2. The sweetener according to claim 1, wherein the compound comprises one or more kinds of amino acid oi: peptide selected from the group consisting of y-Glu-X-Gly where X represents an amino acid or an amino acid derivative, y-Glu-Val-Y where Y represents an amino acid or an amino acid derivative, y-Glu-Ala, y-Giu-Gly, y-Glu-Cys, Y-Glu-Met, Y-Glu-Thr, y-Glu-Val, y-Glu-Orn, Asp-Gly, Cys-Gly, Cys-Met, Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, y-Glu-Met(0), Y-Glu-Y-Glu-Val, Y-Glu-Val-NH2, y-Glu-Val-o1, y-Glu-Ser, y-^lu-Tau, Y-(M ii-Cya(S-Me) (0) , Y~Gl'J~T-'eu, Y-Glu-ll<", y-'»'^'~*^-''"'> ^^'^id y~Glu~Cya(S-Me). J. 'I'ho HweeUonot: according to claim 2, whoroln X reproHonts CyHliiNO), Cys (S-al lyl) , Gly, Cys(S-Me), Abu, t.-],ou, CIB, Alb, Pen cii Htif, and Y represents Gly, Val, Glu, LyH, l'lu», Moi , Pro, Arg, Asp, Met, Thr, His, Orn, Asn, Cys, Gin, GlyA or LacA. 4 . The sweetener according to claim 2 or 3 , wherein the compound i ,'i :u.'Iect:o(i from y-Glu-Val-Gl y and y-Gl \i-Abu-Gl y .

5. The sweetener according to any one of claimH 1 to 4, wherein the sweet substance comprises one or more kinds of substance selected from the group consisting of aspartame, sucralose, acesulfame K, neotame, N-[N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-a-aspartyl]-L-phenylalanine 1-raethyl ester, saccharin, stevia, glycyrrhizin, thaumatin and monellin. 6 . Anadditiveagenttobeaddedtoasweetsubstance for improving a body taste of the sweet substance, which comprises a compound having calcium receptor-activating effect. 7.An additive agent to be added to a sweet substance for improving a bitter taste of the sweet substance, which comprises a compound having calcium receptor-activating effect.

DESCRIPTION SWEETENER
Technical Field
[0001] The present invention relates to a sweetener and an additive agent for improving the body taste of a sweet substance used as an ingredient of the sweetener, and improving the bitter taste typical of the sweet substance.
Background Art
[0002] In recent years, a high intensity sweetener has been
utilized as a sweetener in a wide range of food field such as diet
foods because of its high sweetness magnification. In the fields
of beverages and confectioneries each consuming an especially large
amount of sweeteners, various high intensity sweeteners have been
used as so-called lower-calorie sweeteners such as a calorie-reduced
sweetener and a calorie-free sweetener, and sugar-free sweeteners.
Of those, typical examples of the high intensity aweetener to be
used include aspartame (APM), sucralose, acesulfame-K (Ace-K),
neotame,
N-[N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-a-aspartyl]-L-phenyl
alanine 1-methyl ester (hereinafter abbreviated as "ANS9801"),
saccharin, stevia, glycyrrhizin, thaumatin and monellin.
[0003]
However, in general, consumers have bocomo f^iml 1 laj' ovor the ynara with the? sweet taste of sugar, glucose ffuatoiin fiyrup or the liko, which haa been used for foods in the pa«l;, (iml bunco ilo not, [Jiolitir in many cases the quality of sweet tasto of: aiich « h I cjh intensity nwMoI.ener as described above. For example, APM hnn a utilization uffect such as a flavor-enhancing effect and an efI.'eat for reducing the bitter taste or the astringency of coexisting Hubstances, while has a problem in its strong after sweet taste (alroncj swoot taste

in tiCt-.ertaate) and storage stability such as heal; reHlstance. Sucraiose has a high stability, while has a probJoin in its flavor suppression effect and heavy sweet taste in aftertaste, for example. Ace-K has a high stability, while has a heavy taste in middle taste to after sweet taste (heavy sweet taste over aftertaste) and particularly has a strong bitter taste and aatringency, therefore, Ace-K is often used in a combination with other high Int-onsity sweeteners. 10004]
Until now, the use of those high intensity sweeteners in combination is known as a technology for modifying the quality of sweetness of the above-mentioned high intensity sweetener (Patent Document 1). Further, the use of those high intensity sweeteners in a combination wit hot her sweeteners such as trehaloseanderythritol (Patent Document 2) or an a-glucosylated stevia extract (Patent Document 3) and the use in a combination with ei diatary fiber (Patent Document 4) have been reported. However, at the preaunt time, tliese have not yet sufficiently satisfied preferences of consumers who have become familiar with the quality of sweetness of sugar or glucose fructose syrup. [0005]
Meanwhile, the calcium receptor which is also called Calcium Sensing Receptor (CaSR) is a receptor consisting of 1,078 amino acids, which is classified into the class C of seven-transmembrane receptors (G protein-coupled receptor; GPCR). Cloning of the gene for the calcium receptor was reported in 1993 (Non-patent document 1) , and the calcium receptor is known to cause various cell responses via elevation of intracellular calcium level etc. , when being activated with calcium etc. The nucleotide sequence of the human calcium receptor is registered with GenBank Accession No. NM 000388 and is well conserved in animals. [0006]
The above-mentioned calcium receptor may act to promote or suppress biological functions . Therefore, at present, a therapeutic

agent that utilizes an activating effect on the calcium receptor nnd a therapeutic agent that utilizes an inh i I) i t(»iy crfoct on the en lei inn receptor are appropriately used in t:,h«' I i iviliii, t«.JO|Micl i vely. Il'al.ont Document .1] JP 2005-304440 A [I'fU.unt Document 2] JP 2002-51723 A (I'al.cnL Document 3] JP 2002-34 501 A |l'nl:i>nl- Document 4] JP 2004-41118 A [IMtonl. Document 5] US 6,548,09ft
(Patent Document 6] International Publication NO. W039979 pamphlet (Non-patent Document 1] Nature, 1993, Vol. 366(6455), pp. 575-580 [Non-patent Document 2 ] J. Endocrinol. , 2 000, Vol . 165(2) , pp. 17 3-177 [Non-patent Document 3] Eur. J. Pharmacol., 2002, Vol. 447(2-3), pp. 271-278
[Non-patent Document 4 ] Cell Calcium, 2004, Vol. 35(3), pp. 209-216 [Non-patent Document 5] J. Biol. Chem., 2006, Vol. 281(13), pp. 8864-8870
Disclosure of the Invention
Problems to be solved by the Invention
[0010]
Objects of the present invention are: 1) to provide a sweetener in which the sweet taste quality, in particular, the body taste and

bitter taste has improved, especially, a high intensity sweetener in which the sweet taste quality, in particular, the body taste and bitter taste has improved; and 2) to provide an additive agent for improving the body taste of a sweet substance which is used as an ingredient of the Hweetenoi', and improvinci (lui hittei 1 ai.rl.c> typical of the sweet substance.
Means for solving the Problems [0011]
The inventors of the present invention found various amino acids and peptides as compounds each having a calcium receptor-activating effect in the process of searching a calcium receptor activator, and also found that the amino acids and peptides can improve the quality of sweetness of a high intensity sweetener, thereby accomplished the present invention.
That is, the present invention is as follows:
(1) a sweetener comprising a sweet substance and a compound having calcium receptor-activating effect;
(2) the sweetener according to ( 1) wherein the compound comprises one or more kinds of amino acid or peptide selected from the group consisting of y-Glu-X-Gly where X represents an amino acid or an dinliio acid derivati.ve, y-Glu-Val-Y where Y represents an amino acid or an amino acid derivative, Y~Glu-Ala, Y~Glu-G],y, Y~Glu-Cys, y-Glu-Met, y-Glu-Thr, y-Glu-Val, Y-Glu-Orn, Asp-Gly, Cys-Gly, Cys-Met, Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, y-Glu-Met(0), Y-Glu-Y-Glu-Val, y-Glu-Val-NHz, Y-Glu-Val-ol, y-Glu-Ser, y-Glu-Tau, Y-Glu-Cys(S-Me) (0) , Y-Glu-Leu, Y-G1U-I.1 c, y-Glii-t-T.eu and
Y-GIn-CyB(S-Me) ;
(:i) the sweetener according to (2) wherein X rt![)i (iH(iHil:H (,'yB(SNO),
CyH (S-a I 1 yX) , Gly, Cys(S-Me), Abu, t-Leu, CIH, All), Pon oi' Ser, and
Y I (.iproaenl.a Gly, Val , Glu, Lys, Phe, Ser, l'it», Aiij, Asp, MeL, Thr,
n I ti, Orn, Ann, Cya, Gin, GlyA or LacA;
(4) the Hwoetenat: according to (2) or (3) whore hi I.ha amino acid
or peptide is selected from y-Glu-Val-Gly and Y-^'Ju-Abu-Gly;

(')) l.he HweeLeiioi; according to any ono ot (1) to (4) whoi.t)Jn the Hweet substance comprises one or more kinds of substance selected from the group consisting of aspartame, sucralotu:), acesulfame K, neotame, ANS9801, saccharin, stevia, glycyrrhizin, thaumatin, monatin and monellin;
(6) an additive agent comprising a compound having calcium receptor-activating effect, which is added to a sweet substance for improving a body taste of the sweet substance; and
(7) an additive agent comprising a compound having calcium receptor-activating effect, which is added to a sweet substance for improving a bitter taste of the sweet substance.
Brief Description of the Drawings [0012]
FIG. 1 is a graph showing an effect of calcium on a calcium receptor . cRNAof the human calcium receptor was injected into oocyte of Xenopus laevis by microinjection. Valuos of intracellular response currents which flew at the time of addition ot a calcium chloride solution at an arbitrary concentration were recorded. The maximum values of intracellular currents were considered response current values. It was confirmed that no response was observed in oocytes in jec ted with distilled water by microinjection as a control.
FIG. 2 is a graph showing an effect of an L-amino acid on a calcium receptor. cRNA of the human calcium receptor was injected into oocyte of Xenopus laevis by microinjection. Values of intracellular response currents which flew at the time of addition of a 10 mM L-amino acid solution were recorded. The maximum values of intracellular currents were considered response current values. It was confirmed that no response was observed in oocytes injected with distilled water by microinjection as a control.
FIG. 3 is a graph showing an effect of a D-amino acid on a calcium receptor. cRNA of the human calcium receptor was injected into oocyte of Xenopus laevis by microinjection. Values of intracellular response currents which flew at the time of addition

of a 10 mM D-amino acid solution were recorded. The maximum values of intracellular currents were considered response current values. 11 wHM confirmed that no response was observed in oocytoH injected Willi (liHiilL(Hl water by microinjection an ^i coiiHol.
FIG. A in a graph showing an effect of n iio|il liin on n c:nlcium I ocept.oi:. cl-, and l.lin abscissa axis repre-sents Iho tlmci jift.or tho start oJ: eating. However, FIG. 5 is a conceptual diagram and does not HIVIW absolvile values. (OOlfiJ
The term "calcium receptor" as used hercjln rofruH to a receptor l.liat ia cai.lfid Calcium Sensing Receptor (CaLiR) and belonga to the class Cof seven-transmembrane receptors . The term "calciumreceptor activator" refers to a substance that binds to tho above-mentioned calcium receptor to activate the calcium receptor. Further, the phrase "to activate a calcium receptor" as used herein means that a ligand binds to a calcium receptor to activate a guanine nucleotide binding protein, thereby transmit a signal. Further, the term "calcium receptor activity" means that the calcium receptor transmits the signal.

Herein, each amino acid and amino acids forming each peptide are L-amino acids unless otherwise stated. [0017] <1> Compound having a calcium receptor-activating effect
The compound having a calcium receptor-activat.ing effect can be an amino acid, a peptide or derivativ^H Ihoroof, ov various low molecular weight compounds as long as the compounti has an effect of improving the sweet taste quality of the sweet substance. The compound can also be a novel compound obtained by screening. For example, the compound can be obtained by reacting a calcium receptor with a test substance, and detecting a calcium receptor activity. It is preferable to confirm that the obtained compound has an effect of improving the quality of sweetness of the sweet substance, in particular, the high intensity sweetener. [0018]
Hereinafter, amethodof screening thecompound having acalciura receptor-activating effect is specifically described, but is not limited to the following steps:
1) measuring a calcium receptor activity by adding a test substance
to a calcium receptor activity measurement system for measuring the
calcium receptor activity;
2) comparing the calcium receptor activity when adding the test substance with a calcium receptor activity when not adding the test substance; and
3) selecting the test substance exhibiting a calcium receptor-activating effect at the time of addition of the test substance.
[0019]
The calcium receptor activity is meaHiuod l)y a monsurement Hystom using cells that express calcium recoptorH, Itir oxamplo. The abovo-montioned cells can be cells endogenou»ly oxpliuin i nq calcium I oiMipl.oi: H, <3i' can bo recombinant cells in wh iiih «n «)X(HJOIU»UH calcium I tHUiptoi' gene is introduced. The calcium lacHptui activity imuiHuremenl:. ays torn described above may be used wi thoul nny particular
10

I i III i lat ion a.s Jong as, when an extracellular' 11(:|MIUI (itct i vnt oi') H|)i!ci |- ic to a calcium receptor is added to t.h(< ahovn' innnl i oncni col 1H that express calcium receptors, the measurement Hyutem may detect the binding (reaction ) between the activator and theca l.cium receptor, or may respond to the binding (reaction) betwecni tho activator and the calcium receptor to thereby transmit a detectable signal into the cells. When the calcium receptor activity is detected through the reaction with the test substance, it is clarified that the test substance has a calcium receptor-stimulating activity, and is a substance that may improve the sweet taste quality of the sweet substance. [0020]
Meanwhile, an effect of improving the quality of sweetness of the sweet substance can be confirmed by a method such as a taste test by humans. Further, the amino acid and peptide to be used as a test substance are not particularly limited; however, the peptide is preferably a peptide consisting of 2 to 10 amino acid residues, or a derivative thereof, and more preferably a peptide consisting of 2 or 3 amino acid roHidues, or a dei ivat ivo thuioul. Furthtir , the amino acid residue at the N-terminal of the peptide is preferably y-glutamic acid. [0021]
Examples of the calcium receptor mentioned above include a calcium receptor derive from an animal such as a mouse, rat and dog as well as human, and the origin of the calcium receptor is not particularly limited. [0022]
As described above, the calcium receptor activity can be confirmed by using live cells expressing a calcium receptor or its fragment, cell membranes expressing a calcium receptor or its fragment, an in vitro system containing a calcium receptor or its fragment protein, or the like.
An example using live cells is described below. However, confirmation of the calcium receptor activity is not limited to this
11

example. [0023]
A calcium receptor is expressed in cultured cells such as those of Xonopus lanvis oocytes, hamster ovarian cells and human fetal kidnoy ce 1.1 s. The calcium receptor can be H>cpf«HtM.iti by cloning a t:iil(.;iiiin iocc!pt:or gene in a plasmid that caifimi « (nioii.in qiMie and i nl i.oduc i nq the plasmid or cRNA obtained by \iHiiii| Uw pl(»Hmid as n liimplat.e l.o tlm eel la. 'J'o detect the iiwurt, l(.»n, «n <» huM I (jfihyiii oloqica 1, technique and a Fliioi i»HCf>nt indiiintoi that 111(11 (uitr-.s nn IncroaHB in intracoll ular calcium lovi'l an amino acid or a peptide having an effect of improving the body taste and/or bitterness of the sweet substance, when the amino acid or the peptide is eaten
12

with the sweet substance. Examples of such an amino acid or a peptide include y-Glu-X-Gly where X represents an amino acid or an amino acid derivative, y-Glu-Val-Y where Y represents an amino acid or anamiiioacidderivative, Y-Glu-Ala, y-Glu-Gly, y-Glu-Cys, y-Glu-Met, Y-Glu-Thr,y-Glu-VaI,y-Glu-Orn, Asp-Gly,Cys-Gly,Cys-Met,Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, y-Glu-Met(0), y-Glu-y-Glu-Val, Y-Glu-Val-NH2, y-Glu-Val-ol, y-Glu-Ser, y-Glu-Tau, Y-Glu-Cys(S-Me) (0), y-Glu-Leu, y-Glu-Ile, y-Glu-t-Leu and y-Glu-Cys(S-Me) (hereinafter, also referred to as a "peptide or the ] ike of the present invention" ) . In the present invention, one kind of those peptides or the like can be used, or two or more k:i nda thereof can be used in combination. [0026]
Further, the peptide may be a peptide derivative having a structure of y-Glu-X-OCH(Z) CO^H (X represents an amino acid or an amino acid derivative, and Z represents H (a hydrogen atom) or CHi (a methyl group) ) . Further, the peptide may bo a compound in which
Y leiJroHenta GlyA or LacA in the formula y-Gl U-VM I -V. I'l n (frable
ajieci ric examples thereof include y-Glu-Va 1-'MyA, \, -G I u--t:l.cni-G l.yA,
Y --(M ii-Abii-( 11 yA, y-Glu-Val-LacA, Y-Glu-tLeU"l,.w:A .nid -, -di I u-Abu-LacA .
II Hhould bo noted that GlyA repreaenta q I yco I i c «ci^i.l, MIHI l.acA
niiMOHonlH b^j'^iywibw acid. B+Hry-i?Tt€ acid may \m orin n[ K-b»^'t•yr^•-r acid
m l<-bntYf-ir'J aciil, and preferrfcl is S-hiH-y r io /inid. SI rnot u la I
formulae of those compounds are described below.
I 0021 I


0v^^^°2"
HOj,C

0028]

13

H3C,

CH,

:cH,

HOjC
NH,

Ov. ^COjH

[0029]


0.. ^COjH
HOjC

[0030]


H3C.
HOjC
CH,

0031]

HOjC

0 CH,
Os,^^COjH

0032]

14


Os^^COjH
HOj,C

3
GH

[0033]
Herein, examples of the amino acid includo a neutral amino acid such as Gly, Ala, Val, Leu, lie, Ser, Thr, Cys, Met, Aan, Gin, Pro, Hyp and t-Leu, an acidic amino acid such as Aap ond Glu, a basic amino acid such as Lys, Arg and His, an aromatic amino acid such as Phe, Tyr and Trp, and homoserine, citrulline, ornithine, a-aminobutyric acid, norvaline, norleucine and taurine. The amino acid can also be a non-naturally occurring ( non-protein constituent) amino acid such as tert-leucine, cycloleucine, a-aminoisobutyric acid and L-penicillamine. It should be noted that X in the peptide Y-Glu-X-Gly canbe any one of above-mentioned amino acid or a derivative thereof, and preferably an amino acid or a derivative tlxoreof, other than Cys, [0034]
Herein,abbreviationsforaminoacidresiduesmean the following amino acids.
(1) Gly: Glycine
(2) Ala: Alanine
(3) Val: Valine
(4) Leu: Leucine
(5) lie: Isoleucine
(6) Met: Methionine
(7) Phe: Phenylalanine
(8) Tyr: Tyrosine
(9) Trp: Tryptophan

(10) His: Histidine
(11) Lys: Lysine
15

(12)
(13)
(14)
(15)
( 1 6 )
(17)
(IB)
( 1'))
(■/O)
(2 1 )
( .' 2 )
(2J)
(24)
(2b)
(:■' ci)
(27)
(28)
(29)
(30)
(31)
(32)
|003
Arg: Arginine
Ser: Serine
Thr: Threonine
Asp: Aspartic acid
Glu: Glutamic acid
Ann: Asparagine
iMn: (Vlul.amine
Cya: CynLeine
['I'o ! I Mo I ino
()i.n! Ornithine
i\nv : 'n»rCOB i no
Cit: Citrulline
N-Vcil : Norvaline
N-Leu: Norleucine
Abu: a-Aminobutyric acid
'I'au: Taui-ine
Hyp: Hydroxyproline
t-Leu: tert-Leucine
Cle: Cycloleucine
Aib: a-Aminoisobutyric acid {2-methylalanine)
Pen: L-Penicillamine
yFurther, examples of the amino acid deri vnti ve i nc 1 ude various derivatives of above-mentioned amino acids such aw a special amino acid, a non-natural amino acid, an amino alcohol and a substituted amino acid of which an amino acid side chain such as the terminal carbonyl group, the terminal amino group, and the thiol group of cysteine, is substituted with various substituents. Examples of the substituent include an alkyl group, an acyl group, a hydroxy group, an amino group, an alkylamino group, a nitro group, a sulfonyl group and various protective groups, specifically include Arg(NO2) : N-Y-nitroarginine, Cy8(SN0): S-nitrocysteine, Cya(S-Me): S-methylcysteine, Cys ( S-allyl) : S-ally.l cyHtulne, Vnl-NH;;! valinamide, Val-ol: vaiinol (2-amino-3-methyl-l-butanol).
16

[0036]
It should be noted that herein, y-Glu-Cys(SNO)-Gly has the following structural formula, and the "(0)" in the above formulae Y-Glu-Met(O) andY-Glu-Cys(S-Me) (0) indicatesasulf oxide structure. The "(Y)" in the formula y-Glu indicates that glutamic acid binds to another amino acid via the carboxy group at the y-position in glutamic acid. [0037]
COjH
IIO,C
:0
NMj 0
S-Nltro80glutath(one (QNSO)
I 0()3fl ]
Y~Glu-X-Gly where X represents an amino acid ot an nmino acid 'li'i Ivrii i vc, Y-Glu-Val-Y where Y representH an flminn nc.id or nn amino ,11 I (I (li;r 1 vdl i vo, Y-Gl-U-AUi, Y~Glu-Gly, y-Glu-Cyii, Y"'^»l"-Mot, y - III u-Tlir, Y-^:']"~V.il r v-Glu-Om, Asp-Gly , Cya Gl y , Cyn Mrd , G I u-Cy.s , i;)ly-Cyu, Iieu-Asp, D-Cys, Y-Gl>J-MQt(0) , Y-^jhi-y-GI u-Vfl 1 , Y-Glu-Val-NH2, Y-Glu-Val-ol, y-Glu-Ser, y-Glu-Tau, Y-Glu-Cy8 (S-Me) (0) , y-Glu-Leu, Y~G1"~I1*>/ Y-Glu--I.-I.eu and y-GlU"Cyn ( S-MG) each improve the body haflte and 1)1 l.ternesB of the Hweet substance.
Therefore, yGlu-X-Gly where X representH an amino acid or an amino acid derivative, y-Glu-Val-Y where Y represents an amino acid or an amino acid derivative, y-Glu-Ala, y-Gl-ii-^ly» y-Glu-Cys, Y-Glu-Met, Y-Glu-Thr, y-Glu-Val, y-Glu-Orn, Aap-Gly, Cys-Gly, Cya-Met, Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, y-Glu-Met(0), Y-Glu-Y-Glu-Val, y-Glu-Val-NHz, y-Glu-Val-ol, y-GJ-ii-Ser, y-Glu-Tau,
17

Y-Glu-Cys(S-Me)(0), y-Glu-Leu, y-Glu-Ile, y-GIu-t-Leu and y-Glu-Cys ( S-Me) (hereinafter, also referred to as "peptide and amino acid to be used in the present invention" ) can be used as an additive agent for a sweet substance which is added to a sweet substance for improving the body taste and bitterness of the sweet substance. [0039]
The compound to be used in the present invention can be used alone or as a mixture of arbitrary two oj more kindn. Of those, a compound having the following struct; ur a 1 Im iiui 1 -i; y ■'• I »i -K-clly wheio X represents Cys(SNO), Cys(S-allyl), Gly, CyB(S-Me), Abu, t-Leu, Cle, Aib, Pen or Ser, or y-Glu-Val-Y where Y represents Gly, Val, Glu, Lys, Phe, Ser, Pro, Arg, Asp, Met, Thr, His, Orn, Asn, Cys, Gin, GlyA or LacA is preferably used.
Of those compounds, preferred compounds are y-Glu-Val-Gly and Y-Glu-Abu-Gly. [0040]
As the above-mentioned compound to be used in the present invention, a compound which is commercially-available can be used. Further, when the compound was a peptide, the peptide can be obtained by appropriately using a known technique such as (1) a method of chemically synthesizing the peptide or (2) a method of synthesizing the peptide by an enzymatic reaction. Since the number of amino acid residues contained in the peptide to be uacid in hhe present invention is as comparatively small as 2 or 3 residues, a method of chemically synthesizing the peptide is convenient. When the peptideischemicallysynthesized, theoligopeptidecanbe synthesized or semi-synthesized by using a peptide synthesizer. An example of the method of chemically synthesizing the peptide includes a peptide solid phase synthetic method. The peptide synthesized as described abovecanbepurified by usual means such as ion exchange chromatography, reversed phase high performance liquid chromatogiraphy or affinity chromatography. Such a peptide solid phoHH Bynlh«»l;lo method and I.he subsequent peptide purification are well known in l.luj technical ( 1 >3 \ cl .

I 0 0 4 1 I
Purthor, Lhe pefitide to be used in i:ho |vrefloiil i iivnrit iun can also be produced by an enzymatic reaction. For exampl.e, the method (l(>Hcribed in International Publication NO. WO2004/0 I I 6!:),l pamphlet (.^ati be UBod. 'I'hat is, the peptide can also bo prudiicMuJ by reacting an amino acid or dipeptide in which carboxyl termlnuH is esterified or amidated with an amino acid having a free amino group { for example, an amino acid in which carboxyl group is protected) In the presence of a peptide producing enzyme, and purifying the produced dipeptide or tripeptide. Examples of the peptide producing enzyme include a culture of a microorganism having an ability to produce a peptide, microbial cells separated from the culture or a procoBsed product of cells of the microorganism, or a peptide produciny un/yme derived from the microorganism.
It should be noted that the peptide to be used in the present invention is not only produced by such an enzymatic method or a chemical synthesismethodasmentionedabove, butalsomayexistin, forexample, a plant such as a vegetable or a fruit, a microorganism such as a yeast andayeast extract. Whenthepeptideexists in naturalproducts, the peptide which is extracted from those natural products can be used.
Further, the peptide does not need tu 1)0 iHolat-od before use, and there can be used as a fraction containing the peptide of the present invention in a large amount. [0042]
Examples of the low molecular weight compound to be used in the present invention include cinacalcet
((R)-N-(3-(3-(trifluoromethyl)phenyl)propyl)-!-{1-naphthyl)ethy lamine) and an analogous compound thereof . Examples of the analogous compound of cinacalcet include the compound represented by the following chemical formula (1);
((R)-N-[(4-ethoxy-3-methylphenyl)methyl]-l-(1-naphthyl)ethylami ne) ), or the compound represented by the following chemical formula (2);
19

( ( R)-N- ( 3-phenylprop-2-enyl) -l-( 3-methoxyphenyl )eLhy 1.amine ) . Those compounds can be synthesized by such a known method as described in US Patent No. 6,211,244, for example. Further, commercially-available products can also be used. [0043]

^==^'

(1)

r-'X
,^^^
(2)
10044]
The compound to be used in the present invention also includes that in the form of a salt. When the peptide and the amino acid to be used in the present invention are in the form of a salt, the salt can be a pharmacologically acceptable salt. Blxamples of a salt with an acidic group such as a carboxyl group in the formula include an ammonium salt, a salt with an alkali metal such as sodium and potassium, a salt with an alkaline earth metal such as calcium and magnesium, an aluminum salt, a zinc salt, a Ba.lt with an organic amine such as triethylamine, ethanolamine, morpholine, pyrrolidine, piperidine, piperazine and dicyclohexylamine, and a salt with a basic amino acid such as arginine and lysine. Examples of a salt with a basic group in case where the basic group exists in the formula include a salt with an inorganic acid such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid and hydrobromic acid, a salt with an organic carboxylic acid such as acetic acid, citric acid, benzoic acid, maleic acid, f umaric ac lil, Inrtarlaflcld, succinic
20

acid, tannic acid, butyric acid, hibenzoic acid, pamoic acid,
enanthoic acid, decanoic acid, teoclic acid, salicylic acid, lactic
acid, oxalic acid, mandelic acid and malic acid, and a salt with
anorganicsulfonicacidsuchas methanesulfonicacid,benzenesulfonic
acid and p-toluenesulfonic acid.
[0045]
<2> Additive agent and sweetener of present invention
The compound to be used in the present invention, preferably the peptide and the amino acid, can be used as an additive agent which is added to a sweet substance in order to improve the body taste of the sweet substance. The peptide and the amino acid to be used in the present invention can also be used ao an additive agent which is added to a sweet substance in order to improve the bitterness of the sweet substance.
The compound can be used alone or as a mixture of two or more kinds. The compound to be used in the present invention can also be mixed with a sweet substance to form a sweetener. [0046]
The additive agent of the present invention can consist of only one kind or two or more kinds of compovnuliii crhoHon from the ttbove-mon t i on ed compound to be used in theproH»nt; 1 nvi»nl: jon . Further, t.ho additive agent can be formed by optional 1 y fldd I ng ol lun compounds 1)1 vui iuun addii. iveH each liavlng an effect of I mpinv i in| I tu» qunlity ol HwnelnoHH of tlie sweetener. I 004 7]
The amount of the additive agent which is added to the sweet HubtiLcince LB not particularly limited as Tnng«H thenmount can improve the quality of sweetness of the sweet substance, in particular, the l)ody taste . Specific amount of the compound of the putMont 1 nvontion IH, for example, 0.000001% to 99 . 9999% by weight, pref ornbly 0 . 0000 1% to 99.999% by weight, and more preferably 0.0001% to 99.99% by weight with respect to the high intensity sweetener.
It should be noted that the above description is a description about the mixture proportion with respect to the high intensity
21

sweetener, and the use amount of the compound of tho prnHont invent ion in a food or a beverage is 1 ppb to 99.9% by weigiil, preferabJy 1.0 ppb to 10% by weight, and more preferably 1 ppm to 1% by weight. [0048]
The sweetener of the present invention contains the sweet substance and the compound to be used in the present invention. The compound can be used alone or as a mixture of two or more kinds.
As the sweet substance, a high intensity sweetener is preferably used, and examples thereof include aspartame, sucraioHe, acesulfame K, neotame, ANS9801, saccharin, stevia, ql ycyrrli.l z 1 n, thaumatin, monatin andmonellin. Of those, aspartame, aucraioHi! and acesulfame K are preferably used.
The sweet substance can be used alone or as a mi.xture of two or more kinds . It is known that themixture of mult iple.sweet substances can improve the body taste of each of the sweet substances. Even in such a case, the combined use of the peptide or the amino acid to be used in the present invention with the sweet substances can further improve the body taste. Examples of the preferred combination of the sweet substances include a combination of two kinds optionally selected from aspartame, sucralose andacesulf ame-K, or a combination of those three kinds.
Further, an example of the preferred combination includes a combination of aspartame and y-Glu-Val-Gly which is considered to have a high effect in the present invention. Specifically, these can be mixed so that the weight proportion of y-'^l^-'-Val-Gly might be 0.1 to 500 parts by weight with respect to 100 parts by weight of aspartame. As a matter of course, this is merely one example of mixture examples, and there is no need to be restricted to the above-mentioned proportion.
The sweetener of the present invention can of course be mixed with common sweeteners such as sucrose, glucose, fructose and sugar alcohols (for example, erythritol and maltitol). Examples of the form of the sweetener of the present invention Innludo a powder, a granule and a liquid, and the physical propertIHH nvu not limited.
22

I'lxamp Lew 10 0-19 I
Hereinafter, the present invention IH moi e Hpt^c i Lica lJ.y described with reference to examples. Howovor, thn Hoopo of the IJiftHGiil: invention is not limited to thoa«! oxamtileH. I, OObO] [Reference Example 1] Preparation of calcium receptor gene (cRNA)
The gene of the calcium receptor was prepared as follows. On the basis of the DNA sequence registered at NCBl (calciuin receptors NM 000388), synthetic oligo DNAs (forward primer (SEQ ID NOj 1) and reverse primer (SEQ ID NO: 2)) used for PCR were synthesized. [0051]
PCR was performed by using the primers and I'fu ultra DNA Polymerase (manufactured by Stratagene) under tlui following conditions by using human kidney cDNA (manufactured by Clontech) as a material. After a reaction at 94°C for 3 minutes, a cycle of reactions at 94°C for 30 seconds, 55°C for 30 seconds and 72°C for 2 minutes was repeated 35 times, and then a reaction was performed at 72°C for 7 minutes. Whether amplification was attained by PCR was confirmed by performing agarose electrophoresis of DNA, staining the DNA with a DNA staining reagent, and subsequent ultraviolet irradialion. The chain Jongths of t:hr.' VCR |irnductH wnro confirmed by comparison with DNAmarkers of known sizes simultaneouBly subjected to the electrophoresis . The plasmid vector pBR322 was digested with the restriction enzyme EcoRV (manufactured by Takara). The gene fragment amplified by PCR was ligated to the cleavage site of the plasmid by using Ligation Kit (manufactured by Promega). The Escherichia coli DHScx strain was transformed with each ligation reaction solution, and a transformant harboring the plasmid in which the PCR amplification product was cloned was selected. The PCR amplification product was confirmed by DNA base sequence analysis. By using the recombinant plasmid as a template together with a cRNA preparation kit (manuf acturedby Ambion) , cRNAof thecalciumreceptor
23

gene was prepared.
[00 52 I
I Reference Example 2] Preparation of various samples
As L-amino acid sample, 23 kinds of special grade amino acids including alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, l.ryptophan, tyrosine, valine, ornithine and taurine (all of those Irom A j Lnomotc) Co ., Tnc . ) , and hydroxypro I inn { Nvic:a:i: n I 'I'ISHCIUO , Tnc . ) , wcne used. AH D-Cya and D-Trp (Nacarai Toni|U(i, lim.) /\H(I calcium I'll 1 111" id(>, tluiHe of special grade were uHod, I 0 0') J j
l''uithnr , aH p«,)ptide sample, y-Glu-Cyn-Gl y ( H I qmn h 11\\ icli Japan X . K . ) , Y-('■ 1 u-(..;y8 ( SNO) Gly ( Do j indo Laborator 1 OH ) , \ ■ tl 1 u-A1 n (llachem l''o.Liichemi kaJ, ien AG), y^lu-Gly (Bachem Feinchem Ikalien AG), y-C'ilu-CyH (Sigma Aldrich Japan K.K.), y-Glu-Mot (Unchem FeinchGiTii kdl i.enA(j) , y-Glu-Abu-Gly (Abu: a-amlnobuiyrlcacid, Bachem I'l'l iK^hern i. kfi.l i en AG), y-Glu-Thr (Koltusan Ghomica I Co., litd . ) , y-Glu-Vai ( Kokusan Chemical Co . , Ltd.), y-Glu-Leu (cnuttom nynthesis product), Y-Glu-Ile (custom synthesis product) , y-Glu-Orn (Kokusan Chemical Co., Ltd.), Asp-Gly (custom synthesis product), Cys-Gly (custom synthesis product), Cys-Met (custom syntliosis product), Glu-Cys (custom synthesis product), Gly-Cys (cuHl.om fjynthesis product), Leu-Asp (custom synthesis product) , Y-Gl-U~Val-Val (custom synthesis product), y-Glu-Val-Glu (custom syntheHia product), y-Glu-Val-Lys (custom synthesis product) , y-Glu-y-Glu-Val (custom synthesis product), y-Glu-Gly-Gly (custom syntheHia product), Y-Glu-Val-Phe (custom synthesis product), Y~Glu-Val-Ser (custom synthesis product), y-Glu-Val-Pro (custom synthesis product), Y-Glu-Val-Arg (custom synthesis product), y-Glu-Val-Asp (custom synthesis product), y-Glu-Val-Met (custom synthesis product), Y-Glu-Val-Thr (custom synthesis product), y-Gl^-Val-His (custom synthesis product), y-Glu-Val-Asn (custom synthesis product), Y-Glu-Val-GIn (custom synthesis product), y-Glu-Val-Cys (custom
24

synthesis product), y-Glu-Val-Orn (custom synthesis product) and y-Glu-Ser-Gly (custom synthesis product) were used. Glutamine and cysteine were prepared upon use, and the other samples were stored at -20°C after preparation. The peptides having a purity of 90% or higher were used. As only for y-Glu-Cys, one having a purity of 80% or higher was used. [0054]
After dissolving each sample in solution, pH of the solution showing an acidic or alkaline pH was adjusted to an approximately neutral pH by using NaOH or HCl. The composition of the solution used for dissolution of amino acids and peptides, and the solution used for preparation of Xenopus laevis oocytes and culture of the oocytes is as follows. 96 mM NaCl, 2 mM KCl, 1 mM MgCl^., 1,8 mM CaClz, 5 mM Hepes, and pH 7.2. I 0055] [Reference Example 3] Synthesis of y-Glu-Val-Gly
Boc-Val~OH (8.69 g, 40.0 mmol) and Gly-OBzl'HCl (8.07 g, 40.0 mmol) were dissolved in methylene chloride (100 ml) , and the solution was kept at 0"C. Triethylamine (6.13 ml, 44.0 mmol), HOBt (1-hydroxybenzotriazole, 6.74 g, 44.0 mmol) and WSC-HCl (1-ethy1-3-(3-dimethylaminopropyl)carbodlimide hydrochloride, li.4^ g, 44.0 mmol) were added to the solution, and ( hfj mlxi.iue was Hl..i rred ovfMnight ai. room tempera Lure. The rr.an\ Ion no hit ion was ('(iiitinnl.rated under reduced pressure, and Lh« roHldim wan dloHolved in ot.hyl acetate (200 ml). The solution waa waBhrid with water (50 ml ), '>* of cil.rlc acid aqueous solution C^iO ml *■ Iwloo). Hfituratod brine (50 ml) , 5t of sodium hydrogen cnrbonflt.o nquiiiouH BoluLlon (50 ml X twice), and saturated brine (50 ml). The organic layer was dried over anhydrous magnesium sulfate, magnauliimiuu 1 f nt.o was removed by filtration, and the filtrate was concentrated undtir reduced piesaure. Tlie residue was recrystallized from isl.hyl acetate/n-hexane to obtain Boc-Val-Gly-ODzl (13.2 g, 36.2 mmol) as a white crystal. [0056]
25

Boc-Val-Gly-OBzl (5.47g, IS.Ommol) waaadded l.o 4N11C1 /dioxane solution (40 ml), and the mixture was stirred at room l.amperatiire lor 50 minutes. Dioxane was removed by concentraLi t^n under reduced pressure, n-hexane (30 ml) was added to the residue, and the mixture was concentrated under reduced pressure. The procedure was repeated 3 times to quantitatively obtain H-Val-Gly-OBzl'HCl.
H-Val-Gly-OBzl•HClandZ-Glu-OBzl(5.57 g,15.Ommol)described above was dissolved in methylene chloride (50 ml), and the solution was kept at 0°C. Triethylamine (2.30 ml, 16.5 mmol), HOBt (1-hydroxybenzotriazole, 2.53 g, 16.5 mmol) and WSC-flCl (l-ethyl-3-( 3-dimethylamLnopropyl )carbodi imido hydi.Tichloride, 3.16 g, 16.5 mmol) were added to the soiution, and I lie mixture was stirred at room temperature overnight for 2 days. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in heated ethyl acetate (1,500 ml) . The solution was washed with water (200 ml), 5% of citric acid aqueous solution (200 ml X twice), saturated brine (150 ml), 5% sodium hydrogen carbonate aqueous solution (200 ml x twice) and saturated brine (150 ml). The organic layer was dried over anhydrous magnesium sulfate, magnesium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure. The precipitated crystal was collected by filtration and dried under reduced pressure to obtain Z-Glu(Val-Gly-ODzl)-OBzl (6.51 g, 10,5 mmol) as a white crystal. [0057]
Z-Glu(Val-Gly-0Bzl)-0B2l (6.20 g, 10.03 mmol ) described above was suspended in ethanol (200 ml), 10% palladium on carbon (1.50 g) was added to the suspension, and a reduction reaction was performed under a hydrogen atmosphere at 55 °C for 5 hours . During the reaction , 100 ml in a total volume of water were gradually added. The catalyst was removed by filtration using a Kiriyama funnel, and the filtrate was concentratedunder reducedpressure to a half volume. The reaction solution was further filtered through a membrane filter, and the filtrate was concentrated under reduced pressure. The residue was di.HHolved in a small volume of water, and to t:.ho Holution, ethanol
26

wiin iiildocl to pron ip i tale a crystal, and t.hu (nyHl.nl wnn UD], lee: tod
I'V ' ' I I: I .11 11 riLi;al.i on-dopendeni: CI ionic cuirenl. imuinui i mj inul hod uHing a StinopiiH laavis oocyte expression system wnH UHod, it ncLivator In (UlcUid Lo X(tMo/)i/.'( letevia oocyteH .in whl<^h t ho iicUcium racept.or .is expressed, intracellular Ca ions increase. Then, the Ca ion (Kjncontration-dependent CI channel opens, and the Intracellular current value changes as an ionic current. Uy meaHurijig Iho change -In the intracellular current value, whether the fmlcium fnceptor-actlvating effect is present or not cnu be comfIrmed. [0063]
Specifically, abdomen of Xenopus laevia was opened, and an eggbaLcliwas taken out and then treated with a 1 % co I I agnnaBe solution at 20"C for 2 hours to obtain individual oocytes. Into each oocyte, 50 n]. of 1 |jg/)jl receptor cRNA or 50 nl of sterilized water per oocyte were injected by using a micro glass capillary, and the oocytes were cultured at 18 "C for 2 to 3 days. For the culture, a solution obtained by adding 2 mM of pyruvic acid, 10 U/ml of penJoi 1 J in and 10 ijg/nil of streptomycin to the solution mentioned in Reference Example 2 was used. After the culture, a test solution was added to the oocytes injected with cRNA or sterilized water. Electrophysiological measurement was performed by using an amplifier Geneclarap 500 (manufactured by Axon) and recording software AxoScope 9.0 (manufactured by Axon). The oocytes were voltage-clamped at -70
28

mV by the double electrode voltage clamp method, and l;h(! i nl.)acell ular
current via the Ca ion concentration-dependent CI ion was measured.
The maximum value of the intracellular current was considered as
the response current value.
[0064]
[Reference Example 7 ] Evaluation of effect of calcium for activating
calcium receptor
The calcium receptor-activating effect was evaluated by using themethoddescribedinReferenceExample 6. That is,oocytes injected with cRNA of the calcium receptor or sterilized water was prepared, and voltage-clamped at -70 mV by the double electrode voltage clamp method. To the voltage-clamped oocytes, calcium was added (2 mM, 5 mM, 10 mM and 20 mM) , and then Ca ion concentration-dependent Cl response current was measured. FIG. 1 shows the jreaults. From the results, it was confirmed that cRNA of the calcium receptor injected into the oocytes was functionally expressed. Further, because the oocytes in jected with water did not respond to even a high concent ration of calcium, itwasconfirmedthat thecalciumreceptorwas not expressed in the oocytes themselves. [0065]
[Reference Example 8] Evaluation of effect of L-amino acids for activating calcium receptor
'I'ho OIMDCL of L-amino acids for acl.ivnl lnr| cnlciuin teni'ptor Vina ovaliiaLod by using the method describtid In HofHionco Example (i. Thai, id, oocytes injected with cRNA oT t IHI Cfilcluiit iHr^rsptor or (il «(■ i I Lz(:id water ware prepared, and voll.ay«~c I niii|>Md at. -70 mV by I h(! double olactrodc voltage clamp method. To t h»i vo 11 atjo-o lamped oooyl.oH, alanine (10 mM), arginlne (10 mM) , aaimrnghu) { 1 () mM) , aspartic acid (10 mM) , cysteine (10 mM), glutamine ( 10 mM) , glutamic acid (10 mM), glycine (10 mM)', histidino (10 mM) , JHoleucine (10 mM), leucine (lOmM), lysine (lOmM), methionine (lOmM), phenylalanine ( lOmM) , proline ( 10 mM), serine ( 10 mM), threonine ( lOmM) , tryptophan (lOmM), tyrosine (lOmM), valine (lOmM), ornithine (lOmM), taurine (10 mM) or hydroxyproline (10 mM) was added, and Ca ion
29

concentration-dependent Cl response current was niuaHured. FIG. 2 HhowB the results . From the results , it is demons t rated that cysteine, hi.stidine, phenylalanine, tryptophan and tyrosinonach had a def ini to calcium receptor-activating effect. The activatintj effect of the above-mentioned amino acids was reported in Proc. Nntl. Acad. Sci. USA, Apr. 25, 2000, 97(9): 4814-9. [0066]
[Reference Example 9] Evaluation of effect of D-cysteine for activating calcium receptor
The effect of D-cysteine for activating calcium receptor was evaluated by using the method described in Reference Example 6 . That is, oocytes injected with cRNA of the calcium receptor or sterilized water were prepared, and voltage-cl.ampjHi al -70 mV liy I he double electrode voltage clamp method. To the voltage-clamped oocytes, D-cysteine (10 mM), L-cysteine (10 mM), D-tryptophan (10 mM) or L-tryptophan (10 mM) was added, and Ca ion concentration-dependent Cl response current was measured. FIG. 3 shows the results. From the results, it is demonstrated that D-cysteine had a definite calcium receptor-activating effect. [0067]
[Reference Example 10] Evaluation of effect of peptide for activating calcium receptor
The effect of peptide for activating calcium receptor was evaluated by using the method described in Reference Example 6 . That is , oocytes injected with cRNA of the calcium receptor or sterilized water were prepared, and voltage-clamped at -70 mV by the double electrode voltage clamp method. To the voltage-clamped oocytes, y-Glu-Cys-Gly (50 \xM) , y-Glu-Cys( SNO)-Gly (50 pM) , y-Glu-Ala (50 [JM), Y-Glu-Gly (500 pM), y-Glu-Cys (50 pM) , y-Glu-Mel: (500 pM) , Y-Glu-Thr (50 liM) , y-Glu-Val (50 pM) , y-Glu-Orn (500 pM) , Asp-Gly (1 mM), Cys-Gly (1 mM), Cys-Met (1 mM), Glu-Cys (50 pM), Gly-Cys (500pM) or Leu-Asp (ImM) was added, and Ca ion concentration-dependent Cl response current was measured. FIG. 4 shows the results. From the results, it is demonstrated that the above-mentioned peptide
30

luul (1 clelinil.e ef:fect tor activating c^al.(.-iurn rtu'iiptdi.
I Mi» I'm (!nc;o hlxample 1 1. ] Evaliiation ot eftoct: cif |»o|)tlil<« (Di n<:-l Ivalinq t.'M I c; I iiiii I r.M;(!pl,(ji:
The oftect of. peptide for activating calcium rucuptor was ovaJnated in the same manner as that of Reference Flxamplo 10. I'lach of the peptidea shown in Table 1 was added to voliage-u 1 aiiipud oocytes dt 1,000 |JM, .300 |JM, 100 (JM, 30 ^iM, 10 [\M. ( iiM, I \iM, O.) |.iM and 0. I |,iM, .iiicl i.',:i .ion c:oncenti.:ation-dependent CJ loHponHo curi-ent was measured. The lowest concentration with which current was detected was shown in Table 1 as the activity. From the reaultH, it is revealed that those 32 kinds of peptides each had a calcium reaoptor-activating effect. [0069]
31

Table 1
Number
1
2 ~""
3 Peptide
Y-Giu-MetToT_ "2 f 1.
■"Y-GYu-Val-Var
Y-Glu-Val-Glu Activity
1,000 ]]M
1,000 |.iM 1,000 MM
4
^5 '"""" Y-Glu-Val-Lys y-Glu-Val-Arg 1,000 )iM Jl j^OOO JJM
I7OOO"PM"
6 y-Glu-Val-Asp

7 y-Glu-Val-Met 1,000 |JM
8 y-Glu-Val-Thr 1,000 nM
9 y-Glu-y-Glu-Val 1,000 MM
10 y-Glu-Val-NH2 1,000 pM
11
^3 2"'"'"
"]J
M
Ml
u>
17
Ill ) ') ■/{)
). 1
22 2 3 24 y-Glu-Val-ol 1,000 pM

y-Glu-Ser y-Glu-Tau
y-G.lu-CyB (S-Me?) (^^^^^
Y-Glu-Val~Hia y-Glu-Vai-brn jqo jlM
300 jiM
300 |iM
VOO |iM
lOQ |iM
100 |iM
100 |iM
ioO |iM
100 pM
30 |iM
30 |JM
30 |.iM
30 (iM •
30"~iiM'
"30"|'iH
I'O nM
To'fiM
~10 'I'lM ~ '
3 pM

Y-{M»-I,oii
Y-G1u-Tle
y-Gl u-t-IiGU
Y-Clu-CyafS-allyiy-Gly
Y-Glu-Val-Asn
y-Glu-Giy-Gly
y-Glu-Val-Phe
y-Giu-Val-Ser

25 26
2 7 "
'28
"2 y "' ""
'30 y-Glu-Val-Pro y-Glu-Ser-Gly

Y-Glu-Cys(S-Me)

y-Glu-Val-Cys

Y-Glu-Val-Gln

y-Glu-Abu-Gly

31 y-Glu-Cys(S-Me)-Gly

32 y-Glu-Val-Gly 0.1 iiM
[OOVOj
[Reference Example 12] Kokumi-imparting activity of peptide and amino
acid to be used in present invention
Typical examples are selected from y-Glu-X-Gly whore X represents Cys{SNO), Cys(S-allyl), Gly, Cys(S-Me), Abu or Ser, y-Glu-Val-y where Y represents Gly, Val, Glu, Lys, Phe, Ser, Pro, Arg, Asp, Met, Thr, His, Orn, Asn, Cys or Gin, y-Glu-Ala, y-Glu-Gly, Y-Glu-Cys, y-Glu-Met, y-Glu-Thr, y-Glu-Val, y-Glu-Orn, Asp-Gly,
32

Cys-Gly, Cys-Met, Glu-Cys, Gly-Cys, Leu-ABp, D-Cy.s, y-^^ iLi-Met (0) , Y-Glu-y-GJu-Val, y-Glu-Va L-NH2, y-tJ' "-Vnl ■" I , s (i 111 ■ :H. 1 , y-G.lvi-'I'au , Y-Glu-Cys(S-Me) (0) , y-Glu-Leu, y-Glu-Jle, y-Glu-t-l.uu and Y-Glu-Cys(S-Me), ineachof whichacalciumreceptor-activatingef feet was found, andthepresenceor absence of the kokumi-impart ing activity was examined by a sensory evaluation test. [0071]
The sensory evaluation test was performed as follows. To distilled water containing sodium glutamate (0.05 g/dl), inosinic acid monophosphate (0.05 g/dl) and calcium chloride (1 mM), each of alliin (S-allyl-cysteine sulfoxide: control experiment of kokumi-impartingactivity),y-Glu-Cys-Gly, y-Glu-Cys, y-Glu-Alaand Y-Glu-Val was mixed as a sample in an amount of 0.2 g/dl, and the presence or absence of the kokumi-imparting activity was determined. As for a sample which is acidic when the sample is dissolved, pH of the sample was adjusted with NaOH to pH 6.8 to 7.2 before use. Table 2 shows the results. [0072] Table 2 Kokumi-imparting activity of calcium receptor promoter

Calcium receptor promoter Kokumi-imparting activity
Y-Glu-Cys-Gly +
Y-Glu-Cys +
Y-Glu-Ala +
Y-Glu-Val +
I 007.1]
[MeferencG Example 13] Kokumi-imparting acliv.ity of piipl.ido to be
iiHiiil In Lliu proHunt invention
A pepl. ide in which a calcium recepLoi-AIM, I vat I ny of foci was I (Mind wflH oxamined for intensity of its kokumi-.impnri I ricj activity l)y a quanl.l tatl.ve sensory evaluaLion teal..
The quantitative sensory evaluation test wna performed as fo LlowB . To distil led water containing sod iumglutniiin I.o ( 0 .05g/dl), inosinic acid monophosphate (0.05 g/dl) and sodium chloride (0.5 y/dl), eacli of y-Glu-Cys-Gly (glutathione), y-^'l i-'-/**•'"''« Y-^l-U-Met
33

and Y-Glu-Val was mixed as a sample in an amount ol 0.1 g/dl, and Lhe intensity of kokumi-imparting activity was moaHured. Aa for a sample which is acidic when the sample is disHOlvnd, pH of the sample was adjusted with NaOH to pH 6.8 to 7 .2 before UHe. Fiecauae it is known that glutathione can impart kokumi to a IJood, and thus, glutathione was used as a comparative control. 'I'he tertt won performed with n=3 based on the following sensory evaluation acoreii s control : 0 point and glutathione addition : 3 points. Table 3 shows the results . It should be noted that the term "initial and middle taste" is a collective term of an initial taste and a middle taste. [0074]

3.0
0.5
1.5
Table 3
Sample Control
Y-Glu-Cys-Gly
Y-Glu-Ala
Y-Glu-Met

Concentration (g/dl)
0.1
0.1
0.1

Tnil.ial and niUltl.1 f5_ taste 0 '
Kokuwi-impartinq activ^it_y
Alitoi VflHtu
'o_
3.0
0.2
0.4

Tnuta (iiolM, le
'I'lilukimttM, ijrowth, and
cojiUjuii ty ^'■2_'>"'i5i!££5li.
The of feet LB weak, but thickneBo iti slightly
o^nhanoed. .
ThicknBBH rtnd growth are

Y-Glu-Val

0.1

3.0

1.0

Tliicknaas and growth are enhancod mainly in initial and middlo taste.

[0075]
[Reference Example 14] Kokumi-imparting activity of peptide to be
used in the present invention
A peptide in which a calcium receptor-activating effect was found was examined for intensity of its kokumi-imparting activity by a quantitative sensory evaluation test.
The quantitative sensory evaluation test was performed as follows. To distilledwater containing sodi umglutamate(0.05 g/dl), inosinic acid monophosphate (0.05 g/dl) and sodium chloride (0.5 g/dl), each of Y~Glu-Cys-Gly (glutathione) , Y-Gli^-'-Y''< Y~Glu-Val and Y-Glu-Val-Gly was mixed as a sample in an amount of 0.1 g/dl, or 0.01 g/dl as required, and the intensity of kokumi-imparting activity was measured. As for a sample which is acidic when the
34

ti/impln i !t ci i HHcjJ.vc'd, pll of Lhe sample wan arljufUiHl wll.li Hi\0\\ l.o pll (). fl l:i) 1.2 borore vise. The test was peif or intMi wlUi iiS ImHod on I )H> followinq senaory evaluation scorcj! coni, inl i i) iioint) and I) I iil.at.hionn addit:. Ion i 3 points. Table 'J nhowH ilm lomiltH. I OOVfi I

Tub hi 4
Kokumi intensity
CimnQiiti'atlon I'l/cll)
rwinip 1 ()
Tnal (I |)i (II 111)
Xnltlnl, and middle tasLo
0 "
AftortfliU «
0
Colli. I. o.I
TlilaKiiwHs, H the liiipi iivonuHit of thci body triate and the reducd.ldn in I lu< lut i in I ante, and reaults in a sweet taste close to suci:o3e. [OOiM] Table 6

I I I AI'M
l'iiht;i I) 1 Y-Glu-Cy(i-G.l.j;
Y-CUi-Va 1-Giy
Y-Glu-Val-Gly
y-Glu-Ahu-Giy
V-Glu~Abu-Gly

ConcBntiat. Ion (q/dJ )
0.05 0.0001
0.001
0.001
O.Ol

Taste Body tan to
0
1.0 0.6
1.3
0. 7
1.5

Hit tor
tnato
0
-0^5 -0.2
-0.5
-0.1
-0.1

TaHtn |nnf 1 In
Sweet tnato J B wlioUy enhanced. Sweet t«Hto iHiiin I iiiiic) In aftertaste .is jiupprnaflod and wliarp/ieHS la improvod.
.Swaot tasto iBiiuilnlncj in aftertaste is
_8up|Ji-oo8ed and HliarjMiasB _L8 improyedj;^
Sweot tflsto in onhancod in ini tial t£i."jte and
middle taste. .':iwoot tasto remaining in
aftertaste iu HiippieBHed.
Sweot tnBto in tinhfliiaed in init.ial taste and middle taste, .'iweot l:aata lemnining in affcortnato iiH Duppinsaod.

[0085] Table 7

Sample
iii) aucralose Concentration (g/dl) Ta3te Taste profile

Body taste Bitter taste

Control - 0 0 -
Y-Glu-Cys-Gly 0.06 1.0 0 Sweet taste with growth is enhanced in middle taste.
Y-Glu-Val-Gly 0.0001 0.7 -0.1 Outstanding sweat taste is harmonized in initial taste and aftertaste.
Y-Glu-Val-Gly 0.001 1.3 -0.2 Outstanding sweot tnate is harmonized in initial taste and aftertaste.
Y-G:iu-Abu-G].y 0.001 0.5 1.0 -0. 1 -0. 1 .Swool. tnsl-.e in onlinncod In Initial taste and mid'llo tnntn. I'.WOHI. (.asto remaining in «f l.iiii I nBti« Is tnnipiiMMied . HWBMl 1 awl H IB «nlirtni,'fi(l In initial taste and middle taBte. Ilwoiit taste remaining in aftertaste is suppressed.
y-Glu-Abu-Gly 0.01

[0086]
38

Table 8

.'3iiin|jJ,(.' Concentration (g/dl) TaHte Taotp piDf ,i l«i
i, V) A<;e-K
Body
taste Bitter taste

('Diit. rol
V-CJ ii-Cyii-Cily 0.05 0 1.0 1 . -0.6 Sweet taste is oiiliaiicud in middle taste to aftertaste.
Y-Glu-Val-Gly 0.0001 0.7 -0.6 Swoot taste with tjrowth is enhanced in middle taste.
y-Glu-Va.l-Gly 0.001 1. 2 -1.2 Sweet taste with growth is enhanced in middle taste.
y-Glii-Abu-Gly y- t! 1 u-Al>il-i.',.l.y 0.003. 0.01 0.5 1. 1 -0.6 -1.2 Sweet taste is enhanced in initial taste and middle taste. Sweet taste remaining in aftertaste is suppressed. Sweot taste is oniinncod in initial taste and middl(( t.«ntH, tlwi'icit taatci lomaining in nftoilnHtK in mnnH HHinid .
|00«7]
I I'lHainp.l t> 2] Activil.y of compound to ha uHf)inta were defined, respectively. The highar Hcort) I hftn 0 point iiiuanH l;hat the sample has a body taste-improvin<) nl'f«ol:.
On the other hand, the sensory evaluation fov l.ho improvement ot I: ho h it lor t nst ownn perf nrmed wi th n=l 2 on I.ho baa I H of the fo U,owing criteria: a control (obtained with adding diBtlilml water only and without adding glutathione and/or the peptide oC the present invention) was defined as 0 point, and by using tlio cont.rol as a standard, weak: -2 points; slightly weak: -1 point? olightly strong; I point, and strong: 2 points were defined, respecl.ively, The lower score than 0 point means that the sample has a bittarneBS-improving effect.
40

As a result of the evaluation, activities were widely exhibited at the above-mentioned addition concentrations . Tables 9 to 12 each show the results at typical concentrations . As cleai: f rom the results shown in Tables 9 to 12, it was confirmed that the peptides of the present invention each has a body taste-improving effect at a lower concentration than glutathione. Further, it is also confirmed that thepeptideof the present invention has a bitterness-improving effect. In general, the addition of the peptides provides the improvement of the body taste and the reduction in the bitter taste, and results in a sweet taste close to sucrose. [0091] Table 9

Sample ii) Concentration
(g/di) 'i'«u(.e I'd 111: II in III i In
APM:Ace-K = B:2
Body ta.sie Bitter taste

Control - 0 0 -
Y-Glu-Cys-Gly 0.05 1.0 -0.3 Sweet taste is wholly enhanced.
y-Glu-Val-Gly 0.0001 0.6 -0.3 Sweettastowlth growth la enhanced inroiddlo taste. Sweet taata i:omalning in aftertaste is auppresaad and sharpness is improved.
Y-Glu-Val-Gly 0.001 1.3 -0.7 Sweet taste remaintncj in aftertaste is suppressed and sharpness is improved.
y-Glu-Abu-Gly 0.001 0.6 -0.3 Sweet taste is enhanced in initial taste and middle taste. Sweet taste remaining in aftertaste is suppressed.
Y-Glu-Abu-Gly 0.01 1.3 -0.7 Sweet taste ia enhanced in initial taste and middle taste. Sweet taste remaining in aftertaste is suppressed.
[0092] Table 10

Sample iii)
sucralosetAce-
K=7:3 Concentration (g/dl) Taste Taste profile

Body taste Bitter taste

Control - 0 0 -
Y~Glu-CyB-Gly o.os 1 .0 -0.3 Sweet taste in enhanced in middle taste,
Y-Giu-Vai-Gly 0.0001 0.7 -0.3 Harmuiiy ia wholly pre.'jont. Heavy taoLc in suppressed and ahnrgneaa is improved.
Y-Glu-Val-Gly 0.001 1.3 -0.7 Haimony is wholly piosent. Heavy taste in suppreased and sharpness is improved.
Y~Glu-Abu-Gly 0.001 0.5 -0.3 Sweet taste is enhanced in initial taste and middle taste. Sweet taste remaining in aftertaste is suppressed.
Y-Glu-Abu-Gly 0.01 1.1 -0.7 Sweet taste is enhanced in initial taste and middle taste. Sweet taste remaining in aftertaste is suppressed.
[0093;
41

T,:ililc 1 i
!l.l||l|l I I' I \' I
AI'M 1 IJI|IT,I I M)l(':
111
I'niil HI I
y lllu I'yii- (l.ly
Y-t;lu-Va:i -G3y
Y Ulii' Vrt I ■•-CPI y Y-(:ilu-Aliu-(}.l y
Y lilu- AlMi-dl.y

Coiu'iiiil. (il/il 1 1 ill; ion T.lHl t>
llody Hi ht.oi
t unlet tasto
0. (l!i .,.,, , .1_.0_ _ -0. .1
0.0001 0.6 -0.2
0.00 1 1. 3 -O.'l
0.00.1 0.6 -0. 1
0.0 I 1.3 -O.l

Tftht !• |ii (i( 1 I <>
Mwaol t ««»,«» In Hiilifliii-tMl in miildiif tflHt.i^. Harmony in wholly ptoiioiit ami sltoiig taal.H
is achlnyorl.
Harmoiiy iBwhuiry pi ntieint rti\(J «l:i:onq tflHlB
t_« flt!ll I BVUll .
Hairmony is piBHBtil , tlwwut'. tftiiLe is enhanced
III inllinl l«Bt.n nnii (iitdiMa Inote.
llnrmony lM|)i«(((inl . .'iwufll, inntulnonhanceri
in inlilnl innln nnd iniclilto laste.

[0094] Table 12

aample v) APM:aucralose: Ac:e-K--2-. 1 : 1 Control ConoQntration (y/dl) Taste TaHte proffilB

Body taste Bitter taste

- 0 0 -
y-GIii-Ci^s-Gly Y-Glu-Val-GIy O.OS 1.2 -0.3 Sweet taBte In wholly enhanced, Harmony IB wholly protjent. rioavy taste Iti suppreased and iihnrpnaas In aftartaBte in improved.

0.0001 0.6 -0.3

V-Glu-Val-Gly 0.001 1.3 -0.7 Harmony la wholly (iriiMont. Heavy taste in supprBBBod and HharpnuHO In aftertaste in improved.
Sweet taste in enhancod in initial taste and middle taate. flWHOt (.nsto remaining in aftertaste is suppressed.
y-Glu-Abu-Giy 0.001 0.6 -0.4

Y-Glu-Abu-Gly 0.01 1.3 -0.8 Sweet taste is enhanced in initial taste and middle taste. Sweet taste remaining in aftertaste is suppressed.
[Example 3] Activity of compound to be used in present invention on sweet taste (III)
A peptide in which a calcium receptor-activating effect and a kokumi-imparting activity were found, «tul alnacnlcot which was known to have a calcium receptor-activating effect were examined for their activities (improvement of body taste and improvement of bitterness) on typical high intensity sweeteners by a quantitative sensory evaluation test. [0095]
The quantitative sensory evaluation test was performed as follows . As for standard solutions of each sweeteners , the following compounds were used. The following standard solutions have been
42

adjusted so as to have almost the same degree of sweetness as each
other.
i) Sucrose (5 g/dl)
ii) APM (0.02 5 g/dl)
iii) Sucralose (0.008 g/dl)
iv) Ace-K (0.025 g/dl)
It should be noted that an addition effect of the samples on each of the high intensity sweeteners was evaluated with reference to sucrose. 10096]
To distilled water containing each HwootenarH, v-^"'l-i'-Val-Gly oi cinacalcet was mixed as a sample at concnnl « at I emu of 0.0001 to 1 g/dj , respectively, and then each degreo of tint I mpi uvumonl- of t hn r)t)dy taBi.e and the improvement of iho lit t t ni nBMii on I lui Hweet in III (• w a H mo a B U r e d .
AB liof a sample which has stronger acidic prupnrLleH than the Htandard solution not containing the sample when the uample is dlHsolved, pH of the sample was adjusted with NaOH to nchleve a range of pH±0.2 with respect to the pH of the standard sol lit J on before use?. [0097]
The sensory evaluation for the improvement of the body taste was performed with n = 12 on the basis of the following criteria: a control (obtained with adding distilled water only and without adding a sample) was defined as 0 point, and by using the control as a standard, weak: -2 points; slightly weak: -1 point; slightly strongs 1 point, and strongs 2 points were defined, respectively. The higher score than 0 point means that the sample has a body taste-improving effect.
On the other hand, the sensory evaluation for the improvement of the bitter taste was per formed with n= 12 on the basis of the following criteria: a control (obtained with adding distilled water only and without adding a sample) was defined as 0 point, and by using the control as a standard, weak: -2 points; slightly weak: -1 point; slightly strong: 1 point, and strongs 2 points were defined.
43

respectively. The lowei; iscnre than 0 point, VIKUIMH t hal the sample has a bitterness-improving effect.
As a result of the evaluation, activities were wi dely exhibited at the above-mentioned addition concentrations. Tables .13 to 15 each show the results at typical concentrations. As clea.r from the results shown in Tables 13 to 15, it is confirmed that y-Glu-Val-Gly and cinacalcet each has a body taste-improving effect at almost the same concentration. Further, it is also confirmed that the sample of the present invention has a bitterness-improving effect. In general, the addition of the sample provides the improvement of the body taste and the reduction in the bitter taste, and results in sweet taste close to sucrose. [0098] Table 13

SampJ.Q 11) APM Concentration (q/cll) Taste Taota proJ^ilo

Body
taste Bitter taste

Conti.'oJ - 0 0 -
Y-t:;l.u-Val.-G.ly Y-Glu-Va.l.-Gly O.OOOl
o.oox 0.5 -0.2 Sweat taate cmnniiiing in nftertaate is sugi>rBftBed and H1I«HJIIOH8 is improved. Sweet taate remaining in aftertaste is suppreased and sliarpneas is improved.

1.3 -0.5

Cinacalcet 0.0001 0.8 -0.2 Sweet taste remaining in aftertaste is suppressed. Growth is present in middle taste to aftertaste.
CinacalcQt 0.001 1.5 -0.5 Sweet taste remaining in aftertaste is suppressed. Growth ia present in middle taste to aftertaste.

I 0 0')9 ] Table 14
tl«iii|:il ti
.III) HIIIU (1 I OHO
I 'Mill n> I
, (11 \i Vrtl -<11 y
\ Ulu Vnl -(.'Ay
CI wac.ulc.el
el iiaiiu loot

Coiicfint.i'at: ion |c|/dl)
0.0001 0 . I) 0 1
0.0001 0.00 1

Taate Dody
tflBtl!
"n'^
0.7 1 .3
0.5
1.1

Bitter _taste
"n""
-0. I -0.2
-0. 1
-0.2

TaHl H |ir()f 1 I M
OuLBirtnditu) Mw«m IflBi.n in harmonized in
Ijlll. ifll lant.n ali, (illl-Val "(My
/ i;iii.-Val -ci y
1' 1 IHIl'd ll.'Hl Conaentration (g/dl) Taste Taste profile

Body
taste Bitter taste

- 0 0 -

0 . 0 0 0 J
I) . 0 0 i O.OOD) () .(101 0.6
1 . 3 O.S 1.1 -0.3 -0,7 Sweet t n«l H wl l.li (jrowth ia anhanced in middle Iflflln. l!wt>«1 1 rtHi n 1 nnin 1 111 fii| in fl£tertaatt> In fuiinnHMHHil (Hilt Hiirt 1 i^itinna In improved. iJwetM (Hiitn imimiiiliig In nftMitanto ig ou[ipi HHSail /tnd Hlmi jiiiBdti l« Iniprbvod. !iwoo( 1 viHVMwl 1 h i|i nwt h I « nitliniiowd InrnJildlu 1.«Hi 11 t illil- V/il-Ciy
Y-CUu-Val-Gly
Cinacalcet
Cinacalcet

Conuontration
(q/dl)
0 . 0 0 0 I. 0.00 1
0.0001 0.001

Taste
Body taste
jO
0.7
1.3
0.8
1.5

Bitter taste
-0.3 -0.7
-0.1
-0.3

Tnnl.o pi of i In
HninKiny ia wliully PIBBBIII . Moavy taste in HuppiuMHed ami Hlinipnotu) Iti lmpiove£l. llaniKJiiy In wholly piiiuoat.. Heavy La.'sta ir, aupprasBed _and aliai{l,|ii_esji_ ia improved
Sweet taste with thickness and growth is enhanced in iiiUldlo taste to aftertaste.
Sweet i.aata with thickness and growth is enhanced in mlildle taste to aftertaste.

47

[0107] Table 11

Sample iv) APMi aucralose--
1:1 Concentration (g/dl) TflHte tiody" taste Bit¥er taste 'rnii( (1 |ii,(ir i In
Control - 0 0 -
Y-Glu-Val-Gly 0.0001 0.6 -0.2 Harmony is wholly prosont and strong taste is achieved.
Y-Glu-Val-Gly 0.001 1.3 -0.4 Harmony is wholly pi oiient and strong taste is achiaved.
Cinacalcet 0.0001 0.5 -0.2 Sweet taste with thickness and growth is enhancod in middle taste.
Cinacalcet 0.001 1.1 -0.2 Sweet taste with thickness and growth is enhanced in middle taste.
[0108] Table 19

Sample v) APM:aucralose: Aca-K=2:l:l Concentration (g/dl) Taste Taste profile

Body
taste Bitter taste

Control - 0 0 -
y-Glu-Val-Gly 0.0001 0.6 -0.-3 Harmony is wholly present. Heavy taste is suppressed and sharpness in aftertaste ia improved.
V-Cilu-Vfl]-Gly C) nacfllcet 0.001 0.0001 1.3 -0.7 Harmony is wholly present. Heavy taste in suppressed and iiharpnoas in aftertaste ia imji^i'oved.

0.5 -0.2 Harmony and richness are present in middle tflslB to aftoitflHta.
Cinacalcet 0.001 1.1 -0.4 Harmony and richnoas are present in middle taste to aftertaste.
[Example 5] Activity of the compound to be used in present invention in drink yogurt containing high intensity sweetener
A peptide in which a calcium receptor-activating effect and a kokumi-imparting activity were found, and cinacalcet which was known to have a calcium receptor-activating affncM wru'o examined l;or their activities (improvement of body t.OBt.o cirul Improvomont of bitter tasl;e) in a drink yogurt containing n 1 ypiiurl hitjh intensity HWHetener by a quantitative sensory evaiuation IOMI:. I 0 109 I
The quantitative sensory evaluation t.eBt W«H performed as follows. As for a drink yogurt standard solution, the following compounds were used. The following standard aoJuiilons have been adjusted so as to have almost the same degree of Hweetness as each other.
48

i) Sucrose (5 g/dl) :ii) APM (0.02 5 g/dl)
It should be noted that an addition effect ol the sample on each of the high intensity sweeteners was evaluated with reference to sucrose. [0110]
To a drink yogurt containing each sweeteners, y-'jl'^i-Val-Gly or cinacalcet was mixed as a sample at concentrations 0.0001 to 1 g/dl, respectively, and then each degree of the improvement of the body taste and the improvement of the bitter taste on the sweet taste was measured.
As for a sample which has stronger acidic properties than the standard solution not containing the sample when the sample is dissolved, pH of the sample was adjusted with NaOH to acliieve a range of pH±0.2 with respect to the pH of tlie Htfindrtrd nolution before use. [0111]
The sensory evaluation for the improvement of the body taste was performed with n=12 on the basis of the following criteria: a control (obtained with adding distilled water only and without adding the sample) was defined as 0 point, and by using the control as a standard, weak: -2 points; slightly weak: -1 point; slightly strong: 1 point, and strong: 2 points were defined, respectively. The higher score than 0 point means that the sample has a body taste-improving effect.
On the other hand, the sensory evaluation for the improvement ofthebitter taste was per formed with n=l 2 on the basis of the following criteria: a control (obtained with adding distilled water only and without adding the sample) was defined as 0 point, and by using the control as a standard, weak: -2 points; slightly weak: -1 point; slightly strong: 1 point, and strong: 2 points wore defined, respectively. The lower score than 0 point means that the sample has a bitterness-improving effect.
As a result of the evaluation, activities were widely exhibited
49

at l,hc abovo-mentioned addition concentrat i OUH . TJIIJIC 20 nhows the remiltH at: l.yplcal. concentrations. As clem I lom I lu> ntHults shown in 'I'able 20, It is confirmed that Y-Glu-VaJ.-Ci 1 y finil iH ncicfi lc(»l each )ia,'i a iHKly t.ciH l:e-i inproving effect at almoHt. tho Mniim conciintrntion. I'lii t hur, it IB also confirmed that the sample of Lho pi oHtuit invention hail a b i tlornoHH-improving effect. In gonern I , l:lin fujd 11, ion of the Hample provides the improvement of the body taate and the reduction in the bitter taste, and results in a sweet taste close i.o sucrose.

[ 0112 ] Table 2 0
llnmplo i I ) AI'M
Ccinurol _
Y-Glu-Val-Gly
Y-Glu-Val-Gly
Cinacalcet Cinacalcet

Conceiiti"ation
0.0001 0.001
0.0001 0.001

TaBte
Body taste
0.5
1.4
0.5
1.6

Bitter taste
-0.2
-0.5
-0.2
-0.1

Taste prof I \(Sweat Laaio lomnlniritj In aftertaste is
suppioasod.
Growth is pr(30Biit in middle Fas'te, Sweet
taateremaininij In alJ^grtasteia suppressed.
Sweet taste remaining in aftertaste is ^u^presaed.
Thickneaa is pi»inont. Swoet tasto remainlnfj in aftHrtnate ia Biinpr«i!L''!L'i-

Industrial Applicability [0113]
The use of the sweetener, in particular, the high intensity sweetener such as aspartame in combination with the compound having a calcium receptor-activating effect, preferably the amino acid or the peptide, improves the body taste or the bitter taste of the high intensity sweetener such as aspartame. The sweetener of the present invention which has the improved body taste and bitter taste of the sweet taste can be widely utilized for genera I foods Huch as beverages and water ices.
Further, the compound having a calcium receptor-activating effect can be widely utilized as an additive agent capable of improving the body taste and the bitter taste of the sweetener, in particular, the high intensity sweetener.

WE CLAIM:
1, A sweetener which comprises a sweet substance and a compound
having calcium receptor-activating effect.
2. The sweetener according to claim 1, wherein the compound
comprises one or more kinds of amino acid oi: peptide selected from
the group consisting of y-Glu-X-Gly where X represents an amino acid
or an amino acid derivative, y-Glu-Val-Y where Y represents an amino
acid or an amino acid derivative, y-Glu-Ala, y-Giu-Gly, y-Glu-Cys,
Y-Glu-Met, Y-Glu-Thr, y-Glu-Val, y-Glu-Orn, Asp-Gly, Cys-Gly,
Cys-Met, Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, y-Glu-Met(0),
Y-Glu-Y-Glu-Val, Y-Glu-Val-NH2, y-Glu-Val-o1, y-Glu-Ser, y-^lu-Tau,
Y-(M ii-Cya(S-Me) (0) , Y~Gl'J~T-'eu, Y-Glu-ll<", y-'»'^'~*^-''"'> ^^'^id
y~Glu~Cya(S-Me).
J. 'I'ho HweeUonot: according to claim 2, whoroln X reproHonts CyHliiNO), Cys (S-al lyl) , Gly, Cys(S-Me), Abu, t.-],ou, CIB, Alb, Pen cii Htif, and Y represents Gly, Val, Glu, LyH, l'lu», Moi , Pro, Arg, Asp, Met, Thr, His, Orn, Asn, Cys, Gin, GlyA or LacA.
4 . The sweetener according to claim 2 or 3 , wherein the compound
i ,'i :u.'Iect:o(i from y-Glu-Val-Gl y and y-Gl \i-Abu-Gl y .
5. The sweetener according to any one of claimH 1 to 4, wherein the sweet substance comprises one or more kinds of substance selected from the group consisting of aspartame, sucralose, acesulfame K, neotame,
N-[N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-a-aspartyl]-L-phenylalanine 1-raethyl ester, saccharin, stevia, glycyrrhizin, thaumatin and monellin.
6 . Anadditiveagenttobeaddedtoasweetsubstance for improving a body taste of the sweet substance, which comprises a compound having

calcium receptor-activating effect.
7.An additive agent to be added to a sweet substance for improving a bitter taste of the sweet substance, which comprises a compound having calcium receptor-activating effect.