Solu-Resveratrol (Solu-RSV)

Sub-mircornized Trans-Resveratrol based on SolumerTM technology

Solu-RSV: SolumerTM- Resveratrol Resveratrol (RSV), a polyphenol, naturally present in several

plants such as grapes, was shown to prolong longevity. It’s a small molecule activating the SIRT1 protein, responsible for cell protection against age-related disorders.

The Need: Why the SolumerTM Resveratrol is needed?

Resveratrol is practically insoluble in aqueous media. It demonstrates very low bioavailability and rapid, extensive metabolism, resulting in only trace amounts of unchanged resveratrol in the systemic circulation (Walle, et. al; High absorption but very low

bioavailability of oral resveratrol in humans; Drug metabolism and disposition; 2004; Vol. 32, No. 12; 1377–1382).

Solubest’s Solution:

To answer this need Solubest has developed highly bio-available formulation of resveratrol utilizing Solumer™ proprietary, patent protected technology.

Recent Scientific Evidences: RSV can Influence the Longevity Path through Multiple Mechanisms:

• RSV reduced both gene expression and release of pro-inflammatory cytokines. (The Journals of Gerontology, 2012)

• The beneficial health effects of RSV are mainly mediated by the expression and regulation of deacetylase Sirt1, a protein associated with aging. Recent research in MIT/ Harvard reinforces the claim that resveratrol interacts with key genes in mitochondria. MIT researchers have shown that the mice with normal SIRT1 function given RSV showed dramatic increases in energy (Cell Metabolism, May- 2012) http://hms.harvard.edu/content/more-evidence-longevity-pathway

• Recently, (Feb. 2, 2012) NIH researchers have presented evidence that RSV could exert its function via Sirt1-independent mechanisms: RSV l inhibits certain types of phosphodiesterases, enzymes that help regulate cell energy. Up-regulation of Adiponectin by RSV is also regulated by both the Akt/FOXO1 and the AMP-activated protein kinase signaling pathways and DsbA-L. RSV has a stimulatory effect on the expression levels or the secretion of adiponectin, an adipocyte-derived hormone that plays an important role in the regulation of insulin sensitivity and energy homeostasis, including decomposition of blood sugar and lipid. http://www.jbc.org/content/287/14/11566.abstract?sid=1364a455-e90b-4491-9636-7b953d5fd105

• Significant RSV-induced cardioprotection against ischemia/reperfusion (IR ) injury was determined by recovery to basal level of specific group of microRNA expressions in heart that have altered during IR injuries http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3009730/

• Calorie Restriction-like Effects of 30 Days of Resveratrol Supplementation on Energy Metabolism and Metabolic Profile in Obese Humans. Cell Metabolism, Volume 14, Issue 5, 612-622, 2 November 2011 http://www.cell.com/cell-metabolism/abstract/S1550-4131(11)00386-X

Anti-aging: proposed protective models of action of RSV in aging-associated processes

RSV effects on intracellular signaling associated with prevention of EC activation and unstable plaque development. (Source: Vasc Cell. 2012; 4: 9)

The model predicts that resveratrol, via up-regulating Nrf2-driven antioxidant enzymes and eNOS, down-regulating TNFα-activated NADPH oxidases, exerting mitochondrial protective effects and inhibiting NF-κB, significantly attenuates vascular oxidative stress and inflammation in aging . (Source: Ann N Y Acad Sci.; 2012).

Model of RSV mitichondrial protection:, by activating SIRT1 and/or AMPK, up-regulates Nrf2-driven antioxidant enzymes and eNOS, and down-regulates TNFα-activated NADPH oxidases; by reducing oxidative stress RSV increases the bioavailability of nitric oxide (NO), promoting activation of PGC1α, which increase mitochondrial biogenesis (Source: Exerc Sport Sci Rev. 2011 July; 39(3): 128–132.).

Solumer™: Essential technology characteristics for optimal bio-potency formulation of Solu-RSV

Improved Solubility

Improved Bioavailability

Improved Efficacy

Bottom-up particle assembly

during spray-drying process

Instant powder: upon dispersion in

aqueous media forms nano-colloids

Depressed melting temperature and enthalpy of API

Improved dissolution and solubility in bio-relevant media

Less ordered-crystalline lattice

Submicron particles of API are

homogeneously dispersed in

polymer matrix

SolumerTM is composed of porous micro-

aggregates

Distinct thermodynamic pattern:

favorable for dissolution

Raw resveratrol melting

Formulated Resveratrol melting

• Formulated RSV demonstrates significant reduction in melting temperature and enthalpy in comparison with raw crystalline material.

• Such high energy state requires less energy for dissolution!

Upon contact with water or biological fluids Solumer™ powder is self-dispersed to form uniform nano-colloids with mean particle size in a range of 0.5-5 µm. The colloidal particle is not entire monolith; it is built from subunits of 50-100 nm.

SEM images demonstrates that Solu-RSV is constructed from the porous polymer aggregates (100-200 mm) covered by submicron-micron particles of resveratrol (0.5-5 mm);

Nano- and sub-micron particles of active compound are embedded in polymer matrix

Altered, less ordered-crystalline lattice of stabilized Solu-RSV vs. raw RSV- a distinct feature

0 9 18 27 36 45

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Raw Resveratrol

SoluRes 06-12

Inte

nsity

[co

un

ts]

2 Theta [deg]

Formulated API possesses a high energy crystalline structure, stabilized by interaction with polymers. Such disordered structure reduces the dissolution energy barrier, inherent in the typical rigid crystal of poorly soluble actives!

Instant Powder -upon dispersion in aqueous media forms nano-colloids

Solu-RSV Instant Powder characteristics: • Flowability – easy handling and

processing

• Compressability – allows multiple dosage forms

• Stability – long shelf life

• Bulk instant powder is easy to disperse in liquid

Solubility of native resveratrol and Solu-Resveratrol at 0.1% and 1% concentrations- third party external analysis report:

Solu-RSV - Improved saturation solubility

Saturation Solubility of SoluResveratrol

vs Raw Resveratrol in FaSSIF and FeSSIF

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Raw Resveratrol in FaSSIF

Raw Resveratrol in FeSSIF

SoluRes 02-09 in FeSSIF

SoluRes 02-09 in FaSSIF

5-folds better dissolution in FaSSIF and FeSSIF than raw RSV; similar dissolution profiles in the fed and fasted simulated fluids

The decrease of food dependency is very important feature for the customer compliance.

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Time (min)

Dissolution of SoluResveratrol in FaSSIF vs Resveratrol raw material & Commercially available product

Resveratrol Raw material, 100 mg Res/500 ml FaSSIF

Longevinex, ~100 mg Res/500 ml FaSSIF

SoluRes-02-09 c, 100 mg Res/500 ml FaSSIF

Solu-RSV vs micronized RSV: >2 folds higher dissolution in media simulating intestinal fluid at

fasted state In vitro head to head comparison data shows more than twice higher bioavailability compared to a commercially available micronized Resveratrol and Raw micronized RSV extract in food independent manner

Commercially available ~100 mg/400 ml FaSSIF

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sma

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Time, hr

Solu-RSV (test)

Sito (reference)

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Pla

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Time, hr

Solu-RSV (test)

Sito (reference)

group Cmax

ng/mL AUC0-t

ng.hr/mL

Solu-RSV 407 ± 247 448 ± 334

Sito 71.9 ± 72.2 108 ± 101

P-value* 0.0005 0.0068

group Cmax

µg/mL AUC0-t

µg.hr/mL

Solu-RSV 9.0 ± 2.6 29.3 ± 14.2

Sito 4.6 ± 2.5 25.5 ± 14.3

P-value* 0.0005 0.18

Solu-RSV Clinical Study: ~five folds higher bioavailability

RSV metabolites RSV

Pharmacokinetic Data Analysis: SoluRSV demonstrate >5 times higher Cmax (see below); and 4-10 higher than of the commercially available product references (see data in poster)

Solu-RSV- Product Summary

• Chemical structure:

• Description: Sub-mircornized Trans-Resveratrol based on SolumerTM technology

• Benefit- clinically proven: >four times more bioavailable than conventional extract.

• Structure: Mean particle size of 1-5 microns is composed of clusters of submicron particles

• Patented process and composition: US 6,878,693 (2005), US 7,081,450 B2 (2006), Pending US 2009/0098200

• Dosing: 200-400 mg/ day

Proven Efficacy of RSV- substantial market opportunity

1. Resveratrol Shows Promise to Protect Hearing, Cognition Feb. 20, 2013 http://www.sciencedaily.com/releases/2013/02/130220131742.htm

2. Sport Supplements, wellness: proven fitness accelerator in conjunction with exercise www.sciencedaily.com/releases/2012/06/120619225941.htm

3. Cardiovascular health: cholesterol reduction, vascular support – decrease c-reactive proteins

4. Anti inflammation: inhibition of cyclooxygenase-2-enzyme

5. Anti-aging: protection against free radicals, and longevity – increase of gene expression of SIR2 Longevity Research Raises Hopes, and Questions - NYTimes.com

6. Anti-obesity- Resveratrol pills may mimic effects of exercise and low-calorie diet ...www.guardian.co.uk › ‎Nov 1, 2011

7. Metabolic syndrome, anti-diabetes – inhibition of fat deposition

8. Immune regulation

9. Anti-cancer – antineoplastic activity