4mM in TD-47 breast cancer cells in a concentration and time dependent manner, with apoptosis at 24 hours ranging from 28.55-65.65% and at 72 hours ranging from 76.48-99.72%.[71]This apoptosis was associated with DNA fragmentation associated with an increase in p53 content,[71] and has been noted to be a pathway independent of the mitochondria.[72]

The related structure 14-deoxy-11,12-didehydroandrographolide has been shown to induce both apoptosis and autophagic structures (visually assessed) in TD47 breast cancer cells at 1.5μg/ml[73] (the IC50 found here[74]) and these changes were associated with 2-fold modifications of genes involved in cell cycles (19), cell growth and proliferation (42), tumor suppression (40), apoptosis-related mechanisms (18), and vesicle formation and transport as well as protein degradation (30); all named within the citation.[73]

An increase in G1 phase cells with concomitant decrease in S phase (as well as G2/M) has been noted in MCF-7 cells (treated with 5μM Andrographolide) at 24 hours with an increase in sub-G1 cell content at 48 hours.[59] Andrographolide was noted to induce p27 content and subsequent downregulation of Cdk4 (at 25μM).[59]

 There is apparent cytotoxicity associated with Andrographolide in brast cancer cells, but at a very high concentration that may not be practically relevant

Growth inhibitory effects have been noted on four breast cancer cell lines (MDA-MB-453, MCF7, MCF7/ADR and T47D) with an IC50 value ranging from 5-15μM[59] and another study using NCI/ADR-RES noting an IC50 of 15μM with Andrographolide and 30μM with 14-Deoxy-11,12-didehydroandrographolide.[39]

 There appears to be standard anti-proliferative effects at fairly low concentrations of Andrographolide

Anti-tumor activities of Andrographolide have been confirmed in vivo in mice transplanted with tumors, and was associated with PI3K/Akt and suppression of angiogenetic factors such as VEGF and Osteopontin.[72]

9.4. Leukemia

In studies done on leukemic cells, it is noted that while Andrographolide has cytotoxic activity that both 14-deoxy-11,12-didehydroandrographolide and neoandrographide have failed to exert any cytotoxic activity[67] and that Andrographolide-mediated cytotoxicity in HL-60 leukemic cells occurs via mitochondria-dependent mechanisms.[75]

Subsequently, Andrographolide has shown anti-proliferative properties in two Leukemic cell lines (CCRF-CEM, RPMI8226) with an IC50 between 5-15μM[59] and in HL-60 leukemic cells a 27% increase in G0/G1 cell count at the expensive of S and G2/M at 12μg/mL over 36 hours.[75]

One study noting the IC50 of proliferation on Jurkat Lymphoma cells relative to healthy PMBCs noted that two dehydroandrgrapholides and Isoandrographolide had IC50 values of 50-100μM while the IC50 in healthy cells was either 150μM or 'greater than 200μM'.[66]

 Andrographolide has shown to have anti-proliferative effects in Leukemic cells at regular concentrations, with apoptosis being induced at higher concentrations

At least one study has noted that Andrographolide was able to influence differnetiaiton of myeloid leukemia (M1) cells of mice into phagocytes,[76] which is not a common mechanism among plant-derived nutraceuticals.[6]

9.5. Melanoma

Isolated Andrographolide has shown anti-proliferative effects in isolated Melanoma cells (A431, M14, UACC62) with an IC50 between 5-15μM[59] with another study using M14 cells noting an IC50 of 11μM.[39]

In mice planted with B16 melanoma tumors, oral ingestion of Andrographolide at 100-200mg/kg oral dose (human equivalent of 8-16mg/kg) was associated with a suppression of tumor growth at 30-36% (100mg/kg) and 39-52% (200mg/kg) over the course of 10 days.[59]

9.6. Ovarian

Isolated Andrographolide has been noted with inhibiting proliferation of four ovarian cancer cells (ES2, SKOV3, OVCAR8, and PA-1) with an IC50 between 5-15μM[59] with another study noting an IC50 (SKOV3) of 18μM.[39]

9.7. Prostate

Isolated Andrographolide has been noted with inhibiting proliferation of two prostate cancer cells (DU145 and PC3) with an IC50 between 5-15μM[59] with other studies noting growth inhibition (IC50) of 12μM (PC3)[39] and 70-150μM (PC3).[66]

9.8. CNS

Isolated Andrographolide has been noted with inhibiting proliferation of three CNS cancer cells (U251, SF268, and SNB19) with an IC50 between 5-15μM[59] with another study replicating these growth inhibitory effects on U251 with an IC50 of 10μM.[39]

In glioblastoma cells (U251 and U87) Andrographolide has been noted to inhibit proliferation in a concentration and time dependent manner between 10-100μM (no cytotoxicity noted) associated with G2/M arrest, increasing the percentage of cells in G2/M phase from 15.73% to 35.15% (123% increase in U251) and 16.99% to 33.61% (98% increase in U87) at 70μM.[77] This was associated with downregulation of Cdc25C and Cdk1 as well as slight inhibition of PI3K/Akt, although coincubation with a more potent PI3K/Akt inhibitor enhanced the anti-proliferative effects greatly.[77]

 Anti-proliferative effects on glioblastoma cells appear to be synergistic with the mechanism of PI3K/Akt inhibition

9.9. Lung

Isolated Andrographolide has been noted with inhibiting proliferation of six tested lung cancer cells (A549, NCI-H23, HOP62, MES-SA, H522, and MES-SA-DX5) with an IC50between 5-15μM[59] although another study using H522 noted an IC50 of 16μM.[39]Andrographolide has also been noted to enhance HLJ1 promoter activity secondary to JunB activation, which then caused suppressed activity of MMP2.[70]

9.10. Renal

Isolated Andrographolide has been noted with inhibiting proliferation of two renal cancer cells (ACHN and A498) with an IC50 between 5-15μM[59] although another study using A498 noted an IC50 on growth of 28μM.[39]

10. Safety and Toxicity

10.1. General

In interventions with Andrographis Paniculata, side-effects do not appear to be significantly different than placebo[8][2][25][1] even in one instance where side-effects reached 20% of the study group (same as placebo, suggesting another factor was at play);[42] most reported side-effects (regardless of being different or similar as placebo) generally being mild and infrequent.[7]

One study in HIV positive persons has noted that, with isolated Andrographolide, some symptoms similar to an allergic reaction were reported (from two subjects, no placebo group in this study)[40] while according to manufacturers (data retrived here[7]) there have been 5 reported cases of allergic reactions associated with Andrographis Paniculatasupplementation.

 In general, human intervention data appears to show that commonly used doses of Andrographis Paniculata are safe; very few instances of allergic reactions or something sharing the symptoms has been reported

In rats, the LD50 appears to be greater than 17g/kg.