JP5142313B2 - Polysarcosine derivative and drug carrier comprising the same as a membrane constituent - Google Patents

Description

  The present invention relates to a novel polysarcosine derivative and a drug carrier containing this as one of membrane constituents.

  In recent years, research on drug delivery systems (DDS) that efficiently distribute drugs to target organs has become active. For example, a method of using closed vesicles such as liposomes, emulsions, lipid microspheres, and nanoparticles as a drug carrier, a method of including or binding a drug to a polymer carrier such as a polymer synthetic polymer micelle or polysaccharide, These closed vesicles and polymer carriers can be used to improve the target directivity by modifying the surface with high-molecular functional molecules such as antibodies and proteins, and low-molecular functional molecules such as specific sugar chains and peptides. (See Non-Patent Document 1 and Non-Patent Document 2).

  However, when these drug carriers (hereinafter also referred to as “drug carriers”) are put to practical use, there are various problems to be overcome. Among them, avoidance from the foreign body recognition mechanism on the living body side and control of pharmacokinetics are difficult. Is a problem. In particular, closed vesicles are aggregated by interaction with opsonin proteins and plasma proteins in the blood and captured by reticuloendothelial tissues (RES) such as the liver and spleen. It was a difficult situation for highly selective delivery.

  As means for solving the above problems, plasma proteins, opsonin proteins, and the like can be obtained by coating the surface of a polymer carrier such as these closed vesicles with a hydrophilic polymer such as polyethylene glycol (PEG). Adsorption is prevented to increase blood stability (retention in blood), and capture by RES can be avoided.

However, it is known that low molecular weight PEG is highly safe and is easily excreted from the kidney. However, high molecular weight PEG derivatives (eg, polyethylene glycol) surface-coated on such closed vesicles and the like. -Metabolism / degradation of PEG after phosphatidylethanolamine (PEG-PE)) has been taken up into cells has not been clarified, and safety is not necessarily ensured.
Therefore, even if it has a high molecular weight, a novel compound that itself is biodegradable, safer, has high blood stability (retention in blood), and can be prevented from being captured by RES is highly desired. ing.

Sarcosine is an amino acid that exists in various animal and plant tissues and is converted to glycine by oxidative demethylation by sarcosine dehydrogenase. A polymer of sarcosine is known as polysarcosine.
However, long-chain alkyl derivatives of polysarcosine are not known.

Cancer Letters, USA, 1997, Vol. 118, No. 2, p. 153 British Journal of Cancer, UK, 1997, Vol. 76, No. 1, p. 83

  Therefore, the present invention provides a drug carrier effective as a DDS preparation that can efficiently and safely target a drug to a target site, and a novel polysarcosine derivative that can be used as a membrane component of the drug carrier. The purpose is to provide.

（但し、式（１）中、Ｒ^１は水素原子、アミノ基の保護基またはアミノ基に結合する官能基であり、Ｒ^２およびＲ^３は互いに独立に炭素数１０〜２５のアルキル基またはアルケニル基である。lは５〜１５０の整数を示す）
（２）下記一般式（２）で示されるポリサルコシン誘導体。

（但し、式（２）中、Ｒ^４は水素原子、アミノ基の保護基またはアミノ基に結合する官能基であり、Ｒ^５およびＲ^６は互いに独立に炭素数１０〜２５のアルキル基またはアルケニル基である。ｋは１０〜１００の整数を示す。ｍおよびｎは互いに独立に０または１〜３の整数を示す）
（３）前記一般式（１）おいて、Ｒ^２およびＲ^３は互いに独立に炭素数１２〜１８のアルキル基またはアルケニル基である（１）に記載のポリサルコシン誘導体。
（４）前記一般式（２）おいて、Ｒ^５およびＲ^６は互いに独立に炭素数１２〜１８のアルキル基またはアルケニル基である（２）に記載のポリサルコシン誘導体。
（５）前記一般式（１）において、Ｒ^２およびＲ^３は炭素数１８のアルキル基であり、ｌは２０〜６０である（１）または（３）に記載のポリサルコシン誘導体。
（６）前記一般式（２）において、Ｒ^５およびＲ^６は炭素数１８のアルキル基であり、ｋは２０〜６０である（２）または（４）に記載のポリサルコシン誘導体。
（７）（１）ないし（６）のいずれかに記載のポリサルコシン誘導体を膜構成成分の一つとして含有する薬物担体。
（８）前記薬物担体が、閉鎖小胞である（７）に記載の薬物担体。
（９）さらに、リン脂質、リン脂質以外のグリセロ脂質、スフィンゴ脂質およびコレステロール類からなる群から選ばれる少なくとも一つを含有する（７）または（８）に記載の薬物担体。
（１０）さらに、安定化剤および酸化防止剤からなる群から選ばれる少なくとも一つを含有する（７）ないし（９）に記載のいずれかの薬物担体。
（１１）担持される薬物が、核酸、ポリヌクレオチド、遺伝子およびそれを含むベクター、抗癌剤、抗生物質、酵素剤、酵素阻害剤、抗酸化剤、脂質取り込み阻害剤、ホルモン剤、抗炎症剤、ステロイド剤、血管拡張剤、アンジオテンシン変換酵素阻害剤、アンジオテンシン受容体拮抗剤、平滑筋細胞の増殖および／または遊走阻害剤、血小板凝集阻害剤、抗凝固剤、ケミカルメディエーターの遊離抑制剤、血管内皮細胞の増殖または抑制剤、アルドース還元酵素阻害剤、メサンギウム細胞増殖阻害剤、リポキシゲナーゼ阻害剤、免疫抑制剤、免疫賦活剤、抗ウイルス剤、アミロイドーシス阻害剤、ＮＯＳ阻害剤、ＡＧＥｓ（Ａｄｖａｎｃｅｄ ｇｌｙｃａｔｉｏｎ ｅｎｄｐｒｏｄｕｃｔｓ）阻害剤、ラジカルスカベンジャー、グリコサミノグリカンおよびその誘導体、オリゴ糖および／または多糖およびそれらの誘導体、タンパク質およびペプチドからなる群から選択される少なくとも一つの予防または治療薬である（７）ないし（１０）のいずれかに記載の薬物担体。 In order to solve the above problems, the present invention provides the following (1) to (19).
(1) A polysarcosine derivative represented by the following general formula (1).

(In the formula (1), R ¹ is a hydrogen atom, an amino-protecting group or a functional group bonded to an amino group, and R ² and R ³ are each independently an alkyl group or alkenyl having 10 to 25 carbon atoms. (L represents an integer of 5 to 150)
(2) A polysarcosine derivative represented by the following general formula (2).

(In the formula (2), R ⁴ is a hydrogen atom, an amino protecting group, or a functional group bonded to an amino group, and R ⁵ and R ⁶ are each independently an alkyl or alkenyl having 10 to 25 carbon atoms. (K represents an integer of 10 to 100. m and n each independently represents an integer of 0 or 1 to 3)
(3) The polysarcosine derivative according to (1), wherein in the general formula (1), R ² and R ³ are each independently an alkyl group or an alkenyl group having 12 to 18 carbon atoms.
(4) The polysarcosine derivative according to (2), wherein, in the general formula (2), R ⁵ and R ⁶ are each independently an alkyl group or an alkenyl group having 12 to 18 carbon atoms.
(5) The polysarcosine derivative according to (1) or (3), wherein, in the general formula (1), R ² and R ³ are an alkyl group having 18 carbon atoms, and 1 is 20 to 60.
(6) The polysarcosine derivative according to (2) or (4), wherein, in the general formula (2), R ⁵ and R ⁶ are alkyl groups having 18 carbon atoms, and k is 20 to 60.
(7) A drug carrier comprising the polysarcosine derivative according to any one of (1) to (6) as one of membrane constituents.
(8) The drug carrier according to (7), wherein the drug carrier is a closed vesicle.
(9) The drug carrier according to (7) or (8), further comprising at least one selected from the group consisting of phospholipids, glycerolipids other than phospholipids, sphingolipids, and cholesterols.
(10) The drug carrier according to any one of (7) to (9), further comprising at least one selected from the group consisting of a stabilizer and an antioxidant.
(11) The drug to be carried is a nucleic acid, a polynucleotide, a gene and a vector containing the same, an anticancer agent, an antibiotic, an enzyme agent, an enzyme inhibitor, an antioxidant, a lipid uptake inhibitor, a hormone agent, an anti-inflammatory agent, a steroid Agent, vasodilator, angiotensin converting enzyme inhibitor, angiotensin receptor antagonist, smooth muscle cell proliferation and / or migration inhibitor, platelet aggregation inhibitor, anticoagulant, chemical mediator release inhibitor, vascular endothelial cell inhibitor Proliferation or inhibitor, aldose reductase inhibitor, mesangial cell proliferation inhibitor, lipoxygenase inhibitor, immunosuppressant, immunostimulator, antiviral agent, amyloidosis inhibitor, NOS inhibitor, AGEs (Advanced glycation endproducts) inhibitor, Radical scavenger, glycosamino The drug carrier according to any one of (7) to (10), which is at least one prophylactic or therapeutic agent selected from the group consisting of ricane and derivatives thereof, oligosaccharides and / or polysaccharides and derivatives thereof, proteins and peptides .

(12) The carried drug is at least one in-vivo diagnostic agent selected from the group consisting of an X-ray contrast agent, an ultrasound diagnostic agent, a radioisotope-labeled nuclear medicine diagnostic agent, and a diagnostic agent for nuclear magnetic resonance diagnosis ( 7) The drug carrier according to any one of (10).
(13) A pharmaceutical composition comprising the drug carrier according to any one of (7) to (12).

(14) A method for delivering a drug carried on a drug carrier to a target site, comprising administering the drug carrier according to any one of (7) to (12) to a host.
(15) A method for transporting a drug carrier to a target site, comprising administering the drug carrier according to any one of (7) to (12) to a host.
(16) A method for preventing or treating a disease, comprising administering an effective amount of the drug carrier according to any one of (7) to (11) to a host.

(17) A method for diagnosing a disease, comprising administering an effective amount of the drug carrier according to any one of (7) to (10) or (12) to a host.
(18) A composition for delivering a drug carried on a drug carrier to a target site, comprising an effective amount of the drug carrier according to any one of (7) to (12).
(19) A composition for transporting a drug carrier to a target site, comprising an effective amount of the drug carrier according to any one of (7) to (12).
In addition, the “effective amount” described in (16) to (19) means the amount of the drug carrier that has an amount that is effective for prevention, treatment, or diagnosis described later. To do.

According to the present invention, a novel polysarcosine derivative that can be used as a constituent of a drug carrier is provided.
The polysarcosine derivative of the present invention has sarcosine, which is an amino acid, as a basic component and is excellent in biodegradability. In addition, a drug carrier containing the polysarcosine derivative of the present invention as one of the membrane components is excellent in biodegradability and excellent in blood retention (blood stability). Because of these characteristics, the drug carrier containing the novel polysarcosine derivative of the present invention as one of the membrane constituents and the pharmaceutical composition containing the drug carrier have excellent effects in preventing, treating or diagnosing diseases. Have.

  Hereinafter, the polysarcosine derivative of the present invention and a drug carrier containing the polysarcosine derivative as one of the membrane components will be described in detail.

（但し、式（１）中、Ｒ^１は水素原子、アミノ基の保護基またはアミノ基に結合する官能基であり、好ましくは水素原子またはアセチル基である。
また、Ｒ^２およびＲ^３は、互いに独立に炭素数１０〜２５のアルキル基またはアルケニル基であり、好ましくは互いに独立に炭素数１２〜１８のアルキル基またはアルケニル基である。
lは５〜１５０の整数を示し、好ましくは、１０〜１００の整数を示し、さらに好ましくは、２０〜８０の整数を示し、特に好ましくは、２０〜６０の整数を示す。） In the present invention, polysarcosine derivatives represented by the following general formulas (1) and (2) are provided.

(In the formula (1), R ¹ is a hydrogen atom, an amino protecting group or a functional group bonded to an amino group, preferably a hydrogen atom or an acetyl group.
R ² and R ³ are each independently an alkyl group or alkenyl group having 10 to 25 carbon atoms, and preferably are independently an alkyl group or alkenyl group having 12 to 18 carbon atoms.
l represents an integer of 5 to 150, preferably an integer of 10 to 100, more preferably an integer of 20 to 80, and particularly preferably an integer of 20 to 60. )

（但し、式（２）中、Ｒ^４は水素原子、アミノ基の保護基またはアミノ基に結合する官能基であり、好ましくは水素原子またはアセチル基である。
Ｒ^５およびＲ^６は互いに独立に炭素数１０〜２５のアルキル基またはアルケニル基であり、好ましくは互いに独立に炭素数１２〜１８のアルキル基またはアルケニル基である。
ｋは１０〜１００の整数を示し、好ましくは２０〜８０の整数を示し、更に好ましくは、２０〜６０の整数を示す。
ｍおよびｎは互いに独立に０または１〜３の整数を示し、好ましくは、ｍは２、ｎは０を示す。）
なお、本明細書において「アミノ基の保護基」とは、アセチル基、トリフルオロアセチル基、ベンジルオキシカルボニル基、ｔｅｒｔ-ブトキシカルボニル基などが含まれるが、これらに限定されるものではない。

(In the formula (2), R ⁴ is a hydrogen atom, an amino protecting group or a functional group bonded to an amino group, preferably a hydrogen atom or an acetyl group.
R ⁵ and R ⁶ are each independently an alkyl group or alkenyl group having 10 to 25 carbon atoms, preferably independently an alkyl group or alkenyl group having 12 to 18 carbon atoms.
k represents an integer of 10 to 100, preferably an integer of 20 to 80, and more preferably an integer of 20 to 60.
m and n each independently represent an integer of 0 or 1 to 3, preferably m is 2, and n is 0. )
In the present specification, the “amino-protecting group” includes, but is not limited to, an acetyl group, a trifluoroacetyl group, a benzyloxycarbonyl group, a tert-butoxycarbonyl group, and the like.

Accordingly, in the polysarcosine derivative represented by the general formula (1), R ¹ is preferably a hydrogen atom or an acetyl group, and R ² and R ³ are each independently an alkyl group or alkenyl group having 12 to 18 carbon atoms. It is preferable that L is preferably 20-60.
Specific examples of such a polysarcosine derivative include polysarcosine derivatives represented by the following structural formula (R ¹ is a hydrogen atom, R ² and R ³ are each independently an alkyl group having 18 carbon atoms, and l is 57). It is done.

In the polysarcosine derivative represented by the general formula (2), R ⁴ is preferably a hydrogen atom or an acetyl group, and R ⁵ and R ⁶ are each independently an alkyl group or alkenyl having 12 to 18 carbon atoms. It is preferably a group, m is 2, and n is preferably 0. Moreover, it is preferable that k is 20-60.
Specific examples of such a polysarcosine derivative include a polysarcosine derivative represented by the following structural formula (R ¹ is a hydrogen atom, R ² and R ³ are each independently an alkyl group having 18 carbon atoms, k is 23 or 48, m is 2 and n is 0).

（但し、式（３）中、Ｒ^２およびＲ^３は互いに独立に炭素数１０〜２５のアルキル基またはアルケニル基を示す） Hereinafter, the production method and purification method of the polysarcosine derivative represented by the above general formula (1) or (2) according to the present invention will be specifically described, but the production method and purification method of the present invention are limited to these. is not.
The polysarcosine derivative represented by the above general formula (1) is produced, for example, by polymerizing a compound represented by the following general formula (4) on the amino group of the compound represented by the following general formula (3). Can do.
In addition, the compound represented by the following general formula (4) is Y. Imanishi “N-Carboxyanhydres”, K.M. J. et al. Ivin and T.W. Saegusa "Ring-Opening Polymerization" Vol. 2, Elsevier, Essex, 1984, pp. 523-602 and references described therein.

(In the formula (3), R ² and R ³ independently represent an alkyl group or alkenyl group having 10 to 25 carbon atoms)

（但し、式（５）中、Ｒ^５およびＲ^６は互いに独立に炭素数１０〜２５のアルキル基またはアルケニル基である。ｍおよびｎは互いに独立に０または１〜３の整数を示す） The polysarcosine derivative represented by the general formula (2) can be produced, for example, by polymerizing the compound represented by the general formula (4) on the amino group of the compound represented by the following general formula (5). it can.

(In the formula (5), R ⁵ and R ⁶ are each independently an alkyl or alkenyl group having 10 to 25 carbon atoms. M and n each independently represents an integer of 0 or 1 to 3)

The amino compound represented by the general formula (5) can be synthesized by a general method, and specifically, a synthesis method referred to [Chemical Formula 12] can be given as an example.
The amino compound represented by the general formula (5) is obtained by protecting the amino group of a carboxylic acid derivative having an amino group (general formula (6)) with an appropriate protecting group (see “Protective Groups in Organic Synthesis” 3rd edition). , Theodora W. Greene, Peter GM Wuts, John Wiley & Sons Inc.), a carboxylic acid moiety having an amino group (general formula (7)) and an alkylamine derivative (general formula (8)). ) Can be synthesized by synthesizing a carboxylic acid derivative (general formula (9)) having an amino group in which an amide bond is formed by the condensation reaction, and then removing the amino group.

Examples of the carboxylic acid activator used in the reaction of the carboxylic acid derivative having an amino group (general formula (7)) and the alkylamine derivative (general formula (8)) include thionyl chloride, phosphorus pentachloride, Chloroformate (methyl chloroformate, ethyl chloroformate), oxalyl chloride, carbodiimides (eg, N, N′-dicyclohexylcarbodiimide (DCC), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (WSC)) Benzotriazol-1-yl-oxy-tris (dimethylamino) -phosphonium hexafluorophosphate (BOP) and the like. At this time, carbodiimides and N-hydroxybenzotriazole, 4-dimethylaminopyridine or hydroxysuccinimide may be used in combination.
In addition, the above reaction is usually performed by, for example, halogenated hydrocarbons such as methylene chloride and chloroform, ethers such as tetrahydrofuran (THF), dioxane, dimethyl ether, diethyl ether, isopropyl ether, N, N -It is carried out in the presence of dimethylformamide, N, N-dimethylacetamide or a mixed solvent thereof.
Moreover, the said reaction temperature is -10 degreeC-50 degreeC normally.

（但し、式（６）中、ｍおよびｎは互いに独立に０または１〜３の整数を示す。式（７）中、ｍおよびｎは互いに独立に０または１〜３の整数であり、Ｐは保護基を示す。式（９）中、Ｒ^５およびＲ^６は互いに独立に炭素数１０〜２５のアルキル基またはアルケニル基を示し、ｍおよびｎは互いに独立に０又は１〜３の整数であり、Ｐは、保護基を示す）

(In the formula (6), m and n each independently represent an integer of 0 or 1 to 3. In the formula (7), m and n are each independently an integer of 0 or 1 to 3; In formula (9), R ⁵ and R ⁶ each independently represent an alkyl group or alkenyl group having 10 to 25 carbon atoms, and m and n are each independently an integer of 0 or 1 to 3. And P represents a protecting group)

  The polysarcosine derivatives of the general formulas (1) and (2) synthesized by the above procedure can be isolated and collected by ordinary purification means such as chromatography and recrystallization.

The polysarcosine derivatives of the above general formulas (1) and (2) according to the present invention have sarcosine, which is an amino acid as a basic component, and are excellent in biodegradability. Disassembled. Since the sarcosine is a harmless substance in vivo, the polysarcosine derivatives of the above general formulas (1) and (2) according to the present invention can be used in vivo.
In the present specification, “biodegradable” means that a drug carrier containing the polysarcosine derivative of the above general formula (1) and / or (2) according to the present invention is administered to an administration subject (host) described later. , Refers to being degraded in the administration subject (host).

The drug carrier of the present invention is preferably a small spherical structure which is formed of a membrane having lipid as a basic constituent material and can carry a drug, and the constituent component (membrane constituent component) of the membrane is at least The polysarcosine derivative of the above general formula (1) and / or (2) is contained.
In addition, since various forms can be considered as the drug carrier of the present invention, in this specification, “support” widely means inclusion, encapsulation, adhesion by interaction, and the like depending on the form.

The form of the drug carrier of the present invention is not particularly limited as long as it has a structure capable of carrying the drug. For example, the form in which the drug can be enclosed in a closed space formed of a film, Examples include a form that can be encapsulated in the membrane itself, a form that allows the drug to adhere to the surface of the drug carrier, and a combination thereof.
Specific examples include liposomes having a structure capable of encapsulating a drug at a high concentration and closed vesicles such as lipid microspheres. Among them, the drug carrier of the present invention includes blood From the viewpoint of stability (retention in blood), liposomes are particularly preferable.

Liposomes are closed vesicles composed of phospholipid-based lipid bilayers, specifically, a space separated from the outside by a membrane formed based on the polarity of hydrophobic and hydrophilic groups of lipid molecules. And closed vesicles having a structure forming
A lipid microsphere is a closed vesicle formed by emulsifying one of water and oil in the other.

The size of the drug carrier of the present invention is not particularly limited, but when it takes a spherical shape or a form close thereto, the outer diameter of the particle is 0.02-250 μm, preferably 0.03-0.4 μm, more preferably Is 0.05 to 0.2 μm.
In the present specification, the “diameter of the particle outer diameter” refers to the average value of the diameters of all the drug carrier particles measured by the light scattering method.

The membrane component of the drug carrier of the present invention can contain only the polysarcosine derivative of the above formula (1) and / or (2) as a membrane component, but can stably form the structure of the drug carrier If so, other components may be contained together with the polysarcosine derivative of the above formula (1) and / or (2).
The “other component” is not particularly limited as long as it can stably form the structure of the drug carrier, but in view of its safety, in vivo stability, etc., it can be used as a membrane constituent component. And a derivative thereof, and among them, it is particularly preferable to include a lipid that is usually used as a basic constituent material of a membrane. Examples of lipids commonly used as the basic constituent material of the membrane include phospholipids, lipids other than phospholipids, cholesterols, and derivatives thereof.

Examples of the “phospholipid” include phosphatidylcholine (= lecithin), phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, cardioribine and the like, Synthetic phospholipids, or those hydrogenated according to these conventional methods (for example, hydrogenated soybean phosphatidylcholine) can be used.
In the present specification, “phospholipid” includes phospholipid derivatives obtained by modifying phospholipids.

Examples of the above-mentioned “lipid other than phospholipid” include lipids that do not contain phosphoric acid, and are not particularly limited, but glycerolipids that do not have a phosphate moiety in the molecule and sphingolipids that do not have a phosphate moiety in the molecule Can be mentioned.
In the present specification, “lipid other than phospholipid” includes derivatives of lipid other than phospholipid obtained by modifying lipid other than phospholipid.

When the lipid other than the phospholipid contains a basic functional group, that is, when the lipid is a substance in which a compound having a basic functional group is bound, the lipid is called a cationized lipid.
For example, when the drug carrier is a liposome, the cationized lipid is stabilized by including a part or all of the lipid portion as one of the components in the lipid bilayer membrane of the liposome. In addition, it is possible to modify the membrane of the drug carrier (liposome) by allowing the basic functional group moiety to exist on the membrane surface of the lipid bilayer (on the outer surface and / or the inner surface of the carrier). As a result, it is possible to enhance the adhesiveness with the cell that is the target site.

The “cholesterols” are not particularly limited, and examples thereof include cholesterol and cholestanol.
In addition, the said other component can be contained with the poly sarcosine derivative of said Formula (1) and / or (2) as 1 or a combination of 2 or more.

The content of the polysarcosine derivative of the formula (1) and / or (2) in the drug carrier of the present invention is a ratio to the total lipid amount constituting the drug carrier, and can usually be present at 0.1 to 20 mol%. , Preferably 0.1 to 5 mol%, more preferably 0.5 to 5 mol%.
If it is the said range, the adsorption | suction of a plasma protein, an opsonin protein, etc. can be prevented, the blood stability (retention property in blood) can be improved, and the effect which avoids the capture | acquisition by RES can be acquired.
In the present specification, the “total lipid amount” means the amount of all lipids constituting the drug carrier, and is expressed in molar concentration (mM). The “total lipid” means a lipid that is a membrane constituent of the drug carrier excluding the polysarcosine derivative of the formula (1) and / or (2). That is, specifically speaking, the total lipid means phospholipids, lipids other than phospholipids, or lipids such as cholesterols and their derivatives that form the basic membrane constituent material as described above.

The drug carrier of the present invention may contain a stabilizer and / or an antioxidant as the “other component” as long as the structure of the drug carrier can be stably formed.
Stabilizers include sterols such as cholesterol that reduce membrane fluidity, or sugars such as glycerol and sucrose.
Antioxidants include, but are not limited to, ascorbic acid, uric acid or tocopherol analogues such as vitamin E. Tocopherol has four isomers, α, β, γ, and δ, and any of them can be used in the present invention.
The stabilizer and the antioxidant are appropriately selected from the above as necessary, and are preferably used alone or in combination of two or more.

The drug carrier of the present invention can appropriately carry a drug for diagnosing, preventing or treating a disease in an amount within a range not impairing the effects of the present invention.
The kind of preventive or therapeutic drug that can be carried on the drug carrier of the present invention is not particularly limited as long as the structure of the drug carrier is not impaired. Specifically, nucleic acids, polynucleotides, genes and vectors containing them, anticancer agents, antibiotics, enzyme agents, antioxidant agents, lipid uptake inhibitors, hormone agents, anti-inflammatory agents, steroid agents, vasodilators, angiotensin conversion Enzyme inhibitor, angiotensin receptor antagonist, smooth muscle cell proliferation / migration inhibitor, platelet aggregation inhibitor, anticoagulant, chemical mediator release inhibitor, vascular endothelial cell proliferation promoter or inhibitor, aldose reductase inhibitor Agents, mesangial cell growth inhibitors, lipoxygenase inhibitors, immunosuppressive agents, immunostimulators, antiviral agents, amyloidosis inhibitors, nitric oxide synthesis inhibitors, AGEs (Advanced glycation endproducts) inhibitors, radical scavengers, proteins, peptides , Glycosaminogli Emissions and its derivatives, oligosaccharides and / or polysaccharides and derivatives thereof.

  Examples of the anticancer agent include, but are not limited to, nedaplatin, docetaxel, gemcitabine hydrochloride, cyclophosphamide, ifosfamide, nitrogen mustard-N-oxide, thiotepa, brusphan, carbocon, nimustine hydrochloride, ranimustine, melphalan, and improsul tosylate Fan, dacarbazine, procarbazine hydrochloride, cytarabine, cytarabine oxfate, enositabine, mercaptopurine, thioinosine, fluorouracil, doxyfluridine, tegafur, methotrexate, carmofur, hydroxycarbamide, vincristine sulfate, vinblastine sulfate, vindesine sulfate, etoposide hydrochloride, chromodamin A , Doxorubicin hydrochloride, araclubicin hydrochloride, pirarubicin, epirubicin hydrochloride Dactinomycin, mitoxantrone hydrochloride, bleomycin hydrochloride, peomycin, mitomycin C, neocarnostatin, L-asparaginase, acegraton mitopronitol, sodium dextran sulfate, octreotide acetate, cisplatin, carboplatin, tamoxifen citrate, medacetate Examples include roxyprogesterone, estramustine phosphate sodium, goserelin acetate, leuprorelin acetate, irinotecan hydrochloride, and paclitaxel. Among these anticancer agents, paclitaxel, docetaxel, irinotecan hydrochloride, gemcitabine hydrochloride, cisplatin, carboplatin, nedaplatin and the like are particularly preferable.

  Antibiotics include, but are not limited to, benzylpenicillin potassium, benzylpenicillin benzathine, phenoxymethylpenicillin potassium, pheneticillin potassium, cloxacillin sodium, flucloxacillin sodium, ampicillin, sultamicillin tosylate, bacampicillin hydrochloride, tarampicillin hydrochloride , Lenampicillin, hetacillin potassium, cyclacillin, amoxicillin, pibmesililin hydrochloride, aspoxicillin, carbenicillin sodium, calindacillin sodium, sulbenicillin sodium, ticarcillin sodium, piperacillin sodium, cephaloridine, cephalothin sodium, cefazolin sodium, cefapirin sodium, cefradine sodium , Cephalexin, cephatridin propylene glycol , Cefloxazine, cefaclor, cefadroxyl, cefothiam hydrochloride, cefotiam hexetyl hydrochloride, cefuroxime sodium, cefuroxime axetil, cefamandol sodium, cefdinir, cephemetomepipoxil hydrochloride, cefttipene, cefmetazole sodium, cefoxitin sodium , Cefotetan sodium, cefminox sodium, cefuperazone sodium, cefpyramide sodium, cefthrodine sodium, cefotaxime sodium, cefoperazone sodium, ceftizoxime sodium, cefmenoxime hydrochloride, ceftriaxone sodium, ceftazidime, cefpi Mizole sodium, cefixime, cefterampipoxil, cefzonam sodium, cefpodoxyproxetil, cefodizime, Cefpirom acid, latamoxef sodium, flomoxef sodium, imipenem, silastatin sodium, aztreonam, carmonam sodium, streptomycin sulfate, kanamycin sulfate, fradiomycin sulfate, amikacin sulfate, gentamicin sulfate, paromomycin sulfate, pecanamicin sulfate, Lipostamycin sulfate, dibekacin sulfate, tobramycin, sisomycin sulfate, mylonomycin sulfate, astromycin sulfate, netilmycin sulfate, isepamicin sulfate, arbekacin sulfate, erythromycin, kitasamycin, acetylkitasamycin, oleandomycin phosphate, josamycin, acetylspira Mycin, midecamycin, midecamycin acetate, rokitamycin, roxithromycin, clarithromycin, Tetracycline hydrochloride, oxytetracycline hydrochloride, tetracycline metaphosphate, demethylchlortetracycline hydrochloride, lolitetracycline, doxycycline hydrochloride, minocycline hydrochloride, chloramphenicol, chloramphenicol sodium succinate, chloramphenicol palmitate, thianphenicol, Aminoacetic acid thianphenicol hydrochloride, colistin sulfate, colistin sodium methanesulfonate, polymyxin B sulfate, bacitracin, vancomycin hydrochloride, lincomycin hydrochloride, clindamycin, spectinomycin hydrochloride, fosfomycin sodium, fosfomycin calcium and the like.

  Examples of the enzyme agent include, but are not limited to, chymotrypsin, crystalline trypsin, streptokinase / streptodolase, hyaluronidase, urokinase, nasarplase, alteplase, lysozyme chloride, semi-alkaline proteinase, serrapeptase, tisokinase, duteprase, batroxobin, pronase, promeline, etc. Can be mentioned.

  Antioxidants include, but are not limited to, tocopherol, ascorbic acid, uric acid and the like.

  Anti-inflammatory agents include, but are not limited to, choline salicylate, sazapyrine, sodium salicylate, aspirin, diflunisal, flufenamic acid, mefenamic acid, fructaphenine, tolfenamic acid, diclofenacnatrim, tolmetine sodium, sulindac, fenbufen, ferbinacethyl, indomethacin, indomethacin Farnesyl, acemetacin, progourmetacin maleate, ampenac sodium, nabumetone, ibuprofen, flurbiprofen, flurbiprofen axetil, ketoprofen, naproxen, protidic acid, pranoprofen, fenoprofen calcium, thiaprofenic acid, oxaprozin , Loxoprofen sodium, aluminoprofen, zaltoprofen, phenylbutazone Clofezone, keto phenylbutazone, piroxicam, tenoxicam, ampiroxicam, tiaramide hydrochloride, hydrochloric tinoridine, benzydamine hydrochloride, epirizole, Erumofazon the like.

  Steroid agents include, but are not limited to, cortisone acetate, hydrocortisone (phosphate ester, acetate), hydrocortisone butyrate, hydrocortisone sodium succinate, prednisolone (acetate, succinate, tertiary butyl acetate, phosphate ester), methylprednisolone (Acetate), methylprednisolone sodium succinate, triamcinolone, triamcinolone acetonide (triamcinolone acetate), dexamethasone (phosphate ester, acetate salt, sodium phosphate salt, sulfate ester), dexamethasone palmitate, betamethasone (phosphate, disodium) Salt), acetic acid parameterzone, fludrocortisone acetate, halopredone acetate, clobetasol propionate, halcinonide, beclomethasone propionate, Kusasan betamethasone, betamethasone acetate, acetic acid cortisone and the like.

  Examples of vasodilators include, but are not limited to, theophylline, diprofylline, proxyphylline, aminophylline, choline theophylline, prostaglandin, prostaglandin derivatives, alprostadil alphadex, alprostadil, limaprost alphadex, papaverine, cyclandelate, cinnarizine. , Bencyclane fumarate, cinepazide maleate, dirazep hydrochloride, trapidil, diphenidol hydrochloride, nicotinic acid, inositol hexanicotinate, nicotinate citrate, nicotinic alcohol tartrate, tocopherol nicotinate, hepronicart, isoxpurine hydrochloride, bamethane sulfate, tralizone hydrochloride, mesyl Dihydroergotoxin acid, ifenprodil tartrate, moxysilito hydrochloride, nicergoline, nicaric hydrochloride Pin, nilvadipine, nifedipine, benidipine hydrochloride, diltiazem hydrochloride, nisoldipine, nitrendipine, manidipine hydrochloride, balnidipine hydrochloride, efonidipine hydrochloride, verapamil hydrochloride, trimetazidine hydrochloride, captopril, enalapril maleate, alacepril, delapril hydrochloride, cilazapril hydrochloride, lisinapril hydrochloride, Examples include hydralazine, todralazine hydrochloride, budralazine, cadralazine, indapamide, carbochromen hydrochloride, efloxate, etaphenone hydrochloride, oxyfedrine hydrochloride, nicorandil, amyl nitrite, and isosorbide nitrate.

Examples of the angiotensin converting enzyme inhibitor include, but are not limited to, alacepril, imidapril hydrochloride, temocapril hydrochloride, delapril hydrochloride, benazepril hydrochloride, captopril, cilazapril, enalapril maleate, and lisinopril.
Examples of the angiotensin receptor antagonist include, but are not limited to, losartan.

  Smooth muscle cell proliferation and / or migration inhibitors include, but are not limited to, heparin sodium, dalteparin sodium (low molecular heparin), heparin calcium, dextran sulfate, and the like.

  Examples of the platelet aggregation inhibitor include, but are not limited to, ticlopidine hydrochloride, cilostazol, ethyl icosapentate, beraprost sodium, sarpgrelate hydrochloride, batroxobin, dipyridamole and the like.

  Examples of the anticoagulant include, but are not limited to, heparin sodium, dalteparin sodium (low molecular heparin), heparin calcium, dextran sulfate, warfarin potassium, argatroban and the like.

  Examples of chemical mediator release inhibitors include, but are not limited to, tranilast, ketofetin fumarate, azelastine hydrochloride, oxatomide, amlexanox, and repirinast.

  Although it does not specifically limit as an immunosuppressant, Cyclosporine etc. are mentioned.

  Antiviral agents include, but are not limited to, acyclovir, ganciclovir, didanosine, zidovudine, sorivudine, vidarabine and the like.

The kind of the diagnostic drug that can be carried on the drug carrier of the present invention is not particularly limited as long as the structure formation of the drug carrier is not impaired. Specific examples include in-vivo diagnostic agents such as X-ray contrast agents, ultrasonic diagnostic agents, radioisotope-labeled nuclear medicine diagnostic agents, and diagnostic agents for nuclear magnetic resonance diagnosis.
Examples of the X-ray contrast agent include meglumine amidotrizoate, sodium iotalamate, meglumine iotaramate, gastrografin, iodamid meglumine, lipiodol ultrafluid, adipiodon meglumine, oxaglic acid, meglumine iotroxate, iotrolane, iopanoic acid, iopamidol, Examples include iohexol, ioversol, iopodate sodium, iomeprol, isopaque, iodoxamic acid and the like.
Although it does not specifically limit as an ultrasonic diagnostic agent, For example, gas and a liquid are mentioned. Examples of the gas include air, carbon dioxide, oxygen, nitrogen, helium, and argon. Examples of the liquid include water, physiological saline, a buffer solution, and an aqueous suspension containing metal powder.

When the drug carrier of the present invention carries the drug, the size of the drug carrier of the present invention is not particularly limited. However, when the drug carrier has a spherical shape or a shape close thereto, the diameter of the particle outer diameter is 0. It is 02 to 250 μm, preferably 0.03 to 0.4 μm, more preferably 0.05 to 0.2 μm.
In particular, in the range of 0.05 to 0.2 μm, an EPR effect (Enhanced Permeability and Retention Effect) can be expected.
The EPR effect is an effect that the drug carrier of the present invention tends to stay in the tumor or inflamed tissue of the administration subject (host). Blood vessels in tumor tissues and inflamed tissues are more permeable than blood vessels in normal tissues. Therefore, blood vessels in tissues such as tumors are 0.05 to 0.2 μm in size compared to blood vessels in normal tissues. Molecular compounds are easy to permeate. In such a tumor tissue or the like, since the lymphatic mechanism is incomplete, the drug carrier or the like transferred to the tissue stays in the tissue without being collected from the lymphatic vessel. Such an effect is called EPR.

The drug carrier of the present invention can be produced, separated and purified by employing known methods. Hereinafter, as an example of the drug carrier, a liposome production method and a separation / purification method will be described, but the drug carrier production method and the separation / purification method of the present invention are not limited thereto.
As a method for producing, separating and purifying the drug carrier of the present invention, first, in a flask, the polysarcosine derivative according to the general formula (1) and / or (2) according to the present invention and other membranes such as phospholipids are used. The constituent component is mixed with an organic solvent such as chloroform, and the organic solvent is distilled off, followed by vacuum drying to form a thin film on the inner wall of the flask. Next, a liposome dispersion is obtained by vigorous stirring. In addition, when carrying | supporting a chemical | medical agent, it stirs, after adding the chemical | medical agent carry | supported in the said flask.
The obtained liposome dispersion can be purified by any commonly used method such as gel filtration, dialysis, membrane separation, and / or centrifugation to remove the drug not supported on the liposome. Moreover, the obtained liposome dispersion liquid can adjust the outer diameter of a liposome particle by methods used normally, such as a French press, a pressure filter, or an extruder.
In addition, after forming the liposome by a conventional method using the mixed lipid obtained by mixing the liposome-forming lipid not containing the polysarcosine derivative described in the general formula (1) and / or (2) according to the present invention, You may add the poly sarcosine derivative as described in General formula (1) and / or (2) by this invention.

  In addition to the above method, the liposome can also be obtained by mixing the above-described constituent components and discharging them with a high-pressure discharge type emulsifier. The method for preparing liposomes is specifically described in “Liposomes in Life Science” (Terada, Yoshimura et al .; Springer Fairlark Tokyo (1992)), and is described in this specification with reference to this description. Shall.

A pH gradient method can be used to support the drug on the drug carrier of the present invention. This method is described in “Liposome Technology Liposome Preparation and Related Technologies 2nd Edition” (Edited by G. Gregoriadis, Vol. I-III, CRC Press), and is described in this specification with reference to this description. It shall be.
Several techniques are available for sizing the liposomes of the present invention to a desired size, but as an example, “Liposome Technology Liposome Preparation and Related Technologies 2nd Edition” (Edited by G. Gregoriadis, Vol. I- III, CRC Press). It is assumed that the present description is described with reference to this description.

The drug carrier of the present invention suppresses aggregation due to interaction with opsonin protein and plasma protein in blood, capture in reticuloendothelial tissue (RES) such as liver and spleen, etc., and retention in blood Since the sex (blood stability) can be increased, the drug can be transported to the target site while being supported, and as a result, the supported drug can be delivered to the target site.
In the present specification, “high retention in blood (blood stability)” means that after administration of a drug carrier to an administration target (host) of a drug carrier described later, The state which exists for a longer time compared with the drug carrier which does not contain the polysarcosine derivative as described in the general formula (1) and / or (2) according to the invention.
In the present specification, the “target site” is not particularly limited as long as it is a site to be prevented, treated or diagnosed. Specific examples include internal structures such as tumors, inflamed cells, tissues, organs (eg, organs), and cell nuclei.
In the present specification, “transportation of a drug to a target site” means that the drug carrier carrying the drug is transported from the point where it is administered to the host (administration target) to the target site in the living body.
Specifically, the drug carried on the drug carrier of the present invention may be transported to a target site in the living body by utilizing the EPR effect (Enhanced Permeation and Retention effect).
In the present specification, “delivery of a drug to a target site” means that a drug carried by a drug carrier reaches and takes up the target site. In this case, it also includes the effect of the drug on the target site or its vicinity even if the drug is not taken into the target site.

  The pharmaceutical composition of the present invention comprises a composition in which a drug is carried on the drug carrier of the present invention, and additives such as the above-mentioned stabilizers and antioxidants are optionally contained as necessary.

In the pharmaceutical composition of the present invention, the drug to be carried on the drug carrier of the present invention is appropriately selected from the above preventive or therapeutic drugs as necessary, and used in combination of one or two or more. Preferably it is done.
In the pharmaceutical composition of the present invention, the drug carrier of the present invention can contain additives such as the above-mentioned stabilizers and antioxidants depending on the administration route and dosage form.
Examples of the additive include, but are not limited to, the stabilizer, the antioxidant, water, physiological saline, pharmaceutically acceptable organic solvent, collagen, polyvinyl alcohol, polyvinyl pyrrolidone, carboxyvinyl polymer, Sodium carboxymethylcellulose, sodium polyacrylate, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin, methylcellulose, ethylcellulose, xanthan gum, gum arabic, casein, gelatin, agar, diglycerin, propylene glycol, polyethylene glycol, petroleum jelly, paraffin , Stearyl alcohol, stearic acid, human serum albumin (HSA), mannitol, sorbitol, lactose, PBS, biodegradable polymer, bloodless Examples include a medium, a surfactant that is acceptable as a pharmaceutical additive, or a buffer solution of physiological pH that is acceptable in vivo. From these, one or two or more is appropriately selected as necessary. Preferably used in combination.
The pharmaceutical composition of the present invention can be stored according to an ordinary method, for example, refrigerated at 0 to 8 ° C or stored at a room temperature of 15 to 25 ° C.

  The disease or pathological condition to which the pharmaceutical composition of the present invention is applied, or a specific therapeutic index in them, includes various types and is not necessarily limited. Any disease can be applied to which the various drugs described above can be applied.

The administration target (host) of the pharmaceutical composition of the present invention includes mammals, preferably humans, monkeys, mice, livestock, and the like.
The dosage of the pharmaceutical composition of the present invention is not particularly limited and is appropriately determined in consideration of the symptoms, body weight, age, sex, etc. of the administration subject (host), but the administration subject (host) already suffering from the disease. In addition, it is desirable to administer in an amount effective to cure or at least partially block the symptoms of the disease. In addition, it is desirable to administer in an effective amount that can be used to determine the diagnosis of the disease.
For example, when the administration subject (host) is a human, the effective dose of the drug carried on the drug carrier is preferably selected in the range of 0.01 mg to 100 mg per kg body weight per day.
The administration time may be administered after the disease has occurred, or may be administered prophylactically to relieve symptoms at the time of onset when the onset of the disease is predicted. The administration period can be appropriately selected depending on the age, symptoms, etc. of the administration subject (host).

Although the dosage form of the pharmaceutical composition of this invention is not necessarily limited, an injection etc. are mentioned. Various administration routes are possible, but parenteral administration is preferred. As parenteral administration, for example, injection such as intravenous administration (continuous infusion, intermittent infusion), intraperitoneal administration, subcutaneous administration, intramuscular administration, intraarterial administration and the like is common.
As a specific administration method, the pharmaceutical composition of the present invention is administered by syringe or infusion, a catheter is inserted into the body of a patient or host, for example, into a lumen, for example, into a blood vessel, and the tip thereof is near the target site. And a method of administering through the catheter from a desired target site, the vicinity thereof or a site where blood flow to the target site is expected.
Moreover, although the administration time or frequency | count also depends on a patient's condition, preventive administration, single administration, continuous administration, etc. are illustrated.

  The pharmaceutical composition of the present invention can be obtained by the same method as the production and separation method described in the drug carrier of the present invention.

  Since the pharmaceutical composition of the present invention is a composition containing the drug carrier of the present invention, it can be transported to the target site in a state where the drug is loaded, and as a result, the loaded drug is delivered to the target site. can do. Therefore, it has an excellent effect in preventing, treating or diagnosing diseases.

  The present invention provides a preventive method, a therapeutic method or a diagnostic method using a pharmaceutical composition comprising the drug carrier of the present invention as an active ingredient for a disease or condition to which the above-described pharmaceutical composition of the present invention is applied. it can.

  EXAMPLES Hereinafter, although this invention is demonstrated in more detail based on an Example and a test example, this invention should not be limited to these Examples and a test example.

(Example _{1) (Sar) 57 -DODA (} ( sarcosine) ₅₇ - di octadecanyl amine) Synthesis of (1) Sar NCA synthesized Sar NCA using synthetic thionyl chloride method (sarcosine N- carboxyanhydride) did. Thionyl chloride was added to carbobenzyloxy sarcosine (Z-Sar) and reacted at 60 ° C. for about 7 minutes. The reaction solution was added to dry petroleum ether, and the resulting precipitate was collected. The solid obtained was recrystallized three times using a mixed solvent of ethyl acetate and petroleum ether, and purification of the target product (Sar NCA) was confirmed by melting point measurement (melting point 97 ° C.).
(2) Polymerization Sar NCA was dissolved in tetrahydrofuran, and an amount corresponding to the degree of polymerization, n = 60, with DODA set under stirring at 50 ° C. was added. The monomer concentration was set to 0.2M. After the reaction for 4 hours, disappearance of the NCA characteristic absorption peak was confirmed by IR spectrum measurement. The reaction solution was concentrated under reduced pressure and dried under reduced pressure to obtain the target compound ((Sar) ₅₇ -DODA).
The ¹ H NMR spectrum of the obtained compound ((Sar) ₅₇ -DODA) is shown in FIG.

The ¹ H NMR spectrum in FIG. 1 supports the structure of the following formula.

(Example 2) Synthesis of Ac- (Sar) ₄₀ -DODA Sar NCA synthesized in (1) of Example 1 was dissolved in tetrahydrofuran, and the degree of polymerization was set at 50 ° C. with stirring and DODA was set to n = 40. The corresponding amount was added. The monomer concentration was set to 0.2M. After the reaction for 4 hours, disappearance of the NCA characteristic absorption peak was confirmed by IR spectrum measurement. Acetic acid (5 equivalents), HATU (N-[(dimethylamino) -1H-1,2,3-triazol [4,5-b] pyridino-1-ylmethylene] -N-methylmethanamine hexafluorophosphate N-oxide) were added to the reaction solution. 5 equivalents) and diisopropylpropylamine (DIEA) (7.5 equivalents) were added to acetylate the N-terminus. After the reaction for 12 hours, HATU (2 equivalents) was added. After a further 12 hours of reaction, the reaction solution was concentrated under reduced pressure, and the resulting solid was dissolved in methanol and purified using a Sephadex ^™ LH20 column (2 cm × 65 cm eluent: methanol). FIG. 2 shows the elution pattern of Ac-Sar ₄₀ -DODA. A fraction (fraction number: 19 to 24) having an elution volume of 57 to 72 ml was collected, concentrated under reduced pressure, and dried under reduced pressure to obtain the target compound. The ¹ H NMR spectrum of the obtained compound is shown in FIG.

The ¹ H NMR spectrum of [FIG. 3] supports the structure of the following formula.

(Example 3) Synthesis of Ac- (Sar) _24- DODA Sar NCA synthesized in (1) of Example 1 was dissolved in tetrahydrofuran, and the degree of polymerization was set to DO = 20 at 50 ° C with stirring. The corresponding amount was added. Thereafter, the target compound was obtained in the same manner as in (Example 2). The ¹ H NMR spectrum of the obtained compound (Ac- (Sar) ₂₄ -DODA) is shown in FIG.

The ¹ H NMR spectrum of FIG. 4 supports the structure of the following formula:

Example 4 Synthesis of (Sar) ₂₃ -Glu (ODA) ₂ (1) Synthesis of amine derivative Boc-Glu (2.5 g), N-hydroxysuccinimide (3.49 g), stearylamine (5. 98 g) was dissolved in dichloromethane, and dicyclohexyldiimide (4.99 g) was added with stirring at 0 ° C. After 3 hours, the temperature was returned to room temperature, and the mixture was allowed to react overnight with stirring. The precipitated solid was collected by filtration, washed with methanol, and the target product was dissolved in warm tetrahydrofuran. The solution was concentrated under reduced pressure, and chloroform was added to the precipitated solid to extract the desired product. Recrystallization was performed from a mixed solvent of chloroform and methanol. The ¹ H NMR spectrum of the obtained solid Boc protector (Boc-Glu (ODA) -ODA) is shown in FIG.

得られたＢｏｃ-Ｇｌｕ（ＯＤＡ）-ＯＤＡ（１．６ｇ）にトリフルオロ酢酸（１０ｍｌ）を加え、室温で３０分反応後、減圧濃縮した。得られた固体をクロロホルムに溶解し、４％ 炭酸水素ナトリウム水溶液（ＮａＨＣＯ_３ａｑ）で２回処理した。塩化ナトリウム水溶液（ＮａＣｌａｑ）で洗浄後、クロロホルム層を無水硫酸マグネシウムで乾燥した。減圧濃縮して得られた固体をＮ,Ｎ−ジメチルホルムアミドで洗浄後、テトラヒドロフランとメタノールの混合溶媒から再結晶し、開始剤 アミン体（ＮＨ_２-Ｇｌｕ（ＯＤＡ）-ＯＤＡ）を得た。なお、（ＮＨ_２-Ｇｌｕ（ＯＤＡ）-ＯＤＡ）は下記式の構造を支持する。

（２）重合
実施例１の（１）で合成したＳａｒ ＮＣＡをテトラヒドロフランに溶解し、５０℃で攪拌下にＮＨ_２-Ｇｌｕ（ＯＤＡ）-ＯＤＡを設定した重合度、ｎ=２０に相当する量を加えた。モノマー濃度は０．２Ｍに設定した。１晩反応後、反応溶液を減圧濃縮し、得られた固体をメタノールに溶解して、Ｓｅｐｈａｄｅｘ^ＴＭ ＬＨ２０カラムを用いて精製した。得られたポリマーの¹Ｈ ＮＭＲスペクトルを［図６］に示す。 The ¹ H NMR spectrum in FIG. 5 supports the structure of Boc-Glu (ODA) -ODA.

Trifluoroacetic acid (10 ml) was added to the obtained Boc-Glu (ODA) -ODA (1.6 g), reacted at room temperature for 30 minutes, and concentrated under reduced pressure. The obtained solid was dissolved in chloroform and treated twice with 4% aqueous sodium hydrogen carbonate solution (NaHCO ₃ aq). After washing with a sodium chloride aqueous solution (NaClaq), the chloroform layer was dried over anhydrous magnesium sulfate. The solid obtained by concentration under reduced pressure was washed with N, N-dimethylformamide and then recrystallized from a mixed solvent of tetrahydrofuran and methanol to obtain an initiator amine (NH ₂ -Glu (ODA) -ODA). Note that (NH ₂ -Glu (ODA) -ODA) supports the structure of the following formula.

(2) Polymerization Sar NCA synthesized in (1) of Example 1 was dissolved in tetrahydrofuran, and the degree of polymerization was set to NH ₂ -Glu (ODA) -ODA with stirring at 50 ° C., an amount corresponding to n = 20 Was added. The monomer concentration was set to 0.2M. After reacting overnight, the reaction solution was concentrated under reduced pressure, and the resulting solid was dissolved in methanol and purified using a Sephadex ^™ LH20 column. The ¹ H NMR spectrum of the obtained polymer is shown in FIG.

The ¹ H NMR spectrum of FIG. 6 supports the structure of the following formula:

Example 5 Synthesis of (Sar) ₄₈ -Glu (ODA) ₂ Sar NCA synthesized in (1) of Example 1 was dissolved in tetrahydrofuran and stirred at 50 ° C. under stirring with NH ₂ -Glu (ODA) -ODA. Was added in an amount corresponding to the degree of polymerization, n = 40. The monomer concentration was set to 0.2M. After reacting overnight, the reaction solution was concentrated under reduced pressure, and the resulting solid was dissolved in methanol and purified using a Sephadex ^™ LH20 column. The ¹ H NMR spectrum of the obtained polymer is shown in FIG.

The ¹ H NMR spectrum of FIG. 7 supports the structure of the following formula:

(Example 6)
Liposomes containing the polysarcosine derivative synthesized in Example 1 as one of the membrane constituents were prepared as shown in [Table 1] below.
Hydrogenated soybean phosphatidylcholine (HSPC, Lipoid) 0.312 mmol, cholesterol (Solvay) 0.266 mmol, polysarcosine derivative synthesized in Example 1 0.029 mmol (modification rate 5 mol%) was weighed, and ethanol was added in an amount of 0. 5ml was added and heated to dissolve the lipid, and then 4.5ml of physiological saline was added to the filter (pore size, 0.2μm × 5 times, 0.1μm × 10 times) attached to the extruder at 65 ° C. Through the liposomes were prepared. As a result, a liposome containing the compound synthesized in Example 1 as one of membrane constituents was obtained. The “modification rate” means the ratio of the polysarcosine derivative to the total lipid amount.

(Example 7)
Liposomes containing the polysarcosine derivative synthesized in Example 2 as one of the membrane constituents were prepared as shown in [Table 1] below.
Liposomes were prepared in the same manner as in Example 6 except that the polysarcosine derivative synthesized in Example 2 was used instead of the polysarcosine derivative synthesized in Example 1.

(Example 8)
Liposomes containing the polysarcosine derivative synthesized in Example 3 as one of the membrane constituents were prepared as shown in [Table 1] below.
Liposomes were prepared in the same manner as in Example 6 except that the polysarcosine derivative synthesized in Example 3 was used instead of the polysarcosine derivative synthesized in Example 1.

Example 9
Liposomes labeled with a fluorescent dye were prepared as shown in [Table 2] below.
Hydrogenated soybean phosphatidylcholine (HSPC, Lipoid) 0.312 mmol, cholesterol (Solvay) 0.266 mmol, polysarcosine derivative 0.029 mmol synthesized in Example 1 or 2 was weighed (0. 029 mmol, 2 mol%, 0.012 mmol in the case of modification), rhodamine-DHPE (Lissamine ^™ , rhodamine B 1,2-dihezadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt, Molecular Probes 0.5 ml of ethanol solution (3.09 mg / ml) / warm, and after lipids are dissolved, 4.5 ml of physiological saline is added and a filter (pore size) attached to an extruder at 65 ° C. , 0. [mu] m × 5 times, passed through a 0.1 [mu] m × 10 times), liposomes were prepared. As a result, liposomes containing the compound synthesized in Examples 1 to 3 as one of the membrane constituents were obtained.
The average particle size of each liposome of Example 9 is shown in [Table 3].

(Reference Example 1)
Preparation of liposomes labeled with a fluorescent dye containing polyethylene glycol (molecular weight 2000) -phosphatidylethanolamine (PEG-PE) as one of the membrane components Hydrogenated soybean phosphatidylcholine (HSPC, Lipoid) 0.312 mmol , 0.266 mmol of cholesterol (Solvay), 0.029 mmol of polyethylene glycol (molecular weight 2000) -phosphatidylethanolamine (PEG-PE) (PEG-PE modification rate 5 mol%), rhodamine-DHPE (fluorescent dye) Lissamine ^TM, Zadekanoiru to rhodamine B 1,2-di -sn- glycero 3-phosphotransferase, triethylammonium salt (Molecular Probes, Inc.) / ethanol solution (3.0 0.5 ml of mg / ml) was added and heated to dissolve the lipid, and then 4.5 mL of physiological saline was added, and the filter (pore size, 0.2 μm × 5 times, 0) attached to the extruder at 65 ° C. As a result, liposomes containing polyethylene glycol (molecular weight 2000) -phosphatidylethanolamine (PEG-PE) as one of the membrane constituents were obtained. The “PEG-PE modification rate” means the ratio of PEG-PE to the total lipid amount.
The average particle size of each liposome of Example 9 and Reference Example 1 is shown in [Table 3].

(Test Example 1)
Blood concentration after administration of each sample of the liposome of Example 9-1, the liposome of Example 9.2, the liposome of Example 9-3, and the liposome of Reference Example 1 using rats (male, 7 weeks old) Was observed with fluorescence of rhodamine DSPE as a label.
A sample corresponding to a total lipid amount of 10 μmol / 2 ml / kg was injected from the rat tail vein, and about 0.5 ml of blood was collected from the tail vein using a heparinized syringe 1, 3, 6, 24, and 48 hours after administration. Using 0.1 ml of blood, the fluorescence intensity of rhodamine in the blood was measured using a fluorometer (excitation 550 nm, fluorescence 590 nm). The total amount of lipid in blood (μmol total lipid / ml blood) was calculated from the obtained fluorescence intensity. The time course of the total lipid amount is shown in FIG.

  It was confirmed that 10% or more of the liposome of Example 9-1, the liposome of Example 9-2, and the liposome of Example 9-3 remained in the blood vessel even after 24 hours had passed from the blood vessel. The blood stability (retention in blood) was almost the same as that of the liposome. The long-term retention of the liposome of Example 9-1, the liposome of Example 9.2, and the liposome of Example 9-3 indicates that the drug can be accumulated in the cancer affected area by the EPR effect. Yes.

(Acute toxicity)
As a result of an acute toxicity test by oral administration using ICR male mice (5 weeks old), the LD50 of the compound of the present invention (polysarcosine derivative represented by the general formula (1) or (2)) is 320 mg. / Kg or more, and the safety of the drug carrier containing the polysarcosine derivative was confirmed.

FIG. 1 shows a ¹ H NMR spectrum of (Sar) ₅₇ -DODA. FIG. 2 is a diagram showing the elution pattern of Ac- (Sar) ₄₀ -DODA in CDCl ₃ by Sephadex ^™ LH20 chromatography. FIG. 3 shows a ¹ H NMR spectrum of Ac- (Sar) ₄₀ -DODA in CDCl ₃ . FIG. 4 shows a ¹ H NMR spectrum of Ac- (Sar) ₂₄ -DODA in CDCl ₃ . FIG. 5 is a diagram showing a ¹ H NMR spectrum of (Boc-Glu (ODA) -ODA) in CDCl ₃ . FIG. 6 is a diagram showing a ¹ H NMR spectrum of (Sar) ₂₃ -Glu (ODA) ₂ in CDCl ₃ . FIG. 7 shows the ¹ H NMR spectrum of (Sar) ₄₈ -Glu (ODA) ₂ in CDCl ₃ . FIG. 8 is a diagram showing the change over time of the liposome concentration in rat blood.

Claims (9)

A polysarcosine derivative represented by the following general formula (1).

(In the formula (1), R ¹ is a hydrogen atom, an amino-protecting group or a functional group bonded to an amino group, and R ² and R ³ are each independently an alkyl group or alkenyl having 10 to 25 carbon atoms. (L represents an integer of 5 to 150) A polysarcosine derivative represented by the following general formula (2).

(In the formula (2), R ⁴ is a hydrogen atom, an amino protecting group, or a functional group bonded to an amino group, and R ⁵ and R ⁶ are each independently an alkyl or alkenyl having 10 to 25 carbon atoms. (K represents an integer of 10 to 100. m and n each independently represents an integer of 0 or 1 to 3) ^{2. The} polysarcosine derivative according to claim 1, wherein in the general formula (1), R ² and R ³ are each independently an alkyl group or an alkenyl group having 12 to 18 carbon atoms. 3. The polysarcosine derivative according to claim 2, wherein, in the general formula (2), R ⁵ and R ⁶ are each independently an alkyl group or an alkenyl group having 12 to 18 carbon atoms. 4. The polysarcosine derivative according to claim 1, wherein, in the general formula (1), R ² and R ³ are an alkyl group having 18 carbon atoms, and 1 is 20 to 60. 5. 5. The polysarcosine derivative according to claim 2, wherein in the general formula (2), R ⁵ and R ⁶ are an alkyl group having 18 carbon atoms, and k is 20 to 60. 6.   A drug carrier comprising the polysarcosine derivative according to any one of claims 1 to 6 as one of constituent components.   8. The drug carrier according to claim 7, wherein the drug carrier is a closed vesicle.   The drug carrier according to claim 7 or 8, further comprising at least one selected from the group consisting of phospholipids, glycerolipids other than phospholipids, sphingolipids, and cholesterols.

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