IN VITRO SCREENING FOR ACETYLCHOLINESTERASE ENZYME INHIBITION POTENTIAL OF MUTHU PARPAM – THERAPEUTIC LEAD FOR ALZHEIMER’S DISEASE

Objective: Siddha system is an ancient traditional system of medicine treats many chronic ailments and neurological disorders. Muthu parpam is one of the herbo marine Siddha drugs which have the indication for neurocognitive dysfunction. The main objective of this current study was to evaluate the acetylcholine esterase (AChE) inhibition of Muthu parpam. Methods: AChE activity was evaluated using a modified 96-well microplate assay based on Ellman’s method. Physostigmine (5, 10, 20, and 40 μg/ml) was used as the positive control. Results: The result of this study clearly indicates that the test drug Muthu Parpam was effective in inhibiting AChE enzyme at the specified concentration dose dependently. Maximum percentage inhibition of about 71.68% was observed at 500 μg/ml when compared to that of the Physostigmine, a known AChE inhibitor with the maximum inhibition 84.87% at the concentration of 40 μg/ml. Conclusion: Hence, this preliminary screening has proven the efficacy of Muthu parpam through AChE inhibition potential in the management of Alzheimer disease

ANGIOTENSIN CONVERTING ENZYME INHIBITION POTENTIAL OF ANNAPAVALA CHENDHURAM FOR THE TREATMENT OF HYPERTENSION - AN IN–VITRO ASSAY

Hypertension is that the most noteworthy risk factor for cardiovascular diseases and stroke. Dietary and lifestyle changes play the foremost part to decrease the hazard of hypertension and other related wellbeing complications. Angiotensin Converting Enzyme (ACE) inhibitors play a major role in treating hypertension. Annapavala chendhuram is a herbo – mineral Siddha formulation comes under the type of 32 internal medicines of Siddha. Hypolipidemic activity of Annapavala chendhuram has been proven by some research studies. Hence, the purpose of the present study was to evaluate the ACE inhibition activity on Annapavala chendhuram by using an in-vitro assay. The ACE inhibition assay was evaluated by UV Spectrophotometry technique based on the hydrolysis of histidyl-hippuryl-leucine (HHL) by ACE. About 50µL test sample with varying concentration (100- 500 µg/ml) along with standard captopril (100µg/ml) added with 50µL of ACE and some process had continued. The present study indicates that the test drug Annapavala chendhuram was effective in inhibiting the enzyme ACE dose-dependently. Maximum percentage inhibition of about 53.24±8.403% was observed at 500μg/ml when compared to that of the Captopril, a standard ACE enzyme inhibitor agent with the maximum inhibition 86.98 ± 6.375 at the concentration of 100μg/ml. It was concluded that the test drug Annapavala chendhuram possess significant anti-hypertensive property in protein denaturation assay. So, further in-vitro evaluation of ACE inhibitory activity on Siddha herbal preparations and clinical trials will be the need of the hour

Preclinical Safety Evaluation of "Annapavala Chendhuram”

Raw drug of Annapavala chenduram was procured from the reputed shop in Chennai and got authentication from Department of Geology, University of Madras, Chennai. ❖ As per the literature, APC was prepared in Gunapadam Laboratory, National Institute of Siddha, Tambaram sanatorium, Chennai. ❖ Standardisation was done as per PLIM guidelines. ❖ Physico – chemical analysis of APC revealed its quality standard. ❖ Qualitative analysis of acid and basic radicals for APC showed the acid radicals such as sulphate, chloride, phosphate, carbonate and nitrate. The basic radicals present in the drug were Aluminium, zinc, Iron, calcium, magnesium, potassium and sodium. ❖ The SEM analysis of the drug APC revealed that irregular shape and uniformity of the drug which was 200nm in size. ❖ The EDAX associated with SEM showed the presence of elements like carbon, oxygen, iron, magnesium, Sulphur, Chlorine, Calcium. ❖ The XRD patterns of samples indicated the presence of dominating peaks Fe2O3. ❖ The FTIR analysis of APC revealed that the presence of alcohol, amine, alkane, carbonyl and alkyl halides (Fe- O) group which is ranged from 400 to 4000cm-1. ❖ Animal toxicity was done as per WHO guideline. ❖ There was no mortality or drug related toxicity during acute toxicity study. ❖ The long-term toxicity study did not show any toxic effects in the animals. ❖ Hematological, Serological and histopathology reports revealed the no toxic effects during throughout the study period. CONCLUSION: Annapavala Chendhuram has been used for thyroid dysfunction by many practitioners. The qualitative and quantitative analysis of Annapavala Chendhuram was done as per PLIM guidelines. It revealed that the quality standard of drug. From the animal study, it is concluded that Annapavala chenduram along with proper adjuvant is not toxic on acute administration at a maximum oral dose level of 2000 mg/kg in Wistar rats. There was no mortality and drug related toxicity noted. In long term toxicity, hematological, biochemical and histopathological studies showed no significant adverse effects during the 90 days of the study. Hereby, safety of Annapavala chenduram has been proven through this study. Further efficacy studies and clinical trials on Annapavala Chendhuram will be more beneficial for the researchers and world

Physico-chemical characterization of Annapavala Chenduram - A Siddha herbo marine formulation

Annapavala Chendhuram (APC) is the herbo – marine formulation in Siddha. Annabedhi (Green vitriol) is one of the Uparasam (Hydro chemicals) and Kodipavalam (Coral) is one of the Navamanigal (Nine gems). These both are the main ingredients of APC. Present day demand of Siddha formulations has been raised globally due to increased response towards Siddha system of medicine. Therefore, certain things like standardization, quality control, and safety became essential requirements for Siddha formulations. The test drug APC was prepared at Gunapadam (Pharmacology) lab of National Institute of Siddha, Chennai as per traditional method (incineration) mentioned in the classical texts. Physico‑chemical and instrumental analysis including Scanning Electron Microscopy (SEM), X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) was performed as per PLIM (Pharmacopeia Laboratory of Indian Medicine) guidelines. The pH of test drug APC was 8.34. It contains 0.56% acid insoluble ash and Solubility in the water was 6.75%. Quantitative measures of chloride, sulphate, carbonate, iron, calcium, sodium and potassium was 0.63%, 1.21%, 11.84%, 19.56%, 14.85%, 2.2%, 3.5% respectively. Scanning Electron Microscopy analysis revealed that particle size of APC about 200nm. This study suggests that quality specifications of APC can be developed using parameters described in Siddha along with analytical tools available today.Tex

In-Vitro Acetylcholine Esterase Enzyme Inhibition Potential of Siddha Formulation Vilvaver Chooranam: A Neuroprotective Assay

Background and Aim: Alzheimer's disease (AD) is a progressive neurodegenerative condition evidenced by significant cognitive dysfunction. The state of cognitive impairment is made worse by increased levels of the enzyme acetylcholinesterase (AChE), which is crucial in the hydrolysis of the neurotransmitter acetylcholine (ACh). Siddha therapy gaining higher momentum in recent days due to its global acceptance considering its broad spectral safety and therapeutic window. Siddha originated from the southern geographic landscape of Asia now spreading its wings across the bounders in managing dreadful diseases like AD. The main objective of the present study is to evaluate AChE inhibition of the Siddha formulation Vilvaver Chooranam (VVC). Materials & Methods: In-Vitro Acetylcholine esterase enzyme inhibition Potential of Siddha formulation Vilvaver Chooranam by Ellman’s method. Results:Results obtained from the study clearly demonstrate that the formulation VVC has shown promising acetylcholinesterase at stipulated concentration dose-dependently. Maximum percentage inhibition of about 54.53 ± 3.475 % was observed at 500μg/ml with the IC50 value of 411.9 ± 30.6 µg/ml when compared to that of the Physostigmine, a known AChE Inhibitor with a maximum inhibition 93.44 ± 4.434 % at the concentration of 40μg/ml with the IC50 value of 10.38± 5.29 µg/ml. Conclusion: These findings demonstrate the remarkable potential of these extracts as valuable sources of antioxidants with interesting acetylcholinesterase inhibitory activity.