CN110664784A - Composite nano drug delivery system and application thereof in gynecological tumor treatment - Google Patents

Composite nano drug delivery system and application thereof in gynecological tumor treatment Download PDF

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CN110664784A
CN110664784A CN201911058234.0A CN201911058234A CN110664784A CN 110664784 A CN110664784 A CN 110664784A CN 201911058234 A CN201911058234 A CN 201911058234A CN 110664784 A CN110664784 A CN 110664784A
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mirna
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孙强
张文艳
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Hubei Green Unbounded Biotechnology Co ltd
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Abstract

The invention discloses a composite nano drug delivery system and application thereof in gynecological tumor treatment, comprising the following steps: (a) paclitaxel and curcumin composition as active ingredients; (b) miRNA-HA as a targeting material; (c) carboxylated magnetic mesoporous silica is used as a carrier. The invention uses the carboxylated magnetic mesoporous silicon dioxide as a drug carrier, on one hand, the larger drug loading capacity is ensured, and on the other hand, a modifiable point is provided for the modification of functional groups; by modifying the targeting material miRNA-HA, the targeting property of tumor cells of the carboxylated magnetic mesoporous silica nano drug delivery system and the targeting inhibition on gynecological tumor cells can be increased, and the inhibition rate on the migration and invasion of the tumor cells is improved. Meanwhile, the method has reasonable design, simple preparation process and wide application prospect, and lays a foundation for the design and development of a corresponding drug delivery system.

Description

Composite nano drug delivery system and application thereof in gynecological tumor treatment
Technical Field
The invention relates to the technical field of biological medicines, in particular to a composite nano drug delivery system and application thereof in gynecological tumor treatment.
Background
Ovarian cancer is one of the most common gynecological tumors, with the incidence rate second to cervical and uterine body cancer. Ovarian cancer has extremely high morbidity, and the mortality rate is higher than that of malignant tumors of female reproductive systems, so that the health of the vast female and the happiness of countless families are seriously affected: at present, the incidence rate of ovarian cancer in China is 8.14/10 ten thousand, and the death rate is 3.13/10 ten thousand. Ovarian cancer is not easy to be diagnosed in early stage, so about 70% of patients have been diagnosed as late stage at the first visit, which makes effective treatment of diseases face huge challenges, and according to treatment experience at home and abroad, the five-year survival rate of ovarian cancer patients is only 20% -30%, and has not been improved significantly in recent years.
Surgery and chemotherapy are common means for treating tumors, and the toxic and side effects of chemotherapeutic drugs are common problems in clinic, so that the life quality of patients is reduced, and finally, the treatment has to be abandoned frequently. Therefore, the toxic and side effects of chemotherapy are solved while the tumor is treated, and the improvement of the life quality becomes a hot spot of the current research.
The nano technology is applied to medical research in recent decades, and primary achievements are obtained. Liposomes are a nano-delivery system, and because their composition and structure are similar to cell membranes, their biocompatibility has unique advantages over synthetic materials. And because the preparation process is simple, the materials are easy to obtain (for example, soybean phospholipid has a pharmaceutical adjuvant batch for injection), and part of dosage forms enter industrial production, but the liposome has the defect of easy leakage, so that the stability of the liposome is reduced, the drug-loading rate is reduced, and the treatment effect of the horse liposome is finally reduced.
Therefore, there is a need in the art to develop a drug delivery system that can reduce the toxicity of antitumor drugs and improve antitumor effects and patient compliance.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provides a composite nano drug delivery system and application thereof in gynecological tumor treatment.
In order to achieve the purpose, the invention adopts the technical scheme that:
a composite nano drug delivery system comprising: (a) paclitaxel and curcumin composition as active ingredients; (b) miRNA-HA as a targeting material; (c) carboxylated magnetic mesoporous silica as a carrier;
the carboxylated magnetic mesoporous silica nano drug delivery system has active targeting combination and targeting inhibition capability on tumor cells, and can effectively inhibit migration and invasion of the tumor cells; targeting the miRNA in the material as a generation targeting inhibitor; hyaluronic Acid (HA) in the targeting material is used as a target molecule for active targeting delivery, the specific binding of the Hyaluronic Acid (HA) and a CD44 receptor on the surface of a tumor cell is utilized to realize the targeting identification of the tumor cell, and meanwhile, the HA is also used as a coupling molecule between miRNA and the carboxylated magnetic mesoporous silica.
As a further limitation of the above scheme, the preparation method of the carboxylated magnetic mesoporous silica comprises the following steps:
a) magnetic core (Fe)3O4) The preparation of (1):
using a 250mL three-necked flask, 120mL of distilled water was added, nitrogen gas was introduced, and 4.81g of FeCl was further added to the flask3.6H2O and 2.12gFeC12.4H2O, mechanically stirring, heating to 65 ℃, adding 10mL of 28% ammonia water solution, adding 0.85mL of oleic acid, reacting for 3h, washing with water to be neutral, and washing with absolute ethyl alcohol for two to three times to obtain Fe3O4Nanoparticles obtained byTo Fe3O4Dispersing the nano particles in chloroform, and performing ultrasonic treatment for about 30min until the nano particles are dissolved to obtain Fe3O4The content of (A) is 0.1 g/mL;
b) preparation of magnetic mesoporous silica:
dispersing 1.0g of CTAB in 50mL of aqueous solution, adding 2mL of chloroform dispersion of ferroferric oxide after complete dissolution, ultrasonically dispersing uniformly, heating to 60-70 ℃, and completely evaporating chloroform to obtain Fe3O4Adding 350mL of water and 2.8mL of NaOH (2M) into the aqueous dispersion, stirring to mix uniformly, heating to 35-40 ℃, adding 5mL of TEOS and 20mL of ethyl acetate, reacting for 6-10 h, washing with water for three times, and washing with absolute ethyl alcohol for three times to obtain Fe3O4/mSiO2Nanoparticles; then the obtained Fe3O4/mSiO2Nanoparticles were dispersed in 400mL of NH 4.0g3NO3Refluxing the obtained product in a 95% ethanol solution at 70-90 ℃ for 6-10 h, washing the obtained product with ethanol and then with water, and drying the product at 40 ℃ under a vacuum condition to obtain magnetic mesoporous silica for later use;
c) preparation of aminated magnetic mesoporous silica:
mixing 1g of magnetic mesoporous silica with 250mL of anhydrous toluene, ultrasonically dispersing uniformly, adding 20mLAPTES, stirring and reacting at 80 ℃ for 12h, washing twice with toluene and twice with anhydrous methanol, finally centrifuging at 8000rpm for 10min, and vacuum drying to obtain aminated magnetic mesoporous silica;
d) preparation of carboxylated magnetic mesoporous silica:
dispersing 10mg of aminated magnetic mesoporous silica in 20mL of DMSO solution, adding 15mg of succinic anhydride and 10mg of triethylamine, stirring and reacting for 24 hours at 60 ℃, washing the obtained product with ethanol and then with water, and drying in vacuum to obtain carboxylated magnetic mesoporous silica.
As a further limitation of the scheme, the concentration of the taxol in the active ingredients is 0.05-10 mg/mL, and the concentration of the curcumin is 20-80 ng/mL.
As a further limitation of the scheme, the concentration of the targeting material is 10-30 mg/mL.
As a further limitation of the scheme, the mass ratio of the targeting material to the carboxylated magnetic mesoporous silica is 1: 0.5-2.
As a further limitation of the above protocol, the molecular weight of the sodium hyaluronate is 37 KDa.
As a further limitation of the above protocol, the miRNA is miR-124.
The invention also provides a preparation method of the composite nano drug delivery system, which comprises the following steps:
s1 synthetic targeting material miRNA-HA
S11) dissolving sodium hyaluronate in deionized water, and swelling at room temperature for 12-24 h to obtain a sodium hyaluronate aqueous solution; the concentration of the sodium hyaluronate aqueous solution is 10-30 mg/mL;
s12) sequentially dripping 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) into a sodium hyaluronate aqueous solution to activate carboxyl in the sodium hyaluronate for 1-2 h to obtain a mixed solution; the mole ratio of EDC to NHS is 1: 1-0.5, wherein the molar ratio of carboxyl in the sodium hyaluronate to EDC is 1:0.5 to 2;
s13) adding miRNA into the mixed solution, and reacting for 24-48 h at room temperature under stirring; transferring the reaction solution into a dialysis bag after the reaction is finished, clamping two ends of the reaction solution, dialyzing and purifying by deionized water, and freeze-drying the solution in the dialysis bag to obtain a product miRNA-HA; the molar ratio of carboxyl in the sodium hyaluronate to miRNA is 10-5: 1;
s2 synthetic composite nano drug delivery system
S21) dissolving the target material miRNA-HA in deionized water, and swelling for 12-24 h at room temperature;
s22) sequentially dripping EDC and NHS into the solution obtained in the step S21), activating for 1-2 h, dripping carboxylated magnetic mesoporous silica into the solution, and stirring for 2-4 h at room temperature to obtain a reaction solution; the EDC/NHS molar ratio is 1: 1-0.5, wherein the molar ratio of carboxyl on the miRNA-HA to EDC is 1: 1-2;
s23), centrifuging the reaction solution, washing with deionized water, purifying, and freeze-drying to obtain the composite nano drug delivery system.
As a further limitation of the scheme, the cut-off molecular weight of the dialysis bag is 3500-375000, and the dialysis purification time is 24-48 h.
The invention also provides application of the composite nano drug delivery system in gynecological tumor treatment, and the composite nano drug delivery system is used for preparing drugs for treating gynecological tumors.
As a further limitation of the above, the tumor is ovarian cancer, breast cancer, uterine cancer, or a combination thereof.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention provides a composite nano drug delivery system, which utilizes carboxylated magnetic mesoporous silica as a drug carrier, on one hand, ensures larger drug loading capacity, and on the other hand, provides a modifiable point for modification of functional groups; by modifying the targeting material miRNA-HA, the targeting property of tumor cells of the carboxylated magnetic mesoporous silica nano drug delivery system and the targeting inhibition on gynecological tumor cells can be increased, and the inhibition rate on the migration and invasion of the tumor cells is improved.
(2) The invention develops a composite nano drug delivery system which has magnetic targeting effect and takes mesoporous silicon dioxide which is modified by function as a carrier material to wrap anti-tumor active substances and can generate anti-tumor substances in cooperation with the carrier material for the first time through extensive and intensive research. The composite nano drug delivery system developed by the invention comprises paclitaxel, curcumin, targeted inhibitory miRNA (especially miRNA for targeted inhibition of Ets-1 expression) and mesoporous silica with magnetic targeting effect and modified functions, wherein the paclitaxel, curcumin and miRNA synergistically generate an anti-tumor effect.
(3) The composite nano drug delivery system prepared by the invention can exert the synergistic anticancer effect of the drugs, has the functions of delivering active substances, treating tumors, reversing or reducing drug resistance and the like, passively targets tumor cells, is a biodegradable, sustained and controlled release and passively targeted carrier material, and is suitable for delivering the antitumor active substances.
(4) The method has reasonable design, simple preparation process and wide application prospect, and lays a foundation for the design and development of a corresponding drug delivery system.
Drawings
FIG. 1 shows the toxicity test results of the composite nano drug delivery system prepared in examples 1-5 of the present invention on human ovarian cancer SKOV-3 cells.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more clearly apparent, the present invention is further described in detail with reference to the following embodiments; it should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention; reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
The present invention is described in further detail below with reference to specific embodiments.
Additional and/or auxiliary additional active ingredients that inhibit tumor growth may also be included in the composite nanodrug delivery systems of the present invention.
In the medicine, the carboxylated magnetic mesoporous silica nano-drug delivery system has active targeting combination and targeting inhibition capability on tumor cells, and can effectively inhibit migration and invasion of the tumor cells; targeting miRNA in the material as a targeted inhibitor; hyaluronic Acid (HA) in the targeting material is used as a target molecule for active targeting delivery, the specific binding of the Hyaluronic Acid (HA) and a CD44 receptor on the surface of a tumor cell is utilized to realize the targeting identification of the tumor cell, and meanwhile, the HA is also used as a coupling molecule between miRNA and the carboxylated magnetic mesoporous silica. The paclitaxel, the curcumin and the targeted inhibitory miRNA are adopted as main active ingredients, and the paclitaxel, the curcumin and the targeted inhibitory miRNA can synergistically generate an anti-tumor effect, enhance the anti-tumor effect, weaken toxic and side effects and reduce the drug resistance.
In the present invention, paclitaxel, one of the first active ingredients, is a drug having anti-tumor activity, and is effective for cancers with high incidence, such as ovarian cancer, uterine cancer and breast cancer, and it can bind to specific sites of tubulin, promote the polymerization of microtubules, and stabilize them, thereby interfering with the functions of microtubules, such as cell motility, intracellular transport and mitosis, and thus inhibiting the growth of tumor cells. Paclitaxel, as a non-specific antitumor drug, has a strong killing effect on normal cells while killing tumor cells, thereby causing many side effects, such as nausea, vomiting, alopecia, and general soreness, and causing the quality of life of patients to be reduced. Curcumin is a natural effective component extracted from dried rhizome of Curcuma longa, Curcuma aromatica, and Curcuma zedoaria of Zingiberaceae, and has wide pharmacological action, low toxicity, and good tolerance. Curcumin has been shown to induce apoptosis, inhibit signal pathways, resist angiogenesis, and exert antitumor effects. The invention selects the curcumin and the paclitaxel to be combined as the main active ingredient for resisting the tumor, can further improve the synergistic treatment effect and reduce the toxic and side effect.
In addition to paclitaxel, the first active ingredient also includes analogs or derivatives of paclitaxel, such as docetaxel.
In the present invention, the other major active ingredient is a targeted inhibitory miRNA (as the second active ingredient).
Micro RNA (microRNA) is a kind of single-stranded small molecule non-coding RNA, has a length of about 22 nucleotides, widely exists in eukaryotes, and mainly causes degradation or post-transcriptional translation inhibition of target mRNA by complete or partial complementary binding with the untranslated region of the target mRNA 3', thereby regulating and controlling the expression of target genes.
The inventor discovers that the targeted inhibitory miRNA and the paclitaxel have excellent synergistic effect through a large amount of screening.
miR-124 is a micro RNA (microRNA) related to breast cancer apoptosis. Research reports that miR-124 can inhibit the proliferation of breast cancer MCF-7 and MDA-MB-231 cells by targeting Ets-1. Can inhibit the proliferation of tumor cells by inhibiting cyclin, has the function of regulating cell adhesion molecules, and has the problems of larger molecular weight, difficult cell uptake and in-vivo stability of miRNA compared with common medicines. Therefore, the miRNA is combined with hyaluronic acid to be used as a targeting material, Hyaluronic Acid (HA) is used as a target molecule for active targeting delivery, and the specific combination of the target molecule and a CD44 receptor on the surface of a tumor cell is utilized to realize the targeting recognition of the tumor cell so as to improve the synergistic treatment effect.
The growth of tumor cells is synergistically inhibited by the paclitaxel and the targeted inhibitory miRNA through different action mechanisms, the anti-tumor effect is improved, and simultaneously compared with the single administration of the paclitaxel and the targeted inhibitory miRNA, the synergistic administration of the paclitaxel and the targeted inhibitory miRNA can respectively reduce the administration dosage of the paclitaxel and the targeted inhibitory miRNA, so that the side effect is reduced, and the compliance of patients is improved.
The mode of administration of the composite nano drug delivery system of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): topical administration, peritumoral, intratumoral, intraperitoneal, intravenous, oral, topical, etc.
A preferred method is to administer the composite nano drug delivery system by injection, such as intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, intratumoral injection, paratumoral injection, transdermal administration, postoperative administration, etc.
The pharmaceutical preparation should be compatible with the mode of administration. The pharmaceutical composition of the present invention is preferably prepared in the form of an injection, for example, by a conventional method using physiological saline or an aqueous solution containing glucose and other adjuvants. Pharmaceutical compositions such as injections, solutions are preferably manufactured under sterile conditions. The amount of active ingredient administered is a therapeutically effective amount, for example from about 1 microgram per kilogram of body weight to about 5 milligrams per kilogram of body weight per day. In addition, the agents of the invention may be used (including before, during or after) with other therapeutic agents, such as antineoplastic agents.
In the case of pharmaceutical compositions, a safe and effective amount of the drug is administered to the mammal, wherein the safe and effective amount is generally at least about 10 micrograms/kg body weight, and in most cases no more than about 8 milligrams/dry gram body weight, preferably the dose is from about 10 micrograms/kg body weight to about 1 milligram/kg body weight. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.
In the present invention, the composite nano-drug delivery system is suitable for preparing a drug for treating cancer, especially for treating ovarian cancer, breast cancer, etc. and cancer recurred after operation, and in a preferred embodiment of the present invention, the cancer is selected from the following group: ovarian cancer, breast cancer, uterine cancer, or a combination thereof.
Example 1
The present embodiment provides a composite nano drug delivery system, comprising: (a) paclitaxel and curcumin composition as active ingredients; (b) miRNA-HA as a targeting material; (c) carboxylated magnetic mesoporous silica as a carrier;
the concentration of paclitaxel in the active ingredient is 5mg/mL, and the concentration of curcumin is 50 ng/mL; the concentration of the targeting material is 20 mg/mL; the mass ratio of the targeting material to the carboxylated magnetic mesoporous silica is 1: 1; the molecular weight of the sodium hyaluronate is 37 KDa; the miRNA is miR-124, and the sequence of the miR-124 is uaaggcacgcggugaaugcc.
The preparation method of the carboxylated magnetic mesoporous silica comprises the following steps:
a) magnetic core (Fe)3O4) The preparation of (1):
using a 250mL three-necked flask, 120mL of distilled water was added, nitrogen gas was introduced, and 4.81g of FeCl was further added to the flask3.6H2O and 2.12gFeC12.4H2O, mechanically stirring, heating to 65 ℃, adding 10mL of 28% ammonia water solution, adding 0.85mL of oleic acid, reacting for 3h, washing with water to be neutral, and washing with absolute ethyl alcohol for two to three times to obtain Fe3O4Nanoparticles of Fe obtained3O4Dispersing the nano particles in chloroform, and performing ultrasonic treatment for about 30min until the nano particles are dissolved to obtain Fe3O4The content of (A) is 0.1 g/mL;
b) preparation of magnetic mesoporous silica:
dispersing 1.0g CTAB in 50mL of aqueous solution, adding 2mL of ferroferric oxide chloroform dispersion after complete dissolution, ultrasonically dispersing uniformly, heating to 65 ℃ to evaporate chloroform completely to obtain Fe3O4Adding 350mL of water and 2.8mL of NaOH (2M) into the aqueous dispersion, stirring to uniformly mix the mixture, heating to 35-40 ℃, adding 5mL of TEOS and 20mL of ethyl acetate, reacting for 8 hours, respectively washing with water for three times, and washing with absolute ethyl alcohol for three times to obtain Fe3O4/mSiO2Nanoparticles; then the obtained Fe3O4/mSiO2Nanoparticles were dispersed in 400mL of NH 4.0g3NO3Refluxing the obtained product for 8 hours at 80 ℃ in the 95% ethanol solution, washing the obtained product with ethanol and then with water, and drying the product at 40 ℃ under a vacuum condition to obtain magnetic mesoporous silica for later use;
c) preparation of aminated magnetic mesoporous silica:
mixing 1g of magnetic mesoporous silica with 250mL of anhydrous toluene, ultrasonically dispersing uniformly, adding 20mLAPTES, stirring and reacting at 80 ℃ for 12h, washing twice with toluene and twice with anhydrous methanol, finally centrifuging at 8000rpm for 10min, and vacuum drying to obtain aminated magnetic mesoporous silica;
d) preparation of carboxylated magnetic mesoporous silica:
dispersing 10mg of aminated magnetic mesoporous silica in 20mL of DMSO solution, adding 15mg of succinic anhydride and 10mg of triethylamine, stirring and reacting for 24 hours at 60 ℃, washing the obtained product with ethanol and then with water, and drying in vacuum to obtain carboxylated magnetic mesoporous silica.
The preparation method of the composite nano drug delivery system comprises the following steps:
s1 synthetic targeting material miRNA-HA
S11) dissolving sodium hyaluronate in deionized water, and swelling for 18h at room temperature to obtain sodium hyaluronate aqueous solution; the concentration of the sodium hyaluronate aqueous solution is 10-30 mg/mL;
s12) sequentially dripping 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) into the sodium hyaluronate aqueous solution to activate carboxyl in the sodium hyaluronate for 1.5h to obtain a mixed solution; the mole ratio of EDC to NHS is 1:1, the molar ratio of carboxyl groups to EDC in the sodium hyaluronate is 1: 1;
s13) adding miRNA into the mixed solution, and reacting for 36h at room temperature under stirring; transferring the reaction solution into a dialysis bag with the molecular weight cutoff of 3500 after the reaction is finished, clamping two ends of the reaction solution, dialyzing and purifying the reaction solution with deionized water for 36 hours, and freeze-drying the solution in the dialysis bag to obtain a product miRNA-HA; the molar ratio of carboxyl in the sodium hyaluronate to miRNA is 8: 1;
s2 synthetic composite nano drug delivery system
S21) dissolving the target material miRNA-HA in deionized water, and swelling for 18h at room temperature;
s22) adding EDC and NHS dropwise into the solution obtained in the step S21) in sequence, activating for 1h, adding carboxylated magnetic mesoporous silica dropwise into the solution, and stirring for 3h at room temperature to obtain a reaction solution; the EDC/NHS molar ratio is 1:1, the molar ratio of carboxyl on the targeting material miRNA-HA to EDC is 1: 1.5;
s23) centrifuging the reaction solution at 8000r/min, washing with deionized water, purifying, and lyophilizing to obtain the compound nanometer drug delivery system.
The average particle size of the composite nano drug delivery system prepared by the embodiment is 125.9nm, the polydispersity is 0.168, the encapsulation efficiency is more than 97%, the drug-loading rate of paclitaxel is 3.752%, the drug-loading rate of curcumin is 3.148%, and the drug-loading rate of miR-124 is 0.1476 nmol/mg.
Example 2
This example provides a composite nano drug delivery system, which differs from example 1 in that: only paclitaxel is used as an active ingredient, and the concentration of the paclitaxel in the active ingredient is 5 mg/mL.
The rest is the same as in example 1.
Example 3
This example provides a composite nano drug delivery system, which differs from example 1 in that: curcumin is only used as an active ingredient, and the concentration of the curcumin in the active ingredient is 50 ng/mL.
The rest is the same as in example 1.
Example 4
This example provides a composite nano drug delivery system, which differs from example 1 in that: the targeting material is HA.
The rest is the same as in example 1.
Example 5
This example provides a composite nano drug delivery system, which differs from example 1 in that: the carrier is aminated magnetic mesoporous silica.
The rest is the same as in example 1.
Test examples
In vitro inhibition experiments
The MTT method is adopted to detect the inhibition effect of the composite nano drug delivery system prepared in the embodiment 1-5 on the proliferation of ovarian cancer cell SKOV-3 cells. And (3) inoculating SKOV-3 cells in a logarithmic growth phase to a 96-well culture plate, and respectively adding 5 preparations prepared in the examples 1-5 after adherence, wherein the concentrations of paclitaxel and curcumin in each group are kept consistent and are respectively 5mg/mL and 50 ng/mL. The blank control group was incubated for 24/48 hours with medium (control) containing no drug, and the medium containing drug was aspirated for MTT assay.
The results are shown in fig. 1, where the nanoparticle group (example) co-loaded with paclitaxel and curcumin at a relative increment rate of 48 hours is the smallest, significantly smaller than the two single drug groups.
The above results indicate that in the present invention, curcumin as an active ingredient can produce a synergistic effect when used in combination with paclitaxel. The combined use has obviously better inhibition effect on tumor cells than the single use. The nano drug delivery system is formed by adopting the carboxylated magnetic mesoporous silica as the carrier, has active targeting combination and targeting inhibition capability on tumor cells, and can effectively inhibit the migration and invasion of the tumor cells; meanwhile, miRNA in the targeting material can be used as a targeting inhibitor; hyaluronic Acid (HA) in the targeting material is used as a target molecule for active targeting delivery, the specific binding of the Hyaluronic Acid (HA) and a CD44 receptor on the surface of a tumor cell is utilized to realize the targeting identification of the tumor cell, and meanwhile, the HA is also used as a coupling molecule between miRNA and the carboxylated magnetic mesoporous silica. By adopting the paclitaxel, the curcumin and the targeted inhibitory miRNA as main active ingredients, the paclitaxel, the curcumin and the targeted inhibitory miRNA can synergistically generate an anti-tumor effect, enhance the anti-tumor effect, weaken toxic and side effects and reduce the drug resistance.
While the invention has been described with respect to specific embodiments thereof, it will be understood by those skilled in the art that the foregoing and other changes, omissions and deviations in the form and detail thereof may be made without departing from the scope of this invention; those skilled in the art should appreciate that they can readily use the disclosed conception and specific embodiments as a basis for designing or modifying other structures for carrying out the same purposes of the present invention without departing from the spirit and scope of the invention; meanwhile, any equivalent changes, modifications and alterations of the above embodiments according to the spirit and techniques of the present invention are also within the scope of the present invention.

Claims (10)

1. A composite nano drug delivery system, comprising: (a) paclitaxel and curcumin composition as active ingredients; (b) miRNA-HA as a targeting material; (c) carboxylated magnetic mesoporous silica as a carrier;
the carboxylated magnetic mesoporous silica nano drug delivery system has active targeting combination and targeting inhibition capability on tumor cells, and can effectively inhibit migration and invasion of the tumor cells; targeting the miRNA in the material as a generation targeting inhibitor; hyaluronic Acid (HA) in the targeting material is used as a target molecule for active targeting delivery, the specific binding of the Hyaluronic Acid (HA) and a CD44 receptor on the surface of a tumor cell is utilized to realize the targeting identification of the tumor cell, and meanwhile, the HA is also used as a coupling molecule between miRNA and the carboxylated magnetic mesoporous silica.
2. The composite nano drug delivery system of claim 1, wherein the concentration of paclitaxel in the active ingredient is 0.05-10 mg/mL, and the concentration of curcumin is 20-80 ng/mL.
3. The composite nano drug delivery system according to claim 1, wherein the concentration of the targeting material is 10-30 mg/mL.
4. The composite nano drug delivery system according to claim 1, wherein the mass ratio of the targeting material to the carboxylated magnetic mesoporous silica is 1: 0.5-2.
5. The composite nano drug delivery system of claim 1, wherein the sodium hyaluronate has a molecular weight of 37 kDa.
6. The composite nano drug delivery system of claim 1, wherein the miRNA is miR-124.
7. The method for preparing a composite nano drug delivery system as claimed in any one of claims 1 to 6, comprising the steps of:
s1 synthetic targeting material miRNA-HA
S11) dissolving sodium hyaluronate in deionized water, and swelling at room temperature for 12-24 h to obtain a sodium hyaluronate aqueous solution; the concentration of the sodium hyaluronate aqueous solution is 10-30 mg/mL;
s12) sequentially dripping 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) into a sodium hyaluronate aqueous solution to activate carboxyl in the sodium hyaluronate for 1-2 h to obtain a mixed solution; the mole ratio of EDC to NHS is 1: 1-0.5, wherein the molar ratio of carboxyl in the sodium hyaluronate to EDC is 1:0.5 to 2;
s13) adding miRNA into the mixed solution, and reacting for 24-48 h at room temperature under stirring; transferring the reaction solution into a dialysis bag after the reaction is finished, clamping two ends of the reaction solution, dialyzing and purifying by deionized water, and freeze-drying the solution in the dialysis bag to obtain a product miRNA-HA; the molar ratio of carboxyl in the sodium hyaluronate to miRNA is 10-5: 1;
s2 synthetic composite nano drug delivery system
S21) dissolving the target material miRNA-HA in deionized water, and swelling for 12-24 h at room temperature;
s22) sequentially dripping EDC and NHS into the solution obtained in the step S21), activating for 1-2 h, dripping carboxylated magnetic mesoporous silica into the solution, and stirring for 2-4 h at room temperature to obtain a reaction solution; the EDC/NHS molar ratio is 1: 1-0.5, wherein the molar ratio of carboxyl on the miRNA-HA to EDC is 1: 1-2;
s23), centrifuging the reaction solution, washing with deionized water, purifying, and freeze-drying to obtain the composite nano drug delivery system.
8. The method for preparing a composite nano drug delivery system according to claim 7, wherein the cut-off molecular weight of the dialysis bag is 3500-375000, and the dialysis purification time is 24-48 h.
9. Use of a composite nano drug delivery system according to any one of claims 1 to 6 in the treatment of gynecological tumors, for the preparation of a medicament for the treatment of gynecological tumors.
10. Use of a composite nano drug delivery system according to claim 9 in the treatment of gynecological tumors, wherein said tumors are ovarian cancer, breast cancer, uterine cancer or a combination thereof.
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