WO2008086716A1

WO2008086716A1 - Layered double hydroxide complex inserted by dipeptiven and the preparation method thereof

Info

Publication number: WO2008086716A1
Application number: PCT/CN2007/070130
Authority: WO
Inventors: Min Wei; Fengjie Zhu; Xue Duan
Original assignee: Beijing University Of Chemical Technology
Priority date: 2007-01-16
Filing date: 2007-06-18
Publication date: 2008-07-24
Also published as: CN100998872A; CN100998872B

Abstract

A layered double hydroxide complex inserted by dipeptiven and the preparation method thereof are provided. The chemical formula of the layered double hydroxide complex inserted by dipeptiven is: [M2+1-xM3+x(OH)2](A-)x•mH2O; where M2+ is one or more divalent metallic ions selected from Mg2+, Co2+, Ni2+, Ca2+, Cu2+, Fe2+, Mn2+; M3+ is one or more trivalent metallic ions selected from Al3+, Cr3+, Ga3+, In3+, Co3+, Fe3+, V3+; A- is dipeptiven anion inserted into layered double hydroxides; x is a rational number between 0.2-0.33; m is a rational number between 0-2. The layered double hydroxide complex inserted by dipeptiven can be prepared by means of co-precipitation or ion exchange process.

Description

Force peptide intercalated hydrotalcite composite and preparation method thereof

Technical field

The invention relates to a polypeptide intercalation hydrotalcite composite and a preparation method thereof, in particular to a force peptide intercalated hydrotalcite composite and a preparation method thereof. Background technique

Peptides are one of the important life material foundations, and their role involves all aspects of the life process. Peptides affect many important physiological and biochemical functions in living organisms. In many cases, peptides play an irreplaceable role. More than 100 active peptides are known to play a role in the central and peripheral nervous systems, the cardiovascular system, the immune system, and the intestine. Peptides also interact with receptors to affect the exchange of information between cells and participate in many biochemical processes such as metabolism, pain, regeneration, and immune responses.

However, due to chemosensitivity and enzyme sensitivity, the use of peptides as a drug in clinical treatment is limited. In recent years, advances in the field of formulation and delivery systems have led to several highly successful peptide drugs. 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. Several possible routes of polypeptide delivery have been reported, such as oral sputum, pulmonary inhalation, mucosal or subcutaneous administration, which typically require specific transport devices and/or penetration enhancers to aid in drug release. Point into the system loop.

Bimetallic composite hydroxide, also known as Layered Double Hydroxides (LDHs), is a new type of multifunctional layered material with good chemical stability, strong heat resistance, and LDHs laminate metal. The type and ratio of ions can be changed, and the interlayer anions are exchangeable. By using this property, a polypeptide having a hydrophilic side group COO- can be inserted into the interlayer by ion exchange to form a supramolecular structure system, which can effectively improve the stability of the polypeptide. The function of hydrotalcite as a "molecular container" will make it a novel carrier for the storage and release of peptide drugs, and at the same time contribute to the development of new and effective routes of administration for polypeptide drugs. Lipopeptide is a small peptide condensed by alanine and glutamine. It is used in the infusion of parenteral nutrition. It is a supplement to amino acid solution. It is clinically used in patients who need to supplement glutamine, including those in catabolism and high. A patient with a metabolic condition. However, there are no literatures and patent reports on the combination of hydrotalcite-like layered materials and force peptides at home and abroad. Summary of the invention

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

The force peptide intercalated hydrotalcite complex of the present invention maintains the structural stability of the peptide drug through the spatial confinement between the hydrotalcite layers during its storage process, and acts as a drug through the interaction between the host and the guest of the hydrotalcite layered composite material. The effect of slow release, while hydrotalcite also acts as a drug carrier and controls drug release.

The present invention provides a force peptide intercalated hydrotalcite complex, the chemical formula of the hydropeptide intercalated hydrotalcite complex is:

Wherein M ²⁺ represents a divalent metal ion, and may be, for example, any one or more of Mg ²⁺ , Co ²⁺ , Ni ²⁺ , Ca ²⁺ , Cu ²⁺ , Fe ^{2+ ,} and Mn ²⁺ . Preferably, Ni ²⁺ , Co ²⁺ or Mg ²⁺ ; M ³⁺ represents a trivalent metal ion, and may be, for example, Al ³⁺ , Cr ³⁺ , Ga ³⁺ , In ³⁺ , Co ³⁺ , Fe ^{3+ ,} and Any one or more of V ³⁺ , preferably Al ³⁺ ; A—is a force peptide anion as an anion between hydrotalcite layers; X ranges from 0.2 to 0.33, and m ranges from 0 to 2.

The force peptide intercalated hydrotalcite complex of the present invention can be prepared by a coprecipitation method or an ion exchange method. The coprecipitation method includes the following steps:

A. preparing a mixed solution of a soluble divalent metal salt and a soluble trivalent metal salt, wherein the divalent metal ion concentration is 0.01-1.0 M, and the molar ratio of the divalent metal ion to the trivalent metal ion is 2-4; The metal ion may be any of Mg ²⁺ , Co ²⁺ , Ni ²⁺ , Ca ²⁺ , Cu ²⁺ , Fe ^{2+ ,} and Mn ²⁺ . One or more, preferably M ²⁺ , Co ²⁺ or Mg ²⁺ ; the trivalent metal ions may be Al ³⁺ , Cr ³⁺ , Ga ³⁺ , In ³⁺ , Co ³⁺ , Fe ³⁺ and Any one or more of V ³⁺ , preferably Al ³⁺ ;

B. preparing a mixed solution of L-alanyl-L-glutamine and NaOH;

C. The 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 of the mixed solution is adjusted to 7-10 by using NaOH solution. Crystallization at 60 ° C - 70 ° C for 2-24 hours, washing with water to neutral, drying at 50-70 ° C for 12-24 hours, to obtain a force peptide intercalated hydrotalcite complex.

In the step C, the concentration of the NaOH solution for adjusting the pH is preferably 1-5 mol/L; the water used for the centrifugal washing is preferably deionized water with C0 ₂ removed (the water temperature may be 30-65 ° C) ) to avoid the effect of carbonate ions on the resulting product.

In the crystallization process of the above step C, it is preferred to use a microwave hydrothermal method to shorten the crystallization time.

The ion exchange method includes the following steps:

A. preparing a mixed solution of a soluble divalent metal salt and a soluble trivalent metal salt, wherein the divalent metal ion concentration is 0.01-1.0 M, and the molar ratio of the divalent metal ion to the trivalent metal ion is 2-4; The metal ion may be any one or more of Mg ²⁺ , Co ²⁺ , Ni ²⁺ , Ca ²⁺ , Cu ²⁺ , Fe ^{2+ ,} and Mn ²⁺ , preferably M ²⁺ , Co ²⁺ or Mg ²⁺ ; the trivalent metal ion may be any one or more of Al ³⁺ , Cr ³⁺ , Ga ³⁺ , In ³⁺ , Co ³⁺ , Fe ³⁺ and V ³⁺ , preferably Al ^{3 +} ;

B. Preparing a NaOH solution;

C. The mixed solution prepared in the step A is slowly added dropwise to the NaOH solution prepared in the step B under the condition of N ₂ protection, stirred, and the pH range of the mixed solution is adjusted to 8-10 by using the NaOH solution. Crystallization at 60 ° C -100 ° C for 12-48 hours, washed with water to neutral to obtain a hydrotalcite precursor;

D. The peptide is dissolved in water to obtain a force peptide solution, and the hydrotalcite precursor obtained by the step C is added to the force peptide solution, and the ion exchange is 0.5-24 in a water bath at 60-70 ° C under N ₂ protection. Hour, pick The mixture was washed with a deionized water (water temperature of 30-65 ° C) with a removal of C0 ₂ to neutrality, and dried at 50-70 ° C for 12-24 hours to obtain a force peptide intercalated hydrotalcite complex.

In the ion exchange process of the above step D, a microwave hydrothermal method is preferably employed to shorten the ion exchange time.

XRD, FT-IR, NMR and elemental analysis of the materials prepared above showed that the peptide was successfully inserted into the hydrotalcite layer; the /«-M XRD, In-situ IR, TG-DTA characterization showed the heat of the composite The stability has been significantly improved; the in vitro simulated release experiments show that the composite system can achieve a certain sustained release performance.

The invention has the advantages that the prepared force peptide intercalated hydrotalcite composite has better thermal stability under normal conditions due to utilizing the spatial confinement effect of hydrotalcite and the interaction between the host and the guest. A certain sustained release property is conducive to its storage and development of new routes of administration.

DRAWINGS

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is an X-ray powder diffraction (XRD) pattern of a force peptide intercalated hydrotalcite composite prepared in Example 1 of the present invention; the abscissa is 2 Θ, unit: degree; and the ordinate is intensity.

2 is a FT-IR spectrum of a force peptide intercalated hydrotalcite composite prepared in Example 1 of the present invention; the abscissa is a wave number, the unit is cm- ¹ ; the ordinate is a transmittance; wherein: a is a force peptide; b is a force peptide intercalated hydrotalcite complex.

Figure 3 is a NMR spectrum of the force peptide intercalated hydrotalcite composite prepared in Example 1 of the present invention; wherein: a is a force peptide; b is a force peptide intercalated hydrotalcite complex.

4 is an In-situ XRD spectrum of a force peptide intercalated hydrotalcite composite prepared in Example 1 of the present invention. Figure 5 is a graph showing the In-sUu IR spectrum of the force peptide intercalated hydrotalcite composite prepared in Example 1 of the present invention. 6 is a thermogravimetric (TG) curve of a force peptide intercalated hydrotalcite composite prepared in Example 1 of the present invention; wherein: a is a force peptide; b is a force peptide intercalated hydrotalcite complex.

Figure 7 is a differential thermal (DTA) curve of the force peptide intercalated hydrotalcite composite prepared in Example 1 of the present invention; wherein: a is a force peptide; b is a force peptide intercalated hydrotalcite complex. detailed description

[Embodiment 1]

Step A: 3.846 g (0.015 mol) of solid M _g (N0 ₃ ) ₂ * 63⁄40 and 2.8135 _g (0.0075 mol) of solid Α 1 (Ν 0 ₃ ) ₃ · 93⁄40 dissolved in 50 mL of deionized water with C0 ₂ removed (Solution I); further 4g (0.018mol) of force peptide and 2.5g (0.0625mol) of NaOH dissolved in 50mL of deionized water (Case II) with CO ₂ removed;

Step B: The solution II was placed in a four-necked flask, and while stirring under a condition of N ₂ gas, the solution I was slowly added dropwise to the solution II while stirring, and the mixture was dropped for about 0.5 h. After the completion of the dropwise addition, the pH was adjusted to 8 with a 5 mol/L NaOH solution;

Step C: 60 ° C water bath reaction for 24 h;

Step D: After centrifugation with deionized hot water except C0 ₂ to pH 7, and dried at 70 ° C for 12 h, a magnesium-aluminum-type force peptide intercalated hydrotalcite composite was obtained.

An X-ray powder diffraction (XRD) pattern of the force peptide and the force peptide intercalated hydrotalcite composite prepared in Example 1 was obtained using an XRD-6000 X-ray diffractometer of Shimadzu Corporation, as shown in Fig. 1.

The FT-IR spectrum of the force peptide and the force peptide intercalated hydrotalcite complex prepared in Example 1 was obtained using a Nicolet 560 type Fourier infrared spectrometer, as shown in Fig. 2.

The NMR spectrum of the force peptide and the force peptide intercalated hydrotalcite complex prepared in Example 1 was obtained using a nuclear magnetic apparatus of the AV300 type of Bmcker, Germany, as shown in Fig. 3.

Using the X-ray diffractometer of the Dutch PANalytical X' Pert PRO MPD type, The In-situ XRD spectrum of the force peptide intercalated hydrotalcite complex prepared in Example 1 is shown in FIG. The In-sUu IR spectrum of the force peptide intercalated hydrotalcite composite prepared in Example 1 was obtained using a Fourier infrared spectrometer of Model 560 from Nicolet, as shown in FIG.

The thermogravimetric (TG) curve of the force peptide and the hydropeptide intercalated hydrotalcite complex prepared in Example 1 was obtained using a HCT-2 type microcomputer differential thermostat from Beijing Hengjiu Scientific Instrument Factory, as shown in Fig. 6. Based on the thermogravimetric curve as shown in Figure 6, the differential thermal (DTA) curve of the force peptide and the force peptide intercalated hydrotalcite complex was obtained.

It can be seen from the XRD spectrum, the FT-IR spectrum and the NMR spectrum that the obtained anion of the hydrotalcite layer is a force peptide anion.

From the In-situ XRD spectrum, the In-situ IR spectrum and the TG-DTA curve, the thermal stability of the force peptide intercalated hydrotalcite complex was significantly improved compared with the force peptide.

[Embodiment 2]

The slurry obtained in the step B of Example 1 was transferred to a microwave rapid digestion tank, and the temperature was controlled by microwave heating at 90 ° C for 0.5 h, and the obtained product was centrifugally washed with deionized hot water from which C0 ₂ was removed until the pH was about 7, dried at 70 ° C for 12 h.

[Embodiment 3]

Step A: Weigh 15.384g Mg(N0 ₃ ) ₂ · 63⁄40 and 11.2539g Α1(Ν0 ₃ ) ₃ · 93⁄40 dissolved in 100ml to C0 ₂ , deionized water to prepare mixed salt solution i, and another 7.2 _g NaOH dissolved 100 ml of deionized water (alkaline solution ii) of C0 ₂ was removed and placed in a 500 mL four-necked flask;

Step B: Slowly drip the salt solution i into the alkali solution mash under a water bath of 70 ° C, N ₂ protection, and vigorous stirring, adjust the pH to about 10.0, and react for 40 hours; Step C: The product is washed and filtered to a pH of 7; a large amount of fresh filter cake is reserved for ion exchange, and only a small amount of sample is taken out and dried at 70 ° C for 18 hours to obtain N0 ₃ -Mg ₂ Al-LDHs, Mg ²⁺ /Al ³⁺ =2;

Step D: The appropriate amount of peptide was dissolved in 200 mL of deionized water with CO ₂ removed and placed in a 500 mL four-necked flask, and an appropriate amount of the filter cake obtained by the step C was added thereto to adjust the pH to 9;

Step E: Ion exchange for 24 h under N ₂ protection, vigorous stirring, temperature control at 60 ° C, the obtained product was washed and centrifuged 4 times with deionized water with C0 ₂ removed, and the precipitate was dried in a vacuum drying oven. A layered composite material containing an interlayer of a force peptide anion can be obtained.

[Embodiment 4]

The mixed solution obtained in the step D of Example 3 was transferred to a microwave rapid digestion tank, and the temperature was controlled by microwave heating at 90 ° C for 0.5 h, and the obtained product was centrifugally washed with a deionized hot water with C0 ₂ removed to a pH of about Dry at 7, 70 ° C for 12-24h.

[Embodiment 5]

Step A: The mixed salt solution i in the step A of Example 3 was quickly added to the alkali solution crucible under high-speed stirring, and after stirring for 20 minutes, the obtained product was washed and centrifuged with deionized water from which C0 ₂ was removed. 4 times;

Step B: taking an appropriate amount of the precipitate obtained in step A and an appropriate amount of the force peptide dissolved in deionized water with CO ₂ removed, adjusting the pH to 8;

Step C: Transfer the solution obtained in step B to an autoclave, and react at a constant temperature of 90 ° C for 16 h; Step D: The product was washed and centrifuged 4 times with deionized water with C0 ₂ removed, and the precipitate was dried in a vacuum at room temperature to obtain a layered composite material having an interlayer-containing peptide anion.

It can be seen from the XRD spectrum, the FT-IR spectrum, and the NMR spectrum that the obtained anion of the hydrotalcite layer is a force peptide anion.

Claims

Claim

A force peptide intercalated hydrotalcite composite characterized in that the chemical formula of the force peptide intercalated hydrotalcite compound is: [M ² Y _x M ³⁺ _x (OH) ₂ f ⁺ (A ) _x * M3⁄40, wherein M ²⁺ represents any one or more of divalent metal ions Mg ²⁺ , Co ²⁺ , Ni ²⁺ , Ca ²⁺ , Cu ²⁺ , Fe ²⁺ and Mn ²⁺ ; M ^{3 +} represents any one or more of the trivalent metal ions Al ³⁺ , Cr ³⁺ , Ga ³⁺ , In ³⁺ , Co ³⁺ , Fe ³⁺ and V ³⁺ ; A—as the hydrotalcite layer Anionic force peptide anion; X ranges from 0.2 to 0.33, and m ranges from 0 to 2.

2. The force peptide intercalated hydrotalcite composite according to claim 1, wherein M ²⁺ is N ₁ ²⁺ ,

Co ²⁺ or Mg ²⁺ , M ³⁺ is Al ³⁺ .

3. A method of preparing a force peptide intercalated hydrotalcite composite according to claim 1, the method comprising the steps of:

a preparation of a mixed solution of a soluble divalent metal salt and a soluble trivalent metal salt, wherein the concentration of the divalent metal ion is 0.01-1.0 M, and the molar ratio of the divalent metal ion to the trivalent metal ion is 2-4, divalent The metal ion is any one or more of Mg ²⁺ , Co ²⁺ , Ni ²⁺ , Ca ²⁺ , Cu ²⁺ , Fe ²⁺ and Mn ²⁺ , and the trivalent metal ion is Al ³⁺ , Cr ³ Any one or more of ⁺ , Ga ³⁺ , In ³⁺ , Co ³⁺ , Fe ^{3+ ,} and V ³⁺ ;

b. formulating a mixed solution of a force peptide and NaOH;

c The 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 of the mixed solution is adjusted to 7-10 by using NaOH solution, at 60 Crystallization at °C-70 °C for 2-24 hours, washing with water to neutrality, drying at 50-70 °C for 12-24 hours, to obtain a force peptide intercalated hydrotalcite complex.

The method according to claim 3, wherein the divalent metal ion is Mg ²⁺ , Co ²⁺ or M ²⁺ , and the trivalent metal ion is Al ³⁺ .

The method according to claim 3, wherein in the crystallization of the step c, a method of microwave water heat is employed.

6. The method for preparing a polypeptide drug intercalated hydrotalcite according to claim 1, wherein the method comprises the following steps:

(1) preparing a mixed solution of a soluble divalent metal salt and a soluble trivalent metal salt, wherein the concentration of the divalent metal ion is 0.01-1.0 M, and the molar ratio of the divalent metal ion to the trivalent metal ion is 2-4, The valence metal ion is any one or more of Mg ²⁺ , Co ²⁺ , Ni ²⁺ , Ca ²⁺ , Cu ²⁺ , Fe ²⁺ and Mn ²⁺ , and the trivalent metal ion is Al ³⁺ , Cr Any one or more of ³⁺ , Ga ³⁺ , In ³⁺ , Co ³⁺ , Fe ^{3+ ,} and V ³⁺ ;

(2) preparing a NaOH solution;

(3) The mixed solution prepared in the step (1) is slowly added dropwise to the NaOH solution prepared in the step (2) under the condition of N ₂ protection, stirred, and the pH of the mixed solution is adjusted to be adjusted by using the NaOH solution to 8-10, crystallization at 60 ° C -100 ° C for 12-48 hours, washed with water to neutral to obtain a hydrotalcite precursor;

(4) The force-taking peptide is dissolved in water to obtain a force peptide solution, and the hydrotalcite precursor obtained by the step (3) is added to the force peptide solution, and the ion exchange is carried out under the condition of 60-70 ° C water bath and N ₂ protection. 0.5-24 hours, washed by water to neutral, dried at 50-70 °C for 12-24 hours, and the compound peptide intercalated hydrotalcite is obtained.

The method according to claim 6, wherein the divalent metal ion is Mg ²⁺ , Co ^: or M ²⁺ , and the trivalent metal ion is Al ³⁺ . Use microwave water heat method.