Metadichol ® A Novel Nano Lipid; GPR 120 Agonist

Abstract

Metadichol® is a Nanoemulsion of long-chain alcohols found in many foods. Metadichol acts as an inverse agonist
on Nuclear Vitamin D receptors (VDR) that have a ubiquitous presence in cells and acts by modulating the
immune system and affects many biological processes to modulate many diseases.
We have demonstrated that Metadichol is useful in both type 1 and two diabetes and in modulating insulin levels
and reducing sugar levels and thus increasing insulin sensitivity. We had earlier shown that it binds to VDR and
thus an effect on glucose homeostasis that is a hallmark of VDR pathways. We now report also that it is an
agonist of GPR120 (G protein-coupled receptor 120) which has emerged as a key target for metabolic diseases
like obesity and insulin resistance.
In the in-vitro assay, Metadichol is comparable to GW9508 the most extensively used standard compound in GPR
120 research.

Full text

Volume 1 | Issue 1 | 1 of 4
Int J Diabetes Complications, 2017
Metadichol ® A Novel Nano Lipid; GPR 120 Agonist
Research Article
Nanorx Inc, PO Box 131, Chappaqua, NY 10514, US
*Correspondance:
PR Raghavan, Founder and CEO, Nanorx Inc, Chappaqua, NY,
USA, Tel: (914) 671-0224; E-mail: raghavan@nanorxinc.com.
Received: 15 Apr 2017; Accepted: 02 May 2017
Raghavan P.R
International Journal of Diabetes & its Complications
ABSTRACT
Metadichol® is a Nanoemulsion of long-chain alcohols found in many foods. Metadichol acts as an inverse ag-
onist on Nuclear Vitamin D receptors (VDR) that have a ubiquitous presence in cells and acts by modulating the
immune system and affects many biological processes to modulate many diseases.
We have demonstrated that Metadichol is useful in both type 1 and two diabetes and in modulating insulin levels
and reducing sugar levels and thus increasing insulin sensitivity. We had earlier shown that it binds to VDR and
thus an effect on glucose homeostasis that is a hallmark of VDR pathways. We now report also that it is an
agonist of GPR120 (G protein-coupled receptor 120) which has emerged as a key target for metabolic diseases
like obesity and insulin resistance.
In the in-vitro assay, Metadichol is comparable to GW9508 the most extensively used standard compound in GPR
120 research.
Research Article
Citation: Raghavan P.R. Metadichol® A Novel Nano Lipid; GPR 120 Agonist. Int J Diabetes Complications. 2017; 1-3.
Keywords
Metadichol, VDR, Nuclear receptors, Inverse agonists, Protean
agonists, GPR 120, FFAR4, Lipids, Insulin Resistance, Insulin
sensitivity, Type 1 and Type 2 diabetes, Network biology, Multiple
receptors targeting, BCAT1, PPARG.
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 modication 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 inammation 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 eld 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

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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
• ve μL of 5X sample was added to cells and incubated at 37°C
or room temperature for 90 or 180 minutes. Fi-nal test vehicle
concentration was 1%.
Signal Detection
• Assay signal generated through a single addition of 12.5 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.
Table 1
Compound
Name
Assay
Name
Assay
Format
Assay
Target
EC50 (ug/
mL)
Slope
Curve
Bottom
Curve Top
Max
Response
Metadichol Arrestin Agonist GPR120 3.3141 1.48 -4.1 105 105.3
GW9508 Arrestin Agonist GPR120 3.478 1.03 -5.5 105 101.68
Table 2
Metadichol
Well ID Concentration Raw Value Percent Efcacy
E21 0.048828 16800 -6.3995
E22 0.048828 16000 -8.468
E19 0.097656 17000 -5.8824
E20 0.097656 17600 -4.331
E17 0.19531 16800 -6.3995
E18 0.19531 17000 -5.8824
E15 0.39063 21000 4.4602
E16 0.39063 19000 -0.71105
E13 0.78125 23000 9.6315
E14 0.78125 22800 9.1144
E11 1.5625 29800 27.214
E12 1.5625 27600 21.526
E9 3.125 38200 48.933
E10 3.125 36800 45.314
E7 6.25 48000 74.273
E8 6.25 45000 66.516
E5 12.5 56200 95.475
E6 12.5 55400 93.407
E3 25 62000 110.47
E4 25 58000 100.13
GW6506
Well ID Concentration Raw Value Percent Efcacy
A21 0.0050805 16200 -7.9509
A22 0.0050805 18400 -2.2624
A19 0.015242 16000 -8.468
A20 0.015242 16600 -6.9166
A17 0.045725 17000 -5.8824
A18 0.045725 18800 -1.2282
A15 0.13717 18800 -1.2282
A16 0.13717 18200 -2.7796
A13 0.41152 22600 8.5973
A14 0.41152 21200 4.9774
A11 1.2346 27400 21.008
A12 1.2346 27800 22.043
A9 3.7037 40800 55.656
A10 3.7037 39000 51.002
A7 11.111 49400 77.893
A8 11.111 48000 74.273
A5 33.333 55600 93.924
A6 33.333 59000 102.71
A3 100 61000 107.89
A4 100 56200 95.475
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

Volume 1 | Issue 1 | 3 of 4Int J Diabetes Complications, 2017
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].
Yıldırım, 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
efcacious, 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 efcient 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 with no
toxicity at doses of up to 5000 mg/kg [20-22]. Metadichol is a
a far effective substitute to prescription drugs, which have been
largely ineffective in diabetes and have many side effects that add
to health care costs.
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