Grant e du bois

Grant E DuBoisAdvances in the Science of

Sweet Taste & Sweetness Enhancement

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Advances in the Science of


Advances in the Science of


Grant E DuBoisResearch Fellow

The Coca-Cola CompanyAtlanta, GA

USA

The Coca-Cola Company

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OutlineOutline

I. Sensing Foods and Beverages

II. Metrics for Commercial Viability of Sweeteners and Sweetness Enhancers

III. Sweetness Modulation, aka Optimizing the Tastes of the Non-Caloric Sweeteners of Today

IV. Sweetener Receptor Positive Allosteric Modulators, a Novel Approach to Sugar-Like Taste


The Coca-Cola Company 4

I. Sensing Foods & Beverages:

Gustation

I. Sensing Foods & Beverages:

Gustation

The Coca-Cola Company5

Gustation: A Superimposition of 5 Taste Modalities

Gustation: A Superimposition of 5 Taste Modalities

1. Sweet2. Bitter3. Umami 4. Salty5. Sour


The Biology of GustationThe Biology of Gustation


Lingual Taste Buds are Found in PapillaeLingual Taste Buds are Found in Papillae


Are You a Non-Taster, a Taster or a Supertaster?Are You a Non-Taster, a Taster or a Supertaster?

Linda Bartoshuk, U of Florida


What Do Taste Buds and Taste Bud Cells Look Like?What Do Taste Buds and Taste Bud Cells Look Like?

M. Behrens, W. Meyerhof, C. Hellfritsch and T. Hofmann, Angew. Chem. Int. Ed., 2011, 50, 2220-2242.


How do Sweeteners, Bitterants and Savory Compounds Excite Taste Bud Cells?

How do Sweeteners, Bitterants and Savory Compounds Excite Taste Bud Cells?

Endoplasmic Reticulum

Receptor

Electrical Message to Brain

Sweetener/Bitterant/

Savory Compound

Saliva

Phospholipase C

Ga

Gb

GgGDP

GTP

Cytosol~-60mV

IP3 Ca2+

IP2

Gb

Gg

TRPChannel

Na+

Na+

Na+

Na+

Na+

The Coca-Cola Company 11

What does the Sweetener Receptor Look Like and How does Sugar Activate it?

What does the Sweetener Receptor Look Like and How does Sugar Activate it?

T1R2 T1R3

Sucrose

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II. Metrics for Commercial Viability of

Sweeteners and Sweetness Enhancers

II. Metrics for Commercial Viability of

Sweeteners and Sweetness Enhancers


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In order to realize outstanding commercial success, a new sweetness technology must deliver on all of

the metrics........Safety, Taste Quality, Stability, Solubility and Cost …… and without compromise

on Safety or Taste Quality.

In order to realize outstanding commercial success, a new sweetness technology must deliver on all of

the metrics........Safety, Taste Quality, Stability, Solubility and Cost …… and without compromise

on Safety or Taste Quality.


Key Message: Key Message:

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Commercial Viability is Dependent on 6 Metrics

Commercial Viability is Dependent on 6 Metrics

1. Safety2. Taste Quality• C/R Function & Rm

• Flavor Profile• Temporal Profile• Adaptation Profile

3. Stability4. Solubility5. Cost 6. Patentability

G. E. DuBois, “Sweeteners and Sweetness Modulators: Requirements for Commercial Viability”, Chapter 29 in Sweetness and Sweeteners, G E DuBois and D Weerasinghe, Eds., ACS Symposium Series 979, ACS Books, Washington DC, 2008.


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III. Sweetness Modulation, aka

Optimizing the Tastes of the Non-Caloric Sweeteners of Today

III. Sweetness Modulation, aka

Optimizing the Tastes of the Non-Caloric Sweeteners of Today


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1. HP sweeteners known today do not match the Taste Quality of sugar……….and delivery on the Taste Quality metric requires “Sweetness Modulators”; and

2. Since trial-and-error approaches have been employed for saccharin (>100 years) and aspartame (~30 years) w/o significant progress……..a new paradigm……..a Mechanism of Action (MOA) based paradigm………is needed to find effective “Sweetness Modulators”.

1. HP sweeteners known today do not match the Taste Quality of sugar……….and delivery on the Taste Quality metric requires “Sweetness Modulators”; and

2. Since trial-and-error approaches have been employed for saccharin (>100 years) and aspartame (~30 years) w/o significant progress……..a new paradigm……..a Mechanism of Action (MOA) based paradigm………is needed to find effective “Sweetness Modulators”.


Key Messages: Key Messages:

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MOA Approach to Rebaudioside A Sweetness Modulators

CH2CH3

H

H

O

O

OO

H3CHO

OH

OH

HO

O

O

O

HO

OH

O

OH

OH

HO

HO

O

HO

OH

OH

OH

H

H

H

H

Operating Principle: The non-sugar-like taste of REBA is intrinsically linked to its chemical structure!


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MOA Approach to Rebaudioside A Sweetness Modulators (Cont’d)

• Maruzen Pharmaceutical Company: NaCl found to cause a 2.5-fold increase in potency of stevia sweeteners.

• The Coca-Cola Company: NaCl, and/or other osmolytes, found to cause increases in sweetness potencies as well as elimination of the slow sweetness onset and the sweetness linger for Rebaudioside A and other HP sweeteners thus leading to the Non-Specific Binding Hypothesis.1, 2

1. I. Prakash, G. E. DuBois, P. Jella, G. A. King, R. I. San Miguel, K. H. Sepcic, D. K. Weerasinghe, N. R. White, US Patent Application 20070128311 (June 7, 2007).2. G. DuBois, “Validity of early indirect models of taste active sites and advances in new taste technologies enabled by improved models” Flavour and Fragrance Journal 2011, Electronic Version: wileyonlinelibrary.com (DOI 10.1002/ffj.2042), Print Version: July-August Issue.

• Keiko Abe Laboratory (University of Tokyo): Support for the Non-Specific Binding Hypothesis as the rationale for slow sweetness onset and sweetness linger.3

3. K. Abe, et al., “Surface Plasmon Resonance Analysis on Interactions of Food Components with a Taste Epithelial Cell Model”, J. Agric. Fd. Chem. 2010, 58 (22), 11870-11875.


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MOA Approach to Rebaudioside A Sweetness Modulators (Cont’d)

OsmolyteConcentration (mOsM)

Sample A Sample B Sample C Control

Erythritol 500 0 0 0

NaCl 0 500 0 0

KCl 0 0 500 0

Osmolarity (Total) 500 500 500 0

Sweetness Linger (0-5) 0 0 0 5


Rebaudioside A (500mg/L) / Osmolyte Formulations

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IV. Sweetener Receptor Positive Allosteric Modulators(aka Sweetness Enhancers),

a Novel Approach to Sugar-Like Taste

at Lower Calories

IV. Sweetener Receptor Positive Allosteric Modulators(aka Sweetness Enhancers),

a Novel Approach to Sugar-Like Taste

at Lower Calories


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Sweetener receptor positive allosteric modulators (PAMs)…..……aka enhancers…….which

potentiate the activities of carbohydrate sweeteners enable accurate reproduction of carbohydrate

sweetener taste quality w/ dramatic reduction in caloric content.

Sweetener receptor positive allosteric modulators (PAMs)…..……aka enhancers…….which

potentiate the activities of carbohydrate sweeteners enable accurate reproduction of carbohydrate

sweetener taste quality w/ dramatic reduction in caloric content.


Key Message: Key Message:

Sweetener Receptor Positive Allosteric Modulators

An Early Conceptual View of the Function of a Positive Allosteric Modulator (PAM) in the Sweetener Receptor

22The Coca-Cola Company

Receptor Binding

Site

SucrosePAM

Receptor Binding

Site

Receptor Binding

Site

Sucrose

Receptor Binding

Site

PAM

Sucrose

SucrosePAM

Signal: 0

Signal: 1

Signal: >>1Signal: 0

Proof of Concept for Sweetness Enhancement

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O

OH

O

H

HO

2001: @ 375 mg/L, DHB enhances 6% sucrose to 8% sucrose w/o any off taste! A weaker effect was noted for fructose and no effect for glucose.

G. DuBois, “Validity of early indirect models of taste active sites and advances in new taste technologies enabled by improved models” Flavour and Fragrance Journal 2011, Electronic Version: wileyonlinelibrary.com (DOI 10.1002/ffj.2042), Print Version: July-August Issue.

DHB


Senomyx Collaboration: PAM Discovery by HTS / Cell-Based Assay

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~8-Fold Sucralose Enhancer

G. Servant, C. Tachdjian, X. Tang, S. Werner, F. Zhang, X. Li, P. Kamdar, G. Petrovic, T. Ditschun, A. Java, P. Brust, N. Brune, G. E. DuBois, M. Zoller and D. S. Karanewsky, Proc. Natl. Acad. Sci. USA, 2010, 107, 4746-4751.

R. Shigemura, C. Podgurski, B. Kitisin, J. Ward, A. Alatorre, PCT Patent Application No. WO2010/014813 A2 (February 4, 2010).

N

SN

O NH

HO

O

HCH3H3C

O

NHH3C

N

NS

H3C

H3C NH

O

H

H

~2-Fold Sucrose Enhancer



T1R2 T1R3

PAM

Sucrose

A B

MOA for PAM Activity at the Sweetener Receptor

Adapted from S. D. Munger et al.

Sweetener Receptor PAMs

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• Original Hypothesis: CHO Sweetener / PAM formulations will taste equivalent to CHO Sweeteners (i.e., not be limited by low Rm, bitter, metallic, licorice, etc. off tastes, slow sweetness onset / sweetness linger and sweetness adaptation; observations to date support that this hypothesis is correct!


• Wood et al. 2011 Review:* “A PAM will exert its effects only when the endogenous agonist is present, resulting in temporal and spatial activity of the endogenous ligand.”

*M. R. Wood, C. Hopkins, J. T. Brogan, P. J. Conn and C. W. Lindsley, Biochemistry 2011, 50, 2403-2410.

What will be the Future of Sweetener Receptor PAMs?

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• Efficacy of Sweetener Receptor PAMs: Ma et al.,* have reported GPCR PAMs w/ >100-fold enhancements. Can 100-fold enhancers be found for the sweetener receptor?

*L. Ma, M. Seager, M. Wittmann, M. Jacobson, D. Bickel, M. Burno, K. Jones, V. Kuzmick Graufelds, G. Xu, M. Pearson, A. McCampbell, R. Gaspar, P. Shughrue, A. Danziger, C. Regan, R. Flick, D. Pascarella, S. Garson, S. Doran, C. Kreatsoulas, L. Veng, C. W. Lindsley, W. Shipe, S. Kuduk, C. Sur, G. Kinney, G. R. Seabrook, W. J. Ray, Proc. Natl. Acad. Sci. USA 2009, 106, 15950-15955.

• Agonist Specificity for Sweetener Receptor PAMs: PAMs reported to date exhibit specificity for individual CHO sweeteners. Can PAMs be identified which enhance all sweeteners?

• Natural PAMs: PAMs reported to date are synthetic compounds which are being developed as artificial flavors. Can commercially-viable natural enhancers be found which can be labeled as natural flavors?

• PAMs for Natural High-Potency Sweeteners: PAMs for stevia sweeteners would enable their use at lower concentrations which may help address their negative taste attributes. Can commercially-viable enhancers for stevia and other natural high-potency sweeteners be found?


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