Fermentation, Isolation, Structure, and Antidiabetic Activity of NFAT-133 produced by Streptomyces strain PM0324667

Asha A Kulkarni-Almeida,Anagha A Damre,Amrutha S Nair,Pradipta Tokdar,Ram A Vishwakarma,Rajashri R Parab,Saji D George,Manoja K Brahma,Prabhu D Mishra,Girish B Mahajan,Umakant A Bahirat,Kumar V.S Nemmani,Prafull V Ranadive,Prabhu Padmanabhan,Chandni S Thakkar,Nitin J Deshmukh,Amol V Dixit,Nitin V Gaikwad,Lalit S Doshi

doi:10.1186/2191-0855-1-42

Asha A Kulkarni-Almeida, Anagha A Damre + Show 17 more

Open Access

https://doi.org/10.1186/2191-0855-1-42

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Abstract

Type-2 diabetes is mediated by defects in either insulin secretion or insulin action. In an effort to identify extracts that may stimulate glucose uptake, similar to insulin, a high throughput-screening assay for measuring glucose uptake in skeletal muscle cells was established. During the screening studies to discover novel antidiabetic compounds from microbial resources a Streptomyces strain PM0324667 (MTCC 5543, the Strain accession number at Institute of Microbial Technology, Chandigarh, India), an isolate from arid soil was identified which expressed a secondary metabolite that induced glucose uptake in L6 skeletal muscle cells. By employing bioactivity guided fractionation techniques, a tri-substituted simple aromatic compound with anti-diabetic potential was isolated. It was characterized based on MS and 2D NMR spectral data and identified as NFAT-133 which is a known immunosuppressive agent that inhibits NFAT-dependent transcription in vitro. Our investigations revealed the antidiabetic potential of NFAT-133. The compound induced glucose uptake in differentiated L6 myotubes with an EC50 of 6.3 ± 1.8 μM without activating the peroxisome proliferator-activated receptor-γ. Further, NFAT-133 was also efficacious in vivo in diabetic animals and reduced systemic glucose levels. Thus it is a potential lead compound which can be considered for development as a therapeutic for the treatment of type-2 diabetes. We have reported herewith the isolation of the producer microbe, fermentation, purification, in vitro, and in vivo antidiabetic activity of the compound.

Highlights

In 2010, the global prevalence of diabetes was estimated to have reached 285 million and it is predicted to reach 438 million in 2030
We explored the microbial metabolite extract library in search of compounds that enhance glucose uptake in skeletal muscle cells in presence of insulin
We report the antidiabetic activity of NFAT133 along with isolation of its producer microorganism, identification, fermentation, and structure elucidation

Summary

Introduction

In 2010, the global prevalence of diabetes was estimated to have reached 285 million and it is predicted to reach 438 million in 2030. Available agents provide imperfect control of the disease, and the medical need for better therapies is widely recognized (Norman 2010). About 90% to 95% of patients have non-insulin dependent diabetes mellitus (NIDDM) or type-2 diabetes and the standard therapy for the treatment of NIDDM has its own limitations (Mark 1997). Oral hypoglycaemic drugs and insulin are the standard modes of treatment. They are unable to achieve relief from diabetes. This leads to acute and chronic complications. Major efforts have been directed towards development of oral hypoglycaemic drugs, to identify both novel insulin secretagogues and compounds able to enhance insulin action in target tissues

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