Medicine for treating cervical cancer and preparation method thereof

Abstract

The present invention discloses a drug for treating cervical cancer and a preparation method thereof, wherein the drug comprises: polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearyl phosphatidylcholine and / or soybean lecithin, polypeptide NGKSAVLF and anti-tumor drug active ingredients. The present invention prepares a drug with cervical cancer targeting by designing a polypeptide sequence and combining it with a specific phospholipid, which is beneficial to reducing the clinical dosage of the drug and improving the therapeutic effect, and is beneficial to further reducing the use cost while avoiding the side effects of the drug.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and specifically relates to a medicine for treating cervical cancer and a preparation method thereof. Background Art

[0002] Cervical cancer is the most common gynecological malignancy, usually occurring at the junction of the squamous epithelial cells of the cervical vaginal part or transitional zone and the columnar epithelial cells of the endocervical canal. 99.8% of cervical cancers are caused by human papillomavirus (HPV) infection, and 95% of women have at least one chance of being infected with papillomavirus (HPV) in their lifetime. It is true that papillomavirus (HPV) is the culprit, but the human body will not sit idly by. Women have the ability to "detoxify" and will not let papillomavirus (HPV) do whatever it wants in the body. Generally, after being infected with papillomavirus (HPV), it will be excreted from the body within 8-12 months, thus maintaining a healthy state. The real reason is that women’s own resistance is low, which causes the human papillomavirus (HPV) to remain in the junction of the cervical squamous column for a long time. Generally, it will take more than 12 months to cause cervical intraepithelial neoplasia (CIN). It can be seen that persistent papillomavirus (HPV) infection is the real cause of cervical cancer, followed by the earliest cervical intraepithelial neoplasia, which is a low-grade lesion.

[0003] Traditional treatments for cervical cancer include surgery, radiotherapy, and chemotherapy. In recent years, traditional surgical resection treatments such as cryosurgery, laser therapy, and electrocoagulation have been added. Cryosurgery is only suitable for cases with small lesions and low CIN levels; recurrence is easy after laser treatment; electrocoagulation has disadvantages such as residual cancer lesions, secondary bleeding after surgery, and obvious pain; chemotherapy is not ideal and is mostly used for palliative treatment in the late stage; radiotherapy has satisfactory results but has a large reaction, especially the late reaction to the rectum, which causes patients to have continuous bloody stools, anemia, weakness, mental depression, and even loss of ability to work.

[0004] For example, Zhang Jinhui et al. explored the value of ultrasound contrast imaging in the staging of cervical cancer (Application of ultrasound contrast imaging in the staging of cervical cancer [J]. Journal of Clinical Ultrasound Medicine, 2013, 15(6):4.). They selected 30 patients with cervical cancer confirmed by pathology and undergoing conventional ultrasound and ultrasound contrast imaging, observed the relationship between cervical lesions and surrounding tissues, analyzed their contrast imaging data, and compared the morphology of the time-intensity curve and the differences in contrast imaging parameters. Results: 28 cases were diagnosed by ultrasound contrast imaging, with a diagnostic consistency rate of 93.33%; conventional ultrasound diagnosis 20 cases were diagnosed, and the diagnostic compliance rate was 66.67%. The difference between the two was statistically significant (P=0.01). The contrast characteristics were that when the drug was injected for 14.0s, the cervical cancer lesions appeared earlier than (16.0s) and higher than the myometrium. When the drug was injected for 26.7s, the drug in the lesion first disappeared and showed low enhancement, and the surrounding area showed slightly high enhancement, which could clearly show the infiltration range of cervical lesions. Compared with the surrounding normal uterine body tissue, the peak intensity of cervical cancer lesions was higher and the peak time was shortened, and the difference was statistically significant (P0.05). The research conclusion showed that the ultrasound contrast of cervical cancer had certain specificity and could assist clinical staging.

[0005] In the preliminary research process of the inventor of the present application, a contrast agent for ultrasound angiography of cervical cancer and its preparation method were proposed, and a Chinese invention patent application (application number CN202211153039.8) was submitted. The contrast agent includes: polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearyl phosphatidylcholine and / or soybean lecithin, polypeptide NGKSAVLF and perfluoroolefin gas. The prior application prepared an ultrasound contrast agent with cervical cancer targeting by designing a polypeptide sequence and combining it with a specific phospholipid, thereby improving the sensitivity and accuracy of ultrasound diagnosis of cervical cancer. Based on the prior research, the inventor of the present application used the polypeptide to develop drugs for the treatment of cervical cancer, thereby completing the present invention. The full text of the prior application is incorporated into a part of the specification of the present application, and the full text of the prior application is used as a part of the specification of the present application, and the applicant enjoys the right to modify and / or express opinions on the present application based on the full text of the prior application.

[0006] However, how to develop a drug that can enhance the treatment of cervical cancer remains a technical problem that needs to be solved urgently by technicians in this field. Summary of the invention

[0007] In view of the defects of the prior art, the present invention provides a drug that can enhance the treatment of cervical cancer. Specifically, in order to achieve the purpose of the present invention, the present invention intends to adopt the following technical solutions:

[0008] One aspect of the present invention relates to a drug for treating cervical cancer, characterized in that the drug comprises: polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearoyl phosphatidylcholine and / or soybean lecithin, polypeptide NGKSAVLF and anti-tumor drug active ingredients.

[0009] In a preferred embodiment of the present invention, the active ingredients of the anti-tumor drug are selected from: paclitaxel and / or fluorouracil.

[0010] In a preferred embodiment of the present invention, the weight ratio of polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearoyl phosphatidylcholine and / or soybean lecithin, polypeptide NGKSAVLF and active ingredient of anti-tumor drug is 2-4:3-6:2-4:0.2-0.5:1-2.

[0011] In a preferred embodiment of the present invention, the weight ratio of polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearoyl phosphatidylcholine and / or soybean lecithin, polypeptide NGKSAVLF and active ingredient of anti-tumor drug is 2-3:3-4:2-3:0.2-0.4:1-2.

[0012] In a preferred embodiment of the present invention, the drug is a sealed lyophilized powder injection or a suspension after reconstitution of the lyophilized powder injection.

[0013] In a preferred embodiment of the present invention, the average weight average molecular weight of the polyethylene glycol is 1500-3000.

[0014] The present invention also provides a method for preparing the above-mentioned medicine, comprising the following steps:

[0015] Polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearyl phosphatidylcholine and / or soybean lecithin are added to a mixed solvent of ethanol and water, homogenized at 20-40° C. and 80-120 MPa for 3-5 minutes, and then the polypeptide NGKSAVLF and the active ingredient of the anti-tumor drug are added. After homogenization for 0.5-2 minutes, the mixture is sterilized by filtering through a filter membrane to obtain an emulsion; the emulsion obtained in step (1) is packaged in vials and then freeze-dried in vacuum to obtain a lyophilized powder.

[0016] Beneficial effects of the present invention

[0017] The present invention prepares a drug with cervical cancer targeting by designing a polypeptide sequence and combining it with a specific phospholipid, which is beneficial to reducing the clinical dosage of the drug and improving the therapeutic effect, while helping to avoid drug side effects and further reducing the cost of use. DETAILED DESCRIPTION

[0018] In order to further understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0019] Unless otherwise specified, the reagents involved in the examples of the present invention are all commercially available products and can be purchased through commercial channels. The polypeptide sequence NGKSAVLF involved in the present invention was designed by the inventor and synthesized by Shanghai Bioengineering using solid phase synthesis.

[0020] Example 1: Preparation of a drug for the treatment of cervical cancer

[0021] 2 parts by weight of polyethylene glycol 2000, 2 parts by weight of distearyl phosphatidylcholine (DSPC), 3 parts by weight of dipalmitoyl phosphatidylglycerol sodium, 0.2 parts by weight of polypeptide NGKSAVLF, 100 parts by weight of 50 (v / v)% ethanol-water mixed solvent, and 1 part by weight of fluorouracil are prepared according to the following steps:

[0022] Add polyethylene glycol 2000, distearyl phosphatidylcholine and dipalmitoyl phosphatidylglycerol sodium to a mixed solvent of ethanol and water, homogenize at 30° C. and 100 MPa for 4 minutes, then add polypeptide and fluorouracil, homogenize for 1 minute, filter through a 220 nm filter membrane for sterilization, and obtain an emulsion; use vials to divide the emulsion obtained in step (1), and then vacuum freeze-dry to obtain a lyophilized powder to obtain the drug.

[0023] Embodiment 2:

[0024] The same as Example 1, except that paclitaxel is used instead of fluorouracil.

[0025] Embodiment 3:

[0026] The same as Example 1, except that the amount of polypeptide NGKSAVLF is adjusted to 0.1 parts by weight.

[0027] Comparative Example 1:

[0028] The same as Example 1, except that the polypeptide NGKSAVLF was not added.

[0029] Comparative Example 2:

[0030] The same as Example 1, except that 5 parts by weight of distearoyl phosphatidylcholine is used instead of the combination of distearoyl phosphatidylcholine and dipalmitoyl phosphatidylglycerol sodium.

[0031] Example 4: Zoological Experiment

[0032] (1) Cell culture: Human cervical cancer Hela cells were routinely cultured in a cell culture incubator at 37°C and 5% CO2 using DEME medium containing 10% calf serum.

[0033] (2) Construction of transplanted tumor animal model: 80 BALA / CA-nu nude mice, 5 weeks old and weighing 15-18 g, were selected and human cervical cancer Hela cells in logarithmic growth phase were injected subcutaneously into the back of nude mice. 1×10 6 Cells / piece.

[0034] (3) Dosage regimen: When the nude mouse tumor grows to a diameter of about 0.5 cm by naked eye observation, treatment is carried out. 21 mice with successful modeling were selected and randomly divided into a model group, a positive control group (commercially available fluorouracil injection, 15 fluorouracil per kilogram of body weight), Example 1-3 group and Comparative Example 1-2 group. Each group was given the corresponding drug (a suspension was made with 50 times the amount of normal saline, and 15 fluorouracil or paclitaxel was given per kilogram of body weight). The model group was injected with an equal amount of normal saline. The drug was administered once every three days for 3 consecutive times. The mice were killed by cervical dislocation 24 hours after the last administration, and the body weight was measured. The tumor mass was dissected and removed, and the tumor weight was measured. The tumor growth inhibition rate (%) of the drug administration group was calculated. Tumor inhibition rate (IR%) = (average tumor weight of the model group - average tumor weight of each treatment group) / average tumor weight of the model group) * 100%

[0035] (4) Experimental results: The experimental results are shown in Table 1. The experimental results show that the drug of the present invention has a relatively obvious effect of inhibiting tumor growth, and its inhibitory effect is significantly higher than that of the positive control group (there is a significant difference between the two groups), and its therapeutic effect is dose-dependent with the concentration of the polypeptide. This shows that the drug of the present invention can reduce the dosage of the active ingredient of the anti-tumor drug and improve the effect of treating cervical cancer.

[0036] Table 1. Effects of drugs on cervical cancer tumor weight

[0037] Group Tumor weight (mg) Tumor inhibition rate (%) Model Group 589±29 - Example 1 67±21 88.7 Example 2 78±15 86.8 Example 3 101±25 82.9 Comparative Example 1 269±19 54.4 Comparative Example 2 121±18 79.5 Positive Control 258±25 56.2

[0038] The above describes the preferred embodiments of the present invention, but it is not intended to limit the present invention. Those skilled in the art may make improvements and changes to the embodiments disclosed herein without departing from the scope and spirit of the present invention.

Claims

1. A drug for treating cervical cancer, characterized in that The drug comprises: polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearoyl phosphatidylcholine and / or soybean lecithin, polypeptide NGKSAVLF and active ingredients of anti-tumor drugs; The active ingredients of the anti-tumor drug are selected from: paclitaxel and / or fluorouracil; The weight ratio of the polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearoyl phosphatidylcholine and / or soybean lecithin, polypeptide NGKSAVLF and the active ingredient of the anti-tumor drug is 2-4:3-6:2-4:0.2-0.5:1-2; The average weight average molecular weight of the polyethylene glycol is 1500-3000.

2. The drug according to claim 1, wherein the weight ratio of polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearoyl phosphatidylcholine and / or soybean lecithin, polypeptide NGKSAVLF and the active ingredient of the anti-tumor drug is 2-3:3-4:2-3:0.2-0.4:1-2.

3. The medicine according to claim 1, which is a lyophilized powder injection.

4. The drug according to claim 1, which is a suspension after reconstitution of a lyophilized powder injection.

5. A method for preparing the drug according to any one of claims 1 to 4, comprising the following steps: Polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearyl phosphatidylcholine and / or soybean lecithin are added to a mixed solvent of ethanol and water, homogenized at 20-40°C and 80-120 MPa for 3-5 minutes, and then the polypeptide NGKSAVLF and the active ingredient of the anti-tumor drug are added. After homogenization for 0.5-2 minutes, the mixture is sterilized by filtering through a filter membrane to obtain an emulsion; the obtained emulsion is packaged in vials and then freeze-dried in vacuum to obtain a lyophilized powder.