Boost Oral Bioavailability of Polyphenol

It is considered that less than 5% of the dietary polyphenols are absorbed as its aglycone form (generally recognized as active form that brings bioactivity). Hughes has developed “RenoSorb” innovative fermentation technology platform, the best solution for greatly enhancing the aqueous solubility and oral bioavailability of polyphenols through bioconversion by GRAS bacterial.

First in the world, Hughes combines pharmaceutical phosphate pro-drug strategy and polyphenols and makes them become totally water soluble after a hydrophilic phosphate group addition. Our pro-polyphenols can easily transfer the natural gut barrier 'unstirred water layer', then quickly hydrolyzed by an ubiqitous enzyme 'alkaline phosphatase' located on the apical epithelial intestine cells, and give the fast and real absorption to systemic circulation maintaining as its original aglycone form.

One of our ingredients, RenoCidin, the fermented citrus bioflavonoid containing highly water-soluble hesperetin phosphate, showing 100,000X higher water solubility and 6X higher bioavailability than commercial hesperidin, has already been approved by USFDA as new dietary ingredient (NDIN #1253).
Another ingredient, RenoFlavone, the fermented soy isoflavones containing highly water-soluble genistein phosphate, also showing 100,000X higher water solubility and 3.7X higher bioavailablity than commercial genistein. The IP has been granted in six countries including EU, US, JP, KR and TW, which related to prevention and treatment of postmenopausal osteoporosis.

Through RenoSorb technology application, Hughes's polyphenol ingredients totally break the oral absorption limits and maximizes their efficacy, eat less but get more. It endues polyphenols with efficacy, efficient and to an exciting new level.

1. Hughes Biotech. Co. Ltd.
Hughes Biotech
Our Innovation is Your Solution
Next Generation Functional Ingredients
–soluble form for maximum efficacy

2. Company Profile
2
Company Name :Hughes Biotechnology Co., Ltd.
Head Office :12F.-2, No.420, Sec.1, Keelung Rd., Xinyi
Dist., Taipei City 11051, Taiwan
New Groups Founded :2017
Number of Employee :10
Business :1.RenoSorb
TM
TechnologyPlatform
across nutraceutical,
pharmaceutical, and skincares.
2.Development, manufacture and
sales of novel polyphenolic
ingredients
Subsidiary Company :Hughes Animal & Nutrition Limited Co.
HUGHES

3. Our Location
3
3
Taiwan is here
Taipei City-Taipei101 National Taiwan University

4. History of Hughes Biotechnology
4
•Licensed the patented
technology from National
Taiwan University.
•Established the optimal
lab-scaled process of
RenoSorb
TM
technique.
2017- 2019First three product pipeline
•RenoFlavone
TM
•RenoCidin
TM
•RenoSage
TM
2020
2021
•RenoCidin
TM
US FDANDINNo.1253
(New Dietary Ingredient Notification)
2022
2023
•RenoFlavone
TM
•RenoSage
TM
US FDANDINo.1331 (pending)
•RenoVital
TM
coming soon…
2024
•RenoFlavone
TM
US FDA NDIN No.1283
•FINALISTof Vitafoods Europe Startup Innovation
Awards “The most innovative technology or
service supporting nutraceutical industry.”
•Established distribution contracts spanning
across the US, KR, and TW.
United States, European Union,
Japan, Korea, Taiwan patents
“Method of treatment or
prevention of osteoporosis”

5. Structural diversity of polyphenols and distribution in foods
5
gallic acid
O
O
O
OH
OH
OCH
3
OH
O
OOH
OH
OH
O
O
OH
OH
OH
O
OH
O
OH
OH
OH
OH
O OH
OH
OCH
3
OH
OCH
3
OOH
OH
OH
OH
O
+
OH
OH
OH
OH
OH
O
O
O
O
O
O
H
H
OH
OH OH
O OH
H
3
CO
OH
OH
O
Polyphenols
Flavonoids
Non-flavonoids
Lignans
Curcumin
Hydroxycinnamic acidHydroxybenzoic acid
Phenolic acids
Flavonol Flavone Flavanol Flavanone IsoflavoneAnthocyanidin
genisteinhesperetinluteolinquercetin cyanidincatechin
sesamin
Diarylheptanoids
OH O
O
OH
O
O
OH
OH
OH
OH
OH
O OOH
Stilbenes Ellagitannins Coumarins
resveratrol ellagic acid coumarin
ferulic acid
A
B
C
OH

6. Polyphenol are rich in potential, however…
Limited bioactivity
Less than 5% of the
intake is absorbed and
reaches the plasma
unchanged
(Cao et al. 2015).
Limited application
Most of the products
are formulated into
capsules and tablets.
Low
solubility
Low
bioavailability
Hesperetin
Resveratrol
Apigenin
Naringenin
Isoflavone
Quercetin
Luteolin

7. The Problem –Bioavailability
7
Placebo Effect?
If a phytochemicals has a
bioavailability lower than 5%......

8. RenoSorb
TM
technique
Boosting Bioavailability
NATURALLY
•PATENTED
•SUPERIOR ABSORBTION
•TOTALLY WATER SOLUBLE
•WITHOUT EXCIPIENTS
•HIGH PURITY PRODUCTS
•ECO-FRIENDLY
•BIOLOGICAL PROCESS
Totally soluble in water

9. 9
Water barrier
(Unstirred water layer) System circulationIntestinal lumen
Native
polyphenol
P
P
Phosphorylated
pro-form Original form
Phosphate prodrug strategy
P
Alkaline phosphatase

10. Press me
I can move !

11. Advantages
High Purity
Low Purity
Water
Soluble
Water
Dispersion
Cyclodextrin-
based
technique
CWD
technique
Formulation
technique
Liposome
technique
CGSF
technique
SEDDS
technique
technique
Technology
~2000
Water
Insoluble
Technology
~2010
Water
Dispersion
RenoSorb
™
技術
2020
Totally
Water Soluble
11

13. The Cutting-Edge Technology
Our Innovation is Your Solution
Next Generation Nutraceuticals –soluble form for maximum efficacy
Nutraceutical
Pharmaceutical Skincare
Pet health

14. Product Pipelines
14
Research & Development Scale- up Production Product Launch

15. Product Pipelines
15
RenoFlavone
Fermented Soy Extract
RenoCidin
Fermented Citrus Bioflavonoids
RenoSage
Fermented Chrysanthemum Extract
Women
Health
Cardiovascular
Health
Healthy
Aging
Circulation
Health
Brain and
Neuro Health
Gout
Relieve
TM TM TM

16. EU.US.JP.KR.TW
No.
1283

17. 0 2 4 6
0
500
1000
1500
Time after oral administration (h)
Plasma genistein conc. (nM)
Native genistein
RenoFlaovne
TM
**
**
**
**
**
**
Pharmacokinetic Study of Genistein Phosphate
[Journal of Functional Foods13 (2015): 323- 335.]
Meanplasmaconcentration- timeprofilesof
genisteininratsafteroraladminstrationofnative
genisteinandRenoFlavone
TM
at3.7μmol/kgbw.
Dataaremean±SE(n=4).
**: p < 0.01 RenoFlavone
TM
vs Genistein
PKparameters RenoFlavone
TM
Genistein
AUC
0-∞ (μM・h) 2.0 0.5
C
max(μM) 1.3 0.1
T
max(h) 0.5 3.5
17
17

18. RenoFlavone
TM
next-generation genistein
•SCIENTIFICALLY PROVEN HEALTH
EFFECTS
•UNIQUE MODE OF ACTION
•LOW DAILY DOSE
•SUPERIOR BIOAVAILABILITY
•WATER SOLUBLE
•PATENT PROTECTED
Improve bioavailability
Improve overall
bioavailability to 370%
from original genistein.
Prevent post-
menopausal bone loss
About 500% better bone
minerals retained than
control.
Retain bone elasticity
Improve overall Young’s modulus about 140% from placebo group.
18

19. Interaction of Isoflavone With Estrogen Receptors
19
SERM
Selective estrogen-
receptor modulator
Ligand
Relativebinding affinity
β/α
hERα hERβ
Estradiol100 100 1
Genistein0.017 7.4 440
Daidzein 0.01 0.04 4
(±)Equol 0.2 1.6 8
Table. Relative binding affinity of compounds to hERβ/ α
(Muthyalaet al., 2004)
ER-αand ER-βreceptors predominate in various tissues
Brain
Heart/
arteries
Bone
Breast
Uterus
α
β

20. Placebo
Femur head
Cortical
Cortical
Femur head
Femur
bone
Micro CT image of
experimental rats
femur trabeculae at
12
th
week
Ovariectomized
Rat
X-Y Axis
X-Y Axis
Z Axis
Z Axis
Tracebularbone reference site: see
growth plate than offset 50 slice
(amount: 0.86926 mm), hight200
slice (amount: 3.4771 mm)
RenoFlavone
20

21. 21
RenoFlavone
Fermented Soy Extract
WOMENHEALTH
CARDIOVASCULAR HEALTH
21
TM
21

22. 22
Randomized, double-blind, placebo -control
A sample of 120 women (aged 18-40) suffering from primary
dysmenorrhea were randomly assigned to either a placebo
group or a RenoFlavone™group, and a 3-month human trial
was conducted.
Chi-Hao Wu, PhD
National Taiwan Normal University
Programs of Nutrition Science
Distinguished Associate Professor
Primary dysmenorrhea
Questionnaire Assessment :
•PSST (Premenstrual Symptoms Screening Tool)
•MDQ (Menstrual Distress Questionnaire)
•SF-MPQ (Short-Form McGill Pain Questionnaire)
•VAS (Visual Analogue Scale for Pain)
Genistein inhibits Ca
2+
-dependent uterine smooth muscle contraction and estrogen activity
and Alleviates Dysmenorrhea and Premenstrual Syndrome Symptoms
Human Trial of RenoFlavone
™
5 days before and after menstruation
(repeated 3 cycle)
RenoFlavone
™
40 mg
Premenstrual Syndrome
22

23. 23
The context of the beginning…
ClinicalTrial of RenoFlavone
™
[Cell185.10 (2022): 1676- 1693.]

24. ★10,000X Aqueous Solubility
★6X Superior Oral Bioavailability
★Fast Absorption
★Safety Studies
Native
hesperetin
Water-
soluble
Superior
Bioavailability
Scientific
Publication
Global
Patent
USFDA NDIN NO.1253
New Dietary Ingredient
O
O
OH
OH
O
OCH
3
P
OH
OH
O
Hesperetin phosphate
FDANotification
New Dietary
Ingredient
No.
1253

25. RenoCidin
TM
RenoCidin
TM
Hesperidin
Commercial
water soluble
hesperidin
Hesperidin
Commercial
water soluble
hesperidin
608%
3281%
Elevate the maximum
plasma level
Improve
bioavailability
RenoCidin
TM
is more bioavailable
than commercial hesperidin and
existed water soluble hesperidin
RenoCidin
TM
improved the
maximum plasma level of
hesperetinin systemic
circulation by about 33 times of
commercial products
•WATER SOLUBLE
•SUPERIOR BIOAVAILABILITY
•LOW DOSAGE REQUIRED
RenoCidin
TM
The next-generation hesperetin

26. Pharmacokinetic Study of HesperetinPhosphate
34
[Journal of Agricultural and Food Chemistry
69.35 (2021): 10184- 10193.]
Meanplasmaconcentration- timeprofilesof
hesperetininratsafteroraladminstration
ofnativehesperidinandRenoCidin
TM
at22.8
μmol/kgbw.Dataaremean±SE(n=4).
0 2 4 6
0
40
80
120
Time after oral administration (h)
Plasma hesperetin conc. (nM)
Native hesperidin
RenoCidin
TM
**
**
**
**
**
**
**
**: p < 0.01 RenoCidin
TM
vs Hesperidin
PKparameters RenoCidin
TM
Hesperidin
AUC
0-∞ (nM・h) 122.8 20.2
C
max(nM) 109.7 2.8
T
max(h) 0.25 4.0

27. RenoCidin
Fermented Citrus Bioflavonoids
METABOLIC WELLNESS
HEALTHY AGING
27
TM

28. 28Latest Reference: Hesperetinand longevity gene Cisd2
Results: Hesperetinactivates cisd2in mice, delaying ageing and promoting healthy longevity.
Cisd2 expression
in cardiac muscle
100% 45% 130%
Cardicfunction
Mechanical function
Electrical Function
Control Hesperetin
Young Ageing mice
對照組 橙皮素
27
30
33
週齡
(weeks)
Maximum
Lifespan
+ 13.8%
ControlHesperetin
Abdominal
Fat
對照組 橙皮素
0
2
4
6
體積
(mm
3
×10
3
)
-65%
ControlHesperetin
對照組 橙皮素
24
27
30
週齡 (weeks)
Average
Lifesapn
+ 8.7%
ControlHesperetin
Body Fat
對照組 橙皮素
0
2
4
6
8
10
體積
(mm
3
×10
3
)
-44%
ControlHesperetin
ControlHesperetin
microCTLean Tissue
HES
microCTBody Fat
ControlHesperetin
Hesperetin
Control
Survival (%)
Age (week old) Human
Age 85
Human
Age 96
Human age 78 Human age 84.5
Providing natural aging mice with Hesperetin(21
weeks old = equivalent to 70 years old in humans).
Hesperetin (100 mg/kg/day)
[Journal of Biomedical Science29.1 (2022): 53.]
Cisd2 expression
in skeletal muscle
100% 18% 150%
Sarcopenia
Muscle weight
Muscle strength
Control Hesperetin
Young Ageing mice
Volume (mm
3
x10
3
)
Volume (mm
3
x10
3
)
Age (Weeks) Age (Weeks)

29. 29Human Study of RenoCidin
™
Results: RenoCidin™ promoted peripheral vasodilation and enhanced blood circulation.
-5 0 5 10 15 20
12
16
20
24
Time after oral administration (h)
Skin temperature (
o
C)
PlaceboRenoCidin
TM
Time after cooling stress (min)
RenoCidin
™
Pre. 0 10 20
Control
[In-house data]
-Cooling Pressure Test: Ice water at 10°C for 5 min.
-Experimental Groups:
[Placebo] 200 mL of drinking water.
[RenoCidin™] 50 mg of RenoCidin ™dissolved in
200 mL of drinking water.
-Measurement Instrument: Thermal Imaging Camera.Condition: 25
o
C, RH 60% with solution at 25
o
C, n = 5.
Chen Hsu, PhD
R&D Director
Department of R&D
Hughes Biotech. Co., Ltd.
Evaluation of changes in skin temperature before and
after the cooling experiment to assess the
improvement in blood circulation effectiveness.
Cooling
stress
**
**
**
Value are mean ±SEM.**p< 0.01. RenoCidinvsplacebo group (paired t-test).
Suggested dose：50 mg/day

30. 30Human Trial of RenoCidin
™
-I
Chi-Chang Huang, PhD
Distinguished Professor
Graduate Institute of Sports Science
National Taiwan Sport University
Hyperlipidemia
Randomized, double-blind, placebo -control
Sample consists of 32 subjects with
borderline hypertriglyceridemia (TG >150),
randomized into placebo and RenoCidin™
groups, for an 8-week human trial.
Borderline Hypertriglyceridemia
TG: > 150 mg/dL (fasting)
Triglycerides (mg/dL)
Blood Metabolism Evaluation Indices
[Serum Biochemical Analysis]
•AST, ALT, BUN, creatinine, uric acid
•TG, TC, HDL-C, LDL-C
•Total Antioxidant Capacity
100 mg RenoCidin
™
/day, 8weeks
RenoCidin
™
Results: RenoCidin™ can reduce of blood lipid metabolism markers significantly in individuals
with borderline hyperlipidemia.

31. 31Human Trial of RenoCidin
™
-II
Randomized, double-blind, placebo-control
Sample consists of 32 healthy national athletes,
randAomizedinto placebo and RenoCidin™
groups, for an 8-week human trial.
Chi-Chang Huang, PhD
Distinguished Professor
Graduate Institute of Sports Science
National Taiwan Sport University
Evaluation indices for enhancing athletic performance:
[Biomechanical Testing]
•CMJ, Watt, IMTP, VO2max [Blood Biochemical
Analysis]
•Testosterone, Cortisol, hs-CRP, CK
Reduce fatigue
Enhance performance
Boost Athleticism
100 mg RenoCidin
™
/day, 8weeks
RenoCidin
™
Results: RenoCidin™ enhanced athletic performance and procidedanti-fatigue effects.

32. RenoSage
The next-generation luteolin
USFDA NDINapplication
NDIN No.1331 (pending)
Water-
soluble
Superior
Bioavailability
Scientific
Publication
Global
Patent
★84,000X Aqueous Solubility
★8.2X Superior Bioavailability
★Fast Absorption
★Safety Studies
Native
luteolin
RenoSage
TM
32
TM
Luteolin phosphate

33. •WATER SOLUBLE
•LOW DOSAGE REQUIRED
•SUPERIOR BIOAVAILABILITY
•100% FOODGRADE
COMPONENTS
RenoSage
TM
(next-generation luteolin)
Elevated the maximum
plasma level
Improved Bioavailability
RenoSage
TM
is more bioavailable
than commercial luteolin and
existed water soluble luteolin
RenoSage
TM
improved the
maximum plasma level of luteolin
in systemic circulation by about
21.5times of commercial products
1
1
0
2
4
6
AUC
0-∞
(µM ·h)
820 %
RenoSage
TM
1
1
0
1
2
3
C
m ax
(µM)
RenoSage
TM
Luteolin
2150 %
Luteolin

34. Pharmacokinetic Study of Luteolin Phosphate
37
[Unpublished data]
Meanplasmaconcentration-timeprofilesof
luteolininratsafteroraladminstration
ofnativehesperidinandRenoSage
TM
at175
μmol/kgbw.Dataaremean±SE(n=4).
0 2 4 6 8
0
1000
2000
3000
Time after oral administration (h)
Plasma luteolin conc. (nM)
**
**
**
**
**
**: p < 0.01 RenoSage
TM
vs Luteolin
PKparameters RenoSage
TM
Luteolin
AUC
0-∞ (μM・h) 5.58 0.68
C
max(μM) 3.01 0.14
T
max(h) 0.15 60

35. RenoSage
Fermented Chrysanthemum Extract
GOUTRELIEVE
BRAIN & NEURO HEALTH
35
TM

36. 36Latest Reference: Luteolin improved cognitive function
Results: Luteolin improved cognitive function in Alzheimer's disease mice by inhibiting
neuronal mitochondrial damage.
BrainLung LiverKidney
0.00
0.02
0.04
0.06
0.08
0.10
Organ luteolin con. (nmol/g)
8-week treatments (n = 10)
Luteolin 40 mg/kg (DMSO)
Tg-AD mice
Step-down
avoidance test
Open filed
test
Novel object
recognition test
Sacrifice
Triple transgenic Alzheimer's disease mice
3-day
interval
3-day
interval
3-day
interval
[Novel object recognition test]
ControlTg-ADTg-AD
Lut20
Tg-AD
Lut40
Attenuation of cognitive impairments by luteolin in Tg-AD mice. Illustration of novel object recognition
task. Assessing the effects of luteolin on recognition index (%) in testing stage after 24 h.#: WT group
vs. Tggroup; *: Tg+ Lutgroup vs. Tggroup. n=10 mice;
#
P < 0.05and *P < 0.05.
Tissue distribution of luteolin in rat at 4 h after oral administration.
Values are expressed as means + SD ofconcentrationof each
compound in targeted tissues (nmol/g) obtained from 5 rats.
[Redox Biology66 (2023): 102848.]
[Unpublished data]
Tissue distribution of luteolin in rats after
oral administration of RenoSage ™

37. 37Latest Reference: Luteolin improved cognitive function
Results: Luteolin effectively reduced uric acid levels in hyperuricemic mice by inhibiting
xanthine oxidase (XO) activity.
0
200
400
600
800
Uric acid in serum (mg/L)
**
##
Model LuteolinControl
-4.2X
Effects of luteolin on uric acid in serum of potassium oxonate(PO)-induced
hyperuricemia mice. Data represent mean ±SEM for 10 mice.Mice were
administrated with luteolin (100 mg/kg/day for 7 days, n=10) or treated with
only vehicle (saline 10 ml/kg/day, n=10) as model group after PO-induced
hyperuricemia in mice.
##
p< 0.01 compared with normal group
Substrates
Xanthine oxidase
IC
50(µM)
Allopurinol 0.25 ±0.01
Luteolin (LUT)
0.21 ±0.02
LUT 3’-O-sulfate 0.13 ±0.01
LUT 3’-O-GlcAc 3.22 ±0.24
LUT 7-O-GlcAc 5.20 ±0.38
Quercetin 11.02 ±0.51
Luteolin and its metabolites inhibit XO activity
The uric acid- lowering effect of luteolin in
hyperuricemicmice
IC50 values of allopurinol, luteolin and its
metaboliteson XO inhibition
Gout and Hyperuricemia
Joint Inflammation
Swelling
Uric Acid Crystals
Gouty Tophi
Hypoxanthine Xanthine Uric acid
O
2+H
2OH
2O
2O
2+H
2OH
2O
2
xanthine
oxidase
xanthine
oxidase
Purine metabolism. During the final stage of the purine metabolism, xanthine oxidase breaks down hypoxanthine to xanthine, and
xanthine to uric acid, while reactive oxygen species are formed.
These free radicals inactivate the vasodilator nitric oxide.
[Phytomedicine41 (2018): 54- 61.]
Concentration dependent inhibitory effects of LUT and its
metabolites and allopurinol on xanthine oxidation (substrate
concentration = 5 μM; n = 3). GlcAc: glucuronic acid.
[Biomedicine & Pharmacotherapy167 (2023): 115548.]

38. Patented and
scientifically
proven
Extensive
applications
Water soluble, easily absorbed
38
Fast absorption
Unique mechanism leads to
fast and efficientabsorption.
Real efficacy
Unique mechanism makes nutraceutical with real efficacy.
Take less, but get more
Better bioavailability means better efficient. Efficient absorption reduce placebowaste.
Better accessibility
Expanded diversity of consumer products, senior-friendliness.

39. Our Capabilities
Scientific
publications
GLP safety
studies
39
US,EU, JP, KR, TW
Patents
Certification

40. Efficacy‧Efficiency
Exciting New Levels“
“
HUGHES Biotech Teams

41. We make it better
Hughes Biotech
Innovative Nutraceutical Technology
Claire Fang
Director of Business Development
[email protected]
LinkedIn:clairefang-hughes