JPH0676492B2 - Polyamino acid and method for producing the same - Google Patents
Polyamino acid and method for producing the sameInfo
- Publication number
- JPH0676492B2 JPH0676492B2 JP6790788A JP6790788A JPH0676492B2 JP H0676492 B2 JPH0676492 B2 JP H0676492B2 JP 6790788 A JP6790788 A JP 6790788A JP 6790788 A JP6790788 A JP 6790788A JP H0676492 B2 JPH0676492 B2 JP H0676492B2
- Authority
- JP
- Japan
- Prior art keywords
- polyamino acid
- side chain
- acid
- ester group
- general formula
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 239000002253 acid Substances 0.000 title claims description 40
- 238000004519 manufacturing process Methods 0.000 title claims description 8
- 125000004185 ester group Chemical group 0.000 claims description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 16
- 125000000217 alkyl group Chemical group 0.000 claims description 5
- 239000008346 aqueous phase Substances 0.000 claims description 4
- 125000003710 aryl alkyl group Chemical group 0.000 claims description 4
- 239000012670 alkaline solution Substances 0.000 claims description 3
- 229910052783 alkali metal Inorganic materials 0.000 claims description 2
- 150000001340 alkali metals Chemical class 0.000 claims description 2
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 2
- 239000000470 constituent Substances 0.000 claims 1
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 8
- 150000001413 amino acids Chemical class 0.000 description 8
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 7
- 238000010586 diagram Methods 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 238000007127 saponification reaction Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 235000013922 glutamic acid Nutrition 0.000 description 4
- 239000004220 glutamic acid Substances 0.000 description 4
- 239000002356 single layer Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 239000003513 alkali Substances 0.000 description 3
- 108700004370 poly-gamma-methylglutamate Proteins 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 230000000379 polymerizing effect Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 2
- 108010052285 Membrane Proteins Proteins 0.000 description 2
- 102000018697 Membrane Proteins Human genes 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- YEJSPQZHMWGIGP-UHFFFAOYSA-N dl-glutamic acid dimethyl ester Natural products COC(=O)CCC(N)C(=O)OC YEJSPQZHMWGIGP-UHFFFAOYSA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- SEWIYICDCVPBEW-UHFFFAOYSA-N methyl glutamate Chemical compound COC(=O)C(N)CCC(O)=O SEWIYICDCVPBEW-UHFFFAOYSA-N 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 238000005192 partition Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000013268 sustained release Methods 0.000 description 2
- 239000012730 sustained-release form Substances 0.000 description 2
- WVLBCYQITXONBZ-UHFFFAOYSA-N trimethyl phosphate Chemical compound COP(=O)(OC)OC WVLBCYQITXONBZ-UHFFFAOYSA-N 0.000 description 2
- JITSWUFGPFIMFG-UHFFFAOYSA-N 1,1,2,2,4-pentachlorobutane Chemical compound ClCCC(Cl)(Cl)C(Cl)Cl JITSWUFGPFIMFG-UHFFFAOYSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 239000000232 Lipid Bilayer Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 239000003012 bilayer membrane Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 125000004051 hexyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000012456 homogeneous solution Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 125000001165 hydrophobic group Chemical group 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 229920001308 poly(aminoacid) Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 229920005604 random copolymer Polymers 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
- Polyamides (AREA)
Description
【発明の詳細な説明】 〔産業上の利用分野〕 本発明は規則性の良い一次構造を有するポリアミノ酸お
よびその製造方法に関するものである。TECHNICAL FIELD The present invention relates to a polyamino acid having a highly ordered primary structure and a method for producing the same.
生体膜の持っている、特定の物質に対する選択透過性、
情報伝達、細胞間の相互認識といった機能は、膜タンパ
ク質と脂質二分子膜との相互作用に依存していると考え
られている。Selective permeability for specific substances possessed by biological membranes,
Functions such as information transmission and mutual recognition between cells are considered to depend on the interaction between the membrane protein and the lipid bilayer.
チャネルを形成するような膜タンパク質の一次構造は、
親・疎水性アミノ酸の周期的配列から成り、これがα−
ヘリックスを形成して二分子膜内で会合することによ
り、チャネル内部に親水性残基が、脂質側に疎水性残基
が配向した安定なチャネル構造が形成されると言われて
いる。The primary structure of a membrane protein that forms a channel is
It consists of a periodic sequence of parent and hydrophobic amino acids, which is α-
It is said that by forming a helix and associating in the bilayer membrane, a stable channel structure is formed in which hydrophilic residues are oriented inside the channel and hydrophobic residues are oriented on the lipid side.
一方、側鎖にエステル基を有するポリマーを、ケン化す
ることにより、カルボキシル基を有するポリマーに変換
することは一般に知られている。On the other hand, it is generally known that a polymer having an ester group in its side chain is converted into a polymer having a carboxyl group by saponification.
また、エステル基とカルボキシル基とを一分子中に有す
るポリマーは、これら側鎖を有するモノマーの重合によ
り得られることも一般に知られている。It is also generally known that a polymer having an ester group and a carboxyl group in one molecule can be obtained by polymerizing a monomer having these side chains.
膜タンパク質様物質として、規則性の良い一次構造を有
するポリアミノ酸を製造するために、前者の方法を用い
ると、側鎖エステル基を有するポリアミノ酸を、均一溶
液系でケン化することにより、エステル基を一定量のカ
ルボキシル基に変換することは可能であるが、一次構造
はランダムになってしまう。When the former method is used to produce a polyamino acid having a well-ordered primary structure as a membrane protein-like substance, the polyamino acid having a side chain ester group is saponified in a homogeneous solution system to give an ester. It is possible to convert the groups to a certain amount of carboxyl groups, but the primary structure becomes random.
又、後者の方法では、エステル基とカルボキシル基の量
比を一定の割合に調整することは出来るが、この二つの
基を規則正しく配列させること、即ち、分子の一次構造
を制御することは出来ず、ランダムな一次構造をもつも
のしか製造することはできなかった。Also, in the latter method, the amount ratio of the ester group and the carboxyl group can be adjusted to a certain ratio, but it is not possible to arrange these two groups regularly, that is, to control the primary structure of the molecule. However, only those having a random primary structure could be produced.
本発明は、エステル基側鎖を有するアミノ酸単位とカル
ボキシル基側鎖を有するアミノ酸単位とが規則正しく配
列した規則性の良い一次構造を有するポリアミノ酸およ
びその製造方法を提供することを目的とする。It is an object of the present invention to provide a polyamino acid having a regular primary structure in which an amino acid unit having an ester group side chain and an amino acid unit having a carboxyl group side chain are regularly arranged, and a method for producing the same.
すなわち、本発明は下記一般式（Ｉ）で示されるエステ
ル基側鎖の構造単位と下記一般式（II）で示されるカル
ボキシル基側鎖の構成単位とからなる、α−ヘリックス
構造を有するポリアミノ酸であって、該α−ヘリックス
構造による棒状体の軸方向特定の部分側面が、実質的に
一般式（II）で示されるカルボキシル基側鎖の構成単位
であることを特徴とするポリアミノ酸 （式中、ｍは１〜３の整数を表わし、Ｒはアルキル基ま
たはアラルキル基を表わす。） （式中、ｍは１〜３の整数を表わし、Ｍは水素原子又は
アルカリ金属を表わす。） および、α−ヘリックス構造を有し側鎖にエステル基を
含有するポリアミノ酸の単分子膜を水面上に形成した
後、水相にアルカリ溶液を添加して水中に存在するエス
テル基を選択的にケン化することを特徴とするポリアミ
ノ酸の製造方法を要旨とするものである。That is, the present invention is a polyamino acid having an α-helix structure, which comprises a structural unit of an ester group side chain represented by the following general formula (I) and a structural unit of a carboxyl group side chain represented by the following general formula (II). Wherein the partial side surface in the axial direction of the rod-shaped body having the α-helix structure is substantially a structural unit of a carboxyl side chain represented by the general formula (II). (In the formula, m represents an integer of 1 to 3, and R represents an alkyl group or an aralkyl group.) (In the formula, m represents an integer of 1 to 3, and M represents a hydrogen atom or an alkali metal.) And a monomolecular film of a polyamino acid having an α-helix structure and an ester group in its side chain is placed on the water surface. A gist of the present invention is to provide a method for producing a polyamino acid, characterized in that, after the above formation, an alkaline solution is added to an aqueous phase to selectively saponify ester groups present in water.
本発明に用いるα−ヘリックス構造を有し、側鎖にエス
テル基を含有するポリアミノ酸としては、前記一般式
（Ｉ）で示される繰返し単位からなるポリアミノ酸を挙
げることができる。Examples of the polyamino acid having an α-helix structure used in the present invention and having an ester group in its side chain include a polyamino acid comprising a repeating unit represented by the above general formula (I).
一般式（Ｉ）において、ｍは１〜３の整数、Ｒはアルキ
ル基またはアラルキル基を表わすが、アルキル基として
は、メチル、エチル、プロピル、ヘキシル、オクチル等
及び炭素数18程度までの長鎖アルキル基が好ましく挙げ
られ、またアラルキル基としてはベンジル基等が好まし
く挙げられる。これらのうち、特に好ましいのはメチ
ル、エチル基である。In the general formula (I), m represents an integer of 1 to 3 and R represents an alkyl group or an aralkyl group, and the alkyl group is methyl, ethyl, propyl, hexyl, octyl or the like and a long chain having up to about 18 carbon atoms. An alkyl group is preferable, and a benzyl group is preferable as the aralkyl group. Of these, methyl and ethyl groups are particularly preferable.
一般式（Ｉ）を与えるモノアミノ酸としては、グルタミ
ン酸、アスパラギン酸が挙げられ、一般式（Ｉ）の単位
からなるポリアミノ酸としては、ポリ（Ｌ−グルタミン
酸−γ−ベンジル）、ポリ（Ｌ−アスパラギン酸−β−
ベンジル）、好ましくはポリ（Ｌ−グルタミン酸−γ−
メチル）、ポリ（Ｌ−グルタミン酸−γ−エチル）、ポ
リ（Ｌ−アスパラギン酸−β−メチル）、ポリ（Ｌ−ア
スパラギン酸−β−エチル）等が挙げられる。Glutamic acid and aspartic acid may be mentioned as the monoamino acid giving the general formula (I), and poly (L-glutamic acid-γ-benzyl) and poly (L-asparagine) may be mentioned as the polyamino acid comprising the unit of the general formula (I). Acid-β-
Benzyl), preferably poly (L-glutamic acid-γ-
Methyl), poly (L-glutamic acid-γ-ethyl), poly (L-aspartic acid-β-methyl), poly (L-aspartic acid-β-ethyl) and the like.
これらポリアミノ酸は、α−アミノ酸−Ｎ−無水物（い
わゆるNCA）の重合により得られる。例えば、ポリ（Ｌ
−グルタミン酸−γ−メチル）は、γ−メチル−Ｌ−グ
ルタメート−Ｎ−カルボン酸無水物を、有機アミン、ア
ルコラート、水、アルカリ等の触媒を用いて公知の方法
で重合することにより得られる。These polyamino acids are obtained by polymerizing α-amino acid-N-anhydride (so-called NCA). For example, poly (L
-Glutamic acid-γ-methyl) is obtained by polymerizing γ-methyl-L-glutamate-N-carboxylic acid anhydride by a known method using a catalyst such as an organic amine, an alcoholate, water and an alkali.
単分子膜の製造は、常法（例えば、新実験化学講座18巻
「界面とコロイド」（丸善、昭和52年10月）p.442）に
よることができる。例えば、前記ポリアミノ酸を、非水
系でかつポリアミノ酸と非会合である溶媒（好ましく
は、クロロホルム、トリクロロエチレン、エチレンジク
ロライド、ベンゼン等）に、溶解して0.01〜１％溶液を
調製し、これを水面上に展開し、膜が固体状態となる表
面圧で調整し、α−ヘリックス構造を有するポリアミノ
酸の単分子膜が、形成される。The monomolecular film can be produced by a conventional method (for example, New Experimental Chemistry Course, Vol. 18, "Interface and Colloid" (Maruzen, October 1977), p. 442). For example, the polyamino acid is dissolved in a solvent that is non-aqueous and does not associate with the polyamino acid (preferably chloroform, trichloroethylene, ethylene dichloride, benzene, etc.) to prepare a 0.01 to 1% solution, which is dissolved on the water surface. A monomolecular film of polyamino acid having an α-helix structure is formed by adjusting the surface pressure so that the film expands to a solid state.
次に、図面に従って、本発明を詳細に説明すると、第１
図は本発明の製造方法の各工程を示す図である。水槽１
に水を入れ、仕切り板２を移動させて清浄な水面３とす
る。次にポリアミノ酸の上記溶液をマイクロシリンジ４
を用いて一定量を水面３にゆっくり滴下し、ポリアミノ
酸単分子５の膜を展開し仕切り板２を移動させて単分子
膜６を作成する（１−ａ図）。Next, the present invention will be described in detail with reference to the drawings.
The figure is a diagram showing each step of the manufacturing method of the present invention. Aquarium 1
Water is put in and the partition plate 2 is moved to obtain a clean water surface 3. Next, add the above solution of polyamino acid to the microsyringe 4
Then, a fixed amount is slowly dropped onto the water surface 3 by using, the membrane of the polyamino acid monomolecule 5 is developed, and the partition plate 2 is moved to form the monomolecular membrane 6 (Fig. 1-a).
この単分子膜を形成するポリアミノ酸５は、α−ヘリッ
クス構造を有するので、側鎖エステル基はヘリックス上
に100゜の間隔をおいて存在する。即ち、エステル基の
配列は、そのヘリックス断面に対する投影図（第２図）
の如く、エステル基18個で５回転（3.6個／回転）が単
位周期である。ただし第２図中の番号は分子鎖末端アミ
ノ酸の側鎖エステル基を１として示した。単分子膜形成
時においてエステル基が水側に存在するか非水（空気）
側に在るかはヘリックス断面に対する水のぬれ角度に依
存するが、これを120゜に採ると、ヘリックス上のエス
テル基は概略水側のエステル基−空気側のエステル基
−空気側のエステル基のくり返しで形成される。Since the polyamino acid 5 forming this monomolecular film has an α-helix structure, side chain ester groups are present on the helix at intervals of 100 °. That is, the array of ester groups is a projected view of the cross section of the helix (Fig. 2).
As described above, the unit cycle is 5 rotations (3.6 rotations / rotation) with 18 ester groups. However, the numbers in FIG. 2 indicate the side chain ester group of the amino acid at the terminal of the molecular chain as 1. Does the ester group exist on the water side during monolayer formation? Non-water (air)
Whether it is on the side depends on the wetting angle of water with respect to the cross section of the helix, but if this is taken at 120 °, the ester group on the helix is roughly the ester group on the water side-ester group on the air side-ester group on the air side. It is formed by repeating.
次いでエステル基の1/2モル以上の量のアルカリを含む
水溶液を、１−ｂ図の如くマイクロシリンジ４等で添加
し、単分子膜６の水相側をケン化させる。ケン化時間
は、アルカリ溶液が均一分散後５〜10分間で十分であ
る。ケン化反応後、アルカリと等モル量の酸を添加し、
反応を停止させる（１−ｃ図）。酸としては、塩酸、リ
ン酸、酢酸等を挙げることができる。Then, an aqueous solution containing an alkali in an amount of 1/2 mole or more of the ester group is added by a microsyringe 4 or the like as shown in FIG. 1-b to saponify the aqueous phase side of the monomolecular film 6. The saponification time is 5 to 10 minutes after the alkaline solution is uniformly dispersed. After the saponification reaction, add an equimolar amount of acid to the alkali,
Stop the reaction (Fig. 1-c). Examples of the acid include hydrochloric acid, phosphoric acid, acetic acid and the like.
単分子膜製造から中和に至る工程においての水温として
は、５〜30℃を挙げることができる。The water temperature in the steps from monolayer production to neutralization may be 5 to 30 ° C.
かくして、水中に存在するエステル基のみ、即ち、単分
子膜の片側のみがケン化され、概略カルボキシル基１ケ
にエステル基２ケのくり返しからなる、規則性のよい一
次構造を有するポリアミノ酸が得られる。Thus, a polyamino acid having a well-ordered primary structure is obtained, in which only the ester group present in water, that is, only one side of the monolayer is saponified, and approximately one carboxyl group and two ester groups are repeated. To be
１−ｄ図は、本発明のポリアミノ酸の構造を示す模式図
であり、５はα−ヘリックス構造による棒状体として表
わされるポリアミノ酸分子を示し、６はケン化されてい
ないエステル基側鎖部であり、斜線部７はケン化された
カルボキシル基側鎖部であり、カルボキシル基側鎖部が
α−ヘリックス構造による棒状体の軸方向部分側面に存
在することを示す。1-d is a schematic view showing the structure of the polyamino acid of the present invention, 5 shows a polyamino acid molecule represented as a rod-like body having an α-helix structure, and 6 shows an unsaponified ester group side chain portion. And the shaded portion 7 is a saponified carboxyl group side chain portion, and indicates that the carboxyl group side chain portion exists on the side surface in the axial direction of the rod-shaped body having the α-helix structure.
エステル基のケン化可能な範囲がヘリックス断面に対し
120゜の範囲であると仮定すると、次の18個のアミノ酸
配列をくり返し単位とする規則性のよい一次構造を有す
るポリアミノ酸が得られる。The saponifiable range of ester groups is
Assuming a range of 120 °, a polyamino acid having a regular primary structure having the following 18 amino acid sequences as repeating units is obtained.
EAEEAAEEAEEEAEEAAEｎ （ここでＥおよびＡは、各々前記一般式（Ｉ）のエステ
ル基、一般式（II）のカルボキシル基を有するアミノ酸
単位を示す。）この場合、重合度30のポリアミノ酸に対
するカルボキシル基の含率は36.7％となる。同様にケン
化可能な範囲を100゜、140゜、160゜に仮定した時のカ
ルボキシル基含率は、各々30％、40％、46.7％となる。EAEEAAEEAEEEAEEAAEn (wherein E and A represent an amino acid unit having an ester group of the general formula (I) and a carboxyl group of the general formula (II), respectively). In this case, a polyamino acid having a degree of polymerization of 30 contains a carboxyl group. The rate is 36.7%. Similarly, assuming that the saponifiable range is 100 °, 140 °, and 160 °, the carboxyl group contents are 30%, 40%, and 46.7%, respectively.
本発明で得られるポリアミノ酸は、両親媒性であり、二
分子膜形成物質に膜タンパク質様物質として添加する
と、有効なチャネルを形成することが出来る。従って、
徐放薬剤等への応用が考えられる。The polyamino acid obtained in the present invention is amphipathic and can form an effective channel when added to a bilayer membrane-forming substance as a membrane protein-like substance. Therefore,
Application to a sustained-release drug, etc. is considered.
以下、実施例により本発明を更に詳細に説明するが、本
発明は以下の実施例に限定されるものではない。Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to the following Examples.
実施例１ ＜ポリアミノ酸溶液の調製＞ γ−メチル−Ｌ−グルタメート−Ｎ−カルボン酸をクロ
ロホルムに溶解して10％溶液とし、これにトリエチルア
ミンを加え室温で重合したところ、重合度30の低分子量
ポリ（γ−メチル−Ｌ−グルタメート）を得た。これ
を、クロロホルムで希釈し、0.1％濃度に調製した。Example 1 <Preparation of polyamino acid solution> [gamma] -methyl-L-glutamate-N-carboxylic acid was dissolved in chloroform to make a 10% solution, and triethylamine was added thereto and polymerized at room temperature. Poly (γ-methyl-L-glutamate) was obtained. This was diluted with chloroform to prepare a 0.1% concentration.
＜単分子膜の製造＞ 前記ポリ（γ−メチル−Ｌ−グルタメート）の0.1％ク
ロロホルム溶液を、室温下水面上に展開し、膜が固体状
態となる表面圧（第３図）で調整し、α−ヘリックス構
造を有するポリアミノ酸の単分子膜を製造した。<Production of Monomolecular Film> A 0.1% chloroform solution of the poly (γ-methyl-L-glutamate) was developed on the water surface at room temperature and adjusted with the surface pressure (FIG. 3) at which the film becomes a solid state, A polyamino acid monolayer having an α-helix structure was produced.
＜単分子膜の片側面のケン化方法＞ 室温下で、水相の水酸化カリウム濃度が0.1モル（メチ
ルグルタメート総残基の105倍モル）となるように、水
酸化カリウム溶液を添加し、３分間ケン化反応を行なっ
た。その後等量の塩酸で反応を停止し、目的の片側ケン
化の膜を得た。<Saponification method on one side of monomolecular film> At room temperature, a potassium hydroxide solution was added so that the concentration of potassium hydroxide in the aqueous phase was 0.1 mol (10 5 times mol of total residues of methyl glutamate). Saponification reaction was performed for 3 minutes. After that, the reaction was stopped with an equal amount of hydrochloric acid to obtain a desired one-side saponified membrane.
＜ケン化膜の評価＞ 得られた片側ケン化膜のトリフルオロ酢酸系での高分解
能NMRスペクトルのα−水素と側鎖メトキシのピークの
面積の比よりグルタミン酸コンテントは大過剰のKOH添
加にもかかわらず34モル％であった。またこの値はヘリ
ックス断面に対するケン化可能な範囲を120゜として得
たグルタミン酸コンテントの計算値36.7％と良好に一致
した。<Evaluation of saponified film> From the ratio of the peak areas of α-hydrogen and side chain methoxy in the high-resolution NMR spectrum of the obtained one-sided saponified film in trifluoroacetic acid system, glutamic acid content can be added even in a large excess of KOH. It was 34 mol% regardless. This value was in good agreement with the calculated value of 36.7% for glutamic acid content obtained by setting the saponifiable range to the helix cross section at 120 °.
第４図に、本願発明の片側ケン化物（Ａ）、比較例とし
てメチルグルタメート／グルタミン酸が70/30のランダ
ム共重合体（Ｂ）およびポリ（γ−メチル−Ｌ−グルタ
メート）（Ｃ）の側鎖NMRを測定した結果を示す。三種
のスペクトルの差異は明白であり、片側ケン化物（Ａ）
の低磁場側の複雑なショルダーは、このポリアミノ酸の
一次構造の規則性を示している。FIG. 4 shows the side of the one-sided saponified product (A) of the present invention, a random copolymer (B) of 70/30 methylglutamate / glutamic acid and poly (γ-methyl-L-glutamate) (C) as comparative examples. The result of having measured chain NMR is shown. The difference in the spectra of the three species is clear, and the one-sided saponified product (A)
The intricate shoulder on the low magnetic field side of γ indicates the regularity of the primary structure of this polyamino acid.
また、本発明の片側ケン化ポリアミノ酸の二次構造をCD
測定（JASCO J−40C）によって検討した（第５図）。リ
ン酸トリメチル（TMP）系、水系（pH7.2）およびベシク
ル系（pH7.2）のいずれにも可溶であり、222nmおよび20
8nmに負のピークを持つα−ヘリックス構造に固有のス
ペクトルを示し、水系およびベシクル系でのヘリックス
コンテントは約40％となった。以上のことから考える
と、ポリメチルグルタメートを単分子膜状態でケン化し
て得たポリアミノ酸は、両親媒性物質であり、pH7.2に
おいてもヘリックス構造を形成しうることが判った。In addition, the secondary structure of the unilaterally saponified polyamino acid of the present invention is determined by the CD
It was examined by measurement (JASCO J-40C) (Fig. 5). Soluble in trimethyl phosphate (TMP) system, water system (pH7.2) and vesicle system (pH7.2), 222nm and 20nm
The spectrum unique to the α-helix structure with a negative peak at 8 nm was shown, and the helix content in water and vesicle systems was about 40%. From the above, it was found that the polyamino acid obtained by saponifying polymethylglutamate in a monomolecular film state is an amphipathic substance and can form a helix structure even at pH 7.2.
本発明方法によれば、エステル基側鎖を有するアミノ酸
単位とカルボキシル基側鎖を有するアミノ酸単位とが規
則正しく配列した規則性の良い一次構造を有するポリア
ミノ酸を得ることができる。According to the method of the present invention, it is possible to obtain a polyamino acid having a regular primary structure in which an amino acid unit having an ester group side chain and an amino acid unit having a carboxyl group side chain are regularly arranged.
又、本発明のポリアミノ酸は、物質の選択透過等に有用
であり、例えば、徐放薬剤等に応用可能である。Further, the polyamino acid of the present invention is useful for selective permeation of substances and the like, and can be applied to, for example, sustained-release drugs.
第１図は本発明のポリアミノ酸の製造工程を示す図、第
２図は側鎖エステル基の配列を示す模式図、第３図はポ
リ（Ｌ−グルタメート−γ−メチル）の単分子膜が固体
状態となる表面圧を示す図、第４図は本発明のポリアミ
ノ酸のNMRスペクトルを示す図、第５図は本発明のポリ
アミノ酸の二次構造をCD測定した結果を示す図である。FIG. 1 is a diagram showing a process for producing a polyamino acid of the present invention, FIG. 2 is a schematic diagram showing a sequence of side chain ester groups, and FIG. 3 shows a poly (L-glutamate-γ-methyl) monomolecular film. FIG. 4 is a diagram showing the surface pressure in the solid state, FIG. 4 is a diagram showing the NMR spectrum of the polyamino acid of the present invention, and FIG. 5 is a diagram showing the results of CD measurement of the secondary structure of the polyamino acid of the present invention.
Claims (2)
鎖の構成単位と下記一般式（II）で示されるカルボキシ
ル基側鎖の構成単位とからなる、α−ヘリックス構造を
有するポリアミノ酸であって、該α−ヘリックス構造に
よる棒状体の軸方向特定の部分側面が、実質的に一般式
（II）で示されるカルボキシル基側鎖の構成単位である
ことを特徴とするポリアミノ酸。 （式中、ｍは１〜３の整数を表わし、Ｒはアルキル基ま
たはアラルキル基を表わす。） （式中、ｍは１〜３の整数を表わし、 Ｍは水素原子又はアルカリ金属を表わす。）1. A polyamino acid having an α-helix structure, which comprises a structural unit of an ester group side chain represented by the following general formula (I) and a structural unit of a carboxyl group side chain represented by the following general formula (II). A polyamino acid characterized in that the side surface in the axial direction of the rod-shaped body having the α-helix structure is a constituent unit of the carboxyl group side chain substantially represented by the general formula (II). (In the formula, m represents an integer of 1 to 3, and R represents an alkyl group or an aralkyl group.) (In the formula, m represents an integer of 1 to 3, and M represents a hydrogen atom or an alkali metal.)
基を含有するポリアミノ酸の単分子膜を水面上に形成し
た後、水相にアルカリ溶液を添加して水中に存在するエ
ステル基を選択的にケン化することを特徴とするポリア
ミノ酸の製造方法。2. After forming a monomolecular film of a polyamino acid having an α-helix structure and containing an ester group on its side chain on the water surface, an alkaline solution is added to the aqueous phase to remove the ester group present in water. A method for producing a polyamino acid, which comprises selectively saponifying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP6790788A JPH0676492B2 (en) | 1988-03-22 | 1988-03-22 | Polyamino acid and method for producing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP6790788A JPH0676492B2 (en) | 1988-03-22 | 1988-03-22 | Polyamino acid and method for producing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH01240524A JPH01240524A (en) | 1989-09-26 |
| JPH0676492B2 true JPH0676492B2 (en) | 1994-09-28 |
Family
ID=13358441
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP6790788A Expired - Lifetime JPH0676492B2 (en) | 1988-03-22 | 1988-03-22 | Polyamino acid and method for producing the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0676492B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02214726A (en) * | 1989-02-15 | 1990-08-27 | Fuji Photo Film Co Ltd | Polypeptide thin film and production of material carrying same |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5031906B2 (en) | 2008-01-30 | 2012-09-26 | カーディアック ペースメイカーズ， インコーポレイテッド | Radiation effect detection method and apparatus |
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1988
- 1988-03-22 JP JP6790788A patent/JPH0676492B2/en not_active Expired - Lifetime
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5031906B2 (en) | 2008-01-30 | 2012-09-26 | カーディアック ペースメイカーズ， インコーポレイテッド | Radiation effect detection method and apparatus |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH01240524A (en) | 1989-09-26 |
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