TW201402113A - Indirubin particle and manufacturing method thereof - Google Patents

Abstract

The present invention relates to an indirubin particle and a manufacturing method thereof. The indirubin particle comprises: a cluster constituted by a plurality of indirubin molecules and a plurality of polymeric ligands covering a surface of the cluster. The polymeric ligands provide functionality of hydrophilicity to allow the indirubin molecules to sufficiently spread in an aqueous phase system thereby improving the applicability of indirubin in a medicine conveyance system. Further, the manufacturing method of the present invention belongs to physical property modification treatment and is a simple process for improving the drawbacks of the conventional manufacturing processes that are complicated, have low productivity, and have difficult in separation.

Description

Qiyuhong granule and preparation method thereof

The present invention relates to an organic drug coated with a polymer, and more particularly to an indigo red particle having a polymer ligand and a preparation method thereof.

Indirubin is a kind of bis-indole molecule, which is widely used in biomedical fields or medical beauty products. Among them, since indirubin has a therapeutic effect on chronic diseases, it has been conventionally used as a clinical therapeutic drug for cancer, such as leukemia. For example, in Non-Patent Document 1, it is proposed that a derivative of indirubin has antitumor effect. Features. In addition, based on the function of promoting cell activity, indirubin is also recently applied to one of the compound components of hair growth.

At present, the method of obtaining indirubin is generally obtained by extraction from natural plants (Daoqingye), by chemical synthesis or from commercial sources, but because the indirubin is a hydrophobic molecule, it leads to water solubility. Poorly, for example, in Non-Patent Document 2, it is revealed that the solubility of the indirubin molecule in water is less than 1 mg/L; and because the drug delivery system almost uses water as a solvent to provide a medium for drug delivery. Therefore, if only the indirubin is added to the water, it is difficult to be dissolved, and the indirubin molecule cannot be sufficiently diffused in the medium, so that the indirubin molecule cannot pass the aqueous phase system for drug delivery.

Based on the above disadvantages, conventional techniques have revealed a modified treatment of indirubin derivatives to enhance hydrophilicity, for example, in non-patent literature. In the third, the indigo red derivative and its preparation method are proposed, which adopts a polar functional group to carry out a substitution reaction on the indirubin molecule to chemically modify the indirubin molecule, thereby obtaining a indirubin derivative. .

Although the solubility of the indirubin derivative obtained by the above chemical method in the aqueous phase may be higher than that of the unmodified indomethacin molecule, the reaction step performed by such a chemical modification method is complicated and difficult. Effective control of the parameters of the reaction, resulting in the cost of the process time and cost; at the same time, due to the poor selectivity of the reaction, the product obtained after the completion of the reaction also contains by-products, thus resulting in a decrease in the yield of the indirubin derivative. Furthermore, it is necessary to further separate the products of similar properties to obtain the desired indirubin derivative, which further limits the utilization of indirubin in the industry.

Non-Patent Document 1: Soo-A Kim et al., Clin. Cancer Res. 2007, 13, 253-259

Non-Patent Document 2: Rolf Jautelat et al., ChemBioChem 2005, 6, 531-540

Non-Patent Document 3: Konstantina VOUGOGIANNOPOULOU et al., J Med Chem. 2008 October 23; 51(20): 6421-6431

In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to disclose a physical modification treatment of indirubin to improve the hydrophilic property, reduce the complicated process of the reforming process, and cause time and cost, and solve the problem due to chemistry. The disadvantages of low yield due to synthesis and difficulty in separating the product.

In order to achieve the above object, the present invention provides a indirubin red particle comprising: a cluster composed of a plurality of indirubin molecules; and a plurality of polymer ligands, Covering the surface of the cluster, each of the polymer ligands being a surfactant, and the polymer ligands comprise a first ligand and a second ligand, each of the first ligands being a tri-copolymer (triblock copolymer), and each of the second ligand systems is a stabilizer and is intertwined with the first ligand.

In an embodiment, the first ligand is a polyglycol ether copolymer (poloxamer) and the second ligand is selected from the group consisting of polyethylene glycol (PEG) and water-soluble vitamin E (TPGS, tocopheryl polyethylene glycol succinate). One or both.

Further, the particle size of the indirubin particles is preferably from about 50 nm to about 5000 nm.

Furthermore, the present invention further provides a method for preparing indirubin red particles, the method comprising the steps of: (a) dissolving an indirubin molecule and a tri-embedded copolymer in an organic solvent to form an organic solution, wherein the three inlays The copolymer is a surfactant; (b) mixing the organic solution with an aqueous solution containing a stabilizer to form a mixed solution, wherein the stabilizer is a surfactant; and (c) stirring the mixed solution The eucalyptus red granules were obtained by emulsifying the mixed solution.

In addition, in the embodiment, when the step (c) is performed, the mixing method of the mixed solution can be directly stirred by the instrument or transmitted through the ultrasonic wave. Stir.

In addition, the present invention further provides a method for preparing indirubin red particles, the method comprising the steps of: (a) dissolving an indirubin molecule and a tri-embedded copolymer in an organic solvent to form an organic solution, wherein the three inlaid The copolymer is a surfactant; (b) the organic solution is freeze-dried to remove the organic solvent to form a precursor powder; and (c) the precursor powder is mixed with an aqueous solution containing a stabilizer to obtain Indirubin particles, wherein the stabilizer is a surfactant.

In the embodiment of each of the above preparation methods, when the step (a) is carried out, the content of the indirubin molecule is 0.5 to 20 parts by weight, and the content of the tri-embedded copolymer is 50 parts by weight.

In summary, the preparation method of the indirubin red particles disclosed in the present invention is a physical modification treatment method for indirubin, and the steps of the preparation method are simple, thereby effectively reducing the time and cost of the process, and at the same time, Since the coating configuration in each of the preparation methods involves only intermolecular forces, the yield and separation problems derived from the chemical synthesis reaction procedure are avoided. Furthermore, the indirubin particles obtained by each of the preparation methods have good hydrophilicity, so that the indirubin red particles are stably dispersed in the aqueous phase system, and then the indigo red is transferred during drug delivery. Fully spread in the medium (water) for transport, thereby expanding the applicability of indirubin in the field of biomedicine.

To fully clarify the objects, features, and advantages of the present invention, those of ordinary skill in the art of the invention can understand the invention and practice the invention. Schematic, a detailed description of the present invention, illustrated as follows:

definition

In this document, "a" is used to describe the elements and components of the invention. This usage is provided for convenience only, and is intended to be inclusive of the generality of the invention.

Where a quantity, a concentration, or other value or parameter is expressed by a range, a preferred range, or a series of upper and lower limits, it is understood to be a specific disclosure of the upper or lower value of any range or the lower limit of any range or All ranges of preferred values are constructed regardless of whether such ranges are disclosed separately. In addition, when a range of values is recited herein, unless otherwise stated, the range shall include its endpoints and all integers and fractions within the range.

In the present invention, before the object can be attained, the numerical value should be understood as having the accuracy of the number of significant digits. For example, the number 40 should be understood to cover the range from 35.0 to 44.9, while the number 40.0 should be understood to cover the range from 39.50 to 40.49.

The indigo red granule of the invention

Referring to FIG. 1, the present invention discloses an indirubin red particle 1 comprising a cluster 11 and a plurality of polymer ligands 12. The indirubin particles 1 exhibit a dispersion in the aqueous phase system.

The cluster 11 is composed of a plurality of indirubin molecules 111, wherein The indirubin used may be obtained by natural plant extraction, chemical synthesis or from commercial sources. In the embodiment of the present invention, the source of indirubin is the extract of Daqingye (purchased from Shanchuan Biotechnology, the purity is about 98%) or chemically synthesized indirubin red molecules (purity higher than 99%).

The polymer ligands 12 coat the surface of the cluster 11 in a dynamic equilibrium manner in a liquid environment, and each of the polymer ligands 12 is a molecule having a hydrophilic functional group and a hydrophobic functional group, that is, a surfactant, and the polymer ligand 12 includes a plurality of first ligands 121 and a plurality of second ligands 122, wherein each of the first ligands 121 is a triple-embedded copolymer, and the three intercalation copolymerization The article has three long chains linked by covalent bonds, and each of the long chains is a homopolymer, and the nonionic triad copolymer (PEO-PPO) used as the first ligand 121 in the present invention -PEO), the central long chain 1211 is hydrophobic, and the long chain 1212 on both sides thereof is hydrophilic, and therefore, when the first ligand 121 coats the cluster 11 in water, each of the first ligands The 121 system adsorbs the cluster 11 by the central hydrophobic long chain 1211, and guides and disperses the cluster 11 into the water by using the hydrophilic long chain 1212 on both sides and the water molecule attraction; each of the second ligands 122 Is a stabilizer and is intertwined with each of the first ligands 121 to collectively coat the cluster 11, and the function of the second ligand 122 is The coating configuration of the first ligand 121 in the dynamic balance avoids the collapse of the indigo red particle 1 due to the voiding of the coating configuration, thereby stabilizing the coating configuration of the indirubin red particle 1 in the aqueous solution, and The second ligand 122 also inhibits the aggregation of the indirubin particles 1 by their hydrophilic functional groups, thereby preventing the precipitation of the indirubin particles 1 and allowing them to have good dispersibility in an aqueous solution.

It should be understood that the polymer ligand 12 of the present invention may also select a biocompatible polymer as a source to enable the indigo red particle 1 of the present invention to be exhibited in the field of biomedical or medical products. Good efficacy, for example, the source may be a polyglycol ether copolymer, polyethylene glycol or water-soluble vitamin E, etc., but is not limited thereto, and those having ordinary knowledge in the art to which the present invention pertains should understand that Other polymers which are characteristic of the ligand 12 and which are biocompatible can also serve as a source of the polymer ligand 12 of the present invention if it provides effective dispersion or stabilizing function.

Furthermore, since the size of the drug particles is one of the important factors determining whether the general drug particles can be bioabsorbed after being released, and the particle size which can be absorbed by the human body (for example, skin) or in the human body (for example, the gastrointestinal tract) The particle size of the indirubin particles 1 of the present invention is preferably between about 50 and 5000 nm, and if the particle diameter is greater than 5000 nm, the crucible is caused. The absorption of the jade red particles 1 is difficult, and at the same time, the jadeite particles 1 in the water form a colloidal precipitate to cause sedimentation.

The first embodiment of the preparation method of the invention: preparing the indigo red particles by emulsification and solvent evaporation method

In the first embodiment, the indirubin particles are prepared by emulsification and solvent evaporation.

First, the indirubin molecule and the tri-embedded copolymer are dissolved in an organic solvent to form an organic solution, and the ratio of the indirubin to the tri-embedded compound is between 1:100 and 2:5 (ie, 靛The content of the jade red molecule is 0.5 to 20 parts by weight, and the content of the tri-embedded copolymer is 50 parts by weight), the three-in-line type The copolymer is commercially available from Pluronic L81, Pluronic F68 or Pluronic F127 (with PEO-PPO-PEO structure), and the organic solvent is dimethyl sulfoxide, acetone, ethanol or methanol. Among them, acetone is preferably used as the organic solvent. It should be understood that depending on the type or weight of the organic solvent used, the content of indirubin should also be adjusted according to the solubility or the amount of dissolution. For example, when the organic solvent uses 0.5 to 5 mL of acetone, the jade is used. The red content is 1~10 mg.

Next, 100 mL of an aqueous solution containing a stabilizer is provided, and the organic solvent is mixed with the aqueous solution to form a mixed solution, wherein the stabilizer is selected from one or both of polyethylene glycol or water-soluble vitamin E. The water-soluble vitamin E is composed of vitamin E and polyethylene glycol. Preferably, polyethylene glycol is used as a stabilizer.

Finally, the mixed solution is stirred to bring the mixed solution to an emulsified state, and then, the mixed solution is heated (to 150 ° C) or the mixed solution is directly placed at room temperature to volatilize the organic solvent therein, that is, The indirubin red particles of the present invention which are colloidally dispersed are obtained in the remaining aqueous phase solution. In the method of stirring, the apparatus can be directly stirred by means of an instrument (such as a magnet stirrer and a blender) or a machine (such as a stirrer with a blade stirrer), or the mixture can be stirred by ultrasonic vibration. .

The particle size distribution range of the indirubin red particles obtained by the preparation method is mainly concentrated in the range of 400 to 2000 nm.

A second embodiment of the preparation method of the present invention: preparing ruthenium red granules by freeze-drying method

In the second embodiment, it is prepared by freeze drying. 靛玉红粒.

First, the indirubin molecule and the tri-embedded copolymer are dissolved in an organic solvent to form an organic solution, and the ratio of the indirubin to the tri-embedded compound is between 1:100 and 2:5. The intercalated copolymer is commercially available from Pluronic L81, Pluronic F68 or Pluronic F127, and the organic solvent is dimethyl hydrazine, acetone, ethanol or methanol, and among them, dimethyl hydrazine is preferably used as the organic solvent. It should be understood that depending on the type or weight of the organic solvent used, the content of indirubin should also be adjusted depending on the solubility or the amount of dissolution. For example, when 2 mL of dimethyl hydrazine is used as the organic solvent, 靛The content of jade red can be 0.5~20 mg.

Next, the organic solution is freeze-dried for about 48 to 98 hours. During the treatment, the organic solution is first placed in liquid nitrogen to a low temperature environment to reach below the eutectic point of the organic solution to freeze to form a The cured product is then sent to a collector of a freeze dryer that has been pre-cooled to -40 ° C to -48 ° C, and the vacuum pump is turned on to reduce the pressure in the collector to 5 to 15 Pa by The low-pressure environment sublimes the organic solvent component in the cured product, and the precursor powder is formed after the organic solvent component is removed.

Finally, 100 mL of an aqueous solution containing a stabilizer is provided, wherein the stabilizer is selected from one or both of polyethylene glycol or water-soluble vitamin E, and then the precursor powder is mixed with the aqueous solution to obtain a gel. The indigo red particles of the present invention are dispersed.

The particle size distribution range of the indirubin red particles obtained by the preparation method is mainly concentrated in the range of 60 to 1200 nm.

Example

For the sake of brevity and convenience of description, the embodiments obtained by the above-mentioned preparation method are shown in Table 1 below, and the technical means of the present invention are more specifically disclosed by the embodiments to prove that the present invention can be implemented. However, the invention is not limited to the following embodiments.

The abbreviations in Table 1 are as follows: Size: Average particle size of indirubin red particles

PDI: polydispersity index

I: 靛玉红分子

PF68: Pluronic F68

PEG600: polyethylene glycol

TPGS: Water-soluble vitamin E

DMSO: dimethyl hydrazine

Preparation method 1: Preparation method of the first embodiment

Preparation method 2: Preparation method of second embodiment

Among them, the methods for removing the organic solvent in the preparation methods of Examples 1 to 4 are: volatilization at room temperature, volatilization at 120 ° C and volatilization at 150 ° C; volatilization at room temperature means placing the mixed solution at room temperature to make The organic solvent is naturally volatilized; the volatilization at 120 ° C means heating the mixed solution to 120 ° C to volatilize the organic solvent; and volatilizing at 150 ° C means heating the mixed solution to 150 ° C to volatilize the organic solvent.

Based on the steps disclosed in the first embodiment and the second embodiment, since the organic solvent placed in the process is removed at the end of the preparation method, the prepared indirubin particle system is present at 100. In the aqueous phase system of mL, it can be known from the content of the components disclosed in Table 1 that the molecular content of the indirubin in the examples is between 2.5 mg and 20 mg, so that the indigo red particles of the present invention can be formed in water. The concentration is 2.5 mg of indirubin molecule / 100 mL to 20 mg of indirubin molecule / 100 mL of colloidal dispersion, and after completion of the preparation method, the observation system found no precipitation of each example, and its attribution The indirubin particles of the present invention have good hydrophilicity.

In order to show the morphology of indirubin red particles in water, please refer to Fig. 2, which is a TEM photograph of a sample taken from an aqueous solution containing indirubin particles, which is based on an aqueous solution containing indirubin particles. Example 9 and scale up 150 times the product obtained, the number found in the figure The indirubin red particles (light gray particles) with a particle size of about 200-300 nm are more sufficient to prove that the indirubin red particle system of the embodiment does exist and is dispersed in water instead of precipitation, so it is concluded that the present invention The indirubin particles have good hydrophilicity, which in turn provides a stable dispersion in the aqueous phase system.

The invention has been described above in terms of the preferred embodiments, and it should be understood by those skilled in the art that the present invention is not intended to limit the scope of the invention. It should be noted that variations and permutations equivalent to those of the embodiments are intended to be included within the scope of the present invention. Therefore, the scope of protection of the present invention is defined by the scope of the patent application.

1‧‧‧靛玉红粒

11‧‧‧ cluster

111‧‧‧靛玉红分子

12‧‧‧ polymer ligands

121‧‧‧First ligand

1211‧‧ long chain

1212‧‧ long chain

122‧‧‧Second ligand

Figure 1 is a schematic view showing the structure of the indirubin red particles of the present invention.

Figure 2 is a TEM photograph of a sample taken from an aqueous solution containing indirubin particles.

1‧‧‧靛玉红粒

11‧‧‧ cluster

111‧‧‧靛玉红分子

12‧‧‧ polymer ligands

121‧‧‧First ligand

1211‧‧ long chain

1212‧‧ long chain

122‧‧‧Second ligand

Claims (14)

An indirubin red particle comprising: a cluster consisting of a plurality of indirubin molecules; and a plurality of polymer ligands covering the surface of the cluster, each The polymer ligand is a surfactant, and the polymer ligands comprise a first ligand and a second ligand, each of the first ligands being a triblock copolymer, and each of the second The coordination system is a stabilizer and is intertwined with the first ligand. The indirubin particles according to claim 1, wherein the first ligand is a polyglycol ether copolymer (POLoxamer), and the second ligand system is selected from the group consisting of polyethylene glycol (PEG) and water-soluble vitamins. Group of E (TPGS, tocopheryl polyethylene glycol succinate). The indirubin particles according to claim 2, wherein the second ligand is polyethylene glycol. The indirubin red particles according to any one of claims 1 to 3, wherein the indirubin red particles have a particle diameter of about 50 nm to 5000 nm. The indirubin particles according to claim 4, wherein the indigo red particles have a particle diameter of about 364.8 nm to 1603 nm. A method for preparing indirubin red particles, the method comprising the steps of: (a) dissolving an indirubin molecule and a tri-embedded copolymer in an organic solvent to form an organic solution, wherein the tri-embedded copolymer is an interfacial activity (b) mixing the organic solution with an aqueous solution containing a stabilizer to form a a mixed solution in which the stabilizer is a surfactant; and (c) stirring the mixed solution to emulsify the mixed solution to obtain indirubin particles. The preparation method according to claim 6, wherein in the step (a), the content of the indirubin molecule is 0.5 to 20 parts by weight, and the content of the tri-embedded copolymer is 50 parts by weight. The preparation method according to claim 6 or 7, wherein in the step (c), the mixing solution is stirred by directly stirring the device or by ultrasonic vibration. The preparation method according to claim 6, wherein the tri-embedded copolymer is a polyglycol ether copolymer, and the stabilizer is selected from the group consisting of polyethylene glycol and water-soluble vitamin E. The preparation method of claim 9, wherein the stabilizer is polyethylene glycol. A method for preparing indirubin red particles, the method comprising the steps of: (a) dissolving an indirubin molecule and a tri-embedded copolymer in an organic solvent to form an organic solution, wherein the tri-embedded copolymer is an interfacial activity (b) lyophilizing the organic solution to remove the organic solvent to form a precursor powder; and (c) mixing the precursor powder with an aqueous solution containing a stabilizer to obtain indirubin particles, wherein The stabilizer is a surfactant. The preparation method according to claim 11, wherein in step (a), the content of the indirubin molecule is from 0.5 to 20 parts by weight, and the three-inlaid copolymer The content is 50 parts by weight. The preparation method according to claim 11, wherein the tri-embedded copolymer is a polyglycol ether copolymer, and the stabilizer is selected from the group consisting of polyethylene glycol and water-soluble vitamin E. The preparation method of claim 13, wherein the stabilizer is polyethylene glycol.