CN104398501A - Polymeric micelle composition for treating recrudescent cancer and preparation method thereof - Google Patents

Abstract

The invention discloses a polymer micelle composition for treating recurrent cancer and a preparation method thereof. The composition comprises an amphiphilic block polymer and an active ingredient, wherein the active ingredient is taxane and Induction of tumor stem cell apoptosis promoter resveratrol. The polymer micelle composition used for treating cancer prone to relapse can accumulate in tumor tissue, kill common tumor cells and tumor stem cells simultaneously, improve the therapeutic effect of tumor and reduce the probability of tumor metastasis and recurrence.

Description

Polymer micelle composition for treating relapse-prone cancer and preparation method thereof

technical field

The invention relates to a polymer micelle composition for treating recurrent cancer and a preparation method thereof.

Background technique

Malignant tumors are common and frequently-occurring diseases that seriously threaten human life and health, and have become the leading cause of human death worldwide. Antitumor drugs represented by paclitaxel and docetaxel are widely used in the treatment of malignant tumors such as breast cancer, ovarian cancer, and non-small cell lung cancer. However, the solubility of taxane anticancer agents such as paclitaxel and docetaxel in water is very low, and non-ionic surfactants are usually used for solubilization. Paclitaxel injection Taxol (Taxol) is to dissolve the drug in a mixed solvent of polyoxyethylene castor oil and absolute ethanol (1:1). Docetaxel injection taxotere (Taxotere) is that the drug is dissolved in the surfactant Tween-80 (40g/L), then subpackaged, and then diluted with 13% ethanol aqueous solution and 5% glucose solution before clinical use. Surfactant polyoxyethylene castor oil releases histamine when it degrades in the body, which can cause severe allergic reactions and bone marrow suppression and other side effects. Surfactant Tween-80 is hemolytic and viscous, and can cause obvious allergies after injection reaction.

Although taxane has become the first-line drug for the clinical treatment of various malignant tumors, its anti-tumor effect has also been generally recognized. However, as we all know, the cure rate of tumor chemotherapy is extremely low, and one of the most important reasons is tumor recurrence after chemotherapy. Once tumor recurrence occurs, the current clinical treatment methods will basically not produce any therapeutic effect. Recent studies have shown that there is heterogeneity of cells in tumor tissues, including ordinary tumor cells and a small number of cancer stem cells. Conventional chemotherapy drugs, including taxanes, can only kill normal tumor cells, but not cancer stem cells. After chemotherapy, tumor recurrence and metastasis are caused by the existence of tumor stem cells. Therefore, tumor stem cells are considered to be the main reason for tumor proliferation, recurrence and metastasis. Because cancer stem cells do not respond to common chemotherapy, radiotherapy and other treatments, there is currently a lack of clinical treatment strategies for cancer stem cells.

Resveratrol is a polyphenolic substance extracted from plants. Studies have shown that it can induce tumor stem cell apoptosis. Resveratrol can induce tumor stem cell apoptosis by activating the P53 gene, resulting in proteasome-dependent cytokine degradation. Apoptosis occurs. However, the solubility of resveratrol in water is extremely low, and it is not easy to make intravenous injections. It is necessary to select suitable excipients to solubilize it to meet clinical needs.

On the other hand, the combined use of resveratrol and taxanes has a synergistic effect, which can reduce the dosage of taxanes, reduce the occurrence of toxic and side effects, and increase the therapeutic effect of tumors.

At present, there is no polymer micelle composition containing taxane and tumor stem cell apoptosis promoter resveratrol at the same time, and there is no micelle composition that uses two drugs to treat recurrent cancer.

Contents of the invention

technical problem

The purpose of the present invention is to provide a polymer micelle composition for the treatment of prone to relapse cancer, the composition comprises an amphiphilic block polymer and an active ingredient, wherein the active ingredient is taxane and tumor stem cell apoptosis Accelerator, the tumor stem cell apoptosis accelerator is resveratrol.

The present invention also aims to provide a preparation method of the polymer micelle composition for treating relapse-prone cancer.

The present invention is achieved through the following technical solutions:

The present invention provides a polymer micelle composition for treating cancer prone to relapse, the composition comprises an amphiphilic block polymer and an active ingredient, wherein the active ingredient is taxane and tumor stem cell apoptosis-promoting agent, the tumor stem cell apoptosis promoter is resveratrol.

Polymer micelle composition of the present invention comprises:

(1) a mixture of taxane and resveratrol encapsulated in a micelle;

(2) It can also be a mixture of encapsulated taxane polymer micelles and encapsulated resveratrol polymer micelles;

A mixture of the above (1) and (2) is also possible.

The mass ratio of the active ingredients taxane and resveratrol is: 0.1-10:1, preferably 0.5-2:1, the sum of taxane and resveratrol accounts for 0.1%-45wt% of the micellar composition .

The polymer micelle composition of the present invention comprises the following components by weight percentage:

Taxane 0.2-30%;

Resveratrol 0.2-20%;

Amphiphilic block copolymer 50-99.6%.

Wherein the taxane can be paclitaxel, docetaxel, 7-acetyl paclitaxel, t-acetyl paclitaxel, 10-deacetyl paclitaxel, 10-deacetyl-7-epitaxel, 7-xyloside paclitaxel , 10-deacetyl-7-glutaryl paclitaxel, 7-N,N-dimethylglycyl paclitaxel, 7-L-alanyl paclitaxel or a mixture of the above. In particular, it may be docetaxel or paclitaxel. The taxane can be amorphous or crystalline. Can be dehydrated or hydrated.

The amphiphilic block copolymer is a polymer comprising a hydrophilic block (A) and a hydrophobic block (B), wherein the hydrophilic block is selected from polyethylene glycol and methoxypolyethylene glycol One or more blocks of alcohol; the hydrophobic block is selected from one or more blocks of the following: polylactic acid, polylactide, polylactic acid glycolic acid, polyglycolic acid lactide, polymandelic acid, poly Caprolactone, polydioxanone, polyamino acid, polyorthoester, polyanhydride and copolymers of the above.

The number average molecular weight of the hydrophilic block is 500-20,000 Daltons, the number average molecular weight of the hydrophobic block is 500-20,000 Daltons, and the hydrophilic block is in the total The content in mass is 20-80wt%.

The resveratrol is used to induce tumor stem cell apoptosis, and can be in cis or trans structure. Resveratrol is a natural polyphenol compound, which was first extracted from resveratrol and exists in trees, peanuts, and grapes. Resveratrol has two isomers, cis and trans.

In one embodiment of the present invention, the polymer micelle composition for treating recurrent cancer includes taxane, resveratrol and two amphiphilic block copolymers solubilized by poorly water-soluble drugs.

In another embodiment, the polymer micelles are nanoparticles. In particular, the particle size of the micellar nanoparticles is 10-200 nm.

The term "cancer prone to relapse" here refers to the failure of anticancer agents such as taxanes to kill tumor stem cells, resulting in the continued differentiation of tumor stem cells into tumor cells after chemotherapy with anticancer agents, resulting in recurrence of controlled tumors or transfer. The cancer includes all types of cancer, such as: breast cancer, ovarian cancer, lung cancer, prostate cancer, etc.; without any limitation.

The term "nanoparticle" herein refers to particles having a nanoscale particle size, including micellar structured nanoparticles.

The expression "complex" used to describe "complex micelles", "composite nanoparticles", "composite micelles and nanoparticles", or such micelles means a state in which taxane and resveratrol Together, retrol is encapsulated in a micelle or nanoparticle. Composite micelles or nanoparticles can be obtained by encapsulating two drugs in micelles or nanoparticles.

The polymer micelle composition according to the invention includes mixed micelles and composite micelles and mixtures of taxanes and resveratrol encapsulated therein. In one embodiment, a micellar composition for treating relapse-prone cancer comprises active ingredients taxane and resveratrol encapsulated in a micelle consisting of an amphiphilic block copolymer . In particular, the composition may be in the form of a complex micelle or in a mixed form. If it is in the form of a complex micelle, the taxane and resveratrol are encapsulated together in the same micelle particle composed of an amphiphilic block copolymer In the mixed form, the taxane and resveratrol are encapsulated in different micellar particles and then mixed. In another embodiment, the composition may be a mixture of: a composition comprising taxane and resveratrol encapsulated together in the same micelle particle; and a taxane-encapsulating polymer micelle Or a composition encapsulating a resveratrol polymer micelle mixture.

In the anticancer agent composition according to the embodiment of the present invention, the mass ratio of taxane to resveratrol (mass of taxane:mass of resveratrol) may be 0.1-10:1. It is preferably 0.5-2:1. Within this range of mass ratio, the effect of taxane as an anticancer agent in treating and preventing tumor recurrence is optimal.

In another embodiment, based on the total mass of the composition, the total content of taxane and resveratrol may be 0.1-45 wt%. Since taxanes and resveratrol are encapsulated in polymer micelles, the content of taxanes and resveratrol that can be encapsulated in the core of polymer micelles is limited. In one embodiment of the present invention, based on the total mass of the composition, the content of taxane is 0.2-30wt%, preferably 0.2-20%; and the content of resveratrol is 0.2-15wt%, preferably 0.2-10 %.

In one embodiment, the amphiphilic block copolymer may be an A-B type diblock copolymer, including hydrophilic A and hydrophobic B. The hydrophilic block A can be polyethylene glycol, and the hydrophobic block B can be polylactic acid or its derivatives.

Polyethylene glycol as the hydrophilic block A can be selected from one or more of polyethylene glycol or methoxypolyethylene glycol, but is not limited to these polyethylene glycols. In particular, methoxypolyethylene glycol is preferred.

Polylactic acid or its derivatives as the hydrophobic block B can be one or more substances selected from the following: polylactic acid, polylactide, polyethanol, polyglycolide, polymandelic acid, polycaproic acid Lactone, polydioxanone, polyamino acid, polyorthoester, polyanhydride and copolymers of the above. Preferably, the polylactic acid or its derivatives may be polylactic acid, polylactide, polyglycolide, polymandelic acid, polycaprolactone, polydioxanone.

In one embodiment, the number average molecular weight of the hydrophilic block is 500-20,000 Daltons, preferably 1,000-10,000 Daltons. The number average molecular weight of the hydrophobic block B is 500-10,000 Daltons. In another embodiment, based on the total weight of the diblock copolymer, the content of the hydrophilic block A can be 20-80wt%, preferably 40%-70wt%.

The mass percentage content of the amphiphilic block copolymer in the total mass of the composition may be 50-99.5%, preferably 50-90%.

The composite amphiphilic block copolymer composition (taxane and resveratrol are encapsulated in micelles together) has a particle size of 10-180nm in water and can be solid after freeze-drying.

The invention also provides a preparation method of the micellar composition for treating recurrent cancer. According to one embodiment of the present invention, the preparation method of a polymer micelle composition containing taxane and resveratrol as active ingredients for treating recurrent cancer may include the following steps: (a) adding purple Dissolving the cane, resveratrol and amphiphilic block copolymer in the organic solvent; (b) mixing the organic solvent with the aqueous medium; (c) dividing the solution into glass containers and freeze-drying.

Taxane and resveratrol can also be dissolved in different organic solvents respectively with described polymer, thus make taxane-containing polymer micelle and resveratrol-containing polymer micelle, then these two The micelles are mixed to obtain a mixed micelles composition.

The micelle composition according to the present invention can be composite type or mixed type, if it is composite type, taxane and resveratrol are encapsulated in polymer micelles or nanoparticles together, And if it is a hybrid type, the taxane-containing polymer micelles or nanoparticles are mixed with the resveratrol-containing polymer micelles or nanoparticles. Preferably, the above two types of compositions can be prepared as follows.

A preparation method of a composite anticancer agent polymer micelle composition, wherein taxane and resveratrol are encapsulated in the micelle composition together as active ingredients, and the method may include the following steps :

(a) dissolving taxane, resveratrol and amphiphilic block copolymer in an organic solvent selected from the group consisting of: ethanol, methanol, propanol, butanol, pentanol, hexanol, acetone, Acetonitrile, dichloromethane, chloroform, methyl ethyl ketone, and ethyl acetate.

(b) mixing an organic solvent with an aqueous solution to prepare a mixture in which the taxane, resveratrol and polymer are homogeneously mixed;

(c) Divide the solution into glass containers and lyophilize.

After step (b) of the preparation method, the following steps may also be included:

(d) sterilizing and depyrogenating the resulting micellar composition;

In step (d), the aqueous solution is sterilized by filtering the solution through a sterilizing filter.

In one embodiment, the aqueous solution used in step (b) may be distilled water, physiological saline, or an aqueous solution of a lyophilized adjuvant. The freeze-drying adjuvant can be selected from one or more substances selected from mannitol, sorbitol, lactose, trehalose and sucrose, preferably lactose and trehalose.

A preparation method of a mixed anticancer agent polymer micelle composition, the micelle composition comprising a taxane-containing polymer micelle or nanoparticle and a resveratrol-containing polymer micelle or nanoparticle The mixture, described method comprises the following steps:

(a) taxane, resveratrol and amphiphilic block copolymer are dissolved in organic solvent respectively, and described organic solvent is selected from: ethanol, methanol, propanol butanol, pentanol, hexanol, acetone , acetonitrile, dichloromethane, chloroform, methyl ethyl ketone and ethyl acetate;

(b) Mix the obtained organic solution and aqueous solution to prepare taxane-containing polymer micelles and resveratrol-containing polymer micelles, and mix different micelles to prepare a mixed-type micellar composition.

(c) Divide the solution into glass containers and lyophilize.

The preparation method may also include other steps after step (c):

(d) Sterilizing and depyrogenating the resulting micellar composition.

In step (b), the aqueous solution used may be distilled water, physiological saline, or an aqueous solution of a lyophilized adjuvant. The freeze-drying adjuvant can be selected from one or more substances selected from mannitol, sorbitol, lactose, trehalose and sucrose, preferably lactose and trehalose.

In steps (b) and (c), the organic solution and the aqueous solution containing the two drugs can also be mixed separately, freeze-dried separately, and the micelles are mixed just before use.

The polymer micelle composition of the present invention can be prepared into a clinically acceptable preparation with a pharmaceutically acceptable carrier. Preparations for parenteral administration can be administered in the following ways: sublingual administration, rectal administration, transdermal administration, intravenous injection administration, intramuscular injection administration, intraperitoneal injection administration and the like. A typical example is an isotonic aqueous solution or suspension for intravenous injection. In one embodiment of the present invention, the micellar composition for treating recurrent cancer can be made into a freeze-dried preparation, and can be reconstituted in water for injection, 5% glucose solution, normal saline or the like, and passed Administered intravenously.

Since both taxanes and resveratrol are poorly soluble in water, specific techniques and compositions are required for solubilization for intravenous administration. The usual technical means is to solubilize such drugs with organic solvents (ethanol) or surfactants (polyoxyethylene castor oil or polysorbate). However, these two surfactants need to be used with caution because they can cause severe allergic reactions when administered intravenously. The present invention provides a polymer composition without obvious side effects (such as hemolysis and allergic reaction), even if it is administered in the form of intravenous injection, there is no obvious side effect.

The polymer micelle composition for the treatment of relapse-prone cancer according to the present invention comprises amphiphilic block copolymer, due to the enhanced permeability and retention (enhanced permeability and retention, EPR) effect, the polymer gel The beam composition can be deposited on tumor tissue in high concentrations. And because the composition contains taxanes and tumor stem cell apoptosis promoters, it exhibits excellent anticancer efficacy due to its killing of tumor stem cells. In addition, the composition of the present invention does not produce hemolytic and allergic reactions due to the use of polyoxyethylene castor oil. By using resveratrol in combination, the dosage of taxanes can be reduced, the occurrence of toxic reactions can be reduced, and the therapeutic effect of tumor can be enhanced.

Description of drawings

Fig. 1 is the particle size distribution result of the polymer micelle that packs docetaxel and resveratrol simultaneously, the particle size distribution figure of the described polymer micelle analyzed by dynamic light scattering method in aqueous solution;

Fig. 2 is the measurement result of the stability of polymer micelles loaded with docetaxel and resveratrol simultaneously, wherein, docetaxel and resveratrol are determined by high performance liquid chromatography (HPLC);

Figure 3 is the cytotoxic effect of docetaxel and resveratrol drug-loaded micelles on MCF-7 cells;

Figure 4 is the cytotoxic effect of docetaxel and resveratrol drug-loaded micelles on MCF-7 tumor stem cells;

Figure 5 shows the pharmacodynamic effects of docetaxel and resveratrol drug-loaded micelles on MCF-7 tumor-bearing mice.

Detailed ways

[Example 1] Preparation of a composite anticancer agent micelle composition containing an amphiphilic block copolymer, wherein docetaxel and resveratrol are encapsulated together in the micelle composition.

Preparation of compound anticancer agent composition for treating recurrent cancer, the anticancer agent composition includes monomethoxypolyethylene glycol-polylactide (mPEG-PDLLA), polyethylene glycol-polyaspartic acid amino acid (PEG-PASP), polyethylene glycol-polylactide-glycolide (mPEG-PLGA) amphiphilic block copolymer, taxane and resveratrol are encapsulated in polymer micelles together middle.

The substances in Table 1 are dissolved in 15.0ml of propanol, then mixed with water, in this mixed solution, docetaxel, resveratrol and mPEG-PDLLA polymer (monomethoxypolyethylene glycol weight average Molecular weight 2000, polylactide weight average molecular weight 2000, mPEG2000-PLA2000) uniformly mixed. Weigh 500mg of lactose and dissolve in 10ml of water for injection. Mix propanol and water evenly under agitation, filter the solution through a 0.22-micron filter membrane, and pack it into clean vials after sterilization. The vials are placed in the freezer at -30°C-50°C, freeze-dried, filled with nitrogen, stoppered, and capped to obtain the product.

【Table 1】

[Example 2] Preparation of a composite polymer nanoparticle composition simultaneously encapsulating docetaxel and resveratrol.

The material in table 2 is dissolved in 20ml amyl alcohol, then mixes with water, in this mixed solution, docetaxel, resveratrol and mPEG-PDLLA polymer (monomethoxypolyethylene glycol weight average molecular weight 2000 , polylactide weight average molecular weight 6000, MPEG2000-PLA6000) uniformly mixed. Weigh 500mg trehalose and dissolve it in 10ml water for injection. Mix propanol and water evenly under agitation, filter the solution through a 0.22-micron filter membrane, and pack it into clean vials after sterilization. The vials are placed in the freezer at -30°C-50°C, freeze-dried, filled with nitrogen, stoppered, and capped to obtain the product.

【Table 2】

[Example 3] Preparation of a mixed anticancer agent micelle composition, which includes encapsulating docetaxel-containing polymer micelles and encapsulating resveratrol-containing polymer micelles. After preparing polymer micelle compositions encapsulating docetaxel and resveratrol respectively, the two micelle compositions were mixed to prepare a micelle composition for treating relapse-prone cancer.

a) Preparation of amphiphilic block copolymer micelles encapsulating resveratrol

Take resveratrol 10.0mg and mPEG-PDLLA polymer (monomethoxypolyethylene glycol weight average molecular weight 2000, polylactide weight average molecular weight 2000, mPEG2000-PLA2000) 100mg, dissolve in 20ml butanol, then Mixed with water, in this mixture, resveratrol and mPEG-PDLLA polymer (monomethoxypolyethylene glycol weight average molecular weight 2000, polylactide weight average molecular weight 2000, mPEG2000-PLA2000) uniformly mixed, decompression The organic solvent was removed.

b) Preparation of an amphiphilic block copolymer encapsulating docetaxel

Take docetaxel 10.0mg and mPEG-PDLLA polymer (monomethoxypolyethylene glycol weight average molecular weight 2000, polylactide weight average molecular weight 2000, MPEG2000-PLA2000) 100mg, dissolve in 20ml butanol, then mix with Mixed with water, in this mixture, docetaxel and mPEG-PDLLA polymer (monomethoxypolyethylene glycol weight average molecular weight 2000, polylactide weight average molecular weight 2000, MPEG2000-PLA2000) are uniformly mixed, and removed under reduced pressure Organic solvents.

c) Preparation of Mixed Polymer Micellar Composition

Mix the above two solutions. Weigh 500 mg of trehalose and dissolve it in the solution. The solution is sterilized and filtered through a 0.22-micron filter membrane, and then sterilized and dispensed into clean vials. The vials are placed in the freezer at -30°C-50°C, freeze-dried, filled with nitrogen, stoppered, and capped to obtain the product.

d) Preparation of Mixed Polymer Micellar Composition

Adopt step a) and step and step b) to prepare the freeze-dried product of the lyophilized product of the resveratrol-loaded polymer micelle composition and the lyophilized product of the docetaxel polymer micelle composition respectively, the lyophilized product is in an aqueous medium Reconstitute and mix again.

The particle size of the prepared composition was measured. The results are shown in Table 4

【Table 4】

[Test Example 1] Particle size of composite micelles encapsulating docetaxel and resveratrol at the same time.

The compositions prepared in Examples 1 and 2 were redissolved in physiological saline to prepare samples for particle size determination. The particle size of the diluted composition solution was measured using a particle size analyzer. The results are shown in Table 5. A typical particle size distribution diagram is shown in Figure 1. It can be seen from the figure that the particle size distribution range of the drug-loaded micelles is narrow, and the polydispersity index is less than 0.1.

sample Particle size (nm) Composition 1 20-30 Composition 2 17-28 Composition 3 16-30 Composition 4 22-34 Composition 5 20-30 Composition 6 22-34 Composition 7 20-30

[Determination Example 2] Stability of Simultaneously Encapsulating Docetaxel and Resveratrol Micelles

Composition 4 diluted in Test Example 1 was left at room temperature for 24 hours to evaluate the stability of the solution. During the test period, the encapsulation efficiency of docetaxel and resveratrol was measured by high performance liquid chromatography (HPLC). The results are shown in Figure 2, and the specific HPLC conditions are as follows:

As shown in Figure 2, the concentrations of paclitaxel and resveratrol remained above 98%, which indicated that the leakage rate of the two drugs was within 2%.

[Test Example 3] Cytotoxicity of MCF-7 in combination with docetaxel and resveratrol.

Precisely weigh docetaxel and resveratrol, dissolve in a small amount of DMSO respectively, dilute docetaxel to 20.0 μg/mL concentration with DMEM culture medium; dilute resveratrol to 10.0, 20.0, 40.0, 80.0 μg/mL series concentration. Docetaxel and resveratrol were mixed in equal volumes to prepare a series of drug mixed solutions with concentration ratios of 2:1, 1:1, 1:2, and 1:4, respectively. (The concentration of the drug to be tested should be within the above concentration range.) The single-cell suspension of MCF-7 cells was inoculated in a 96-well cell culture plate (7×10 ³ cells per well) at 200 μL per well. Cultivate for 24 hours until the cells adhere to the wall, drain the culture solution, add different kinds of different concentrations of the test solution, and set up a zero-adjustment group and a control group (cell culture solution), and each group has 5 replicate wells. After culturing for 48 hours, add 20 μL of 5 mg·mL ^-1 MTT solution in PBS to each well, continue to cultivate for 4 hours, discard the culture medium, add 150 μL of dimethyl sulfoxide to each well, shake for about 10 minutes until the precipitate is completely dissolved, and measure each component , each ratio is used individually and jointly for the optical density value (OD value) at 490nm of MCF-7 cells for 48 hours.

When CDI<1, it shows a synergistic effect, when CDI<0.7, it shows an obvious synergistic effect; when CDI=1, it shows an additive effect, and when CDI>1, it shows an antagonistic effect. Figure 3a is the combined inhibitory effect of docetaxel and resveratrol on MCF-7, and Figure 3b is the combined effect index of the two drugs in different proportions. Alcohol concentration ratio in the range of 1:0.1-1:10 showed obvious synergistic effect.

[Test Example 4] Cytotoxic effect of combined docetaxel and resveratrol on MCF-7 stem cells

Culture of MCF-7 stem cells

MCF-7 cells were digested with trypsin, pipetted with DMEM-F12 medium, and placed in a culture flask at a density of 30,000 cells/mL for incubation. Under this culture condition, MCF-7 cells die gradually, while MCF-7 CSCs survive and non-adherently produce spherical cell clusters called mammospheres. Mammospheres can be collected by centrifugation at 1000rpm for 5 minutes. This culture method is called serum-free culture method.

Add 10,000 MCF-7 stem cells/well, and then add different concentrations of resveratrol solutions, and set up a zero-adjustment group and a control group (cell culture medium), and set 5 replicate wells in each well. After culturing for 48 hours, CellTiter96Aqueous reagent was added, and the OD value was measured with a microplate reader at 490 nm to calculate the cell survival rate.

It can be seen from Figure 4 that resveratrol has obvious killing effect on tumor stem cells, and the effect of inducing apoptosis is more obvious as the concentration increases. It shows that resveratrol can induce apoptosis of MCF-7 tumor cells.

[Test Example 5] The pharmacodynamic effect of docetaxel and resveratrol drug-loaded micelles on MCF-7 tumor-bearing mice

Detect the anticancer activity of 8-10 compositions in the embodiment

Culture MCF-7 cells stored in liquid nitrogen with DMEM medium containing 10% FBS, digest with trypsin before use, centrifuge at 1000rpm for 4min, take out the supernatant, and resuspend the cells in 0.9% normal saline , adjust the cell concentration to 5×10 ⁶ cells/200 μL, and then use a 1 mL syringe to inoculate the cell suspension into the right axilla of nude mice at a liquid volume of 200 μL per nude mouse. When the tumor grows to 50mm, start administration, and record the day ^of administration as day 0, nude mice are divided into 5 groups, on the 0th day, the 3rd day and the 6th day, respectively with 35mg/kg docetaxel , the dosage of 35mg/kg resveratrol, carry out by tail vein and give control group physiological saline, composition 8 (resveratrol micelle composition), composition 9 (docetaxel micelle composition) and combination Compound 10 (resveratrol/docetaxel polymer micelle composition).

Tumor volume was measured over time. Tumor volume was calculated according to Equation 1.

Formula 1

Tumor volume (TV)=(W ² ×L)/2 (where L is the long axis and W is the short axis)

To confirm the validity of the assay, use at least 4 nude mice per treatment and at least 4 tumors per group. At the starting point of treatment, the size of the tumor was: volume 50 mm ³ . If the animal died within 2 weeks of the last dose, it was considered a toxic death and was excluded from the evaluation. Groups in which more than one of the three animals died, or included groups that lost more than 15% of body weight without complete recovery, were considered to have no anticancer activity. The result is shown in Figure 5.

As can be seen from Figure 5, administration of the micellar composition of resveratrol (composition 8) has a certain inhibitory effect on cancer compared with the control group, but not as good as the micellar group of docetaxel (composition 9). Obviously (P<0.05). In particular, compared with the group treated with the mixed polymer micelle composition (composition 10) and the group treated with the paclitaxel-containing polymer micelle composition (composition 9) (P<0.05), the former showed more Good inhibitory activity on cancer cell growth.

Claims (10)

1. be used for the treatment of a polymer micelle composition for easy relapsed cancer, comprise Amphipathilic block polymer and active component, it is characterized in that, described active component is taxane and tumor stem cell apoptosis promoter resveratrol.

2. according to the polymer micelle composition being used for the treatment of easy relapsed cancer described in claim 1, it is characterized in that: described compositions comprises and is encapsulated in taxane in micelle and resveratrol simultaneously, or the mixture of encapsulating taxane polymer micelle and encapsulating resveratrol polymer micelle, or the compositions of the taxane be encapsulated in a micelle and resveratrol and encapsulating taxane polymer micelle and the mixture encapsulating resveratrol polymer micelle.

3. the polymer micelle composition being used for the treatment of easy relapsed cancer according to claim 1 or 2; it is characterized in that: described taxane is selected from the every mixture of paclitaxel, Docetaxel, 7-acetyl paclitaxel, t-acetyl paclitaxel, 10-deacetylate paclitaxel, 10-deacetylate-7-Epitaxol, 7-xyloside paclitaxel, 10-deacetylate-7-glutarylpaclitaxel, 7-N, N-dimethylglycinamidyl paclitaxel, 7-L-alanyl paclitaxel or more.

4. according to the polymer micelle composition that be used for the treatment of easy relapsed cancer of claim 1-3 described in any one, it is characterized in that: described amphipathic nature block polymer is the polymer comprising hydrophilic block and hydrophobic block, wherein, hydrophilic block is selected from one or more blocks of Polyethylene Glycol and methoxy poly (ethylene glycol); Hydrophobic block is selected from one or more blocks of the following: poly-newborn sour ﹑ gathers and hands over the acid of ester ﹑ polylactic acid second alcohol acid ﹑ PVOH to hand over ester ﹑ to gather flat Fructus Persicae acid ﹑ to gather own acid lactone ﹑ and gather and gather ammonia base acid ﹑ to dioxa Ketohexamethylene ﹑ and gather former sour ester ﹑ polyanhydride and above every copolymer.

5. the polymer micelle composition being used for the treatment of easy relapsed cancer according to claim 4, its feature is being: described hydrophilic block number-average molecular weight is 500-20,000 dalton, described hydrophobic block number-average molecular weight is 500-20,000 dalton, and the content of hydrophilic block shared by described block copolymer gross mass is 20-80wt%.

6. the polymer micelle composition being used for the treatment of easy relapsed cancer according to claim 1 and 2, is characterized in that: the mass ratio of taxane and resveratrol is 0.1-10:1.

7. the polymer micelle composition being used for the treatment of easy relapsed cancer according to claim 1 or 2, is characterized in that: it is 0.1-45wt% that taxane and resveratrol summation account for described micelle composition gross mass.

8. the polymer micelle composition being used for the treatment of easy relapsed cancer according to claim 1 and 2, is characterized in that, described micelle composition by weight percentage, comprising:

The taxane of 0.2-30wt%

The resveratrol of 0.2-20wt%

The amphipathic nature block polymer of 50-99.6wt%.

9., according to the preparation method being used for the treatment of the polymer micelle composition of easy relapsed cancer described in claim 1, it is characterized in that, comprise the following steps:

(1) taxane, resveratrol and amphipathic nature block polymer dissolve in organic solvent;

(2) organic solvent is mixed with water, be uniformly mixed rear lyophilizing;

Or

(1) taxane, resveratrol dissolve in organic solvent with amphipathic nature block polymer respectively;

(2) organic solvent containing taxane obtained and the organic solvent containing resveratrol are mixed with water respectively, be uniformly mixed rear lyophilizing.

10. the preparation method being used for the treatment of easy relapsed cancer polymer micelle composition according to claim 9, it is characterized in that, described organic solvent is selected from: at least one of methanol, ethanol, propanol, butanols, amylalcohol, hexanol, acetone, acetonitrile, dichloromethane, chloroform, methyl ethyl ketone or ethyl acetate.