KR100413532B1 - Polyamine terminated dendrimers and preparing method thereof - Google Patents

Abstract

The present invention provides a polyammonium dendrimer substituted with a polyamine group at its terminal, a method for preparing the same, and a use thereof. The present invention also provides a pseudorotaxane dendrimer derivative which is a self-assembled complex between a polyamine group and cucurbitryl at the end of the dendrimer, a method for preparing the same, and a use thereof. The polyammonium dendrimer and the pseudo-rotaxane dendrimer derivative may trap and trap a biochemically active substance inside the dendrimer, and slowly release the biochemically active substance trapped therein as the complex formed is dissociated. In addition, the polyammonium dendrimer may form a self-assembled complex through ion interaction with a biochemically active substance such as polynucleotide, DNA, or the like. Therefore, such materials can be usefully used in biochemical fields such as drug delivery, gene therapy, gene delivery, and the like.

Description

Polyamine terminated dendrimers and preparing method

The present invention relates to a dendrimer substituted with a polyamine at the end of the dendrimer, and more particularly, to a polyammonium dendrimer having a polyamine substituted at the end of the dendrimer, a method for producing the polyamine and a cooker at the end of the polyammonium dendrimer The present invention relates to a self-assembly complex formed by the reaction between the vitriles and a manufacturing method thereof.

Dendrimers are polymers with many branches in the form of three-dimensional spaces with well-organized chemical structures, consisting of a core region, a branch region, and an outer surface (terminal) region. The dendrimer has a shape in which an inner space is formed by a branching region. Thus, dendrimers can capture large molecules such as dyes and fluorescent materials and metal ions, as well as solvents and small organic materials, and can be used as carriers.

Currently, research has been conducted on dendrimer boxes that change the chemical environment of molecules trapped in the inner space of the dendrimer so that the dendrimer can come out of the dendrimer (US Pat. No. 5,990,089). This study is possible by changing the terminal region of the dendrimer.

On the other hand, the biggest characteristic of the dendrimer is that there are many functional groups in the terminal region. Therefore, chemically modifying functional groups present on the surface not only changes the properties of the dendrimer itself, but also changes the physical and mechanical properties, thereby synthesizing compounds having functionality. In addition, dendrimers have many of the same functional groups on their surfaces, and thus they can be used to obtain functional synergies ( J. Polym. Sci .: Part B: Polym. Phys. , 1995 , 33 , 333). That is, the hydrophilic surface of the dendrimer may be changed to lipophilic, and the nucleophilic dendrimer may be changed to electrophilic.

The technical problem to be achieved by the present invention is to provide a polyammonium dendrimer having a polyamine group in the terminal region, which is capable of self-assembling with a biochemically active material, a method for producing the same and a use thereof.

Another technical problem to be achieved by the present invention is to provide a self-assembly complex formed by the reaction between the polyamine at the dendrimer end and the cucurbitryl, which can be used as a collection and carrier of biochemically active materials as a dendrimer box, a method for producing the same, and a use thereof. will be.

In order to achieve the first technical problem, the present invention provides a polyammonium dendrimer represented by Chemical Formula 1.

Wherein R ₁ and R ₂ are each independently a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, and ○ is a poly (propyleneimine) dendrimer and a poly (amidoamine) dendrimer A dendrimer center region selected from the group consisting of: Y is O or NH, m is the number of functional groups on the surface according to the generation number of the dendrimer, an integer from 3 to 200, n is an integer from 1 to 3, x is Is an integer from 2 to 10 and X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate.

A second technical object of the present invention is to obtain an ester compound (B) by (a) reacting a diamine compound (A) with an alkyl bromoacetate and reacting it with an amine protecting group or with a protected aminoalkyltosylate;

(b) hydrolyzing the ester compound (B) to obtain a carboxylic acid compound (C);

(c) obtaining an ester compound (D) by esterification of the carboxylic acid compound (C);

(d) reacting a dendrimer (E) with one selected from the carboxylic acid compound (C) and the ester compound (D) to obtain a compound (F); And

(e) performing a deprotection reaction of the protected amine group of the compound (F); and by the method for producing a polyammonium dendrimer represented by the formula (1).

Wherein R ₁ and R ₂ are each independently a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, and R ₃ , R ₄ and R ₅ are each independently a hydrogen atom or carbon number Is selected from the group consisting of 2 to 4 protected aminoalkyl groups, protected diaminoalkyl groups of 5 to 10 carbon atoms, benzyloxycarbonyl groups, t-butoxycarbonyl groups and p-toluenesulfonyl groups, and R ₆ is 1 to 5 carbon atoms Is an alkyl group, R ₇ is a pentafluorophenyl group or a succinimide group, ○ represents a dendrimer center selected from the group consisting of poly (propyleneimine) dendrimers and poly (amidoamine) dendrimers, and Y is O or NH , m is the number of functional groups on the surface according to the generation number of the dendrimer, n is an integer from 3 to 200, n is an integer from 1 to 3, x is an integer from 2 to 10, X is a halogen atom, hexaflu In phosphate, it is selected from the group consisting of trifluoroacetate.

The third technical problem of the present invention is achieved by a pseudorotaxane dendrimer derivative represented by the formula (2).

Wherein R ₂ is a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, and ○ is a dendrimer selected from the group consisting of poly (propyleneimine) dendrimers and poly (amidoamine) dendrimers Represents a center, Y is an oxygen atom or an amino group, m is the number of functional groups on the surface according to the generation number of the dendrimer, an integer of 3 to 200, n is an integer of 1 to 3, and x is an integer of 2 to 10 And X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate.

The fourth technical problem of the present invention is achieved by a method for preparing a pseudorotaxane dendrimer derivative represented by Chemical Formula 2, wherein an excess of cucurbityl is added to the polyammonium dendrimer of Chemical Formula 1 to react.

<Formula 1>

<Formula 2>

Wherein R ₁ and R ₂ are each independently a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, and ○ is a poly (propyleneimine) dendrimer and a poly (amidoamine) dendrimer A dendrimer center region selected from the group consisting of: Y is O or NH, m is the number of functional groups on the surface according to the generation number of the dendrimer, an integer of 3 to 200, n is an integer of 1 to 3, x Is an integer from 2 to 10, and X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate.

The fifth technical problem of the present invention is achieved by a composition for capturing and delivering a biochemically active substance, comprising the polyammonium dendrimer or pseudorotaxane dendrimer derivative.

Hereinafter, the principle of the present invention will be described.

Dendrimers in which amine groups are introduced on the outer surface, such as poly (propyleneimine) dendrimers or poly (amidoamine) dendrimers, can be used for polynucleotide delivery systems and gene transfection. This is because the primary amine present on the dendrimer surface is changed to ammonium cation so that the dendrimer polycation can interact with non-chemically covalently with biochemically active materials such as polynucleotides, DNA, and the like.

Accordingly, the present inventors have completed the present invention regarding a polyammonium dendrimer in which primary ammonium ions are changed to polyammonium cations on the dendrimer surface in consideration of the synergistic effect of non-covalent bonds as described above. The polyammonium cation can easily form a complex, ie, rotacic acid, by self-assembly with cucurbitryl, which is a synthetic receptor, to make supramolecular dendrimers (rotaxane dendrimers). At this time, the cucurbitryl pseudorotaxanes formed on the dendrimer surface form a hard shell and prevent the release of the substances trapped inside the dendrimer. The dissociation of the cucurbitel of the pseudorotaxane on the surface dissipates the surface of the dendrimer and releases the substance trapped therein.

Polyammonium dendrimer according to the present invention is represented by the following formula (1).

<Formula 1>

Wherein R ₁ and R ₂ are each independently a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, and ○ is a poly (propyleneimine) dendrimer and a poly (amidoamine) dendrimer A dendrimer center region selected from the group consisting of: Y is O or NH, m is the number of functional groups on the surface according to the generation number of the dendrimer, an integer from 3 to 200, n is an integer from 1 to 3, x Is an integer from 2 to 10, and X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate.

In particular, the polyammonium dendrimer of Formula 1 is a compound wherein R ₁ and R ₂ are hydrogen atoms, n is 1 to 3, Y is O or NH, and X is Br, Cl, PF ₆ , wherein R ₁ is A hydrogen atom, R ₂ is an aminopropyl group, n is 1 to 3, Y is O or NH, X is Br, Cl, PF ₆ , R ₁ and R ₂ are aminopropyl groups, n Is 1 to 3, Y is O or NH, X is Br, Cl, PF ₆ or R ₁ is an aminopropyl group, R ₂ is a hydrogen atom, n is 1 to 3, and Y is O Or NH and X is Br, Cl, PF ₆ .

With reference to Schemes 1 and 2, it will be described how to prepare a polyammonium dendrimer of the above formula (1).

R ₃ , R ₄ and R ₅ independently of one another are a hydrogen atom, a protected aminoalkyl group having 2 to 4 carbon atoms, a protected diaminoalkyl group having 5 to 10 carbon atoms, benzyloxycarbonyl group, t-butoxycarbonyl group and p-toluenesul Is selected from the group consisting of

R ₆ is an alkyl group having 1 to 5 carbon atoms,

R ₇ is a pentafluorophenyl group or succinimide group,

n is an integer of 1-3.

First, the diamine compound (A) is dissolved in a solvent such as acetonitrile, dimethylformamide, etc., and then reacted with alkyl bromoacetate (BrCH ₂ C (= 0) OR ₆ ) in the presence of a base, which is reacted with an amine protecting group. Or protected aminoalkyltosylate to give the corresponding ester compound (B). Diisopropylethylamine, potassium carbonate and the like are used as the base, and benzyloxycarbonyl group, t-butoxycarbonyl group, p-toluenesulfonyl, etc. are used as the amine protecting group, and the protected aminoalkyltosylate. Specific examples of the benzyloxycarbonylaminopropyl tosylate and the like is used. And the temperature range of the reaction is 5 to 80 ℃.

Thereafter, the ester compound (B) is hydrolyzed to obtain a carboxylic acid compound (C). At this time, the hydrolysis reaction is carried out in the presence of a base catalyst such as sodium hydroxide. Subsequently, the corresponding ester compound (D) is obtained through the esterification reaction of the carboxylic acid compound (C) thus formed.

Wherein R ₁ and R ₂ are each independently a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, and R ₃ , R ₄ and R ₅ are each independently a hydrogen atom or carbon number Is selected from the group consisting of 2 to 4 protected aminoalkyl groups, protected diaminoalkyl groups of 5 to 10 carbon atoms, benzyloxycarbonyl groups, t-butoxycarbonyl groups and p-toluenesulfonyl groups, and ○ is a poly (propyleneimine) dendrimer And a dendrimer center selected from the group consisting of poly (amidoamine) dendrimers, Y is O or NH, m is the number of functional groups on the surface according to the number of generations of the dendrimer, an integer from 3 to 200, n is from 1 to 1 Is an integer of 3, x is an integer of 2 to 10, and X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate.

Compound (F) is prepared by reacting one compound selected from compounds (C) and (D) prepared according to Scheme 1 with a functional group present on the surface of the dendrimer (E). In this reaction, the content of compounds (C) and (D) relative to the dendrimer (E) is adjusted taking into account the number of functional groups present on the outer surface in the dendrimer (E). If the dendrimer (E) is reacted with compound (C), various joint reagents can be used. Specific examples of the joint reagent include 1,3-dicyclohexylcarbodiimide (DCC), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (EDCI), and the like. When the dendrimer (E) is reacted with the compound (D), the reaction takes place in the presence of an amine base such as triethylamine or the like.

Thereafter, the deprotection reaction of the protected amine group of the compound (F) is carried out to synthesize a polyammonium dendrimer represented by the formula (1). Although the deprotection reaction of the protected amine group is not particularly limited, it is preferable to carry out using an acid such as hydrochloric acid, bromic acid, trifluoroacetic acid or the like or a mixture thereof in all respects.

The polyammonium dendrimer prepared according to the above-described manufacturing method may form a self-assembled complex through ion interaction with a biochemically active material such as polynucleotide. Therefore, the polyammonium dendrimer of the present invention can be usefully applied to delivery and controlled release of drugs, genes, etc. having the above-described mechanism of action.

For example, when the polynucleotide is used as the biochemically active substance, the polynucleotide is reacted with the polyammonium dendrimer of Formula 1 to form a polynucleotide-polyammonium dendrimer. This dendrimer can be used for the purpose-directed delivery of polynucleotides.

Meanwhile, the pseudorotaxane dendrimer derivative represented by Chemical Formula 2 and a preparation method thereof will be described below.

<Formula 2>

In the above formula, R ₂ is a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms,

Represents a dendrimer center selected from the group consisting of poly (propyleneimine) dendrimers and poly (amidoamine) dendrimers,

Y is an oxygen atom or an amino group,

m is the number of functional groups on the surface according to the generation number of the dendrimer, and is an integer from 3 to 200,

n is an integer of 1 to 3,

x is an integer from 2 to 10,

X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate.

The formation of pseudorotaxane is as follows. It is well known that polyammonium derivatives, especially polyammonium derivatives having 4 to 6 alkyl lengths between two ammonium, readily form pseudorotaxanes by self-assembly with the artificial receptor cucurbityl represented by the formula (3). J. Org. Chem. 1986 , 51 , 4440) Cucurbityl is a compound that has been synthesized 100 years ago and is a macrocyclic compound in which glycoluril is linked to methylene units. The linked macrocyclic compound is shown.

Non-covalent interactions of diammonium alkanes and cucurbityls are due to hydrophobic interactions and hydrogen bonding. The interior of the cucurbitryl is hydrophobic and the carbonyl group at the inlet is the site that provides the hydrogen bond. Thus, as shown in the following figure, the space inside the cucurbityl has a hydrophobic interaction with the alkyl portion of the diammonium alkanes, and the carbonyl group at the inlet of the cucurbitryl is hydrogen-bonded with the ammonium portion of the diammonium alkanes to stabilize the pseudorotaxane. To form a complex.

Thus, the pseudorotaxane dendrimer derivative of Formula 2 is formed by a self-assembly reaction between the polyammonium dendrimer of Formula 1 and the cucurbityl represented by Formula 3.

The self-assembly reaction is carried out by dissolving the polyammonium dendrimer of the formula (1) in water, then adding an excess of cucurbityl to it and stirring the mixture at room temperature. As a result of this self-assembly reaction, a dendrimer coated with pseudorotaxane on the surface can be obtained. At this time, since the pseudo-rotaxane formed on the dendrimer surface forms a hard shell, it is possible to prevent the release of small organic substances, dyes, fluorescent substances, metal ions, bioactive substances, medicines, etc. in the dendrimer.

On the other hand, the pseudo-rotaxane dendrimer represented by the formula (2) can dissociate the cucurbityl by a chemical method. For example, after dissolving the pseudototaxane dendrimer of Formula 2 in water, and then adding a base thereto and stirring for a predetermined time, the cucurbitryl can be dissociated. In this case, as the base, an inorganic base such as sodium bicarbonate, sodium carbonate, potassium bicarbonate, sodium hydroxide, potassium hydroxide, or an organic base such as triethylamine, diisopropylethylamine, etc. is used, and the content thereof is determined according to the number of dendrimer generations. From 10 equivalents to 500 equivalents.

In particular, the pseudorotaxane dendrimer derivative according to the present invention is a compound wherein R ₂ is a hydrogen atom, n is 1 to 3, Y is O or NH, and Br, Cl, PF ₆ or R ₂ is It is preferred that the compound is an aminopropyl group, n is 1 to 3, Y is O or NH, and Br, Cl, PF ₆ .

As described above, using the polyammonium dendrimer or pseudo-rotaxane dendrimer derivative of the formula (1) can obtain a composition for the capture and delivery of biochemically active substances. And various additives conventionally used can be added to this composition.

Hereinafter, the present invention will be described with reference to the following examples, but the present invention is not limited only to the following examples.

Example 1 Preparation of Methyl 3,8- (N, N'-dibenzyloxycarbonyl) -diaminooctylester

To a solution of N-benzyloxycarbonyl diaminobutane dissolved in 60 ml of acetonitrile, diisopropylethylamine (8.7 ml, 50 mmol) and methyl bromoacetate (3.04 g, 20 mmol) were continuously added dropwise at 4 ° C. It was then stirred at room temperature for 12 hours. The reaction mixture was then concentrated and dissolved again with methylene chloride (100 mL), then washed with water and brine and dried.

Thereafter, the dried solution was concentrated to obtain a residue, which was subjected to column chromatography to separate the pure intermediate amine compound. The isolated intermediate amine compound was dissolved in tetrahydrofuran (50 mL) and added to water (50 mL) in which sodium bicarbonate (1.1 equiv.) Was dissolved, and the temperature of the reaction mixture was maintained at 5 ° C. Subsequently, benzyloxycarbonyl chloride (1.1 equiv) was added to the reaction mixture, which was then stirred at 5 ° C. for 2 hours and then at room temperature for 3 hours. Thereafter, the resultant was saturated with salt, ethyl acetate (100 mL) was added, and the organic layer was separated. The organic layer thus separated was washed with 5% hydrochloric acid solution and brine and dried. The dried solution was then concentrated to give 5.14 g (yield: 60%) of methyl 3,8- (N, N'-dibenzyloxycarbonyl) -diaminooctylester.

¹ H NMR (CDCl ₃ ) δ 1.51 (m, 4 H), 3.13 and 3.19 (two t, J = 6.1 and 5.8 Hz, 2 H), 3.32 (t, J = 7.2 Hz, 2 H), 3.62 and 3.70 (two s, 3H), 3.92 and 3.97 (two s, 2 H), 4.79 and 4.97 (two br s, 1 H), 5.06 (s, 2 H), 5.08 and 5.13 (two s, 2 H), 7.32 (m, 5 H);

¹³ C NMR (CDCl ₃ ) δ 25.6, 25.9, 27.4, 41.0, 48.3, 48.9, 49.3, 49.5, 52.5, 52.6, 67.0, 67.8, 67.9, 129.2, 128.36, 128.41, 128.5, 128.8, 128.85, 128.9, 136.9, 137.0, 156.4, 156.8, 170.7.

Example 2. Preparation of 3, 8- (N, N'-dibenzyloxacarbonyl) diaminooctic acid.

A solution of methyl 3,8- (N, N'-dibenzyloxacarbonyl) diaminooctyl ester (2.1 g, 5 mmol) in tetrahydrofuran (30 mL) was dissolved in sodium hydroxide (0.2 g, 5 mmol) in H _2. It was added to an aqueous sodium hydroxide solution prepared by dissolving in O (30 mL). At this time, the temperature of the reaction mixture was maintained at about 5 ℃. The reaction mixture was then stirred at 5 ° C. for 2 hours and then at room temperature for 2 hours.

The reaction mixture was then acidified with 5% hydrochloric acid solution, saturated with salt and extracted with ethyl acetate. The resulting extraction solution was then dried and concentrated to give 1.7 g (yield: 97%) of 3,8- (N, N'-dibenzyloxacarbonyl) diaminooctic acid.

¹ H NMR (CDCl ₃ ) δ 1.51 (m, 4 H), 3.12 and 3.16 (two t, J = 6.1 and 5.6 Hz, 2 H), 3.32 (t, J = 7.1 Hz, 2 H), 3.96 and 4.00 (two s, 2 H), 4.86 (br s, 1 H), 5.06 (s, 2 H), 5.10 and 5.13 (two s, 2 H), 7.31 (m, 5 H), 8.41 (br s, 1 H);

¹³ C NMR (CDCl ₃ ) δ 25.5, 25.8, 27.4, 30.7, 41.0, 48.3, 48.9, 49.0, 49.5, 67.1, 67.7, 67.9, 68.1, 128.1, 128.4, 128.5, 128.8, 128.9, 136.7, 136.9, 156.5, 157.2, 174.2.

Example 3. Preparation of pentafluorophenyl 3, 8- (N, N'-dibenzyloxycarbonyl) dianinooctyl ester.

A solution of 3,8- (N, N'-dibenzyloxycarbonyl) diaminooctic acid (3.5 g, 8.5 mmol) and pentafluorophenol (1.55 g, 8.5 mmol) in ethyl acetate (50 mL) was prepared. After cooling to 5 ° C., 1,3-dicyclohexylcarbodiimide (DCC) (1.74 g, 8.5 mmol) was added thereto. The reaction mixture was then stirred at 5 ° C. for 2 hours and at room temperature for 3 hours.

Thereafter, the suspended solid formed in the reaction mixture was filtered off, and then concentrated to obtain a residue, which was separated by column chromatography to obtain pentafluorophenyl 3,8- (N, N'-dibenzyloxycarbonyl) dia Minotoctyl ester was obtained.

¹ H NMR (CDCl ₃ ) δ 1.56 (m, 4 H), 3.16 and 3.21 (two t, J = 6.2 and 6.3 Hz, 2 H), 3.41 (t, J = 7.7 Hz, 2 H), 4.30 and 4.36 (two s, 2H), 4.76 and 4.95 (br s, 1H), 5.09 (s, 2H), 5.15 and 5.18 (two s, 2H), 7.31 (m, 5H).

The following examples relate to the first-generation dendrimer (G ₁ ) representative reaction, where G _n means the n-generation dendrimer.

Example 4 Preparation of Dendrimer of Structural Formula

Poly (propyleneimine) (PPI) first-generation dendrimer (124 mg, 0.074 mmol) and triethylamine (0.17 mL, 1.24 mmol) were prepared using pentafluorophenyl 3, 8- (N, N'-dibenzyloxycarbonyl) di. Aninooctyl ester (0.7 g, 1.2 mmol) was added to a solution dissolved in methylene chloride (20 mL) and stirred at room temperature for 24 hours.

Thereafter, methylene chloride (50 mL) was added to the reaction mixture, which was washed with water and saturated sodium bicarbonate solution, dried and concentrated. The residue was then separated by column chromatography to give 493 mg (yield: 83%) of the first-generation dendrimer target compound.

G1 : 83% yield ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.52 (m, 28 H), 2.35 (m, 12 H), 3.12 (m, 8 H), 3.22 (m, 8 H), 3.31 ( t, J = 6.9 Hz, 8 H), 3.84 (s, 8 H), 5.06 and 5.09 (two s, 16 H), 5.27 (br s, 4 H), 7.04 (br s, 4 H), 7.31 ( m, 40 H); ESI-MS: m / z: 963.0 [M + H + Na] ²⁺ , 974.0 [M + 2Na] ²⁺ , 982.0 [M + Na + K] ²⁺ , 1903.0 [M + H] ⁺ , 1924.9 [M + Na] ⁺ , 1940.9 [M + K] ⁺ .

G2 : 96% yield ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.42 and 1.50 (m, 60 H), 2.32 (m, 24 H), 2.44 (m, 12 H), 3.10 (m, 16 H), 3.20 (m, 16 H), 3.29 (distorted t, J = 6.6 Hz, 16 H), 3.83 (s, 16 H), 5.04 and 5.07 (two s, 32 H), 5.23 and 5.37 (two br s, 8 H), 7.20 (br s, 8 H), 7.30 (m, 80 H); ESI-MS: m / z: 1984.1 [M + H + Na] ²⁺ , 1995.1 [M + 2Na] ²⁺ .

G3 : 83% yield ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.49 (m, 124 H), 2.31 (m, 52 H), 2.42 (m, 32 H), 3.08 (m, 32 H), 3.20 ( m, 32 H), 3.27 (distorted t, 32 H), 3.83 (s, 32 H), 5.03 and 5.05 (two s, 64 H), 5.29 and 5.42 (two br s, 16 H), 7.24 (br s , 16 H), 7.29 (m, 160 H); ESI-MS: m / z: 2692.0 [M + H + 2Na] ³⁺ , 2699.4 [M + 3Na] ³⁺ .

G4 : 87% yield ¹ H NMR (300 MHz, CDCl ₃ ) δ 1.48 (m, 252 H), 2.32 and 2.48 (m, 180 H), 3.06 (m, 64 H), 3.19 (m, 64 H), 3.25 (m, 64 H), 3.82 (s, 64 H), 5.02 (br s, 128 H), 5.29 and 5.38 (two br s, 32 H), 7.27 (m, 320 H), 7.46 (br, 32 H); ESI-MS: m / z: 4070.6 [M + 4Na] ⁴⁺ , 3263.2 [M + 5Na] ⁵⁺ .

G5 : 87% yield ¹ H NMR (300 MHz, CDCl ₃ ) δ d = 1.41 (m, 508 H), 2.30 and 2.31 (m, 372 H), 3.03 (m, 128 H), 3.22 (m, 256 H ), 3.83 (br s, 128 H), 5.02 (br s, 256 H), 5.51 and 5.63 (two br s, 64 H), 7.29 (m, 640 H), 7.62 (br, 64 H).

Example 5 Preparation of the dendrimer of the following structural formula.

30% bromic acid-acetic acid (10 mL) was added to the first-generation dendrimer (420 mg, 0.05 mmol) obtained in Example 4, which was then stirred at room temperature for 10 hours. Diethyl ether (50 mL) was added to the reaction mixture to form a precipitate. The precipitate was separated by filtration to obtain 370 mg (yield: 95%) of the first-generation dendrimer target compound as a white solid.

G1 : 95% yield ¹ H NMR (300 MHz, D ₂ O) δ 1.82 (m, 2 0 H), 2.01 (m, 8 H), 3.07 (t, J = 6.5 Hz, 8 H), 3.18 (t , J = 7.9 Hz, 8 H), 3.28 (m, 12 H), 3.38 (t, J = 6.8 Hz, 8 H), 3.96 (s, 8 H); ¹³ C NMR (300 MHz, D ₂ O) δ 21.1, 23.1, 23.8, 24.3, 37.0, 39.3, 47.5, 48.5, 51.3, 52.7, 166.6; ESI-MS: m / z: 208.3 [M + 4H] ⁴⁺ , 277.3 [M + 3H] ³⁺ , 415.5 [M + 2H] ²⁺ .

G2 : 96% yield ¹ H NMR (300 MHz, D ₂ O) δ 1.81 (m, 36 H), 2.03 (m, 16 H), 2.31 (m, 8 H), 3.06 (t, J = 6.3 Hz, 16 H), 3.18 (t, J = 7.6 Hz, 16H), 3.34- 3.41 (m, 52H), 3.97 (s, 16H); ¹³ C NMR (300 MHz, D ₂ O) δ 19.5, 23.1, 23.8, 24.3, 36.9, 39.2, 47.4, 48.5, 50.2, 51.4, 166.5; ESI-MS: m / z: 300.9 [M + 6H] ⁶⁺ , 360.6 [M + 5H] ⁵⁺ , 450.8 [M + 4H] ⁴⁺ .

G3 : 95% yield ¹ H NMR (300 MHz, D ₂ O) δ 1.83 (m, 68 H), 2.05 (m, 32H), 2.38 (m, 24 H), 3.08 (t, J = 6.5 Hz, 32 H), 3.21 (t, J = 7.5 Hz, 32H), 3.40-3.49 (m, 116H), 4.00 (s, 32H); ¹³ C NMR (300 MHz, D ₂ O) δ 19.6, 23.1, 23.8,24.3, 37.0, 39.3, 47.5, 48.6, 50.2, 51.5, 166.5; ESI-MS: m / z: 416.4 [M + 9H] ⁹⁺ , 468.3 [M + 8H] ⁸⁺ , 535.0 [M + 7H] ⁷⁺ .

G4 : 96% yield ¹ H NMR (300 MHz, D ₂ O) δ 1.84 (m, 132 H), 2.08 (m, 64 H), 2.39 (m, 56 H), 3.10 (t, J = 6.7 Hz, 64 H), 3.22 (t, J = 7.5 Hz, 64 H), 3.43-3.53 (m, 244 H), 4.02 (s, 64 H); ¹³ C NMR (300 MHz, D ₂ O) δ 19.7, 23.2, 23.8, 24.4, 37.0, 39.3, 47.5, 48.7, 50.3, 51.5, 166.5; ESI-MS: m / z: 847.2 [M + 9H] ⁹⁺ , 762.6 [M + 8H] ⁸⁺ , 693.5 [M + 7H] ⁷⁺ , 635.6 [M + 6H] ⁶⁺ .

G5: 97% yield^OneH NMR (300 MHz, D₂O): δ 1.84 (m, 260 H), 2.07 (m, 128 H), 2.38 (m, 120 H), 3.09 (t, J = 6.5 Hz, 128 H), 3.22 (distorted t, 128 H), 3.42-3.53 (m, 500 H), 4.02 (s, 128 H);¹³C NMR (300 MHz, D₂O) δ 20.8, 23.2, 23.8, 24.4, 37.0, 39.3, 47.5, 48.7, 50.3, 51.4, 166.4.

Example 6. Preparation of pseudorotaxane dendrimer of the following structural formula.

Cucurbitryl (128 mg, 0.13 mmol) was added dropwise thereto while stirring the solution of the first-generation polyammonium dendrimer (50 mg, 6.7 μm) obtained in Example 5 in water (30 mL). Subsequently, the reaction mixture was stirred at room temperature for 10 hours, and excess cookerbitril was filtered off.

Thereafter, the reaction mixture in a clear solution state was concentrated to a solution of about 2 mL, and then ethanol (30 mL) was added to form a precipitate. The precipitate thus obtained was separated by filtration to obtain 152 mg (yield: 97%) of the first compound of the similar rotataxane dendrimer as a white solid.

G1 : 97% yield ¹ H NMR (300 MHz, D ₂ O) δ 0.58-0.68 (m, 16 H), 1.92 (m, 4 H), 2.11 (m, 8 H), 2.31-2.36 (m, 16 H), 3.36 (m, 12 H), 3.47 (m, 8 H), 4.17 (s, 8 H), 4.40-4.49 (m, 48 H), 5.72-5.80 (m, 96 H); ESI-MS: m / z: 536.2 [M + 9H] ⁹⁺ , 603.1 [M + 8H] ⁸⁺ , 689.0 [M + 7H] ⁷⁺ , 700.7 [M + 7H + HBr] ⁷⁺ , 712.1 [M + 7H + 2HBr] ⁷⁺ , 830.7 [M + 6H + 2HBr] ⁶⁺ , 844.2 [M + 6H + 3HBr] ⁶⁺ , 858.0 [M + 6H + 4HBr] ⁶⁺ , 1029.0 [M + 5H + 4HBr] ⁵⁺ , 1045.5 [M + 5H + 5HBr] ⁵⁺ .

G2 : 97% yield ¹ H NMR (300 MHz, D ₂ O) δ 0.57-0.67 (m, 32 H), 1.92 (m, 4 H), 2.14 (m, 24 H), 2.28-2.34 (m, 32 H), 3.20-3.47 (m, 52 H), 4.17 (s, 16 H), 4.43-4.53 (m, 96 H), 5.75-5.81 (m, 192 H).

G3 : 97% yield ¹ H NMR (300 MHz, D ₂ O) δ 0.60-0.67 (m, 64 H), 1.92-2.14 (m, 36 H), 2.31 (m, 88 H), 3.20-3.44 (m 116 H), 4.15 (s, 32 H), 4.44-4.55 (m, 192 H), 5.73-5.76 (m, 384 H).

G4 : 98% yield ¹ H NMR (300 MHz, D ₂ O) δ 0.63-0.67 (m, 128 H), 1.92-2.12 (m, 132 H), 2.31 (m, 184 H), 2.97-3.43 (m , 244 H), 4.16 (s, 64 H), 4.47-4.52 (m, 384 H), 5.72-5.78 (m, 768 H).

G5 : 97% yield ¹ H NMR (500 MHz, D ₂ O, 50 ^o C): δ 0.97 and 1.06 (m, 256 H), 2.58 (m, 300 H), 2.67 (m, 376 H), 3.85 ( m, 500 H), 4.51 (s, 128 H), 4.89-4.96 (m, 496 H), 6.11-6.20 (m, 1536 H).

Application example. A study on the dissociation of pseudorotaxane dendrimer.

The pseudorotaxane dendrimer (5 mg) obtained according to Example 6 was dissolved in heavy water (D ₂ O) (0.6 ml) and observed by nuclear magnetic resonance analysis, and sodium hydroxide (5 mg) was added to the aqueous solution of the pseudorotaxane dendrimer. After addition and stirring, it was observed again by nuclear magnetic resonance analysis.

As a result of analysis by nuclear magnetic resonance analysis, it was confirmed that the addition of a base to the aqueous solution of pseudorotaxane dissociates the cucurbityl forming the pseudorotaxane. This is an example showing that the trapped material can be released by dissociating the pseudorotaxane when it is trapped inside the dendrimer.

The polyammonium dendrimer of the formula (1) according to the present invention has a polyamine group at the terminal to react with a receptor such as cucurbityl to form a self-assembled complex such as a pseudorotaxane dendrimer derivative of the formula (2) to form a biochemically active material inside the dendrimer Capable of trapping and dissociating the formed complexes can release the biochemically active substances collected therein slowly, and the polyammonium dendrimer of formula 1 is self-assembled through ionic interactions with biochemically active substances such as polynucleotides. Complexes may be formed. Therefore, such materials can be usefully used in biochemical fields such as drug delivery, gene therapy, gene delivery, and the like.

Although the present invention has been described with reference to the embodiments, these are merely exemplary, and those skilled in the art will understand that various modifications and equivalent implementations are possible therefrom. Therefore, the true technical protection scope of the present invention will be defined by the technical spirit of the appended claims.

Claims (16)

Polyammonium dendrimer represented by Formula 1: <Formula 1> In the above formula, R ₁ and R ₂ are each independently a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, Represents a dendrimer central region selected from the group consisting of poly (propyleneimine) dendrimers and poly (amidoamine) dendrimers, Y is O or NH, m is the number of functional groups on the surface according to the generation number of the dendrimer, and is an integer from 3 to 200, n is an integer of 1 to 3, x is an integer from 2 to 10, X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate. The polyammonium dendrimer according to claim 1, wherein R ₁ and R ₂ are hydrogen atoms, n is 1 to 3, Y is O or NH, and X is Br, Cl, PF ₆ . The compound according to claim 1, wherein R ₁ is a hydrogen atom, R ₂ is an aminopropyl group, n is 1 to 3, Y is O or NH, and X is Br, Cl, PF ₆ Polyammonium dendrimer. The polyammonium dendrimer according to claim 1, wherein R ₁ and R ₂ are aminopropyl groups, n is 1 to 3, Y is O or NH, and X is Br, Cl, PF ₆ . The compound according to claim 1, wherein R ₁ is an aminopropyl group, R ₂ is a hydrogen atom, n is 1 to 3, Y is O or NH, and X is Br, Cl, PF ₆ Polyammonium dendrimer. (a) reacting a diamine compound (A) with an alkyl bromoacetate and reacting it with an amine protecting group or with a protected aminoalkyltosylate to obtain an ester compound (B); (b) hydrolyzing the ester compound (B) to obtain a carboxylic acid compound (C); (c) obtaining an ester compound (D) by esterification of the carboxylic acid compound (C); (d) reacting a dendrimer (E) with one selected from the carboxylic acid compound (C) and the ester compound (D) to obtain a compound (F); And (e) conducting a deprotection reaction of the protected amine group of compound (F); a method of producing a polyammonium dendrimer represented by the formula (1), including the above. In the above formula, R ₁ and R ₂ are each independently a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, R ₃ , R ₄ and R ₅ independently of one another are a hydrogen atom, a protected aminoalkyl group having 2 to 4 carbon atoms, a protected diaminoalkyl group having 5 to 10 carbon atoms, benzyloxycarbonyl group, t-butoxycarbonyl group and p-toluenesul Selected from the group consisting of a polyyl group, R ₆ is an alkyl group having 1 to 5 carbon atoms, R ₇ is a pentafluorophenyl group or succinimide group, Represents a dendrimer center selected from the group consisting of poly (propyleneimine) dendrimers and poly (amidoamine) dendrimers, Y is O or NH, m is the number of functional groups on the surface according to the generation number of the dendrimer and is an integer from 3 to 200, n is an integer of 1 to 3, x is an integer from 2 to 10, X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate. The method of claim 6, wherein in step (d), when the dendrimer (E) is reacted with the carboxylic acid compound (C), 1,3-dicyclohexylcarbodiimide (DCC) and 1- (3-dimethyl A process for producing a polyammonium dendrimer, characterized in that a joint reagent selected from the group consisting of aminopropyl) -3-ethylcarbodiimide (EDCI) is used. The method of claim 6, wherein in the step (d), when the dendrimer (E) is reacted with the ester compound (D), the polyammonium dendrimer is produced in the presence of an amine base. The method of claim 6, wherein the deprotection of (e) is carried out using at least one acid selected from the group consisting of hydrochloric acid, bromic acid and trifluoroacetic acid. Pseudorotaxane dendrimer derivatives represented by Formula 2; <Formula 2> In the above formula, R ₂ is a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, Represents a dendrimer center selected from the group consisting of poly (propyleneimine) dendrimers and poly (amidoamine) dendrimers, Y is an oxygen atom or an amino group, m is the number of functional groups on the surface according to the generation number of the dendrimer, and is an integer from 3 to 200, n is an integer of 1 to 3, x is an integer from 2 to 10, X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate. 11. The pseudorotaxane dendrimer derivative according to claim 10, wherein R ₂ is a hydrogen atom, n is 1 to 3, Y is O or NH, and X is Br, Cl, PF ₆ . The pseudorotaxane dendrimer derivative according to claim 10, wherein R ₂ is an aminopropyl group, n is 1 to 3, Y is O or NH, and X is Br, Cl, PF ₆ . A method for producing a pseudo-rotaxane dendrimer derivative represented by the formula (2) characterized by adding an excess of cucurbityl to the polyammonium dendrimer of the formula (1). <Formula 1> <Formula 2> In the above formula, R ₁ and R ₂ are each independently a hydrogen atom, an aminoalkyl group having 2 to 4 carbon atoms or a diaminoalkyl group having 5 to 10 carbon atoms, Represents a dendrimer central region selected from the group consisting of poly (propyleneimine) dendrimers and poly (amidoamine) dendrimers, Y is O or NH, m is the number of functional groups on the surface according to the generation number of the dendrimer, and is an integer from 3 to 200, n is an integer of 1 to 3, x is an integer from 2 to 10, X is selected from the group consisting of halogen atom, hexafluorophosphate, trifluoroacetate. The method of claim 13, wherein the reaction is performed by dissolving the polyammonium dendrimer in water and then adding an excess of cucurbityl and stirring at room temperature. A composition for capturing and delivering a biochemically active substance, comprising the polyammonium dendrimer according to any one of claims 1 to 5. A composition for capturing and delivering a biochemically active substance, comprising the pseudorotaxane dendrimer derivative according to any one of claims 10 to 12.