METHOD FOR CONTROLLING NEUROTRANSMITTER USING CUCURBIT[N]URIL, AND USE THEREOF

The present invention refers to release of neurotransmitter reel number plower method using relative cooker and relates to uses of, an empty cell membrane fusion protein role as intermediate relative cookie SNARE interacts neurotransmitter release of coating neurotransmission stopping system including composite number cookie relative reel and SNARE protein [...] substrate.

The present invention is an empty cell membranes victims of the important role it is the beginning molecular sieve cookie relative SNARE protein fusion proteins interact with SNARE membrane fusion of deep-seated to interfere each other while a complex number billion to find that release of neurotransmitter release of neurotransmitter plower by the present invention to construct a system number is more than 1.70.

Generally nerve cells communicate with one another and exchanges signals with a respective bolted when cells where it is, at the end of the synapse in the neuronal neurotransmitter storing pocket gun [nang fused to cell membranes which are activated neurotransmitter. At this time cell membranes that are most significant role (SNARE) protein fusion protein in user in liquid is sprayed directly [...] neurotransmitter protein. (Synaptobrevin provided 2) is placed on the bag side marker gun [nang [...] protein on the membrane side marker v provided SNARE t provided SNARE consisting of (Syntaxin-a 1 and SNAP-a 25). [...] alpha - helix bundle protein to form a complex which mutually twisted coil force required membrane fusion occurs.

However the present invention it is the beginning molecular sieve (CB) is the victims of the cookie [...] interacts protein relative reel function the present invention by finding out that enters the inner composite inhibiting the arrears of work.

Relative cooker (CB) reel macrocyclic molecules with hydrophobic cavity inside which is rigid and has a buckle charge CB [...] melting in water is a carbonyl group (carbonyl) owner molecules are present in the inlet cavity has a strong hydrophobic interaction based on stable owner - - ion dipole interactions guests formation (host - guest) complex protein interaction neurotransmitter release of an empty one cookie [...] relative elastic knew the number.

[N] of the present invention relative rotation [...] cookie that proteins interact in a sachet bag labeled protein [...] effectively neurotransmission by stopping such as wrinkles or aging skin cosmetic composition or alternative number [...] release of neurotransmitter billion for composition can be used in number.

[N] or aging skin wrinkle cosmetic composition of the present invention relative cooker which reel, cookie relative to release of neurotransmitter [...] number through an empty SNARE proteins can improve skin such as a wrinkle or aging.

Heat or aging skin wrinkle cosmetic composition of the present invention prior to as an empty cookie relative to release of neurotransmitter [...] proteins neurotransmission billion billion or aging skin wrinkles number as a number communicated to the number for use to improve [...] hereinafter can be replaced.

The number existing [...] taken as does not use dangerous or life-threatening toxicity but has enough, cooker relative rotation of the small size while the efficacy of compound as [...] with, [...] side effects and can be reflected to the input port.

In the embodiment according to formula 1 of the present invention for rotation relative cookies in one cosmetic composition can be represented.

[Formula 1]

[In said formula 1,

X is O, NH or S and,

A₁ And A₂ Are each independently H, - OR, - SR, - NHR, - COOH, - O (CH₂ )_a R or - O (CH₂ )_a SR and, said a is integer number of 1 to 5, said R is H, halogen, C1 - C30 alkyl, C2 - C30 alkenyl, C2 - C30 alkynyl, C2 - C30 carbonyl alkyl, C3 - C30 cycloalkyl, C2 - C30 cycloalkyl, C6 - C30 aryl, C6 - C30 aryl C1 - C30 alkyl, C2 - C30 heteroaryl or heteroaryl C1 - C30 alkyl C2 - C30 and;

N is an integer from 4 to 14.]

In the embodiment according to said formula 1 of the present invention in which X is one cosmetic composition preferably O, A₁ And A₂ Are each independently H, - OR, - O (CH₂ )_a R or - O (CH₂ )_a SR and, said a is integer number of 1 to 5, said R is H, C1 - C30 alkyl or C2 - C30 and [...]; n is 6 to 10 flickering disclosed.

In the embodiment according to said formula 1 of the present invention in which X is preferably one O, A₁ And A₂ Are each independently H or - OR and, said R is H or C1 - C30 may be alkyl disclosed.

The present invention a "alkyl", "alkoxy" and other "alkyl" which contains both straight or grinding form portion including substituents, preferably 1 to 30 carbon atoms 1 to 20, more preferably 1 to 10 carbon atoms has.

The present invention the terms "alkenyl" alone or as part of a 2 to 30 carbon atoms 1 and another at least one carbon to carbon double bonds containing linear, branched or cyclic hydrocarbon radical big. In a more preferred alkenyl radicals alkenyl radical having 2 to about 20 carbon atoms are disclosed. The most preferred lower alkenyl radicals having 2 to about 10 carbon atoms radical are disclosed. In addition can be substituted at any available point of attachment situated neel the [khey which it will know it crawls. Examples of alkenyl radicals ethenyl, propenyl, allyl, propenyl, butenyl and 4 - methyl neel vice-reel multiple myelomas are included. Terms alkenyl and lower alkenyl cis and trans orientation, or alternatively, comprise a radical Z E and orientation.

The term "alkynyl" as part of the present invention alone or another a 2 to 30 carbon atoms containing at least one carbon to carbon triple bond 1 and linear, branched or cyclic hydrocarbon radical big. In a more preferred alkynyl radicals lower alkynyl radical having 2 to about 20 carbon atoms are disclosed. Lower alkynyl radical having 2 to about 10 carbon atoms most preferred are those are disclosed. Examples of such radicals propargyl, like father [thin one multiple myelomas are included. In addition the alkynyl can be substituted at any available point of attachment situated.

The present invention is a "cycloalkyl" having 3 to 30 carbon atoms (monocyclic) or polycyclic (multicyclic) means a difference between magnetic and non-ring type, the present invention is a "cycloalkyl" carbon atoms and nitrogen, phosphorus, 1 to 5 heteroatoms selected from oxygen and sulfur or a substituted or unsubstituted non-aromatic 3 to 15 won two substituted cycloalkyl and cycloalkyl is a ring to the present invention 3 to 10 won ring, preferably 5 to 7 won can be ring.

The present invention carbon atoms "cycloalkyl" is a nitrogen, phosphorus, oxygen and sulfur substituted 1 to 5 heteroatoms selected from two or a substituted or unsubstituted non-aromatic 3 to 15 won ring radical dispersing medium, is fused or cycloalkyl radicals, or otherwise capable of bridge ring members spiro ring line containing the case type, bicyclic or tricyclic ring and may be an ellipsometer, the heterocyclic ring radical nitrogen, phosphorus, carbon, oxygen or sulfur atoms can be oxidized when various oxidation state according. In addition, when nitrogen atoms can be dialkylamide according to 4.

In addition the present invention "aryl" is a number one hydrogen derived from aromatic hydrocarbons by volatile organic radicals, preferably two to 4 each ring 7, preferably 5 or 6 ring atoms including single or fused ring orgin of which, it will be biting, this type comprising a plurality of single which is connected to. In particular for example phenyl, naphthyl, biphenyl, [...], indenyl (indenyl), fluorenyl bromic acid, is not limited.

The present invention as a "heteroaryl" is an aromatic ring backbone atoms B, N, O, S, P (=O), selected from 1 to 4 hetero atoms of Si and P, meaning that the aryl group is an aromatic ring backbone atoms remaining carbon may be used, 5 to 6 won only annularity heteroaryl, and at least one va and condensed polycyclic hetero and it will be biting, this, partially saturated disapproval. In addition, a variety of one or more heteroatoms in the present invention it will be biting it will be biting, this single form comprises a unit which is connected.

1 To 10% by weight relative rotation of the present invention cosmetic composition can be included with the one in the embodiment according to said cooker.

One cosmetic composition of the present invention in the embodiment according to number type suspension, emulsion, paste, gel, cream, lotion, powder, soap, number surfactants containing cleansing, oil, powder foundation, emulsion foundation, wax foundation or spray be a.

In the embodiment according to of the present invention cosmetic composition of the present invention cosmetic compositions on the reel outside relative one cookie can be antiseptic number, moisturizing number, stabilizing number, emulsion number, number since, liquid crystal formula of number, pigment and distribution tube further comprises one or more selected from addition number can be conventional. The, adding said number is not limited and if the material commonly used in the art, specific one example include least one alcohol, alcohol and hemp cloth [hey neel alcohol fat (fatty) alcohol and bis 15/methyl ethyl dimethyl silane such as triacetyl easy pin, pin-in-not-in-- 10/15 /, /-in--3 polyglycerol, d maul nose glow-in-/, /-in--in--in-, cycle/[...] d maul nose glow, cycle-in-[...] /, / tree methyl thread rock sicily kate [...] cycle, cycle-in-/ pen hit ceylon from, pin-in--12 not pen hit ceylon from 1 /, // pen hit ceylon from cycle-in-least-in-, cycle-in--in-pen hit ceylon from / /, / silicon polymer in NaCl-in-polymer or the like-in-stabilized number; number [...] based cationic, anionic surface active number, number amphoteric surfactants, non-ionic surfactants can be mixed emulsions in NaCl number number, preferably a poly glyceryl -4 relay [thu capric acid/capric acid rate, capric acid/capric acid glyceryl poly -5 relay [thu rate, capric acid/capric acid glyceryl poly -6 relay [thu rate, capric acid/capric acid glyceryl poly -7 relay [thu rate, capric acid/capric acid glyceryl poly -8 relay [thu rate, capric acid/capric acid glyceryl poly -9 relay [thu rate, capric acid/capric acid glyceryl poly -10 relay [thu rate, poly glyceryl -4 rate capric acid, capric acid rate glyceryl -5 poly, poly glyceryl -6 capric acid rate, poly glyceryl -7 rate capric acid, capric acid rate glyceryl -8 poly, poly glyceryl -9 rate capric acid, capric acid rate glyceryl -10 poly, poly glyceride reel -4 laurate, poly -5 glyceryl laurate, glyceryl laurate -6 poly, poly -7 glyceryl laurate, glyceryl laurate -8 poly, poly -9 glyceryl laurate, glyceryl laurate -10 poly, poly glyceryl -6 coco polycarbonate, poly glyceryl -7 coco polycarbonate, poly glyceryl -8 coco polycarbonate, poly glyceryl -9 coco polycarbonate, poly glyceryl -10 coco polycarbonate, poly glyceryl -11 coco polycarbonate, poly glyceryl -12 coco polycarbonate, poly glyceryl -6 pre-stay [thu, poly glyceryl -7 pre-stay [thu, poly glyceryl -8 pre-stay [thu, poly glyceryl -9 pre-stay [thu, poly glyceryl -10 pre-stay [thu, poly glyceryl -11 pre-stay [thu, poly glyceryl -12 pre-stay [thu, poly oleyl glyceryl -10 polycarbonate, poly oleyl glyceryl -11 polycarbonate, poly oleyl glyceryl -12 polycarbonate, poly -10 glyceryl stearate, glyceryl poly -11 stearate, glyceryl stearate -12 poly, glyceryl fatty acid ester-based surfactants such as glyceryl poly -6 reacting directly with writing three reel number s group composed as a high pressure liquid coolant or poly fatty acid number as well as the seller using commercially available vertical transfer are disclosed. Said cosmetic composition has an appropriate viscosity number permanently imparting feeling after use and improve the stability of number into a carbomer, carbonate pole, gelatin, a thickening, natural cellulose, high cells, but not the limited to selected from methyl cellulose and the like. Liquid crystal of said liquid crystal is number formula to an increase in rigidity, so that a tightly hold connecting fence that helps maintain properties play a role in, a factor, bis-hydroxyethyl bis tax [thil at end amide, isocyanate cholesterol stearate, cholesterol oleyl light, cholesterol stearate, lecithin, (one example, ceramide 3, ceramide 6) ceramide current etc. but not the limited to.

In addition said pigment extender pigment, white pigment, coloring pigment, a pearlescent pigment, metallic powder, organic powder and the like, said extender pigment include talc, mica, kaolin, calcium carbonate, mica, talc, kaolin, alumina, barium silicate, number it rises, light, radiating far-infrared rays, magnesium carbonate, barium sulfate and capable of, white pigment include titanium oxide, zinc oxide and capable of, coloring pigment include [...], iron sulfate, iron black it buys, chromium oxide, [...], lawful interception, and carbon black and capable of, pearlescent pigment include titanium dioxide, titanium mica, titanium oxide coated mica titanium it buys season and, silica, [thin jade site, and where at the time of it knows the [tu and capable of ferrocene, metallic powder include gold, is, copper, palladium, platinum and capable of, organic powder include polymethylmethacrylate, nylon, cellulose, starch capable of disclosed. In addition typically publicly known cosmetic and R2 natural, inorganic, and an organic pigment can be both, said natural pigment include Gardenia yellow, blue pigment blue, Gardenia yellow green, blue pigment red, monascus the dye, monascus sulfur dye, safflower [...], drug earth dye, and consequently boosts, lock dye, quantity dye, pigment grapes chili pepper, [...] dye, Elderberry pigment, blueberry pigment, pigment [phwu car, the [khya it will carry on shoulder dye, pigment enemy nothing right, feeling dye, phase dye, riboflavin, beta carotene, drugs dye, pigment [...], cone red dye, bit red dye, inside inside wristlet, anthocyanin, pico cyan, it is not cauchy blood, it will bloom with the claw dye, and combinations thereof can be selected from the group consisting 1 paper, inorganic pigment include metal oxide, in particular, iron oxide (red, black, yellow, brown), titanium dioxide, zinc oxide, chromium oxide, bismuth oxychloride, aluminum oxide, zirconium oxide, cobalt oxide, cerium oxide, nickel oxide, calcium hydroxide, iron hydroxide, aluminum hydroxide, chromium hydroxide, magnesium hydroxide, the lung [lik ammonium perot city it knows but the id, [...] blue, iron sulfides, manganese violet, carbon black, mica, kaolin, and combinations thereof can be selected from the group consisting 1 paper, organic pigment include indigo rake, carboxylic ketoimine rake, well-known FD&C and D&C dye series derived from rake, e.g. D&C Red 21 aluminum rake, calcium D&C Red 7 rake, directional azo, indigo-synchronized, triphenylmethane, do not write the [khwi rice field [...] dyeing such as natural or synthetic organic dye is used as the number and pivotably.

In addition the present invention refers to reel relative cooker which, relative to an empty SNARE proteins neurotransmitter release number [...] multi-adhered on neurotransmitter release pharmaceutical composition for lengthening number number [...] substrate.

SNARE proteins of the present invention pharmaceutical composition prepared by neurotransmitter release number reel neurotransmitter by including the cookie [...] relative number effectively billion spinal cord can be used effectively in the treatment and prevention of muscle related diseases or mental disorders.

A medicinal composition of the present invention in a reduction in the current method for enhancing the cookie included relative rotation [...] disadvantages attendant with the side effects while substantially reducing the size [...][...] billion number equivalent to or not less neurotransmitter release of powder can be used in proboscis related diseases or mental diseases can.

In the embodiment according to methods of the pharmaceutical composition of the present invention one eyelid proboscis related disorders, with a power supply, more cervical myotonia, stiff palpebra yearly axis, the axilla pts.wt.of, anal laceration, vaginismus, relaxation disabling use, such as migraine headache diseases, neurological and idiopathic detrusor combination, topical the market symptoms which it wins, tmjd pain disorders, diabetic neurosis, vocal cord dysfunction, sensors mounted thereon, chronic neuropathies, facial muscle hyperplasia, detrusor sphincter disorder or benign prostatic hyperplasia may be, psychiatric disorders may be tiger non saddle child, phobia, panic disorders, be a compulsive disorders or trauma stress disorders.

In the embodiment according to of the present invention to provide medicinal compositions included relative rotation 0 one cookie. 001 To be included with the 1% by weight and, in effect mirror number aspects preferably 0. 005 To 0. 1 Weight % can be included.

Suitable dosage is number number of pharmaceutical composition of the present invention method, administration scheme, patient's age, weight, -, pathological conditions, food, administration time, routes, such as sensitive and reaction speed is determined by the various prescribed for a printed wiring board can be. Pharmaceutical composition of the present invention example 1 dosage is 0. 001 - 100 Mg/kg are disclosed.

Oral or parenteral administration of the present invention pharmaceutical composition prepared can, when parenteral administration is intravenous injection, subcutaneous injection, muscle injection, abdominal implantation, such as transdermal administration can be administered.

Expression of the recombinant pharmaceutical composition has a number of the present invention can be cosmetically acceptable. A medicinal composition of the present invention contained in a cosmetically acceptable when used as species typically about number number number, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium and mineral oils and the like acid including but, limited to are not correct. In addition to the components of the present invention pharmaceutical composition is said lubricating number, wet number, number sweetener, flavor number, number emulsion, suspension number, number further comprises preserving such as can be.

In the embodiment of the present invention invention is hereinafter for person with skill in the art is provided to the pharmaceutical composition is capable according to the method, a cosmetically acceptable carrier and/or excipient about number number number number number number bath to form using conventional or otherwise prepared by elder brother anger unit dose container number bath 1308. amount to of muffler. Conventional number type the circular for example oral (positive number, number capsule, powder number), oral cavity, beneath the tongue, in the rectum, in a menopausal, intranasal, topical or parenteral (intravenous, sponge body, intramuscular, including within the subcutaneous) administration has age number type. For example, the present invention according to compounds containing starch or [...] form positive number, or sole or excipient containing capsule form number, or taste or color number counted using a mechanical suspension containing chemicals l [lik hour [lu or oral form, can be administered in the mouth or under the tongue. Liquid is suspended number number number (for example, methyl cellulose, above [theyp brush (witepsol) such as semi-synthetic triglyceride or filled oil (apricot kernel oil) on PEG-a 6 ester/capric triglyceride mixture as the mixture spinning reel [lik writing scherrer id capric acid and mixtures or PEG-a 8) about number such as dermatologically acceptable addition number encoded number with bath. In addition, in a parenterally administrable for example, intravenous, sponge body, intramuscular, subcutaneous when driving through a sterile aqueous solution which form most preferably used as scan lines, the solution is to have a isotonic with blood other materials (for example salt (salt) or mannitol, glucose monosaccharide) containing disapproval.

Preferably one of the present invention in the embodiment according to pharmaceutical composition is positive number, fill, capsule, granules, powder, acid number, number solution, patch number, using microneedle patch number or injectable percutaneous absorption can be used in form of a number.

A medicinal composition of the present invention can be represented by relative rotation cookie included in said formula 1, in which said formula 1 X is preferably O, A₁ And A₂ Are each independently H, - OR, - O (CH₂ )_a R or - O (CH₂ )_a SR and, said a is integer number of 1 to 5, said R is H, C1 - C30 alkyl or C2 - C30 and [...]; n is 6 to 10 flickering disclosed.

In the embodiment according to said formula 1 of the present invention in which X is more preferably one O, A₁ And A₂ Are each independently H or - OR and, said R is H or C1 - C30 may be alkyl disclosed.

A drug composition of the present invention release of neurotransmitter [...] number can be, cookie relative reel reel supported by the horizontal release of neurotransmitter billion cookie can be guest material relative number followed by release of neurotransmitter can be resumed again.

I.e., pharmaceutical composition of the present invention included in the cookie SNARE proteins relative rotation number and the release of neurotransmitter billion, necessary relative strong cookies that interact preferentially with the damper by introducing guest material interact with relative permeability reel causes the cookie is not received [...] protein was stopped to be released neurotransmitter released by re can be normal neurotransmission and the column.

The adamantane derivative or ferrocene derivative of the present invention may be one in the embodiment according to guest material, specifically to formula 2 or formula 3 can be represented.

[Formula 2]

[In said formula 2,

R₁ And R₂ Are each independently - H, - OR₁₁ , - NR₁₂ R₁₃ Or - N (R₁₄ )₃ To X,

R₁₁ To R₁₄ Are each independently H, halogen, C1 - C30 alkyl, C2 - C30 alkenyl, C2 - C30 alkynyl, C2 - C30 carbonyl alkyl, C3 - C30 cycloalkyl, C2 - C30 cycloalkyl, C6 - C30 aryl, C6 - C30 aryl C1 - C30 alkyl, C2 - C30 heteroaryl or heteroaryl C1 - C30 alkyl C2 - C30 and;

X is halogen.]

[Formula 3]

R₃ Is - H, - OH, - OR, - NR₁₅ R₁₆ , - N (R₁₇ )₃ X, - NH (CH₂ )_r NR₁₈ R₁₉ , - COOH or - (CH₂ )_r NR₂₀ R₂₁ And, R₁₅ To R₂₁ Independently of each other H, halogen, C1 - C30 alkyl, C2 - C30 alkenyl, C2 - C30 alkynyl, C2 - C30 carbonyl alkyl, C3 - C30 cycloalkyl, C2 - C30 cycloalkyl, C6 - C30 aryl, C6 - C30 aryl C1 - C30 alkyl, C2 - C30 heteroaryl or heteroaryl C1 - C30 and C2 - C30 alkyl; X is halogen; r is an integer 1 to 3.]

In the embodiment according to said damper of the present invention in chemical formula 3 or 2 relative cookies in one for forms strong bonds in one aspect preferably R₁ To R₃ Independently of each other OH or NH₃⁺ X^- Implementation being.

In addition of the present invention reel and guest material including kits for lengthening its release of neurotransmitter relative cooker number number [...] substrate.

The kits of the present invention using cookie reel relative to release of neurotransmitter [...] proteins which number billion, an appropriate capacity guest material necessary followed by release of neurotransmitter release of neurotransmitter is effectively number [...] again for a variety of purposes, in particular their can be useful for pharmaceutical use.

In addition the present invention refers to a method it is big expense to reel release of neurotransmitter plower number using number [...] substrate.

[N] number of rotation in the embodiment according to method of the present invention release of neurotransmitter plower SNARE proteins relative to multi-billion may be the release of neurotransmitter number, relative rotation can be represented by said formula 1 cookie.

In the embodiment according to said formula 1 of the present invention in which X is O number of release of neurotransmitter plower method, A₁ And A₂ Are each independently H, - OR, - O (CH₂ )_a R or - O (CH₂ )_a SR and, said a is integer number of 1 to 5, said R is H, C1 - C30 alkyl or C2 - C30 and [...]; n is 6 to 10 and flickering, one in the embodiment according to said formula 1 of the present invention in which X is preferably O, A₁ And A₂ Are each independently H or - OR and, said R is H or C1 - C30 may be alkyl disclosed.

Method of the present invention release of neurotransmitter plower number can have a guest material followed by release of neurotransmitter can be resume, the guest material interact preferentially with the cookie can be damper relative strong material relative (owner - guests action) prepared by the neurotransmitter release of guest reel cookie kill be disclosure material.

In the embodiment according to said formula 2 or formula 3 of the guest material of the present invention release of neurotransmitter can be display number plower method, preferably R in said formula 2 or 3₁ To R₃ Independently of each other OH or NH₃⁺ X^- Implementation being.

Hereinafter, the present invention through a in the embodiment detailed as follows. Stage, in the embodiment of the present invention is exemplified ephemeral to is, in the embodiment of the present invention to defined by contents are not correct.

[...] sulforhodamine B (SRB) protein (SNARE) manufacturing method and is nested a sachet bag (v-a Vesicle), carrying no anything Biochemistry 54 is a sachet bag (t-a vesicle), 1831 - 1840 (2015) method was number as a high pressure liquid coolant. At this time SRB (Invitrogen) concentration is 20 mm so that the v-a Vesicle carried inside, to output the first quenching the fluorescence signal are disclosed.

The present invention refers to a reel (CB[7]) a sachet [...] labeled protein relative cooker the bag when it has processed, number relates to the release of neurotransmitter, specifically SRB sachet bag just fusion of the bladder to radiate the fluorescent signals from a sachet is embedded through a small neurotransmitter can be confirmed.

[Comparison example 1] release of neurotransmitter

Each at room temperature constitutes a lipid vesicle (t-a vesicle or v-a vesicle) (lipid) concentration 0. Such as a [mwulk the tongue it gives, 0 02 mm each. Number 05 ml solution of high pressure liquid coolant therein. SRB is supported therein is a vesicle v provided Vesicle solution 0. 05 Ml solution of supported on anything not t provided Vesicle 0. 05 Ml 35 °C observed a mixing in detecting the fluorescence signal (fluorescence spectrophotometer, Cary Eclipse, Varian). At this time SRB 532 nm wavelength corresponding to the excitation of fluorescent substance is irradiated with light, a wavelength corresponding to the emission of fluorescent substance corresponding to the SRB 580 nm fluorescence signal at a substrate. SRB in high concentration is 20 mm v provided Vesicle carried inside the fluorescent signal quenching (quenching) axis so that the vesicle, and is just the fusion protein interaction by [...] v provided Vesicle t provided Vesicle quenching the fluorescence signal is recorded in the SRB (dequenching) formed transferred in the fluorescent signals while d [khwen when the other. T provided Vesicle fused each other of fluorescent signals from an increase in the v-a Vesicle and is used to prevent neurotransmitter has been confirmed.

Input reel cookie has not neurotransmitter released therefrom relative nerve can be an indirectly-delivery occurs.

Release of neurotransmitter billion number [in the embodiment 1]

HEPES (4 - (2 a-Hydroxyethyl) piperazine-a 1 a-ethanesulfonic acid, sigma aldrich) 25 mm, KCl 100 mm distilled water for 1 to pH 7. 4 High pressure liquid coolant HEPES buffer was number by a user.

[7]Anumber 1 ml HEPES buffer high pressure liquid coolant relative a cookie to the reel 1. An aqueous solution after high pressure liquid coolant by melting 163 mg 1 mm relative cookie reel number, relative permeability multi-reel aqueous solution 0. A v-a Vesicle 04 ml solution 0. 05 Ml added to each other. Here t-a vesicle solution 0. 5 Ml by adding 35 °C observed in detecting the fluorescence signal. At this time SRB have non-fluorescent signal sources, this cooker relative the number just fusion protein interacts with an empty [...] protein complex [...] billion to not making sure that the capable of neurotransmitter release.

[7] Of the present invention therefrom by number just fusion protein in combination with an empty [...] relative multi-billion billion number can be neurotransmitter release of molecules.

1800 Seconds after 0. 1% Triton X-a 100 (sigma aldrich) added thereto while quenching the fluorescence signal at a vesicle is detected address recorded in the SRB dequenching is increased while detecting the fluorescence signal has been confirmed.

SRB address increases in fluorescence signal SRB in a vesicle is a vesicle while still planetary gear set, and a vesicle judges that the original function and an indirectly-cell line as well as other SRB is released fluorescence signal due to factors of the present invention but is not empty release of neurotransmitter due to relative multi-billion number can be reliably known.

Release of neurotransmitter resume [in the embodiment 2]

1 Ml in HEPES buffer to bay [theyn adamantane 1 number produced therewith said in the embodiment 1. 10 Mm aqueous solution was 8 mg bay [theyn adamantane number by melting high pressure liquid coolant. Solution added to an empty v provided Vesicle t provided Vesicle solution relative cooker blended solution to aqueous solution 0 bay [theyn adamantane (billion number neurotransmitter release of solution). 012 Ml bay [theyn adamantane 3 equivalent amount to be added to a cooker relative reel adhesive force. 30 Minutes incubation time putting the, 35 °C observed in detecting the fluorescence signal. The SRB in fluorescent signal quenching (quenching) v provided Vesicle when multivalent, v-a Vesicle and interaction by just the fusion protein is formed t provided Vesicle [...] SRB (dequenching) d [khwen [khwen it becomes, the fluorescence signal is emitted fluorescence signal is transferred while it was been. This fiber bay [theyn this adamantane [...] protein complex by combining interacts strongly relative cooker damper located inside reel gear mounted cooker relative dissociated and identifying signals to each other can be v provided Vesicle t provided Vesicle fusion. Reel neurotransmitter release is inhibited by relative fluorescent signal from a cooker is a unit, bay [theyn adamantane neurotransmitter released by making sure that the guest molecules can be.

[Comparison example 2]

Control stopping block, in solution so that the other end 35 °C v-a SNARE protein 3 uM t provided Vesicle solution 0. 05 Ml v provided Vesicle solution on 0. 05 Ml spray solution on the other end a detachable generating electrode formed on one inner peripheral surface t provided Vesicle t-a SNARE protein interacts with the SRB occurs and not just the fusion to v-a SNARE v provided Vesicle increase has been confirmed that the fluorescent signal is not detected. Recorded in the SRB is carried inside a vesicle out therefrom v provided Vesicle exit can be sure that the fluorescent signal by dequenching does not take place.