KR20230097463A - Method for promoting myelin plasticity - Google Patents

Abstract

본 발명은 희소돌기신경교세포를 표적하는 합성수용체를 포함하는 제1 제제; 및 합성 수용체를 표적하는 합성 리간드를 포함하는 제2 제제를 포함하는 신경수초의 가소성 증진용 조성물에 관한 것이다. 인산화된 ERK1/2 발현의 향상을 유도함으로써, 신경수초의 가소성 증진을 유발하고, 이를 통해 기억력, 인지능력 및 운동능력 등 신경계 기능향상을 가져올 수 있다.The present invention relates to a first agent comprising a synthetic receptor targeting oligodendrocytes; And it relates to a composition for enhancing the plasticity of the nerve sheath, including a second agent containing a synthetic ligand targeting a synthetic receptor. By inducing improvement of phosphorylated ERK1/2 expression, it induces plasticity enhancement of nerve sheath, and through this, it is possible to improve nervous system functions such as memory, cognitive ability and motor ability.

Description

신경수초 가소성 증진 방법{Method for promoting myelin plasticity}Method for promoting myelin plasticity {Method for promoting myelin plasticity}

본 발명은 신경수초 가소성 증진방법에 관한 발명이다.The present invention relates to a method for enhancing neuromyelin plasticity.

신경조직은 신경세포와 신경교세포로 이루어져 있다. 그 중 중추신경아교세포는 별아교세포, 희소돌기신경교세포, 미세아교세포 그리고 뇌실막 세포로 크게 나눌 수 있다. 신경세포는 자극을 받아들이고 정보를 처리하여 신경충동의 형태를 전도하는 역할을 하는데 신경교세포들은 이러한 신경세포를 구조적으로 보조하거나 지지하여 신경전달 속도 및 신진대사에 관여하는 역할을 한다. 별아교세포는 아교세포 중 수가 가장 많으며 신경세포를 물리적으로 지지하는 역할을 하며 항상성과 연관이 있는 교세포이다. 희소돌기신경교세포(Oligodendrocyte)는 수초증을 형성하여 신경원의 생존과 유지에 관여한다. 수초화가 일어나는 동안 희소돌기신경교세포의 세포막은 축삭을 여러층으로 둘러싸며 하나의 희소돌기신경교세포는 돌기를 통해 수십개의 축삭을 수초화 할 수 있다. 뇌실막세포는 새로운 뉴런이나 아교세포를 형성할 수 있는 신경줄기세포로의 역할을 한다. 미세아교세포는 포식작용을 하여 뇌의 방어에 중요한 역할을 한다.Nervous tissue consists of neurons and glial cells. Among them, central glial cells can be largely divided into astrocytes, oligodendrocytes, microglia, and ependymal cells. Nerve cells receive stimuli, process information, and serve to conduct the form of nerve impulses. Glial cells structurally assist or support these nerve cells to play a role in nerve transmission speed and metabolism. Astrocytes have the largest number of glial cells, play a role in physically supporting nerve cells, and are glial cells related to homeostasis. Oligodendrocytes are involved in the survival and maintenance of neurons by forming myelin. During myelination, the cell membrane of oligodendrocytes surrounds the axon in several layers, and one oligodendrocyte can myelinate dozens of axons through projections. Ependymal cells serve as neural stem cells capable of forming new neurons or glial cells. Microglia play an important role in the defense of the brain by phagocytosis.

또한, 희소돌기신경교세포에 의해서 생성되는 수초는 포유류의 뇌의 특정한 영역에서 이루어지고 성인기까지 이루어진다. 분화에 실패하고 성숙하지 못한 세포는 세포자살이나 전구체로 남는다는 연구가 있으나 아직까지 명확한 것은 아니다. 이렇게 성숙한 희소돌기신경교세포에 의해 형성된 수초는 축색돌기를 둘러싸고 있는 동심원적으로 적층 된 막 구조이다. 말초신경계에서는 슈반세포가 미엘린을 만들지만 중추신경계에서는 희소돌기신경교세포가 역할하는데 모든 축색돌기가 수초로 덮여있는 것은 아니며, 전기적 전달에 대해 절연체를 형성함으로써, 빠른 신경전달이 이루어지도록 도와준다. 또한 인접한 두 개의 미엘린 세그먼트 사이에는 갭이 존재하는데 이를 란비에 결절이라고 하며, 신경이 전달될 때 이 결절사이를 점프하는 방식으로 진행하여 빠른 전달을 유도하기도 한다. In addition, the myelin produced by oligodendrocytes is made in a specific area of the mammalian brain and continues until adulthood. There are studies that indicate that cells that fail to differentiate and remain immature undergo apoptosis or remain as precursors, but it is not yet clear. The myelin sheath formed by these mature oligodendrocytes is a concentrically layered membrane structure surrounding the axon. In the peripheral nervous system, Schwann cells make myelin, but in the central nervous system, oligodendrocytes play a role. In addition, there is a gap between two adjacent myelin segments, which is called a Ranvier node, and when a nerve is transmitted, it proceeds in a way that jumps between these nodes to induce rapid transmission.

기존 신경세포를 광유전학 및 행동을 통해 자극하고 이에 대한 부차적인 효과로 신경수초의 가소성을 유도하는 방법은 신경세포에 대한 직접적인 자극으로 신경수초의 가소성뿐만 아니라 신경계 기능이 직접적으로 영향을 받는 등의 부차적 영향이 많아 기초 뇌과학 이외에는 활용되기 어려웠다.The method of stimulating existing nerve cells through optogenetics and behavior and inducing plasticity of nerve sheath as a secondary effect of this is a direct stimulation of nerve cells, which directly affects not only the plasticity of nerve sheath but also the function of the nervous system. It has many side effects, so it was difficult to use it for anything other than basic brain science.

한국등록특허 제10-1964875호Korean Patent Registration No. 10-1964875

본 발명은 신경수초의 가소성 증진용 조성물을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a composition for enhancing the plasticity of the nerve sheath.

본 발명은 기억력, 인지능력 및 운동능력 증진방법을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a method for improving memory, cognitive ability and motor ability.

1. 희소돌기신경교세포에 합성수용체를 발현시키는 단계; 상기 합성 수용체를 표적하는 합성 리간드를 처리하여 ERK1/2를 인산화시키는 단계를 포함하는 신경 수초의 가소성 증진 방법.1. Expressing synthetic receptors in oligodendrocytes; A method for enhancing plasticity of a nerve sheath comprising the step of phosphorylating ERK1/2 by treating a synthetic ligand targeting the synthetic receptor.

2. 위 1에 있어서, 인간을 제외한 동물에서 수행되는 것인 신경 수초의 가소성 증진 방법.2. The method for promoting plasticity of nerve myelin according to 1 above, which is performed in animals other than humans.

3. 위 1에 있어서, 상기 합성수용체는 Gq-DREADD, Gs-DREADD 또는 β-arrestin preferring DREADD로 이루어진 군으로부터 선택되는 하나 이상인, 신경 수초의 가소성 증진 방법.3. The method according to 1 above, wherein the synthetic receptor is at least one selected from the group consisting of Gq-DREADD, Gs-DREADD, or β-arrestin preferring DREADD.

4. 위 1에 있어서, 상기 합성 리간드는 클로자핀 N-옥사이드, 클로자핀 및 컴파운드-21로 이루어진 군으로부터 선택되는 하나 이상인, 신경 수초의 가소성 증진 방법.4. The method according to 1 above, wherein the synthetic ligand is at least one selected from the group consisting of clozapine N-oxide, clozapine and compound-21, the plasticity enhancement method of nerve sheath.

5. 희소돌기신경교세포를 표적하는 합성수용체를 포함하는 제1 제제; 및 상기 합성 수용체를 표적하는 합성 리간드를 포함하는 제2 제제를 포함하는 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물.5. A first agent containing a synthetic receptor targeting oligodendrocytes; and a second agent comprising a synthetic ligand targeting the synthetic receptor. A composition for improving memory, exercise and cognitive abilities.

6. 위 5에 있어서, 상기 합성수용체는 Gq-DREADD, Gs-DREADD 또는 β-arrestin preferring DREADD로 이루어진 군으로부터 선택되는 하나 이상인, 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물.6. The composition according to 5 above, wherein the synthetic receptor is at least one selected from the group consisting of Gq-DREADD, Gs-DREADD, or β-arrestin preferring DREADD, enhancing memory, improving motor skills, and improving cognitive abilities.

7. 위 5에 있어서, 상기 합성 리간드는 클로자핀 N-옥사이드, 클로자핀 및 컴파운드-21로 이루어진 군으로부터 선택되는 하나 이상인, 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물.7. The composition according to 5 above, wherein the synthetic ligand is one or more selected from the group consisting of clozapine N-oxide, clozapine, and compound-21, enhancing memory, improving athletic ability, and improving cognitive ability.

8. 위 5에 있어서, 상기 제1 제제 및 제 2 제제는 동시에 또는 순차적으로 처리될 수 있는, 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물.8. The composition according to 5 above, wherein the first agent and the second agent can be processed simultaneously or sequentially, enhancing memory, improving motor ability, and improving cognitive ability.

본 발명은 신경수초 가소성 증진용 조성물을 제공한다.The present invention provides a composition for enhancing nerve myelin plasticity.

본 발명 조성물의 제 1 제제는 희소돌기신경교세포에서 합성 수용체가 발현되도록 하고 제 2제제는 합성 수용체를 표적하여 희소돌기신경교세포를 자극함으로써, 인산화된 ERK 1/2를 증진시킬 수 있고, 이를 통해 신경수초의 가소성을 증진시킬 수 있다.The first agent of the composition of the present invention causes the expression of a synthetic receptor in oligodendrocytes, and the second agent targets the synthetic receptor to stimulate oligodendrocytes, thereby enhancing phosphorylated ERK 1/2. It can enhance the plasticity of the nerve sheath.

본 발명은 기억력, 인지능력 및 운동능력 증진방법을 제공한다.The present invention provides a method for improving memory, cognitive ability and motor ability.

본 발명은 희소돌기신경교세포에 합성 수용체를 전달하고, 합성수용체를 자극하는 합성 리간드를 처리함으로써, 신경수초의 가소성을 증진시킬 수 있고, 이를 통해 기억력, 인지능력 및 운동능력을 증진시킬 수 있다.The present invention can enhance the plasticity of nerve sheath by delivering synthetic receptors to oligodendrocytes and treating them with synthetic ligands that stimulate the synthetic receptors, thereby enhancing memory, cognitive ability and motor skills.

도 1은 합성수용체가 발현된 희소돌기신경교세포에 합성 리간드를 처리하여 신경수초의 가소성을 증진시키는 개략도이다(01: 희소돌기신경교세포, 02: 합성리간드, 03: ERK1/2 인산화, 04: 신경수초).
도 2는 Oligodendrocyte 특이적 hM3Dq유전자 발현 및 자극을 위한 CNO 전달 실험의 타임라인을 나타낸다.
도 3은 직팽창기법 및 면역조직학을 통한 oligodendrocyte의 인산화된 ERK1/2의 반응을 나타낸다.
도 4는 조직팽창기법 및 면역조직학을 통한 oligodendrocyte의 paranode의 길이 변화를 나타낸다.
도 5는 Oligodendrocyte 자극에 의한 myelin paranode (Caspr)의 길이 증가를 정량화한 것이다.1 is a schematic diagram of enhancing the plasticity of neural sheath by treating oligodendrocytes expressing synthetic receptors with synthetic ligands (01: oligodendrocytes, 02: synthetic ligands, 03: ERK1/2 phosphorylation, 04: nerve water plant).
Figure 2 shows the timeline of CNO delivery experiments for oligodendrocyte-specific hM3Dq gene expression and stimulation.
Figure 3 shows the response of phosphorylated ERK1/2 in oligodendrocytes through direct expansion technique and immunohistology.
Figure 4 shows the length change of the paranode of oligodendrocyte through tissue expansion technique and immunohistology.
Figure 5 quantifies the increase in the length of myelin paranode (Caspr) by oligodendrocyte stimulation.

본 발명은 희소돌기신경교세포를 자극하여 희소돌기신경교세포 내 ERK1/2를 인산화시킴으로써 신경수초의 가소성을 증진시킬 수 있는 방법에 관한 것이다. 또한, 본 발명은 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물에 관한 것이다.The present invention relates to a method capable of enhancing the plasticity of nerve sheath by stimulating oligodendrocytes to phosphorylate ERK1/2 in oligodendrocytes. In addition, the present invention relates to a composition for improving memory, motor ability and cognitive ability.

용어 "ERK 1/2"는 세포 외 신호 조절 키나아제(Extracellular signal-regulated protein kinases) 1 및 2를 통칭한다.The term "ERK 1/2" refers collectively to Extracellular signal-regulated protein kinases 1 and 2.

용어 "pERK 1/2"는 인산화 된 세포 외 신호 조절 키나아제(phosphorylated Extracellular signal-regulated protein kinases) 1 및 2를 통칭한다, The term "pERK 1/2" refers collectively to phosphorylated extracellular signal-regulated protein kinases 1 and 2.

신경수초 가소성(myelin plasticity)은 신경 수초의 구조, 밀도 및 기능의 변화를 포괄하는 것으로, 신경수초의 가소성을 증진시키는 것은 예컨대 신경수초의 길이, 두께, 밀도 및 분포 밀도를 증진시키는 것일 수 있다.Myelin plasticity encompasses changes in the structure, density, and function of the nerve sheath, and enhancing the plasticity of the nerve sheath may include, for example, enhancing the length, thickness, density, and distribution density of the nerve sheath.

본 발명은 희소돌기신경교세포에 합성수용체를 발현시키는 단계; 상기 합성 수용체를 표적하는 합성 리간드를 처리하여 ERK1/2를 인산화시키는 단계를 포함하는 신경 수초의 가소성 증진 방법을 제공한다.The present invention comprises the steps of expressing a synthetic receptor in oligodendrocytes; Provided is a method for promoting plasticity of nerve sheath, comprising the step of phosphorylating ERK1/2 by treating a synthetic ligand targeting the synthetic receptor.

본 발명 신경수초의 가소성을 증진시킬 수 있는 방법은 인간을 제외한 동물에 적용하는 것일 수 있다.The method capable of enhancing the plasticity of the nerve sheath of the present invention may be applied to animals other than humans.

본 발명에서 합성 수용체는 특정 물질에 의해 배타적으로 활성화되는 수용체로, Designer Receptors Exclusively Activated by Designer Drugs(DREADD)와 상호교환적으로 사용 가능하다.In the present invention, the synthetic receptor is a receptor that is exclusively activated by a specific substance, and can be used interchangeably with Designer Receptors Exclusively Activated by Designer Drugs (DREADD).

합성수용체는 예컨대 Gq-DREADD, Gs-DREADD 또는 β-arrestin preferring DREADD 등일 수 있다.The synthetic receptor may be, for example, Gq-DREADD, Gs-DREADD or β-arrestin preferring DREADD.

Gq-DREADD의 일종인 hM3Dq, hM1Dq 및 hM5Dq는 인간 무스카린성 수용체를 변형시킨 것으로, 클로자핀-N-옥사이드(CNO) 등의 합성 리간드에 의해 활성화되고 Gq 신호 전달 경로에 관여하며, 세포를 자극할 수 있다. hM3Dq, hM1Dq and hM5Dq, which are types of Gq-DREADD, are modified human muscarinic receptors, activated by synthetic ligands such as clozapine-N-oxide (CNO), involved in the Gq signaling pathway, and capable of stimulating cells. can

희소돌기신경교세포에 합성 수용체를 발현시키는 방법은 합성 수용체를 코딩하는 유전자를 포함하는 재조합 벡터를 희소돌기신경교세포에 투여하는 것일 수 있다. 예컨대 서열번호 1의 아미노산 서열로 이루어진 합성수용체 hM3Dq를 코딩하는 유전자를 포함하는 재조합 벡터일 수 있다. 서열번호 1의 아미노산 서열로 이루어진 합성수용체 hM3Dq를 코딩하는 유전자는 프로모터에 작동 가능하게 연결된다.A method of expressing a synthetic receptor in oligodendrocytes may be to administer a recombinant vector containing a gene encoding the synthetic receptor to oligodendrocytes. For example, it may be a recombinant vector containing a gene encoding the synthetic receptor hM3Dq consisting of the amino acid sequence of SEQ ID NO: 1. A gene encoding the synthetic receptor hM3Dq consisting of the amino acid sequence of SEQ ID NO: 1 is operably linked to a promoter.

용어 "작동 가능하게 연결(operably linked)"이란, 일반적 기능을 수행하도록 핵산 발현조절 서열과 목적하는 단백질 또는 RNA를 코딩하는 핵산 서열 이 기능적으로 연결(functional linkage)되어 있는 상태를 의미한다. 예를 들어 프로모터와 단백질 또는 RNA를 코딩하는 핵산 서열이 작동 가능하게 연결되어 코딩서열의 발현에 영향을 미칠 수 있다. 발현 벡터와의 작동 가능한 연결은 당해 기술분야에서 잘 알려진 유전자 재조합 기술을 이용하여 제조할 수 있으며, 부위-특이적 DNA 절단 및 연결은 당해 기술 분야에서 일반적으로 알려진 효소 등을 사용할 수 있다.The term "operably linked" refers to a state in which a nucleic acid expression control sequence and a nucleic acid sequence encoding a protein or RNA of interest are functionally linked to perform a general function. For example, a promoter and a nucleic acid sequence encoding a protein or RNA may be operably linked to affect expression of the coding sequence. Operable linkage with the expression vector can be prepared using gene recombination techniques well known in the art, and site-specific DNA cleavage and linkage can be performed using enzymes generally known in the art.

합성수용체 hM3Dq를 코딩하는 유전자는 예컨대 서열번호 2의 염기서열로 이루어진 것일 수 있다.The gene encoding the synthetic receptor hM3Dq may be composed of, for example, the nucleotide sequence of SEQ ID NO: 2.

합성수용체 hM3Dq를 코딩하는 유전자가 작동가능하게 연결된 프로모터는 PLP1(Proteolipid Protein 1) 프로모터(promoter)일 수 있다.A promoter to which a gene encoding the synthetic receptor hM3Dq is operably linked may be a Proteolipid Protein 1 (PLP1) promoter.

합성수용체 hM3Dq를 코딩하는 유전자가 PLP1 프로모터에 작동가능하게 연결된 재조합 벡터를 희소돌기신경교세포에 투여함으로써, 희도돌기신경교세포에서 hM3Dq를 특이적으로 발현시킬 수 있다.By administering to oligodendrocytes a recombinant vector in which the gene encoding the synthetic receptor hM3Dq is operably linked to the PLP1 promoter, hM3Dq can be specifically expressed in oligodendrocytes.

합성수용체를 코딩하는 유전자가 포함된 재조합 벡터의 제조방법은 제한되지 않으며, 예컨대 등록특허 제10-1964875에 기재된 방법으로 수행될 수 있다.A method for preparing a recombinant vector containing a gene encoding a synthetic receptor is not limited, and may be performed, for example, by the method described in Patent Registration No. 10-1964875.

제 1제제는 합성수용체 발현용 물질을 전달할 수 있는 바이러스를 더 포함할 수 있다. 합성수용체 발현용 물질을 전달할 수 있는 바이러스는 예컨대 아데노 부속 바이러스, 아데노 바이러스, 또는 렌티 바이러스일 수 있다.The first agent may further include a virus capable of delivering a material for expressing a synthetic receptor. A virus capable of delivering a material for expressing a synthetic receptor may be, for example, adeno-associated virus, adenovirus, or lentivirus.

희소돌기신경교세포에 합성 수용체를 발현시키는 방법은 합성 수용체를 코딩하는 유전자를 포함하는 재조합 벡터를 희소돌기신경교세포에 투여하는 것일 수 있다.A method of expressing a synthetic receptor in oligodendrocytes may be to administer a recombinant vector containing a gene encoding the synthetic receptor to oligodendrocytes.

합성 수용체, 합성 리간드 및 합성 수용체를 코딩하는 유전자를 포함하는 재조합 벡터는 전술한 바와 같다.A recombinant vector containing a synthetic receptor, a synthetic ligand, and a gene encoding the synthetic receptor is as described above.

희소돌기신경교세포에 합성수용체를 발현시키는 방법은 교배를 이용하는 것일 수 있다. 잠재적으로 합성 수용체를 발현할 수 있는 동물모델과 희소돌기신경교세포를 표적하는 동물모델을 교배시켜, 희소돌기신경교세포에서 합성수용체가 발현될 수 있는 개체를 얻을 수 있고, 상기 개체에 Tamoxifen을 투여함으로써 희소돌기신경교세포에서 hM3Dq가 발현되도록 할 수 있다.A method of expressing a synthetic receptor in oligodendrocytes may be to use mating. By crossing an animal model that can potentially express a synthetic receptor with an animal model targeting oligodendrocytes, an individual capable of expressing a synthetic receptor in oligodendrocytes can be obtained, and Tamoxifen is administered to the individual. hM3Dq can be expressed in oligodendrocytes.

본 발명의 일 실시예에서, hM3Dq의 유전자를 선택적으로 발현 가능한 개체(flex-hM3Dq; Jackson Laboratory, 026220) 와 희소돌기신경교세포에서 transgene(hM3Dq 유전자)의 발현을 유도하기위한 개체(PLP-CreERT)를 교배시켜 희소돌기신경교세포에 hM3Dq 합성수용체가 발현될 수 있는 마우스(PLP-hM3Dq transgenic mouse)를 제작하였다.In one embodiment of the present invention, an object capable of selectively expressing the gene of hM3Dq (flex-hM3Dq; Jackson Laboratory, 026220) and an object for inducing the expression of a transgene (hM3Dq gene) in oligodendrocytes (PLP-CreERT) was mated to prepare a mouse (PLP-hM3Dq transgenic mouse) capable of expressing the hM3Dq synthetic receptor in oligodendrocytes.

합성 수용체를 표적하는 합성 리간드는 상기 Gq-DREADD, Gs-DREADD 또는 β-arrestin preferring DREADD 등의 합성 수용체를 활성화시킬 수 있다. 합성 리간드는 예컨대 클로자핀 N-옥사이드(clozapine N-oxide, CNO), 클로자핀(clozapine, CLZ), 페르라핀(Perlapine) 또는 컴파운드-21(Compound-21, Cmpd-21) 등일 수 있다.Synthetic ligands targeting synthetic receptors can activate synthetic receptors such as Gq-DREADD, Gs-DREADD or β-arrestin preferring DREADD. The synthetic ligand may be, for example, clozapine N-oxide (CNO), clozapine (CLZ), perlapine or Compound-21 (Cmpd-21).

본 발명의 일 실시예에서, 마우스의 희소돌기신경교세포에 합성 수용체 hM3Dq를 발현시킨 후, 이를 활성화시키는 합성 리간드로 클로자핀 N-옥사이드(Clozapine N-oxide, CNO)를 처리함으로써, 신경수초의 가소성을 증진시켰다.In one embodiment of the present invention, after expressing the synthetic receptor hM3Dq in oligodendrocytes of a mouse, and then treating it with synthetic ligand activating clozapine N-oxide (Clozapine N-oxide, CNO), neuromyelin plasticity is improved. promoted

합성 리간드의 처리 방법은 대상에 경구 투여, 혈관 주사 또는 국소 주사하는 것일 수 있다. A method of administering the synthetic ligand may be oral administration, intravascular injection, or local injection to a subject.

희소돌기신경교세포에 합성 수용체가 발현된 대상에 합성 리간드를 처리함으로써, 희소돌기신경교세포 내 인산화된 ERK1/2가 증가되며, 이로 인해 신경수초의 가소성이 증진될 수 있다. 예컨대 신경수초의 길이, 두께, 밀도가 증가될 수 있고, 분포 밀도가 증가될 수 있다.By treating oligodendrocytes with synthetic ligands expressed in oligodendrocytes with synthetic ligands, phosphorylated ERK1/2 in oligodendrocytes is increased, thereby enhancing neuromyelin plasticity. For example, the length, thickness, and density of nerve sheaths may be increased, and distribution density may be increased.

또한, 본 발명은 희소돌기신경교세포를 표적하는 합성수용체 발현용 물질을 포함하는 제1 제제; 및 상기 합성 수용체를 표적하는 합성 리간드를 포함하는 제2 제제를 포함하는 신경수초 가소성 증진용 조성물을 제공한다.In addition, the present invention provides a first agent comprising a substance for expressing a synthetic receptor targeting oligodendrocytes; and a second agent comprising a synthetic ligand targeting the synthetic receptor.

희소돌기신경교세포를 표적하는 합성수용체 발현용 물질은 합성 수용체를 코딩하는 유전자를 포함하는 재조합 벡터일 수 있다. 예컨대 합성수용체 hM3Dq를 코딩하는 유전자(서열번호 1)를 포함하는 재조합 벡터일 수 있다.A material for expressing a synthetic receptor targeting oligodendrocytes may be a recombinant vector containing a gene encoding the synthetic receptor. For example, it may be a recombinant vector containing a gene encoding the synthetic receptor hM3Dq (SEQ ID NO: 1).

합성수용체는 예컨대 Gq-DREADD, Gs-DREADD 또는 β-arrestin preferring DREADD 등일 수 있다.The synthetic receptor may be, for example, Gq-DREADD, Gs-DREADD or β-arrestin preferring DREADD.

Gq-DREADD의 일종인 hM3Dq, hM1Dq 및 hM5Dq는 인간 무스카린성 수용체를 변형시킨 것으로, 클로자핀-N-옥사이드(CNO) 등의 합성 리간드에 의해 활성화되고 Gq 신호 전달 경로에 관여하며, 세포를 자극할 수 있다. hM3Dq, hM1Dq and hM5Dq, which are types of Gq-DREADD, are modified human muscarinic receptors, activated by synthetic ligands such as clozapine-N-oxide (CNO), involved in the Gq signaling pathway, and capable of stimulating cells. can

합성 수용체를 코딩하는 유전자가 포함된 재조합 벡터는 PLP1(Proteolipid Protein 1) 프로모터(promoter)를 더 포함할 수 있다.The recombinant vector containing the gene encoding the synthetic receptor may further include a Proteolipid Protein 1 (PLP1) promoter.

PLP1 프로모터를 더 포함함으로써, 희소돌기신경교세포에서 hM3Dq를 특이적으로 발현시킬 수 있다.By further including the PLP1 promoter, hM3Dq can be specifically expressed in oligodendrocytes.

합성 수용체를 코딩하는 유전자가 포함된 재조합 벡터의 제조는 등록특허 제10-1964875에 기재된 방법으로 수행될 수 있다.Preparation of a recombinant vector containing a gene encoding a synthetic receptor may be performed by the method described in Korean Patent Registration No. 10-1964875.

제 1제제는 합성수용체 발현용 물질을 전달할 수 있는 바이러스를 더 포함할 수 있다. 합성수용체 발현용 물질을 전달할 수 있는 바이러스는 예컨대 아데노 부속 바이러스, 아데노 바이러스, 또는 렌티 바이러스일 수 있다.The first agent may further include a virus capable of delivering a material for expressing a synthetic receptor. A virus capable of delivering a material for expressing a synthetic receptor may be, for example, adeno-associated virus, adenovirus, or lentivirus.

합성 수용체를 표적하는 합성 리간드는 상기 Gq-DREADD, Gs-DREADD 또는 β-arrestin preferring DREADD 등의 합성 수용체를 활성화시킬 수 있다. 합성 리간드는 예컨대 클로자핀 N-옥사이드(clozapine N-oxide, CNO), 클로자핀(clozapine, CLZ), 페르라핀(Perlapine) 또는 컴파운드-21(Compound-21, Cmpd-21) 등일 수 있다.Synthetic ligands targeting synthetic receptors can activate synthetic receptors such as Gq-DREADD, Gs-DREADD or β-arrestin preferring DREADD. The synthetic ligand may be, for example, clozapine N-oxide (CNO), clozapine (CLZ), perlapine or Compound-21 (Cmpd-21).

본 발명의 조성물은 신경수초의 가소성을 증진시킬 수 있다. 예컨대 신경수초의 길이, 두께, 밀도 및 분포 밀도를 증가시킬 수 있다.The composition of the present invention can enhance the plasticity of the nerve sheath. For example, the length, thickness, density and distribution density of nerve sheaths can be increased.

본 발명의 조성물은 신경수초의 길이, 두께, 밀도 및 분포 밀도를 증가시킴으로써 대상의 기억력 증진에 도움이 된다.The composition of the present invention helps improve the memory of a subject by increasing the length, thickness, density and distribution density of the nerve sheath.

본 발명의 조성물은 신경수초의 길이, 두께, 밀도 및 분포 밀도를 증가시킴으로써 인지능력 및 운동능력 증진에 도움이 된다.The composition of the present invention is helpful in improving cognitive ability and motor ability by increasing the length, thickness, density and distribution density of the nerve sheath.

본 발명의 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물은 건강기능식품으로 활용할 수 있다.The composition for improving memory, exercise and cognitive abilities of the present invention can be used as a health functional food.

본 발명의 건강기능식품이라 함은, 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품일 수 있고, 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.The health functional food of the present invention may be a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the Health Functional Food Act No. It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating or physiological functions.

본 발명의 건강기능식품은 통상의 식품 첨가물을 포함할 수 있으며, 식품 첨가물로서의 적합 여부는 다른 규정이 없는 한, 식품의약품안전청에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.The health functional food of the present invention may contain ordinary food additives, and the suitability as a food additive is determined according to the general rules of the Food Additive Code and General Test Methods approved by the Food and Drug Administration, unless otherwise specified. It is judged according to standards and standards.

상기 식품 첨가물 공전에 수재된 품목으로는 예를 들어, 케톤류, 글리신, 구연산칼슘, 니코틴산, 계피산 등의 화학적 합성물; 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물; L-글루타민산나트륨 제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류 등을 포함하나, 이에 제한되지 않는다.Items listed in the Food Additives Codex include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, kaoliang pigment, and guar gum; It includes, but is not limited to, mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.

정제 형태의 건강기능식품은 본 발명의 추출물을 부형제, 결합제, 붕해제 및 다른 첨가제와 혼합한 혼합물을 통상의 방법으로 과립화한 다음, 활택제 등을 넣어 압축성형하거나, 상기 혼합물을 직접 압축 성형할 수 있다. 또한 상기 정제 형태의 식품은 필요에 따라 교미제 등을 함유할 수도 있다.The health functional food in the form of a tablet is obtained by granulating a mixture obtained by mixing the extract of the present invention with excipients, binders, disintegrants, and other additives in a conventional manner, and then adding a lubricant or the like to compression molding or directly compressing the mixture. can do. In addition, the food in tablet form may contain a flavoring agent or the like as needed.

캅셀 형태의 건강기능식품 중 경질 캅셀제는 통상의 경질 캅셀에 본 발명의 추출물을 부형제 등의 첨가제와 혼합한 혼합물을 충진하여 제조할 수 있으며, 연질 캅셀제는 본 발명의 추출물을 부형제 등의 첨가제와 혼합한 혼합물을 젤라틴과 같은 캅셀기제에 충진하여 제조할 수 있다. 상기 연질 캅셀제는 필요에 따라 글리세린 또는 소르비톨 등의 가소제, 착색제, 보존제 등을 함유할 수 있다.Among health functional foods in the form of capsules, hard capsules can be prepared by filling a mixture of the extract of the present invention mixed with additives such as excipients in a normal hard capsule, and soft capsules can be prepared by mixing the extract of the present invention with additives such as excipients. It can be prepared by filling a mixture into a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.

환 형태의 건강기능식품은 본 발명의 추출물과 부형제, 결합제, 붕해제 등을 혼합한 혼합물을 기존에 공지된 방법으로 성형하여 조제할 수 있으며, 필요에 따라 백당이나 다른 제피제로 제피할 수 있으며, 또는 전분, 탈크와 같은 물질로 표면을 코팅할 수도 있다.The health functional food in the form of a pill can be prepared by molding a mixture of the extract of the present invention mixed with an excipient, a binder, a disintegrant, etc. by a conventionally known method, and can be coated with sucrose or other coating agents if necessary, Alternatively, the surface may be coated with a material such as starch or talc.

과립 형태의 건강기능식품은 본 발명의 추출물과 부형제, 결합제, 붕해제 등을 혼합한 혼합물을 기존에 공지된 방법으로 입상으로 제조할 수 있으며, 필요에 따라 착향제, 교미제 등을 함유할 수 있다.A health functional food in the form of a granule may be prepared in a granular form by a previously known method of a mixture of the extract of the present invention mixed with an excipient, a binder, a disintegrant, etc., and may contain flavoring agents, flavoring agents, etc., if necessary. there is.

이하, 본 발명을 실시예로 보다 상세하게 설명한다.Hereinafter, the present invention will be described in more detail with examples.

실시예Example

1. 희소돌기신경교세포에 hM3Dq 합성수용체를 발현시킨 마우스의 제작1. Production of mice expressing hM3Dq synthetic receptor in oligodendrocytes

hM3Dq의 유전자를 선택적으로 발현 가능한 개체와(flex-hM3Dq; Jackson Laboratory, 026220) 희소돌기신경교세포에만 유전자를 국소화 하기 위한 개체(PLP-CreERT; Jackson Laboratory,005975)를 교배하여 희소돌기신경교세포에 hM3Dq 합성수용체를 발현시킨 마우스(PLP-hM3Dq transgenic mouse)를 제작하였다.hM3Dq in oligodendrocytes by crossing an individual capable of selectively expressing the hM3Dq gene (flex-hM3Dq; Jackson Laboratory, 026220) and an individual capable of localizing the gene only to oligodendrocytes (PLP-CreERT; Jackson Laboratory, 005975). A mouse expressing the synthetic receptor (PLP-hM3Dq transgenic mouse) was prepared.

2. 합성 리간드의 처리2. Treatment of synthetic ligands

희소돌기신경교세포에 hM3Dq 합성수용체를 발현시킨 마우스(PLP-hM3Dq transgenic mouse)의 생후 80일경 tamoxifen을 해바라기씨유(sigma, S5007)에 10 mg/mL의 농도로 녹여 100 μL만큼 7일 연속으로 복강주사 하였다. 그 다음 7일간의 휴식기간 후 17일동안 CNO를 생리식염수에 용해한 용액을 마우스에 복강 주사하였으며, 매일 1 mg/kg(CNO 양/실험동물의 무게)의 비율로 주사하였다. 마지막 복강 주사 후 10일간의 휴식기를 주어 유전자 발현이 충분히 되도록 유도했다(도 2).Around 80 days after birth of a mouse (PLP-hM3Dq transgenic mouse) in which the hM3Dq synthetic receptor was expressed in oligodendrocytes, tamoxifen was dissolved in sunflower seed oil (sigma, S5007) at a concentration of 10 mg/mL and administered intraperitoneally as 100 μL for 7 consecutive days. Injected. Then, for 17 days after a rest period of 7 days, mice were intraperitoneally injected with a solution in which CNO was dissolved in physiological saline, and injected at a daily rate of 1 mg/kg (CNO amount/weight of experimental animal). After the last intraperitoneal injection, a 10-day rest period was given to induce sufficient gene expression (FIG. 2).

3. 조직처리3. Tissue processing

합성 리간드(CNO)를 처리하고 10일간의 휴식기 이후 해상도 및 해상도 및 세포/단백질 발현을 확인하기 위한 조직처리(조직팽창기법, 면역조직학)를 진행하였다.After treatment with synthetic ligand (CNO) and a rest period of 10 days, tissue processing (tissue expansion technique, immunohistology) was performed to confirm resolution and resolution and cell/protein expression.

실험동물을 마취한 후 자극 반응이 없는 것을 확인한 뒤, 생리식염수를 이용해 체내 혈액을 제거하고 4% 파라 포름알데히드로 조직 고정을 수행하였다. 적출된 뇌 조직은 4% 파라포름알데히드, 4% 아크릴아마이드가 혼합된 PBS용액에 12시간동안 배양하였으며, 배양 후 vibratome (VT1200s, leica)를 통하여 130 um의 두께로 균일하게 잘라냈다.After confirming that there is no stimulation response after anesthetizing the experimental animal, body blood was removed using physiological saline, and tissue fixation was performed with 4% paraformaldehyde. The extracted brain tissue was cultured for 12 hours in a PBS solution containing 4% paraformaldehyde and 4% acrylamide, and then uniformly cut to a thickness of 130 um through a vibratome (VT1200s, leica).

잘라낸 뇌 조직을 19% sodium acrylate, 10% acrylamide, 0.1% bis acrylate, VA-044 (20mg)을 섞은 혼합용액에 24시간동안 배양했으며, 40 ℃ 인큐베이터에서 gelation을 통하여 tissue-gel 혼합체를 만들었다. 만들어진 tissue-gel 혼합체는 Sodium dodecyl sulfate 용액을 이용해 단백질이 분해되어 ‘clearing’ 되도록 했다. 이후 3일간 primary antibody에 배양하여 표적 단백질의 면역염색기법을 통해 지표 하고 난 뒤, 1일간 secondary antibody에 배양하여 형광지표자를 부착했다. 영상분석 수행 3시간전에 cleared tissue-gel 면역염색 시료는 3차 증류수에서 수분 흡수를 통해 본래 크기의 약 4배 (부피 64배)의 팽창된 시료가 사용되었다.The cut brain tissue was cultured in a mixture of 19% sodium acrylate, 10% acrylamide, 0.1% bis acrylate, and VA-044 (20 mg) for 24 hours, and a tissue-gel mixture was prepared through gelation in an incubator at 40 °C. The resulting tissue-gel mixture was 'cleared' by protein degradation using a sodium dodecyl sulfate solution. After that, it was cultured in the primary antibody for 3 days to indicate the target protein through immunostaining technique, and then cultured in the secondary antibody for 1 day to attach a fluorescent indicator. 3 hours before image analysis, a sample expanded to about 4 times the original size (64 times the volume) through water absorption in tertiary distilled water was used as the cleared tissue-gel immunostaining sample.

4. ERK1/2단백질 발현의 증가 확인4. Confirmation of increase in ERK1/2 protein expression

Tamoxifen에 의해 희소돌기신경교세포에 합성리간드 수용체가 발현된 동물들을 실험군과 대조군으로 나누었다. 실험군은 1mg/kg 농도의 합성리간드를 복강주사를 통해 연속된 17일동안 주사하였으며, 대조군은 17일동안 생리식염수를 동일 부피로 복강주사 하였다.Animals in which synthetic ligand receptors were expressed in oligodendrocytes by Tamoxifen were divided into experimental and control groups. The experimental group was injected intraperitoneally with the synthetic ligand at a concentration of 1 mg/kg for 17 consecutive days, and the control group was intraperitoneally injected with the same volume of physiological saline for 17 days.

주사 17일 이후, 조직 팽창 현미경 및 면역염색실험법을 통해 실험동물의 뇌 조직을 관찰했다.After 17 days of injection, the brain tissues of the experimental animals were observed using a tissue expansion microscope and an immunostaining method.

oligodendrocyte의 myelin 구조적 변화가 유도되기 위한 지표로, CNPase(biolegend, #836404)항체를 통한 oligodendrocyte의 표지와 인산화된 ERK1/2의 세포 내 발현을 확인하기 위한 pERK1/2 항체(cell signaling technology, #4370s)를 통해 비교해 본 결과, 대조군(Sham control)보다 높은 비율로 oligodendrocyte내 인산화된 ERK1/2단백질의 발현이 증가함을 확인하였다(도 3).As an indicator for the induction of myelin structural changes in oligodendrocytes, oligodendrocyte labeling through CNPase (biolegend, #836404) antibody and pERK1/2 antibody (cell signaling technology, #4370s) to confirm intracellular expression of phosphorylated ERK1/2 As a result of comparison through ), it was confirmed that the expression of phosphorylated ERK1/2 protein in oligodendrocyte increased at a higher rate than the control group (Sham control) (FIG. 3).

성숙한 희소돌기신경교세포를 표지하는 CNPase(적색)과 인산화된 ERK1/2단백질(녹색)의 염색을 통해 전체 희소돌기신경교세포 중, 인산화된 ERK1/2단백질을 발현하는 희소돌기신경교세포를 비교했다(도 3). 도 3의 Sham control(좌)의 경우 약 10% 정도의 희소돌기신경교세포가 인산화된 ERK1/2를 발현한 반면에, 합성리간드(CNO)를 처리한 CNO administration(우)의 경우, 약 90% 정도의 희소돌기신경교세포가 인산화된 ERK1/2를 발현하였다.Through staining of CNPase (red) and phosphorylated ERK1/2 protein (green), which mark mature oligodendrocytes, oligodendrocytes expressing phosphorylated ERK1/2 protein were compared among all oligodendrocytes ( Fig. 3). In the case of Sham control (left) in FIG. 3, about 10% of oligodendrocytes expressed phosphorylated ERK1/2, whereas in the case of CNO administration treated with synthetic ligand (CNO), about 90% A few oligodendrocytes expressed phosphorylated ERK1/2.

5. 희소돌기신경교세포 신경수초 파라노드의 길이 변화5. Changes in the length of oligodendrocyte paranodes in neuromyelin cells

성숙한 oligodendrocyte의 구조체인 myelin의 말단부(파라노드)의 길이변화를 측정하기 위해, paranode를 표지하는 Caspr(abcam, ab133634) 항체를 통한 파라노드를 표지하였음. Sham control에 비해 Caspr의 길이가 평균적으로 CNO를 투여한 마우스에서 더 길어짐을 관측하였다(도 4).In order to measure the length change of the distal end (paranode) of myelin, a structure of a mature oligodendrocyte, the paranode was labeled with Caspr (abcam, ab133634) antibody that labels the paranode. Compared to the sham control, it was observed that the average length of Caspr was longer in mice administered with CNO (FIG. 4).

6. 희소돌기신경교세포 자극에 의한 신경수초 파라노드의 길이 증가 정량화6. Quantification of the increase in the length of the neuromyelin paranode by oligodendrocyte stimulation

합성 리간드(CNO)를 처리하였을 때, hM3Dq를 통해 희소돌기신경교세포 신경수초의 파라노드(Caspr)의 길이가 평균적으로 약 0.7um 증가하였다(도 5). 또한, 파라노드의 분포 또한 증가함을 확인할 수 있었다 (박스 사이즈: 25~75 percentile. 박스 내부 적색선: 평균 paranode 길이. 하단 및 상단 수염: 데이터 최소/최대 값).When the synthetic ligand (CNO) was treated, the length of the paranode (Caspr) of the oligodendrocyte nerve sheath was increased by about 0.7 μm on average through hM3Dq (FIG. 5). In addition, it was confirmed that the distribution of paranodes also increased (box size: 25 to 75 percentile. Red line inside the box: average paranode length. Lower and upper whiskers: data minimum/maximum values).

<110> SEOUL NATIONAL UNIVERSITY R&DB FOUNDATION <120> Method for promoting myelin plasticity <130> 21P10002 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 590 <212> PRT <213> Artificial Sequence <220> <223> hM3Dq <400> 1 Met Thr Leu His Asn Asn Ser Thr Thr Ser Pro Leu Phe Pro Asn Ile 1 5 10 15 Ser Ser Ser Trp Ile His Ser Pro Ser Asp Ala Gly Leu Pro Pro Gly 20 25 30 Thr Val Thr His Phe Gly Ser Tyr Asn Val Ser Arg Ala Ala Gly Asn 35 40 45 Phe Ser Ser Pro Asp Gly Thr Thr Asp Asp Pro Leu Gly Gly His Thr 50 55 60 Val Trp Gln Val Val Phe Ile Ala Phe Leu Thr Gly Ile Leu Ala Leu 65 70 75 80 Val Thr Ile Ile Gly Asn Ile Leu Val Ile Val Ser Phe Lys Val Asn 85 90 95 Lys Gln Leu Lys Thr Val Asn Asn Tyr Phe Leu Leu Ser Leu Ala Cys 100 105 110 Ala Asp Leu Ile Ile Gly Val Ile Ser Met Asn Leu Phe Thr Thr Tyr 115 120 125 Ile Ile Met Asn Arg Trp Ala Leu Gly Asn Leu Ala Cys Asp Leu Trp 130 135 140 Leu Ala Ile Asp Cys Val Ala Ser Asn Ala Ser Val Met Asn Leu Leu 145 150 155 160 Val Ile Ser Phe Asp Arg Tyr Phe Ser Ile Thr Arg Pro Leu Thr Tyr 165 170 175 Arg Ala Lys Arg Thr Thr Lys Arg Ala Gly Val Met Ile Gly Leu Ala 180 185 190 Trp Val Ile Ser Phe Val Leu Trp Ala Pro Ala Ile Leu Phe Trp Gln 195 200 205 Tyr Phe Val Gly Lys Arg Thr Val Pro Pro Gly Glu Cys Phe Ile Gln 210 215 220 Phe Leu Ser Glu Pro Thr Ile Thr Phe Gly Thr Ala Ile Ala Gly Phe 225 230 235 240 Tyr Met Pro Val Thr Ile Met Thr Ile Leu Tyr Trp Arg Ile Tyr Lys 245 250 255 Glu Thr Glu Lys Arg Thr Lys Glu Leu Ala Gly Leu Gln Ala Ser Gly 260 265 270 Thr Glu Ala Glu Thr Glu Asn Phe Val His Pro Thr Gly Ser Ser Arg 275 280 285 Ser Cys Ser Ser Tyr Glu Leu Gln Gln Gln Ser Met Lys Arg Ser Asn 290 295 300 Arg Arg Lys Tyr Gly Arg Cys His Phe Trp Phe Thr Thr Lys Ser Trp 305 310 315 320 Lys Pro Ser Ser Glu Gln Met Asp Gln Asp His Ser Ser Ser Asp Ser 325 330 335 Trp Asn Asn Asn Asp Ala Ala Ala Ser Leu Glu Asn Ser Ala Ser Ser 340 345 350 Asp Glu Glu Asp Ile Gly Ser Glu Thr Arg Ala Ile Tyr Ser Ile Val 355 360 365 Leu Lys Leu Pro Gly His Ser Thr Ile Leu Asn Ser Thr Lys Leu Pro 370 375 380 Ser Ser Asp Asn Leu Gln Val Pro Glu Glu Glu Leu Gly Met Val Asp 385 390 395 400 Leu Glu Arg Lys Ala Asp Lys Leu Gln Ala Gln Lys Ser Val Asp Asp 405 410 415 Gly Gly Ser Phe Pro Lys Ser Phe Ser Lys Leu Pro Ile Gln Leu Glu 420 425 430 Ser Ala Val Asp Thr Ala Lys Thr Ser Asp Val Asn Ser Ser Val Gly 435 440 445 Lys Ser Thr Ala Thr Leu Pro Leu Ser Phe Lys Glu Ala Thr Leu Ala 450 455 460 Lys Arg Phe Ala Leu Lys Thr Arg Ser Gln Ile Thr Lys Arg Lys Arg 465 470 475 480 Met Ser Leu Val Lys Glu Lys Lys Ala Ala Gln Thr Leu Ser Ala Ile 485 490 495 Leu Leu Ala Phe Ile Ile Thr Trp Thr Pro Tyr Asn Ile Met Val Leu 500 505 510 Val Asn Thr Phe Cys Asp Ser Cys Ile Pro Lys Thr Phe Trp Asn Leu 515 520 525 Gly Tyr Trp Leu Cys Tyr Ile Asn Ser Thr Val Asn Pro Val Cys Tyr 530 535 540 Ala Leu Cys Asn Lys Thr Phe Arg Thr Thr Phe Lys Met Leu Leu Leu 545 550 555 560 Cys Gln Cys Asp Lys Lys Lys Arg Arg Lys Gln Gln Tyr Gln Gln Arg 565 570 575 Gln Ser Val Ile Phe His Lys Arg Ala Pro Glu Gln Ala Leu 580 585 590 <210> 2 <211> 9121 <212> DNA <213> Artificial Sequence <220> <223> hM3Dq <400> 2 acaccagctc tctcagtccg gggaggagga ggagcaggag gaacgcgagg aggaaggaga 60 ggaggagcgg ccagcagtag ccacgaccgc caccaccagg cagaggaaga gttcgtgggg 120 aggaaaagac ctctcctccc ccttggagcg ccttctctct gcttttggag aaagggaata 180 cacgataaag aaggagacgg aaagaagaga aaaagtgagg cgggagacag agggaaaagg 240 gcgtgaacag aaagggccgg agcgtgcagg ggagcacagg gcgcgggggc ggcactgccg 300 agccgggagc gctgccgctt gggcaggtgc cgcggccgct gcccggaggc ggcatgtgac 360 gcgcggccgc agctgcccgc gggcggagcg ctctcagacc ccggagcgca caccgcgggg 420 ccatcggtgc catcgcggat ctccaggctc ctcatcagtc cgccggggcc gcagcagcgc 480 ttctgggaag acgggcgatg aactgaaggg cggctccggg caggggggca cgatcttaag 540 gacagtcgct ccctgaacgc ggagccggag gagacgaagg gaaggtggag cggacgccac 600 ccgcgcaccg ggcaggcgcg gagaccggcg tgggacagcc acctggagcg cagctgccag 660 atatgaacag taatggcaca tattttggtt atgagtcact cagaggactg tggattgaat 720 gaactgtatc catccccatc atgatgtaca gaaccaagtc tcttcactag ttttgctgtg 780 gcgtggcacc tggtctcttt ctagaaggaa agttcaacat acagcacaat tctggacaca 840 ttgtattggt ttgatgctcc tacctggaac agatcctcat gagctagtaa gtttgaaggg 900 gagtcataaa gaggaacagg cattatacgc aaggctgaag aagcagttgg taactaacag 960 tacatctttt tctaaggtgg agctggtctc ttgggcagcc tgacatctgg tccactcctc 1020 tgtcctatgc cgggatcatc atgaccgtag agattatgtc actgttttgc atccttgtta 1080 cataactcag ttcctggtag attactatgt cagagagtca caatgacctt gcacaataac 1140 agtacaacct cgcctttgtt tccaaacatc agctcctcct ggatacacag cccctccgat 1200 gcagggctgc ccccgggaac cgtcactcat ttcggcagct acaatgtttc tcgagcagct 1260 ggcaatttct cctctccaga cggtaccacc gatgaccctc tgggaggtca taccgtctgg 1320 caagtggtct tcatcgcttt cttaacgggc atcctggcct tggtgaccat catcggcaac 1380 atcctggtaa ttgtgtcatt taaggtcaac aagcagctga agacggtcaa caactacttc 1440 ctcttaagcc tggcctgtgc cgatctgatt atcggggtca tttcaatgaa tctgtttacg 1500 acctacatca tcatgaatcg atgggcctta gggaacttgg cctgtgacct ctggcttgcc 1560 attgactacg tagccagcaa tgcctctgtt atgaatcttc tggtcatcag ctttgacaga 1620 tacttttcca tcacgaggcc gctcacgtac cgagccaaac gaacaacaaa gagagccggt 1680 gtgatgatcg gtctggcttg ggtcatctcc tttgtccttt gggctcctgc catcttgttc 1740 tggcaatact ttgttggaaa gagaactgtg cctccgggag agtgcttcat tcagttcctc 1800 agtgagccca ccattacttt tggcacagcc atcgctgctt tttatatgcc tgtcaccatt 1860 atgactattt tatactggag gatctataag gaaactgaaa agcgtaccaa agagcttgct 1920 ggcctgcaag cctctgggac agaggcagag acagaaaact ttgtccaccc cacgggcagt 1980 tctcgaagct gcagcagtta cgaacttcaa cagcaaagca tgaaacgctc caacaggagg 2040 aagtatggcc gctgccactt ctggttcaca accaagagct ggaaacccag ctccgagcag 2100 atggaccaag accacagcag cagtgacagt tggaacaaca atgatgctgc tgcctccctg 2160 gagaactccg cctcctccga cgaggaggac attggctccg agacgagagc catctactcc 2220 atcgtgctca agcttccggg tcacagcacc atcctcaact ccaccaagtt accctcatcg 2280 gacaacctgc aggtgcctga ggaggagctg gggatggtgg acttggagag gaaagccgac 2340 aagctgcagg cccagaagag cgtggacgat ggaggcagtt ttccaaaaag cttctccaag 2400 cttcccatcc agctagagtc agccgtggac acagctaaga cttctgacgt caactcctca 2460 gtgggtaaga gcacggccac tctacctctg tccttcaagg aagccactct ggccaagagg 2520 tttgctctga agaccagaag tcagatcact aagcggaaaa ggatgtccct ggtcaaggag 2580 aagaaagcgg cccagaccct cagtgcgatc ttgcttgcct tcatcatcac ttggacccca 2640 tacaacatca tggttctggt gaacaccttt tgtgacagct gcatacccaa aaccttttgg 2700 aatctgggct actggctgtg ctacatcaac agcaccgtga accccgtgtg ctatgctctg 2760 tgcaacaaaa cattcagaac cactttcaag atgctgctgc tgtgccagtg tgacaaaaaa 2820 aagaggcgca agcagcagta ccagcagaga cagtcggtca tttttcacaa gcgcgcaccc 2880 gagcaggcct tgtagaatga ggttgtatca atagcagtga caaaacgcac acatcaaccc 2940 acagacctta ggaggaggaa ggcgagggcg gggtgacttc tggtgatgat aaaaatggtt 3000 ttatcaccca gatgtgaaag aagctgcctg tttactgatc cattgaataa acccatttta 3060 atagaaaaag tcaataccaa ttcagcaaaa agaaaaaaaa aacatactac tgaatataaa 3120 gaaatttatt ctgaaataga ctttacgtgt ttttttctta aagaggagaa aaatattgct 3180 tgacggcaat tatataccca aagtgatttg cctgggtcct ttaattccca ttagctttgg 3240 aatctcagat gagcatagct gacccagttc ccacattctt cccaaggatc ccaaaagtgg 3300 gaatccagac cccaagtgga acactgcagg cttacgaatc tgtggttcca aaattatttc 3360 atacgttgca aagctgaatc ttcttgtccc aatagagctt cctgtctttt ctttggtgtg 3420 ttgttaaact ctatttgtgg acttgattct tgattcttgc aaagtactgt tttgtgcagt 3480 tcaagtttcg tacaaataaa aatacttaag tatatatata tgtgtgagtt ctgcacgcac 3540 acacatagtg tatataatat catgggaaac actgaactgg caaattattc ctgcaacata 3600 cgctttcagt actttggtaa ctgaagttct ctaggatcct aatgcaacat taacgtgaaa 3660 taagcccagt gtaatgtttt tgaaaccagg gctgttttcc acagagagca gccaggcctt 3720 cccagcaggt ctgtgcagag cggacaggct cgtgagtcag ctgagcgccg tggcttcgcc 3780 agacttggtg ttaagcaacc tcctttgttg atgtctcaac agagctaaat cggggcccct 3840 ctgagctcaa agaatgaacc acatccacac gtttgaattt aatcatctaa atctgaatgt 3900 ttcagaacaa aatttctgct atctaaactg cttgaaactc aataatagtg tcacgtttga 3960 atgtcataca cagcaatata tatatatgtg tatatatata tatatggcaa agcaaaaaaa 4020 aaaacatggt aagagagaat gaaggagaac attgtgtttg attcttgctg aatggcacct 4080 tctcaaagaa aatagggctt gcacctttgt taatcagctg tggccagtgc tttctggtgt 4140 tcattgtgta accttcaccc aggaataggt gaggttttag gaagttacat gtcctctgaa 4200 gaaagaatta cactctgaaa agtaatgctt caaattgatt tccttacctt ttgggaaaaa 4260 aaaaaaattg tttttttgca ttctcccttg aattgaccaa aatgttaact gtttcatttg 4320 gggaggggat ggggtgctgc catcattgtc gttgttgttg ctgctgtagc tgttggggtt 4380 tcttttcctg ttgccggggc tgtttgggga gagggagggg agggaggtgg gagggccgcg 4440 gagatatctt ccctttgtac agggcattcg tgttgtgaac ccagagctgg gtagaagctg 4500 cttttgtatt cagtgtgagg tggtgtttac agacgacttt gacaacagta gaagtgtact 4560 cagtggtgtc tgtgtatctg aactatttaa tttcgtgtta tgtttatatg cagaaatatt 4620 tatggatact acaccaagtg tttatttaat gttgataaat atgactcttc agtcgtcagc 4680 catggtgtcc tttcaaagtg attctttaag gtccacttga gcaatgaata gagtatattg 4740 gagctttcct gtggctaaga agaagaaaca tgtcatcctg ttggcatcac caagcaccta 4800 actctttcta ggtaataaaa agtcaacggt cacttcaacg taaatgttta aaaccactct 4860 ttagttaaat ctgatgggaa ctgaacaaag ctccaccatt gtgtctgaaa tcattttaca 4920 cagggaaaaa aaaaaacaat cagcataatt gtaagaatga tttttttggg cccacattca 4980 accaaagaac aagaaagaaa cagacgattt tctaatatgt acacagcaaa ctacagaatg 5040 tcatattatc cacctgtgaa acatctagta agcaattgca tctatatttt atttctcaag 5100 ttggcctggc actaataaat atgtgaaagc ttattcaagt gtcccagtac ttcagcactt 5160 cccatccttt tggaaactcc tatcttgcct ccctcccaaa agggctgttt cttttattaa 5220 aaaaaaaaaa gaaatagaat taaatttgag aaaaggagaa accctgaaat ttaacagaac 5280 tttgagagtc acttccaatc tccttaaaca tgattatctc cttatccttg tccttttgtg 5340 atgctaataa tagcagaaaa ggaacaaatc caagactcta ggtctccctt ttttgtttta 5400 ctggttttat tatagaattg tttttaagcc tcttatggat gaaagctgca tttaaatact 5460 atctactgat tttatagaaa ggtatgtcca tttcttgggt cactttcaga gaaagaaaac 5520 aagcaaaata ggttcttttg aatatgaata atttaaaagc ataattgaag atgtagtgag 5580 tatttataat tggaccttag ttacacgtgc atttctcaaa gggcttgtca ttacaggaga 5640 tattcagcac ttacctaaga tcaaaatgtc ttcatcccaa attgggaatc gaattatttc 5700 atgccatatg catgaggttt gcagacattt caagattagc cactttgctt gaaagaagaa 5760 aatgatcaaa cctaaaataa actacacatg gagaatctct ctctctctct ctgcctctct 5820 ctctctcttt ctccctcttc ttccttctct ctctctcaca agcacacaca cacacactca 5880 atctccattc tccaggcatt gccagatggt taggagaagt gagctctctc tggtccccta 5940 gatccctcgg ctctgctctc ctttgcccat cagctccctg tggcctctat gctagtcctc 6000 cctattctcc tagtcaccct ttgcctctta agcccagcct tgactctcta catatttttc 6060 atttctcttt accctttagt gtacaatagt tgtagactct gacggaatga aaacaaacaa 6120 gaagcctaag cgttagaccc tgctgtggct aaagatctag aaggcatatc agttttctct 6180 gtcaggcttc ctacgttgcc ctcgtgcagg gctcattgct ctgcagagcg ctggaagccc 6240 cttcatgtat cgagagagaa gctggagcac agagctctga tgcacagttg gctgtttcag 6300 ctctgactca gggaggaaag aaggatgcca aggctctgtc acagcggaga agtctaatac 6360 cagagtggca cagagaagga ttccagggat cagtcagggt gtcctgtacc tatctgagta 6420 gagatgattc aaggatgggg acatattcca ggaaattgca caatcttccc ctggcagctt 6480 cagaggacga gtgaaaatag tgcagtatag tcaaagaagc aatgaattta ggatgaggaa 6540 cctgaagttt ctggaccagt ttggtcactg cttagttaca tgtccttagg aatgcttgtc 6600 acttaaccta aactttctgg gttttatcct cattgctcaa actaagcaac aacaccgaag 6660 cttctctaat ttgccttctg atgccaaatc ggtcaaaact gaggtcagca gctaccatga 6720 cgaaagtaaa atggtatatt gggtgaatgt ttgaaaattt cagattttat attctgtaaa 6780 gttatttact tactctattt caacaccaca aaatcttgaa atttggaagg taaccaagga 6840 agctttgaaa tttgttggca aacccggagt agtatggttt caaataccac tccaacccag 6900 cacctgaggt cgggcaaact cacaaaatag agtttctagg tagcaggtgt ttgctttggt 6960 tttgcaatag catttctcat aacagaactg aaactgtata gattacaaca ctccaaagtt 7020 attttttttt aacctaaaga accctacttc tagcacctcc tctgaaagaa tattttgctc 7080 caataccacg tgcaacaaac ttatttagac agtatcaact ctagccctta ataggttcaa 7140 ggacaagttt actgcttcct ttctacacgc ttaagttaca ctctcttctc ctgttctgat 7200 gactctaaat cttcagcctc tcccagcatt aatgtgtgag tcaacattgc agacctgaga 7260 aaggtttcta gaatggtaac ccaatgttct ttagaactgg agaattccat tttccttttc 7320 tggctttagt gttaatgtct gggaagggag gctcatgata agaaaataaa aatgagaaaa 7380 gaagatctag ttcataaaat gactataccc cacgtaagtt tgtaactgca cagtgttttg 7440 ttgtacggct cagaaatgag caagtgttag tattattagg cacacatcac gttcacagct 7500 tcgcgcagag acggatagtg tatttggata tatggaagtc atccaaatgc gttgctagaa 7560 tatcaggcct atgtttgttc tggtagaacc acagagactc acacaaagtt ctgaccctac 7620 cactctggtc tgggctgaga ccaccccaaa aaatcatcta gagaatcaaa ggggctcaca 7680 tctcagttgc attctattta aaagataaag gggccatact ctccactgca aagaagcatt 7740 agtaaaactt cccgcaccca aattataatc cggcaaccct agcttcagct aaaaagatta 7800 tccccagaat ctaaaagctt actattgttc tttcttggag tccaaaatgc ttagcagttc 7860 tgcagttcac atacaaaact caaaactctt tttatcctgt attctgtgag ttccttgatg 7920 atagtaattg ggtcttattc atcactctgt atcctgtggc tcacacagaa tctggcaaat 7980 tgcatatatt caataaatgt tgaatatcta ataacctcct tattaatata acaaatatta 8040 tcaacattct gtgcacatca atgtcccatg ctgctactgt agtcaggagt tatttggctg 8100 ggaagtggaa agtgttcaca aacatgatgc ttatctaata aaatatcact gagcaataag 8160 gagaaatatt ttaaatagat ttgaagttgt gaacaaataa tttagagtcc aaagaggaaa 8220 aagaaaatta actctgtttt ttatccctag aactcagaaa ctttactgga ttggtcaaca 8280 aagacaaact ttttattgta taaaacagta gaattcatgg aagggataat tcttttggaa 8340 caggcttctc ggttacttgc cttagtaaaa gaagtatctg gagggcctgg agagttcata 8400 aacatagttt tctgtgcaag tgaagatctg tagtttggct taaaaatcct tgcatgtagt 8460 tttgcttggc gtggaaagtt attaggagac aatcctttat cctgttctgc aagcaatcaa 8520 acacatgatc tcccttctag ccattcctca tccagtacat gggcatttcc agcctcctac 8580 atgtaacact tcaaacttcc tctgggcgtc tgcttagagc tatgtgatgg ccacagtttg 8640 ggcaaagccc ttctttcttt taaatatagg aactgttacc aagtttggtg caggcttgag 8700 gaataattac ataaaaatgg gcttccacga tggtgaatca gcatgctgat ttctcttggc 8760 aggaagctct cagggaaatg tctgtgaatc atttaaagat gctctctggt ggatagagaa 8820 accgcagtcc aaaaacagat gggtgtatat ttgctttgta agcaaagggc atgtatcata 8880 ctcgcatcta aaacagtgtc cacatctttg ccaaaggcac tgaattttac ctttttttat 8940 tacctcgtgg ccagcacttc ctcagtaggc atttcaggtg tgtcacaaac catgttcatt 9000 tttaatcttg aaaaagtcaa tcgtctgtag tctgtactat aaaatgatgc ctgcaacttt 9060 gtggtgaata ttatccccaa gttaacagtg aaataagtaa ataaaactat tattgaacaa 9120 a 9121 <110> SEOUL NATIONAL UNIVERSITY R&DB FOUNDATION <120> Method for promoting myelin plasticity <130> 21P10002 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 590 <212> PRT <213> Artificial Sequence <220> < 223> hM3Dq <400> 1 Met Thr Leu His Asn Asn Ser Thr Thr Ser Pro Leu Phe Pro Asn Ile 1 5 10 15 Ser Ser Ser Trp Ile His Ser Pro Ser Asp Ala Gly Leu Pro Pro Gly 20 25 30 Thr Val Thr His Phe Gly Ser Tyr Asn Val Ser Arg Ala Ala Gly Asn 35 40 45 Phe Ser Ser Pro Asp Gly Thr Thr Asp Asp Pro Leu Gly Gly His Thr 50 55 60 Val Trp Gln Val Val Phe Ile Ala Phe Leu Thr Gly Ile Leu Ala Leu 65 70 75 80 Val Thr Ile Ile Gly Asn Ile Leu Val Ile Val Ser Phe Lys Val Asn 85 90 95 Lys Gln Leu Lys Thr Val Asn Asn Tyr Phe Leu Leu Ser Leu Ala Cys 100 105 110 Ala Asp Leu Ile Ile Gly Val Ile Ser Met Asn Leu Phe Thr Thr Tyr 115 120 125 Ile Ile Met Asn Arg Trp Ala Leu Gly Asn Leu Ala Cys Asp Leu Trp 130 135 140 Leu Ala Ile Asp Cys Val Ala Ser Asn Ala Ser Val Met Asn Leu Leu 145 150 155 160 Val Ile Ser Phe Asp Arg Tyr Phe Ser Ile Thr Arg Pro Leu Thr Tyr 165 170 175 Arg Ala Lys Arg Thr Thr Lys Arg Ala Gly Val Met Ile Gly Leu Ala 180 185 190 Trp Val Ile Ser Phe Val Leu Trp Ala Pro Ala Ile Leu Phe Trp Gln 195 200 205 Tyr Phe Val Gly Lys Arg Thr Val Pro Pro Gly Glu Cys Phe Ile Gln 210 215 220 Phe Leu Ser Glu Pro Thr Ile Thr Phe Gly Thr Ala Ile Ala Gly Phe 225 230 235 240 Tyr Met Pro Val Thr Ile Met Thr Ile Leu Tyr Trp Arg Ile Tyr Lys 245 250 255 Glu Thr Glu Lys Arg Thr Lys Glu Leu Ala Gly Leu Gln Ala Ser Gly 260 265 270 Thr Glu Ala Glu Thr Glu Asn Phe Val His Pro Thr Gly Ser Ser Arg 275 280 285 Ser Cys Ser Ser Tyr Glu Leu Gln Gln Gln Ser Met Lys Arg Ser Asn 290 295 300 Arg Arg Lys Tyr Gly Arg Cys His Phe Trp Phe Thr Thr Lys Ser Trp 305 310 315 320 Lys Pro Ser Ser Glu Gln Met Asp Gln Asp His Ser Ser Ser Asp Ser 325 330 335 Trp Asn Asn Asn Asp Ala Ala Ala Ser Leu Glu Asn Ser Ala Ser Ser 340 345 350 Asp Glu Glu Asp Ile Gly Ser Glu Thr Arg Ala Ile Tyr Ser Ile Val 355 360 365 Leu Lys Leu Pro Gly His Ser Thr Ile Leu Asn Ser Thr Lys Leu Pro 370 375 380 Ser Ser Asp Asn Leu Gln Val Pro Glu Glu Glu Leu Gly Met Val Asp 385 390 395 400 Leu Glu Arg Lys Ala Asp Lys Leu Gln Ala Gln Lys Ser Val Asp Asp 405 410 415 Gly Gly Ser Phe Pro Lys Ser Phe Ser Lys Leu Pro Ile Gln Leu Glu 420 425 430 Ser Ala Val Asp Thr Ala Lys Thr Ser Asp Val Asn Ser Ser Val Gly 435 440 445 Lys Ser Thr Ala Thr Leu Pro Leu Ser Phe Lys Glu Ala Thr Leu Ala 450 455 460 Lys Arg Phe Ala Leu Lys Thr Arg Ser Gln Ile Thr Lys Arg Lys Arg 465 470 475 480 Met Ser Leu Val Lys Glu Lys Lys Ala Ala Gln Thr Leu Ser Ala Ile 485 490 495 Leu Leu Ala Phe Ile Ile Thr Trp Thr Pro Tyr Asn Ile Met Val Leu 500 505 510 Val Asn Thr Phe Cys Asp Ser Cys Ile Pro Lys Thr Phe Trp Asn Leu 515 520 525 Gly Tyr Trp Leu Cys Tyr Ile Asn Ser Thr Val Asn Pro Val Cys Tyr 530 535 540 Ala Leu Cys Asn Lys Thr Phe Arg Thr Thr Phe Lys Met Leu Leu Leu 545 550 555 560 Cys Gln Cys Asp Lys Lys Lys Arg Arg Lys Gln Gln Tyr Gln Gln Arg 565 570 575 Gln Ser Val Ile Phe His Lys Arg Ala Pro Glu Gln Ala Leu 580 585 590 <210> 2 <211> 9121 <212> DNA <213> Artificial Sequence <220> <223> hM3Dq <400> 2 acaccagctc tctcagtccg gggaggagga ggagcaggag gaacgcgagg agga aggaga 60 ggaggagcgg ccagcagtag ccacgaccgc caccaccagg cagaggaaga gttcgtgggg 120 aggaaaagac ctctcctccc ccttggagcg ccttctctct gcttttggag aaagggaata 180 cacgataaag aaggagacgg aaagaagaga aaaagtgagg cggggagacag agggaa aagg 240 gcgtgaacag aaagggccgg agcgtgcagg ggagcacagg gcgcgggggc ggcactgccg 300 agccgggagc gctgccgctt gggcaggtgc cgcggccgct gcccggaggc ggcatgtgac 360 gcgcggccgc agctgcccgc gggcggagcg ctctca gacc ccggagcgca caccgcgggg 420 ccatcggtgc catcgcggat ctccaggctc ctcatcagtc cgccggggcc gcagcagcgc 480 ttctgggaag acgggcgatg aactgaaggg cggctccggg caggggggca cgatcttaag 540 gacagtcgct ccctgaacgc ggagccggag gagacgaagg gaaggtggag cggacgccac 600 ccgcgcaccg ggcaggcgcg gagaccggcg tgggacagcc acctggagcg cagctgcc ag 660 atatgaacag taatggcaca tattttggtt atgagtcact cagaggactg tggattgaat 720 gaactgtatc catccccatc atgatgtaca gaaccaagtc tcttcactag ttttgctgtg 780 gcgtggcacc tggtctcttt ctagaaggaa agttcaacat acagcacaat tctggacaca 840 ttgtattggt ttgatgctcc tacctggaac agatcctcat gagctagtaa gtttgaaggg 900 gagtcataaa gaggaacagg cattatacgc aaggctgaag aagcagttgg taactaacag 960 tacatctttt tctaaggtgg agctggtctc ttgggcagcc tgacatctgg tccactcctc 1020 tgtcctatgc cgggatcatc atgaccgtag agattatgtc actgttttgc atccttgtta 1080 cataactcag ttcctggtag attactatgt cagagagtca caatgacctt gcacaataac 1140 agtacaacct cgcctttgtt tccaaacatc agctcctcct ggatacacag cccctccgat 1200 gcagggctgc ccccgggaac cgtcactcat ttcggcagct acaatgtttc tcgagcag ct 1260 ggcaatttct cctctccaga cggtaccacc gatgaccctc tgggaggtca taccgtctgg 1320 caagtggtct tcatcgcttt cttaacgggc atcctggcct tggtgaccat catcggcaac 1380 atcctggtaa ttgtgtcatt taaggtcaac aagcagctga agacggtcaa caactacttc 1440 ctcttaagcc tggcctgtgc cgatctgatt atcggggtca tttcaatgaa tctgtttacg 1500 acctacatca tcatgaatcg atgggcctta gggaacttgg cctgtgacct ctggcttgcc 1560 attgactacg tagccagcaa tgcctctgtt atgaatcttc tggtcatcag ctttgacaga 1620 tacttttcca tcacgaggcc gctcacgtac cgagccaaac gaacaacaaa gagagccggt 168 0 gtgatgatcg gtctggcttg ggtcatctcc tttgtccttt gggctcctgc catcttgttc 1740 tggcaatact ttgttggaaa gagaactgtg cctccgggag agtgcttcat tcagttcctc 1800 agtgagccca ccattacttt tggcacagcc atcgctgctt tttatatgcc tgtcaccatt 1860 atgactattt tatactggag gatctataag gaaactgaaa agcgtaccaa agagcttgct 1920 ggcctgcaag cctctggg ac agaggcagag acagaaaact ttgtccaccc cacgggcagt 1980 tctcgaagct gcagcagtta cgaacttcaa cagcaaagca tgaaacgctc caacaggagg 2040 aagtatggcc gctgccactt ctggttcaca accaagagct ggaaacccag ctccgagcag 2100 atggaccaag accac agcag cagtgacagt tggaacaaca atgatgctgc tgcctccctg 2160 gagaactccg cctcctccga cgaggaggac attggctccg agacgagagc catctactcc 2220 atcgtgctca agcttccggg tcacagcacc atcctcaact ccaccaagtt accctcatcg 2280 gacaacctgc aggtgcctga ggaggagctg gggatggtgg acttggagag gaaagccgac 2340 aagctgcagg cccagaagag cgtggacgat ggaggcagt t ttccaaaaag cttctccaag 2400 cttcccatcc agctagagtc agccgtggac acagctaaga cttctgacgt caactcctca 2460 gtgggtaaga gcacggccac tctacctctg tccttcaagg aagccactct ggccaagagg 2520 tttgctctga agaccagaag tcaga tcact aagcggaaaa ggatgtccct ggtcaaggag 2580 aagaaagcgg cccagaccct cagtgcgatc ttgcttgcct tcatcatcac ttggacccca 2640 tacaacatca tggttctggt gaacaccttt tgtgacagct gcatacccaa aaccttttgg 2700 aatctgggct actggctgg ctacatcaac agcaccgtga accccgtgg ctatgctctg 2760 tgcaacaaaa cattcagaac cactttcaag atgctgctgc tgtgccagtg tgacaaaaaa 2820 aagaggcgca agcagcagta ccagcagaga cagtcggtca tttttcacaa gcgcgcaccc 2880 gagcaggcct tgtagaatga ggttgtatca atagcagtga caaaacgcac acatcaaccc 2940 acagacctta ggaggaggaa gg cgagggcg gggtgacttc tggtgatgat aaaaatggtt 3000 ttatcaccca gatgtgaaag aagctgcctg tttactgatc cattgaataa acccatttta 3060 atagaaaaag tcaataccaa ttcagcaaaa agaaaaaaaa aacatactac tgaatataaa 3120 gaaatttatt ctgaaataga ctttacgtgt ttttttctta aagaggagaa aaatattgct 3180 tgacggcaat tatataccca aagtgatttg cctgggtcct a atagagctt cctgtctttt ctttggtgtg 3420 ttgttaaact ctatttgtgg acttgattct tgattcttgc aaagtactgt tttgtgcagt 3480 tcaagtttcg tacaaataaa aatacttaag tatatatata tgtgtgagtt ctgcacgcac 3540 acacatagtg tatataatat catgggaaac actgaactgg caaattattc ctgcaacata 3600 cgctttcagt actttggtaa ctgaagttct ctaggatcct aatgcaacat taacgtgaaa 36 60 taagcccagt gtaatgtttt tgaaaccagg gctgttttcc acagagagca gccaggcctt 3720 cccagcaggt ctgtgcagag cggacaggct cgtgagtcag ctgagcgccg tggcttcgcc 3780 agacttggtg ttaagcaacc tcctttgttg atgtctcaac agagctaa at cggggcccct 3840 ctgagctcaa agaatgaacc acatccacac gtttgaattt aatcatctaa atctgaatgt 3900 ttcagaacaa aatttctgct atctaaactg cttgaaactc aataatagtg tcacgtttga 3960 atgtcataca cagcaatata tatatatgg tatatatata tatatggcaa agcaaaaaaa 4020 aaaacatggt aagagagaat gaaggagaac attgtgtttg attcttgctg aatggcacct 4080 t ctcaaagaa aatagggctt gcacctttgt taatcagctg tggccagtgc tttctggtgt 4140 tcattgtgta accttcaccc aggaataggt gaggttttag gaagttacat gtcctctgaa 4200 gaaagaatta cactctgaaa agtaatgctt caaattgatt tccttacctt tt gggaaaaa 4260 aaaaaaattg tttttttgca ttctcccttg aattgaccaa aatgttaact gtttcatttg 4320 gggaggggat ggggtgctgc catcattgtc gttgttgttg ctgctgtagc tgttggggtt 4380 tctttcctg ttgccggggc tgtttgggga gagggagggg agggaggtgg gagggccgcg 4440 gagatatctt ccctttgtac agggcattcg tgttgtgaac ccagagctgg gtagaagctg 4500 cttttgtatt cagtgtgagg tggtgtttac agacgacttt gacaacagta gaagtgtact 4560 cagtggtgtc tgtgtatctg aactatttaa tttcgtgtta tgtttatatg cagaaatatt 4620 tatggatact acaccaagtg tttatttaat gttgataaat atgactcttc agt cgtcagc 4680 catggtgtcc tttcaaagtg attctttaag gtccacttga gcaatgaata gagtatattg 4740 gagctttcct gtggctaaga agaagaaaca tgtcatcctg ttggcatcac caagcaccta 4800 actctttcta ggtaataaaa agtcaacggt cacttcaacg taaatgttta aaaccactct 4860 ttagttaaat ctgatgggaa ctgaacaaag ctccaccatt gtgtctgaaa tcattttaca 4920 cagggaaaaa aaaaa acaat cagcataatt gtaagaatga tttttttggg cccacattca 4980 accaaagaac aagaaagaaa cagacgattt tctaatatgt acacagcaaa ctacagaatg 5040 tcatattatc cacctgtgaa acatctagta agcaattgca tctatatttt atttctcaag 5100 ttggcctggc acta ataaat atgtgaaagc ttattcaagt gtcccagtac ttcagcactt 5160 cccatccttt tggaaactcc tatcttgcct ccctcccaaa agggctgttt cttttattaa 5220 aaaaaaaaaa gaaatagaat taaatttgag aaaaggagaa accctgaaat ttaacagaac 5280 tttgagagtc acttccaatc tccttaaaca tgattatctc cttatccttg tccttttgtg 5340 atgctaataa tagcagaaaa ggaacaaatc caagactcta ggtctccctt ttttgtttta 5400 ctggttttat tatagaattg tttttaagcc tcttatggat gaaagctgca tttaaatact 5460 atctactgat tttatagaaa ggtatgtcca tttcttgggt cactttcaga gaaagaaaac 5520 aag caaaata ggttcttttg aatatgaata atttaaaagc ataattgaag atgtagtgag 5580 tatttataat tggaccttag ttacacgtgc atttctcaaa gggcttgtca ttacaggaga 5640 tattcagcac ttacctaaga tcaaaatgtc ttcatcccaa attgggaatc gaattatttc 5700 atgccatatg catgaggttt gcagacattt caagattagc cactttgctt gaagaagaa 5760 aatgatcaaa cctaaaataa actacacatg gagaatctct ctct ctctct ctgcctctct 5820 ctctctcttt ctccctcttc ttccttctct ctctctcaca agcacacaca cacacactca 5880 atctccattc tccaggcatt gccagatggt taggagaagt gagctctctc tggtccccta 5940 gatccctcgg ctctgctctc ctttgcccat cagct ccctg tggcctctat gctagtcctc 6000 cctattctcc tagtcaccct ttgcctctta agcccagcct tgactctcta catatttttc 6060 atttctcttt accctttagt gtacaatagt tgtagactct gacggaatga aaacaaacaa 6120 gaagcctaag cgttagaccc tgctgtggct aaagatctag aaggcatatc agttttctct 6180 gtcaggcttc ctacgttgcc ctcgtgcagg gctcattgct ctgcagagcg ctggaagccc 6240 cttcatgtat cgagagagaa gctggagcac agagctctga tgcacagttg gctgttttcag 6300 ctctgactca gggaggaaag aaggatgcca aggctctgtc acagcggaga agtctaatac 6360 cagagtggca cagagaagga ttccagggat cagtcagggt gtcct gtacc tatctgagta 6420 gagatgattc aaggatgggg acatattcca ggaaattgca caatcttccc ctggcagctt 6480 cagaggacga gtgaaaatag tgcagtatag tcaaagaagc aatgaattta ggatgaggaa 6540 cctgaagttt ctggaccagt ttggtcactg cttagttaca tgtccttagg aatgcttgtc 6600 acttaaccta aactttctgg gttttatcct cattgctcaa actaagcaac acaccgaag 6660 cttctctaat ttgccttctg atgccaaatc ggtcaaaact gaggtcagca gctaccatga 6720 cgaaagtaaa atggtatatt gggtgaatgt ttgaaaattt cagattttat attctgtaaa 6780 gttattact tactctattt caacaccaca aaatcttgaa atttggaa gg taaccaagga 6840 agctttgaaa tttgttggca aacccggagt agtatggttt caaataccac tccaacccag 6900 cacctgaggt cgggcaaact cacaaaatag agtttctagg tagcaggtgt ttgctttggt 6960 tttgcaatag catttctcat aacagaactg aaactgtata gattacaaca ctccaaagtt 7020 attttttttt aacctaaaga accctacttc tagcacctcc tctgaaagaa tattttgctc 708 agacc tgaga 7260 aaggtttcta gaatggtaac ccaatgttct ttagaactgg agaattccat tttccttttc 7320 tggctttagt gttaatgtct gggaagggag gctcatgata agaaaataaa aatgagaaaa 7380 gaagatctag ttcataaaat gactataccc cacgtaagtt tgtaactgca cagtgttttg 7440 ttgtacggct cagaaatgag caagtgttag tattattagg cacacatcac gttcacagct 7500 tcgcgca gag acggatagtg tatttggata tatggaagtc atccaaatgc gttgctagaa 7560 tatcaggcct atgtttgttc tggtagaacc acagagactc acacaaagtt ctgaccctac 7620 cactctggtc tgggctgaga ccaccccaaa aaatcatcta gagaatcaaa ggggctcaca 768 0 tctcagttgc attctattta aaagataaag gggccatact ctccactgca aagaagcatt 7740 agtaaaactt cccgcaccca aattataatc cggcaaccct agcttcagct aaaaagatta 7800 tccccagaat ctaaaagctt actattgttc tttcttggag tccaaaatgc ttagcagttc 7860 tgcagttcac atacaaaact caaaactctt tttatcctgt attctgtgag ttccttgatg 7920 atagtaatt g ggtcttattc atcactctgt atcctgtggc tcacacagaa tctggcaaat 7980 tgcatatatt caataaatgt tgaatatcta ataacctcct tattaatata acaaatatta 8040 tcaacattct gtgcacatca atgtcccatg ctgctactgt agtcaggagt tatttggctg 8100 ggaagtg gaa agtgttcaca aacatgatgc ttatctaata aaatatcact gagcaataag 8160 gagaaatatt ttaaatagat ttgaagttgt gaacaaataa tttagagtcc aaagaggaaa 8220 aagaaaatta actctgtttt ttatccctag aactcagaaa ctttactgga ttggtcaaca 8280 aagacaaact tttattgta taaaacagta gaattcatgg aagggataat tcttttggaa 8340 caggcttctc ggttacttgc cttagtaa aa gaagtatctg gagggcctgg agagttcata 8400 aacatagttt tctgtgcaag tgaagatctg tagtttggct taaaaatcct tgcatgtagt 8460 tttgcttggc gtggaaagtt attaggagac aatcctttat cctgttctgc aagcaatcaa 8520 acacatgatc tcccttctag ccattcctca tccagtacat gggcatttcc agcctcctac 8580 atgtaacact tcaaacttcc tctgggcgtc tgcttagagc tatgtgatgg ccacagtttg 8640 ggcaaagccc ttctttcttt taaatatag aactgttacc aagtttggtg caggcttgag 8700 gaataattac ataaaaatgg gcttccacga tggtgaatca gcatgctgat ttctcttggc 8760 aggaagctct cagggaaatg tctgtgaatc atttaaagat gctctctggt ggatagagaa 8820 accgcagtcc aaaaacagat gggtgtatat ttgctttgta agcaaagggc atgtatcata 8880 ctcgcatcta aaacagtgtc cacatctttg ccaaaggcac tgaattttac ctttttttat 8940 tacctcgtgg ccagcacttc ctcagtaggc atttcaggtg tgtcacaaac catgttcatt 9000 tttaatcttg aaaaagtcaa tcgtctgtag tctgtactat aaaatgatgc ctgcaacttt 9060 gtggtgaata ttatccccaa gttaacagtg aaataagtaa ataaaactat tattgaacaa 9120a 9121

Claims (8)

희소돌기신경교세포에 합성수용체를 발현시키는 단계; 상기 합성 수용체를 표적하는 합성 리간드를 처리하여 ERK1/2를 인산화시키는 단계를 포함하는 신경 수초의 가소성 증진 방법.
expressing a synthetic receptor in oligodendrocytes; A method for enhancing plasticity of a nerve sheath comprising the step of phosphorylating ERK1/2 by treating a synthetic ligand targeting the synthetic receptor.
청구항 1에 있어서, 인간을 제외한 동물에서 수행되는 것인 신경 수초의 가소성 증진 방법.
The method according to claim 1, which is performed in animals other than humans.
청구항 1에 있어서, 상기 합성수용체는 Gq-DREADD, Gs-DREADD 또는 β-arrestin preferring DREADD로 이루어진 군으로부터 선택되는 하나 이상인, 신경 수초의 가소성 증진 방법.
The method according to claim 1, wherein the synthetic receptor is at least one selected from the group consisting of Gq-DREADD, Gs-DREADD or β-arrestin preferring DREADD, the method for enhancing the plasticity of nerve sheath.
청구항 1에 있어서, 상기 합성 리간드는 클로자핀 N-옥사이드, 클로자핀 및 컴파운드-21로 이루어진 군으로부터 선택되는 하나 이상인, 신경 수초의 가소성 증진 방법.
The method according to claim 1, wherein the synthetic ligand is at least one selected from the group consisting of clozapine N-oxide, clozapine and compound-21, the plasticity enhancement method of nerve sheath.
희소돌기신경교세포를 표적하는 합성수용체를 포함하는 제1 제제; 및
상기 합성 수용체를 표적하는 합성 리간드를 포함하는 제2 제제를 포함하는 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물.
A first agent containing a synthetic receptor targeting oligodendrocytes; and
A composition for improving memory, exercise and cognitive abilities, comprising a second agent comprising a synthetic ligand targeting the synthetic receptor.
청구항 5에 있어서, 상기 합성수용체는 Gq-DREADD, Gs-DREADD 또는 β-arrestin preferring DREADD로 이루어진 군으로부터 선택되는 하나 이상인, 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물.
The composition according to claim 5, wherein the synthetic receptor is one or more selected from the group consisting of Gq-DREADD, Gs-DREADD or β-arrestin preferring DREADD.
청구항 5에 있어서, 상기 합성 리간드는 클로자핀 N-옥사이드, 클로자핀 및 컴파운드-21로 이루어진 군으로부터 선택되는 하나 이상인, 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물.
The composition according to claim 5, wherein the synthetic ligand is at least one selected from the group consisting of clozapine N-oxide, clozapine, and compound-21.
청구항 5에 있어서, 상기 제1 제제 및 제 2 제제는 동시에 또는 순차적으로 처리될 수 있는, 기억력 증진, 운동능력 향상 및 인지능력 개선용 조성물.The composition according to claim 5, wherein the first agent and the second agent can be processed simultaneously or sequentially.

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