Food design in clinical trials: the impact of food formulation on - - PowerPoint PPT Presentation

Food design in clinical trials: the

impact of food formulation

• n absorption and

metabolism of bioactive compounds

Jennifer Ahn-Jarvis Jennifer.Ahn-Jarvis@quadram.ac.uk 2 July 2019

Clinical Research Facility Test Kitchen Quadram Experimental Kitchen OSU Food Science Department OSU Medical Center OSU Agriculture Campus- Wooster

Importance of Oral Disease

• Global Burden of Disease 2015 study

reports that an estimated 3.5 billion people are affected with oral disease1.

• Untreated caries is the most prevalent oral

condition in the world.

• Oral disease is linked to many diseases2,3

1.Kassebaum, N.J. et al J Dent Res 2017;96:380-387 2.Williams, R.C. et al Curr Med Res Opin 2008;24:1635-43 3.Kaye, E.A. J Am Dent Assoc 2007;138:616-9.

https://doctorbrianwest.com/services/m

• uth-body-connection/

Oral Health and Diet

• Diets rich in fruits and vegetables have shown to

promote oral health by improving overall nutrition and microbiome profile1,2.

• Among the many fruits, black raspberries (Rubus
• ccidentalis) have shown in rodent and cell studies

to improve oral health and prevent oral cancer3,4.

• Clinical trials have shown mixed effects

1.Winn, D.M. Am J Clin Nutr 1995 Feb;61:437S-445S 2.Hezel, M.P. and Weitzberg, E. Oral Dis. 2015;21:7-16 3.Casto, B.C. et al Anticancer Res 2002; 22:4005-15 4.Mallery, S.R. et al Cancer Prev Res 2011; 8:1209-21

Oral Residence Time and Oral Metabolites

Enhanced Absorption vs. Enhanced Degradation1

Esophagus Mouth Conjugated-cyanidin Aglycone-cyanidin Mono or disaccharide Cyanidin degradation products

BRB: Freeze-dried black raspberries

1.Ahn-Jarvis et al Cancer Res 2014;74:19

Gut derived metabolites

Dimethyl ellagic acid Ellagic acid Isourolithin A Urolithin C Urolithin D Urolithin B Proposed pathway of ellagic acid metabolism by gut microbiome

• 1. Tomás‐Barberán et al Mol Nutr Food Res. 2017;61:1-35

Ellagitannins

Standardization of Black Raspberry Powder (BRB)

Large Scale Production Black raspberries were procured from Dale Stokes in Wilmington, Ohio (15 kg) and Berri Health, Oregon (6 kg).

HPLC chromatogram of BRB mixture after blending and peak identities verified using external standards and tandem mass spectroscopy1 min 6 7 8 9 10 11 12

mAU

10 20 30 40 50 60 70 520 nm 260 nm 355 nm Cyanindin-3-glucoside Ellagitannin (Sanguiin H-6) Ellagic Acid Rutin Quercetin hexonic acid Ellagic acid- pentose Cyanindin-3-xylorutinoside

1.Gu et al J Agric Food Chem. 2014;62:3997-4006

Dissolution Apparatus to Approximate Oral Residence Time

USP I dissolution apparatus was used to determine oral residence time of various amorphous forms using PBS and synthetic saliva (100 RPM, pH 6.5 at 37°C)6,7.

0% 20% 40% 60% 80% 100% 50 100 150 200 250 300

Percentage of Total Phenolic Released

Time (minutes)

Hard Candy Saliva Hard Candy PBS Starch- Saliva Pectin- Saliva Pectin PBS Starch PBS

Three amorphous forms show 3 distinct release behaviors1 *Complete dissolution occurred at 600 minutes. Different letters denote differences in rate of total phenolic release using ANOVA (p≤0.05) with Tukey’s posthoc test

a b c*

Hard Candy (glassy) Pectin (visco-elastic) Starch (elastic)

slow fast intermediate

1.Gu et al J Food Sci. 2015; 80:E610-8

Sensory Evaluation for Clinical Adherence and Optimization of Formulation

Sensory Evaluation confections conducted under sensory booth conditions (n=65). Letters denote mean separation where significant differences (p≤0.05) were found using ANOVA with Tukey’s posthoc test. 1 2 3 4 5 6 7 8 9

Overall Liking Color Aroma Fruit Flavor Sweetness Texture

9 Point Hedonic Scale Attributes

Hard Candy Pectin Starch

Dislike Extremely Like Extremely

a ab b a a b a b b a c b a b ab a a a

Optimized Confection Formulations

BRB: Standardized freeze-dried black raspberry powder Starch: Tate and Lyle Confectioners G cornstarch Pectin: Danisco Grindsted CF 130B

Composition (g) Hard Candy Pectin Confection Starch Confection BRB powder 1.25 1.25 1.25 Sugar 3.15 2.18 0.63 Corn syrup 1.85 0.75 2.28 Water

• 1.85

1.70 Starch/pectin

• 0.09 (pectin)

0.39 (starch) 50% (w/w) citric acid

• 0.13
• Total

6.25 6.25 6.25

Study Design

n = 10 n = 10 n = 10 n = 10 n = 10

67 Adults (33 men and 34 women)

18 to 65 years old BMI 18 to 35 kg/m2 Never or past smokers for >10 years

R A N D O M I Z E

Berry Confection 2: Hard Candy Dose 1 (3 pieces / day) Berry Confection 2: Hard Candy Dose 2 (6 pieces / day) Berry Confection 2: Pectin Dose 1 (3 pieces / day) Berry Confection 2: Pectin Dose 2 (6 pieces / day) Berry Confection 3: Starch Dose 1 (3 pieces / day) Berry Confection 3: Starch Dose 2 (6 pieces / day) Washout (2 weeks) Low phenolic diet Intervention (2 weeks) Day -14

• consent and enrollment
• instruction of washout (restricted low

phenolic diet)

• multivitamin regimen

Day 0

• buccal scrape
• urine collection
• blood collection
• Durability Test

Day 14

• buccal scrape
• urine collection
• blood collection

n = 10

Demographic and Adherence Measures of Confection Study

Age (mean ± SD)

33 ± 11 years old

BMI (mean ± SD)

26.6 ± 9.2 kg/m2

Compliance (mean ±SD) Hard Candy

93.2 ± 15.0%

Pectin

93.9 ± 12.8%

Starch

96.4 ± 12.3%

History of dental surgery

13% (9/67)

Twice daily teeth brushing

66% (44/67)

Daily flossing

69% (46/67)

Regular alcohol consumers (average 3.25 servings/ week)

69% (46/67)

Never smokers

89% (60/67)

Past smokers (0.5 pack/ 5.5 years)

11% (7/67)

Oral Residence Time (Assessment of Confection Durability)

• Three confections were administered in a

randomized crossover design during a single visit.

• Participants

were instructed to vigorously tumble confection but prohibited from chewing confection.

Each subject was asked to record start and stop times for each confection

*Oral residence time in minutes (A) and quantity of saliva produced with each confection (B). Letters denote differences using ANOVA (p≤0.05) with Tukey’s posthoc test

Salivary Metabolites

Different letters denote significant (p≤0.05) differences usingANOVAwith Tukey’s posthoc test

1 2 3 4 5 6 7 8 9

Cyanidin-3-glucoside

Total Anthocyanins in saliva collected

b a b Maximum total cyanidin in confection (8.5 ± 0.6 mg)

0.E+00 1.E+05 2.E+05 3.E+05 4.E+05 5.E+05 6.E+05 7.E+05

pca pga

Relative abundance (mAU) hard pectin starch

24 hour Total Urolithins

• 3 confections at two doses (4

and 8g/day) consumed for 14 days (n=62).

• Total urolithin reflects urolithin

A to D and isourolithin A.

Different letters denote mean separation where significant differences (p≤0.05) were found in total urolithin in 24 hour urine using ANOVA with Tukey’s posthoc test

Conclusions

• 1. Oral residence time: hard candy was the most durable having longest oral

residence time resulting in the greatest volume of saliva

• 2. Salivary metabolites: cyanidin-3-glucoside significantly less during hard candy

consumption but ellagitannin and ellagic acid increased.

• 3. Huge inter-individual differences observed in the rate of confection

consumption, quantity of saliva produce, and polyphenolic profile

• 4. 24 hour urine: No differences in total urolithins excreted with 4g dose but

significant differences were observed between pectin and hard candies with 8g dose of BRB suggesting that confection matrix may impact ellagic acid metabolism

Acknowledgements

College of Agriculture and Food Sciences Department of Food Science and Technology Ken Riedl, Ph.D. Steven J. Schwartz, Ph.D. Matthew Teegarden, M.S. Yael Vodovotz, Ph.D. College of Medicine Division of Internal Medicine Steven K. Clinton, M.D., Ph.D. Department of Biomedical Statistics Denis Pearl, Ph.D. College of Pharmacy James Fuchs, Ph.D. College of Public Health – Division of Environmental and Health Sciences Christopher Weghorst, Ph.D. Thomas Knobloch, Ph.D. Steve Oghumu, Ph.D. Center for Advancement of Functional Foods and Entrepreneurship Sensory and Clinical Studies Volunteers CTOC Postdoctoral Fellowship National Institute of Dental and Craniofacial Research (T32 DE014320). Comprehensive Cancer Center and the National Cancer Institute (P30 CA016058). Center for Clinical and Translational Science and the National Center For Advancing Translational Sciences (UL1TR001070). United States Department of Agriculture:USDA 38903‐03560