Ultrasonic imaging composite gel, preparation method therefor, and use thereof

Abstract

The present application relates to the technical field of pharmaceutical formulations and particularly to an ultrasonic imaging composite gel, a preparation method therefor, and use thereof. The present application provides an ultrasonic imaging composite gel comprising a gel matrix, an ultrasonic imaging material, and a pharmaceutically acceptable solvent. The concentration of the gel matrix in the ultrasonic imaging composite gel is 1-500 mg / mL, and the concentration of the ultrasonic imaging material in the ultrasonic imaging composite gel is 1-500 mg / mL. The gel matrix comprises one or more of a temperature-sensitive gel matrix, an injectable gel matrix, and an ion-responsive gel matrix, and the ultrasonic imaging material comprises an ultrasonic phase change imaging agent and a high-molecular-weight material wrapping the ultrasonic phase change imaging agent. The ultrasonic imaging material is an ultrasonic imaging microbubble structure. The ultrasonic imaging composite gel can enhance ultrasonic imaging during embryo transfer and promote the adhesion of embryos to the endometrium and the implantation of the embryos.

Description

An ultrasonic imaging composite gel, its preparation method and application

[0001] This application claims priority to Chinese Patent Application No. 202411755472.8, filed on December 2, 2024, entitled "An Ultrasonic Imaging Composite Gel and Its Preparation Method and Application", the contents of which are incorporated herein by reference. Technical Field

[0002] This application relates to the field of pharmaceutical formulation technology, and in particular to an ultrasonic imaging composite gel, its preparation method, and its application. Background Technology

[0003] Assisted reproductive technology (ART) is currently an effective treatment for human fertility-related problems, with in-vitro fertilization-embryo transfer (IVF-ET) being the most widely used. Embryo quality, endometrial receptivity, and synchronous development of the embryo and endometrium are the three key factors in embryo transfer. However, in current clinical embryo transfer procedures, problems such as weak adhesion between free embryos and the endometrium leading to ectopic pregnancies and poor endometrial receptivity at the embryo implantation site significantly limit the clinical pregnancy rate of ART. Furthermore, the current embryo transfer process requires abdominal ultrasound guidance, and uterine ultrasound visualization relies primarily on bladder fullness, resulting in imperfect visualization methods, poor patient compliance, and a less than ideal experience.

[0004] Currently, there are very few products available clinically for assisting embryo implantation. EmbryoGlue, an embryo culture medium produced by the Swedish reagent company Vitrolif, primarily consists of high-concentration hyaluronic acid and recombinant human albumin, designed to aid embryo implantation. However, this embryo culture medium has weak embryo adhesion, failing to stably attach the embryo to a specific endometrial site, potentially posing a high risk of ectopic pregnancy. Furthermore, the medium's fluidity means that nutrient loss may prevent complete supply to the embryonic tissue. In addition, existing patents primarily focus on improving the ultrasound imaging material of the embryo transfer catheter, such as patent CN213606766U, which designs the support core of the embryo transfer catheter as an ultrasound-enhancing material to more clearly observe the catheter placement process. However, due to the lack of ultrasound enhancement for the embryo and surrounding tissues, it may be impossible to accurately observe the process of the embryo being injected into the uterus through the catheter and to monitor implantation. Therefore, developing a formulation that can provide a nutrient reservoir for early embryos, enhance ultrasound imaging during embryo transfer, and promote embryo adhesion and implantation to the endometrium is essential for improving the clinical pregnancy rate of assisted reproductive technology. Summary of the Invention

[0005] In view of this, the purpose of this application is to provide an ultrasound imaging composite gel, its preparation method, and its application. The ultrasound imaging composite gel can provide a nutrient reservoir for early embryos, enhance ultrasound imaging during embryo transfer, and promote embryo adhesion to the endometrium and implantation.

[0006] To achieve the above-mentioned objectives, this application provides the following technical solution:

[0007] This application provides an ultrasonic imaging composite gel, comprising a gel matrix, an ultrasonic imaging material, and a pharmaceutically acceptable solvent;

[0008] The concentration of the gel matrix in the ultrasonic imaging composite gel is 1–500 mg / mL;

[0009] The concentration of the ultrasonic imaging material in the ultrasonic imaging composite gel is 1–500 mg / mL;

[0010] The gel matrix includes one or more of temperature-sensitive gel matrices, injectable gel matrices, and ion-responsive gel matrices;

[0011] The ultrasonic imaging material includes an ultrasonic phase change developer and a polymeric material encapsulating the ultrasonic phase change developer, and the ultrasonic imaging material has an ultrasonic imaging microbubble structure.

[0012] Preferably, the temperature-sensitive gel matrix includes poloxamer 407, poloxamer 188, polyethylene glycol monomethyl ether-polylactic acid glycolic acid copolymer, polyethylene glycol monomethyl ether-polylactic acid glycolic acid-polylysine block copolymer, polyethylene glycol-polyL-aspartic acid derivative block copolymer, polyethylene glycol-polylactic acid-polyethylene glycol block copolymer, polylactic acid-polyethylene glycol-polylactic acid block copolymer, polyethylene glycol-polyDL-lactic acid block copolymer, polyglycolic acid-lactide-polyethylene glycol-polyglycolic acid block copolymer, polyethylene glycol-polyglycolic acid-lactide-polyethylene glycol block copolymer, polycaprolactone-polyethylene glycol-polyglycolic acid block copolymer, and polyglycolic acid-polyglycolic acid-polylysine block copolymer. One or more of the following: caprolactone block copolymer, polyethylene oxide, polydimethylsiloxane, methacrylate, polyacrylic acid, poly-N-isopropylacrylamide, polymethyl methacrylate-co-ethyl acrylate, chitosan, chitosan lactate, chitosan quaternary ammonium salt, chitosan hydrochloride, chitosan nitrate, chitosan sulfate, chitosan acetate, hydroxypropyl chitosan, carboxymethyl chitosan, cellulose, sodium carboxymethyl cellulose, carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, ethyl cellulose, carboxymethyl hydroxyethyl cellulose, carboxymethyl hydroxypropyl cellulose, and carboxymethyl ethyl cellulose.

[0013] Preferably, the injectable gel matrix includes one or more of carbomer, polycarbofil, gelatin, pectin, carrageenan, gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulose polysaccharides, ethylene polymers, acrylic resins, polyvinyl alcohol, and polycarboxylate.

[0014] Preferably, the ion-responsive gel matrix comprises one or more of sodium alginate, potassium alginate, calcium alginate, lithium alginate, sodium oxidized alginate, potassium oxidized alginate, calcium oxidized alginate, lithium oxidized alginate, calcium carbonate, calcium oxide, calcium phosphate, heparin calcium, calcium acetate, calcium gluconate, calcium lactate, calcium citrate, and calcium chloride.

[0015] Preferably, the mass ratio of the polymer material to the ultrasonic phase change imaging agent is 1:(1-10).

[0016] Preferably, the polymeric material includes polylactic acid-glycolic acid copolymer, polyethylene glycol monomethyl ether-polylactic acid-glycolic acid copolymer, polyethylene glycol monomethyl ether-polylactic acid-polylysine block copolymer, polyethylene glycol-polyL-aspartic acid derivative block copolymer, polyethylene glycol-polylactic acid-polyethylene glycol block copolymer, polylactic acid-polyethylene glycol-polylactic acid block copolymer, polyethylene glycol-polyDL-lactic acid block copolymer, and polyglycolic acid-lactide-polyethylene glycol-polyglycolic acid-lactide block copolymer. Copolymer, polyethylene glycol-polyglycolic acid lactide-polyethylene glycol block copolymer, polycaprolactone-polyethylene glycol-polycaprolactone block copolymer, lecithin, soybean lecithin, hydrogenated soybean lecithin, dipalmitoyl phosphatidylcholine, distearyl phosphatidylcholine, dimyristoyl phosphatidylcholine, dioleoyl lecithin, dipalmitoyl phosphatidylethanolamine, distearyl phosphatidylethanolamine-polyethylene glycol, dipalmitoyl phosphatidylglycerol, and distearyl phosphatidylglycerol.

[0017] Preferably, the ultrasonic phase change imaging agent includes one or more of perfluoromethane, perfluoroethane, perfluoropropane, perfluorobutane, perfluoropentane, perfluorocyclobutane, perfluorohexane, perfluorobromooctane, perfluorocrown ether, sulfur hexafluoride, sodium bicarbonate, ammonium bicarbonate, nitric oxide, and hydrogen.

[0018] Preferably, the pharmaceutically acceptable solvent includes embryo culture medium.

[0019] This application also provides a method for preparing the ultrasonic imaging composite gel described in the above technical solution, comprising the following steps:

[0020] A polymer material, an ultrasonic phase change imaging agent, and a pharmaceutically acceptable solvent are first mixed to obtain an ultrasonic imaging microbubble suspension.

[0021] The ultrasonic imaging microbubble suspension and the second gel matrix are mixed to obtain the ultrasonic imaging composite gel.

[0022] Preferably, the first mixing includes mixing the polymer material and the ultrasonic phase change imaging agent, followed by emulsification, removal of the organic phase and drying, and then mixing with the pharmaceutically acceptable solvent.

[0023] This application also provides the application of the ultrasound imaging composite gel described in the above technical solution or the ultrasound imaging composite gel prepared by the preparation method described in the above technical solution in the preparation of assisted embryo transfer and implantation preparations.

[0024] This application provides an ultrasound imaging composite gel, comprising a gel matrix, an ultrasound imaging material, and a pharmaceutically acceptable solvent; the concentration of the gel matrix in the ultrasound imaging composite gel is 1–500 mg / mL; the concentration of the ultrasound imaging material in the ultrasound imaging composite gel is 1–500 mg / mL; the gel matrix includes one or more of temperature-sensitive gel matrices, injectable gel matrices, and ion-responsive gel matrices; the ultrasound imaging material includes an ultrasound phase change imaging agent and a polymeric material encapsulating the ultrasound phase change imaging agent, and the ultrasound imaging material has an ultrasound imaging microbubble structure. The imaging microbubbles in the above-mentioned ultrasound imaging composite gel enhance ultrasound imaging function under ultrasound triggering, facilitating precise observation of the embryo implantation process. Furthermore, compared to existing embryo culture media, which are fluid liquids and cannot stably attach the embryo to a specific endometrial site, embryo migration may pose a higher risk of ectopic pregnancy. The ultrasound imaging composite gel adheres to the endometrium in a semi-solid gel state within the uterine cavity, to a certain extent restricting embryo movement within the uterine cavity, increasing the chance of embryo implantation into the endometrium, thereby facilitating embryo implantation. Therefore, the imaging microbubbles in the aforementioned ultrasound imaging composite gel can facilitate precise observation of the embryo implantation process and effectively adhere the embryo to a specific endometrial site. Attached Figure Description

[0025] Figure 1 is an ultrasonic imaging diagram of the ultrasonically developed microbubble suspension described in Examples 1 and 25-27;

[0026] Figure 2 shows the gelation of the ultrasonic imaging composite gel solutions described in Examples 1-3 in a constant temperature chamber at 37°C.

[0027] Figure 3 shows the injectability of the ultrasonic imaging composite gel described in Examples 4-6;

[0028] Figure 4 shows the imaging effect of local tissues after intramuscular injection of the ultrasound imaging composite gel described in Examples 2, 5, 7, 11, 15, 16, 20 and 24;

[0029] Figure 5 shows the birthing of pups in the control group, Example 2, Example 5, Example 7, Example 11, Example 15, Example 16, Example 20 and Example 24;

[0030] Figure 6 is a quantitative statistical chart of the number of offspring in the control group, Example 2, Example 5, Example 7, Example 11, Example 15, Example 16, Example 20 and Example 24. Detailed Implementation

[0031] This application provides an ultrasonic imaging composite gel, comprising, by weight parts, a gel matrix, an ultrasonic imaging material, and a pharmaceutically acceptable solvent;

[0032] The concentration of the gel matrix in the ultrasonic imaging composite gel is 1–500 mg / mL;

[0033] The concentration of the ultrasonic imaging material in the ultrasonic imaging composite gel is 1–500 mg / mL;

[0034] The gel matrix includes one or more of temperature-sensitive gel matrices, injectable gel matrices, and ion-responsive gel matrices;

[0035] The ultrasonic imaging material includes an ultrasonic phase change developer and a polymeric material encapsulating the ultrasonic phase change developer, and the ultrasonic imaging material has an ultrasonic imaging microbubble structure.

[0036] Unless otherwise specified, all raw materials used in this application are commercially available products well known to those skilled in the art.

[0037] In this application, the ultrasonic imaging composite gel is preferably in the form of a solution or a semi-solid gel.

[0038] In this application, the concentration of the gel matrix in the ultrasonic imaging composite gel is 1-500 mg / mL, preferably 5-300 mg / mL, and more preferably 5-200 mg / mL.

[0039] In this application, the gel matrix includes one or more of temperature-sensitive gel matrices, injectable gel matrices, and ion-responsive gel matrices; the temperature-sensitive gel matrix includes poloxamer 407 (F127), poloxamer 188 (F68), polyethylene glycol monomethyl ether-polylactic acid glycolic acid copolymer (mPEG-PLGA), polyethylene glycol monomethyl ether-polylactic acid glycolic acid-polylysine block copolymer (mPEG-PLGA-PLL), polyethylene glycol-polyL-aspartic acid derivative block copolymer (PEG-Pasp), polyethylene glycol-polylactic acid-polyethylene glycol block copolymer (PEG-PLA-PEG), polylactic acid-polyethylene glycol-polylactic acid block copolymer (PLA-PEG-PLA), polyethylene glycol-polyDL-lactic acid block copolymer (mPEG-PDLLA), polyglycolic acid-lactide-polyethylene glycol-polyglycolic acid-lactide block copolymer (PLGA-PEG-PLGA), polyethylene glycol- The following are included in the following formulations: poly(glycolic acid-lactide-polyethylene glycol) block copolymer (PEG-PLGA-PEG), polycaprolactone-polyethylene glycol-polycaprolactone block copolymer (PCL-PEG-PCL), polyethylene oxide (PEO), polydimethylsiloxane (PDMS), methacrylate, polyacrylic acid (PAA), poly(N-isopropylacrylamide), poly(methyl methacrylate-co-ethyl acrylate), chitosan, chitosan lactate, chitosan quaternary ammonium salt, chitosan hydrochloride, chitosan nitrate, chitosan sulfate, chitosan acetate, hydroxypropyl chitosan, carboxymethyl chitosan, cellulose, sodium carboxymethyl cellulose, carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, ethyl cellulose, carboxymethyl hydroxyethyl cellulose, carboxymethyl hydroxypropyl cellulose, and carboxymethyl ethyl cellulose, more preferably including poloxamer 407 (F127), poloxamer 188 (F68), carboxymethyl chitosan, or carboxymethyl cellulose. The injectable gel matrix preferably includes one or more of carbomer, polycarboferrite, gelatin, pectin, carrageenan, gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulose polysaccharides, ethylene polymers, acrylic resins, polyvinyl alcohol, and polycarboxylate, more preferably carbomer; the ion-responsive gel matrix preferably includes one or more of sodium alginate, potassium alginate, calcium alginate, lithium alginate, sodium alginate oxide, potassium alginate oxide, calcium alginate oxide, lithium alginate oxide, calcium carbonate, calcium oxide, calcium phosphate, heparin calcium, calcium acetate, calcium gluconate, calcium lactate, calcium citrate, and calcium chloride, more preferably sodium alginate, calcium alginate, calcium alginate oxide, or heparin calcium. In this application, when the gel matrix is ​​two or more of the above-mentioned specific selections, this application does not have any special limitation on the ratio of the above-mentioned specific substances, and they can be mixed in any ratio.

[0040] In this application, the gel matrix can gel within the uterus, and the gel formation promotes intrauterine adhesion.

[0041] In this application, the concentration of the ultrasonic imaging material in the ultrasonic imaging composite gel is 1-500 mg / mL, preferably 5-300 mg / mL, and more preferably 5-200 mg / mL.

[0042] In this application, the ultrasonic imaging material comprises an ultrasonic phase change developer and a polymeric material encapsulating the ultrasonic phase change developer, wherein the ultrasonic imaging material has an ultrasonic imaging microbubble structure. In this application, the mass ratio of the polymeric material to the ultrasonic phase change developer is preferably 1:(1-10), more preferably 1:(5-10), and most preferably 1:10.

[0043] In this application, the polymeric material preferably includes polylactic-co-glycolic acid copolymer (PLGA), polyethylene glycol monomethyl ether-polylactic-co-glycolic acid copolymer (mPEG-PLGA), polyethylene glycol monomethyl ether-polylactic-co-glycolic acid-polylysine block copolymer (mPEG-PLGA-PLL), polyethylene glycol-poly-L-aspartic acid derivative block copolymer (PEG-Pasp), polyethylene glycol-polylactic acid-polyethylene glycol block copolymer (PEG-PLA-PEG), polylactic acid-polyethylene glycol-polylactic acid block copolymer (PLA-PEG-PLA), polyethylene glycol-polyDL-lactic acid block copolymer (mPEG-PDLLA), polyglycolic acid-lactide-polyethylene glycol-polyglycolic acid block copolymer (PLGA-PEG-PLGA), and polyethylene glycol-polyglycolic acid-lactide-polyethylene glycol block copolymer. The polymeric material comprises one or more of the following: polyethylene glycol monomethyl ether-poly(lactic acid glycol)-poly(lycine) block copolymer (PCL-PEG-PCL), lecithin, soybean lecithin, hydrogenated soybean lecithin, dipalmitoylphosphatidylcholine, distearylphosphatidylcholine, dimyristoylphosphatidylcholine, dioleoyllecithin, dipalmitoylphosphatidylethanolamine, distearylphosphatidylethanolamine, distearylphosphatidylethanolamine-polyethylene glycol, dipalmitoylphosphatidylglycerol, and distearylphosphatidylglycerol; more preferably, it includes polyethylene glycol monomethyl ether-poly(lactic acid glycol)-poly(lycine)-poly(lycine)-poly(lactic acid glycol) block copolymer (mPEG-PLGA-PLL) or poly(lactic acid glycol) copolymer (PLGA). When the polymeric material comprises two or more of the above-mentioned specific selections, this application does not impose any special limitation on the ratio of the above-mentioned specific substances, and they can be mixed in any ratio.

[0044] In this application, the ultrasonic phase change imaging agent preferably includes one or more of perfluoromethane, perfluoroethane, perfluoropropane, perfluorobutane, perfluoropentane, perfluorocyclobutane, perfluorohexane, perfluorobromooctane, perfluorocrown ether, sulfur hexafluoride, sodium bicarbonate, ammonium bicarbonate, nitric oxide, and hydrogen; more preferably, it includes perfluorohexane or sodium bicarbonate; when the ultrasonic phase change imaging agent is two or more of the above-mentioned specific selections, this application does not have any special limitation on the ratio of the above-mentioned specific substances, and they can be mixed in any ratio.

[0045] In this application, the ultrasound imaging material is used for ultrasound imaging of embryo transfer.

[0046] In this application, the pharmaceutically acceptable solvent includes embryo culture medium. Preferably, the pharmaceutically acceptable solvent also includes one or more of water for injection, physiological saline, phosphate buffered saline solution, glucose solution, and enzyme-free water, more preferably water for injection or physiological saline. When the pharmaceutically acceptable solvent is two or more of the above-mentioned specific selections, this application does not have any special limitation on the ratio of the above-mentioned substances; they can be mixed in any ratio. In this application, the pharmaceutically acceptable solvent serves to disperse the above-mentioned gel matrix.

[0047] In this application, existing fluidized embryo culture media cannot form an in-situ nutrient reservoir, and nutrient loss is detrimental to pre-implantation nutrient supply. This application utilizes embryo culture media as a gel solvent, which, along with gel formation within the uterine cavity, continuously provides early nutrients to the embryo, thereby promoting implantation. Therefore, the nutrients in the gel (embryo culture media) can provide an effective nutrient reservoir for early embryo implantation.

[0048] This application also provides a method for preparing the ultrasonic imaging composite gel described in the above technical solution, comprising the following steps:

[0049] A polymer material, an ultrasonic phase change imaging agent, and a pharmaceutically acceptable solvent are first mixed to obtain an ultrasonic imaging microbubble suspension.

[0050] The ultrasonic imaging microbubble suspension and the second gel matrix are mixed to obtain the ultrasonic imaging composite gel.

[0051] This application first mixes a polymer material, an ultrasonic phase change imaging agent, and a pharmaceutically acceptable solvent to obtain an ultrasonic imaging microbubble suspension.

[0052] In this application, the first mixing preferably includes mixing a polymeric material and an ultrasonic phase change imaging agent, followed by emulsification, removal of the organic phase, and drying, and then mixing with the pharmaceutically acceptable solvent.

[0053] In this application, the emulsification preferably involves mixing the polymer material and the ultrasonic phase change developer with dichloromethane, then adding an emulsifier and performing ultrasonic emulsification. This application does not impose any special limitations on the amount of dichloromethane, the type of emulsifier, the amount of emulsifier, or the ultrasonic conditions; any method well-known to those skilled in the art can be used. In an embodiment of this application, the emulsification process can be as follows: mixing 10 mg of polymer material and 20 mg of ultrasonic phase change developer with 500 μL of dichloromethane, adding 1.5 mL of a 1% (w / w) polyvinyl alcohol aqueous solution, and performing ultrasonic emulsification.

[0054] In this application, the process of removing the organic phase is preferably vacuum removal, and the temperature of the vacuum removal is preferably room temperature; this application does not impose any special limitations on the specific process of the vacuum removal, and any process well known to those skilled in the art can be used.

[0055] In this application, the drying method is preferably freeze drying. This application does not have any special limitations on the freeze drying process, and any process well known to those skilled in the art can be used.

[0056] After obtaining the ultrasonic imaging microbubble suspension, this application mixes the ultrasonic imaging microbubble suspension with a second gel matrix to obtain the ultrasonic imaging composite gel.

[0057] In this application, the second mixing is preferably carried out under stirring conditions. This application does not impose any special limitations on the stirring process, and any process well known to those skilled in the art can be used.

[0058] After the second mixing is completed, if the pH of the resulting mixture does not meet the requirements of neutrality or reach a pH value similar to that of the human body (7.35-7.45), it is preferable to adjust it with a pH adjuster. This application does not have any special limitations on the adjustment process, and any process known to those skilled in the art can be used.

[0059] This application also provides the application of the ultrasound imaging composite gel described in the above technical solution or the ultrasound imaging composite gel prepared by the preparation method described in the above technical solution in the preparation of assisted embryo transfer and implantation preparations.

[0060] In this application, the preferred application is to add an intact in vitro embryo into the ultrasound imaging composite gel, and then, under abdominal ultrasound guidance, implant the prepared ultrasound imaging composite gel containing the embryo into the uterus.

[0061] The ultrasound imaging microbubbles in the ultrasound imaging composite gel described in this application facilitate the timing and positioning of embryo transfer surgery under abdominal ultrasound guidance. Based on the semi-solid adhesiveness of the ultrasound imaging composite gel, the embryo can be effectively adhered to a specific endometrial site, reducing the occurrence of ectopic pregnancy and promoting embryo implantation.

[0062] The technical solutions of this application will be clearly and completely described below with reference to the embodiments therein. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this application without creative effort are within the scope of protection of this application.

[0063] Example 1

[0064] 10 mg of polyethylene glycol monomethyl ether-polylactic acid glycol-polylysine block copolymer (mPEG-PLGA-PLL) and 200 μL of perfluorohexane were dissolved in 500 μL of dichloromethane, and then 1.5 mL of 1% polyvinyl alcohol aqueous solution was added for ultrasonic emulsification. The organic phase was then removed under vacuum at room temperature, followed by lyophilization. The resulting ultrasonically imaging microbubbles were dissolved in 1 mL of water for injection to obtain an ultrasonically imaging microbubble suspension.

[0065] Add 150 mg of poloxamer 407 to the ultrasonic imaging microbubble suspension, stir thoroughly in an ice-water bath until it swells and is completely dissolved to obtain an ultrasonic imaging composite gel solution.

[0066] Example 2

[0067] Referring to Example 1, the difference is that the dosage of poloxamer 407 is replaced with 170 mg.

[0068] Example 3

[0069] Referring to Example 1, the difference is that the dosage of poloxamer 407 is replaced with 200 mg.

[0070] The ultrasonic imaging composite gel solutions prepared in Examples 1-3 were placed in a constant temperature chamber at 37°C, and their gelation was observed by inverting the vials. Figure 2 shows the gelation of the ultrasonic imaging composite gel solutions in Examples 1-3 in a constant temperature chamber at 37°C. As can be seen from Figure 2, the ultrasonic imaging composite gel solutions in Examples 1-3 can all form gels.

[0071] Example 4

[0072] An ultrasonically imaging microbubble suspension was prepared according to Example 1;

[0073] Add 5 mg of carbomer to the ultrasonic imaging microbubble suspension, stir thoroughly to swell, and then adjust the pH to neutral with a 1 mol / L sodium hydroxide aqueous solution to obtain the ultrasonic imaging composite gel.

[0074] Example 5

[0075] Referring to Example 4, the difference is that the dosage of carbomer is adjusted to 10 mg.

[0076] Example 6

[0077] Referring to Example 4, the difference is that the dosage of carbomer is adjusted to 20 mg.

[0078] The ultrasonic imaging composite gels described in Examples 4-6 were placed in a 20-gauge syringe, and their injectability was observed. Figure 3 shows the injectability of the ultrasonic imaging composite gels described in Examples 4-6. As can be seen from Figure 3, the ultrasonic imaging composite gels described in Examples 4-6 are all injectable.

[0079] Example 7

[0080] An ultrasonically imaging microbubble suspension was prepared according to Example 1;

[0081] Add 5 mg of sodium alginate to the ultrasonic imaging microbubble suspension, stir thoroughly to dissolve and swell, then add 5 mg of calcium alginate and stir until homogeneous to obtain the ultrasonic imaging composite gel.

[0082] Example 8

[0083] Refer to Example 7, except that the amount of calcium alginate is replaced with 10 mg.

[0084] Example 9

[0085] Referring to Example 7, the difference is that the amount of calcium alginate is replaced with 15 mg.

[0086] Example 10

[0087] Referring to Example 7, the difference is that the amount of sodium alginate is replaced with 10 mg.

[0088] Example 11

[0089] Referring to Example 7, the difference is that the amount of sodium alginate is replaced with 10 mg, and the amount of calcium alginate is replaced with 10 mg.

[0090] Example 12

[0091] Referring to Example 7, the difference is that the amount of sodium alginate is replaced with 10 mg and the amount of calcium alginate is replaced with 15 mg.

[0092] Example 13

[0093] Referring to Example 7, the difference is that the amount of sodium alginate is replaced with 15 mg.

[0094] Example 14

[0095] Referring to Example 7, the difference is that the amount of sodium alginate is replaced with 15 mg and the amount of calcium alginate is replaced with 10 mg.

[0096] Example 15

[0097] Referring to Example 7, the difference is that the amount of sodium alginate is replaced with 15 mg, and the amount of calcium alginate is replaced with 15 mg.

[0098] The ultrasonic imaging composite gels described in Examples 7-15 were observed for gelation using the inverted vial method. At room temperature, the vial was inverted at regular intervals to observe gelation. The gelation time was defined as the time during which no gel flowed after inversion. The test results are shown in Table 1.

[0099] Table 1. Gel-forming properties and gel-forming time of the ultrasonic imaging composite gels described in Examples 7-15.

[0100] As shown in Table 1, the ultrasonic imaging composite gels described in Examples 7 to 15 can all be gelled. Among them, Example 15 has the shortest gelling time. This indicates that the gelling time of the ultrasonic imaging composite gel decreases with the increase of sodium alginate and calcium alginate concentrations.

[0101] Example 16

[0102] An ultrasonically imaging microbubble suspension was prepared according to Example 1;

[0103] 10 mg of sodium alginate was added to the ultrasonic imaging microbubble suspension, and after thorough stirring and swelling, 500 AxaIU of heparin injection was added and stirred evenly to obtain the ultrasonic imaging composite gel.

[0104] Example 17

[0105] Referring to Example 16, the difference is that the dosage of heparin injection is adjusted to 750 XaIU.

[0106] Example 18

[0107] Referring to Example 16, the difference is that the dosage of heparin injection is adjusted to 1000 XaIU.

[0108] Example 19

[0109] Referring to Example 16, the difference is that the amount of sodium oxidized alginate is adjusted to 20 mg.

[0110] Example 20

[0111] Referring to Example 16, the difference is that the dosage of sodium alginate was adjusted to 20 mg and the dosage of heparin injection was adjusted to 750 Xa IU.

[0112] Example 21

[0113] Referring to Example 16, the difference is that the dosage of sodium oxidized alginate was adjusted to 20 mg, and the dosage of heparin injection was adjusted to 1000 Xa IU.

[0114] Example 22

[0115] Referring to Example 16, the difference is that the amount of sodium oxidized alginate is adjusted to 30 mg.

[0116] Example 23

[0117] Referring to Example 16, the difference is that the dosage of sodium alginate was adjusted to 30 mg and the dosage of heparin injection was adjusted to 750 Xa IU.

[0118] Example 24

[0119] Referring to Example 16, the difference is that the dosage of sodium alginate was adjusted to 30 mg and the dosage of heparin injection was adjusted to 1000 Xa IU.

[0120] The ultrasonic imaging composite gels described in Examples 16-24 were observed for gelation using the inverted vial method. At room temperature, the vial was inverted at regular intervals to observe gelation. The gelation time was defined as the time at which no gel flowed after inversion. The test results are shown in Table 2.

[0121] Table 2. Gel-forming properties and gel-forming time of the ultrasonic imaging composite gels described in Examples 16-24.

[0122] As shown in Table 2, the ultrasonic imaging composite gels described in Examples 16-23 can all be gelled. Among them, Example 28 has the shortest gelling time. This indicates that the gelling time of the ultrasonic imaging composite gel decreases with increasing concentrations of sodium oxidized alginate and heparin calcium.

[0123] Example 25

[0124] Referring to Example 1, the difference is that the polyethylene glycol monomethyl ether-polylactic acid glycolic acid-polylysine block copolymer is replaced with polylactic acid glycolic acid copolymer.

[0125] Example 26

[0126] Referring to Example 1, the difference is that 200 μL of perfluorohexane was replaced with 20 mg of ammonium bicarbonate.

[0127] Example 27

[0128] Refer to Example 25, except that 200 μL of perfluorohexane was replaced with 20 mg of ammonium bicarbonate.

[0129] The performance of the ultrasound-enhanced ultrasound imaging of the ultrasound-enhanced microbubble suspensions described in Examples 1 and 25-27 was tested in vitro: The ultrasound-enhanced microbubble suspensions described in Examples 1 and 25-27 were loaded into the fingertips of latex gloves, and an ultrasound coupling agent was applied to the surface of the latex gloves. In vitro ultrasound imaging was performed on an ultrasound imaging instrument. Figure 1 shows the ultrasound imaging of the ultrasound-enhanced microbubble suspensions described in Examples 1 and 25-27. As can be seen from Figure 1, the ultrasound-enhanced ultrasound imaging of the ultrasound-enhanced microbubble suspensions described in Examples 1 and 25-27 can all be performed in vitro, with the ultrasound-enhanced imaging performance of Example 1 being more superior.

[0130] Test case

[0131] The imaging effects of local tissues were observed after intramuscular injection of 200 μL of the ultrasound imaging composite gel described in Examples 2, 5, 7, 11, 15, 16, 20, and 24. Figure 4 shows the imaging effects of local tissues after intramuscular injection of the ultrasound imaging composite gel described in Examples 2, 5, 7, 11, 15, 16, 20, and 24. As can be seen from Figure 4, the ultrasound imaging composite gel described in this application can enhance ultrasound imaging features in vivo.

[0132] Mice in pseudopregnancy at 6-8 weeks of gestation were randomly divided into 9 groups of 3 mice each. The oviduct was removed through a dorsal opening, and an oocyte transfer needle was inserted into the ampulla through the ovarian end of the oviduct opening. The negative control group received 10 mouse two-cell embryos containing culture medium. The experimental groups received 10 mouse two-cell embryos treated with the ultrasound-enhancing composite gel described in Examples 2, 5, 7, 11, 15, 16, 20, and 24, respectively. The number of pups born was observed. Figure 5 shows the pup births of the control group, Examples 2, 5, 7, 11, 15, 16, 20, and 24. Figure 6 shows the quantitative statistics of the number of pups born in the control group, Examples 2, 5, 7, 11, 15, 16, 20, and 24. As shown in Figures 5 and 6, the ultrasound-enhancing composite gel described in this application promotes assisted embryo implantation, with Example 20 showing the highest birth rate.

[0133] The above description is merely a preferred embodiment of this application and does not constitute any limitation on this application. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of this application, and these improvements and modifications should also be considered within the scope of protection of this application.

Claims

1. An ultrasonic imaging composite gel, characterized in that, This includes the gel matrix, ultrasound imaging materials, and pharmaceutically acceptable solvents; The concentration of the gel matrix in the ultrasonic imaging composite gel is 1–500 mg / mL; The concentration of the ultrasonic imaging material in the ultrasonic imaging composite gel is 1–500 mg / mL; The gel matrix includes one or more of temperature-sensitive gel matrices, injectable gel matrices, and ion-responsive gel matrices; The ultrasonic imaging material includes an ultrasonic phase change developer and a polymeric material encapsulating the ultrasonic phase change developer, and the ultrasonic imaging material has an ultrasonic imaging microbubble structure.

2. The ultrasonic imaging composite gel as described in claim 1, characterized in that, The ultrasonic imaging composite gel is a solution or a semi-solid gel.

3. The ultrasonic imaging composite gel as described in claim 1, characterized in that, The temperature-sensitive gel matrix includes poloxamer 407, poloxamer 188, polyethylene glycol monomethyl ether-polylactic acid glycolic acid copolymer, polyethylene glycol monomethyl ether-polylactic acid glycolic acid-polylysine block copolymer, polyethylene glycol-polyL-aspartic acid derivative block copolymer, polyethylene glycol-polylactic acid-polyethylene glycol block copolymer, polylactic acid-polyethylene glycol-polylactic acid block copolymer, polyethylene glycol-polyDL-lactic acid block copolymer, poly(lactic acid-lactide)-polyethylene glycol-poly(lactic acid-lactide) block copolymer, polyethylene glycol-poly(lactic acid-lactide)-polyethylene glycol block copolymer, and polycaprolactone-polyethylene glycol-polycaprolactone. One or more of the following: ester block copolymers, polyethylene oxide, polydimethylsiloxane, methacrylate, polyacrylic acid, poly-N-isopropylacrylamide, polymethyl methacrylate-co-ethyl acrylate, chitosan, chitosan lactate, chitosan quaternary ammonium salt, chitosan hydrochloride, chitosan nitrate, chitosan sulfate, chitosan acetate, hydroxypropyl chitosan, carboxymethyl chitosan, cellulose, sodium carboxymethyl cellulose, carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, ethyl cellulose, carboxymethyl hydroxyethyl cellulose, carboxymethyl hydroxypropyl cellulose, and carboxymethyl ethyl cellulose.

4. The ultrasonic imaging composite gel as described in claim 3, characterized in that, The temperature-sensitive types include poloxamer 407, poloxamer 188, carboxymethyl chitosan, or carboxymethyl cellulose.

5. The ultrasonic imaging composite gel as described in claim 1, characterized in that, The injectable gel matrix includes one or more of carbomer, polycarbofil, gelatin, pectin, carrageenan, gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulose polysaccharides, ethylene polymers, acrylic resins, polyvinyl alcohol, and polycarboxylate.

6. The ultrasonic imaging composite gel as described in claim 5, characterized in that, The injectable gel matrix includes carbomer.

7. The ultrasonic imaging composite gel as described in claim 1, characterized in that, The ion-responsive gel matrix includes one or more of sodium alginate, potassium alginate, calcium alginate, lithium alginate, sodium oxidized alginate, potassium oxidized alginate, calcium oxidized alginate, lithium oxidized alginate, calcium carbonate, calcium oxide, calcium phosphate, heparin calcium, calcium acetate, calcium gluconate, calcium lactate, calcium citrate, and calcium chloride.

8. The ultrasonic imaging composite gel as described in claim 6, characterized in that, The ion-responsive gel matrix includes sodium alginate, calcium alginate, oxidized calcium alginate, or heparin calcium.

9. The ultrasonic imaging composite gel as described in claim 1, characterized in that, The mass ratio of the polymer material to the ultrasonic phase change imaging agent is 1:(1-10).

10. The ultrasonic imaging composite gel as described in claim 1 or 9, characterized in that, The polymeric materials include polylactic acid-glycolic acid copolymer, polyethylene glycol monomethyl ether-polylactic acid-glycolic acid copolymer, polyethylene glycol monomethyl ether-polylactic acid-polylysine block copolymer, polyethylene glycol-polyL-aspartic acid derivative block copolymer, polyethylene glycol-polylactic acid-polyethylene glycol block copolymer, polylactic acid-polyethylene glycol-polylactic acid block copolymer, polyethylene glycol-polyDL-lactic acid block copolymer, and poly(lactic acid-lactide)-polyethylene glycol-poly(lactic acid-lactide) block copolymer. The following are included in the list of: polyethylene glycol-polyglycolic acid-lactide-polyethylene glycol block copolymer, polycaprolactone-polyethylene glycol-polycaprolactone block copolymer, lecithin, soybean lecithin, hydrogenated soybean lecithin, dipalmitoylphosphatidylcholine, distearylphosphatidylcholine, dimyristoylphosphatidylcholine, dioleoyllecithin, dipalmitoylphosphatidylethanolamine, distearylphosphatidylethanolamine-polyethylene glycol, dipalmitoylphosphatidylglycerol, and distearylphosphatidylglycerol.

11. The ultrasonic imaging composite gel as described in claim 10, characterized in that, The polymeric material includes polyethylene glycol monomethyl ether-polylactic acid glycolic acid-polylysine block copolymer or polylactic acid glycolic acid copolymer.

12. The ultrasonic imaging composite gel as described in claim 1 or 9, characterized in that, The ultrasonic phase change imaging agent includes one or more of perfluoromethane, perfluoroethane, perfluoropropane, perfluorobutane, perfluoropentane, perfluorocyclobutane, perfluorohexane, perfluorobromooctane, perfluorocrown ether, sulfur hexafluoride, sodium bicarbonate, ammonium bicarbonate, nitric oxide, and hydrogen.

13. The ultrasonic imaging composite gel as described in claim 12, characterized in that, The ultrasonic phase change imaging agent includes perfluorohexane or sodium bicarbonate.

14. The ultrasonic imaging composite gel as described in claim 1, characterized in that, Pharmaceutically acceptable solvents include embryo culture media.

15. The ultrasonic imaging composite gel as described in claim 14, characterized in that, Pharmaceutically acceptable solvents also include one or more of water for injection, physiological saline, phosphate buffered saline, glucose solution, and enzyme-free water.

16. A method for preparing the ultrasonic imaging composite gel according to any one of claims 1 to 15, characterized in that, Includes the following steps: A polymer material, an ultrasonic phase change imaging agent, and a pharmaceutically acceptable solvent are first mixed to obtain an ultrasonic imaging microbubble suspension. The ultrasonic imaging microbubble suspension and the second gel matrix are mixed to obtain the ultrasonic imaging composite gel.

17. The use of the ultrasound imaging composite gel according to any one of claims 1 to 15 or the ultrasound imaging composite gel prepared by the preparation method according to claim 16 in the preparation of assisted embryo transfer and implantation preparations.

18. A method for assisted embryo transfer and implantation, characterized in that, Includes the following steps: In vitro intact embryos are added to ultrasound-enhanced composite gels, and under abdominal ultrasound guidance, the prepared ultrasound-enhanced composite gel containing the embryos is implanted into the uterus. The ultrasonic imaging composite gel is the ultrasonic imaging composite gel according to any one of claims 1 to 15 or the ultrasonic imaging composite gel prepared by the preparation method according to claim 16.

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