Bioactive molecules in Siddha Polyherbal Nilavembu Kudineer alleviating symptoms of Dengue/Chikugunya

Rubeena Mattummal, Divya Kallingilkalathil Gopi, Sathiya Rajeshwaran Parameswaran, Sunil Kumar Koppala Narayana*

1 Siddha Central Research Institute, Central Council for Research in Siddha, Ministry of AYUSH, Govt. of India,Arumbakkam, Chennai, India.

Background

Many of the human diseases are cured by modern medicines which sometimes produces unfavorable reactions and toxic side effects. Plants, being a reservoir of medicinal compounds, help in preventing and curing ailments without serious adverse effects [1]. Compounds derived from living organisms, with their significant pharmacological activity, can compete with modern medicines [2]. Plants produce phytochemical constituents for defense against pathogen owing to their characteristic bioactivities. Therapeutic property of every plant is confined to the bioactive compounds present in it. Hence,the screening of these compounds is necessary for the standardization and validation of herbal drugs formed from it. Alkaloids, flavonoids, phenols, terpenoids,tannins and quinones etc. are the important classes of secondary metabolites in plants with significant pharmacological activities. Secondary metabolites derived from plants are reported to possess many important pharmacological characteristics such as anti-oxidant, anti-microbial, anti-allergic, hypoglycemic and anti-cancer properties [3].

Siddha is one of the ancient systems medicines,originating in ancient Tamilakam in south India and Sri Lanka with several polyherbal therapies which were formulated basing on Siddha principles. Siddha system of medicine uses the ancient beliefs and healing methodologies listed by the Siddhars (Siddha physicians)using plants, metals, minerals and various animal products. Diagnosis in this system includes assessing the equilibrium and derangement of the three humors of the body - Vaadham, Pittham and Kapam, the imbalance of which is believed to be the cause of various disease [4].

Dengue is the most prevalent arthropod-borne viral diseases in terms of morbidity and mortality in the recent decade which has re-emerged and remains endemic in more than 110 countries. Two fifths of the world populations (estimating around 100 million) Dengue fever infections, 2.1 million cases of Dengue hemorrhagic fever and 200 thousand deaths worldwide are caused by Dengue every year. Despite extremely high rates of Dengue for decades, Southeast Asia region still recorded an increase of 67% from 1985 - 1989 to 2002 - 2006 [5].Dengue appears in two forms, the classic and severe Dengue. Classic Dengue fever shows symptoms ranging from mild to high fever with retro-orbital pain, severe headaches, maculo-papular rashes, muscle and joint pain.The severe form, Dengue hemorrhagic fever and Dengue shock syndrome may present with abdominal bleeding,hemorrhage and circulatory failure, which is fatal without prompt and proper management [6]. There are four serologic types of Dengue virus (DENV), DENV-1, -2, -3 and -4. Chikugunya is also an arthropod-borne viral diseases with the classic symptoms of fever, joint pain,rash, etc, which is similar to that of Dengue. With the rapid expansion of Dengue and Chikugunya disease in most tropical and subtropical areas of the world, it is crucial to develop effective prevention and control measures, including antiviral drugs and vaccines against them [7].

Siddha medicines are prescribed either as single drug remedy or polyherbal/herb-mineral formulations.Nilavembu k udineer (NK) has been prescribed as a curative and preventive medicine against Dengue [8]. NK is a polyherbal formulation prepared by Andrographis paniculata (Brum.f.) Nees (whole plant), Chrysopogon zizanioides (L.) Roberty (root), Santalum album L. (heart wood), Trichosanthes cu cumerina L. (whole plant),Cyperus r otundus L. (rhizome), Zingiber o fficinale Roscoe (rhizome), Piper nigrum L. (fruit) and Mollugo cerviana (L.) Ser. (whole plant) in equal parts. Kudineer(decoction) is the common name given to the Siddha formulation in which the whole plant (s) or particular part of plant (s) is ground into coarse powder, called as Kudineer Cho ornam (coarse powder for preparation of decoction). It is then made into Kudineer by adding water and heated, so that the mixture of Kudineer Choornam and water reduces to 1/4th or 1/8th of its volume as mentioned in the literature. The dosage to be taken is 30 mL before food, three to four times a day. Lifetime of prepared Kudineer is 1 Samam (3 hours) [9]. The ingredients contain various bioactive compounds like andrographolide, β-vetivenene, α-zingiberene, α-copaene,cyperene, 2-monolinolenin, limonene, β-pinene,β-caryophyllene and α-santalol. The ingredients of the Kudineer possess anti-inflammatory, anti-microbial,analgesic, anti-oxidant, anti-viral, cytotoxic,hepatoprotective and anti-diabetic activities [10]. The anti-inflammatory, anti-viral, and analgesic effects of the various phytochemicals present in NK will help in suppressing and curing the clinical symptoms associated with Dengue. Since the studies about this Kudineer are very scanty, this review aims the documentation of the bioactive compounds and their characteristic pharmacological activities of NK ingredients to throw some light in support of action NK on Dengue.

Andrographis paniculata Burm.f. Nees

Andrographis pan iculata Burm.f. Nees, known as Nilavembu in Siddha, belongs to family Acanthaceae is used traditionally as a remedy against common cold,fever and inflammation, etc. The Indian Pharmacopoeia describes it as a major constituent of at least 26 ayurvedic formulations. In traditional Chinese medicine, it has a significant “cold property” which is used to relieve the body heat, and to drive out toxins from the body [11]. The medical use of Andrographis pa niculata against sore throat has been well known in Thailand [12]. In India, it is used to reduce griping, irregular bowel habits, and loss of appetite of children. Due to its “blood purifying”quality, it is suggested to make use in the treatment of leprosy, gonorrhea, scabies, boils and skin eruptions [13].The key bioactive component of Andrographis paniculata,is the major diterpenoidal constituent, and rographolide.The compound has been reported to have different pharmacological activities like anti-inflammatory [14],anti-cancer [15], anti-microbial [16] and hepatoprotective activities [17] (Table 1). Along with andrographolide the neoandrograpide also imparts the characteristic medicinal activities to the plant and acts as immunostimulant agents that are reported to have both antigen specific and non-specific immune responses [18]. Some studies proved the side effects of andrographolide in creating infertility [19] had led to the questioning of complete safety of NK.

Andrographis p aniculata is an annual herbaceous to arborescent plant with woody branched stem bearing simple, opposite, lanceolate leaves, pink-purple colored,two-lipped zygomorphic flowers and cylindrical to flattened capsules [20]. Beside NK, this plant is the ingredient of other Siddha formulation, Nilavembu camulam [21].

Chrysopogon zizanoids (L.) Roberty

Chrysopogon zizanioides (L.) Roberty is a perennial grass of Poaceae family known for its fragrance oils and medicinal properties. Vetiver oil is composed of more than 170 compounds that are mainly sesquiterpenes and their derivatives (Table 2). Because of the complex nature of the essential oil, it has not been studied intensively[39]. The drug is fibrous, wiry, long, cylindrical roots up to 2 mm in diameter, multi-branched, often attached with stout root stock, smooth or longitudinally grooved, color,light brown, odor strong aromatic, taste and slightly bitter[40]. Veti ver/Vilamiccam ver is used in many Siddha formulations, some of which includes Amirtataik kulikai,Incic choor anam, Maka V acanta kucumakram, Pitta Curak kudineer, Maka e lati kulikai, Nay uruvi nei,Parangi chakkai chooranam [21, 41].

Table 1 Phytochemical constituents in Andrographis paniculata Burm.f. Nees

Cyperus rotundus L.

Cyperus r otundus L. belong to family Cyperaceae is a perennial sedge plant with flowering stem and linear leaves which form a sheath around stem. Rhizome of Cyperus r otundus L known as Koraik kizhangu, is used for the treatment of stomach, bowel disorders, and inflammatory diseases. Drug is a rhizome clothed with flexuous hair, outer surface dark brown, and white inside[47]. The drug is used as anti-inflammatory [48] and anti-malarial [49] (Table 3).

In Ayurveda the dried rhizome is used in dyspepsia/loss of appetite, indigestion, polydipsia/excessive thirst,irritable bowel syndrome, dyspnoea, cough, dysuria,vomiting, lacteal disorders, puerperal disorders, diarrhoea,rheumatoid arthritis and worm infestation [50]. In Siddha Cyperus rotundus is used in many preparations and some are Athimathura mathirai, Adathodai chooranam, Civatai chooranam, Cukku tailam, Ka pada Il akam, S anjeevi theenir. Chandraprakasa mathirai, Kapacurak kudineer,Thathupushti kul ikai, P arangi chakkai chooranam,Milagu thailam [21, 41].

Table 2 Phytochemical constituents in Chrysopogon zizanoides (L.) Roberty

Table 3 Phytochemical constituents in Cyperus rotundus L.

Table 4 Phytochemical constituents in Mollugo cerviana (L.) Ser.

Mollugo cerviana (L.) Ser.

Mollugo c erviana (L.) Ser. is commonly known as Parpadakam in Tamil from the family Molluginaceae. It is an erect slender annual herb with slender cylindrical stem bearing leaves in whorls; flowers numerous and having fruits as rounded capsules, used in Siddha,Ayurveda and folk medicine. The whole plant possesses medicinal properties. The drug is diuretic, anthelmintic,digestive and relieves constipation. It is bitter, cooling and constrictor [65] and used as laxative, stomachic,antiseptic, febrifuge and diaphoretic. It is found to be effective against burning sensation, burning eyes, gastric diseases and fever [20]. The whole plant is used in NK formulation. The pharmacological activities like anti-bacterial [66] and anti-inflammatory [67] actions of Mollugo cerviana have been reported in various studies(Table 4). The plant is also used to prepare Thonthasurak kutinir, a siddha formulation used to cure inflammation,fever and cough [41].

Piper nigrum L.

Piper nigrum L. is a perennial climbing vine of family Piperaceae having flowering woody stem with alternating leaves tapering towards the tip. Dried fruits of Piper nigrum are commonly used in gastrointestinal disorders.These little seeds with enormous health benefits have been an admirable natural remedy for treating various illnesses ranging from obesity to cancer. Fruit is an indehiscent one-seeded globose berry, ovoid to oblong,coarsely, deeply reticulately wrinkled grayish-black in colour, odour, aromatic, taste very pungent [74]. The drug is used as appetizer and is active against helminthiasis,colic [75, 76]. Piper nigrum is used in majority of Siddha formulations and some of them are Amukkara chooranam,Aruvatha mathirai, Iraca ganti meluku, Elatic chooranam,Noccit tailaam, Swasakutori chooranam, Milaku tailam,Vilvati tailam, etc [21, 41]. Piperine, isolated from Piper nigrum has anti-inflammatory, anti-nociceptive, and anti-arthritic property [76] (Table 5).

Santalum album L.

Cantanakkattai, the heart wood of santalum album L, is used as the ingredient drug in many formulations.Sandalwood oil has been widely used in folk medicine for treatment of common colds, bronchitis, skin disorders,heart ailments, general weakness and fever, etc. The heartwood occurs as solid heavy pieces of log or as chips varying in length and width and density. On transversely cut surface it is diffuse porous with medullary ray appearing as fine reddish line [101]. Santalum album L. is used to treat utricaria, diarrhoea and dysentery [102].Santalum album is used in the preparation of formulations such as Arakku tailam, Cintil nei, Cukku ta ilam, Il aku Cantanatit t ailam, Na ciroka Nacat ta ilam, Vallarai nei[21], Kirichannangaluku ennei, K umari t hailam and Mahavilvathi ilakam [41] (Table 6).

Trichosanthes cucumerina L.

Trichosanthes cucumeri na L., known as Peipudal in Siddha, is an annual climber widely used as a medicinal plant in different traditional medicine systems due to its various medicinal values. The fruit is usually consumed as a vegetable due to its good nutritional value [106]. The plant is rich in flavonoids, carotenoids and phenolic compounds which contributes to the antidiabetic,hepatoprotective, cytotoxic, anti -inflammatory, larvicidal effects of the plant. The fruit and roots of this plant contains bryonolic acid, bryononic acid,dihydrocucurbitacin B, chondrillasteryl glucoside and cucurbitacin B [107]. All these active constituents show antiviral characteristics and anti-malarial activity (Table 7). Trichosanthes cucumerina is a climber with palmately lobed leaves with unisexual flowers and ovoid-fusiform fruits [108]. Whole plant is used in the preparation of Cintil ne i, T honthakara kudineer and Manjal n oiku kudineer [21, 41].

Zingiber officinale Roscoe

Known as Chukku in Siddha, Zingiber officinale Roscoe is a perennial herb of family Zingiberaceae with thick rhizome which containing several bioactive constituents,possessing health promoting properties. The characteristic odor and flavor of ginger root come from a volatile oil composed of shogaol and gingerols which make up about 1-3% of the weight of fresh ginger [118]. The main pharmacological actions of compounds isolated include immuno-modulatory, anti-tumorigenic, anti-inflammatory,anti-apoptotic, anti-hyperglycemic, anti-lipidemic and anti-emetic activities [119] (Table 8). The major phytochemicals reported from essential oil of the rhizome are 6-shogaol, 6-gingerol and α-zingiberene. Zingiber officinale Roscoe is considered as a safe herbal medicine with only few and insignificant side effects. Dried drug consists of sympodially branched laterally compressed pieces of horizontal growing rhizome measuring 5 to 12 cm in length, 3 to 5 cm in height and 1 to 2 cm in thickness; the surface is marked with circular closely placed leaf scars, and small circular root scars at places,pale buff to brownish in colour with aromatic odour and pungent taste [74]. Zingiber of ficinale Roscoe has anti-inflammatory, hepatoprotective and antioxidant property in paracetamol induced animal model [120]. The drug is used in colic, haemorrhoids, diseases of throat and inflammation [74]. Dried Zingiber officinale Roscoe is a major ingredient in most of the Siddha preparations and some of them include Agathi ennei, Ka pada i lakam,Pooranathi ilakam, Milaku tailam, Cukku tailam, Nellikai ilakam, Kapa curak kudi neer, Talicati c uranam, Vilvati ilakam, etc [21, 41].

Table 5 Phytochemical constituents in Piper nigrum L.

Table 6 Phytochemical constituents in Santalum album L.

Table 7 Phytochemical constituents in Trichosanthes cucumerina L.

Table 8 Phytochemical constituents in Zingiber officinale Roscoe

Discussion

This review is an attempt to analyze the phytochemicals present in the ingredient drugs of NK so as to see what chemicals reported so far from the ingredients has actions beneficial for treating Dengue and Chikungunya.Essentially a Dengue/Chikungunya medicine must possess properties like analgesic, antipyretic,anti-inflammatory, and antiviral actions to alleviate the reported symptoms such as high fever, headaches, pain behind the eyes, joint and muscle pain, fatigue, nausea,vomiting, skin rash, mild bleeding from nose/gums, bruises,damage to lymph/blood vessels, liver enlargement, and circulatory system failure [142, 143].

Every polyherbal combination in Siddha is based on philosophy of Tridosham (three humours) and Panchamahabhutas (five basic elements) in consideration with the disease conditions. NK was formulated basing on ancient Siddha. But, not any specific bioactives necessarily are responsible for the therapeutic function of each drugs in modern perspective. As the drug is producing beneficial effects in cases of fevers,specifically Dengue and Chikungunya fevers, there is a need to see any connection between the symptoms and bioactivity of the molecules present in each ingredient. As all these eight ingredients are mixed and decoction made by boiling such individualistic action may not be so derogatory in deciding the action of NK. The synergistic action of all these bioactive compounds together may be imparting NK its characteristic medicinal property.

However, from this review, it is evident that these ingredients are used traditionally in the treatment of fever,inflammation, arthritis, gastric ulcer, jaundice and general debility condition while NK is also claimed to have antipyretic, anti-inflammatory, antiviral, and immunomodulatory actions [144]. The bioactives ingredients in combination might be providing relief from body pain, inflammation of the joints and enhancing immune mechanism suppressing the virus. The phytochemicals present in the ingredients also have antiviral and anti-microbial characteristics, which make it suitable medicine for the management of viral fevers. The combination of these ingredient drugs can be substantiated in modern terms on account of its immunostimulant action increasing the defense response in the body which will help an infected person to fight against the viral infection [143].

Potency of b ioactives of NK ingr edients to comb at symptoms of Dengue and Chikungunya

Andrographolide from Andrographis paniculata has been proved to possess anti-inflammatory and hepatoprotective activity, while ninandrographolide has shown immunostimulant property, Β-sitosterol-D-glucoside and wogonin also have anti-inflammatory activities (Table 1).Chrysopogon zizanioides has been reported to possess anti-bacterial and anti-fungal activities, while antiviral activity is yet to be established (Table 2). Cyperenone from Cyperus r otundus possess antiulcer activity;α-cyperone has anti-inflammatory and neuroprotective activity; kobusone has antinflammatory and analgesic activities; isokobuson and iso-cyperol also possess anti-inflammatory effects (Table 3). Mollugo cer viana possesses bioactives with good anti-oxidant potentials which is indirectly beneficial in combating many pathological conditions of body (Table 4). Piperoleine B from Piper nigrum has proved anti-inflammatory and hepatoprotective properties while pipercide has hepatoprotective action, while piperin showed immunostimulant and anti-inflammatory actions (Table 5). Santalum al bum has been reported to possess anti-bacterial and anti-fungal activities, while antiviral activity is yet to be established (Table 6). Cucurbitacin E from Trichosanthes c ucumerina possesses immunomodulatory actions (Table 7). 6-Gingerol, 6-shogaol,hexahydrocurcumin and nerolidol from Zingiber officinale have anti-inflammatory property (Table 8).While aforesaid are from isolated compounds. At extract level, some other properties beneficial in Dengue/Chikungunya like anti-fever (antipyretic) action of Andrographis p aniculata and Mollugo cer viana,anti-emetic property of Cyperus r otundus and Zingiber officinalis and antiviral property of Trichosanthes cucumerina have been proved.

Studies in lines with pr evention of Dengue using ingredients of NK

In a study carried out by Tang et al. [7] for the preliminary screening study for anti-Dengue agent,methanolic extract of Andrographis paniculata was able to maintain most of the normal cell morphologies without causing much cytopathic effect to the DENV-1 infected cells. Thus it possesses high potential to be an anti-Dengue agent, particularly towards DENV-1 serotype. Neoandrographolide, one of the principal diterpene lactones, isolated from a medicinal herb Andrographis p aniculata possesses significant anti-inflammatory effects [145]. The name given to the formulation stands to this ingredient which substantiates the naming.

The aqueous extract of Zingiber officinale may play an important role in the regulation of plasma leakage in Dengue virus infection and decrease the chances of severe Dengue complications by inhibiting the activity and expression of MMP-2 and MMP-9 while upregulating the expression of TIMP-1 and TIMP-2. Therefore, it can help in the development of new anti-Dengue agents [146].

Setback for Nilavembukudinner

Andrographolide a major phyto chemical present in Andrographis p aniculata has been proven to affect spermatogenesis in rats by preventing cytokinesis of the dividing spermatogenic cell lines with appearances of sertoli cell damage and a spermatotoxic effect [19]. The study pointed to a male reproductive toxic effect of a therapeutic use of andrographolide and confirmed the possible prospective use of andrographolide as a male contraceptive. The anti-spermatogenic abilities as well as ovulation preventive effect of the Andrographis paniculata reported may be the main reasons which lead to the controversy of safety of NK as drug. Hence the in-depth studies with standard clinical trials are required to clearly understand the effectiveness of this important formulation and to evade out the misconceptions generated so far. Trichosanthes c ucumerina has been shown to induce liver damage in rats [147]. Very little publication on scientific validations of NK in peer reviewed journals, lack of distinct standard operating procedure for its preparation, deliberative depressing misinformation will be a threat to this valuable traditional medicine.

Conclusion

Bio-actives reported from NK ingredients have shown efficacy for all major symptom of Dengue/Chikungunya.Anti-pyretic, anti-inflammatory, analgesic and immunostimulant effects have been attributed to more than one ingredient in NK. Though these bioactive molecules are present in individual drugs, there might be changes occurring in the structure of these molecules during preparation of Kudineer by mixing and heating all these eight plants with water. Hence more studies have to be undertaken for the scientific evaluation of mode of action of combination of these molecules on fevers like Dengue and Chikungunya. Scientific validation and standardization of the drug is also necessary to maintain its high degree of quality in the global herbal market.

Acknowledgement

The authors extend their heartfelt thanks to Director General, Central Council for Research in Siddha, Chennai for the support.

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