CN104622801A - Hydroxycamptothecine cubic liquid crystal precursor composition as well as preparation method and application thereof - Google Patents

Abstract

The invention discloses a cubic liquid crystal precursor composition of hydroxycamptothecin, a preparation method and application thereof, and belongs to the technical field of pharmacy. The composition is mainly prepared from the following raw materials in weight percentage: 4-6% hydroxycamptothecin, 50-78% liquid crystal material, 2-8% PEG, 1-3% oleic acid glyceride, 2 - 7% glyceryl stearate, 13-26% dispersion medium, wherein: the dispersion medium is composed of the following raw materials in weight percentage: 8-30% water-miscible organic solvent, 70-92% water. The composition can be continuously released in vitro for more than 30 days, greatly prolonging the action time of the hydroxycamptothecin, and due to the particularity of the local administration method, can simultaneously achieve the effects of sustained release and targeting.

Description

Cubic liquid crystal precursor composition of hydroxycamptothecin and its preparation method and application

technical field

The invention relates to the technical field of pharmacy, in particular to a cubic liquid crystal precursor composition of hydroxycamptothecin and its preparation method and application.

Background technique

Hydroxycamptothecin is a tryptophan-terpene alkaloid isolated from Campylodontia involucratae, which inhibits DNA replication, transcription and mitosis by inhibiting DNA topoisomerase Ⅰ, thereby making tumor cells Lethal damage occurs, and it is mainly used clinically for malignant tumors such as gastric cancer, primary liver cancer, leukemia, rectal cancer, and bladder cancer.

The antitumor activity of hydroxycamptothecin is closely related to its molecular structure. Closing the lactone ring is a necessary form to exert anticancer activity, and opening the lactone ring will greatly reduce or inactivate the activity. Studies have shown that the existing form of the camptothecin drug structure depends on the acidity and alkalinity of the system. In the blood plasma at pH 7.4, the lactone ring is rapidly hydrolyzed and opened into an inactive carboxyl form, and the half-life is only about 5 minutes. Only 0.2% are in the form of ester rings.

At present, hydroxycamptothecin is mainly used clinically in the form of sodium salt injection, sodium salt powder injection and capsules. The phenolic hydroxyl group of sodium salt injection is unstable in the air or exposed to light or heat, and is easily oxidized and hydrolyzed, and the content decreases. At the same time, the opening of the lactone ring reduces the curative effect. However, there is still the problem of reduced curative effect of lactone ring-opening and short half-life. Moreover, after intravenous administration of sodium salt injection and sodium salt powder injection, they are mainly distributed in the contents of the gallbladder and small intestine, and cancer often occurs in tissues and organs. For example, the distribution in tissues such as liver and lung is not ideal; in addition, although capsules can be administered orally, they are suitable for treating gastrointestinal tumors, but the increased dosage increases the toxic and side effects on the gastrointestinal tract. In summary, factors such as fast metabolism of hydroxycamptothecin, unsatisfactory tissue distribution and instability of the lactone ring hinder its full anti-tumor effect and limit its clinical application.

In order to overcome the above defects of hydroxycamptothecin preparations, researchers at home and abroad have used modern pharmaceutical methods to prepare various hydroxycamptothecin drug-loading systems to improve their solubility and lactone ring stability, prolong blood circulation time and reduce toxicity. Side effects have been reported in the form of microspheres, emulsions, micelles, liposomes, nanoparticles and vesicles.

Some studies have made hydroxycamptothecin into liposomes or nanoparticles to prolong its circulation time in the body. However, due to the body's defense function, hydroxycamptothecin accumulates in the reticuloendothelial system in large quantities, and it is easy to damage the non-pathological reticulum. The endothelial system produces toxicity, and liposomes are colloidal drug delivery systems, which are prone to instability phenomena such as flocculation, aggregation, delamination, particle size increase, and drug leakage during placement; and the stability of nanoparticles in the solution phase Not high, hydroxycamptothecin is easy to precipitate, although the method of freeze-drying can improve the stability, but it is easy to reduce the encapsulation efficiency, the problem that the particle size of the nanoparticle becomes larger, and the preparation process of the nanoparticle is mostly carried out under alkaline conditions, It has a great influence on the stability of hydroxycamptothecin.

In recent years, hydroxycamptothecin microemulsion is also a research hotspot, but the targeting of microemulsion is poor. The microemulsion system has certain irritation and safety, and whether it can maintain the original stability when diluted is an urgent investigation. The problem. In addition, although the double membrane structure of the vesicle can well contain hydroxycamptothecin and has good compatibility with the cell membrane, the fusion between the membranes of the vesicle is prone to occur, and its stability needs to be improved; hydroxycamptothecin Alkaline micelles how to avoid lysosomes and break through the barriers of the nuclear membrane after reaching the cells have yet to be resolved; although the carrier of solid lipid nanoparticles can reduce systemic toxicity, it is non-selective in important organs and lacks corresponding toxicity. Data is the problem with this drug delivery system.

At present, the existing technology has not yet solved the pharmaceutical problems of poor stability and poor water solubility of hydroxycamptothecin, and the biopharmaceutical problem of short half-life of hydroxycamptothecin. In order to make the drug more stable and long-term in the blood In order to further improve drug efficacy and reduce adverse reactions, it is still necessary to study the drug delivery system to solve the limitations of the current hydroxycamptothecin carriers, and to find a more effective drug delivery system to supplement the drug delivery system. Self-deficiency, in order to solve the problem of its low clinical curative effect.

Current studies have found that a certain concentration of amphiphilic lipids will spontaneously form a thermodynamically stable lipid bilayer in water, and then recombine into cubic liquid crystals with various shapes and structures, which is the closure of bicontinuous water domains and lipid domains. Lipid bilayer "honeycomb (sponge)" structure, can maintain stability at higher temperature, and its internal surface area, viscosity and membrane strength are large, these characteristics make cubic liquid crystal become a good drug carrier. According to reports, as a drug carrier, cubic liquid crystals have many advantages: drug encapsulation diversity, it can encapsulate both water-soluble drugs and fat-soluble drugs, protein drugs hemoglobin and insulin can also be encapsulated in it; Biocompatibility and adhesion; improve drug stability, such as preventing protein and polypeptide drugs from being enzymatically hydrolyzed, protecting such drugs from external interference, so as to maintain their biological activity; good slow and controlled release effect.

There are three main forms of cubic liquid crystal as a drug carrier: precursor, gel and particle dispersion system. The precursor exists in solid or liquid form, and forms cubic liquid crystals under the induction of external factors such as contact with liquids; gel is an optically isotropic liquid crystal, viscous and transparent, similar in appearance to solids, and can coexist in equilibrium with water. Skin drug delivery system; liquid crystal particle dispersion system is that the liquid crystal material and water form cubic liquid crystals and then disperse in the excess aqueous solution in nanometer size.

Contents of the invention

Based on this, the object of the present invention is to provide a cubic liquid crystal precursor composition of hydroxycamptothecin for the defects of hydroxycamptothecin resin insoluble, water-insoluble, and easy to open and deactivate with alkali, and hydroxycamptothecin Encapsulated in cubic liquid crystals and made into a precursor composition, the composition has a long release time and stable release, and the active ingredient is not easy to be deactivated by ring opening.

To achieve the above object, the present invention takes the following technical solutions:

A cubic liquid crystal precursor composition of hydroxycamptothecin, which is mainly prepared from the following raw materials in weight percentage:

Wherein: dispersion medium is made up of the raw material of following percentage by weight:

Organic solvent miscible with water 8-30%

Water 70-92%.

In the research of the in-situ type cubic liquid crystal drug-loaded system, the inventors found that during the release process of the in-situ type cubic liquid crystal drug-loaded system, the composition ratio of the prescription changed due to the absorption of water and the release of the drug. This may cause a transition of the liquid crystal phase. Different liquid crystal phases have different kinetic mechanisms and release rates of drug release. For example, compared with cubic liquid crystals, the spatial structure of reversed-phase hexagonal liquid crystals will accelerate the release of insoluble drugs, but double delay the release of water-soluble drugs. Drug release, and the two-dimensional structure of lamellar liquid crystals will accelerate drug release. Therefore, if there is a liquid crystal phase transition during the release process, it will greatly affect the release rate and the stability of the release behavior.

After a large amount of experimental studies, the present inventor added glyceryl oleate and glyceryl stearate to the cubic liquid crystal precursor composition of the present invention, wherein glyceryl oleate is an amphiphilic lipid structure and has relatively Long hydrophobic hydrocarbon chains, on the one hand, do not affect the integrity of the cubic liquid crystal drug delivery system, on the other hand, the hydrophobic binding force of the hydrocarbon chains can strengthen the hydrophobic regions in the cubic liquid crystals, and improve the overall hydrophobicity of the precursor composition , to stabilize the critical packing parameters of the precursor composition, ensure that the system releases the drug slowly with the cubic liquid crystal carrier during the drug release process, and maintain the stability of the cubic liquid crystal; glyceryl stearate is hydrophobic and synergistic with glyceryl oleate The role is to facilitate the formation of cubic liquid crystals, which can more effectively control the drug release and make it difficult to undergo phase transition during the release process. The above-mentioned glyceryl oleate and glyceryl stearate are combined with hydrophilic PEG, and the three Work together to ensure the stability of the release rate and release behavior.

In one embodiment, the glyceryl oleate is glyceryl dioleate. Using glyceryl dioleate can make the obtained cubic liquid crystal precursor composition have better stability.

In one embodiment, the PEG is PEG400. In the investigation of many cubic liquid crystal systems of hydroxycamptothecin, it is found that because hydroxycamptothecin is a kind of insoluble drug, its release rate is difficult to reach 100%, and generally the maximum can be close to 70-80%, which is not conducive to bioavailability In order to improve the degree of drug release, hydrophilic substances can also be added to reduce the distribution coefficient of hydroxycamptothecin in cubic liquid crystals.

However, the addition of hydrophilic substances may lead to changes in the in-situ properties of the cubic liquid crystal precursor composition. The above-mentioned PEG400 can be used to reduce the in-situ performance of the cubic liquid crystal precursor composition. The distribution coefficient in cubic liquid crystals, and the hydrophilic nature of PEG400 can also promote the diffusion behavior of drugs, thereby enhancing the release of drugs and improving the release characteristics of drugs.

In one embodiment, the glyceryl stearate is selected from at least one of polyethylene glycol glyceryl stearate with an HLB value of 1 and a melting point ranging from 33°C to 43°C, and the stearin The weight percent of fatty acid glyceride is 2-4%. The melting point range of the HLB value of 1 is 33 DEG C-43 DEG C macrogol stearate such as Gelucire 33/01, Gelucire39/01, Gelucire 43/01 produced by Gattefosse Corporation wait.

In the research of the in-situ type cubic liquid crystal drug-loading system, the inventors found that the in-situ type cubic liquid crystal drug-loading system has a typical burst release phenomenon, that is, part of the drug is released rapidly during the in-situ process, thereby affecting the blood drug. Concentrations plateau and may exceed the therapeutic window. Select glyceryl stearate of the above-mentioned type for use, its HLB value is 1, hydrophobicity is strong, and melting point is lower, has the effect of enhancing the hydrophobicity of the precursor composition and being easy to mix evenly with other components in the precursor composition at body temperature The advantage is to achieve the effect of inhibiting the burst release of hydroxycamptothecin and stabilizing the crystal form, reducing the burst release amount in 24 hours from 40% to about 10%, and avoiding the release instability caused by burst release and crystal form change.

In one embodiment, the water-miscible organic solvent is selected from at least one of propylene glycol, N-methylpyrrolidone, 2-pyrrolidone and absolute ethanol.

In one embodiment, the liquid crystal material is phytantriol and/or glycerol monooleate. The two have less water absorption to form cubic liquid crystals, so as to achieve the effect of shortening the liquid crystal formation time. And the binary phase diagram shows that it has the advantages of being able to form and stabilize cubic liquid crystals at body temperature after being fully swollen, and the formed cubic liquid crystals can coexist with excess liquid.

The invention also discloses a preparation method of the above-mentioned cubic liquid crystal precursor composition of hydroxycamptothecin, comprising the following steps: dissolving hydroxycamptothecin in the dispersion medium according to the prescription amount, adding liquid crystal materials and mixing evenly, and then separately Add PEG, glyceryl oleate and glyceryl stearate and mix uniformly to obtain a cubic liquid crystal precursor composition of hydroxycamptothecin.

The invention also discloses the application of the above-mentioned cubic liquid crystal precursor composition of hydroxycamptothecin in the preparation of cubic liquid crystal preparation of hydroxycamptothecin.

In one of the embodiments, the hydroxycamptothecin cubic liquid crystal preparation is an injection.

In one of the embodiments, the injection is administered by intratumoral and/or paratumoral injection. Targeted drug administration is achieved, and the purpose of reducing systemic toxic and side effects is achieved.

Compared with the prior art, the present invention has the following beneficial effects:

A cubic liquid crystal precursor composition of hydroxycamptothecin of the present invention includes hydroxycamptothecin in cubic liquid crystals and makes a precursor composition, and hydroxycamptothecin is obtained by screening different liquid crystal materials and auxiliary materials. It is entrapped in the cubic liquid crystal precursor composition in the form of lactone, and does not precipitate after being placed, which ensures the in vitro stability of the drug and avoids the reduction of drug efficacy; in addition, due to the unique honeycomb structure of the cubic liquid crystal, it can maintain the hydroxycamptothecin The alkali is stable in the body and can be continuously released in vitro for more than 30 days, which greatly prolongs the action time of hydroxycamptothecin, and due to the particularity of local administration, it can achieve the effect of sustained release and targeting at the same time; in addition, The in vitro cytotoxicity experiment proves that the toxicity of the hydroxycamptothecin cubic liquid crystal precursor composition to cells is far greater than that of the hydroxycamptothecin injection, which obviously improves the curative effect of the hydroxycamptothecin.

Moreover, since the cubic liquid crystal structure is a good reservoir for hydroxycamptothecin, its drug loading capacity is ideal and can reach 6%. On the other hand, the addition of PEG400 and Gelucire also improves the overall drug resistance of the cubic liquid crystal affinity, so as to achieve the effect of solubilizing the drug.

The preparation method of the cubic liquid crystal precursor composition of hydroxycamptothecin of the present invention has the advantages of simple process, easy operation, strong controllability, good reproducibility, low cost, etc., and is easy for industrialized production.

The cubic liquid crystal precursor composition of hydroxycamptothecin of the present invention is prepared into a preparation, which effectively protects the active group of the drug, increases the stability of the drug, and improves the drug effect. In particular, it is made into an injection, which can be directed to the tumor target and realize the targeted drug delivery of hydroxycamptothecin.

Description of drawings

Fig. 1 is the schematic diagram of cytotoxicity of hydroxycamptothecin cubic liquid crystal precursor composition and hydroxycamptothecin injection in Experimental Example 1;

Fig. 2 is the drug release curve of part of the hydroxycamptothecin cubic liquid crystal precursor composition in Experimental Example 2;

Fig. 3 is the chromatogram that hydroxycamptothecin has ring-opening and ring-closing forms in Experimental Example 3;

Fig. 4 is the chromatogram that hydroxycamptothecin only has closed ring form in experimental example 3;

Fig. 5 is the SAXS spectrogram of the hydroxycamptothecin cubic liquid crystal precursor composition of Example 1-7 in Experimental Example 4;

Fig. 6 is the SAXS spectrum of the hydroxycamptothecin cubic liquid crystal precursor composition of Examples 8-9 and Comparative Examples 1-2 in Experimental Example 4.

Detailed ways

The present invention is described in detail below in conjunction with the drawings and specific embodiments, but does not impose any limitation on the present invention.

In the following examples:

Gelucire 33/01, Gelucire 39/01, Gelucire 43/01 were purchased from: Gattefosse Corporation.

Liver cancer cell Hep G2 cells, source: Institute of Clinical Pharmacology, School of Pharmacy, Sun Yat-sen University.

Hydroxycamptothecin injection, source: purchased from Hubei Huangshi Pharmaceutical Co., Ltd.

Example 1

A cubic liquid crystal precursor composition of hydroxycamptothecin, prepared from the following raw materials:

The cubic liquid crystal precursor composition of above-mentioned hydroxycamptothecin is prepared by the following method:

Weigh hydroxycamptothecin and the dispersion medium absolute ethanol according to the above formula, disperse and dissolve hydroxycamptothecin in absolute ethanol, slowly add water dropwise to the above solution, then add phytantriol, vortex and mix evenly, Then PEG 400, glyceryl dioleate and Gelucire 33/01 were added to the mixture, and the hydroxycamptothecin cubic liquid crystal precursor composition A was obtained after vortex mixing at room temperature.

Example 2

A cubic liquid crystal precursor composition of hydroxycamptothecin, prepared from the following raw materials:

The cubic liquid crystal precursor composition of above-mentioned hydroxycamptothecin is prepared by the following method:

Weigh hydroxycamptothecin and propylene glycol as the dispersion medium according to the above formula, disperse and dissolve hydroxycamptothecin in propylene glycol, slowly add water dropwise to the above solution, then add glyceryl monooleate, vortex and mix evenly, and then Add PEG 400, glyceryl dioleate and Gelucire 43/01 to the mixture respectively, and vortex and mix at room temperature to obtain hydroxycamptothecin cubic liquid crystal precursor composition B.

Example 3

A cubic liquid crystal precursor composition of hydroxycamptothecin, prepared from the following raw materials:

The cubic liquid crystal precursor composition of above-mentioned hydroxycamptothecin is prepared by the following method:

Weigh hydroxycamptothecin and dispersion medium N-methylpyrrolidone according to the above formula, disperse and dissolve hydroxycamptothecin in N-methylpyrrolidone, slowly add water to the above solution, then add phytantriol, vortex Then, PEG 400, glyceryl dioleate and Gelucire33/01 were added to the mixture, and the hydroxycamptothecin cubic liquid crystal precursor composition C was obtained after vortex mixing at room temperature.

Example 4

A cubic liquid crystal precursor composition of hydroxycamptothecin, prepared from the following raw materials:

The cubic liquid crystal precursor composition of above-mentioned hydroxycamptothecin is prepared by the following method:

Weigh hydroxycamptothecin and dispersion medium 2-pyrrolidone according to the above formula, disperse and dissolve hydroxycamptothecin in 2-pyrrolidone, slowly add water to the above solution, then add glycerol monooleate, and vortex to mix evenly , Then PEG 400, glyceryl dioleate and Gelucire39/01 were added to the mixture, and the hydroxycamptothecin cubic liquid crystal precursor composition D was obtained after vortex mixing at room temperature.

Example 5

A cubic liquid crystal precursor composition of hydroxycamptothecin, prepared from the following raw materials:

The cubic liquid crystal precursor composition of above-mentioned hydroxycamptothecin is prepared by the following method:

Weigh hydroxycamptothecin and the dispersion medium absolute ethanol according to the above formula, disperse and dissolve hydroxycamptothecin in absolute ethanol, slowly add water dropwise to the above solution, then add phytantriol, vortex and mix evenly, Then, PEG 400, glyceryl dioleate, Gelucire39/01 and Gelucire43/01 were added to the mixture, and the hydroxycamptothecin cubic liquid crystal precursor composition E was obtained after vortex mixing at room temperature.

Example 6

A cubic liquid crystal precursor composition of hydroxycamptothecin, prepared from the following raw materials:

The cubic liquid crystal precursor composition of above-mentioned hydroxycamptothecin is prepared by the following method:

Weigh hydroxycamptothecin and propylene glycol as the dispersion medium according to the above formula, disperse and dissolve hydroxycamptothecin in propylene glycol, slowly add water dropwise to the above solution, then add glyceryl monooleate, vortex and mix evenly, and then Add PEG 400, glyceryl dioleate and Gelucire33/01 to the mixture respectively, and vortex and mix at room temperature to obtain the cubic liquid crystal precursor composition F of hydroxycamptothecin.

Example 7

A cubic liquid crystal precursor composition of hydroxycamptothecin, prepared from the following raw materials:

The cubic liquid crystal precursor composition of above-mentioned hydroxycamptothecin is prepared by the following method:

Weigh hydroxycamptothecin and the dispersion medium absolute ethanol according to the above formula, disperse and dissolve hydroxycamptothecin in absolute ethanol, slowly add water dropwise to the above solution, then add phytantriol, vortex and mix evenly, Then, PEG 400, glyceryl dioleate, and Gelucire 43/01 were added to the mixture, and the hydroxycamptothecin cubic liquid crystal precursor composition G was obtained after vortex mixing at room temperature.

Example 8

A cubic liquid crystal precursor composition of hydroxycamptothecin, which is basically the same as the cubic liquid crystal precursor composition of hydroxycamptothecin in Example 1, the difference is that:

Among them, what PEG selects for use is PEG1000.

Referring to the method of Example 1, the cubic liquid crystal precursor composition H of hydroxycamptothecin was prepared.

Example 9

A cubic liquid crystal precursor composition of hydroxycamptothecin, which is basically the same as the cubic liquid crystal precursor composition of hydroxycamptothecin in Example 1, the difference is that:

Wherein the glyceryl oleate was selected from glyceryl trioleate.

Referring to the method of Example 1, the hydroxycamptothecin cubic liquid crystal precursor composition I was prepared.

Comparative example 1

A cubic liquid crystal precursor composition of hydroxycamptothecin, which is basically the same as the cubic liquid crystal precursor composition of hydroxycamptothecin in Example 1, the difference is that PEG, glyceryl oleate and glyceryl stearate are not added therein Esters, only with hydroxycamptothecin, liquid crystal material, water-miscible organic solvent and water.

Referring to the method and prescription in Example 1, the hydroxycamptothecin cubic liquid crystal precursor composition J was prepared.

Comparative example 2

A cubic liquid crystal precursor composition of hydroxycamptothecin, which is basically the same as the cubic liquid crystal precursor composition of hydroxycamptothecin in Example 1, the difference is that:

Wherein, what glyceryl stearate selects is Gelucire 33/01, and the consumption is 8 mg, namely 8%, and the consumption of liquid crystal material is 64 mg, namely 64%, and the rest components remain unchanged.

Referring to the method in Example 1, the hydroxycamptothecin cubic liquid crystal precursor composition K was prepared.

Cytotoxicity of Experimental Example 1 Hydroxycamptothecin Cubic Liquid Crystal Precursor Composition

The hydroxycamptothecin cubic liquid crystal precursor composition prepared in the above example 1 was subjected to a cytotoxicity experiment to investigate its toxic effect on liver cancer cells Hep G2 cells, and compare its cytotoxicity difference with that of hydroxycamptothecin injection, The experimental method is as follows: take the HepG2 cells in the logarithmic growth phase, dilute them with culture medium after digestion, count the cells and adjust to a single cell suspension of 1×10 ⁵ /ml. The above cell suspension was inoculated in a 96-well culture plate, 100 μl per well, 6 wells in each group. Place it in a 5% _CO2 incubator at 37°C for 24 hours. After the cells adhere to the wall, discard the culture solution, add different concentrations of hydroxycamptothecin cubic liquid crystal precursor composition and hydroxycamptothecin injection, and continue to place in 5 Incubate at 37°C for 24h in a %CO ₂ incubator. Cell viability was measured by MTT method, and the viability of tumor cells in each group was calculated.

The results are shown in Figure 1. The cytotoxicity of the hydroxycamptothecin cubic liquid crystal precursor composition prepared in Example 1 is significantly higher than that of the hydroxycamptothecin injection, and its toxicity ( _IC50 value is 0.5 μg/ml) is about 5 times of camptothecin injection (IC50 value is 2.5 μ g/ml), both compare, there is statistically significant difference (P<0.05), show that hydroxycamptothecin cubic liquid crystal precursor composition can improve The lethality of cancer cells.

Drug release characteristics of experimental example 2 hydroxycamptothecin cubic liquid crystal precursor composition

The hydroxycamptothecin cubic liquid crystal precursor compositions prepared in the above examples and comparative examples were investigated for drug release.

The experimental method is: accurately weigh an appropriate amount of hydroxycamptothecin cubic liquid crystal precursor composition and place it in a dialysis bag, seal it, put it into a centrifuge tube filled with 15ml pH7.4 phosphate buffer, and place it in a constant temperature shaker. Shake and incubate at a constant temperature of 37±0.5°C. Samples were taken at different time points. The sample solution was filtered through a 0.45 μm microporous membrane, and the drug content of the subsequent filtrate was determined by HPLC.

The experimental results show that the release curves of compositions A-G are all relatively close, and the release curve of the composition A is used as a representative to draw a drug release curve, as shown in Figure 2. As can be seen from the experimental results, the hydroxycamptotheca prepared by the present invention The alkali cubic liquid crystal precursor composition can be continuously released for 30 days in the phosphate buffer solution, which greatly prolongs the action time of hydroxycamptothecin in the body.

But, wherein composition H and I in embodiment 8-9, because the PEG used is PEG1000 (embodiment 8), and oleic acid glyceride is triolein (embodiment 9), the hydrophobicity of the two Both are greater than PEG400 and diolein, resulting in the formation of reversed phase hexagonal liquid crystals in the in-situ process. The hydrophobic groups are on the outer surface and the hydrophilic groups are on the inner surface, which is just the opposite of cubic liquid crystals, so it shows a certain The phenomenon of sudden release and rapid release does not achieve the effect of sustained release. In addition, due to the high melting point of PEG1000, it is easy to form a eutectic with hydroxycamptothecin in the in situ process, which makes it difficult to release part of hydroxycamptothecin, and also causes a certain slow and incomplete release phenomenon.

In the composition J of Comparative Example 1, since no PEG, glyceryl oleate and glyceryl stearate were added, there were obvious phenomena of burst release, incomplete release and instability. In the composition K of Comparative Example 2, due to the inappropriate amount of glyceryl stearate, reverse phase hexagonal liquid crystals were formed, resulting in burst release and rapid release, which was not conducive to the improvement of bioavailability.

Stability of Experimental Example 3 Hydroxycamptothecin in Cubic Liquid Crystal Precursor Composition and Release Process

The stability of the hydroxycamptothecin cubic liquid crystal precursor composition prepared in the above examples and comparative examples during the release process was investigated.

The experimental method is: place the hydroxycamptothecin cubic liquid crystal precursor composition at room temperature for 0h, 1d, 15d, 30d, 60d, 90d and 180d, and perform a release experiment according to the method of Experimental Example 2, releasing 0, 1, 12, 24, 48, 96, 240, 360, 480, 600 and 720h of the hydroxycamptothecin cubic liquid crystal precursor composition, dissolved in methanol solution with pH 6.5 to a constant volume, then took 20 μl for HPLC analysis, the analysis chromatogram For the chromatographic peaks that appear, if the hydroxycamptothecin is unstable and ring-opening occurs, the ring-opening form chromatographic peaks will appear at 9-10min, as shown in Figure 3. If hydroxycamptothecin can remain stable in the cubic liquid crystal precursor composition and its release process, and both exist in the closed ring form, then no ring-opened chromatographic peak will appear at 9-10 min, as shown in Figure 4 . The results are shown in the table below.

The stability of table 1 hydroxycamptothecin

Note: "None" means that no ring-opened chromatographic peaks have been observed until the end of the placement experiment or release experiment.

From the above experimental results, it can be seen that the hydroxycamptothecin cubic liquid crystal precursor composition prepared in Examples 1-7 was placed for 180 days and released in the phosphate buffer solution for 720 hours, and the hydroxycamptothecin had no ring opening. But, composition H and I in wherein embodiment 8-9, because the PEG used is PEG1000 (embodiment 8), and olein is triolein (embodiment 9), respectively in 30d and 15d ( Placement stability), 240h and 48h (release stability) began to appear ring-opened hydroxycamptothecin.

And in comparative example 1, owing to not adding PEG, glyceryl oleate and glyceryl stearate composition, the hydroxycamptothecin of ring-opening occurs in 1d (placement stability) and 12h (release stability), comparative example 2 In the process, due to the unsuitable amount of glyceryl stearate, a reversed phase hexagonal liquid crystal is formed, and hydroxycamptothecin is loaded on the outer surface of the reversed phase hexagonal liquid crystal aggregate. Stability) when ring-opened hydroxycamptothecin occurs.

Crystal Phase Stability of Experimental Example 4 Cubic Liquid Crystal Precursor Composition During Release

The crystal phase stability of the hydroxycamptothecin cubic liquid crystal precursor composition prepared in the above examples and comparative examples during the release process was investigated.

The experimental method is: carry out the release test according to the method of Experimental Example 2, and stop the release test at 0, 1, 12, 24, 48, 96, 240, 360, 480, 600 and 720 hours respectively, take out the substances in the dialysis bag, absorb Dry its surface moisture, and use polarized light microscopy and small-angle X-ray scattering for crystal phase confirmation.

As shown in Figure 5 in the SAXS spectrum, there are four main scattering peaks, and the ratio of the scattering factors conforms to the typical characteristic peak structure of cubic liquid crystal: It shows that the composition exists in the form of cubic liquid crystals during the release process, and no phase transition occurs. If the result is shown in Figure 6, the ratio of scattering factors q ₁ : q ₂ : q ₃ conforms to The ratio of , indicating that the composition has undergone a liquid crystal form transition during the release process, forming a reversed phase hexagonal liquid crystal.

The experimental results show that each sample is a dark field under a polarizing microscope, and the compositions H and I in Examples 8-9, since the PEG used is PEG1000 (Example 8), the glyceryl oleate is triolein Esters (Example 9), other forms of liquid crystals appeared at 360h and 48h respectively, and the state of coexistence of cubic liquid crystals and reversed-phase hexagonal liquid crystals in Example 9 was shown under a polarizing microscope.

In Comparative Example 1, since no PEG, glyceryl oleate, and glyceryl stearate were added, a liquid crystal transition occurred within 96 hours. In Comparative Example 2, an inverse phase was formed due to the inappropriate amount of glyceryl stearate. Hexagonal liquid crystals, so that the inverse hexagonal liquid crystals appear directly in 12h, and the release rate is obviously faster than that of the examples.

In summary, the present invention utilizes the unique structure, bio-affinity and stability of cubic liquid crystals, combined with in-situ technology, to develop a cubic liquid crystal precursor solution, which can be used as a good drug carrier for hydroxycamptothecin, effectively protecting The active group of the drug is increased, the stability of the drug is increased, the efficacy of the drug is improved, and the targeted drug delivery of hydroxycamptothecin is realized.

The above-mentioned embodiments only express several implementation modes of the present invention, and the description thereof is relatively specific and detailed, but should not be construed as limiting the patent scope of the present invention. It should be pointed out that those skilled in the art can make several modifications and improvements without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent for the present invention should be based on the appended claims.

Claims (10)

1. a cubic liquid crystal precursor composition for hydroxy camptothecin, is characterized in that, the raw material primarily of following percentage by weight is prepared from:

Wherein: disperse medium is made up of the raw material of following percentage by weight:

The organic solvent 8-30% dissolved each other with water

Water 70-92%.

2. the cubic liquid crystal precursor composition of hydroxy camptothecin according to claim 1, is characterized in that, described olein is glyceryl dioleate.

3. the cubic liquid crystal precursor composition of hydroxy camptothecin according to claim 1, is characterized in that, described PEG is PEG400.

4. the cubic liquid crystal precursor composition of hydroxy camptothecin according to claim 1, it is characterized in that, described tristerin is selected from: HLB value be 1 melting range be at least one in the Polyethylene Glycol tristerin of 33 DEG C-43 DEG C, and the percentage by weight of described tristerin is 2-4%.

5. the cubic liquid crystal precursor composition of hydroxy camptothecin according to claim 1, is characterized in that, the described organic solvent dissolved each other with water is selected from: at least one in propylene glycol, N-Methyl pyrrolidone, 2-Pyrrolidone and dehydrated alcohol.

6. the cubic liquid crystal precursor composition of the hydroxy camptothecin according to any one of claim 1-5, is characterized in that, described liquid crystal material is phytantriol and/or glyceryl monooleate.

7. the preparation method of the cubic liquid crystal precursor composition of the hydroxy camptothecin described in any one of claim 1-6, it is characterized in that, comprise the following steps: by recipe quantity, hydroxy camptothecin is dissolved in disperse medium, add mix homogeneously after liquid crystal material, add PEG, olein and tristerin respectively again, mix homogeneously, obtains the cubic liquid crystal precursor composition of hydroxy camptothecin.

8. the cubic liquid crystal precursor composition of the hydroxy camptothecin described in any one of claim 1-6 is preparing the application in hydroxy camptothecin cubic liquid crystal preparation.

9. the cubic liquid crystal precursor composition of hydroxy camptothecin according to claim 8 is preparing the application in hydroxy camptothecin cubic liquid crystal preparation, it is characterized in that, described hydroxy camptothecin cubic liquid crystal preparation is injection.

10. the cubic liquid crystal precursor composition of hydroxy camptothecin according to claim 9 is preparing the application in hydroxy camptothecin cubic liquid crystal preparation, it is characterized in that, the administering mode of described injection is in tumor body and/or tumor body sidenote is penetrated.