Diabetes & its Complications

Author'(s): Raghavan P.R^*

Nanorx Inc, PO Box 131, Chappaqua, NY 10514, US.

^*Correspondence:

PR Raghavan, Founder and CEO, Nanorx Inc, Chappaqua, NY, USA, Tel: (914) 671-0224; E-mail: raghavan@nanorxinc.com.

Received: 30 May 2017 Accepted: 25 June 2017

Citation: Raghavan P.R. Metadichol® A Novel Nano Lipid; GPR 120 Agonist. Diabetes Complications. 2017; 1-3.

Introduction

Diabetes is rampant across the world and was the cause of 4.6 million deaths in 2011. By the year 2030, over 450 million people will be affected [1].

Lifestyle disease modification like diet control can help in preventing Type two diabetes. Many experimental drugs are in various phases of development, but despite new insights into the mechanism of the disease, as of today, no effective solution is in sight.

Increasingly research has been that focussed on the superfamily of GPCRs and there are over 800, seven-transmembrane receptors that display different physiological and pathological functions [2]. Free fatty acids (FFAs) are known to act in regulating physiological functions through G-protein-coupled receptors the most important being GPR-120, also known as FFAR4, which is now an important therapeutic target against diabetes [3].

GPR120 have a ubiquitous presence in various tissues and cells [4]. GPR120 has functions in the homeostatic regulation of systemic metabolism and inflammation depending on this different tissue distribution. GPR120 activation can lead to positive effects on type 2 diabetes. GPR120 signals via both β- arrestin 2 and G proteins in the reported studies. GPR120 is an emerging target in the field of metabolic diseases.

Experimental

The GPCR assay was carried out by DiscoverRX, CA USA using PathHunter® assays which use adapted β-galactosidase complementation system. Enzyme Fragment Complementation (EFC) with β-galactosidase (β-Gal) as the functional reporter. In the PathHunter® β-Arrestin assay. The GPCR assays are the whole cell, functional tests that directly measure activity by detecting the interaction of β-Arrestin with the activated GPCR. It monitors the activation of a GPCR in a homogenous, non-imaging assay format. The enzyme is split into two inactive complementary portions: a small peptide called ProLink™ (PK) and a larger protein, called Enzyme Acceptor (EA). PK and EA are then expressed as fusion proteins in the cell, with PK fused to the GPCR of in-terest, and EA fused to β-Arrestin. When the target GPCR is activated, and β-Arrestin recruited to the receptor, PK, and EA complementation occurs, restoring β-Galactosidase activity which is measured using chemilumi-nescent PathHunter® Detection Reagents. GW9508, a known GPR 120 agonist, was used as the standard.

Cell Handling

• PathHunter® cell lines were thawed from freezer stocks according to standard procedures.
• Cells were seeded in a total volume of 20 μL into white- walled, 384-well microplates and incubated at 37°C for a stipulated time before testing.

Agonist Format

• For agonist determination, cells were incubated with the sample to induce the response.
• Intermediate dilution of samples was carried out to obtain the 5X samples required in assay buffer.
• five μL of 5X sample was added to cells and incubated at 37°C or room temperature for 90 or 180 Fi-nal test vehicle concentration was 1%.

Signal Detection

• Assay signal generated through a single addition of 5 or 15 μL (50% v/v) of PathHunter agent for detection followed by one-hour incubation at room temperature.
• Microplates were then read following signal generation with a Perkin Elmer EnvisionTM instrument for chemi-luminescent signal detection.

Data Analysis

• Compound activity was analyzed using CBIS data analysis suite (ChemInnovation, CA).
• For agonist mode assays, percentage activity was calculated using as follows;

% Activity =100% x (mean RLU of test sample — mean RLU of vehicle control) / (mean MAX control ligand — mean RLU of vehicle control).

Agonist assays, data was normalized to the maximal and minimal response observed in the presence of control ligand and vehicle.

Summary of Results

Table 1 shows the summary of results. Table 2 and 2-1 show the raw data, and Figure 1 shows the graphs of the agonist activity. Metadichol has the same response as the known GPR120 agonist GW9508.

Discussion

Metadichol ® is a Nanoemulsion in water and has a particle size of less than 60 nm. Its mechanism of action is through its binding to the vitamin D receptor (VDR) as an inverse agonist [5].

The natural ligand for the VDR is 1,25-Dihydroxy Vitamin D is and acts as an agonist. Metadichol can act as both positive or as negative agonists on the same receptor a property characteristic of protean agonist [6,7]. In the absence of constitutive activity, it behaves as an active agonist. If constitutive activity is present, it acts as an inverse agonist. Metadichol can reduce or increase insulin secretion [5,8].

For the functioning of the skeletal system [9], Vitamin D is very critical for the regulation of the immune system [9]. It has a vital role not only in controlling insulin synthesis and secretion as well [10,11]. Metadichol in addi-tion to binding with VDR also binds to other nuclear receptors that include PPAR Gamma [12].

Flawed metabolism of Branched Chain Amino Acid (BCAA) is associated with many chronic conditions like type II diabetes and other childhood disorders [13]. Metformin has been shown to in- hibit expression of mitochondrial branched-chain aminotransfer- ase (BCAT) [14]. We have recently shown that Metadichol is a potent inhibitor of BCAT1 [15].

The Agonism exhibited by Metadichol in GPR 120 assays show that it is acting on more than one target, VDR, BCAT1, has been shown previously. The current research approach that of a lock (receptor) and key (drug). The search to mitigate the side effects of drugs by looking for high selective ligands and has proven to be cost and time consuming without any in many ways a failure. Useful drugs act via pathways in targeting multiple proteins rather than single targets. Protein kinase inhibitors like Sutent and Gleevec, have demonstrated that their anticancer actions are due to their effects on multiple kinases [16].

Yildirim, M.A, et.al suggest that there are many keys open a lock but the goal of drug discovery is to have a single key to open many locks, i.e., act via multiple pathways [17]. A practical approach for mitigating disease states may require multiple activities to be efficacious, together with the observation that perturbs biological networks is more important than individual targets [18].

Previously we have published case studies of Metadichol ® in Type 1 diabetes and type 2 diabetes patients [8,19]. It appears to be more efficient than many drugs on the market as it works multiple pathways on multiple receptor targets. Metadichol acts at the nuclear receptor level (VDR) and the transmembrane level (GPR120) and the Cytoplasm and mitochondrial level (inhibition of BCAT1) is probably what makes it such a powerful treatment of choice in diabetes relate diseases.

Conclusion

Metadichol has the potential to serve in mitigating diabetes with a broad spectrum of activity and with a safety that with no toxicity at doses of up to 5000 mg/kg [20-22]. Metadichol is a a far effective substitute to prescrip-tion drugs, which have been largely ineffective in diabetes and have many side effects that add to health care costs.

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