Preclinical and Clinical Evaluation of Safety and Efficacy of Siddha Herbo Mineral Formulation Nandhi Mezhugu in the treatment of Uthira Vatha Suronitham (Rheumatoid Arthritis)

The study drug Nandhi Mezhugu(NM) was tested for preclinical and clinical study, The findings of the drug were given as follows: • The raw drugs for NM were purified and the medicine NM was prepared at IMPCOPS as per the SOP mentioned in Siddha Vaidhiya Thirattu. • The physico-chemical analysis of the drug revealed that the drug possessed high value for loss on drying , total ash indicated the presence of inorganic substances and the water soluble ash value showed the presence of water soluble inorganic salts like sodium chloride. The water soluble and alcohol soluble extractives indicated the presence of high polar secondary metabolites like glycosides, sugars, tannins, saponins and alkaloids. The calculated acid value, saponification value and iodine value were indicative of purity of the ghee used for the preparation and showed the number of milligrams of free acids and saponifiable unsaturated acid in the drug. The shelf life of the drug is increased. • The phyto-chemical analysis of the extract of Nandhi mezhugu revealed the presence of secondary metabolites, viz., Steroid, Triterpene, Flavonoid, Alkaloids, Phenol, Tannin, Saponin, Coumarin, Glycoside, Quinone, which improve the therapeutic efficacy of the drug. • The qualitative inorganic analysis of the drug showed the presence of mercury, magnesium, aluminium, calcium, sodium, potassium, copper, zinc, iron, cobalt, chloride, carbonate, nitrate, sulphate, sulphide, acetate, silicate and arsenate which are biologically important radicles. • Quantitative results of heavy metal analysis revealed that the contents of lead and cadmium were within the admissible limits, whereas the contents of arsenic and mercury were high due to the presence of the ingredients Rasam (mercury), Lingam (Mercuric sulphide), Pooram (Mercurous chloride), Thalagam (Arsenic trisulphide), Manosilai (Arsenic disulphide), Vellai padanams (Arsenic trioxide), gowri padanam(Arsenic penta sulphide), Kalmatham (Hydrous cobalt arsenate) and Rasa chenduram/sinduram (Red sulphide of mercury). • Quantitative assays showed the presence of Calcium, Magnesium, Potassium, Aluminium, Copper, Iron and Zinc . Sulphur (as SO2) and Chloride (as NaCl). • In the microbial study, the drug was found free from E. coli, Salmonella spp., Staphlococcus aureus and Enterobacteriacea. • All the four aflatoxins were not detected in the drug. • Various pesticide residues were lower than the limit of quantification. CONCLUSION: The raw ingredients of the study drug, Nandhi Mezhugu (NM), were purchased and prepared at the Indian Medical Practitioner‘s Cooperative Pharmacy (IMPCOPS) society, as per standard operating procedure mentioned in the text Siddha Vaithiya Thirattu and standardised as per Ayush guidelines. The safety studies - acute, sub-acute, sub-chronic of NM, done as per OECD guidelines.Acute toxicity findings showed no mortality and toxicity signs up to 2000 mg/kg in acute oral toxicity study. NM can be classified under category-5, since the LD50 value was greater than 2000 mg/kg in accordance with Globally Harmonized System of Classification and Labelling of chemicals (GHS 2015] and this provided us a direct relevance for protecting human and animal health. NM did not produce delayed onset of toxicity in both 28 days and 90 days repeated oral toxicity studies. Based on these results, No Observed Adverse Effect Level (NOAEL) of ―NM‖ was greater than 110 mg/kg/day. ICP-OES –study for detection of heavy metal traces in animal tissue samples (Brain, Liver, Kidney) in Sub-chronic toxicity studies (High dose group) of NM reported the absence of traces of heavy metals such as lead, cadmium, mercury and arsenic. NM showed significant analgesic and anti-inflammatory (acute and chronic) and anti- arthritic activities in animal models. In vascular permeability study, NM caused a significant decrease in vascular permeability. In in-vitro cytotoxicity assay, in a LPS induced inflammation in the RAW 264.7–Mouse macrophage cell line study, NM showed cytotoxicity at 500 μg/ml and above. NM at 100 μg/ml showed a decrease in inflammatory markers to the extent of 14% and 10% of COX-2 and TNF-alpha respectively. This result suggests that the Nandhi Mezhugu preparation can be used as an alternative treatment for the inflammatory conditions. The pro-inflammatory and anti-inflammatory gene expression studies performed in the rat paw treated with Nandhi Mezhugu, showed a decrease in C-reactive protein and Interferon-gamma gene expression which is considered due to anti-inflammatory effect of Nandhi Mezhugu. An open labelled, non-randomized, clinical study (without control) was conducted in 40 cases of Uthiravatha suronitham (or Rheumatoid arthritis), to determine the safety and therapeutic efficacy of Nandhi mezhugu (NM). The study drug NM was given for 60 days (7 days drug dosing followed by 7 days drug holiday). At the baseline, 100% of both male and female patients were tested high positive for Anti CCP, RAF and high ESR, and raised CRP level. After 60 days of treatment, the haematology report showed a significant reduction in the high value of ESR (****p value -0.0001). While comparing the RAF before and after treatment, a significant reduction was observed (*p value-0.032). The quantitative outcome of MHAQ score, Universal pain assessment scale score, Restricted movement assessment scale score and EULAR score showed significant reduction after treatment and proved the therapeutic efficacy of NM for the treatment of RA