WO2010105450A1

WO2010105450A1 - A layered double hydroxide complex inserted by carnosine and the preparation method thereof

Info

Publication number: WO2010105450A1
Application number: PCT/CN2009/071925
Authority: WO
Inventors: 卫敏; 陈秋华; 陆军; 段雪
Original assignee: 北京化工大学
Priority date: 2009-03-18
Filing date: 2009-05-22
Publication date: 2010-09-23
Also published as: CN101530621A

Abstract

A layered double hydroxide complex inserted by carnosine and the preparation method thereof are provided. The preparation method including codeposition method and ion exchange method, which use microwave hydrothermal step for shorter reaction time, is disclosed.

Description

Carnosine drug intercalated hydrotalcite and preparation method thereof

Technical field:

The invention belongs to the technical field of dipeptide drugs. In particular, the present invention provides mesopeptide intercalated hydrotalcite and a process for its preparation. Since the microwave hydrothermal method is employed in the preparation process of the present invention, the crystallization time of the coprecipitation method and the reaction time of the ion exchange method are remarkably shortened. Background technique:

Carnosine is a natural water-soluble dipeptide substance present in the skeletal muscle of vertebrates. Its structural composition is β-alanine-L-histidine, and its molecular weight is 226.24. The physiological properties of carnosine, such as buffering, oxidation resistance, activity of neurotransmitters, and regulators of some enzymes, have been extensively studied. Carnosine is currently known to have a good application in the food additive and cosmetic industries.

However, due to their sensitivity to chemicals and enzymes, the use of dipeptides as a drug in clinical treatment is limited. In recent years, carnosine has made breakthroughs in the treatment of chronic diseases caused by low antioxidant function, such as cataracts and gastric ulcers. Harbin Medical University has reported that carnosine has a therapeutic effect on various types of cataracts (effectiveness of 80%). Several possible routes of dipeptide administration have been reported, such as oral, pulmonary inhalation, mucosal or subcutaneous administration, which typically require specific transport devices and/or penetration promotion and help dipeptide drugs from the point of release. Enter the body's circulatory system. The main goal of these improvements is not only to overcome the lack of bioavailability after oral administration, but also to avoid subcutaneous injections with poor patient compliance.

Bimetallic composite hydroxides, also known as Layered Double Hydroxides (LDHs), are a new type of multifunctional layered material with good chemical stability, strong heat resistance, and laminate metal of LDHs. The ion species and ratio can be adjusted and changed within a certain range, and the interlayer anions are exchangeable. The use of hydrotalcite in this property can be carried with hydrophilic side groups The dipeptide (such as coo_) is inserted into the layer of hydrotalcite by ion exchange to form a supramolecular structure system, which is effective for improving the stability of the dipeptide. The function of hydrotalcite as a "molecular container" will make it a novel carrier for the storage and release of dipeptide drugs, and at the same time contribute to the development of new and effective routes of administration for dipeptide drugs. At present, there are no literatures and patent reports on the combination of hydrotalcite-like layered materials and carnosine. Summary of the invention:

The object of the present invention is to provide a dipeptide drug intercalated hydrotalcite system and a preparation method thereof. The present invention not only improves the stability of carnosine, but also applies hydrotalcite as a novel carrier to the storage and release of dipeptide drugs, and contributes to the development of a new and effective route of administration of dipeptide drugs.

In the dipeptide drug intercalation hydrotalcite system of the invention, the hydrotalcite acts as a drug carrier and controls the release of the drug, and therefore, the space confinement between the hydrotalcite layers can be maintained during storage to maintain the two inserted therein. The structure of the peptide drug is stable, and the effect of sustained release of the drug is exerted by the interaction between the host and the guest of the hydrotalcite layered composite. Specifically, the dipeptide drug intercalated hydrotalcite according to the present invention is an anionic supramolecular layered material obtained by a coprecipitation method or an ion exchange method, and in the anionic supramolecular layered material, a dipeptide drug anion accounts for 20%-80% of the total number of moles of anions in the hydrotalcite layer.

The carnosine intercalated hydrotalcite of the present invention can be produced by a coprecipitation method or an ion exchange method.

The preparation steps of the coprecipitation method are as follows:

A. preparing a nitrate mixed solution of soluble magnesium nitrate and soluble aluminum nitrate, wherein the divalent metal magnesium has an ion concentration of 0.8-1.6 M, and the magnesium and aluminum metal ion molar ratio ranges from 2-3;

B. preparing a mixed solution of the interlayer guest (L-carnosine) and NaOH;

C. The nitrate mixed solution prepared in the step A is slowly added dropwise to the mixed solution prepared in the step 在 under the condition of the protection, and stirred, and the pH range of the obtained solution is adjusted to 1-5 mol/L NaOH to 7-10, deionized with CO ₂ removed after crystallization at 60 ° C - 70 ° C for 24 hours The water was washed by centrifugation to neutrality, and dried at 70 ° C for 12-24 hours to obtain a carnosine intercalated hydrotalcite. Wherein, the temperature of the deionized water from which CO ₂ is removed may be 40-80 ° C. Preferably, the temperature of the deionized water from which CO ₂ is removed may be 60-70 ° C.

The amount of the nitrate mixed solution prepared in the step A used in the step C and the mixture prepared in the step B is such that the anion of the carnosine accounts for 20-80% of the total number of interlayer anions in the obtained anionic supramolecular layered material. .

The preparation steps of the ion exchange method are as follows:

A. preparing a nitrate mixed solution of soluble magnesium nitrate and soluble aluminum nitrate, wherein the ion concentration of the divalent metal magnesium is 0.8-1.6 M, and the molar ratio of magnesium and aluminum metal ions is 2-3;

B. Preparing an NaOH alkaline solution;

C. The nitrate mixed solution prepared in step A is slowly added dropwise to the alkali solution prepared in step B under N ₂ protection conditions, stirred, and the pH range of the obtained solution is obtained by using 1-5 mol/L NaOH. Adjusted to 8-10, crystallized at 60 ° C -100 ° C for 12-48 hours, then centrifuged with deionized water with CO ₂ removed to neutrality to obtain a nitrate hydrotalcite precursor; wherein, the removal The temperature of the deionized water of C0 ₂ may be 40-80 ° C. Preferably, the temperature of the deionized water from which C0 ₂ is removed is 60-70 ° C;

D. The carnosine is dissolved in deionized water, and the undried nitrate hydrotalcite precursor is obtained by adding step C, and ion exchange is carried out in a water bath at 60 ° C under N ₂ protection for 12-24 hours, and C0 is removed. ₂ , deionized water was centrifuged and washed to neutrality, and dried at 70 ° C for 12-24 hours to obtain intercalated hydrotalcite with interlayer material as carnosine. The amount of carnosine and nitrate hydrotalcite precursor was such that the anion produced was obtained. In the supramolecular layered material, the anion of carnosine accounts for 20-80% of the total number of moles of interlayer anions. Specifically, the amount of carnosine dissolved per 100 mL of deionized water is 0.05-4 g, preferably 1-2 g (0.0045-0.009 mol), and step C is added to obtain an undried nitrate hydrotalcite precursor. The amount is 10-20 g, preferably 5-10 g. Preferably, the mass ratio of carnosine to nitrate hydrotalcite precursor is (0.5-1.5): 3. Wherein, the temperature of the deionized water from which C0 ₂ is removed may be 40-80 ° C, Preferably, the temperature of the deionized water from which CO ₂ is removed is 60-70 ° C.

X-ray diffraction (XRD), infrared spectroscopy (IR), nuclear magnetic resonance (NMR), and elemental analysis of the materials prepared above showed that the carnosine was successfully inserted into the hydrotalcite layer; in situ XRD (In-situ XO), original The IR (In-situ l), thermogravimetry-differential heat (TG-DTA), thermogravimetry-mass analysis (TG-MAS) characterization showed that the thermal stability of the composite was significantly improved; The composite system can achieve a certain sustained release performance.

Further, the present invention preferably employs a microwave hydrothermal method in the preparation of the myopeptide intercalated hydrotalcite, so that the method according to the present invention can significantly shorten the crystallization time of the coprecipitation method and the reaction time for performing the ion exchange method; The prepared dipeptide drug (gp, carnosine) intercalated hydrotalcite composite material has better thermal stability under normal conditions due to utilizing the spatial confinement effect of hydrotalcite and the interaction between host and guest. Certain sustained release properties facilitate the storage of dipeptide drugs and open up new routes of administration. BRIEF DESCRIPTION OF THE DRAWINGS:

1 is an XRD spectrum obtained under the condition of the specific embodiment 5 of the present invention; the abscissa is 2 Θ, the unit is: degree; the ordinate is intensity;

Figure 2 is a FT-IR spectrum obtained under the conditions of Specific Example 5 of the present invention; the abscissa is the wave number, the unit is cm; and the ordinate is the absorption intensity. detailed description:

The invention is specifically illustrated by the following examples.

In the following examples, the products obtained in the respective examples were determined using the following method:

The structure was analyzed by a Shimadzu XRD-6000 X-ray diffractometer, Cu Ka source (λ = 0.154 nm), voltage 40 Kv, current 30 mA, continuous scanning, scanning speed 5 min.

FT-IR spectra were obtained on VECTOR 22 (Brook, Germany), and samples were mixed with KBr After pressing, the film was scanned at room temperature under an air atmosphere. The parameter indicators are: resolution of 4 cm - ¹ and scanning range of 4000-400 cm. Example 1

Step A: 3.846 g (0.015 mol) of solid Mg(N0 ₃ ) ₂ *6H ₂ 0 and 2.8135 g (0.0075 mol) of solid Α1(Ν0 ₃ ) ₃ ·9Η ₂ 0 were dissolved in 50 mL to remove C0 ₂ Deionized water (solution I); another 4 g (about 0.018 mol) of carnosine and 2.5 g (0.0625 mol) of NaOH dissolved in 50 mL of deionized water removed in CO ₂ (solution II

Step B: The solution II was placed in a four-necked flask, and under the condition of N ₂ gas protection, the solution I was slowly added dropwise to the solution II while vigorously stirring, and the solution was dropped in about 0.5 h. After the completion of the dropwise addition, the pH was adjusted to 7-10 with a 5 mol/L NaOH solution;

Step C: The solution obtained in the step B is subjected to a crystallization reaction in a water bath at 60 ° C for 24 hours; Step D: centrifugation at 60 ° C with deionized water from which 0 ₂ is removed until the pH is about 7 and then at 70 ° C. After drying for 12-24 hours, a magnesium-aluminum myopeptide intercalated hydrotalcite composite material was obtained.

From the XRD spectrum, FT-IR spectrum, NMR and elemental analysis, it was found that the obtained anion of the hydrotalcite layer was an angiopeptide anion. Example 2

In the same manner as in Embodiment 1, the difference is:

The solution obtained in the step B in the first embodiment was transferred to a microwave rapid digestion tank (WX-4000 produced by Shanghai Shuguang Analytical Instruments Co., Ltd.), and the temperature was controlled by microwave heating at 80-100 ° C for 0.5 h. the product was 70 ° C deionized water to remove the centrifuge ₀₂ was washed to pH 7, dried at 70 ° C for 12-24h.

From the XRD spectrum, FT-IR spectrum, NMR and elemental analysis, it was found that the anion of the hydrotalcite layer was an angiopeptide anion. Example 3

Step A: Weigh 15.384 g of Mg(N0 ₃ ) ₂ ^63⁄40 and 11.2539 g of A1(N0 ₃ ) ₃ <9H ₂ 0 dissolved in 100 mL of deionized water with 0 ₂ removed to prepare a mixture (salt solution i) Another 7.2 g of NaOH was dissolved in 100 mL of deionized water with CO ₂ removed and placed in a 500 mL four-necked flask (alkaline solution ii);

Step B: Slowly drip the salt solution i into the alkali solution ii in a 70 ° C water bath under N ₂ protection and vigorous stirring, and adjust the pH to about 10.0 with 2 mol/L sodium hydroxide. Crystallization reaction for 40 h;

Step C: The product obtained in Step B was washed with 40 of deionized water with CO ₂ removed to pH <8.0; a large amount of fresh filter cake was reserved for ion exchange, and only a small amount of sample was taken and dried at 70 ° C for 18 h. After XRD, it was confirmed that N0 ₃ -Mg ₂ Al-LDHs was obtained, of which Mg ²⁺ /Al ³⁺ =2 _;

Step D: 4 g (0.018 mol) of carnosine was dissolved in 200 mL of deionized water with CO ₂ removed and placed in a 500 mL four-necked flask, and 12 g of the filter cake obtained in the step C was added thereto. The pH was adjusted to 7-9 with 2 mol/L sodium hydroxide;

Step E: Ion exchange was carried out for 12-24 h under the protection and vigorous stirring and the temperature was controlled at 60 ° C. The obtained product was washed with 60 ° C of deionized water with CO ₂ removed and centrifuged. 3-6 times, the obtained precipitate was dried in a vacuum drying oven to obtain a layered composite material in which intermusing anion was interposed.

From the XRD spectrum, FT-IR spectrum, NMR and elemental analysis, it was found that the obtained anion of the hydrotalcite layer was an angiopeptide anion. Example 4

According to the method of Embodiment 3, the difference is:

Transfer the mixed solution obtained in step D of Example 3 to a microwave rapid digestion tank (Shanghai In WX-4000 produced by Dawning Analytical Instruments Co., Ltd., the temperature was controlled by microwave heating at 80-100 °C for 0.5 h, and the obtained product was centrifuged to pH at 70 ° C with deionized water removed with CO ₂ . It is about 7, dried at 70 ° C for 12-24 h.

From the XRD spectrum, FT-IR spectrum, NMR and elemental analysis, it was found that the obtained anion of the hydrotalcite layer was an angiopeptide anion. Example 5

Step A: The mixed salt solution i in the step A of Example 3 was added all at once to the alkali solution ii under high-speed stirring, and after stirring for 20 minutes, the obtained product was washed with deionized water from which C0 ₂ was removed. And centrifuged 3-6 times;

Step B: taking 12 g of the precipitate obtained in step A and 4 g of carnosine dissolved in deionized water with CO ₂ removed, and adjusting the pH to 7-9 with 2 mol/L sodium hydroxide;

Step C: The solution obtained in step B is transferred to an autoclave, and reacted at a constant temperature of 90 ° C for 16 h, and the pressure in the autoclave is 1-10 Mpa;

Step D: The product was washed with deionized water from which CO ₂ was removed and centrifuged for 3 to 6 times, and the precipitate was dried in a vacuum at room temperature to obtain a layered composite material having intermused anion peptide intercalated therebetween.

From the XRD spectrum, FT-IR spectrum, NMR and elemental analysis, it was found that the obtained anion of the hydrotalcite layer was an angiopeptide anion.

Claims

Claim

A mesopeptide intercalated hydrotalcite composite material, characterized in that: the carnopeptide intercalated hydrotalcite composite material is an anionic supramolecular layered material obtained by a coprecipitation method or by an ion exchange method, the anionic supramolecular layer In the material, the anion of carnosine accounts for the total number of moles of interlayer anions.

20-80%; The composite material can make full use of the spatial confinement between the layers and the interaction between the host and the guest, and generally has better thermal stability and a certain sustained release property.

2. The composite material according to claim 1, wherein the coprecipitation method comprises: a. preparing a nitrate mixed solution of soluble magnesium nitrate and soluble aluminum nitrate, wherein the ion concentration of the divalent metal magnesium is 0.8- 1.6 M, magnesium, aluminum metal ion molar ratio range of 2-3;

b. preparing a mixture of interlayer L-carnosine and NaOH;

c The nitrate mixed solution prepared in step a is slowly added dropwise to the mixed solution prepared in step b under N ₂ protection conditions, stirred, and the pH range of the obtained solution is obtained by using 1-5 mol/L NaOH. Adjust to 7-10, crystallize at 60 ° C -70 ° C for 24 hours, then centrifuge to remove neutral with deionized water with CO ₂ removed to neutral, and dry at 70 ° C for 12-24 hours to obtain carnosine intercalation. Hydrotalcite.

The composite material according to claim 2, wherein a microwave hydrothermal method is used in the crystallization of the step c in the coprecipitation method.

4. The composite material according to claim 1, wherein the ion exchange method comprises: a. preparing a nitrate mixed solution of soluble magnesium nitrate and soluble aluminum nitrate, wherein the ion concentration of the divalent metal magnesium is 0.8- 1.6 M, magnesium, aluminum metal ion molar ratio range of 2-3;

b. preparing an alkali solution of NaOH;

c slowly adding the nitrate mixed solution prepared in step a to the alkali solution prepared in step b under the conditions of the protection, stirring, and using the 1-5 mol/L NaOH to adjust the pH of the obtained solution. The range is adjusted to 8-10, and after crystallization at 60 ° C - 100 ° C for 12-48 hours, it is centrifugally washed with deionized water with CO ₂ removed to neutrality to obtain a nitrate hydrotalcite precursor;

d. The carnosine is dissolved in water, and the undried nitrate hydrotalcite precursor obtained in the step c is added. The amount of the carnosine and the nitrate hydrotalcite precursor is such that the obtained anionic supramolecular layer material is carnosine. The anion accounts for 20-80% of the total number of interlayer anions, is ion exchanged in a water bath at 60 ° C and under N ₂ protection for 12-24 hours, and then washed by centrifugation with deionized water with CO ₂ removed to neutral. After drying at 70 ° C for 12-24 hours, an intercalated hydrotalcite having an interlayer material of carnosine was obtained.

The composite material according to claim 4, wherein a microwave hydrothermal method is used in the ion exchange process of the step d in the ion exchange method.

6. Composite material according to claim 4 or 5, characterized in that during the ion exchange of step d of the ion exchange process, the pressure is between l and 10 Mpa.

A method for producing the dipeptide drug intercalated hydrotalcite according to claim 1, wherein the method comprises:

Step a: preparing a mixed salt solution of soluble magnesium nitrate and soluble aluminum nitrate, wherein the divalent metal magnesium has an ion concentration of 0.8-1.6 M, and the magnesium and aluminum metal ion molar ratio ranges from 2-3, and the NaOH alkali solution is prepared and mixed. The salt solution is rapidly added to the alkali solution under high-speed stirring, and after stirring for 20 minutes, the obtained product is washed with deionized water from which CO ₂ is removed and centrifuged for 3-6 times;

Step b: taking an appropriate amount of the precipitate obtained in step a and an appropriate amount of carnosine dissolved in deionized water from which 0 ₂ is removed, adjusting the pH to 7-9, the amount of the precipitate and carnosine is such that the obtained anionic supramolecular In the layered material, the anion of carnosine accounts for 20-80% of the total number of moles of interlayer anions;

Step c: Transfer the solution obtained in step b to an autoclave and incubate at 90 ° C for 16 h; Step d: Wash the product with deionized water with CO ₂ removed and centrifuge for 3-6 times to precipitate The layered composite material in which the carnosine anion is interposed between the layers can be obtained by drying at room temperature in a vacuum.

The method for intercalating hydrotalcite with a dipeptide drug according to claim 7, wherein the pH is adjusted to 7-9 with 1-5 mol/L of sodium hydroxide in the step b.

The method for intercalating hydrotalcite with a dipeptide drug according to claim 7 or 8, wherein the pressure in the autoclave in the step c is 1-10 Mpa.