CN115487321B - Contrast agent for ultrasound contrast of cervical cancer and preparation method thereof - Google Patents
Contrast agent for ultrasound contrast of cervical cancer and preparation method thereof Download PDFInfo
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- CN115487321B CN115487321B CN202211153039.8A CN202211153039A CN115487321B CN 115487321 B CN115487321 B CN 115487321B CN 202211153039 A CN202211153039 A CN 202211153039A CN 115487321 B CN115487321 B CN 115487321B
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- 206010008342 Cervix carcinoma Diseases 0.000 title claims abstract description 25
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
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- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical group CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 claims 1
- ATBOMIWRCZXYSZ-XZBBILGWSA-N [1-[2,3-dihydroxypropoxy(hydroxy)phosphoryl]oxy-3-hexadecanoyloxypropan-2-yl] (9e,12e)-octadeca-9,12-dienoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCC\C=C\C\C=C\CCCCC ATBOMIWRCZXYSZ-XZBBILGWSA-N 0.000 claims 1
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- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
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- 244000191761 Sida cordifolia Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
- A61K49/221—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by the targeting agent or modifying agent linked to the acoustically-active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
- A61K49/222—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by a special physical form, e.g. emulsions, liposomes
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Acoustics & Sound (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Radiology & Medical Imaging (AREA)
- Epidemiology (AREA)
- Physics & Mathematics (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a contrast agent for ultrasound contrast of cervical cancer and a preparation method thereof, wherein the contrast agent comprises the following components: polyethylene glycol, dipalmitoyl phosphatidylglycerol or salts thereof, distearyl phosphorylcholine and/or soybean phospholipids, polypeptide NGKSAVLF and perfluoroolefin gas. According to the invention, the polypeptide sequence is designed and combined with specific phospholipids to prepare the ultrasound contrast agent with cervical cancer targeting, so that the sensitivity and accuracy of ultrasound diagnosis of cervical cancer are improved.
Description
Technical Field
The invention belongs to the field of diagnostic reagents, and particularly relates to a contrast agent for ultrasound contrast of cervical cancer and a preparation method thereof.
Background
Cervical cancer is one of the common malignant tumors of gynaecology, and in recent years, the number of light patients is increased, and the accurate diagnosis of cervical cancer has important significance for the selection of the treatment mode and the treatment effect of the cervical cancer. Ultrasound is a noninvasive and low-cost examination method, and particularly, the rapid development of ultrasound radiography technology can display microcirculation changes in focuses, thereby providing more reliable imaging information for clinic.
For example, zhang Jinhui et al discuss the value of ultrasound imaging in cervical cancer staging (application of ultrasound imaging in cervical cancer staging [ J ]. Journal of clinical ultrasound medicine, 2013, 15 (6): 4.), 30 cervical cancer patients subjected to pathology verification and parallel conventional ultrasonic and ultrasonic radiography examination are selected, the relationship between cervical lesions and surrounding tissues is observed, radiography data of the cervical lesions are analyzed, and the differences of morphology and radiography parameters of time-intensity curves are compared; the contrast characteristic is that when contrast agent is injected for 14.0s, cervical cancer focus is earlier than (16.0 s) and higher than myometrium development, when the contrast agent is injected for 26.7s, the contrast agent in focus is firstly resolved, is low in enhancement, the periphery is slightly high in enhancement, and the infiltration range of cervical lesions can be clearly displayed. The research conclusion shows that the ultrasonic radiography of cervical cancer shows a certain specificity and can assist clinical staging.
Ultrasound contrast agents are a class of diagnostic agents capable of significantly enhancing the ultrasound detection signal, which generally comprise a microbubble structure, with strong scattering properties for ultrasound. In the above studies, the ultrasound contrast agent used was sonorovird. However, the sonorovird is a systemic contrast agent without directional selection, and has lower sensitivity and low specificity. In order to further enhance the targeting imaging enhancement effect of the ultrasonic contrast agent, the research of the targeting ultrasonic contrast agent is more and more, the existing targeting ultrasonic contrast agent often utilizes means such as antigen-antibody, biotin-avidin and the like to realize the targeting effect, and the method has the defects of complex preparation method, high requirements on storage and transportation conditions, high use cost and the like, thereby limiting the application of the targeting ultrasonic contrast agent.
For example, chinese patent CN113041365B discloses a targeted ultrasound contrast agent for enhancing carotid arteriosclerosis ultrasound diagnosis and a preparation method thereof, wherein the targeted ultrasound contrast agent for enhancing carotid arteriosclerosis ultrasound diagnosis is prepared from phospholipid and bio-inert gas containing hyaluronic acid, optionally mannose and derivatives thereof, carotid arteriosclerosis targeting of the ultrasound contrast agent is realized at low cost, and carotid arteriosclerosis ultrasound imaging effect is enhanced.
For another example, chinese patent CN111686263B discloses a targeted ultrasound contrast agent for enhancing lower limb arteriosclerosis ultrasound diagnosis and a preparation method thereof, wherein the targeted ultrasound contrast agent for enhancing lower limb arteriosclerosis ultrasound diagnosis is prepared from an ultrasound contrast agent of amino modified lipid membrane and fibrin targeted polypeptide, so that enrichment of the contrast agent on affected parts of lower limb arteriosclerosis is realized, the contrast duration is obviously prolonged, and the ultrasound imaging effect of lower limb arteriosclerosis is obviously enhanced.
However, how to develop a contrast agent capable of enhancing ultrasound contrast of cervical cancer is still a technical problem to be solved by those skilled in the art.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a contrast agent capable of enhancing ultrasound contrast of cervical cancer. Specifically, in order to achieve the purpose of the present invention, the present invention adopts the following technical scheme:
in one aspect, the invention relates to a contrast agent for ultrasound imaging of cervical cancer, characterized in that it comprises: polyethylene glycol, dipalmitoyl phosphatidylglycerol or salts thereof, distearyl phosphorylcholine and/or soybean phospholipids, polypeptide NGKSAVLF and perfluoroolefin gas.
In a preferred embodiment of the present invention, the perfluoroolefin gas is selected from the group consisting of: one or more combinations of perfluoropropane, perfluorobutane, perfluoropentane, perfluorohexane, perfluorooctane; more preferably, the perfluoroolefin gas is selected from the group consisting of: perfluoropropane and/or perfluorobutane.
In a preferred embodiment of the present invention, the weight ratio of polyethylene glycol, dipalmitoyl phosphatidylglycerol or salts thereof, distearyl phosphorylcholine and/or soybean phospholipids, polypeptide NGKSAVLF is 2-4:3-6:2-4:0.2-0.5.
In a preferred embodiment of the present invention, the weight ratio of polyethylene glycol, dipalmitoyl phosphatidylglycerol or salts thereof, distearyl phosphorylcholine and/or soybean phospholipids, polypeptide NGKSAVLF is 2-3:3-4:2-3:0.2-0.4.
In a preferred embodiment of the present invention, the contrast agent is a freeze-dried powder injection in sealed package.
In a preferred embodiment of the invention, the polyethylene glycol has an average weight average molecular weight of 1500-3000.
The invention also provides a preparation method of the ultrasonic contrast agent, which comprises the following steps:
(1) Adding polyethylene glycol, dipalmitoyl phosphatidylglycerol or its salt, distearyl phosphatidylcholine and/or soybean phospholipids into ethanol-water mixed solvent, homogenizing at 20-40deg.C under 80-120MPa for 3-5 min, adding polypeptide NGKSAVLF, homogenizing for 0.5-2 min, and filtering with filter membrane for sterilization to obtain emulsion;
(2) Sub-packaging the emulsion obtained in the step (1) by using a penicillin bottle, and then performing vacuum freeze-drying to obtain freeze-dried powder, slowly filling perfluoroolefin gas in a vacuum state, then slowly filling a mixed gas of the perfluoroolefin gas and nitrogen, and then sealing the penicillin bottle by using a rubber plug, wherein the volume ratio of the perfluoroolefin gas in the mixed gas is 40-60%;
(3) And (5) carrying out irradiation sterilization on the sealed freeze-dried powder sub-packaging container.
The invention has the beneficial effects that
According to the invention, the polypeptide sequence is designed, and the polypeptide sequence and the specific phospholipid are combined to prepare the ultrasound contrast agent with cervical cancer targeting, so that the sensitivity and accuracy of ultrasound diagnosis of cervical cancer are improved; on the other hand, the preparation method of the ultrasonic contrast agent prepared by the invention is simple and feasible, has low cost and is very beneficial to wide application of clinical diagnosis; in a further aspect, the ultrasound contrast agent prepared by the invention is beneficial to reducing the clinical dosage of the ultrasound contrast agent, and is beneficial to avoiding the side effect of the contrast agent and simultaneously is beneficial to further reducing the use cost.
Drawings
Fig. 1 and 2 are graphs showing the results of ultrasonic diagnosis using the contrast agent prepared in example 1.
Detailed Description
In order to further understand the present invention, a technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Unless otherwise specified, all reagents involved in the examples of the present invention are commercially available products and are commercially available. The polypeptide sequence NGKSAVLF related by the invention is designed by the inventor and is synthesized by Shanghai engineering by adopting a solid-phase synthesis method.
Example 1: targeted ultrasound contrast agent for ultrasound diagnosis of cervical cancer
2000 parts of polyethylene glycol, 2 parts of distearyl phosphorylcholine (DSPC), 3 parts of dipalmitoyl sodium phosphoryl glyceride, 0.2 part of polypeptide NGKSAVLF, 100 parts of 50 (v/v)% ethanol water mixed solvent and a proper amount of perfluoropropane, and is prepared according to the following steps:
(1) Adding polyethylene glycol 2000, distearyl phosphorylcholine and dipalmitoyl sodium phosphoryl glyceride into a mixed solvent of ethanol and water, homogenizing for 4 minutes at 30 ℃ and 100MPa, adding polypeptide, homogenizing for 1 minute, and filtering and sterilizing by a 220nm filter membrane to obtain emulsion;
(2) Sub-packaging the emulsion obtained in the step (1) by using a penicillin bottle, vacuum freeze-drying to obtain freeze-dried powder, slowly filling perfluoropropane to 55KPa under vacuum, slowly filling a mixed gas of perfluoropropane and nitrogen to 1 standard atmosphere, and sealing the penicillin bottle by using a rubber plug, wherein the volume ratio of perfluoropropane in the mixed gas is 50%;
(3) The sealed freeze-dried powder sub-packaging container is used 60 Co irradiation sterilization is carried out, and the targeted ultrasonic contrast agent for ultrasonic diagnosis of cervical cancer is obtained.
Example 2:
the same as in example 1, except that soybean phospholipid was used instead of distearylcholine.
Example 3:
the procedure is as in example 1, except that the amount of polypeptide NGKSAVLF is adjusted to 0.1 parts by weight.
Comparative example 1:
the same as in example 1, except that the polypeptide NGKSAVLF was not added.
Comparative example 2:
the same as in example 1, except that 5 parts by weight of distearylcholine was used instead of the combination of distearylcholine and sodium dipalmitoyl phosphatidyl glycerol.
Example 4: animal experiments
(1) Cell culture: DEME culture with 10% calf serum was performed at 37℃on 5% CO 2 The human cervical carcinoma Hela cells were routinely cultured in a cell incubator.
(2) Constructing an animal model of the transplanted tumor: 80 BALA/CA-nu nude mice are selected, the age of the mice is 5 weeks, the weight of the mice is 15-18g, logarithmic growth phase human cervical carcinoma Hela cells are taken to be injected subcutaneously on the back of the nude mice, and each injection is 1 multiplied by 10 6 Individual cells/individual.
(3) Ultrasound contrast examination: and when the tumor body of the nude mice is observed to be about 0.5cm in diameter by naked eyes, carrying out ultrasonic contrast examination on the nude mice with tumors. 21 mice with successful modeling were selected and randomly divided into a model group, a positive control group (sonolover), examples 1-3 groups and comparative examples 1-2 groups, each group corresponding to 0.1mL of ultrasound contrast agent (suspension prepared by 10 times of physiological saline), and the model group was injected with an equal amount of physiological saline. Using an ultrasonic diagnostic apparatus, the probe frequency is 10MHz, and the ultrasonic contrast peak intensity ratio of the tumor body parts of each group of rats is recorded, wherein:
peak intensity ratio = experimental group average peak intensity/model group average peak intensity.
(4) Experimental results: the experimental results are shown in table 1, and the experimental results show that the ultrasonic contrast agent provided by the invention has a relatively obvious ultrasonic enhancement effect, and the effect is higher than that of the positive control group of the sonorovird. In particular, example 1 of the present invention shows a more excellent cervical cancer targeting effect.
TABLE 1 influence of targeted ultrasound contrast Agents on ultrasound diagnosis of cervical cancer
Example 5: clinical trial
1 cervical cancer patient who is admitted to our hospital in 6 months 2022 is diagnosed by pathological result diagnosis. The patient had no history of heart disease, contrast agent and other allergies, informed consent for post-booked ultrasound contrast exam. The patient takes the supine position after holding urine, firstly, the patient is subjected to conventional ultrasonic examination of abdomen and gynecology, the position, the size, the shape, the echo characteristics and the like of the focus are recorded, and conventional ultrasonic diagnosis is made. The best section capable of clearly showing the focus is selected, the mode is switched to a contrast mode, 1.2ml of the contrast agent prepared in example 1 is rapidly infused through the elbow vein (10 times of normal saline is adopted to prepare suspension), a timer is started at the same time, 120S is continuously observed in real time, the process is stored in an instrument hard disk, and after the contrast is finished, the contrast image is replayed and analyzed to make ultrasonic contrast diagnosis. The experimental results are shown in fig. 1-2, and the experimental results show that the contrast agent provided by the invention can remarkably improve the contrast at the cervical.
The foregoing describes preferred embodiments of the present invention, but is not intended to limit the invention thereto. Modifications and variations to the embodiments disclosed herein may be made by those skilled in the art without departing from the scope and spirit of the invention.
Claims (4)
1. A contrast agent for ultrasound contrast of cervical cancer, characterized in that the contrast agent consists of polyethylene glycol, dipalmitoyl sodium phosphatidyl glycerol, distearyl phosphorylcholine and/or soybean lecithin, polypeptide NGKSAVLF and perfluoroolefin gas; the weight ratio of the polyethylene glycol to the sodium dipalmitoyl phosphatidyl glycerol to the distearyl phosphatidyl choline to the soybean lecithin to the polypeptide NGKSAVLF is 2-4:3-6:2-4:0.2-0.5; the perfluoroolefin gas is selected from the group consisting of: one or more combinations of perfluoropropane, perfluorobutane, perfluoropentane, perfluorohexane, perfluorooctane; the average weight average molecular weight of the polyethylene glycol is 1500-3000;
the ultrasonic contrast agent is prepared by a preparation method comprising the following steps:
(1) Adding polyethylene glycol, sodium dipalmitoyl phosphatidyl glycerol, distearyl phosphatidyl choline and/or soybean lecithin into a mixed solvent of ethanol and water, homogenizing for 3-5 minutes at 20-40 ℃ and 80-120MPa, adding polypeptide NGKSAVLF, homogenizing for 0.5-2 minutes, and filtering and sterilizing by a filter membrane to obtain emulsion;
(2) Sub-packaging the emulsion obtained in the step (1) by using a penicillin bottle, and then performing vacuum freeze-drying to obtain freeze-dried powder, slowly filling perfluoroolefin gas in a vacuum state, then slowly filling a mixed gas of the perfluoroolefin gas and nitrogen, and then sealing the penicillin bottle by using a rubber plug, wherein the volume ratio of the perfluoroolefin gas in the mixed gas is 40-60%;
(3) And (5) carrying out irradiation sterilization on the sealed freeze-dried powder sub-packaging container.
2. The contrast agent of claim 1, the perfluoroolefin gas being selected from the group consisting of: perfluoropropane and/or perfluorobutane.
3. The contrast agent according to claim 1, wherein the weight ratio of polyethylene glycol, sodium dipalmitoyl phosphatidyl glycerol, distearyl phosphorylcholine and/or soybean phospholipid, polypeptide NGKSAVLF is 2-3:3-4:2-3:0.2-0.4.
4. The contrast agent according to claim 1, which is a freeze-dried powder injection in sealed package.
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| CN105664188A (en) * | 2016-02-24 | 2016-06-15 | 袁哲 | Ultrasonic contrast agent of uterus oviduct tract and preparation method of ultrasonic contrast agent |
| CN107184994A (en) * | 2017-06-06 | 2017-09-22 | 山东省千佛山医院 | A kind of liver cancer Ultrasonic Diagnosis targeting agent and preparation method thereof |
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| CN105664188A (en) * | 2016-02-24 | 2016-06-15 | 袁哲 | Ultrasonic contrast agent of uterus oviduct tract and preparation method of ultrasonic contrast agent |
| CN107184994A (en) * | 2017-06-06 | 2017-09-22 | 山东省千佛山医院 | A kind of liver cancer Ultrasonic Diagnosis targeting agent and preparation method thereof |
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| Title |
|---|
| 郑荣琴等.超声造影新技术临床应用.广东科技出版社,2007,(第第1版版),第6-7页. * |
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