Abstract
Aim: Synthetic lead molecules are associated with host of adverse effects while medicinal molecules isolated from natural sources are blessed with both safety as well as efficacy. The ancient doctrine of Ayurveda ardently advocates the therapeutic virtues contained in green leaves of Spinacia oleracea L. The principal constituent of the leaves is the class of metalloporphyrin chlorophyll, which is also the floral counterpart of faunal heme. Chlorophyll-a (Chl-a) and chlorophyll-b (Chl-b) are the cardinal members of the chlorophyll family.
 Study design: Herein, we have explored the anti-radical, cytoprotective, anti-inflammatory and anti-proliferative efficacy of Chl-a and Chl-b in reference to standard drug and crude extract of Spinacia leaves. The current study is aimed to establish, naturally mined metaloporphyrins as safe and efficacious replacement of synthetic leads that are associated with a wide range of toxicological issues.
 Methodology: Using a combination of Silica Gel-G column chromatography and preparative thin layer chromatography, the two principal green metallo-porphyrins (Chl-a and Chl-b) were sequentially extracted and isolated from crude extract of Spinacia oleracea L leaves. Antiradical efficacy, of the isolated green porphyrins was quantified by DPPH and Hydrogen peroxide radical scavenging assay. Cytoprotective efficacy was evaluated using ex-vivo hemolysis assay and anti-inflammatory potency was attested employing carrageenan induced paw edema bioassay. To enumerate on the anti-proliferative potency, MTT assay was employed, while toxicology of the isolates was evaluated employing OECD 420 acute toxicity guidelines.
 Findings: The study confirmed that isolated green porphyrins Chl-a and Chl-b as well as crude extract all exerts significant anti-radical, cytoprotective, anti-inflammatory and anti-proliferative efficacy however while potency of Chl-a was at par with that of reference standard and superior to the crude extract, Chl-b clocked in a value inferior to both. Furthermore, acute toxicity study indicated that even at p.o. dose of 2000mg/Kg b.w, no toxicity was manifested in either of the metalloporpyrin treated groups thus ascertaining the safe nature of the naturally mined metalloporphyrin entities. Also naturally mined Chl-a is not only a safer alternative to synthetic medicine but it is more potent and safe than its parent extract popularly used in herbal medicine.
 Conclusion: The results of the study indicates that Chl-a having a more profound structural resemblance to heme than Chl-b can be further modulated as a cost-effective and safe anti-radical alternative to synthetic leads in inhibiting inflammation and untoward cell proliferative while extending cyto-protection from pathological ROS generated in diseased states.
Highlights
The major limitation of synthetic lead molecules is their association with a wide range of untoward reactions [1]
The study confirmed that isolated green porphyrins Chl-a and Chl-b as well as crude extract all exerts significant anti-radical, cytoprotective, anti-inflammatory and anti-proliferative efficacy while potency of Chl-a was at par with that of reference standard and superior to the crude extract, Chl-b clocked in a value inferior to both
The crude extract obtained from Spinacia leaves contained a rich assortment of compounds with approximately nine different spots on the TLC chromatogram (Fig. 2)
Summary
The major limitation of synthetic lead molecules is their association with a wide range of untoward reactions [1]. In the current study we have attempted to establish the therapeutic utility of naturally occurring metallo-porphyrins in reference to both synthetic reference drug as well as its parent crude extract. Doctrine of signature exploits the structural similarities existing between the floral and the faunal kingdom, so as to establish the pharmacological significance of herbs and plant derivatives in traditional medicine [3]. While heme is quintessential for the physiological sustenance of human beings, chlorophyll is acknowledged as the key pigment of the photosynthetic realm. It predominantly aids in production of oxygen necessary for fulfilling the various myriad needs of the animal kingdom [5]. Complete enzyme set for structurally novel antimutagenic flavonoids chlorophyll biosynthesis in Escherichia coli
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