CN113350525A - Porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound and preparation method and application thereof - Google Patents
Porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, a preparation method and application thereof, and belongs to the technical field of medicines. The porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound provided by the invention comprises porous starch and artemisinin hydroxypropyl-beta-cyclodextrin inclusion compounds loaded in pores of the porous starch, wherein the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound comprises artemisinin and hydroxypropyl-beta-cyclodextrin coated on the surface of the artemisinin. The invention takes fat-soluble artemisinin as an effective component, hydroxypropyl-beta-cyclodextrin as an inclusion material and porous starch as a carrier, can remarkably improve the water solubility of the artemisinin, and further improves the bioavailability and antimalarial activity of the artemisinin; the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound provided by the invention has good biological safety, and can avoid potential hazards and toxic and side effects possibly existing in the existing artemisinin derivatives.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, and a preparation method and application thereof.
Background
The artemisinin is used as an effective medicinal component extracted and separated from medicinal plant artemisia apiacea, and shows lower toxic and side effects and better antimalarial effect in cells and animal models. In antimalarial and therapeutic experiments, oral artemisinin administration or intravenous administration is mainly adopted, but artemisinin is poor in water solubility and low in bioavailability and is difficult to be sufficiently absorbed and utilized in a human body. Thus, large doses are often required to achieve equivalent therapeutic effects, but the effect is not ideal.
The main approach for increasing the water solubility of artemisinin is to structurally modify functional groups of artemisinin and synthesize a series of glycosylated or basic derivatives, such as artemether, dihydroartemisinin or artesunate. Although the water solubility of the artemisinin is increased to a certain extent, the action mechanism of the artemisinin on the aspects of reproductive toxicity and genetic toxicity of human bodies is not clear enough, and potential safety hazards exist.
Disclosure of Invention
The invention aims to provide a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, a preparation method and application thereof.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, which comprises porous starch and an artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound loaded in pores of the porous starch, wherein the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound comprises artemisinin and hydroxypropyl-beta-cyclodextrin coated on the surface of the artemisinin.
Preferably, the content of the artemisinin in the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound is 10-30%, and the content of the hydroxypropyl-beta-cyclodextrin is 10-30%.
The invention provides a preparation method of a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, which comprises the following steps:
performing inclusion treatment on a first mixed system containing artemisinin and hydroxypropyl-beta-cyclodextrin to obtain an artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound;
and (3) adsorbing a second mixed system containing the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound and porous starch to obtain the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound.
Preferably, the mass ratio of the hydroxypropyl-beta-cyclodextrin to the artemisinin is (1.8-2.2): 1.
preferably, the preparation method of the first mixed system comprises: mixing the artemisinin solution with the hydroxypropyl-beta-cyclodextrin solution to obtain a first mixed system; the mass content of the artemisinin in the artemisinin solution is 4.5-85%, and the mass content of the hydroxypropyl-beta-cyclodextrin in the hydroxypropyl-beta-cyclodextrin solution is 4-85%.
Preferably, the temperature of the inclusion treatment is 40-60 ℃ and the time is 1-6 h.
Preferably, the mass ratio of the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound to the porous starch is 1: (0.8 to 1.2).
Preferably, the preparation method of the second mixed system comprises the following steps: mixing the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound with the porous starch dispersion system to obtain a second mixed system; the mass content of the porous starch in the porous starch dispersion system is 30-70%.
Preferably, the temperature of the adsorption treatment is 30-40 ℃, and the time is 0.5-2 h.
The invention provides application of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in the technical scheme or the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound prepared by the preparation method in the technical scheme in preparation of a drug for treating malaria.
The invention provides a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, which comprises porous starch and an artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound loaded in pores of the porous starch, wherein the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound comprises artemisinin and hydroxypropyl-beta-cyclodextrin coated on the surface of the artemisinin. The invention takes fat-soluble artemisinin as an effective component, hydroxypropyl-beta-cyclodextrin as an inclusion material and porous starch as a carrier, can obviously improve the water solubility of the artemisinin, and further improves the bioavailability and antimalarial activity of the artemisinin. The porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound provided by the invention is used as an active ingredient to prepare an antimalarial drug, so that the problem of large dosage of oral administration, injection and the like of the existing artemisinin can be solved, the interval time of medication is prolonged, and the artemisinin can be better absorbed and metabolized in vivo. Meanwhile, compared with the existing artemisinin derivative, the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound provided by the invention has good biological safety, and can avoid potential hazards and toxic and side effects possibly existing in the existing artemisinin derivative. The results of the examples show that the solubility and bioavailability of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS) and the concentration of the intracerebral drug are remarkably improved compared with artemisinin bulk drugs, dihydroartemisinin and commercial artemisinin piperaquine tablets.
The invention provides a preparation method of a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, which comprises the following steps: performing inclusion treatment on a first mixed system containing artemisinin and hydroxypropyl-beta-cyclodextrin to obtain an artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound; and (3) adsorbing a second mixed system containing the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound and porous starch to obtain the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound. The method provided by the invention is simple in process flow and easy for industrial production.
Drawings
FIG. 1 is an SEM image of artemisinin (a), hydroxypropyl-beta-cyclodextrin (b), artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (c), porous starch (d) and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (e, f);
FIG. 2 is an XRD pattern of artemisinin (A), hydroxypropyl-beta-cyclodextrin (B), porous starch (C) and porous starch loaded hydroxypropyl-beta-cyclodextrin inclusion compound (D);
FIG. 3 is a graph showing the dissolution curves of porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (A), dihydroartemisinin (B), artemisinin piperaquine tablet (C) and artemisinin original drug (D) in water;
FIG. 4 is a dissolution curve diagram of porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (A), dihydroartemisinin (B), artemisinin piperaquine tablet (C) and artemisinin original drug (D) in artificial gastric juice;
FIG. 5 is a dissolution curve diagram of porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (A), dihydroartemisinin (B), artemisinin piperaquine tablet (C) and artemisinin original drug (D) in an artificial intestinal juice;
FIG. 6 is a graph showing the results of porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS), Dihydroartemisinin (DHA), artemisinin piperaquine tablets and artemisinin original drug (ART) on the infection rate of CM rats;
FIG. 7 is a graph showing the bioavailability results of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound fed to rats;
FIG. 8 is a graph showing the comparison of the artemisinin concentration in each tissue after feeding rats with artemisinin original drug (ART), Dihydroartemisinin (DHA), artemisinin piperaquine tablets and porous starch loaded hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS) for 0.5 h;
FIG. 9 is a graph showing the comparison of the artemisinin concentration in each tissue after feeding rats lh with artemisinin original drug (ART), Dihydroartemisinin (DHA), artemisinin piperaquine tablet and porous starch loaded with artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS).
FIG. 10 is a graph showing the comparison of the artemisinin concentration in each tissue after feeding rats with artemisinin original drug (ART), Dihydroartemisinin (DHA), artemisinin piperaquine tablet and porous starch loaded hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS) for 2 h.
FIG. 11 is a graph showing the comparison of the artemisinin concentration in each tissue after feeding rats with artemisinin original drug (ART), Dihydroartemisinin (DHA), artemisinin piperaquine tablet and porous starch loaded hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS) for 4 h.
FIG. 12 is a graph showing the comparison of the artemisinin concentration in each tissue after feeding rats with artemisinin original drug (ART), Dihydroartemisinin (DHA), artemisinin piperaquine tablet and porous starch loaded hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS) for 6 h.
FIG. 13 is a graph showing the comparison of the artemisinin concentration in each tissue of rats fed with artemisinin original drug (ART), Dihydroartemisinin (DHA), artemisinin piperaquine tablets and porous starch loaded hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS) for 8 h.
FIG. 14 is a graph showing the comparison of the artemisinin concentration in each tissue after feeding rats with artemisinin original drug (ART), Dihydroartemisinin (DHA), artemisinin piperaquine tablet and porous starch loaded hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS) for 12 h.
FIG. 15 is a graph showing the comparison of the artemisinin concentration in each tissue after feeding rats with artemisinin original drug (ART), Dihydroartemisinin (DHA), artemisinin piperaquine tablet and porous starch loaded hydroxypropyl-beta-cyclodextrin inclusion compound (AHPS) for 24 h.
FIG. 16 is a graph showing the bioavailability results of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound fed to rats in the heart;
FIG. 17 is a graph showing the results of bioavailability in the liver of rats fed with artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound;
FIG. 18 is a graph showing the results of bioavailability in the spleen of rats fed a combination of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl- β -cyclodextrin inclusion;
FIG. 19 is a graph showing the bioavailability results of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound fed to rats in the lungs;
FIG. 20 is a graph showing the bioavailability results in the kidney of rats fed a combination of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl- β -cyclodextrin inclusion;
figure 21 is a graph of bioavailability results in the brain of rats fed a combination of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion.
Detailed Description
The invention provides a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, which comprises porous starch and an artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound loaded in pores of the porous starch, wherein the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound comprises artemisinin and hydroxypropyl-beta-cyclodextrin coated on the surface of the artemisinin. In the invention, the hydroxypropyl-beta-cyclodextrin is a hydroxyalkyl derivative of the beta-cyclodextrin, has higher solubility and safety, has better tolerance to oral administration, intravenous injection, intradermal injection and parenteral administration, has high binding constant with an object, and increases the stability of a complex formed with the object. In the invention, the porous starch has good biocompatibility, easy degradation, low toxicity and strong adsorbability, and the porous starch is used as a carrier, thereby being beneficial to improving the bioavailability of the artemisinin.
In the invention, the content of artemisinin in the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound is preferably 10-30 wt%, more preferably 15-28 wt%, and further preferably 21.25-25.71 wt%, and the content of hydroxypropyl-beta-cyclodextrin is preferably 10-30 wt%, more preferably 16.86-28.48 wt%, and further preferably 23.31-24.58 wt%. In the present invention, the pore size of the porous starch is preferably less than 1 μm.
The invention provides a preparation method of a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, which comprises the following steps:
performing inclusion treatment on a first mixed system containing artemisinin and hydroxypropyl-beta-cyclodextrin to obtain an artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound;
and (3) adsorbing a second mixed system containing the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound and porous starch to obtain the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound.
In the present invention, unless otherwise specified, all the starting materials for the preparation are commercially available products well known to those skilled in the art.
The invention carries out inclusion treatment on a first mixed system containing artemisinin and hydroxypropyl-beta-cyclodextrin to obtain the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound. In the invention, the mass ratio of the hydroxypropyl-beta-cyclodextrin to the artemisinin is preferably (1.8-2.2): 1, more preferably 2: 1. in the present invention, the preparation method of the first mixed system preferably includes: and mixing the artemisinin solution with the hydroxypropyl-beta-cyclodextrin solution to obtain a first mixed system. In the invention, the mass content of the artemisinin in the artemisinin solution is preferably 4.5-85%, more preferably 20-60%, and further preferably 40-50%; the invention has no special limitation on the types of solvents in the artemisinin solution, and the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound obtained by subsequent inclusion treatment have good solubility and can be removed easily; in the present invention, the solvent in the artemisinin solution is preferably ethanol. In the invention, the mass content of the hydroxypropyl-beta-cyclodextrin in the hydroxypropyl-beta-cyclodextrin solution is preferably 4-85%, more preferably 20-60%, and further preferably 40-50%; the invention has no special limitation on the type of the solvent in the hydroxypropyl-beta-cyclodextrin solution, and the solvent which has good solubility and is easy to remove is only needed for the hydroxypropyl-beta-cyclodextrin and the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound obtained by subsequent inclusion treatment; in the present invention, the solvent in the hydroxypropyl-beta-cyclodextrin solution is preferably water, and more preferably deionized water. The feeding method for mixing the artemisinin solution and the hydroxypropyl-beta-cyclodextrin solution is not particularly limited, and specifically, the hydroxypropyl-beta-cyclodextrin solution is added into the artemisinin solution to obtain a first mixed system.
In the invention, the temperature of the inclusion treatment is preferably 40-60 ℃, and specifically can be 40 ℃, 45 ℃, 50 ℃, 55 ℃ or 65 ℃; the time is preferably 1-6 h, and specifically can be 1h, 2h, 3h, 4h, 5h or 6 h.
After the inclusion treatment, the solvent in the obtained system is preferably removed, and the remainder is washed and dried in sequence to obtain the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound. In the present invention, the solvent in the system obtained after the inclusion treatment is preferably removed by evaporation, and the present invention does not specifically limit the specific operating conditions for the evaporation, and any method known to those skilled in the art may be used. In the present invention, the reagent used for washing is preferably water, more preferably deionized water, and the present invention preferably washes until the deionized water is clear to ensure that the non-included hydroxypropyl-beta-cyclodextrin is sufficiently removed. The drying mode and conditions are not particularly limited, sufficient drying is guaranteed, the property and the appearance of the substance are not changed, and particularly, the drying mode is preferably drying or freeze drying.
After the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound is obtained, a second mixed system containing the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound and porous starch is subjected to adsorption treatment, and the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound is obtained. In the invention, the mass ratio of the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound to the porous starch is preferably 1: (0.8 to 1.2), more preferably 1: 1. in the present invention, the preparation method of the second mixed system preferably includes: and mixing the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound with the porous starch dispersion system to obtain a second mixed system. In the invention, the mass content of the porous starch in the porous starch dispersion system is preferably 30-70%, more preferably 33-67%, and further preferably 45-50%; the invention has no special limitation on the type of a dispersion medium in the porous starch dispersion system, and the invention only needs the reagent which does not dissolve the porous starch, has poor solubility to the artemisinin and is easy to remove; in the present invention, the dispersion medium in the porous starch dispersion is preferably water, more preferably deionized water. The feeding mode of the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound and the porous starch dispersion system is not particularly limited, and the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound can be added into the porous starch dispersion system.
In the invention, the temperature of the adsorption treatment is preferably 30-40 ℃, and specifically can be 30 ℃, 37 ℃ or 40 ℃; the time is preferably 0.5-2 h, and specifically can be 0.5h, 1h or 2 h.
After the adsorption treatment, the invention preferably performs solid-liquid separation on the obtained system, and dries the obtained solid material to obtain the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound. The solid-liquid separation mode is not particularly limited, and the mode of effectively separating target substances without damaging the structure of the target substances can be specifically filtration or centrifugation; the rotation speed of the centrifugation is preferably 7000-9000 r/min, and more preferably 8000 r/min; the time is preferably 8-12 min, and more preferably 10 min. The drying mode and conditions are not particularly limited, sufficient drying is guaranteed, the property and the appearance of the substance are not changed, and particularly, the drying mode is preferably drying or freeze drying.
The method provided by the invention has the advantages of simple process flow, short preparation period and easy industrial production; meanwhile, the solvents used in the preparation of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound are non-toxic or low-toxicity, do not easily harm experimenters and do not easily pollute the environment; the required equipment is simple, the price is low, and the production cost is low.
The porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound prepared by the method provided by the invention has the preferable drug loading rate of 10.12-20.47% and the preferable encapsulation rate of 13.43-25.11%. In the invention, the artemisinin is in a crystal morphological structure, the porous starch is in an amorphous state and is loaded on the porous starch after being included by the hydroxypropyl-beta-cyclodextrin, and the obtained porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound shows the same amorphous state as the porous starch, thereby improving the water solubility of the artemisinin.
The invention provides application of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in the technical scheme or the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound prepared by the preparation method in the technical scheme in preparation of a drug for treating malaria. In the present invention, the drug for treating malaria preferably comprises an active ingredient and pharmaceutically acceptable excipients; the invention is not limited to the type of the pharmaceutically acceptable excipients, and the excipients well known to those skilled in the art can be used. In the present invention, the dosage form of the drug for treating malaria preferably includes an oral preparation; the drug for treating malaria is particularly suitable for treating cerebral malaria, and particularly, the dosage form of the drug for treating malaria can be an intracerebral distribution oral preparation.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
(1) Mixing artemisinin with ethanol to obtain artemisinin solution with mass content of 50%; mixing hydroxypropyl-beta-cyclodextrin with deionized water to obtain a hydroxypropyl-beta-cyclodextrin solution with the mass content of 50%; according to the mass ratio of the hydroxypropyl-beta-cyclodextrin to the artemisinin being 2: 1, adding hydroxypropyl-beta-cyclodextrin solution into artemisinin solution, and performing inclusion treatment for 3 hours at 40 ℃; evaporating the obtained system to remove the solvent, washing the residue with deionized water, and freeze-drying to obtain artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound;
(2) mixing porous starch (the aperture is less than 1 mu m) and deionized water according to the mass ratio of 1: 1 to obtain a porous starch dispersion system; according to the mass ratio of the porous starch to the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound of 1: 1, adding the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound into a porous starch dispersion system, and performing adsorption treatment for 30min at the temperature of 30 ℃; centrifuging the obtained system (8000r/min, 10min), and freeze-drying the obtained solid material to obtain porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, which is recorded as AHPS; the drug loading rate of the AHPS is 10.12 percent, and the content of hydroxypropyl-beta-cyclodextrin is 23.31 percent; the encapsulation efficiency was 13.43%.
Example 2
(1) Mixing artemisinin with ethanol to obtain artemisinin solution with mass content of 50%; mixing hydroxypropyl-beta-cyclodextrin with deionized water to obtain a hydroxypropyl-beta-cyclodextrin solution with the mass content of 50%; according to the mass ratio of the hydroxypropyl-beta-cyclodextrin to the artemisinin being 2: 1, adding hydroxypropyl-beta-cyclodextrin solution into artemisinin solution, and performing inclusion treatment for 1h at the temperature of 60 ℃; evaporating the obtained system to remove the solvent, washing the residue with deionized water, and freeze-drying to obtain artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound;
(2) mixing porous starch (the aperture is less than 1 mu m) and deionized water according to the mass ratio of 2: 1 to obtain a porous starch dispersion system; according to the mass ratio of the porous starch to the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound of 1: 1, adding the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound into a porous starch dispersion system, and performing adsorption treatment for 60min at 37 ℃; centrifuging the obtained system (8000r/min, 10min), and freeze-drying the obtained solid material to obtain porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, which is recorded as AHPS; the drug loading rate of the AHPS is 12.02 percent, and the content of hydroxypropyl-beta-cyclodextrin is 16.86 percent; the encapsulation efficiency was 16.84%.
Example 3
(1) Mixing artemisinin with ethanol to obtain artemisinin solution with mass content of 50%; mixing hydroxypropyl-beta-cyclodextrin with deionized water to obtain a hydroxypropyl-beta-cyclodextrin solution with the mass content of 50%; according to the mass ratio of the hydroxypropyl-beta-cyclodextrin to the artemisinin being 2: 1, adding hydroxypropyl-beta-cyclodextrin solution into artemisinin solution, and performing inclusion treatment for 5 hours at the temperature of 45 ℃; evaporating the obtained system to remove the solvent, washing the residue with deionized water, and freeze-drying to obtain artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound;
(2) mixing porous starch (the aperture is less than 1 mu m) and deionized water according to the mass ratio of 1: 2 to obtain a porous starch dispersion system; according to the mass ratio of the porous starch to the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound of 1: 1, adding the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound into a porous starch dispersion system, and performing adsorption treatment for 120min at the temperature of 40 ℃; centrifuging the obtained system (8000r/min, 10min), and freeze-drying the obtained solid material to obtain porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, which is recorded as AHPS; the drug loading rate of the AHPS is 20.47 percent, and the content of hydroxypropyl-beta-cyclodextrin is 24.58 percent; the encapsulation efficiency was 25.11%.
Characterization and Performance testing
The porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound prepared in example 3 was characterized as follows:
FIG. 1 is an SEM picture of artemisinin (a), hydroxypropyl-beta-cyclodextrin (b), artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (c), porous starch (d) and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (e, f). As can be seen from a in fig. 1, due to its lipid solubility, artemisinin is recrystallized from a monomolecular state of a true solution to form crystals with a particle size of more than 1mm, which are irregular long columns and form a water-insoluble suspension system in water; as can be seen from b in FIG. 1, hydroxypropyl-. beta. -cyclodextrin is hollow cylindrical and suitable as an inclusion material; as can be seen from c in fig. 1, the surface of the porous starch has a porous interconnected structure, and the pore diameter is less than 1 μm; as can be seen from d in FIG. 1, the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound is an irregular complex formed after artemisinin is included by hydroxypropyl-beta-cyclodextrin. As can be seen from e and f in figure 1, after the porous starch adsorbs the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, the particle size of the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in the pores of the porous starch is less than 100nm, the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound can form a colloidal solution with water, and the water solubility is good.
FIG. 2 is an XRD pattern of artemisinin (A), hydroxypropyl-beta-cyclodextrin (B), porous starch (C) and porous starch loaded hydroxypropyl-beta-cyclodextrin inclusion compound (D). As can be seen from fig. 2, the diffraction peaks in the XRD pattern of the porous starch sample show extremely weak peaks at two positions, 2 θ 18.32 ° and 2 θ 22.12 °, respectively, indicating that the porous starch is in an amorphous structure and exists in an amorphous form. Diffraction peaks in the XRD pattern of the artemisinin sample are at 2 theta of 7.32 degrees, 11.73 degrees, 14.78 degrees, 22.24 degrees, 38.89 degrees and 42.06 degrees respectively, and obvious diffraction peaks exist at six positions, which indicates that the artemisinin has a crystal structure and exists in a crystal form. Diffraction peaks in XRD patterns of the sample of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound are respectively at positions of 2 theta (7.5 degrees), 12.5 degrees, 15.1 degrees, 21.1 degrees and 22.5 degrees, obvious diffraction peaks are formed at the five positions, and the XRD pattern of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound shows weaker diffraction peaks compared with artemisinin. The hydroxypropyl-beta-cyclodextrin and the porous starch have three wider peaks within the ranges of 2 theta (13-16 degrees), 16-20 degrees and 21-25 degrees, which shows that the hydroxypropyl-beta-cyclodextrin and the porous starch are basically in an amorphous state. The X-ray diffraction pattern of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound is basically consistent with that of the porous starch, and weak characteristic peaks of phloretin are shown only at positions of 7.5 degrees and 15.1 degrees of 2 theta, and the result proves again that artemisinin is completely prepared into the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, so that the amorphous structure similar to that of the porous starch is shown, and the water solubility of artemisinin is improved.
The performance test of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound prepared in example 3 is as follows:
1. and (3) testing the dissolvability in water, which comprises the following specific steps:
accurately measuring 200mL of deionized water as a dissolution medium, and putting the deionized water into a water bath kettle with a stirrer, wherein the water bath temperature is as follows: 37.0 +/-0.5 ℃, and the stirring speed is as follows: 100 r/min; respectively adding the weighed 62.5mg artemisinin original drug, 62.5mg dihydroartemisinin, 300.33mg porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin clathrate and 520mg artemisinin piperaquine tablet (containing artemisinin 62.5mg) into dissolution medium, and respectively sampling 1mL at the time points of 0.083h, 0.17h, 0.25h, 0.33h, 0.42h, 0.5h, 0.58h, 0.67h, 0.75h, 0.83h, 0.92h, 1h, 1.17h, 1.33h, 1.5h, 1.67h, 1.83h, 2h, 2.5h, 3h, 3.5h, 4h, 4.5h, 5h, 6h, 7h, 8h, 9h, 10h, 11h, 12h, 24h, 28h, 32h, 36h and 48h (1 mL of dissolution medium is added again after 1mL is taken out every time), centrifuging the taken out samples respectively (10000r/min, 5min), taking out 10 mu L of supernatant respectively, and detecting the content of artemisinin.
Fig. 3 is a dissolution curve diagram of porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (A), dihydroartemisinin (B), artemisinin piperaquine tablet (C) and artemisinin original drug (D) in water. As can be seen from figure 3, the release effect of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in water is obviously improved compared with that of artemisinin original drug, dihydroartemisinin and artemisinin piperaquine tablets. The final release cumulant of the artemisinin original drug, the dihydroartemisinin and the artemisinin piperaquine tablet in water is 19.75%, 32.24% and 24.21%, respectively, while the final release cumulant of the porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in water is 61.25%, which is about 3.10 times, about 1.90 times and about 2.53 times of the final release cumulant of the artemisinin original drug.
2. The method for testing the dissolubility in the artificial gastric juice comprises the following specific steps:
the dissolution test in water is carried out according to the method, and the difference is only that the dissolution medium is artificial gastric juice simulating gastric juice environment.
Fig. 4 is a dissolution curve diagram of porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (A), dihydroartemisinin (B), artemisinin piperaquine tablet (C) and artemisinin original drug (D) in artificial gastric juice. As can be seen from figure 4, the release effect of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in artificial gastric juice is obviously improved compared with that of artemisinin original drug, dihydroartemisinin and artemisinin piperaquine tablet. The final release cumulant of the artemisinin original drug, the dihydroartemisinin and the artemisinin piperaquine tablet in the artificial gastric juice is 24.27%, 31.94% and 25.79% respectively, and the final release cumulant of the porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in the artificial gastric juice is 61.64%, about 2.54 times, about 1.93 times and about 2.39 times of the final release cumulant of the artemisinin original drug.
3. The method for testing the dissolubility of the artificial intestinal juice comprises the following specific steps:
the method is carried out according to the method of 'dissolution in water' test, and the difference is only that the dissolution medium is artificial intestinal juice simulating an intestinal juice environment.
Fig. 5 is a dissolution curve diagram of porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound (A), dihydroartemisinin (B), artemisinin piperaquine tablet (C) and artemisinin original drug (D) in artificial intestinal juice. As can be seen from figure 5, the release effect of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in the artificial intestinal juice is obviously improved compared with the release effect of artemisinin original drug, dihydroartemisinin and artemisinin piperaquine tablet. The final release accumulation amounts of the artemisinin original drug, the dihydroartemisinin and the artemisinin piperaquine tablet in the artificial intestinal juice are only 1.650 percent, 1.810 percent and 1.880 percent respectively, and the final release accumulation amount of the porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in the artificial intestinal juice is 2.223 percent, about 1.35 times, about 1.23 times and about 1.18 times of the final release accumulation amount of the artemisinin original drug.
4. The test of the protective effect of the CM rat comprises the following specific steps:
establishing a plasmodium boehmeri strain mouse malaria model: the infected C57BL/6 rat is inoculated with Pb ANKA strain in abdominal cavity, and when the protozoa blood level of the rat reaches about 30%, 1 × 10 rats are taken7(iii) intraperitoneal inoculation of pRBC into offspring C57BL/6 rats, scored as day 0 of inoculation; randomly dividing C57BL/6 rats into 5 groups, namely a Model group (Model), an artemisinin original drug group (ART), a porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound group (AHPS), a dihydroartemisinin group (DHA) and an artemisinin piperaquine group, wherein each group comprises 10 artemisinin groups; the dihydroartemisinin group was inoculated at 7.8 mg-kg from day 0-1Intragastric administration, arteannuin protodrug group and porous starch loaded arteannuin hydroxypropylThe content of the (E) -beta-cyclodextrin inclusion compound is 7.8 mg/kg according to the artemisinin content-1The tablet is administered by intragastric administration, and the content of artemisinin in the tablet is 139.10 mg/kg-1Intragastric administration, 1 time per day, and continuous administration for 4 days; protozoa were calculated every day starting from 3d infection of experimental rats; taking 1 mu L of infected rat tail venous blood to prepare a thin blood sheet on a clean glass slide, fixing and air-drying by using methanol, staining by using Giemsa staining solution for 20min, washing and air-drying, counting 1000 erythrocytes under an X100 oil scope, and counting protozoan blood as pRBC/total erythrocyte number; when the drug is administered for 4 days, the morphological change of the plasmodium is microscopically examined under an X100 oil microscope, and the pictures are shot by a digital camera; the survival condition of the rats is observed day by day, after 14 days, a survival curve and the level change of protozoan blood are drawn, and the survival rate is equal to that of survival experimental rats/total experimental rats multiplied by 100%.
FIG. 6 is a graph showing the results of the porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound, dihydroartemisinin, artemisinin piperaquine tablets and artemisinin original drug on the infection rate of CM rats. As can be seen from FIG. 6, the protozoan blood level was significantly reduced (P < 0.001) in the ART group, DHA group and AHPS group at day 4 of infection, and significantly lower in the AHPS group than in the Art group and DHA group at day 9-10 of infection. The model group started to develop coma and death by day 6, with a mortality rate of 50% by day 14 of vaccination. In contrast, only 2 of the ART groups died from CM on day 10 of vaccination with a mortality of 20% by day 14 of vaccination. The DHA group had 3 deaths due to CM starting from day 9 of inoculation and 30% mortality by day 14 of inoculation. All AHPS groups survived with 0% mortality by day 14 of vaccination. Due to the long half-life of piperaquine, acute toxicity occurred after 4 consecutive days of administration, 2 deaths occurred on the 5 th day of inoculation and recovered after drug withdrawal, with a mortality rate of 20% by the 14 th day of inoculation. It can be seen that AHPS has significant protective effect on CM rats and has high safety.
5. The bioavailability test comprises the following specific steps:
140 SD rats (from animal center of first subsidiary hospital of Harbin medical university) with weight of 200-250 g were fed with standard pellet feed and freely drunk water; the temperature of the breeding environment is 18-23 ℃, and the humidity is kept at 45-70%. 135 female SD rats weighing 200-250 g are selected, randomly divided into five groups, each group comprises 27 animals, and treated by gastric lavage, and are separately infused with artemisinin original drug, dihydroartemisinin, artemisinin piperaquine tablet, porous starch loaded artemisinin (prepared by the method of step (2) in example 1, except that the 'artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound' is replaced by 'artemisinin') and porous starch loaded with artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound. Fasting was started 12h before gavage to ensure adequate drinking, and the gavage concentration of the four groups of rats was 50mg/kg (measured as artemisinin concentration). Counting from the time of gastric perfusion, taking blood from orbital veins of rats at 0.5h, 1.0h, 2.0h, 4.0h, 6.0h, 8.0h, 10.0h, 12.0h and 24.0h respectively, taking about 500 mu L of blood, putting into a centrifuge tube filled with 20 mu L of 1% heparin sodium, shaking gently to mix the heparin sodium with blood samples, centrifuging for 10min at 3000r/min, taking supernatant plasma, storing in a refrigerator at 4 ℃ and treating the same day. Precisely sucking 100 mu L of supernatant plasma into a 1.5mL centrifuge tube filled with 1% sodium heparin, adding 300 mu L of acetonitrile, vortexing for 3min, centrifuging (12000r/min, 15min), taking supernatant, drying with nitrogen, re-dissolving residues with 100 mu L of mobile phase (acetonitrile: water: 60: 40 in terms of volume ratio), vortexing for 1min, centrifuging (12000r/min, 15min), taking supernatant, filtering with a 0.22 mu m filter membrane, sucking 10 mu L of supernatant, and detecting by a high performance liquid chromatograph to obtain the artemisinin concentration in the sample.
Figure 7 is a graph of bioavailability results of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound fed to rats. As can be seen from fig. 7, when rats were gavaged under the condition of the same content of artemisinin and the blood concentration of rats was measured 24h after gavage, the artemisinin protogroup, the dihydroartemisinin group, the porous starch-loaded artemisinin group and the artemisinin piperaquine group could not detect the retention of the drugs in vivo basically, and the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound could still detect the blood concentration 24h after gavage. The result shows that the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound has longer metabolism time in a rat body, so that the drug effect is stronger, the sustained release effect is achieved, and the bioavailability of artemisinin is greatly improved by the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound.
The artemisinin concentration in each tissue of the rat is tested, and the specific steps are as follows:
rats began to fast 12h before gavage to ensure adequate drinking. Before gavage, rats of each group were weighed and body weights recorded. Each group of 13 rats was randomly divided into six groups, of which 3 were intragastric artemisinin original drug groups, 3 were intragastric porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion group, 3 were intragastric artemisinin piperaquine group, 3 were intragastric dihydroartemisinin group, and 1 was blank control group. The amount of the gavage is 250mg/kg (calculated according to the content of the artemisinin), the rats are respectively and rapidly killed and dissected after 0.5h, 1h, 2h, 4h, 6h, 8h, 12h and 24h of gavage, the total 6 organs of the heart, the liver, the spleen, the lung, the kidney and the brain are picked up, the picked-up organs are respectively placed in normal saline with marked serial numbers, residual blood in the organs is repeatedly cleaned until the organs do not show blood red when the filter paper absorbs water, the normal saline on the organs is completely sucked and dried by the filter paper, the organs are placed in a sealing belt with marked serial numbers, and liquid nitrogen is uniformly frozen and stored in a refrigerator at minus 80 ℃ for freezing storage.
The following treatments were performed on the rat organs taken out in this experiment: after 6 organ tissues in total, i.e., heart, liver, spleen, lung, kidney and brain, of rats were precisely weighed using a BSA124S-CW type electronic analytical balance, the organs were placed in test tubes, and homogenized at a high speed after adding 1mL of physiological saline, and the concentration of the resulting tissue homogenate was 200 mg/mL. Adding the accurately sucked 100 mu L of tissue homogenate into a centrifuge tube, adding 1mL of acetonitrile, sealing a sealing film, uniformly mixing by a vortex mixer, performing vortex for 1min, performing ultrasonic treatment for 10min, and centrifuging for 10min at 13000 r/min. And taking the supernatant, and uniformly storing in a refrigerator at 4 ℃ for cold storage. After the amount of the samples is certain, detecting and analyzing the content of the artemisinin and the dihydroartemisinin in each tissue and organ by using a high performance liquid chromatograph.
FIGS. 8-15 are graphs showing the comparison results of the concentration of artemisinin in tissues after feeding rats with artemisinin original drug, dihydroartemisinin, artemisinin piperaquine tablet and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound for 0.5h, lh, 2h, 4h, 6h, 8h, 12h and 24 h. As shown in fig. 8 to fig. 15, the artemisinin original drug, dihydroartemisinin, porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound and artemisinin piperaquine tablet have distribution in heart, liver, spleen, lung, kidney and brain tissues of rats after being perfused for 30min, and for each time point, the concentration of artemisinin is higher in liver and heart, then kidney and lung, and has a small distribution in brain, and the concentration is lowest in spleen. And the average content of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in each tissue at different time points is higher than that of artemisinin original drug, dihydroartemisinin and artemisinin piperaquine tablets.
Figure 16 is a graph of bioavailability results in the heart of rats fed a combination of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion. As can be seen from FIG. 16, the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion group has higher blood concentration and longer residence time in rat heart. The artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound loaded by the porous starch has smaller molecular particle size, and improves the dissolution rate and the slow release of the artemisinin, so that the absorption and utilization effects of the porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound after gastric lavage of rats are obviously better than those of artemisinin original drugs, dihydroartemisinin, porous starch loaded artemisinin and artemisinin piperaquine tablets.
Figure 17 is a graph of the bioavailability results in the liver of rats fed a combination of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion. As can be seen from FIG. 17, the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion group improves the water solubility of artemisinin in rat liver, and is more favorable for absorption and utilization. The artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound loaded by the porous starch has smaller molecular particle size, and improves the dissolution rate and the slow release of the artemisinin, so that the absorption and utilization effects of the porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound after gastric lavage of rats are obviously better than those of artemisinin original drugs, dihydroartemisinin, porous starch loaded artemisinin and artemisinin piperaquine tablets.
Figure 18 is a graph of bioavailability results in the spleen of rats fed a combination of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion. As can be seen from fig. 18, the artemisinin hydroxypropyl- β -cyclodextrin inclusion compound loaded with porous starch in the present invention has smaller molecular particle size, and improves the dissolution rate and sustained release of artemisinin, so that after gastric lavage of rats, the absorption and utilization effects of the porous starch loaded artemisinin hydroxypropyl- β -cyclodextrin inclusion compound are significantly better than those of artemisinin original drug, dihydroartemisinin, porous starch loaded artemisinin and artemisinin piperaquine tablets, and the spleen concentration is lower compared with the bioavailability parameter values of other organs, which is due to less spleen blood flow.
Figure 19 is a graph of bioavailability results in the lungs of rats fed a combination of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl- β -cyclodextrin inclusion. Since the alveolar epithelium is a lipid membrane, the absorption of artemisinin into the lungs is a passive diffusion process and deposits or losses occur everywhere in the respiratory tract, and thus the percentage of bioactive molecules reaching the alveoli is not high enough. But the bioavailability of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in rat lung is improved to a certain extent.
Figure 20 is a graph of bioavailability results in the kidney of rats fed a combination of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion. The porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound has smaller molecular particle size, and improves the solubility and slow release of artemisinin in water, so that the absorption and utilization effects of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound are obviously better than those of other medicines after the rats are subjected to intragastric administration; it is also known from the comparison that the concentration of artemisinin in the kidney is only after the heart and liver, the kidney is the main excretory organ for bioactive molecules, and the content of porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound in the kidney is obviously increased. Therefore, artemisinin can be better developed and utilized as an effective antimalarial bioactive molecule.
Figure 21 is a graph of bioavailability results in the brain of rats fed a combination of artemisinin bulk drug, dihydroartemisinin, artemisinin piperaquine tablets, porous starch loaded artemisinin and porous starch loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion. The blood brain barrier of the brain is due to the fact that endothelial cells in the brain capillaries only allow fat-soluble small molecules to pass through, while artemisinin is a small molecule that is poorly soluble in water, the lipid solubility of the arteannuin can enable the arteannuin to smoothly penetrate through a blood brain barrier to reach brain tissues, and the arteannuin is included by adopting hydroxypropyl-beta-cyclodextrin and then loaded by taking porous starch as a carrier, so that the particle size of arteannuin molecules and water can be reduced to form a colloid system, the water solubility of the artemisinin is improved on the basis of not changing the lipid solubility of the artemisinin, so that the amount of the artemisinin passing through a blood brain barrier is far more than that of an artemisinin original drug, dihydroartemisinin, porous starch loaded artemisinin and an artemisinin piperaquine tablet, therefore, after the rat is subjected to intragastric administration, higher concentrations of artemisinin were detected in their brains, which enabled the porous starch loaded hydroxypropyl- β -cyclodextrin inclusion complex of artemisinin to be more effective in the treatment of cerebral malaria.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound comprises porous starch and artemisinin hydroxypropyl-beta-cyclodextrin inclusion compounds loaded in pores of the porous starch, wherein the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound comprises artemisinin and hydroxypropyl-beta-cyclodextrin coated on the surface of the artemisinin.
2. The porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound of claim 1, wherein the content of artemisinin in the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound is 10-30%, and the content of hydroxypropyl-beta-cyclodextrin is 10-30%.
3. A process for the preparation of a porous starch loaded artemisinin hydroxypropyl- β -cyclodextrin inclusion complex as claimed in claim 1 or 2, comprising the steps of:
performing inclusion treatment on a first mixed system containing artemisinin and hydroxypropyl-beta-cyclodextrin to obtain an artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound;
and (3) adsorbing a second mixed system containing the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound and porous starch to obtain the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound.
4. The preparation method according to claim 3, wherein the mass ratio of the hydroxypropyl-beta-cyclodextrin to the artemisinin is (1.8-2.2): 1.
5. the method of manufacturing according to claim 3, wherein the method of manufacturing the first mixed system comprises: mixing the artemisinin solution with the hydroxypropyl-beta-cyclodextrin solution to obtain a first mixed system; the mass content of the artemisinin in the artemisinin solution is 4.5-85%, and the mass content of the hydroxypropyl-beta-cyclodextrin in the hydroxypropyl-beta-cyclodextrin solution is 4-85%.
6. The method according to any one of claims 3 to 5, wherein the temperature of the inclusion treatment is 40 to 60 ℃ and the time is 1 to 6 hours.
7. The preparation method according to claim 3, wherein the mass ratio of the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound to the porous starch is 1: (0.8 to 1.2).
8. The method of manufacturing according to claim 3, wherein the method of manufacturing the second mixed system comprises: mixing the artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound with the porous starch dispersion system to obtain a second mixed system; the mass content of the porous starch in the porous starch dispersion system is 30-70%.
9. The method according to claim 3, 7 or 8, wherein the temperature of the adsorption treatment is 30 to 40 ℃ and the time is 0.5 to 2 hours.
10. Use of the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound as claimed in claim 1 or 2 or the porous starch-loaded artemisinin hydroxypropyl-beta-cyclodextrin inclusion compound prepared by the preparation method as claimed in any one of claims 3 to 9 in preparation of a medicament for treating malaria.
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| CN114470230A (en) * | 2021-12-31 | 2022-05-13 | 中国科学院重庆绿色智能技术研究院 | Artemisinin water-soluble compound and preparation method and application thereof |
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