An ultrasonic imaging composite gel and its preparation method and application

Abstract

The present application relates to the technical field of pharmaceutical preparation, in particular to an ultrasonic imaging composite gel, a preparation method and application thereof. The present application provides an ultrasonic imaging composite gel, which comprises a gel matrix, an ultrasonic imaging material and a pharmaceutically acceptable solvent according to mass fraction; 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 comprises one or more of a temperature-sensitive gel matrix, an injectable gel matrix and an ion-responsive gel matrix; the ultrasonic imaging material comprises an ultrasonic phase transition imaging agent and a polymer material wrapping the ultrasonic phase transition imaging agent, and the ultrasonic imaging material is an ultrasonic imaging microbubble structure. The ultrasonic imaging composite gel can enhance ultrasonic imaging in the process of embryo transplantation and promote the adhesion and implantation of embryos and endometrium.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical preparations, in particular to an ultrasonic imaging composite gel and a preparation method and application thereof. BACKGROUND

[0002] Assisted reproductive technology (ART) is an effective treatment method for solving human fertility-related problems at present. In particular, in vitro fertilization-embryo transfer technology is most widely used. Embryo quality, endometrial receptivity, and synchronous development of embryo and endometrium are the three key factors for embryo transfer. However, in the current clinical embryo transfer process, ectopic pregnancy caused by weak adhesion of free embryos to the endometrium and poor endometrial receptivity at the embryo implantation site greatly limit the clinical pregnancy rate of ART, causing great economic pressure and physical and mental suffering to patients. At the same time, the current embryo transfer process needs to be completed under the guidance of abdominal B-ultrasound, and the ultrasonic imaging of the uterus mainly relies on the filling of urine in the bladder, which has problems such as imperfect imaging method, poor patient compliance and experience, etc.

[0003] At present, there are few products for assisting embryo implantation in clinical practice. EmbryoGlu is a kind of embryo culture medium produced by Vitrolif, a Swedish reagent manufacturer, which mainly contains high-concentration hyaluronic acid and recombinant human albumin, aiming to help embryo implantation. However, the embryo culture medium has weak adhesion to the embryo, and cannot stably attach the embryo to a specific endometrial site, which may still have a high risk of ectopic pregnancy. At the same time, the embryo culture medium has a certain flowability, and the loss of nutrients may not completely supply the embryo tissue. In addition, the ultrasonic imaging method of embryo transfer in the existing patents mainly improves the ultrasonic imaging material of the embryo transfer catheter, such as the patent CN213606766U, which designs the support heart of the embryo transfer tube as an ultrasonic limiting enhancement material, so as to more clearly observe the embryo transfer catheter placement process. However, due to the lack of ultrasonic enhancement imaging of the embryo and its surrounding tissue, the embryo injection process into the uterus and the observation of embryo implantation may not be accurately observed. Therefore, it is necessary to develop a preparation that can provide a nutrient reservoir for early embryos, enhance ultrasonic imaging during embryo transfer, and promote embryo adhesion and implantation to the endometrium, in order to improve the clinical pregnancy rate of assisted reproductive technology. SUMMARY

[0004] Therefore, the purpose of the present application is to provide an ultrasonic imaging composite gel and a preparation method and application thereof. The ultrasonic imaging composite gel can provide a nutrient reservoir for early embryos, enhance ultrasonic imaging during embryo transfer, and promote embryo adhesion and implantation to the endometrium.

[0005] In order to achieve the above-mentioned purpose of the application, the present application provides the following technical solutions:

[0006] The present application provides an ultrasonic imaging composite gel, which comprises a gel matrix, an ultrasonic imaging material and a pharmaceutically acceptable solvent according to mass fraction;

[0007] The concentration of the gel matrix in the ultrasonic imaging composite gel is 1-500 mg / mL;

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

[0009] The gel matrix comprises one or more of a temperature-sensitive gel matrix, an injectable gel matrix and an ion-responsive gel matrix.

[0010] The ultrasonic imaging material comprises an ultrasonic phase transition imaging agent and a high molecular material wrapping the ultrasonic phase transition imaging agent, and the ultrasonic imaging material is an ultrasonic imaging microbubble structure.

[0011] Preferably, the temperature-sensitive gel matrix comprises one or more of poloxamer 407, poloxamer 188, polyethylene glycol monomethyl ether-poly (lactic acid-co-glycolic acid), polyethylene glycol monomethyl ether-poly (lactic acid-co-glycolic acid)-polylysine block copolymer, polyethylene glycol-poly (L-aspartic acid derivative) block copolymer, polyethylene glycol-poly (lactic acid)-polyethylene glycol block copolymer, poly (lactic acid)-polyethylene glycol-poly (lactic acid) block copolymer, polyethylene glycol-poly (DL-lactic acid) block copolymer, poly (glycolide-co-lactide)-polyethylene glycol-poly (glycolide-co-lactide) block copolymer, polyethylene glycol-poly (glycolide-co-lactide)-polyethylene glycol block copolymer, polycaprolactone-polyethylene glycol-polycaprolactone block copolymer, polyethylene oxide, polydimethylsiloxane, methacrylate, polyacrylic acid, poly (N-isopropyl acrylamide), 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.

[0012] Preferably, the injectable gel matrix comprises one or more of carbomer, polycarbophil, gelatin, pectin, carrageenan, gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulose polysaccharide, ethylene polymer, acrylic resin, polyvinyl alcohol and polycarboxyvinyl.

[0013] Preferably, the ion-responsive gel matrix comprises one or more of sodium alginate, potassium alginate, calcium alginate, lithium alginate, oxidized sodium alginate, oxidized potassium alginate, oxidized calcium alginate, oxidized lithium alginate, calcium carbonate, calcium oxide, calcium phosphate, calcium heparin, calcium acetate, calcium gluconate, calcium lactate, calcium citrate, and calcium chloride.

[0014] Preferably, the mass ratio of the polymer material and the ultrasound phase transition imaging agent is 1: (1-10).

[0015] Preferably, the polymer material comprises one or more of polylactic acid-glycolic acid copolymer, polyethylene glycol monomethyl ether-polylactic acid-glycolic acid copolymer, polyethylene glycol monomethyl ether-polylactic acid-glycolic acid-polylysine block copolymer, polyethylene glycol-poly-L-aspartic acid derivative block copolymer, polyethylene glycol-poly-lactic acid-polyethylene glycol block copolymer, poly-lactic acid-polyethylene glycol-poly-lactic acid block copolymer, polyethylene glycol-poly-DL-lactic acid block copolymer, polyglycolide-lactide-polyethylene glycol-polyglycolide block copolymer, polyethylene glycol-polyglycolide-lactide-polyethylene glycol block copolymer, polycaprolactone-polyethylene glycol-polycaprolactone block copolymer, lecithin, soybean phospholipid, hydrogenated soybean lecithin, dipalmitoyl phosphatidylcholine, distearoyl phosphatidylcholine, dimyristoyl phosphatidylcholine, dioleoyl lecithin, dipalmitoyl phosphatidyl ethanolamine, distearoyl phosphatidyl ethanolamine, distearoyl phosphatidyl ethanolamine-polyethylene glycol, dipalmitoyl phosphatidyl glycerol, and distearoyl phosphatidyl glycerol.

[0016] Preferably, the ultrasound phase transition imaging agent comprises one or more of perfluoromethane, perfluoroethane, perfluoropropane, perfluorobutane, perfluoropentane, perfluorocyclobutane, perfluorohexane, perfluorooctyl bromide, perfluorocrown ether, sulfur hexafluoride, sodium bicarbonate, ammonium bicarbonate, nitric oxide, and hydrogen.

[0017] Preferably, the pharmaceutically acceptable solvent comprises embryo culture solution.

[0018] The present application also provides a preparation method of the ultrasound imaging composite gel as described above, comprising the following steps:

[0019] The polymer material, the ultrasound phase transition imaging agent, and the pharmaceutically acceptable solvent are first mixed to obtain an ultrasound imaging microbubble suspension;

[0020] The ultrasound imaging microbubble suspension and the gel matrix are second mixed to obtain the ultrasound imaging composite gel.

[0021] Preferably, the first mixing comprises mixing the polymer material and the ultrasound phase transition imaging agent, then sequentially performing emulsification, removing the organic phase, and drying, and then mixing with the pharmaceutically acceptable solvent.

[0022] The application further provides application of the ultrasonic imaging composite gel in the technical solution or the ultrasonic imaging composite gel prepared by the preparation method in the technical solution in preparation of an auxiliary embryo transplantation and implantation preparation.

[0023] The application provides an ultrasonic imaging composite gel, which comprises a gel matrix, an ultrasonic imaging material and a pharmaceutically acceptable solvent according to mass fractions; 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 comprises one or more of a temperature-sensitive gel matrix, an injectable gel matrix and an ion-responsive gel matrix; the ultrasonic imaging material comprises an ultrasonic phase change imaging agent and a high polymer material wrapping the ultrasonic phase change imaging agent, and the ultrasonic imaging material is an ultrasonic imaging microbubble structure. The imaging microbubble in the ultrasonic imaging composite gel has an enhanced ultrasonic imaging function under ultrasonic triggering, which facilitates accurate observation of the embryo implantation process. In addition, compared with the existing embryo culture solution which is a flowing liquid and cannot stably attach the embryo to a specific endometrial site, the embryo migration may have a high risk of ectopic pregnancy. The ultrasonic imaging composite gel is in a semi-solid gel state in the uterine cavity and adheres to the endometrium, which limits the movement of the embryo in the uterine cavity to a certain extent, increases the contact opportunity of the embryo with the endometrium and is thus beneficial to embryo implantation. Therefore, the imaging microbubble in the ultrasonic imaging composite gel can facilitate accurate observation of the embryo implantation process and effectively adhere the embryo to a specific endometrial site. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 An ultrasonic imaging diagram of the ultrasonic imaging microbubble suspension solution described in Embodiment 1 and Embodiments 25-27;

[0025] Figure 2 A gelation condition of the ultrasonic imaging composite gel solution described in Embodiments 1-3 in a thermostat at 37 DEG C;

[0026] Figure 3 An injectable condition of the ultrasonic imaging composite gel described in Embodiments 4-6;

[0027] Figure 4 An imaging effect of local tissues after intramuscular injection of the ultrasonic imaging composite gel described in Embodiment 2, Embodiment 5, Embodiment 7, Embodiment 11, Embodiment 15, Embodiment 16, Embodiment 20 and Embodiment 24;

[0028] Figure 5 A farrowing graph of a control group, Embodiment 2, Embodiment 5, Embodiment 7, Embodiment 11, Embodiment 15, Embodiment 16, Embodiment 20 and Embodiment 24;

[0029] Figure 6 Figure 1 is a graph showing the number of offspring for the control group, Example 2, Example 5, Example 7, Example 11, Example 15, Example 16, Example 20, and Example 24. DETAILED DESCRIPTION

[0030] The present application provides an ultrasonic imaging composite gel, which comprises a gel matrix, an ultrasonic imaging material, and a pharmaceutically acceptable solvent, according to mass fraction.

[0031] The concentration of the gel matrix in the ultrasonic imaging composite gel is 1-500 mg / mL.

[0032] The concentration of the ultrasonic imaging material in the ultrasonic imaging composite gel is 1-500 mg / mL.

[0033] The gel matrix comprises one or more of a temperature-sensitive gel matrix, an injectable gel matrix, and an ion-responsive gel matrix.

[0034] The ultrasonic imaging material comprises an ultrasonic phase transition imaging agent and a polymer material encapsulating the ultrasonic phase transition imaging agent, and the ultrasonic imaging material is an ultrasonic imaging microbubble structure.

[0035] In the present application, all the raw materials for preparation are commercially available products well known to those skilled in the art, unless otherwise specified.

[0036] In the present application, the ultrasonic imaging composite gel preferably exists in the form of a solution or a semi-solid gel.

[0037] In the present 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.

[0038] In the present application, the gel matrix includes one or several of a temperature-sensitive gel matrix, an injectable gel matrix, and an ion-responsive gel matrix; the temperature-sensitive gel matrix includes one or several of poloxamer 407 (F127), poloxamer 188 (F68), polyethylene glycol monomethyl ether-poly(lactic-co-glycolic acid) (mPEG-PLGA), polyethylene glycol monomethyl ether-poly(lactic-co-glycolic acid)-polylysine block copolymer (mPEG-PLGA-PLL), polyethylene glycol-poly-L-aspartic acid derivative block copolymer (PEG-Pasp), polyethylene glycol-poly(lactic acid)-polyethylene glycol block copolymer (PEG-PLA-PEG), poly(lactic acid)-polyethylene glycol-poly(lactic acid) block copolymer (PLA-PEG-PLA), polyethylene glycol-poly(DL-lactic acid) block copolymer (mPEG-PDLLA), polyglycolide-lactide-polyglycolide block copolymer (PLGA-PEG-PLGA), polyethylene glycol-polyglycolide-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(methacrylic acid-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, and more preferably includes poloxamer 407 (F127), poloxamer 188 (F68), carboxymethyl chitosan, or carboxymethyl cellulose. The injectable gel matrix preferably includes one or several of carbomer, polycarbophil, gelatin, pectin, carrageenan, gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulose polysaccharide, ethylene polymer, acrylic resin, polyvinyl alcohol, and polyvinyl acetate, and more preferably includes carbomer; the ion-responsive gel matrix preferably includes one or several of sodium alginate, potassium alginate, calcium alginate, lithium alginate, oxidized sodium alginate, oxidized potassium alginate, oxidized calcium alginate, oxidized lithium alginate, calcium carbonate, calcium oxide, calcium phosphate, heparin calcium, calcium acetate, calcium gluconate, calcium lactate, calcium citrate, and calcium chloride, and more preferably includes sodium alginate, calcium alginate, oxidized calcium alginate, or heparin calcium. In the present application, when the gel matrix is two or more of the above-mentioned specific choices, the present application does not have any special limitation on the ratio of the above-mentioned specific substances, and mixing in any ratio is possible.

[0039] In the present application, the gel matrix can be gelled in the uterus, and the gelling promotes the adhesion of the uterine cavity after the gel is formed.

[0040] In the present 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.

[0041] In the present application, the ultrasonic imaging material includes an ultrasonic phase transition imaging agent and a high molecular material encapsulating the ultrasonic phase transition imaging agent, and the ultrasonic imaging material is an ultrasonic imaging microbubble structure. In the present application, the mass ratio of the high molecular material to the ultrasonic phase transition imaging agent is preferably 1:(1-10), more preferably 1:(5-10), and most preferably 1:10.

[0042] In the present application, the high molecular material preferably includes one or more of polylactic acid-glycolic acid copolymer (PLGA), 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-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-poly-DL-lactic acid block copolymer (mPEG-PDLLA), polyglycolide-lactide-polyethylene glycol-lactide block copolymer (PLGA-PEG-PLGA), polyethylene glycol-polyglycolide-lactide-polyethylene glycol block copolymer (PEG-PLGA-PEG), polycaprolactone-polyethylene glycol-polycaprolactone block copolymer (PCL-PEG-PCL), lecithin, soybean phospholipid, hydrogenated soybean lecithin, dipalmitoyl phosphatidylcholine, distearoyl phosphatidylcholine, dimyristoyl phosphatidylcholine, dioleoyl lecithin, dipalmitoyl phosphatidyl ethanolamine, distearoyl phosphatidyl ethanolamine, distearoyl phosphatidyl ethanolamine-polyethylene glycol, dipalmitoyl phosphatidyl glycerol, and distearoyl phosphatidyl glycerol, and more preferably includes polyethylene glycol monomethyl ether-polylactic acid-glycolic acid-polylysine block copolymer (mPEG-PLGA-PLL) or polylactic acid-glycolic acid copolymer (PLGA). When the high molecular material is two or more of the above specific choices, the present application does not have any special limitation on the ratio of the above specific substances, and any ratio can be mixed.

[0043] In the present application, the ultrasound phase transition imaging agent preferably comprises 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 comprises perfluorohexane or sodium bicarbonate; when the ultrasound phase transition imaging agent is two or more of the above specific choices, the present application does not have any special limitation on the ratio of the above specific substances, and mixing in any ratio can be used.

[0044] In the present application, the ultrasound imaging material is used for embryo transfer ultrasound imaging.

[0045] In the present application, the pharmaceutically acceptable solvent comprises an embryo culture solution. In the present application, the pharmaceutically acceptable solvent also preferably comprises one or more of water for injection, physiological saline, phosphate buffered saline, glucose solution and enzyme-free water, and more preferably water for injection or physiological saline; when the pharmaceutically acceptable solvent is two or more of the above specific choices, the present application does not have any special limitation on the ratio of the above specific substances, and mixing in any ratio can be used. In the present application, the pharmaceutically acceptable solvent serves to disperse the above gel matrix.

[0046] In the present application, the existing flowable embryo culture solution cannot form an in-situ nutrient reservoir, and the loss of nutrients is not conducive to the pre-implantation nutrition supply of embryos. The present application uses an embryo culture solution as a gel solvent, and forms a gel in the uterine cavity to continuously provide early nutrients for embryos, thereby promoting embryo implantation, so that the nutrients (embryo culture solution) in the gel can provide an effective nutrient reservoir for early embryo implantation.

[0047] The present application also provides a preparation method of the ultrasound imaging composite gel according to the above technical solution, comprising the following steps:

[0048] The high molecular material, the ultrasound phase transition imaging agent and the pharmaceutically acceptable solvent are first mixed to obtain an ultrasound imaging microbubble suspension;

[0049] The ultrasound imaging microbubble suspension and the gel matrix are secondly mixed to obtain the ultrasound imaging composite gel.

[0050] The present application first mixes a high molecular material, an ultrasound phase transition imaging agent and a pharmaceutically acceptable solvent to obtain an ultrasound imaging microbubble suspension.

[0051] In the present application, the first mixing preferably comprises mixing the high molecular material and the ultrasound phase transition imaging agent, then sequentially performing emulsification, removing the organic phase and drying, and then mixing with the pharmaceutically acceptable solvent.

[0052] In the present application, the emulsification preferably comprises mixing the mixture of the high molecular material and the ultrasonic phase transition imaging agent with dichloromethane, and then adding an emulsifier and performing ultrasonic treatment. The present application does not have any special limitation on the amount of dichloromethane, the type of emulsifier, the amount of emulsifier, and the conditions of ultrasonic treatment, and the skilled in the art can perform the same. In the embodiments of the present application, the emulsification process can be as follows: 10 mg of high molecular material and 20 mg of ultrasonic phase transition imaging agent are mixed with 500 μL of dichloromethane, and 1.5 mL of 1% polyvinyl alcohol aqueous solution is added for ultrasonic emulsification.

[0053] In the present application, the process of removing the organic phase is preferably vacuum removal, and the temperature of the vacuum removal is preferably room temperature. The present application does not have any special limitation on the specific process of vacuum removal, and the skilled in the art can perform the same.

[0054] In the present application, the drying method is preferably freeze-drying, and the present application does not have any special limitation on the process of freeze-drying, and the skilled in the art can perform the same.

[0055] After obtaining the ultrasonic imaging microbubble suspension, the present application secondarily mixes the ultrasonic imaging microbubble suspension and the gel matrix to obtain the ultrasonic imaging composite gel.

[0056] In the present application, the second mixing is preferably performed under stirring, and the present application does not have any special limitation on the process of stirring, and the skilled in the art can perform the same.

[0057] After the second mixing is completed, when the pH of the obtained mixture does not meet the neutral or reaches the requirement similar to the pH value (7.35-7.45) of the human body, the pH is preferably adjusted by a pH adjusting agent, and the present application does not have any special limitation on the process of adjustment, and the skilled in the art can perform the same.

[0058] The present application also provides the use of the ultrasonic imaging composite gel in the above technical solution or the ultrasonic imaging composite gel prepared by the preparation method in the above technical solution in the preparation of an auxiliary embryo transfer and implantation preparation.

[0059] In the present application, the use preferably comprises adding a perfect embryo in vitro into the ultrasonic imaging composite gel, and implanting the prepared ultrasonic imaging composite gel containing the embryo into the uterus under the guidance of abdominal ultrasound.

[0060] The ultrasonic imaging microbubbles in the ultrasonic imaging composite gel facilitate timing and positioning of an embryo transfer operation under the guidance of an abdominal ultrasound, and the semi-solid adhesion of the ultrasonic imaging composite gel can effectively adhere the embryo to a specific endometrial site, reduce ectopic pregnancy, and promote embryo implantation.

[0061] The technical solutions in the present application will be described clearly and completely below in combination with the embodiments in the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by a person of ordinary skill in the art without creative work fall within the protection scope of the present application.

[0062] Embodiment 1

[0063] 10 mg of polyethylene glycol monomethyl ether-poly-lactic acid-glycolic acid-polylysine block copolymer (mPEG-PLGA-PLL) and 200 μL of perfluorohexane were dissolved in 500 μL of dichloromethane, 1.5 mL of a polyvinyl alcohol aqueous solution with a mass concentration of 1% was added for ultrasonic emulsification. Then, the organic phase was removed under vacuum at room temperature, and freeze-drying was performed, the obtained ultrasonic imaging microbubbles were mixed with 1 mL of water for injection for dissolution, and an ultrasonic imaging microbubble suspension was obtained.

[0064] 150 mg of poloxamer 407 was added to the ultrasonic imaging microbubble suspension, and the mixture was fully stirred and swelled in an ice water bath until completely dissolved, to obtain an ultrasonic imaging composite gel solution.

[0065] Embodiment 2

[0066] With reference to Embodiment 1, the difference is that the amount of poloxamer 407 is replaced by 170 mg.

[0067] Embodiment 3

[0068] With reference to Embodiment 1, the difference is that the amount of poloxamer 407 is replaced by 200 mg.

[0069] The ultrasonic imaging composite gel solutions prepared in Embodiments 1-3 were placed in a thermostat at 37°C, and the gelation of the solutions was observed by the small bottle inversion method, wherein Figure 2 The gelation of the ultrasonic imaging composite gel solutions in Embodiments 1-3 in the thermostat at 37°C was observed by the small bottle inversion method, wherein Figure 2 It can be seen that the ultrasonic imaging composite gel solutions in Embodiments 1-3 can all gelate.

[0070] Embodiment 4

[0071] An ultrasonic imaging microbubble suspension was prepared according to Embodiment 1;

[0072] 5mg of Carbopol was added into the ultrasound imaging microbubble suspension, and after being fully stirred and swelled, the pH value was adjusted to neutral with 1 mol / L sodium hydroxide aqueous solution to obtain the ultrasound imaging composite gel.

[0073] Example 5

[0074] With reference to Example 4, the difference is that the amount of Carbopol is adjusted to 10mg.

[0075] Example 6

[0076] With reference to Example 4, the difference is that the amount of Carbopol is adjusted to 20mg.

[0077] The ultrasound imaging composite gels described in Examples 4-6 were placed in a 20 gauge syringe, and their injectability was observed, wherein Figure 3 The injectability of the ultrasound imaging composite gels described in Examples 4-6 was known from Figure 3 It can be seen that the ultrasound imaging composite gels described in Examples 4-6 are all injectable.

[0078] Example 7

[0079] An ultrasound imaging microbubble suspension was prepared according to Example 1;

[0080] 5mg of sodium alginate was added into the ultrasound imaging microbubble suspension, and after being fully stirred and swelled, 5mg of calcium alginate was added and stirred uniformly to obtain the ultrasound imaging composite gel.

[0081] Example 8

[0082] With reference to Example 7, the difference is that the amount of calcium alginate is replaced by 10mg.

[0083] Example 9

[0084] With reference to Example 7, the difference is that the amount of calcium alginate is replaced by 15mg.

[0085] Example 10

[0086] With reference to Example 7, the difference is that the amount of sodium alginate is replaced by 10mg.

[0087] Example 11

[0088] With reference to Example 7, the difference is that the amount of sodium alginate is replaced by 10mg, and the amount of calcium alginate is replaced by 10mg.

[0089] Example 12

[0090] With reference to Example 7, the difference is that the amount of sodium alginate is replaced by 10mg, and the amount of calcium alginate is replaced by 15mg.

[0091] Example 13

[0092] Reference to Example 7, except that the amount of sodium alginate is replaced by 15 mg.

[0093] Example 14

[0094] Reference to Example 7, except that the amount of sodium alginate is replaced by 15 mg, and the amount of calcium alginate is replaced by 10 mg.

[0095] Example 15

[0096] Reference to Example 7, except that the amount of sodium alginate is replaced by 15 mg, and the amount of calcium alginate is replaced by 15 mg.

[0097] The ultrasonic imaging composite gel of Examples 7-15 is observed by the small bottle inversion method to observe the gelation. At room temperature, the gelation is observed every certain time interval. If there is no flowing gel liquid after inversion, the time is the gelation time. The test results are shown in Table 1:

[0098] Table 1 Gelation performance and gelation time of the ultrasonic imaging composite gel of Examples 7-15

[0099] Example 7 8 9 10 11 12 13 14 15 Gelation Gelation Gelation Gelation Gelation Gelation Gelation Gelation Gelation Gelation Gelation time (min) 28 15 7 17 8 5 9 3 1

[0100] As shown in Table 1, the ultrasonic imaging composite gel of Examples 7-15 can all gelate. Among them, the gelation time of Example 15 is the shortest. Therefore, with the increase of the concentration of sodium alginate and calcium alginate, the gelation time of the ultrasonic imaging composite gel is shortened.

[0101] Example 16

[0102] The ultrasonic imaging microbubble suspension is prepared according to Example 1;

[0103] 10 mg of oxidized sodium alginate is added to the ultrasonic imaging microbubble suspension, and after being fully stirred and swelled, 500 AXaIU of heparin injection is added and stirred uniformly to obtain an ultrasonic imaging composite gel.

[0104] Example 17

[0105] Reference to Example 16, except that the amount of heparin injection is adjusted to 750 AXaIU.

[0106] Example 18

[0107] Reference to Example 16, except that the amount of heparin injection is adjusted to 1000 AXaIU.

[0108] Example 19

[0109] Reference Example 16, except that the amount of oxidized sodium alginate was adjusted to 20 mg.

[0110] Example 20

[0111] Reference Example 16, except that the amount of oxidized sodium alginate was adjusted to 20 mg, and the amount of heparin injection was adjusted to 750 AXaIU.

[0112] Example 21

[0113] Reference Example 16, except that the amount of oxidized sodium alginate was adjusted to 20 mg, and the amount of heparin injection was adjusted to 1000 AXaIU.

[0114] Example 22

[0115] Reference Example 16, except that the amount of oxidized sodium alginate was adjusted to 30 mg.

[0116] Example 23

[0117] Reference Example 16, except that the amount of oxidized sodium alginate was adjusted to 30 mg, and the amount of heparin injection was adjusted to 750 AXaIU.

[0118] Example 24

[0119] Reference Example 16, except that the amount of oxidized sodium alginate was adjusted to 30 mg, and the amount of heparin injection was adjusted to 1000 AXaIU.

[0120] The ultrasonic imaging composite gels described in Examples 16 to 24 were observed for gelation by the vial inversion method. The gels were observed for gelation at room temperature at intervals of a certain time, and the time at which no flowable gel liquid was observed after inversion was taken as the gelation time. The test results are shown in Table 2.

[0121] Table 2 Gelation properties and gelation times of the ultrasonic imaging composite gels described in Examples 16 to 24

[0122] Example 16 17 18 19 20 21 22 23 24 Gelation Gelation Gelation Gelation Gelation Gelation Gelation Gelation Gelation Gelation Gelation time (min) 15 8 5 5 <1 <1 2 <1 <1

[0123] As can be seen from Table 2, the ultrasonic imaging composite gels described in Examples 16 to 23 all gelled, and the gelation time of Example 28 was the shortest. Thus, it can be seen that as the concentrations of oxidized sodium alginate and heparin calcium increase, the gelation time of the ultrasonic imaging composite gel decreases.

[0124] Example 25

[0125] Reference Example 1, except that the polyethylene glycol monomethyl ether-poly(lactic-co-glycolic acid)-polylysine block copolymer was replaced by poly(lactic-co-glycolic acid) copolymer.

[0126] Example 26

[0127] Reference Example 1, except that 200 μL of perfluorohexane was replaced with 20 mg of ammonium bicarbonate.

[0128] Example 27

[0129] Reference Example 25, except that 200 μL of perfluorohexane was replaced with 20 mg of ammonium bicarbonate.

[0130] The ultrasound imaging microbubble suspensions described in Examples 1, 25-27 were tested for in vitro ultrasound imaging enhancement performance: the ultrasound imaging microbubble suspensions described in Examples 1, 25-27 were filled into latex glove finger ends, ultrasound coupling agent was applied to the surface of the latex gloves, and in vitro ultrasound imaging was observed on an ultrasound imaging instrument, wherein Figure 1 The ultrasound imaging images of the ultrasound imaging microbubble suspensions described in Examples 1, 25-27 are shown in Figure 1 It can be seen that the ultrasound imaging microbubble suspensions described in Examples 1, 25-27 can all enhance ultrasound imaging in vitro, and the ultrasound imaging performance of Example 1 is more excellent.

[0131] Test Example

[0132] The ultrasound imaging composite gels described in Examples 2, 5, 7, 11, 15, 16, 20, and 24 were respectively observed for local tissue imaging effects after intramuscular injection, wherein Figure 4 The local tissue imaging effects of the ultrasound imaging composite gels described in Examples 2, 5, 7, 11, 15, 16, 20, and 24 after intramuscular injection are shown in Figure 4 It can be seen that the ultrasound imaging composite gels described in the present application can enhance ultrasound imaging characteristics in vivo;

[0133] 6-8 week old female pseudopregnant fourth day mice were randomly divided into 9 groups, one group of 3, the back was opened, the oviduct was taken out, the oviduct opening part from the ovary was sent into the ampulla part with the oviduct opening part, the negative control group was injected with 10 mouse two-cell embryos containing culture solution, the experimental group was injected with 10 mouse two-cell embryos treated with the ultrasound imaging composite gels described in Examples 2, 5, 7, 11, 15, 16, 20, and 24, respectively, and the number of offspring was observed, wherein Figure 5 The number of offspring of the control group, Examples 2, 5, 7, 11, 15, 16, 20, and 24 is shown in Figure 6A quantitative statistics chart of the number of offspring of the control group, Example 2, Example 5, Example 7, Example 11, Example 15, Example 16, Example 20 and Example 24 is shown in the following figure, wherein Figures 5-6 It can be seen that the ultrasonic imaging composite gel has the effect of promoting assisted embryo implantation, and the offspring rate of Example 20 is the highest.

[0134] The above only describes the preferred embodiments of the present application, and does not limit the present application in any form. It should be noted that for ordinary skilled persons in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should be considered as the protection scope of the present application.

Claims

1. A uterine-gelatinizable ultrasound-detectable composite gel, characterized in that, By weight, it includes the gel matrix, ultrasonic imaging material, 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 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. The gel matrix includes poloxamer 407 or carbomer, or the gel matrix is ​​sodium oxidized alginate and heparin injection, or the gel matrix is ​​sodium alginate and calcium alginate. The ultrasonic phase change imaging agent is perfluorohexane; The polymer material is a polyethylene glycol monomethyl ether-polylactic acid glycol glycolic acid-polylysine block copolymer.

2. 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).

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

4. The method for preparing the ultrasonic imaging composite gel according to any one of claims 1 to 3, 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.

5. The use of the ultrasound imaging composite gel according to any one of claims 1 to 3 or the ultrasound imaging composite gel prepared by the preparation method according to claim 4 in the preparation of assisted embryo transfer and implantation preparations.

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