Andrographis paniculata (Burm. f.) Wall. ex Nees

200 Thai Bahts/kg

Cultivated, organic

The plant is indigenous all parts of Thailand. The areas under cultivation are in the western part provinces including Raatchaburi, Kanchanaburi; the middle part provinces including Nakornpathom, Supaanburi; the northern part provinces including Kampaengpet, Pitsanulok, Lampang; the eastern part provinces including Chantaburi; the northeastern provinces including Ubonraatchathani.

Crude, semi-processed and processed

Thai Food and Drug Administration
Web: http://www.fda.moph.go.th/eng/index.stm

Agriculture, Ministry of Agriculture
Web: http://www.doa.go.th/

Royal Forest Department of Thailand 
Web: http://www.forest.go.th/

Thai-China Flavours & Fragrances Industry Co. Ltd. 
(for details see Trade points above)

It was traditionally used as antidiarrhaeal and for treatment of abscesses.

Antibacterial activity: There are many reports of antibacterial activities which some claimed to be broad activities against infections in many parts of the body including upper respiratory tract and gastrointestinal (GI) tract while others reported in controversial way such as no activity or very mild activity. Effective dosages of A. paniculata are very high dosages and there is no dose-response relationship (Ketpanyapong, 1999; Thamlikitkul and Pruksachatvuthi, 1990; Leelarasamee et al., 1990; Sindermsuk, 1993). Extract of A. paniculata can not stop growth of many microorganisms including viruses Herpes Simplex Virus type 2 (HSV-type 2) and Influenza virus; bacteria E. coli ATCC 25922 and S. aureus ATCC 25929; and fungi Candida albican, the Dermatophytes i.e. Epidermophyton floccosum, Microsporum gypseum and Trichophyton mentagrophytes (Pearnpijit, 1993). However, extract of A. paniculata has antibacterial activity against Porphyromonas gingivalis, the prominent oral cavity bacteria causing periodontitis which may apply to use in a gel form for treatment of gingivitis (Komwatchara, 1996; Rassameemasmaung, 1996). In addition, crude extract of A. paniculata was reported to stop adhesion of Streptococcus mutans, the oral cavity bacteria causing teeth decay (Lomsong, 2004). Formulation of A. paniculata in gel form developed as 50% ethanolic extract in glycerol monosterate and soy bean oil was found to poss antibacterial activity against S. aureus and S. epidermidis. However, antibacterial activity declines after preparation. Storage period after freshly prepared of A. paniculata gel is 32 days at 25° C.

Anti-HIV activity: In vitro study of anti-HIV activity by ehydroandrographolide succinic acid monoester, an active constituent of A. paniculata showed positive result at MIC (minimal inhibitory concentration) at 2 µg/ml (Chang et al., 1991; Suttajit et al., 1998).

Antimalarial activity: Some xanthones e.g. 1,2-dihydroxy-6,8-dimethoxy-xanthone from roots of A. paniculata have anti-malarial activity. It showed IC 50 at 4 µg/ml against Plasmodium falciparum (Rajagopal et al., 2003).

Antipyretic and anti-inflammatory activities: Medicinal Plant Research Institute, Ministry of Public Health, Thailand found antipyretic activity of A. paniculata in rabbits and anti-inflammatory activity in rats using method for testing carrageenan-induced hind paw edema (Anonymous, 1990). Ethanolic extract of aerial parts of A. paniculata at 500 mg/kgBW posses equal antipyretic action as aspirin at 200 mg/kgBW when given orally to rats (Vedavathy and Rao, 1991). Dried powder, ethanolic and water extract of A. paniculata has anti-inflammatory property with testing method for carrageenan-induced hind paw edema when given orally to rats and inhibition of white blood cells infiltration and granuloma development were reported by actions of these extracts. Mechanism of anti-inflammatory actions of A. paniculata are related to i) inhibition of nitric oxide (NO) production from inflammatory macrophages by neoandrographolide (Batkhuu et al. 2002) ii) inhibition of NO production by decreasing expression of inducible nitric oxide synthase (iNOS) in macrophases by andrographolide (Chiou et al., 1998; 2000) and iii) inhibition of neutrophil adhesion and transmigration and prevention of reactive oxygen species (Shen et al., 2000; 2002).

Effects on immunity: Many studies on effects of A. paniculata on immunity have been studied by many investigators. Ethanolic extract of A. paniculata and andrographolide, a known compound separated from this plant, were reported to stimulate both specific immunity i.e. antibody response and delayed type hypersensitivity reaction and non-specific immunity by stimulating movement and phagocytosis of macrophages and multiplying of lymphocytes. From this study, ethanolic extract was found to have more potent immune stimulating activity than andrographolide (Puri et al., 1993). Andrographolide and its derivatives; 14-deoxyandrographolide and 14-deoxy-11,12-didehydrographolide found in dichloromethane fraction of methanolic extract of A. paniculata can stimulate division of human peripheral blood lymphocytes (HPBLs) which leading to interleukin-2 production (Kumar et al., 2004). In vitro comparative study on division of human peripheral blood lymphocyte (PBL) was made between andrographolide and Kan Jang, a combined drug composed of the fixed amount of extract of SHA 10 of A. paniculata and extract of SHE-3 of Eleutherococcus senticosus. The results showed that both andrographolide and Kan Jang inhibit cell division of PBL when given single compound each time but increased PBL division via phytohemagglutinin (PHA) activation was observed after coadministered with andrographolide and Kan Jang. Furthermore, stimulating effects of immunity by both compounds have been studied. It has been reported that both andrographolide and Kan Jang stimulate production of interferon-gamma, neopterin, beta-2-microglobulin in blood cell culture and induce production of interferon-alpha and beta-2-microglobulin in whole blood cell culture. In general, Kan Jang was found to be more effective than andrographolide at the same amount (Panossian et al., 2002).

However, immunosuppression of andrographolide was also reported via down- modulation of both humoral immunity (use antibodies) and cell-mediated immunity (use lymphocytes esp. T-lymphocutes). Andrographolide showed many immunosuppressive activities by various actions as following i) interfere T-cell proliferation and cytokine secretion responsible to allogenic stimulation ii) block T-cell stimulation by dendritic cells (DCs) when incubate DCs with andrographolide at antigen pulse stage iii) interfere maturation of DCs and antigen presentation of DCs to T-cells iv) decrease antibody response to thymus-dependent antigen and delayed-type hypersensitivity in mice and v) reduction of experimental autoimmune encephalomyelitis (EAE) symptoms in mice by inhibition of T-cells (Iruretagoyena et al., 2005).

Hepatoprotective effect: Hepatoprotective activity of A. paniculata has been studied both in vitro and in vivo (Choundhury and Poddar, 1984; Handa and Sharma, 1990; Shukla et al., 1992; Udomsksom, 1993; Visen et al., 1993; Kapil et al., 1993; Trivedi and Rawal, 1998; 2000; 2001). It was reported that ethanolic and water extract of leaves of A. paniculata and pure andrographolide can prevent hepatotoxicity caused by toxic chemicals such as carbontetrachloride (Choundhury and Poddar, 1984; Handa and Sharma, 1990; Shukla et al., 1992; Udomsksom, 1993), galactosamine (Handa and Sharma, 1990), paracetamol (Handa and Sharma, 1990; Shukla et al., 1992;  Visen et al., 1993), ethanol and hexachlorocyclohexane (BHC) (Trivedi and Rawal, 1998; 2000; 2001). Andrographolide was reported to have higher hepatoprotective activity than silymarin, a standard drug from milk thistle tree (Silybum marianum) commonly used in treatment of paracetamol overdose and decreasing bile composition and volume (Visen et al., 1993). In study of antidote property of A. paniculata against hepatotoxicity caused by carbontetrachloride in mice, it showed that andrographolide has the similar efficacy as silymarin in reduction of degradation products derived from lipid peroxidation and reduction of glutamate pyruvate transaminase and alkaline phosphatase in serum of experimental animals (Kapil et al., 1993).  

Anticancer activity: Anticancer activities of A. paniculata have been studied in both in vitro and in vivo. Compounds in dichloromethane fraction of methanol extract of A. paniculata can stop growth of intestinal cancer cell HT-29. Andrographolide showed toxic effect to many cancer cells (Kumar et al., 2004). It can inhibit division of many tumor cell lines resulting in G0/G1 stage caused by induction of cell-cycle inhibitory protein p27 and reduction of cyclin-dependent kinase 4 (CDK4). Furthermore, andrographolide was found to increase synthesis of tumor necrosis factor-alpha (TNF-α) and CD marker expression leading to higher cytotoxic activity of lymphocytes to cancer cells. In addition, anticancer activity was shown positive result in B16FO melanoma syngenic and HT-29 xenograft models (Rajagopal et al., 2003).

Cardiovascular effects: Water extract of A. paniculata was found to prevent myocardial infarction after I hr infused intravenously in thrombus-induced dogs. In comparison with a control group, the treated group was found increased PGI2 production, inhibited thromboxane A2 synthesis, increased cAMP in platelets, decreased ischemic area and decreased height of ST segment (Zhao and Fang, 1990). In addition, flavone from extract of roots of A. paniculata showed prophylaxis activity against myocardial infarction derived from thrombus in dogs by prevention of platelet aggregation (Zhao and Fang, 1991). Extract of A. paniculata decreases incidence and severity of atherosclerotic iliac artery stenosis from deendothelialization and high cholesterol intake and helps prevention and reduction of restenosis after angioplasty in rabbits which showed much better results (>70%) than treated with fish oil, which contains high amount of omega 3 polyunsaturated fatty acids. The mechanism of action of this event was also studied (Wang and Zhao, 1993; 1994). Extract of A. paniculata decreases calcium overloading in myocardial ischemic reperfusion injury. The extract can reduce Ca²⁺ level in ischemic region of cardiac muscle cells and reduces increasing Na⁺ and malondialdehyde and prevent reduction of Ca²⁺-ATPase (Kumar et al., 2004). When applied a partial purified compounds of the crude extract known as API0134 via i.v. route in a dog, an evidence of prophylaxis against an increase in left ventricular end diastolic pressure and regulation of cardiac output to normal level, reduction of ischemic ECG and prevention of malignant arrhythmia were observed. In addition, the infarct size was smaller than a control group after reperfusion (Guo et al., 1996). Anti-hypertensive activity was also studied and showed positive results. After intraperitoneal perfusion of water extract of A. paniculata by using osmotic pump, decrease in systolic blood pressure in spontaneously hypertensive rats (HSR) was observed in rats in addition to reduction of plasma angiotensin converting enzyme (ACE) activity and lipid peroxidation in kidneys (Zhang and Tan, 1996). The mechanism of antihypertensive actions of A. paniculata was also studied. The partial purified fraction using n-butanol as extracting solvent from crude water extract was found to have antihypertensive action in anesthetized rats via alpha-adrenoceptor, autonomic ganglion and histaminergic receptors (Zhang and Tan, 1997).

Antioxidative effect: Extract of A. paniculata posses antioxidative property (Chanwitheesuk et al., 2002; Singh et al., 2001; Zhang and Tan, 2000) and it was reported that neoandrographolide is an antioxidant of superoxide free radicals (Kamdem et al., 2002).

Hypoglycemic effect: Ethanolic extract of A. paniculata showed hypoglycemic effect in diabetic rats induced with streptozotocin (STZ) (Zhang and Tan, 2000). Water extract of A. paniculata at dosage of 10 mg/kgBW can decrease blood glucose level after induction of hyperglycemia with glucose orally at 2 mg/kgBW but can not reduce blood sugar activated by adrenaline. However, prolong administration up to 6 months showed no hypoglycemic activity (Husen et al., 2004). Water extract of A. paniculata at 50 mg/kgBW showed 52.90% reduction of blood glucose in diabetic rats treated with STZ while 61.81% reduction of blood glucose was found in diabetic rats treated with freeze-dried form at dosage 6.25 mg/kgBW. Determination of active constituents in A. paniculata was studied and found that andrographolide was one of hypoglycemic agents. After orally given of andrographolide into STZ-induced diabetic rats, it showed hypoglycemic effect in dose-response manner. In addition, andrographolide at 1.5 mg/kgBW can reduce elevated blood sugar level after infusion of glucose intravenously. The mechanism of hypoglycemic effect might be due to glucose uptake capability of the muscle (Yu et al., 2003).

Antiplatelet activity: Andrographolide posses antiplatelet activity by stimulating platelet-activating factor (PAF) in dose-response manner. However, it has no effect on synthesis of eiosanoids or PAF of human polymorphonuclear leukocyte (Amroyan et al., 1999).

Analysis of the whole plant gave the following lactones (dry basis): andrographolide,0.6%; 14-deoxy-11-oxoandrographolide (C20 H28 O5), 0.12%;14-deoxy -11, 12-ldehydroandrographohde (C20,H30O4), 0.06%; 14- deoxyandrographolide (C200H30O4), 0.02% and a non-bitter constituent, neoandrographolide (C26 H40 O8), 0.005%.

The leaves contain andrographolide (yield, 1%). From the petroleum ether extract of the leaves from Bangladesh, the following compounds have been isolated: a, b -unsaturated lactone, homo-andrographolide (C22 H32 O3). andrographosterol (C23 H38 O), andrographane (C40 H82), andrographone (C32H64 0 ), a wax, and two esters containing, hydroxyl groups.

The roots gave apigenin-7,4'-di-O-methyl ether, andrographolide and a new natural flavone, 5-hydroxy-7,8,2',3'-tetramethoxyflavone (C19 H18 07); yield, 0.006%). They also contain a monohydroxy-trimethylflavone, andrographin (C18H1606) and a dihydroxy-di-methoxyflavone, panicolin (C17H14O6 ). The presence of a - sitosterol is also reported.

Chemical markers: Andrographolide, a diterpene lactone is the major constituents of the drug and its minimum quantity is prescribed as 0.5-0.9% in the appropriate drug. Another non-bitter constituent, neoandrographolide is also used as a marker.

The quality control in Thai Herbal Pharmacopoeia Vol.I

Identification 
A. To about 1 g of the sample, in powder, add 20 ml of ethanol, boil in water-bath and filter. To the filtrate, add 300 mg of decolorizing charcoal, stirand filter (solution A). To 1 ml of solution A, add 2 drops of 2% w/v solution of 3,5-dinitrobenzoic acid in methanol and 2 drops of a 5.7% w/v solution of potassium hydroxide in methanol: a purplish red colour develops.

B. To 1 ml of solution A, add several drops of ethanolic potassium hydroxide TS unit it shows a red colour. Set aside for 10 to 15 min the colour is changed to yellow.

C. Carry out the test as describes in the “Thin-layer Chromotography” (Appendix 3.1), using silica gel GF254 as the coating substance and a mixture of 85 volumes of chloroform and 15 volumes of absolute ethanol as the mobile phase but allowing the solvent front to ascend 15 cm above the line of application. Apply separately to the plate, 5 µl of each of the following solutions. Prepare solution (A) by boiling 1 g of the sample, in powder, with 20 ml of ethanol on a water-bath for 5 min, adding 300 mg of decolourizing charcal, stirring, and filtering. Evapurate the filtrate under reduced pressure until dryness, and dissolve 2 mg of andrographolide in 1 ml of ethanol. After removal of the plate from the chromatographic chamber, allow it to dry in air, and examine under ultraviolet light (254 nm), marking the quenching spots. The chromatogram obtained with solution (A) shows a quenching spot (hRf values 52 to 56), correspounding to the andrographolide spots from solution (B) and other four spots of different hRf values (Table 1); see also Fig. III (Appendix 3.1H). Spray the plate with a 2% w/v solution of 3,5-dinitrobenzoic acid in methanol and then with an excess of a 5.7% w/v solution of potassium hydroxyide in methanol; the spot due to andrographolide is dark violet spots are also observed.

Quantitative parameters

• Foreign matter not more than 2% w/w of stems and other foreign matter (Appendix 7.2).
• Acid-insoluble ash not more than 2% w/w (Appendix 7.6).
• Ethanol (85%)-soluble extractive not less than 13% w/w (Appendix 7.12).
• Water-soluble extractive not less than 18% w/w (Appendix 7.12).
• Loss on drying not more than 11% w/w after drying at 105° C to constant weight (Appendix 4.15).
• Total lactones content not less than 6% w/w of total lactones, calculated as andrographolide, when determined by the folling method.

Assay

Basic lead acetate solution: Triturate 14 g of lead (II) oxide with 10 ml of water, add 10 ml of water and transfer into a 100 ml volumetric flask. Add a solution prepared by dissolving 22 g of lead (II) acetate in 70 ml of water, shake vigorously for 5 min, set aside for 1 week, filter, and add sufficient previously boiled water to produce 100 ml.

Procedure: Place about 1 g of Andrographis Herb in No. 180 powder, accurately weiged, in a 100 ml round-bottemed flask, add 50 ml of ethanol (85%), reflux in a water-bath for 2 h, and filter. Wash the marc with sufficient amount of ethanol (85%) until the last washing is almost colourless. Combine the washings and the filtrate and allow to cool. Add 1 ml of Basic lead acetate solution, set aside for 15 min, filter, and wash the precipitate with ethanol until the last washing is no longer green. Combine the washings and the filtrate, add dropwise with swirling 1 ml of a 25% w/v solution of sodium sulfate and mix well. Set aside for 1 h, add 500 mg of decolourizing charcoal, and reflux in water-bath for 10 min. Filter through the Büchner funnel containing 500 mg of decolourizing charcoal and wash with three 2 ml portions of hot ethanol. Combine the washings and the filtrate, add 20 ml of distilled water, allow to cool, and neutralize with 0.1 M sodium hydroxide, using phenolphthalein TS as indicator. Add 5.0 ml of 0.1 M sodium hydroxide VS, reflux in a water-bath for 30 minutes, allow to cool, and titrate with 0.05 M hydrochloric acid VS. Perform a blank determination (Residual Titrations, Appendix 6.17). Each ml of 0.1 M sodium hydroxide VS is equivalent to 35.05 mg of andrographolide, C20H30O5.

HPLC conditions

• Column: Inertsil ODS-3, 5 µm, 4.6 mmI.D. x 15 cm
• Mobile phase: Methanol/water (60:40, v/v)
• Flow-rate: 1 ml/min
• Detector: 254 nm
• Injection volume: 20 µl

Acute toxicity test of 50% alcoholic extract of Andrographis paniculata Wall. Ex Nees. Revealed no evidence of toxicity in mice when given 15 g/kg body weight. LD50 of the extract administered per os and subcutaneous are more than 15 g/kg and intraperitoneal is 14.98 g/kg. Subchronic toxicity test of A. paniculata powder was evaluated for 6 months in 96 wistar rats divided into four groups consisting of control, 0.12, 1.2 and 2.4 g/kg/day equivalent to 1, 10 and 20 times of human therapeutic dose (6 g/day/man 50kg). Toxicity effect of A. paniculata was determined by growth rate (body weight), food consumption, clinical sign, hematological, serum biochemical values and histopathological changes. No evidence of abnormalities has been shown.

Andrographis paniculata should not be used during pregnancy or lactation. It is contraindicated in cases of known allergy to plants of the Acanthaceae family.

Extracts of Herba Andrographidis may have a synergistic effect with isoniazid.

1. Anti-inflammation of sore throat, relieve of cold. Pills, capsules or tablets are made and taken 3.0-6.0 g four times a day after meals and at bedtime.
    
2. Antidiarrhaeal - The leaves or whole plant are dried and powdered. Pills, capsules or tablets are made and taken 0.5-2.0 g four times a day after meals and at bedtime.
    
3. Treatment of abscesses - The fresh leaves are pounded with water and use as a poultice for abscesses.

As Thai traditional medicine