Glycative Stress Research
Online edition : ISSN 2188-3610
Print edition : ISSN 2188-3602
Received : December 31, 2019
Accepted : January 18, 2020
Published online : January 31, 2020
[Epub ahead of regular issues]
doi:10.24659/gsr.7.1_1

Original article

Safety and efficacy of nanoparticulated brown rice germ extract on reduction ofbody fat mass and improvement of fuel metabolism in both pre-obese and mildobese subjects without excess of visceral fat accumulation
Hiroaki Masuzaki 1), Kotaro Fukuda 2), Mika Ogata 2), Ayano Kinjo 2), Takeru Matsuo 2), Jun Nishihira 3)
1) Division of Endocrinology, Diabetes and Metabolism, Hematology and Rheumatology (Second Department of Internal Medicine),
Graduate School of Medicine, University of the Ryukyus, Nishihara, Okinawa, Japan
2) Sentan Pharma Inc., Fukuoka, Japan
3) Department of Medical Management and Informatics, Hokkaido Information University, Ebetsu, Hokkaido, Japan

Abstract

Beneficial effects on obesity and dysmetabolism of a variety of functional ingredients contained in brown rice haveattracted academic as well as clinical attention. On the other hand, it is noteworthy that the absorption efficiency of some of
the functional ingredients derived from brown rice from the digestive tract is significantly low. Based on these backgrounds,
we developed the “nanoparticulated brown rice germ extract” by encapsulating brown rice germ extract containing
γ-oryzanol, one of the functional ingredients abundantly and specifically in brown rice (rice bran), in nanoparticles, andconducted safety and efficacy tests in humans. The present study was performed appropriately after the approval from the
ethics committee of Hokkaido Information University. Japanese men and women whose BMI is 23 kg/m 2 or more and lessthan 25 kg/m2, or 25 kg/m2 or more and less than 30 kg/m2, and whose visceral fat area is less than 100 cm 2 when measured
by bioelectric-impedance methods were recruited for the study. We conducted a placebo-controlled, randomized, doubleblind, parallel-group comparative study targeting approximately 78 people who met the eligible criteria, and examinedmetabolic improvements when the "nanoparticulated brown rice germ extract" tablets were ingested once-a-day continuouslyfor 12 weeks. Although a decreasing trend was observed in the abdominal visceral fat area, which is a primary endpoint,
there were no significant differences among the ingestion groups of test food (low dose group: 5 mg, and medium-dose group:
10 mg). Among the secondary endpoints, plasma LDL-cholesterol (LDL-C) level showed a marginally significant decreasein the medium and low dose (5 mg) groups compared to the control group (placebo ingestion) after 8 weeks of ingestion( p = 0.078, p = 0.098). The plasma lipid ratio (LDL- C/ HDL- C) showed a significant decrease in the low dose groupcompared to the control group after 4 and 8 weeks of ingestion ( p = 0.016, p = 0.018). The plasma non-HDL- C level showeda marginally significant or significant decrease in the medium and low dose groups compared to the control group after 8
weeks of ingestion (p = 0.099, p = 0.031). Blood level of HbA1c also showed a significant decrease in the medium-dose groupcompared to the control group after 4, 8, and 12 weeks of ingestion (p = 0.029, p = 0.011, p = 0.011). Furthermore, plasma
level of adiponectin, which has potent anti-atherosclerotic actions, showed a marked increase in the high, medium, and low
dose groups (p = 0.012, p = 0.039, p = 0.017 ) after 4 weeks of ingestion compared to the control group, and also showed amarginally significant or significant increase after 8 and 12 weeks of ingestion. During the hospital visit, problematic changes
were not observed in blood pressure, pulse rate, general blood tests, liver and kidney functions, and urinalysis, and adverse
events suggesting a causal relationship with the test food were not observed. This clinical intervention study confirmed thesafety of ingestion of the “ nanoparticulated brown rice germ extract ”, which was also proven to improve the atherogenic
lipid profile, reduce HbA1c level, and increase plasma adiponectin concentration. Results obtained tempt us to expect that
nanoparticulated brown rice germ extract is well eligible for practical use.

KEY WORDS: brown rice, γ-oryzanol, nanoparticulated brown rice germ extract, obesity disease,		

metabolically-beneficial impact, adiponectin

Corresponding to: Hiroaki Masuzaki, MD, PhD.
Division of Endocrinology, Diabetes and Metabolism, Hematology, Rheumatology,
Department of Medicine, Graduate School of Medicine, University of the Ryukyus
Uehara 207, Nakagami-gun Nishihara-cho, Okinawa, 903- 0215 Japan

Tel: +81-98-895-1146 Fax: +81-98-895-1415 e-mail: hiroaki@med.u-ryukyu.ac.jp
Co-authors: Fukuda K, fukuda@sentaniryou.co.jp; Ogata M, ogata@sentaniryou.co.jp;
Kinjo A, kinjo@sentaniryou.co.jp; Matsuo T, matsuo@sentaniryou.co.jp;
Nishihira J, nishihira@do-johodai.ac.jp_ 1 _Glycative Stress Research 2020; 7 (1) : 1- 12(c) Society for Glycative Stress ResearchImpact of Nanoparticulated Brown Rice Germ Extract on Body Fat Massplacebo food used in the present study was obtained bymixing foods and food additives conforming to the standardsof the Food Sanitation Act, which are widely distributed andsold, and have been extensively used as raw material for food.
It was also manufactured in a factory with a well-establishedquality and hygiene management system, and safety for its
use as food was sufficiently ensured with the pre-shipping
inspection.

Introduction

Beneficial effects on obesity and dysmetabolism of avariety of functional ingredients contained in brown rice
have attracted academic as well as clinical attention 1). For
the past several years, we have focused on the mechanismof action of γ-oryzanol, one of the functional ingredients
present abundantly and specifically in brown rice among awide variety of grains, and have elucidated that γ-oryzanol
reduces the dependence on animal fat by acting on the
hypothalamus and reward system in the brain 2, 3), improves
glucose metabolism by working on the pancreatic endocrine
cells such as α and β cells to increase glucose-responsive
insulin secretion 4, 5), and further, improves the intestinal flora
imbalance associated with obesity in mice 6).
It should be noted that, due to the extreme lipid-soluble
properties, the absorption efficiency of various kinds offunctional ingredients in brown rice from the digestive tract
is significantly low 1, 7). In this context, we developed a newmaterial in which γ-oryzanol is encapsulated in PLGA (poly(DL-lactide-co-glycolide)) nanoparticles, and reported that
there is a remarkable improvement in glucose, and lipid
metabolism is observed at a dose as low as approximately
1/100th of the commonly administered γ-oryzanol whenthe material was orally administered to genetically obese
diabetic model, ob/ob mice 6). If similar effects could
be reproduced in humans, we expected that it could be
developed as an excellent dietary supplement that would help
prevent lifestyle-related metabolic diseases and progressionof obesity.
Based on this background, in the present study, wedeveloped the “nanoparticulated brown rice germ extract,”
by encapsulating brown rice germ extract containing highly
amounts of γ-oryzanol in nanoparticles, and conductedclinical trials for a placebo-controlled randomized, doubleblind, parallel-group comparative study to validate thesafety and efficacy in improving metabolism in humans.

Basic Information on the Test Food and Placebo FoodThree doses of nanoparticulated brown rice germ
extract (tablets), high-dose (containing 20 mg), medium-dose(containing 10 mg), and low-dose (5 mg) were prepared, andwhen examining the efficacy, such as the reduction of bodyfat mass and improvement in dysmetabolism, the study was
conducted by allocating one of the three doses to each of thesubject groups.
Hydroxypropyl cellulose, brown rice germ extract,
maltitol, and sodium bicarbonate are used as raw materials
for the nanoparticles; maltitol, sodium bicarbonate, starch,
crystalline cellulose, and citric acid are used for the tablet
excipients, and highly dispersed silicon dioxide and sucrose
fatty acid esters are used for the tablet lubricants. No ne of
the raw materials correspond to food allergens that requirelabeling, or for which labeling is recommended.
In the case of high dose, properties of nanoparticulated
brown rice germ extract (tablets) were: 8 mm, packed inaluminum pouches (2 tablets/pouch), containing one tabletof 200 mg x 2 tablets = 400 mg (containing nanoparticulated
brown rice germ extract 10 mg/tablet x 2 tablets), and werestored at room temperature avoiding direct sunlight, hightemperature, and humidity. The subjects had to ingest 2
tablets once daily in the morning (after breakfast) (in casethey forget, then they have to consume the tablets either after
lunch or dinner) with cold or hot water. The expiry date was
one year from the date of manufacture (Manufactured by
Kanehide Bio Co., Ltd., Okinawa, Japan).
Placebo food tablets were: 8 mm, packed in aluminum
pouches (2 tablets/pouch), containing one tablet of 200 mg
x 2 tablets = 400 mg (placebo tablets x 2 tablets), which werestored at room temperature avoiding direct sunlight, hightemperature, and humidity. The raw materials used weremaltitol, sodium bicarbonate, starch, crystalline cellulose,
and citric acid for the tablet excipient, and highly dispersed
silicon dioxide and sucrose fatty acid esters for the tablet
lubricant. None of the raw materials correspond to foodallergens that require labeling, or for which labeling is
recommended. The method of ingestion was the same as forthe test food. The expiry date was also one year from thedate of manufacture (Manufactured by Kanehide Bio Co.,
Ltd.).
The testing agency stored the test food appropriately,
and after the test ended, the remaining test food was
disposed appropriately with the responsibility of the testing
agency. The test food (3 doses) and placebo food werevisually indistinguishable, and each test food was assigned
an allocation identification code, which was printed on thecorresponding test food package. The assignment informationof the allocation identification codes was stored securely
by the allocation manager to prevent disclosure. Deliveryof the test food for 12 weeks was couriered to the subjects

Subjects and Methods

Summary of the Test Food and Placebo Food
Based on our previous studies in mice 6), we developed atest food in which brown rice germ extract, which is an active
ingredient, was encapsulated in nanoparticles to improve
the absorption rate in the digestive tract. The nanoparticles
used in the present study are manufactured as “edible
nanoparticles,” and the surface structure is made of HPC(hydroxypropyl cellulose), which is used for food worldwide.
Brown rice germ extract, which is an active ingredient, isencapsulated inside the nanoparticles, and rice bran oil,which has long been used as food by the Japanese, is used asa raw material for the brown rice germ extract.
The safety of the nanoparticulated brown rice germ
extract was confirmed with animal tests (single-dose oral
toxicity test, reverse mutation test, chromosomal aberration
test, micronucleus test, and repeated dose oral toxicity test,
respectively). Quality was maintained by manufacturing
in a factory with a well-established quality and hygiene
management system, and safety for its use as food was
sufficiently ensured with the pre-shipping inspection. The_ 2 _Glycative Stress Research

participation in the present study were selected as subjects.
The following selection criteria were used to recruit the
subjects: 1) Japanese men and women between the ages of 20
and 65, and 2) Subjects whose body mass index (BMI) andserum triglyceride level meets any of the criteria (1) to (3);(1) BMI of less than 25.0 kg/m 2 and serum triglyceride levelof less than 150 mg/dL, (2) BMI ranging from high-normal toobese class I ( 23.0 kg/m2 or more and less than 30.0 kg/m 2 )(3) Serum triglyceride level is normal to high (120 mg/dL or
more and less than 150 mg/dL), and subjects who did notmeet the following exclusion criteria were screened.
Exclusion criteria: 1) Subjects who were under physician's
advice, treatment and/or medication for obesity, dyslipidemia
or diabetes, 2) Subjects with familial hypercholesterolemia, 3)
Subjects with severe cerebrovascular diseases, heart diseases,
liver diseases, kidney diseases, gastrointestinal diseases,
or infectious diseases requiring notification, 4) Subjects
with a history of major gastrointestinal surgery, such as
gastrectomy, gastrointestinal suture, or intestinal resection,5) Subjects with significant abnormalities in blood pressure
and blood test, 6) Subjects with severe anemia, 7) Pre- andpost-menopausal women with substantial changes in physicalcondition, 8) Subjects who may be allergic to drugs or foods(especially from the grass family), 9) Subjects who regularly

before the start date of ingestion. All subjects were notified
to return the remaining test food when visiting the laboratoryfor examination after 4, 8 and 12 weeks of ingestion.

Safety Evaluation on the Excessive Ingestion of
Nanoparticulated Brown Rice Germ Extract
For safety evaluation, the subjects were given thrice
the highest recommended daily dose ( 20 mg) of thenanoparticulated brown rice germ extract that was planned
to be used for efficacy validation described below, and wasto be ingested daily for 4 weeks. An open-label, uncontrolled
study was conducted, initially planned with 30 subjects,
but finally, the analysis was performed for 24 subjects. The
subject profile of the safety confirmation test are shown in
Table 1. As the primary endpoint, the types, severity, andfrequency of occurrence of adverse effects were tabulated
to evaluate the safety of the test food. Type, severity, andfrequency of adverse events were evaluated as the secondary
endpoint.
Volunteers registered at the Health Information Science
Research Center, Hokkaido Information University, whofully understood the significance, contents, and purposeof this test and prior written consent was obtained forTable 1. Profile of subjects in safety confirmation test on the excessive ingestion of nanoparticulated brown rice germ extract.

Sex

Male subject (number of people)
Male subject proportion (%)

Age

Mean (age)NumberSD

StatureNumberMean (cm)SD

Body weight

NumberMean ( kg)SD

BMI

NumberMean ( kg/m 2 )SD

Body fat percentage

NumberMean (%)
SD

NumberAbdominal visceral fat area
(Impedance method)

Mean (cm 2 )ingestion rate

Mean (%)

SD

NumberSD

Control

HD group

3
15.00
20
49.9
10.3
20
158.33
9.35
20
61.25
8.85
20
24.33
1.61
20
32.98
4.80
20
87.20
19.31
20
99.23
1.35

3
15.00
20
50.9
7.4
20
158.47
6.61
20
60.73
5.49
20
24.16
1.26
20
33.76
5.94
20
86.85
13.60
20
98.75
2.90

p-value
1.000
0.714

0.955

0.825

0.712

0.653

0.948

0.862

MD group
4
20.00
20
52.7
8.3
20
157.64
7.76
20
60.87
6.20
20
24.46
1.43
20
33.24
5.68
20
90.40
22.30
20
99.46
0.82

p-value
1.000
0.351

0.801

0.876

0.788

0.877

0.630

0.883

LD group
4
20.00
20
51.5
7.8
20
158.63
7.99
20
61.63
7.08
20
24.44
1.32
20
32.88
5.30
20
83.70
18.54
20
96.73
14.36

p-value
1.000
0.584

0.912

0.880

0.823

0.948

0.562

0.201

Control (n = 20): Ingestion group of placebo food. HD group (n = 20): High dose of test food (20 mg). MD group (n = 20): Medium dose of test food (10 mg). LD
group (n = 20): Low dose of test food ( 5 mg). Independent t-tests were performed for anthropometric measurements (stature, weight, BMI, body fat percentage)
and abdominal visceral fat area. Fisher’s exact test was performed for gender. The Mann-Whitney U test was performed for the ingestion rate. Test food,
nanoparticulated brown rice germ extract; BMI, body mass index; SD, standard deviation.

_ 3 _Impact of Nanoparticulated Brown Rice Germ Extract on Body Fat Massuse medicines, health foods, and supplements (including
ingredients such as polyphenols such as catechin andquercetin, licorice extracts containing glabridin, chitoglucan,
and dietary fibers such as indigestible dextrin) that may affectbody weight and body fat mass, 10) Individuals who regularly
use medicines, health foods, and supplements that affectplasma lipids ( γ-oryzanol, eicosapentaenoic acid [EPA],
docosahexaenoic acid [DHA], medium-chain fatty acids,
plant sterols, sesamin, turmeric, polyphenols, indigestible
dextrin dietary fibers or other ingredients), 11) Subjects who
regularly use medicines, health foods, and supplements( including dietary fibers such as indigestible dextrin, andproducts containing ingredients such as polyphenol ) that
may affect blood glucose level, 12) Excessive smokers andalcohol users, and subjects with extremely irregular diets,
13) Women who had donated 400 mL blood within 16 weeks
or men who had given 400 mL blood within 12 weeks
before starting the ingestion. Also, subjects who have given
200 mL blood within 4 weeks or blood components within
2 weeks before the start of ingestion, 14) Women who are
pregnant, possibly pregnant, or lactating, 15) Subjects whoare currently participating in other clinical trials or haveparticipated within the past 4 weeks, 16) Subjects determined
to be ineligible by the principal investigator.
Excessive smokers are defined as those smoking 21 ormore cigarettes per day. Alcohol addicts are defined as those
who drink 20 g of pure alcohol per day (180 mL of sake,
500 mL of beer, 60 mL of whiskey double, 90 mL of shochu(a traditional Japanese distilled spirit) or 2 glasses of wine
(about 200 mL)) more than 6 days a week. After screening,
the principal investigator selected 30 individuals who weredeemed appropriate to participate in the present study, butultimately only 27 completed the study.

Japan) was used to measure blood pressure and pulse rate.
Body composition analyzer DC -320 (manufactured by
Tanita Corporation, Tokyo, Japan) was used to measure bodyweight, body fat percentage and BMI.

Overview of Efficacy Validation for the Ingestion of
Nanoparticulated Brown Rice Germ Extract
Volunteers registered at the Health Information Science
Research Center, Hokkaido Information University, who fully
understood the significance, contents, and purpose of thistest and prior written consent was obtained for participationin this study were targeted for the present study.
During the recruitment, subjects had to report following
selection criteria: 1) Japanese men or women between theages of 20 and 65, 2) BMI of 23 kg/m 2 or more and less than
25 kg/m2 or 3) BMI of 25 kg/m2 or more and less than 30 kg/
m2, and the visceral fat area, evaluated with the BEI method,is less than 100 cm2, and the following exclusion criteria, thatis, 1) Subjects who are under physician's advice, treatment
and/or medication for obesity, dyslipidemia or diabetes, 2)
Subjects judged to have familial hypercholesterolemia, 3)
Subjects with severe cerebrovascular diseases, heart diseases,
liver diseases, kidney diseases, gastrointestinal diseases, orinfectious diseases requiring notification, 4) Subjects using apacemaker or defibrillator, 5) Subjects with a history ofmajor gastrointestinal surgery, such as gastrectomy,
gastrointestinal suture, or intestinal resection, 6) Subjects
with significant abnormalities in blood pressure andblood test, 7) Subjects with severe anemia, 8) Pre- and postmenopausal women with significant changes in physicalcondition, 9) Subjects who may be allergic to drugs or
foods (especially from the grass family), 10) Subjects who
regularly use medicines, health foods, and supplements(including ingredients such as polyphenols such as catechinand quercetin, licorice extracts containing glabridin,
chitoglucan, and dietary fiber such as indigestible dextrin)that may affect body weight and body fat mass, 11) Subjects
who regularly use medicines, health foods, and supplementsthat may affect plasma lipid profile (γ-oryzanol, EPA, DHA,
medium-chain fatty acids, plant sterols, sesamin, turmeric,polyphenols, indigestible dextrin dietary fibers, or otheringredients), 12) Subjects who regularly use medicines,
health foods, and supplements (including dietary fiber suchas indigestible dextrin, and products containing ingredientssuch as polyphenol) that may affect plasma glucose level,13) Excessive smokers and alcohol users, and subjects withextremely irregular diets, 14) Women who had donated 400
mL blood within 16 weeks or men who had donated 400 mL
blood within 12 weeks before starting the ingestion. Also,
those who donated 200 mL blood within 4 weeks or blood
components within 2 weeks before the start of ingestion, 15)
Women who are pregnant, possibly pregnant or lactating,16) Subjects who are currently participating in other clinical
trials or have participated within the past 4 weeks, 17)
Subjects judged by the principal investigator to be ineligible.
Those who were not in conflict with the above 17 items andwere judged by the principal investigator to be appropriate
for participation in the present study were selected as
subjects. The definitions of excessive smokers and alcoholusers were the same as in safety evaluation.

Evaluation Items in Safety Evaluation
The main evaluation items for safety evaluation are as
follows. (1) Interview with physician, (2) Anthropometricmeasurement (height, weight, body fat percentage, BMI, andvisceral fat area by bioelectric impedance [BEI] method), (3)Vital check (blood pressure and pulse rate during hospital
visit), (4) Lifestyle questionnaire, (5) Dietary questionnaire,(6) Blood tests ( blood collection volume: Approx. 13 mL
for blood collection tubes: 3): General blood tests (whiteblood cell [ WBC], red blood cell [RBC], hemoglobin [Hb],hematocrit [Ht], platelet [Plt]), liver functions (aspartatetransaminase [AST], Alanine transaminase [ALT],
γ-glutamyl transpeptidase [γ-GTP], alkaline phosphatase[ALP], lactate dehydrogenase [LDH]), renal functions ( bloodurea nitrogen [BUN], creatinine [CRE], uric acid [UA]),lipid metabolism (total cholesterol [ TC ], low-densitylipoprotein-cholesterol [LDL-C], high-density-lipoproteincholesterol [ HDL- C ], triglyceride [TG] ), glucose
metabolism (fasting blood sugar (FBS), HbA1c).
Furthermore, on the first day of ingestion, after 2 and 4
weeks of ingestion, and 2 weeks after the end of ingestion,
all items excluding Lifestyle and Dietary questionnaires
were implemented among the above items. The waist
circumference was measured at the navel level in thestanding position with both feet aligned and arms down on
both sides. Digital automatic blood pressure monitor HEM7080IC (manufactured by OMRON Corporation, Kyoto,_ 4 _Glycative Stress Research

stored the documents or password-protected electronic media
related to the allocation in a place that can be locked. After
that, the personal information executive notified the subjectsof the date, time, and location of the clinical study based on
the assignment results.
The test food ingestion period is 12 weeks, with 2 tabletsof the test food or placebo food to be ingested once a day
after breakfast with water or hot water. The tests were
performed for each subject on the screening day, the ingestion
start date of test food, and 4, 8, and 12 weeks after ingestion,
respectively. A washout period of one week was set before
starting the ingestion of the test food. The subjects were asked
to record and submit daily changes in physical condition, useof medicines that they do not generally use and ingestion oftest food in a diary for 13 weeks from one week before thestart of ingestion up to the final examination 12 weeks afteringestion.
The examination items on the screening day were, (1)Interview with physician, (2) Anthropometric measurements(height, weight, body fat percentage, BMI, and waist
circumference), (3) Measurement of visceral fat area by
BEI method, (4) Vital check (blood pressure and pulse rateduring hospital visit), (5) Lifestyle questionnaire, (6) Dietary
questionnaire, and (7) Blood tests (blood collection volume:
Approx. 13 mL for blood collection tubes: 3): General blood
tests (WBC, RBC, Hb, Ht, and Plt), liver function (AST,

Study Design and Evaluation Items for
Efficacy Validation
The present study was set as a placebo-controlled
randomized, double-blind, parallel-group comparative study.
The planned test group was, 1) 20 subjects ingesting the test
food (nanoparticulated brown rice germ extract 20 mg) ata high dose (high-dose [HD] group), 2) 20 subjects in thetest food medium-dose (10 mg) group (medium-dose [MD]group), 3) 20 subjects in the test food low-dose (5 mg) group(low-dose [LD] group), and 4) Placebo food ingestion group
20 subjects (control group), but the final number of subjects
was 78 with 19 ~ 20 in each group.
An allocation manager from a third-party refers to
the subject list, randomizes the subjects with the stratified
permutation block method based on gender, age, BMI,
and abdominal visceral fat area (using the BEI method) as
stratification factors and assigns the subjects to four groups.
Two independent t-tests were performed for the age, physicalmeasurements (height, weight, BMI, and body fat percentage),
and abdominal visceral fat area among the 4 groups. Fisher's
exact test was performed for gender. When judged at asignificance level of 5% per test, a significant difference
was not observed between the groups in age, height, weight,
BMI, body fat percentage, and visceral fat area. The subject
profiles are shown in Table 2. The allocation manager strictly

Table 2. Profile of the subjects in the effective validation study on the ingestion of nanoparticulated brown rice germ extract.

Sex

Male subject (number of people)
Male subject proportion (%)

Age

Mean (age)NumberSD

StatureNumberMean (cm)SD

Body weight

NumberMean ( kg)SD

BMI

NumberMean ( kg/m 2 )SD

Body fat percentage

NumberMean (%)
SD

NumberAbdominal visceral fat area
(Impedance method)

Mean (cm 2 )ingestion rate

Mean (%)

SD

NumberSD

Control

HD group

2
10.53
19
50.16
10.52
19
157.53
8.88
19
60.59
8.58
19
24.32
1.65
19
33.60
4.03
19
84.84
16.62
19
0.99
0.01

2
10.53
19
51.16
7.56
19
157.64
5.63
19
60.23
5.15
19
24.22
1.27
19
34.49
5.07
19
86.79
13.97
19
0.99
0.02

p-value
1.000
0.738

0.962

0.875

0.827

0.551

0.698

0.817

MD group
4
20.00
20
52.65
8.33
20
157.64
7.76
20
60.87
6.20
20
24.46
1.43
20
33.24
5.68
20
90.40
22.30
20
0.99
0.01

p-value
0.661
0.416

0.968

0.909

0.780

0.822

0.385

0.923

LD group
4
21.05
19
51.79
7.85
19
158.72
8.20
19
61.73
7.26
19
24.44
1.36
19
32.93
5.44
19
84.58
18.62
19
1.00
0.00

p-value
0.66
0.591

0.669

0.660

0.807

0.667

0.964

0.163

Control (n = 20): Ingestion group of placebo food. HD group (n = 20): High dose of test food (20 mg). MD group (n = 20): Medium dose of test food (10 mg). LD
group (n = 20): Low dose of test food (5 mg). Independent t-tests were performed for anthropometric measurements (stature, weight, BMI, body fat percentage)
and abdominal visceral fat area. Fisher’s exact test was performed for gender. The Mann-Whitney U test was performed for the ingestion rate. Test food,
nanoparticulated brown rice germ extract; BMI, body mass index; SD, standard deviation.

_ 5 _Impact of Nanoparticulated Brown Rice Germ Extract on Body Fat Mass

- Protection of subject rights and acquisition of

ALT, γ- GTP, ALP, and LDH), renal functions (BUN, CRE,
and UA), lipid metabolism profile (TC, LDL-C, HDL- C,
and TG), and glucose metabolism (FBS, HbA1c).
The following were performed on ingestion start date,
and after 4, 8, and 12 weeks of ingestion: (1) Interview
with physician, (2) Anthropometric measurements (weight,
body fat percentage, BMI, and waist circumference), (3)Measurement of visceral fat area by BEI method (notperformed after 4 weeks) (4) Vital check (blood pressure
and pulse rate during hospital visit), (5) Personality
questionnaire (Japanese version TIPI-J) (only on ingestion
start date), and (6) Blood tests ( blood collection volume:
Approx. 13 mL, blood collection tubes: 3): General blood
tests (WBC, RBC, Hb, Ht, and Plt), liver function (AST,
ALT, γ-GTP, ALP, and LDH), renal functions (BUN, CRE,
and UA), lipid metabolism profile ( TC, LDL-C, HDL- C,
TG, LDL-C/HDL-C, and non-HDL), glucose metabolism(FBS, HbA1c, insulin, and homeostasis model assessment
for insulin resistance [HOMA-IR]), adipokines ( bloodlevels of adiponectin and leptin), urinalysis (volume ofurine collected: approx. 10 mL, urine sample tube 1 No.), andurinalysis (pH, protein, sugar, urobilinogen, bilirubin, ketone
bodies, and occult blood).

informed consent for safety evaluation

The present study was conducted after the deliberationby the Ethics Committee of Hokkaido Information
University, and before starting the study, the following content
was explained to the subjects, and informed consent forparticipation in the present study was obtained in writing.
That is, 1) Name of the study, Ethics Committee has reviewed
and the Committee President has approved the study, 2)Name, affiliation and contact information of the principal
investigator, consultation office for the tests and medical
institutions conducting the tests, 3) The purpose and methodof the study, 4) Expected benefits and potential disadvantagesof participating in the study, 5) Subjects can withdraw fromthe study at any time, 6) Subjects will not be adversely
affected by not participating in or withdrawing from thestudy, 7) Methods for anonymizing, storing, and disposingof samples and information to maintain confidentiality ofthe subjects, 8) Matters concerning compensation for healthhazards requiring medical treatment, 9) Matters relating to
the cost of the study, 10) Matters concerning the researchfunding and conflicts of interest, and 11) Methods withwhich the study information will be disclosed. We respected
the protection of the privacy of the subjects and stored theinformation with the strict compliance.

Statistical Analysis- Handling of data and storage of records during

The result was indicated as the mean value ± standard
deviation. For the safety test, 2 independent t -tests were
performed for age and physical measurements (height,
weight, BMI, and body fat percentage). Fisher's exact test was
performed for gender. For the efficacy test, 2 independentt-tests were performed for the age, physical measurements(height, weight, BMI, and body fat percentage), andabdominal visceral fat area, and a Fisher's exact test was
performed for gender. A Mann-Whitney U test was performed
for comparison between groups. The abdominal visceral fat
area was evaluated by performing 2 independent t-tests forthe change in the amount from before the start of ingestion tothat at each time point of evaluation. The significance level inthe statistical hypothesis test was 5% with the two-sided test.

safety evaluation

When handling data of subjects and materials obtained
during the present study, the personal information executive
will carefully anonymize the information to protect theprivacy of the subjects so that just the information cannot
be used to identify individuals. The information wasanonymized by deleting personal information that could
identify individual subjects and assigning an anonymous
number not related to personal information. The tables
relating the personal information and the anonymous number
was properly managed and kept at a location that can belocked under the direction of the personal information
manager. When information is stored as electronic data, apassword is set for the electronic media and kept at a locationthat can be locked.
The data and materials will be retained for 5 years
after the completion of the study or 3 years after the final
publication of results, whichever is later. Even when thematerials are disposed of, personal information will be strictly
anonymized to prevent disclosure of personal information.
Subject data obtained in this test was anonymized to protect
personal information before using the data for comparisonand correlation analysis with other studies, publication in
papers, presenting at academic conferences, management ofregistered volunteers, and quality improvement of subsequentstudies.

Ethical Guidelines- Benefits and measures to minimize the disadvantage
and risks of a subject during safety evaluation

The benefits to the subjects are that regular blood tests
will be conducted and results can be immediately obtained
from the physicians. The disadvantages include, test foodhas to be ingested daily, time and effort are required tomake entries in the diary, regular visits to the hospital cause
time constraints, and blood collections and various tests
cause a burden. As measures to minimize risks, safety was
further ensured with the exclusion criteria, and we worked incooperation with the medical institutions conducting the tests
to prevent health hazards during the onset of adverse events.
For ensuring that subjects obtain medical examination whenrequired due to poor physical conditions, the name of the
medical institution and name of the concerned physician will
be communicated to the consultation office, and a system isestablished so that the principal investigator can promptly
take appropriate measures when necessary.

- Ethical review

The present study was conducted in compliance with
the Ethical Guidelines for Medical and Health Research
Involving Human Subjects (February 28, 2017, partiallyrevised by the Ministry of Education, Culture, Sports,
Science, and Technology and Ministry of Health, Labour,and Welfare) and the Helsinki Declaration (amended at the
WMA General Assembly in Fortaleza held in October 2013).

_ 6 _Glycative Stress Research

We obtained the approval for the above ethics review to carryout the study.
Safety Evaluation for Excessive Ingestion of
Nanoparticulated Brown Rice Germ Extract
Ethics Review Number: 2017-22 (approved on October30, 2017), 2017-23 (change approved on November 24, 2017)
in the ethics committee of Hokkaido Information University.
Dose Determination Study of Nanoparticulated Brown Rice
Germ Extract on Body Fat Reduction by Daily Ingestion
Ethics Review Number: 2017-18 (approved on September
25, 2017), 2017-19 (approved on October 27, 2017).

with normal BMI and TG showed a marginally significant
decrease 2 weeks after the end of ingestion compared tobefore starting the ingestion (p = 0.084). Meanwhile, thesubject group ranging from high-normal BMI to obese class I
showed a marginally significant or significant decrease after
4 weeks of ingestion and 2 weeks after the end of ingestion
compared to before starting ingestion (p = 0.057, p = 0.022,
respectively).
Regarding blood lipids profile, both TC and LDL -C
levels showed a significant decrease at every point in timecompared to the day ingestion was started. TC values
showed a significant reduction after 2 and 4 weeks ofingestion, and 2 weeks after the end of ingestion compared
to before starting ingestion ( p = 0.001, p = 0.001, p = 0.037,
respectively). Stratified analysis also showed a significant
decrease after 2 weeks of ingestion compared to before
starting ingestion for the subject group with normal BMI
and TG ( p = 0.029). Furthermore, the subject group ranging
from high-normal BMI to obese class I showed a significant
or marginally significant decrease after 2 and 4 weeks ofingestion, and 2 weeks after the end of ingestion compared tobefore starting the ingestion ( p = 0.033, p = 0.001, p = 0.098,
respectively). LDL- C values showed a significant decrease
after 2 and 4 weeks of ingestion, and 2 weeks after the endof ingestion compared to before starting the ingestion (p= 0.003, p = 0.009, p = 0.002). Stratified analysis showed amarginally significant decrease after 2 weeks of ingestion
compared to before starting ingestion for the subject groupwith normal BMI and TG (p = 0.082). Notably, the subject
group ranging from high-normal BMI to obese class I showeda significant decrease after 2 and 4 weeks of ingestion,and 2 weeks after the end of ingestion compared to before
starting the ingestion (p = 0.029, p = 0.004, p = 0.011,
respectively). On the other hand, no significant differences
or marginal significant differences were observed at any timein plasma HDL- C and TG levels.
Importantly, fasting blood glucose level in the earlymorning showed a significant decrease after 2 and 4 weeks
of ingestion, and 2 weeks after the end of ingestion compared
to before starting ingestion ( p = 0.001, p = 0.026, p = 0.001,
respectively). In the stratified analysis, the subject group
ranging from high-normal BMI to obese class I showed asignificant decrease after 2 and 4 weeks of ingestion, and 2
weeks after the end of ingestion compared to before starting
the ingestion (p = 0.001, p = 0.010, p = 0.004, respectively).
The subject high-normal TG group showed a significant
decrease after 2 weeks of ingestion, and 2 weeks after theend of ingestion compared to before starting the ingestion(p = 0.029, p = 0.029). However, the blood HbA1c level
unexpectedly showed a significant increase after 2 weeks
of ingestion and 2 weeks after the end of the ingestion
compared to before starting the ingestion ( p < 0.001, p <0.001, respectively).
In humans and rodents, it has been reported thatγ-oryzanol contained in brown rice germ extract exemplifies
a line of metabolically-beneficial impact such as antiobesity, lipid-lowering, and glucose-lowering effects 1- 6). In
agreement with these notions, the present “safety evaluation”
study also suggests the test food may exert anti-obesity,
lipid-lowering, and glucose-lowering effects, especially insubjects with high BMI.

- Pre-registration

The UMIN test ID for the excessive ingestion of thetest food is UMIN000030323, Receipt No.: R000034628,
Test name: Safety Evaluation for Excessive Ingestion of
Nanoparticulated Brown Rice Germ Extract, Date ofdisclosure of the study information: December 8, 2017;
modified on July 20, 2018. The UMIN test ID for efficacy
validation is UMIN000029511, Receipt No.: R000033721,
Test name: Dose Determination Study of Nanoparticulated
Brown Rice Germ Extract on Body Fat Reduction by Daily
Ingestion, Date of disclosure of the study information:
October 13, 2017; modified on July 20, 2018.

Results

Safety Evaluation on the Excessive Ingestion of
Nanoparticulated Brown Rice Germ Extract

An open-label, uncontrolled study was conducted,
initially planned with 30 subjects, but finally, the analysis
was performed for 27 subjects. As the primary endpoint,
the types, severity, and frequency of occurrence of adverse
effects were tabulated to evaluate the safety of the test food.
The result of tabulating and evaluating the types, severity,and frequency of occurrence of adverse events as secondary
endpoint, showed that no noticeable adverse effects or
clinically problematic adverse events occurred for any of
the study items described under the section “Subjects andMethod, ” even if thrice the recommended daily intake fornanoparticulated brown rice germ extract is continued to beingested for 4 weeks. Of the initial 30 subjects, 2 subjects
were excluded because they were unable to ingest the test
food properly, while one subject accidentally happened todevelop the symptoms of the influenza virus during the study
period; thus, the study was stopped for the subject. Though
the subject was excluded, it was determined that there wasno causal relationship with the ingestion of the test food.
From the above, it was confirmed that there was no problem
in the safety of this test food within the framework of theevaluation system of daily ingestion for at least 4 weeks.
Regarding the body composition, a significant decreasein body weight, body fat mass, and BMI was observed atevery point in time compared to the day the ingestion wasstarted. In particular, the body fat percentage showed asignificant decrease after 4 weeks of ingestion and 2 weeks
after the end of ingestion ( p = 0.014, p = 0.017, respectively).
O f note, as a result of a stratified analysis of body fat mass,
the decrease in body fat mass was more noticeable in thesubject group with higher BMI. In detail, the subject group_ 7 _Impact of Nanoparticulated Brown Rice Germ Extract on Body Fat Mass

- Efficacy Validation for the Ingestion of
Nanoparticulated Brown Rice Germ Extractgroup (p = 0.028, Fig. 4-a) after 8 weeks of ingestion and
HD and LD groups showed a marginally significant increaseor significant increase compared to the control group (p =0.064, p = 0.021, respectively, Fig. 4-c) after 12 weeks ofingestion.
Body composition (weight, BMI, body fat percentage),vital signs (blood pressure and pulse rate during hospital
visit), general blood testing (WBC, RBC, Hb, Ht, and Plt),liver functions (AST, ALT, γ-GTP, ALP, and LDH), renal
functions (BUN, CRE, and UA) and urinalyses (protein,sugar, urobilinogen, bilirubin, ketone bodies, and occult
blood) did not show clinically problematic findings andadverse events having a causal relationship with the test foodwere not observed at all. From a line of results above, it was
concluded that there were no particular safety issues in thepresent “efficacy validation” study.

The abdominal visceral fat area before the start ofingestion, after 8 and 12 weeks of ingestion was 82.1 ±18.0 kg, 89.1 ± 24.1 kg, 82.2 ± 23.1 kg in the control group,and 86.8 ± 14.8 kg, 90.9 ± 18.4 kg in the HD group, 90.1
± 15.3 kg, 90.4 ± 23.3 kg, 89.4 ± 22.6 kg, 87.3 ± 23.0 kg in
the MD group, 86.0 ± 24.6 kg, 90.1 ± 22.9 kg, 84.2 ± 19.7
kg in the LD group. The amount of change from before thestart of ingestion to after 4 weeks of ingestion, the amountof change from before the start of ingestion to after 8 weeks
of ingestion, the amount of change from before the start ofingestion to after 12 weeks of ingestion, respectively, were
6.6 ± 16.0 kg and 0.1 ± 16.1 kg in the control group, 4.5 ±18.3 kg and 3.3 ± 17.6 kg in the HD group, 0.0 ± 19.8 kg and− 3.1 ± 10.9 kg in the MD group, and 4.1 ± 14.4 kg and −1.8
± 12.1 kg in the LD group.
As a result of the statistical analysis, a significant
increase is observed in the percentage of body fat of thecontrol group after 8 weeks of ingestion as compared tothe day of the screening ( p < 0.001). On the other hand,HD group (1), MD group (2), and LD group (3) somehow
showed slight but a significant increase after 8 and 12 weeks
of ingestion as compared to the day of screening ((1): p =0.026, p = 0.007, (2): p = 0.057, p = 0.054, (3) p = 0.021, p =0.050). Consequently, the abdominal visceral fat area, whichis the primary endpoint, showed a slight decrease in the test
food intake group compared to the control group, but dosedependence or a statistically significant difference was notobserved (Fig. 1 ). Significant differences and marginallysignificant differences were not observed among groups at
any time.
Regarding the secondary endpoints, such as body weightand BMI, significant differences, marginally significant
differences, and dose-dependence were not observed amonggroups at any time (Fig. 2). After 8 weeks of ingestion,
the plasma LDL-C level showed a marginally significant
decrease in the MD and LD groups compared to the control
group ( p = 0.078, p = 0.098), but a significant difference
and dose-dependency of the effects were not observed.
Significant differences and marginally significant differenceswere not observed among groups at any time for HDL-C.
The LDL-C/HDL-C ratio showed a significant decrease in
the LD group compared to the control group after 4 and 8
weeks of ingestion (p = 0.016, p = 0.018, respectively). After
8 weeks of ingestion, the serum non-HDL-C level, which isan indicator of atherogenic lipids, showed a trend of decreaseor a significant decrease in the MD and LD groups comparedto the control group ( p = 0.099, p = 0.031, respectively).
Significant differences and marginally significant
differences were not observed among the groups at any
point of time in the fasting blood glucose level, and dosedependency was also not observed ( Fig. 3-a). In contrast,
blood HbA1c levels were significantly reduced in the MD
group compared to the control group after 4, 8, and 12 weeks
of ingestion ( p = 0.029, p = 0.011, p = 0.011, respectively, Fig.
3-b ).
Noticeably, plasma concentration of adiponectin showeda significant increase in the HD, MD, and LD groups
compared to the control group after 4 weeks of ingestion (p= 0.012, p = 0.039, p = 0.017, respectively, Fig. 4-a). The HD
group showed a significant increase compared to the control

Discussion
In Japan, the proportion of obese people due toovernutrition and lack of physical exercise has increased inrecent years. According to the reports in the 2015 National
Health and Nutrition Survey by the Ministry of Health,
Labor, and Welfare, 29.5% of men and 19.2% of women
have been reported to fall in the obese category with a BMI
of 25 kg/m2 or more 8). Because obesity is closely associatedwith the development of a series of lifestyle-related diseases
such as type 2 diabetes, dyslipidemia, hypertension, a varietyof arteriosclerotic diseases, and non-alcoholic fatty liverdisease (NAFLD), prevention and amelioration of obesity
disease are important challenges worldwide.
Based on this background, there is a growing interestin foods that can be ingested daily and easily as well as
are effective in preventing and improving obesity disease
without causing adverse effects. Of note, a wide variety of
impact on metabolic of food ingredients such as brown rice
germ extract (γ-oryzanol) have been intensively reported
in recent years 1, 7). Brown rice is unpolished rice that isrich in functional ingredients such as vitamins, minerals,
dietary fibers, and γ-oryzanol, compared to white rice 1). In
particular, γ-oryzanol, which is specifically and abundantly
in brown rice, is an ester mixture of ferulic acid and several
kinds of phytosterols. In addition to its previously known
beneficial effects for dyslipidemia and dysregulation ofautonomic nervous system 1), our recent research in mice has
elucidated that γ-oryzanol reduces the dependence on animal
fat by acting on the hypothalamus and reward system inthe brain 2, 3), improves glucose metabolism by working on
pancreatic endocrine cells including α and β cells toincrease glucose-responsive insulin secretion 4, 5), and further,
improves the imbalance of gut microbiota associated withchronic feeding of high fat diet and obesity 6). Furthermore,in a crossover interventional study targeting 30- to 60-yearold men with metabolic syndrome living in Okinawa, thestaple food (white rice) was replaced by brown rice withthe same amount of calories in one meal daily withoutchanging the side dish menu, and consequently, postprandial hyperglycemia and obesity-associated metabolic
derangement were considerably improved (Brown Rice andVisceral Fat Obesity in Okinawa (BRAVO Study) 7). However,_ 8 _Figure 1
Glycative Stress Research

Figure 2

Fig. 1. Changes in abdominal visceral fat area 8 weeks after the ingestion of nanoparticulated brown rice
germ extract in each dose group.

Results are expressed as mean ± SE. Control (n = 20): Ingestion group of placebo food. HD group (n = 20): High dose of
test food (20 mg). MD group (n = 20): Medium dose of test food (10 mg). LD group (n = 20): Low dose of test food (5 mg).
Test food, nanoparticulated brown rice germ extract; SE, standard error.

a)

b)

a)

b)
b)

Fig. 2. Changes in body weight and BMI 4 weeks after the ingestion of nanoparticulated brown rice germ
extract in each dose group.

a) Body weight. b) BMI. Results are expressed as mean ± SE. Control (n = 20): Ingestion group of placebo food. HD group(n = 20): High dose of test food (20 mg). MD group (n = 20): Medium dose of test food (10 mg). LD group (n = 20): Low
dose of test food (5 mg). Test food, nanoparticulated brown rice germ extract; BMI, body mass index; SE, standard error.

_ 9 _Figure33
Figurea)

Impact of Nanoparticulated Brown Rice Germ Extract on Body Fat Massb) b)b)a)a)Fig. 3. Changes in FBS level and HbA1c level 4 weeks after the ingestion of nanoparticulated brown rice germ extract ineach dose group.
a) FBS. b) HbA1c. Results are expressed as mean ± SE. *p < 0.05, by t-test. Control (n = 20): Ingestion group of placebo food. HD group (n= 20): High dose of test food (20 mg). MD group (n = 20): Medium dose of test food (10 mg). LD group (n = 20): Low dose of test food (5mg). Test food, nanoparticulated brown rice germ extract; FBS, fasting blood glucose; SE, standard error.

Figure 4

a)
a)

b)b)

c)
c)

Fig. 4. Adiponectin changes after the ingestion of nanoparticulated brown rice germ in each dose group.

a) 4 weeks. b) 8 weeks. c) 12 weeks. Results are expressed as mean ± SE. *p < 0.05, by t test. Control (n = 20): Ingestion group of placebo
food. HD group (n = 20): High dose of test food (20 mg). MD group (n = 20): Medium dose of test food (10 mg). LD group (n = 20): Low
dose of test food (5 mg). Test food, nanoparticulated brown rice germ extract; SE, standard error.

_ 10 _Glycative Stress Research

there has long been concern that the absorption efficiency of
extremely-lipophilic γ-oryzanol is markedly diminished 1, 7).
It is therefore considered essential to devise innovative ways
to enable γ-oryzanol to be more efficiently absorbed byintestinal tract in human clinics.
Based on this background, we developed a new material
in which γ-oryzanol is encapsulated in PLGA nanoparticles,
and reported that there was a remarkable improvement indysmetabolism of glucose and lipids at a dose as low as about
1/100 th of the commonly administered dose of γ-oryzanol
when the material was orally administered to geneticallyobese diabetic ob/ob mice 6). In this context, aiming forthe application in humans, we developed the brand-new“nanoparticulated brown rice germ extract ” in which brownrice germ extract is encapsulated in nanoparticles. From theresults of oral administration of nanoparticulated brown rice
germ extract in mice, it was elucidated that the absorption
rate was exaggerated by approximately 5 times compared
to non-nanoparticulated brown rice germ extract. If similareffects would be reproduced in humans, we expect that it
could be developed as an excellent dietary supplement thatwould help prevent lifestyle-related metabolic diseases.
The safety evaluation of the present study showedthat no noticeable adverse effects or clinically problematic
adverse events occurred for any of the study items describedunder the section “ Subjects and Method,” even when thrice
the recommended daily intake for nanoparticulated brownrice germ extract is continued to be ingested for 4 weeks. It
should be noted that the nanoparticles used in the present
study are manufactured as “edible nanoparticles”, and thesurface structure is made of HP C (hydroxypropyl cellulose),which is widely used for food. On the other hand, the PLGA
nanoparticles used in our mouse experiments have not been
approved to be used as food additives for humans, and in thepresent human trial, this point was technically improved,
and the present study was conducted with utmost attention tosafety.
As described in the section “ Results ”, all the adverse
events in clinical findings were mild, and none of the cases
were clinically problematic, and the causal relationship withthe test food was determined to be negative as the cases
recovered or became less severe during the ingestion periodof the test food. The adverse events (abnormal fluctuations)in the laboratory evidence were considered to be within
the physiological range of fluctuations in daily life, and thecausal relationship was determined to be negative. From thenotion above, it was confirmed that there was no particular
problem in the safety of this test food within the frameworkof the evaluation system of daily ingestion for at least 4
weeks.
The efficacy validation test of the ingestion of thenanoparticulated brown rice germ extract, showed a slightdecrease in the abdominal visceral fat area, which is theprimary endpoint, in the test food group compared to thecontrol group, but regrettably, dose-dependency of the effects
or statistically-significant difference were not observed(Fig. 1 ). Among the secondary endpoints of body weightand BMI, significant differences, marginally significant
differences, and dose-dependency were not also observed
among groups at any time (Fig. 2). The plasma LDL-C level
showed a marginally significant decrease in the test foodgroup compared to the control group, however, no significant

difference and dose-dependency were observed.
On the other hand, the LDL-C/HDL-C ratio, a clinicalmarker for atherogenic lipid profile, showed a significant
decrease in the LD group compared to the control group. In
the same way, after 8 weeks of ingestion, the plasma nonHDL-C level, which is also an indicator for atherosclerotic
lipids, showed a trend to decrease or a significant decrease in
the MD and HD groups compared to the control group.
Significant differences and marginally significant
differences were not observed among the groups at any
point in time in the fasting blood glucose level, and dosedependency was also not observed. On the other hand, theblood HbA1c level after 4, 8, and 12 weeks of ingestion
showed a significant decrease in the MD group compared tothe control group. Notably, plasma adiponectin concentration
showed a significant increase in the HD, MD, and LD groups
compared to the control group after 4 weeks of ingestion.
Adiponectin, a potent anti-arteriosclerotic humoral mediator,
is a physiologically active substance and specifically secreted
from adipocyte 9). Since plasma adiponectin concentration
is well known to decrease sensitively by reflecting thedysfunction and accumulation of visceral fat 9), it is reasonable
to speculate that an apparent increase in the plasma
adiponectin concentration by ingestion of the nanoparticulated
brown rice germ extract may indicate an improvement invisceral fat function before the actual decrease in the visceralfat mass.
During the hospital visit by the subjects, problematicchanges were not observed in blood pressure and pulse
rate, general blood tests, liver and kidney functions, andurinalyses, and no adverse events having a causal relationshipwith the test food were observed. From these observations,the safety of daily ingestion of the “nanoparticulated brownrice germ extract ” for 12 weeks was confirmed in the present
study. Functionally, results obtained tempt us to expect thatthe extract may contribute, at least partly, to prevent theoccurrence or progression of arteriosclerotic and metabolic
diseases by improving the atherogenic lipid profile, reducing
the HbA1c levels and increasing the plasma adiponectinconcentration. However, a clear correlation between the doseand effect was not necessarily observed in the present study.
In this context, the determination of an appropriate dose
remains to be a challenge for future studies. The reason forsuch ambiguities of the results may also be that the numberof subjects for each dose was few at 19 ~ 20, due to whicha statistically significant difference could not be detected,
and further, the 12-week study period may have been too
short to represent a significant improvement in various
parameters for subjects without apparent metabolic orarteriosclerotic diseases. Since beneficial effects for obesity,
dyslipidemia, and hyperglycemia were pronounced insubjects having higher BMI in the safety evaluation study inwhich as much as thrice the middle dose (10 mg) used during
efficacy validation was ingested daily for 4 weeks, detailed
examination of the adequate ingestion amount by clinicalstudies is warranted in the next step.
Based on results of our basic and clinical research, aswell as those from other studies worldwide, brown rice canbe expected not only to reduce body fat mass and ameliorate
fuel metabolism but to improve cognitive dysfunction, aseries of addictive behaviors and to exert anti-aging impact 10).
We named the nanoparticulated brown rice germ extract_ 11 _

Impact of Nanoparticulated Brown Rice Germ Extract on Body Fat Mass

used in the present study as Brown Rice Foodicle® (Foodicle
is a combination of Food and Particle, which is the registeredtrademark of Sentan Pharma Co., Ltd). We are currently
developing materials for practical use that can bring out
a promising potential for a variety of health promotion ofbrown rice.

Conclusion
This clinical intervention study confirmed the safetyof the ingestion of the “nanoparticulated brown rice germ
extract,” which was also proven to improve the atherogenic
lipid profile, reduce HbA1c level, and increase plasma
adiponectin concentration. Results obtained tempt us to
expect that nanoparticulated brown rice germ extract is welleligible for practical use.

Conflict of Interest
This research received support from Sentan Pharma Inc.

Acknowledgement
The present study was supported in part by NEDO (New
Energy and Industrial Technology including Development
Organization, Innovative Manufacturing Industry Program),
2015, Strategic Basic Technology Upgrading Support
Program, 2016 and Strategic Basic Technology Upgrading
Support Program, 2017, respectively.

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