Polymer, functional preparation, application of functional preparation, preparation method of polymer and preparation method of functional preparation

Abstract

The invention discloses a polymer, a functional preparation, application of the functional preparation, a preparation method of the polymer and a preparation method of the functional preparation. Wherein a polymer single chain in the polymer comprises a hydrophilic chain segment and a hydrophobic chain segment, and a hydrophilic chain link in the hydrophilic chain segment is conjugated with polyethylene glycol; part of polymer single chains are embedded with functional chain links, and at least part of the functional chain links are conjugated with a fluorescent agent; in the polymer, the average number of repetitive units of polyethylene glycol is 4-8, the average molecular weight of the hydrophobic end of the polymer single chain is 1800-7800, and the sum of the average molecular weights of the hydrophobic end and the hydrophilic end of the polymer single chain is 16800-27100. The polymer is prepared through active free radical reaction. The functional preparation is prepared by dispersing the polymer in water and is used for fluorescence imaging of the biliary tract. Compared with the prior art, on the premise that the biliary tract is noninvasive, the phenomenon that the biliary tract cannot be subjected to clear fluorescence imaging can be reduced, or a material basis or a preparation method is provided for achieving the effect.

Description

Technical Field

[0001] This application relates to the field of nanobiomaterials, specifically to a polymer, a functional formulation, the use of the functional formulation, a method for preparing the polymer, and a method for preparing the functional formulation. Background Technology

[0002] With the improvement of living standards, the incidence of biliary tract diseases is increasing year by year. For example, the incidence of gallstones has exceeded 10%. Among gallstone patients, more than 10% require surgical treatment. Because bile ducts in the biliary tract are prone to variation (the variation rate of bile ducts exceeds 50%), bile duct damage can easily occur during surgery due to misjudgment of the shape and direction of the bile ducts. Since some minor bile duct injuries are difficult to detect on laparoscopy, and the bile duct wall is difficult to heal after the wound closes, iatrogenic bile duct injury is common. Patients may need secondary or even multiple surgeries, causing great suffering and even loss of working ability.

[0003] To address this issue, intraoperative navigation is required for biliary tract surgeries. Intraoperative navigation techniques primarily include intraperitoneal ultrasound imaging, endoscopic retrograde cholangiopancreatography (ERCP), and percutaneous transhepatic cholangiopancreatography (PTC).

[0004] In particular, when using abdominal ultrasound imaging, intestinal gas and omentum tissue can significantly interfere with the imaging results, requiring a high level of skill and expertise from the operating physician.

[0005] When ERCP is used, a guidewire is inserted into the common bile duct under X-ray guidance through the opening of the duodenal papilla and iodine contrast agent is injected. ERCP requires trauma to the opening of the duodenal papilla, which carries the risk of intestinal perforation and intraoperative infection. At the same time, ERCP is not suitable for conditions such as acute cholecystitis.

[0006] When using PTC (percutaneous transluminal contrast treatment), a needle with a plastic sheath or a Chiba fine needle is used. The contrast agent is injected into the common bile duct through the right mid-axillary line or anterior approach, under X-ray or ultrasound guidance. PTC carries a risk of failure and inevitably causes trauma to the bile duct, potentially leading to bile leakage. Furthermore, PTC is not suitable for conditions such as purulent biliary obstruction. All of the above techniques for delivering contrast agents into the bile duct cause trauma to the bile duct.

[0007] Furthermore, fluorescence-guided laparoscopic surgery has been widely used in liver surgeries such as hepatectomy. Currently, the fluorescent agent used in fluorescence laparoscopy is indocyanine green. Due to the liver's metabolic properties, indocyanine green is rarely used in biliary tract surgeries.

[0008] Chinese invention patent application CN115887702A (202211473802.5) discloses a nano-fluorescent probe for delivery to the gallbladder, its preparation method, and its application. This nano-fluorescent probe first forms an amphiphilic polymer by polymerizing hydrophilic segments of a brush-like polyethylene glycol chain and hydrophobic segments intercalated with functional segments containing a fluorescent agent. This amphiphilic polymer contains a metal chelate for extending the emission wavelength of the fluorescent agent. The amphiphilic polymer is then dispersed in an isotonic solution to form the nano-fluorescent probe. According to this patent document, the aforementioned nano-fluorescent probe enters the bloodstream via intravenous injection and is rapidly metabolized by the liver into the bile duct, thereby achieving the goal of non-invasively delivering a fluorescent agent into the bile duct and enabling clear fluorescence imaging of the bile duct.

[0009] However, when the applicant studied amphiphilic polymers that do not contain metal chelates according to the guidelines given in the patent document, he found that the nanofluorescent probes that do not contain metal chelates sometimes have very good effects, i.e., clear fluorescence imaging of the bile duct, while at other times there is a phenomenon of liver fluorescence signal halo leading to unclear bile duct imaging, and sometimes they cannot even effectively enable bile duct fluorescence imaging. Summary of the Invention

[0010] The purpose of this application is to overcome the aforementioned defects or problems in the prior art and to provide a polymer, a functional formulation, the use of the functional formulation, a method for preparing the polymer, and a method for preparing the functional formulation. Compared with the prior art, this method, without invasiveness to the bile duct, can reduce the phenomenon of unclear fluorescence imaging of the bile duct, or provide a material basis or preparation method for achieving the above-mentioned effects.

[0011] To achieve the above objectives, the following technical solution is adopted:

[0012] The first technical solution relates to a polymer, which is a collection of polymer single chains; the polymer single chain includes at least one hydrophilic segment and at least one hydrophobic segment of block copolymerization; the hydrophilic segment has a brush-like structure and includes multiple hydrophilic segments, the hydrophilic segments being conjugated with polyethylene glycol; the hydrophobic segment includes multiple hydrophobic segments, the hydrophobic segments being conjugated with hydrophobic groups; at least a portion of the polymer single chain also includes functional segments embedded in the hydrophilic segment and / or the hydrophobic segment; the functional segment is defined as a segment conjugated with a fluorescent agent or a substance formed after the fluorescent agent deactivates, at least a portion of the functional segments being conjugated with a fluorescent agent; in the polymer, polyethylene glycol... The average number of repeating units is between 4 and 8; the first molecular weight of the polymer is between 1800 and 7800; wherein, the first molecular weight of the polymer is defined as the average molecular weight of the hydrophobic ends of the polymer single chains in the polymer, and the hydrophobic ends of the polymer single chains are defined as all hydrophobic segments including the same polymer single chain; the second molecular weight of the polymer is between 16800 and 27100; wherein, the second molecular weight of the polymer is defined as the sum of the average molecular weight of the hydrophobic ends of the polymer single chains in the polymer and the average molecular weight of the hydrophilic ends of the polymer single chains, and the hydrophilic ends of the polymer single chains are defined as all hydrophilic segments including the same polymer single chain.

[0013] The second technical solution is based on the first technical solution, wherein the average number of repeating units of polyethylene glycol in the polymer is between 5 and 8.

[0014] The third technical solution is based on the first technical solution, wherein the molar ratio of the fluorescent agent to the polymer single chain is preferably between 1:20 and 1:80.

[0015] The fourth technical solution is based on the first technical solution, wherein the polymeric groups used to form polymer single chains are groups suitable for undergoing living radical polymerization reactions.

[0016] The fifth technical solution is based on the fourth technical solution, wherein the polymeric group is an acrylate group and / or a methacrylate group.

[0017] The sixth technical solution is based on the first technical solution, wherein the hydrophobic group is any one or more of methyl, ethyl, butyl, hexyl, cyclohexyl, dodecyl and octadecyl.

[0018] The seventh technical solution is based on the sixth technical solution, wherein the hydrophobic group is preferably any one or more of butyl, hexyl, cyclohexyl, and dodecyl.

[0019] The eighth technical solution is based on the first technical solution, wherein the fluorescent agent is any one or more of indocyanine green or its derivatives, boron fluoride pyrrole or its derivatives, IR-783 or its derivatives, IR-820 or its derivatives, FD-1080 or its derivatives, IR-1060 or its derivatives, Cy7 or its derivatives, and Cy5.5 or its derivatives.

[0020] The ninth technical solution is based on the eighth technical solution, wherein the fluorescent agent is preferably any one or more of IR-783 or its derivatives, IR-820 or its derivatives, and indocyanine green or its derivatives.

[0021] The tenth technical solution is based on the first technical solution, wherein the topological structure of the polymer single chain is star-shaped, each arm of the star has both hydrophobic and hydrophilic segments, and on the same arm, the hydrophilic segments are further away from the center of the star than the hydrophobic segments.

[0022] The eleventh technical solution is based on the first technical solution, wherein the topological structure of the polymer single chain is linear.

[0023] The twelfth technical solution is based on the eleventh technical solution, wherein the polymer single chain includes a hydrophilic segment and a hydrophobic segment.

[0024] The thirteenth technical solution is based on the eleventh technical solution, wherein the polymer single chain sequentially includes a first hydrophilic segment, a hydrophobic segment, and a second hydrophilic segment, and both the first hydrophilic segment and the second hydrophilic segment are hydrophilic segments.

[0025] The fourteenth technical solution is based on the eleventh technical solution, wherein the polymer single chain sequentially includes a first hydrophobic segment, a hydrophilic segment, and a second hydrophobic segment, and both the first hydrophobic segment and the second hydrophobic segment are hydrophobic segments.

[0026] The fifteenth technical solution is based on any one of the first to fourteenth technical solutions, wherein at least a portion of the polymer single chain further includes a segment to be modified embedded in the hydrophilic segment and / or the hydrophobic segment, the segment to be modified being conjugated with a first reactive group, the first reactive group being used to conjugate the fluorescent agent through a substitution reaction or a coupling reaction.

[0027] The sixteenth technical solution is based on the fifteenth technical solution, wherein the first reactive group is any one or more of succinimide ester, maleimide ester, azide group, alkynyl group, amino group, mercapto group, hydroxyl group and aldehyde group.

[0028] The seventeenth technical solution is based on the sixteenth technical solution, wherein the first reactive group is preferably a succinimide ester.

[0029] The eighteenth technical solution is based on the fifteenth technical solution, wherein the molar ratio of the first reactive group to the polymer single chain is preferably between 2:1 and 1:3.

[0030] The nineteenth technical solution relates to a functional formulation comprising polymer hydrates dispersed in water or an aqueous solution, wherein at least a portion of the polymer hydrates comprises a polymer as described in any one of the first to eighteenth technical solutions.

[0031] The twentieth technical solution is based on the nineteenth technical solution, wherein the polymer hydrate groups are dispersed in an isotonic solution.

[0032] The twenty-first technical solution is based on the twentieth technical solution, wherein the isotonic solution is physiological saline, glucose injection, Ringer's solution, balanced salt solution or balanced buffer solution.

[0033] The twenty-second technical solution is based on the twentieth technical solution, wherein the average particle size of the polymer hydrate groups in the functional formulation is between 30 and 400 nanometers.

[0034] The twenty-third technical solution relates to the use of a functional preparation, wherein the functional preparation described in any one of the twenty to twenty-two technical solutions is used to deliver a fluorescent agent into the bile duct via blood to perform fluorescence imaging of the bile duct or to provide navigation for fluorescence laparoscopic bile duct surgery by means of fluorescence imaging of the bile duct.

[0035] The twenty-fourth technical solution is based on the twenty-third technical solution, wherein the functional preparation enters the bloodstream via intravenous injection or intravenous infusion; at least a portion of the fluorescent agent reaches the bile duct within 45 minutes after entering the vein.

[0036] The 25th technical solution is based on the 23rd technical solution, wherein the fluorescent agent has a residence time in the bile duct of greater than or equal to 3 hours.

[0037] The twenty-sixth technical solution is based on the twenty-third technical solution, wherein the functional preparation enables clear fluorescence imaging of the bile duct for a time greater than or equal to 2 hours, and the clear fluorescence imaging of the bile duct refers to the fluorescence signal intensity of the bile duct and / or gallbladder being significantly greater than the fluorescence signal intensity of the liver.

[0038] The twenty-seventh technical solution relates to the use of a functional preparation, wherein the functional preparation described in any one of the twenty to twenty-two technical solutions is used to deliver a fluorescent agent to the digestive tract downstream of the bile duct via blood to perform fluorescence imaging of the digestive tract downstream of the bile duct, the digestive tract downstream of the bile duct including the duodenum, small intestine and colon.

[0039] The twenty-eighth technical solution relates to a method for preparing a polymer, which is used to prepare the polymer as described in any one of the first to fourteenth technical solutions, wherein the preparation method includes: the polymer being polymerized by a living free radical polymerization reaction; the hydrophilic segment being polymerized by a hydrophilic monomer, wherein the hydrophilic monomer forms the hydrophilic link in the hydrophilic segment; the hydrophobic segment being polymerized by a hydrophobic monomer, wherein the hydrophobic monomer forms the hydrophobic link in the hydrophobic segment; and a functional monomer conjugated with a fluorescent agent being inserted into the corresponding segment during the formation of at least one segment, wherein the functional monomer forms the functional link after being inserted.

[0040] The twenty-ninth technical solution is based on the twenty-eighth technical solution, wherein the functional monomer is prepared by a substitution reaction or a coupling reaction of a first unit and a second unit, the first unit includes a polymeric group and a first reactive group connected to each other, and the second unit includes a fluorescent agent and a second reactive group suitable for coupling reaction or substitution reaction with the first reactive group connected to each other.

[0041] The thirtieth technical solution relates to a method for preparing a polymer, which is used to prepare a polymer as described in any one of the fifteenth to eighteenth technical solutions. The preparation method includes: the polymer being polymerized by a living free radical polymerization reaction; the hydrophilic segment being polymerized by a hydrophilic monomer, wherein the hydrophilic monomer forms the hydrophilic chain segment in the hydrophilic segment; the hydrophobic segment being polymerized by a hydrophobic monomer, wherein the hydrophobic monomer forms the hydrophobic chain segment in the hydrophobic segment; a monomer to be modified, conjugated with a first reactive group, being inserted into a corresponding segment during the formation of at least one segment; the monomer to be modified forming a pre-modified chain segment after insertion; and a fluorescent agent being allowed to probabilistically modify the pre-modified chain segment to form a functional chain segment under reaction conditions suitable for substitution or coupling reactions, wherein the unmodified fluorescent agent pre-modified chain segment forms the modified chain segment.

[0042] The thirty-first technical solution is based on the thirty-first technical solution, wherein the fluorescent agent and the second reactive group are connected to form a second unit, and the second unit and the prepared chain segment to be modified are formed into the functional chain segment through a probabilistic substitution reaction or coupling reaction between the second reactive group and the first reactive group.

[0043] The thirty-second technical solution is based on any one of the twenty-eighth to thirty-first technical solutions, wherein the average number of repeating units of polyethylene glycol in the polymer is controlled by controlling the average number of repeating units of polyethylene glycol in all hydrophilic monomers; the first molecular weight of the polymer is controlled by controlling the degree of polymerization of the hydrophobic monomers; and the second molecular weight of the polymer is controlled by controlling the degree of polymerization of the hydrophobic monomers and the degree of polymerization of the hydrophilic monomers, respectively.

[0044] The thirty-third technical solution is based on the thirty-second technical solution, wherein the degree of polymerization of the hydrophobic monomer is controlled by controlling the molar ratio of the hydrophobic monomer to the chain transfer agent or initiator and the conversion rate of the living free radical polymerization reaction used to polymerize the hydrophobic segment; the degree of polymerization of the hydrophilic monomer is controlled by controlling the molar ratio of the hydrophilic monomer to the chain transfer agent or initiator and the conversion rate of the living free radical polymerization reaction used to polymerize the hydrophilic segment.

[0045] The thirty-fourth technical solution relates to a method for preparing a functional preparation, which is used to prepare a functional preparation as described in the nineteenth technical solution, wherein the functional preparation is prepared by dispersing a polymer as described in any one of the first to eighteenth technical solutions in water or an aqueous solution.

[0046] Compared with existing technologies, the above solution has the following beneficial effects:

[0047] As described in the background section of this application, the applicant, based on the parameter range indicated in the published patent application CN115887702A, has indeed verified that, in some cases, the nanofluorescent probes can be rapidly metabolized by the liver and enter the bile duct, enabling clear fluorescence imaging of the bile duct. The applicant believes that the most significant technical contribution of the technical solution described in this patent document is the discovery that nanofluorescent probes prepared from the relevant amphiphilic polymers can indeed be rapidly metabolized by the liver and enter the bile duct, enabling clear fluorescence imaging of the bile duct.

[0048] However, the applicant also discovered that when using the parameter range indicated in the aforementioned patent documents to study amphiphilic polymers and nano-fluorescent probes without metal chelates, there were sometimes instances of liver fluorescence signal haloing leading to unclear fluorescence imaging of the bile duct, and sometimes there were instances where bile duct fluorescence imaging could not be effectively achieved. Since the aforementioned patent documents did not explain the mechanism by which the defined nano-fluorescent probes can rapidly enter the bile duct via liver metabolism and achieve clear fluorescence imaging of the bile duct, nor did they indicate which parameters are related to the realization or non-realization of this mechanism, the applicant, through continuous experimentation, analysis, and research, intends to reveal this mechanism, determine which parameters are related to the realization of this mechanism, and determine the parameter range that enables the realization of this mechanism.

[0049] This application is based on research into this mechanism and related parameters. Compared to the aforementioned patent literature, the applicant's research reveals more clearly the mechanism by which the nanofluorescent probes prepared from the relevant amphiphilic polymers can rapidly enter the bile duct via hepatic metabolism and achieve clear fluorescence imaging of the bile duct. Furthermore, compared to existing technologies, the applicant's research clarifies the parameter range that enables this mechanism. While the research into the relevant mechanism and the determined parameter range were obtained during research on amphiphilic polymers and nanofluorescent probes without metal chelates, these mechanisms and parameter ranges are also of substantial significance for amphiphilic polymers and nanofluorescent probes containing metal chelates. Therefore, this application does not exclude amphiphilic polymers and nanofluorescent probes containing metal chelates from the scope of protection of this application. The parameter range defined in this application, compared to the parameter range given in the aforementioned patent literature, better ensures that amphiphilic polymers containing metal chelates can achieve clear fluorescence imaging of the bile duct without invasiveness.

[0050] The applicant's research revealed that the reason why the aforementioned fluorescent nanoprobes cause liver fluorescence signal haloing, resulting in unclear fluorescence imaging of the bile duct, is that in some cases, a significant portion of the fluorescent nanoprobes are phagocytosed by hepatocytes and enter the bile duct via exocytosis. Furthermore, if the fluorescent nanoprobes are phagocytosed by hepatocytes and enter the bile duct via exocytosis, it not only causes liver fluorescence signal haloing, resulting in unclear fluorescence imaging of the bile duct, but also leads to significant individual differences in the time it takes for the fluorescent nanoprobes to reach the bile duct, with differences reaching the order of several hours. This variability will cause difficulties in practical application, making it impossible for surgeons performing fluorescence laparoscopic surgery to schedule patients' surgical times.

[0051] The applicant's research revealed that the reason why the aforementioned nano-fluorescent probes cannot effectively perform biliary fluorescence imaging is that, in some cases, a significant portion of the nano-fluorescent probes are metabolized by the kidneys, resulting in the observation of strong fluorescence signals in the bladder, while insufficient nano-fluorescent probes are metabolized by the liver and enter the bile duct. The applicant also observed that, in some cases, the aforementioned nano-fluorescent probes cannot disperse in water or aqueous solutions, which also prevents them from effectively performing biliary fluorescence imaging.

[0052] In summary, the applicant's research found that the mechanism by which the above-mentioned nanofluorescent probes can be rapidly metabolized by the liver and enter the bile duct to enable clear fluorescence imaging of the bile duct is that, provided they can be dispersed in water or aqueous solution, only a small or very small amount is metabolized by the kidneys and only a small or very small amount is phagocytosed by hepatocytes, thereby enabling them to rapidly reach the bile duct via liver metabolism and enable clear fluorescence imaging of the bile duct.

[0053] The applicant's main difficulty during the research process lay in determining which parameters affected whether the fluorescent nanoprobes were metabolized by the kidneys and which parameters affected whether they were phagocytosed by hepatocytes. Of course, the applicant also had to identify which parameters affected the dispersion of the fluorescent nanoprobes in water or aqueous solutions. Since the main technical contribution of this patent application lies in discovering the parameters that influence the realization of the mechanism and in ensuring the realization of the relevant mechanism more effectively than existing technologies through parameter selection, the applicant will describe the arduous process of discovering each of these parameters.

[0054] First, discovering the parameters that affect whether the nanofluorescent probes are phagocytosed by hepatocytes is the most difficult part.

[0055] Existing literature records that amphiphilic polymers, when introduced into water or aqueous solutions, self-assemble to form polymer hydrates. These polymer hydrates contain a large number of polymer single chains. Based on the teachings of the prior art, polymer hydrates cannot be dissociated by simple liquid pressure. Existing technical literature also records that in blood circulation, polymer hydrates may dissociate due to shear forces between them and the blood vessel walls, proteins in the blood, or blood cells. This dissociation property should not and cannot occur extensively in the polymer hydrates formed by the amphiphilic polymers involved in this application; otherwise, the polymer hydrates, after dissociation in the blood vessels, would be easily metabolized by the kidneys and would not be able to enter the bile ducts in large quantities via liver metabolism.

[0056] According to literature, the normal pathway for the fluorescent probe to enter the bile duct via liver metabolism includes hepatocytes and the intercellular spaces. If it is not phagocytosed by hepatocytes, it can only pass through the intercellular spaces, which are described in the literature as being approximately 3 nanometers in size. However, the amphiphilic polymers in the aforementioned patent literature have hydrated groups with particle sizes ranging from tens to hundreds of nanometers. Therefore, those skilled in the art, including the applicant, cannot correlate the particle size of the polymer hydrated groups with whether the fluorescent probe rapidly enters the bile duct via liver metabolism. They can only consider that other factors may cause the aforementioned fluorescent probe to rapidly enter the bile duct via liver metabolism.

[0057] After numerous experiments, the applicant found no other factors related to whether the aforementioned fluorescent probe was rapidly metabolized by the liver and entered the bile duct. Therefore, they turned their attention to the correlation between the particle size of the amphiphilic polymer hydrate clusters and whether it was rapidly metabolized by the liver and entered the bile duct. However, experiments confirmed that the particle size of the amphiphilic polymer hydrate clusters was not closely related to whether it was rapidly metabolized by the liver and entered the bile duct. Some experiments showed that even with larger polymer hydrate cluster particle sizes, rapid metabolization by the liver and entry into the bile duct were still possible; other experiments showed that even with smaller polymer hydrate cluster particle sizes, they could still be phagocytosed by hepatocytes and enter the bile duct via exocytosis.

[0058] After exploring all possibilities, the applicant creatively hypothesized that the aforementioned polymer hydrates might not dissociate in blood vessels, but only in the hepatic sinusoids of the liver. To verify this hypothesis, the applicant innovatively used molecular dynamics simulation software to calculate the size of polymer single-chain hydrates formed by the self-assembly of polymer single chains in water under various parameters. This calculation was time-consuming, labor-intensive, and unprecedented. Combining the calculation results with a large amount of relevant experimental data, it was finally discovered that the size of the polymer single chains in water is strongly correlated with whether the nano-fluorescent probes are rapidly metabolized by the liver and enter the bile duct. That is, if the calculated particle size of the polymer single chains in water is less than 3 nanometers in the intercellular space of hepatocytes, there is a high probability that they can be rapidly metabolized by the liver and enter the bile duct; if the calculated particle size of the polymer single chains in water is greater than or equal to 3 nanometers in the intercellular space of hepatocytes, they are easily phagocytosed by hepatocytes and enter the bile duct via exocytosis. Thus, the applicant discovered a mechanism of action never before documented in existing literature: the aforementioned polymer hydrates that cannot be dissociated in blood vessels can be at least partially dissociated into polymer single-chain hydrates under the special structure of the hepatic sinusoids, thereby enabling them to rapidly pass through the gaps between hepatocytes to reach the bile duct.

[0059] However, the size of the polymer single chain in water is only a result of software calculations. These calculations require a large number of parameters. Existing literature does not explicitly teach which parameters of this type of amphiphilic polymer single chain significantly affect its size in water. Current technology only roughly indicates a positive correlation between the size of the polymer single chain in water and the molecular weight of the polymer single chain within the polymer. Therefore, the applicant conducted numerous experiments and found that the molecular weight of the polymer single chain shows a correlation with its size in water, but this cannot be considered the sole characteristic. Software calculations and experimental results indicate that even some polymer single chains with relatively large molecular weights may still have small sizes in water, allowing for rapid liver metabolism and entry into the bile duct. Other experiments show that even polymer single chains with relatively small molecular weights may still have large sizes in water, preventing rapid liver metabolism and entry into the bile duct.

[0060] Based on the aforementioned software calculations and experimental results, the applicant creatively proposes that in this type of polymer structure, the molecular weight of the hydrophobic end of the polymer single chain and the sum of the molecular weights of the hydrophobic and hydrophilic ends of the polymer single chain jointly determine the size of the hydrate of the amphiphilic polymer single chain, thereby affecting whether the relevant fluorescent agent can be rapidly metabolized by the liver and enter the bile duct. Here, the hydrophobic end refers to all hydrophobic segments of the same polymer single chain, and the hydrophilic end refers to all hydrophilic segments of the same polymer single chain.

[0061] However, further experiments by the applicant revealed that even if the average molecular weight of the hydrophobic ends of the polymer single chains in the polymer (hereinafter referred to as "first molecular weight") and the sum of the average molecular weight of the hydrophobic ends of the polymer single chains in the polymer and the average molecular weight of the hydrophilic ends of the polymer single chains in the polymer (hereinafter referred to as "second molecular weight") are appropriate, there are still other factors that affect whether the nanofluorescent probe is phagocytosed by hepatocytes.

[0062] The applicant continuously searched and designed experiments among various parameters, and finally creatively discovered that the number of repeating units of polyethylene glycol in the hydrophilic chain also affects whether the fluorescent nanoprobe is phagocytosed by hepatocytes. When the average number of repeating units of polyethylene glycol in the polymer is greater than 8 or less than 4, even if the first and second molecular weights are appropriate, the fluorescent nanoprobe will still be phagocytosed by hepatocytes. No related experimental phenomena have been reported in the literature.

[0063] Thus, the key parameters affecting whether the fluorescent nanoprobe is phagocytosed by hepatocytes and enters the bile duct via exocytosis have been revealed. The applicant has determined the parameter range in which the fluorescent nanoprobe is not easily phagocytosed by hepatocytes in large quantities. Specifically, when the average number of repeating units of polyethylene glycol in the polymer is between 4 and 8, and the first molecular weight of the polymer is less than or equal to 7800 and the second molecular weight of the polymer is less than or equal to 27100, the fluorescent nanoprobe is not easily phagocytosed by hepatocytes in large quantities.

[0064] Secondly, discovering the parameters affecting whether nanofluorescent probes are metabolized by the kidneys is also very difficult. Existing literature records that substances metabolized by the kidneys are generally smaller than 8 to 13 nanometers, while the particle size of polymer hydrates is between tens and hundreds of nanometers, much larger than substances typically metabolized by the kidneys. The kidneys also lack special structures like hepatic sinusoids that allow polymer hydrates to dissociate into hydrated polymer single chains. As mentioned above, existing technical literature describes that in blood circulation, polymer hydrates may dissociate due to shear forces generated with blood vessel walls, proteins in the blood, or blood cells. This dissociation is related to the strength of the binding forces between hydrophobic segments in the amphiphilic polymer. Based on the above literature, the applicant designed an experiment to verify whether the first molecular weight of the polymer and other parameters related to the hydrophobic ends of the polymer single chains are related to renal metabolism. Analysis of the experimental results revealed that the first molecular weight of the polymer single chain only showed a certain correlation with whether it was metabolized by the kidneys, but there was no strong correlation. Some experiments have shown that when the first molecular weight of the polymer is too small, there is indeed a problem with renal metabolism; other experiments have shown that when the first molecular weight of the polymer is determined, there is sometimes a problem with renal metabolism, and sometimes there is no problem with renal metabolism. Therefore, the applicant believes that there are other parameters that affect whether the nanofluorescent probe is metabolized by the kidneys.

[0065] The applicant, while searching for parameters influencing whether fluorescent nanoprobes are phagocytosed by hepatocytes, discovered a correlation between the polymer's second molecular weight and whether it is phagocytosed. The applicant creatively hypothesized that the polymer's second molecular weight is also related to whether it is metabolized by the kidneys. To this end, the applicant designed and verified experiments, finding that the polymer's first and second molecular weights are indeed key parameters in determining whether fluorescent nanoprobes are metabolized by the kidneys.

[0066] Thus, the key parameters affecting whether the fluorescent nanoprobe is metabolized by the kidneys have been revealed. The applicant has determined the parameter range within which the fluorescent nanoprobe is not easily metabolized by the kidneys in large quantities; that is, when the first molecular weight is greater than or equal to 1800 and the second molecular weight is greater than or equal to 16800, the fluorescent nanoprobe is not easily metabolized by the kidneys in large quantities.

[0067] Finally, the applicant hypothesized that the key parameter affecting the dispersibility of the fluorescent nanoprobes lies in the average number of repeating units of polyethylene glycol (PEG) in the polymer. Based on this hypothesis, the applicant designed an experiment, and the experimental results verified the hypothesis: the key parameter affecting the water solubility of the fluorescent nanoprobes is indeed the average number of repeating units of PEG in the polymer. When the average number of repeating units of PEG is 4, special methods are required to disperse the polymer in water or aqueous solutions; when the average number of repeating units of PEG is greater than 3 but less than 4, the dispersibility of the fluorescent nanoprobes is poor, and some fluorescent nanoprobes cannot be dispersed; when the average number of repeating units of PEG is less than or equal to 3, at least the vast majority of the fluorescent nanoprobes cannot be dispersed in water or aqueous solutions.

[0068] Thus, the key parameters that can reduce the phenomenon of unclear fluorescence imaging of the bile duct have been revealed. This result is described in the first technical solution, namely, when the average number of repeating units of polyethylene glycol in the polymer is between 4 and 8, the first molecular weight of the polymer is between 1800 and 7800, and the second molecular weight of the polymer is between 16800 and 27100, the nanofluorescent probe is more likely to achieve the goal of rapid liver metabolism to reach the bile duct and enable clear fluorescence imaging of the bile duct compared with the prior art.

[0069] In the first technical solution, the applicant, through experiments, expanded the topological structure of the polymer single chain. Firstly, when the polymer single chain is linear, the number of hydrophilic and hydrophobic segments, as well as the order of block copolymerization, are not important. Whether it's one hydrophilic segment copolymerized with one hydrophobic segment, two hydrophilic segments copolymerized on either side of a hydrophobic segment, or two hydrophobic segments copolymerized on either side of a hydrophilic segment, experiments have shown that the same technical effect can be achieved. The key lies in the average number of repeating units of polyethylene glycol in the polymer, the polymer's first molecular weight, and the polymer's second molecular weight. Secondly, the applicant, through experiments, discovered that even if the polymer single chain exhibits a star-shaped or other topological structure, as long as the hydrophilic segments can be located on the outer side and the hydrophobic segments can be curled up inside the hydrophilic segments after the polymer is immersed in water or an aqueous solution, various topological structures are permissible.

[0070] In the first technical solution, the applicant has verified through experiments that the functional link is a feasible technical solution whether it is embedded in a hydrophilic link segment or a hydrophobic link segment.

[0071] In the first technical solution, "block copolymerization" implies random copolymerization excluding hydrophilic and hydrophobic linkages. Since block copolymerization, under current technical conditions, can only be achieved through living radical polymerization and anionic polymerization, both of which are polymerization reactions with highly controllable molecular weight distribution, "block copolymerization" implicitly implies that the molecular weight of the hydrophobic end and the total molecular weight of the hydrophobic and hydrophilic ends have distribution characteristics comparable to those of living radical polymerization and anionic polymerization. In fact, the applicant also prepared the relevant polymers through living radical polymerization in its experiments, and the experimental results were based on the premise that the molecular weight of the hydrophobic end and the total molecular weight of the hydrophobic and hydrophilic ends have distribution characteristics comparable to those of living radical polymerization and anionic polymerization. The applicant did not use anionic polymerization in its experiments only because its preparation cost is very high and the reaction conditions are very harsh. Those skilled in the art will know that relevant polymers with the same degree of polymerization and molecular weight distribution can also be prepared through anionic polymerization. Of course, the relevant hydrophobic groups, primary reactive groups, etc., need to be selected to meet the requirements of anionic polymerization reactions.

[0072] In the first technical solution, the polyethylene glycol on the hydrophilic segments has the function of resisting protein adsorption. However, the applicant found that only when the average number of repeating units of polyethylene glycol is between 4 and 8 can it avoid being massively phagocytosed by hepatocytes. Of course, the non-toxicity of polyethylene glycol to mammals has been confirmed. At the same time, the structural characteristics of polyethylene glycol itself enable the hydrophilic segments to exhibit a brush-like structure.

[0073] In the first technical solution, the fact that the functional chain segment can be conjugated with the substance formed after the fluorescent agent fails means that when a functional monomer conjugated with a fluorescent agent participates in the polymerization reaction, some of the fluorescent agent fails due to the destruction of the conjugated chain caused by the high temperature of the polymerization reaction.

[0074] In both the first and second technical solutions, "intercalation" includes both block copolymerization and random copolymerization. However, since functional segments and segments to be modified are scarce in polymer single chains, random copolymerization is generally sufficient, resulting in higher preparation efficiency.

[0075] In the second technical solution, an average number of repeating units of polyethylene glycol between 5 and 8 is a more preferred technical solution. The polymer has better dispersibility in water or aqueous solution, is less likely to be phagocytosed by liver cells, and is less likely to be metabolized by the kidneys.

[0076] The preferred molar ratio of fluorescent agent to polymer single chain is 1:20 to 1:80, which can effectively avoid the quenching effect when there is too much fluorescent agent.

[0077] In the fourth technical solution, "polymeric groups used to form polymer single chains" refers to the polymeric groups used to form the main chain of the polymer single chain. These polymeric groups are suitable for undergoing living radical polymerization. Essentially, they control the molecular weight and distribution of the hydrophobic ends of the polymer single chain, as well as the total molecular weight and distribution of the hydrophobic and hydrophilic ends, through living radical reactions. Experiments have shown that living radical reactions can completely achieve the required distribution. Furthermore, the cost of living radical polymerization is significantly lower than other alternative anionic polymerization reactions.

[0078] The fifth technical solution is the preferred embodiment of the polymeric group verified by the applicant through experiments. In the polymer, acrylate groups and methacrylate groups can be used alone or in combination.

[0079] The sixth technical solution is a preferred embodiment of the hydrophobic group, as verified by the applicant through experiments and well-known to those skilled in the art. It should be noted that the hydrophobic group here refers not only to a group possessing hydrophobic properties, but also to a hydrophobic group capable of twisting or turning in three-dimensional space, so that different polymer single chains interweave, entangle, or interpenetrate each other during self-assembly in water or aqueous solutions, giving the polymer hydrates a certain binding force. This hydrophobic group can be linear or ring-shaped. Linear hydrophobic groups can interweave and entangle with each other. Ring-shaped hydrophobic groups can interpenetrate with each other.

[0080] In the seventh technical solution, methyl and ethyl groups are preferably not used as hydrophobic groups because the applicant found during the experiment that while methyl and ethyl groups have a possibility of success as hydrophobic groups under the condition that other parameters are the same, they also have a possibility of failure. Analysis revealed that this is mainly because when methyl and ethyl groups are used as hydrophobic groups, the hydrophobic chain segments are relatively short, and the interlacing and entanglement bonding forces are low, resulting in a certain degree of dispersion in the bonding force of the polymer hydrate groups.

[0081] The eighth and ninth technical solutions involve the selection of fluorescent agents, wherein the fluorescent agents are preferably one or more of IR-783 or its derivatives, IR-820 or its derivatives, and indocyanine green or its derivatives, which are easier to modify through substitution or coupling reactions, resulting in higher reaction efficiency and lower cost.

[0082] In the tenth technical solution, the polymer single chain has a star-shaped topological structure, which is one of the optional topological structures. On the same arm, the hydrophilic segment is further away from the center of the star than the hydrophobic segment, so as to better utilize the amphiphilic properties of the polymer. This makes the polyethylene glycol on the outside less susceptible to phagocytosis by hepatocytes.

[0083] In technical solutions eleven through fourteen, the linear topology of the polymer single chain is the preferred topology. Using a single chain results in the lowest preparation cost. The technical solution employing one hydrophilic segment and one hydrophobic segment not only has low cost but also higher preparation efficiency.

[0084] In the fifteenth technical solution, the segment to be modified is used to probabilistically modify it with a fluorescent agent, thereby converting it into a functional segment. As is well known to those skilled in the art, this modification can be a substitution reaction between a first reactive group and a second reactive group, or a coupling reaction between the first reactive group and the second reactive group. Those skilled in the art will understand that whether the segment to be modified is embedded in a hydrophilic segment or a hydrophobic segment, it is a feasible technical solution.

[0085] The first reactive group listed in the sixteenth technical solution is a group well-known to those skilled in the art that is suitable for substitution or coupling reactions. The applicant has excluded the carboxyl group as the first reactive group, mainly because purification is very difficult when the carboxyl group is used as the first reactive group.

[0086] In the seventeenth technical solution, the first reactive group is preferably succinimide ester, which has milder reaction conditions, higher reaction efficiency, and makes it easier for fluorescent agents to be modified on the prepared chain segments.

[0087] In the eighteenth technical solution, the molar ratio of the first reactive group to the polymer is preferably 2:1 to 1:3. If combined with the third technical solution, where the molar ratio of the fluorescent agent to the polymer is preferably 1:20 to 1:80, the number of functional and unmodified segments on the polymer single chain is very small. Therefore, the functional and unmodified segments have little impact on the properties of the polymer and the realization of related mechanisms, and can be ignored. More importantly, the first reactive group of the unmodified segment and the fluorescent agent of the functional segment both possess planar characteristics, making it difficult for them to interweave, entangle, or interpenetrate with hydrophobic segments or other functional or unmodified segments. Therefore, the contribution of the unmodified and functional segments to the properties of the functional formulation is very small and can be ignored.

[0088] The nineteenth to twenty-first technical solutions are functional formulations formed by dispersing the above-mentioned polymers in water or aqueous solutions. The aqueous solution, if isotonic, is suitable for injection into the bloodstream. It is known to those skilled in the art that amphiphilic polymers self-assemble in water to form polymer hydrates. A polymer hydrate contains a polymer, meaning that the polymer hydrate contains a large number of polymer single chains. In water, the hydrophilic ends (mainly hydrophilic segments) of the polymer single chains open outwards, while their hydrophobic ends curl inwards. Even if the functional chain segments are located in the hydrophilic segments, due to the brush-like structure of the hydrophilic segments, the fluorescent agent can still be "hidden" in the outwardly opening "bristles" formed by polyethylene glycol, effectively preventing the fluorescent agent from being phagocytosed by Kupffer cells, hepatic sinusoidal endothelial cells, and hepatocytes.

[0089] In the 20th and 21st technical solutions, the aqueous solution is an isotonic solution, which has good biocompatibility, low toxicity, and is suitable for injection into the blood, and has great potential for clinical application.

[0090] The twenty-second technical solution is the measured average particle size range of polymer hydrates.

[0091] Technical solution 23 illustrates the use of the functional formulations defined in technical solutions 20 and 21 in biliary fluorescence imaging and laparoscopic fluorescence biliary surgical navigation. Compared with existing technologies, it can reduce the phenomenon of unclear fluorescence imaging of the biliary tract without invasiveness to the biliary tract.

[0092] The twenty-fourth technical solution illustrates the time it takes for the fluorescent agent to reach the bile duct. It shows that after the functional preparation is injected into the bloodstream, it can be rapidly metabolized by the liver and enter the bile duct. Patients requiring corresponding surgery do not need to wait for an uncertain time, and no individual variability is observed. Therefore, this is highly beneficial for surgical efficiency and practical clinical application.

[0093] The twenty-fifth technical solution illustrates the residence time of the fluorescent agent in the bile duct. Due to the characteristics of bile duct function, the fluorescent agent can remain in the bile duct for a long time.

[0094] In the twenty-sixth technical solution, the functional agent enables clear fluorescence imaging of the bile duct for a time greater than or equal to 2 hours, which is very beneficial for navigation in fluorescence laparoscopic surgery with a certain degree of complexity.

[0095] In the twenty-seventh technical solution, the functional preparation is metabolized through the bile duct and excreted into the downstream digestive tract, enabling fluorescence imaging of the digestive tract downstream of the bile duct.

[0096] The twenty-eighth technical solution illustrates a method for preparing polymers using functional monomers. The key lies in the ability to effectively control the molecular weight and distribution of the hydrophobic ends of polymer single chains, as well as the total molecular weight and distribution of both hydrophobic and hydrophilic ends, through living radical polymerization. However, when preparing functional units from functional monomers, due to the high temperature of the living radical polymerization reaction, some fluorescent agents may fail due to conjugated chain failure. Therefore, the substances conjugated in the functional units may include substances formed after the fluorescent agent's failure, in addition to the fluorescent agent itself. The method of preparing polymers using free radical polymerization is relatively simple and can achieve large-scale production.

[0097] The twenty-ninth technical solution illustrates a method for preparing a functional monomer, which is known to those skilled in the art.

[0098] The thirtieth technical solution illustrates a method for preparing polymers by modifying a fluorescent agent after forming a polymer from a monomer to be modified. The key lies in the ability to effectively control the molecular weight and distribution of the hydrophobic ends of the polymer single chains, as well as the total molecular weight and distribution of the hydrophobic and hydrophilic ends of the polymer single chains, through active free radical reactions. Specifically, when the conditions of the substitution or coupling reaction differ from the polymerization conditions, modification should be performed after the polymerization reaction in which the desired modified segment is inserted into the corresponding chain segment; when the conditions of the substitution or coupling reaction are the same as or partially the same as the polymerization conditions, modification is permissible during the polymerization reaction. The method of preparing polymers using free radical polymerization is relatively simple and can achieve large-scale production.

[0099] The modification method in the thirty-first technical solution is known to those skilled in the art.

[0100] Technical solutions 32 and 33 illustrate methods for controlling the average number of repeating units in polyethylene glycol, the first molecular weight of the polymer, and the second molecular weight of the polymer.

[0101] The thirty-fourth technical solution illustrates a method for preparing a functional formulation, in which a polymer is dispersed in water or an aqueous solution and self-assembles to form polymer hydrates, thereby forming a functional formulation. Attached Figure Description

[0102] To more clearly illustrate the technical solutions of the embodiments, the accompanying drawings used are briefly described below:

[0103] Figure 1 This is a schematic diagram of the polymer single chain structure in Example 1;

[0104] Figure 2 This is a schematic diagram of the polymer hydration group structure in Example 1;

[0105] Figure 3a The image shows a whole-body fluorescence imaging of the skin in mice after intravenous injection of the functional formulation in Example 1.

[0106] Figure 3b These are fluorescence imaging images of the liver and gallbladder after intravenous injection of the functional formulation in Example 1 into mice.

[0107] Figure 4a The image shows a laparoscopic fluorescence imaging of the liver and gallbladder in healthy rabbits after intravenous injection of indocyanine green.

[0108] Figure 4b The images shown are laparoscopic fluorescence images of the liver and gallbladder after intravenous injection of the functional formulation in Example 1 into healthy rabbits.

[0109] Figure 5a Images showing stained liver tissue sections of healthy rabbits after intravenous injection of the functional formulation from Example 1;

[0110] Figure 5b Image of liver tissue sections stained with indocyanine green after intravenous injection into healthy rabbits;

[0111] Figure 6a The image shows a laparoscopic fluorescence imaging of the liver and gallbladder in rabbits after intravenous injection of indocyanine green to induce liver injury.

[0112] Figure 6b The image shows a laparoscopic fluorescence imaging of the liver and gallbladder after intravenous injection of the functional preparation in Example 1 into a rabbit with liver injury.

[0113] Figure 7a Image of liver tissue sections stained with indocyanine green after intravenous injection into rabbits with liver injury;

[0114] Figure 7b Images of stained liver tissue sections from rabbits with liver injury after intravenous injection of the functional formulation described in Example 1.

[0115] Figure 8a This is a laparoscopic fluorescence imaging image of the liver and gallbladder after intravenous injection of indocyanine green into a rabbit with bile obstruction.

[0116] Figure 8b The image shows a laparoscopic fluorescence imaging of the liver and gallbladder after intravenous injection of the functional preparation in Example 1 into a rabbit with bile duct obstruction.

[0117] Figure 9a Image of liver tissue sections stained with indocyanine green after intravenous injection in rabbits with biliary obstruction;

[0118] Figure 9b Images showing stained liver tissue sections of rabbits with biliary obstruction after intravenous injection of the functional formulation from Example 1;

[0119] Figure 10 The image shows a laparoscopic fluorescence imaging of the liver and gallbladder in healthy rabbits after intravenous infusion of the functional formulation described in Example 1.

[0120] Figure 11a The image shows a fluorescently guided image of the functional formulation in Example 1 during the separation of the Greenson sheath in porcine cholecystectomy.

[0121] Figure 11b The image shows a fluorescently guided image of the functional formulation used in Example 1 during the separation of the gallbladder artery in a pig cholecystectomy.

[0122] Figure 11c The image shows a fluorescent navigation image of the functional formulation used in Example 1 during the ligation of the gallbladder artery in a pig cholecystectomy.

[0123] Figure 11d The image shows a fluorescently guided image of the functional formulation used in Example 1 during the severance of the gallbladder artery in a pig cholecystectomy.

[0124] Figure 11e The image shows a fluorescently guided image of the functional formulation used in Example 1 during the separation of the cystic duct in a porcine cholecystectomy.

[0125] Figure 11f The image shows the fluorescent navigation of the functional preparation in Example 1 during the ligation of the cystic duct in pig cholecystectomy.

[0126] Figure 11g The image shows the fluorescent navigation of the functional formulation in Example 1 during the cystic duct transection stage of porcine cholecystectomy.

[0127] Figure 11h The image shows a fluorescently guided image of the functional formulation used in Example 1 during the separation of the gallbladder bed in a porcine cholecystectomy.

[0128] Figure 11i The image shows the fluorescent navigation of the functional formulation in Example 1 during the cholecystectomy stage in pigs.

[0129] Figure 12a A schematic diagram showing the comparison of cell viability of mouse embryonic fibroblasts NIH3T3 in different concentrations of the functional formulation of Example 1;

[0130] Figure 12b A schematic diagram comparing the cell viability of human umbilical vein epithelial cells (HUVECs) in different concentrations of the functional formulation from Example 1.

[0131] Figure 12c A schematic diagram comparing the cell viability of normal human hepatocytes in different concentrations of the functional formulation of Example 1;

[0132] Figure 13a Stained tissue sections of various organs of live mice 14 days after injection of saline.

[0133] Figure 13b Stained images of tissue sections from various organs of live mice 14 days after injection of the functional formulation of Example 1;

[0134] Figure 14 Images of stained sections of reproductive organ tissue from live mice 30 days after injection of the functional formulation of Example 1;

[0135] Figure 15a This is a transdermal whole-body fluorescence imaging image of the functional formulation in mice after intravenous injection in Example 2;

[0136] Figure 15b These are fluorescence imaging images of the liver and gallbladder after intravenous injection of the functional formulation in Example 2 into mice.

[0137] Figure 16a This is a transdermal whole-body fluorescence imaging image of mice after intravenous injection of the functional formulation in Example 3;

[0138] Figure 16b These are fluorescence imaging images of the liver and gallbladder after intravenous injection of the functional formulation in Example 3 into mice.

[0139] Figure 17a This is a transdermal whole-body fluorescence imaging image of the functional formulation in mice after intravenous injection in Example 4;

[0140] Figure 17b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 4.

[0141] Figure 18a This is a transdermal whole-body fluorescence imaging image of the functional formulation in mice after intravenous injection in Example 5;

[0142] Figure 18b These are fluorescence imaging images of the liver and gallbladder after intravenous injection of the functional formulation in Example 5 into mice.

[0143] Figure 19a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 6 after intravenous injection in mice;

[0144] Figure 19b These are fluorescence imaging images of the liver and gallbladder after intravenous injection of the functional formulation in Example 6 into mice.

[0145] Figure 20aThis is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 7 after intravenous injection in mice;

[0146] Figure 20b These are fluorescence imaging images of the liver and gallbladder after intravenous injection of the functional formulation in Example 7 into mice.

[0147] Figure 21a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 8 after intravenous injection in mice;

[0148] Figure 21b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 8.

[0149] Figure 22a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 9 after intravenous injection in mice;

[0150] Figure 22b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 9.

[0151] Figure 23a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 10 after intravenous injection in mice;

[0152] Figure 23b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 10.

[0153] Figure 24a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 11 after intravenous injection in mice;

[0154] Figure 24b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 11.

[0155] Figure 25a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 12 after intravenous injection in mice;

[0156] Figure 25b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 12.

[0157] Figure 26a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 13 after intravenous injection in mice;

[0158] Figure 26b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation from Example Thirteen.

[0159] Figure 27a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 14 after intravenous injection in mice;

[0160] Figure 27b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 14.

[0161] Figure 28a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 15 after intravenous injection in mice;

[0162] Figure 28b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 15.

[0163] Figure 29a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 16 after intravenous injection in mice;

[0164] Figure 29b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example Sixteen.

[0165] Figure 30a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 17 after intravenous injection in mice;

[0166] Figure 30b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation from Example 17.

[0167] Figure 31a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 18 after intravenous injection in mice;

[0168] Figure 31b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 18.

[0169] Figure 32a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 19 after intravenous injection in mice;

[0170] Figure 32b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation from Example 19.

[0171] Figure 33a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 20 after intravenous injection in mice;

[0172] Figure 33b These are fluorescence imaging images of the liver and gallbladder of mice after intravenous injection of the functional formulation in Example 20.

[0173] Figure 34a This is a transdermal whole-body fluorescence imaging image of the functional formulation in Example 21 after intravenous injection in mice;

[0174] Figure 34bThe images shown are fluorescence images of the liver and gallbladder after intravenous injection of the functional formulation in Example 21 into mice.

[0175] Figure 35 This is an image showing the state of the polymer dispersed in water in Comparative Example 1.

[0176] Figure 36 The image shows the state of the polymer dispersed in water in Comparative Example 2.

[0177] Figure 37a Transdermal whole-body fluorescence imaging images of mice after intravenous injection of the functional formulations in Comparative Example 3;

[0178] Figure 37b The images show fluorescence imaging of the liver and gallbladder after intravenous injection of the functional formulations in Comparative Example 3 into mice.

[0179] Figure 38a Transdermal whole-body fluorescence imaging images of mice after intravenous injection of the functional formulations in Comparative Example 4;

[0180] Figure 38b Fluorescence imaging images of the liver and gallbladder after intravenous injection of the functional formulations in Comparative Example 4 into mice.

[0181] Figure 39a Transdermal whole-body fluorescence imaging of mice in a supine position after intravenous injection of the functional formulations in Comparative Example 5.

[0182] Figure 39b Transdermal whole-body fluorescence imaging of mice in a prone position after intravenous injection of the functional formulations in Comparative Example 5;

[0183] Figure 40 Transdermal whole-body fluorescence imaging images of mice after intravenous injection of the functional formulations in Comparative Example 6;

[0184] Figure 41a Transdermal whole-body fluorescence imaging images of mice after intravenous injection of the functional formulations in Comparative Example 7;

[0185] Figure 41b The images show fluorescence imaging of the liver and gallbladder after intravenous injection of the functional formulations in Comparative Example 7 into mice.

[0186] Figure 42a Transdermal whole-body fluorescence imaging images of mice after intravenous injection of the eight functional formulations in comparison examples;

[0187] Figure 42b Fluorescence imaging images of the liver and gallbladder after intravenous injection of the eight functional preparations in mice, serving as comparative examples.

[0188] Figure 43 Transdermal whole-body fluorescence imaging images of mice after intravenous injection of the functional formulations in Comparative Example 9;

[0189] Figure 44a Transdermal whole-body fluorescence imaging images of mice after intravenous injection of the functional formulations of Comparative Example 10.

[0190] Figure 44b Fluorescence imaging images of the liver and gallbladder after intravenous injection of the functional formulations in Comparative Example 10 into mice.

[0191] Figure 45a Transdermal whole-body fluorescence imaging images of mice after intravenous injection of functional formulations in Comparative Example 11;

[0192] Figure 45b Fluorescence imaging of the liver and gallbladder after intravenous injection of the functional formulation in Comparative Example 11 into mice;

[0193] Figure 46 A schematic diagram of the hepatic sinusoidal structure;

[0194] Figure 47 yes Figure 46 A magnified view of part A;

[0195] Figure 48 This is a molecular dynamics simulation diagram of the polymer single chain in water in Example 1;

[0196] Explanation of key figure labels:

[0197] 1. Hydrophilic segment; 2. Hydrophobic segment; 3. Polyethylene glycol; 4. Hydrophobic group; 5. Fluorescent agent; 6. Gallbladder; 7. Liver; 8. Duodenum; 9. Small intestine; 10. Colon; 11. Cecum; 12. Common bile duct; 13. Normal liver area; 14. Damaged liver area; 15. Cystic duct; 16. Bladder; 17. Kidney; 18. Hepatic artery; 19. Portal vein; 20. Hepatic sinusoids; 21. Central vein; 22. Perisinusoidal space; 23. Hepatic sinusoidal endothelial cells; 24. Capillary bile ducts; 25. Hepatocytes; 26. Intrahepatic bile ducts. Detailed Implementation

[0198] Unless otherwise specified, the terms “first,” “second,” or “third,” etc., in the claims and description are used to distinguish different objects and not to describe a particular order.

[0199] Unless otherwise specified, the terms “comprising,” “having,” and variations thereof in the claims and description shall mean “including but not limited to.”

[0200] In the claims and description, unless otherwise specified, the term "have" means that a technical feature that follows is part of a technical feature that precedes it.

[0201] In the claims and specification, unless otherwise specified, the term "polymer" refers to a collection of polymer single chains. That is, a polymer is completely equivalent to a collection of polymer single chains. The phrase "polymer hydrate contains polymer" means that the polymer hydrate contains multiple polymer single chains. In other words, regardless of the number of polymer single chains, the collection is a polymer. In this application, the method for obtaining the polymer from the functional formulation is preferably to dialyze the functional formulation in deionized water and then completely dry it.

[0202] Unless otherwise specified in the claims and description, the term "block copolymer" refers to the linkage of two or more polymer segments with different properties. Specifically, block copolymerization in this application does not include random copolymerization. Whether a polymer is a block copolymer is determined by: hydrogen nuclear magnetic resonance (HMR) spectroscopy of the polymer. Block copolymerization is indicated when the characteristic peaks of the methyl group in the methacrylate or the methylene group in the acrylate exhibit a bimodal distribution; block copolymerization is not indicated when the characteristic peaks of the methyl group in the methacrylate or the methylene group in the acrylate exhibit a single peak or a broad distribution. Where the obtained sample is a functional formulation, the polymer is obtained by dialyzing in deionized water and then completely drying.

[0203] Unless otherwise specified in the claims and description, the term "hydrophobic group" refers not only to a group with hydrophobic functionality, but also to a hydrophobic group capable of twisting or turning in three-dimensional space so that different polymer single chains interweave, entangle, or interpenetrate each other during self-assembly in water or aqueous solutions, thereby giving the polymer hydrates a certain binding force. Such hydrophobic groups can be linear or cyclic. Linear hydrophobic groups can interweave or entangle with each other. Cyclic hydrophobic groups can interpenetrate with each other.

[0204] Unless otherwise specified in the claims and description, the term "intercalation" means that a functional link or a link to be modified is located on a polymer segment, which may include block copolymers or random copolymers, and may be located at the end or the middle of the segment.

[0205] Unless otherwise specified in the claims and specification, the term "substance formed after fluorescent agent failure" refers to the substance formed when a functional monomer conjugated with a fluorescent agent participates in a polymerization reaction, and some of the fluorescent agent fails due to the destruction of its conjugated chain caused by high temperature.

[0206] In the claims and specification, unless otherwise specified, the term "number of repeating units of polyethylene glycol" refers to the amount of ethylene glycol used to form the polyethylene glycol. The term "average number of repeating units of polyethylene glycol in the polymer" refers to the average number of repeating units of polyethylene glycol on all hydrophilic segments of all polymer single chains in the polymer. In this application, the "average number of repeating units of polyethylene glycol in the polymer" is preferably detected by the following method: First, the polymer is hydrolyzed to obtain a mixture of polyethylene glycol, an alcohol with hydrophobic groups, and the main chain of the polymer single chain with water; second, the mixture is placed in dichloromethane to extract the organic components, resulting in a mixture of polyethylene glycol, an alcohol with hydrophobic groups, and the main chain of the polymer single chain with dichloromethane; third, a portion of the dichloromethane is removed, and the remaining mixture is placed in n-hexane and the main chain of the polymer single chain is removed by sedimentation, with both polyethylene glycol and the alcohol with hydrophobic groups remaining in the supernatant; finally, after the supernatant is completely dried, the obtained substance is subjected to high-performance liquid chromatography to obtain substances and molecules with similar molecular weights. The average molecular weight and mass percentage of substances with similar molecular weights were determined. The chemical structure of each substance in the substance with similar molecular weight and its molar ratio to other substances in the same molecular weight were obtained by proton nuclear magnetic resonance spectroscopy. All substances with chemical structures belonging to polyethylene glycol and the number of repeating units in each substance were identified. Based on the number of repeating units in the polyethylene glycol-containing substances, the molar ratio of this substance to other substances with similar molecular weights, the average molecular weight of the substances with similar molecular weights to which this substance belongs, and the mass ratio of the substances with similar molecular weights to other substances with similar molecular weights, the average number of repeating units of polyethylene glycol in the polymer was determined.

[0207] In the claims and description, unless otherwise specified, the term "between" is defined as including both endpoints, i.e., greater than or equal to the smaller endpoint and less than or equal to the larger endpoint.

[0208] Unless otherwise specified in the claims and description, the term "hydrophobic end of a polymer single chain" is defined as all hydrophobic segments of the same polymer single chain.

[0209] Unless otherwise specified in the claims and description, the term "hydrophilic end of a polymer single chain" is defined as all hydrophilic segments of the same polymer single chain.

[0210] Unless otherwise specified in the claims and description, the term "first molecular weight" is defined as the average molecular weight of the hydrophobic ends of the polymer single chains in the polymer. The first molecular weight is accurate to the hundreds place, and values ​​less than one hundred should be rounded up. For example, a theoretical or actual measured value of 2530 is defined as the first molecular weight of 2600.

[0211] Unless otherwise specified in the claims and description, the term "second molecular weight" is defined as the sum of the average molecular weight of the hydrophobic ends and the average molecular weight of the hydrophilic ends of the polymer single chains in the polymer. The second molecular weight is accurate to the hundreds place, and values ​​less than one hundred should be rounded up. For example, if the theoretical or actual measured value is 18030, its second molecular weight is defined as 18100.

[0212] The first and second molecular weights are preferably determined as follows: First, the average molecular weight of the polymer is obtained by gel permeation chromatography; second, the chemical structures of all hydrophobic groups, polymeric groups, the molar ratios of all hydrophobic groups to all polyethylene glycols, and the molar ratios of all polyethylene glycols to all polymeric groups are obtained by proton nuclear magnetic resonance spectroscopy. Based on the average molecular weight of the polymer, the average number of repeating units in the polyethylene glycol, the chemical structures of the hydrophobic groups and polymeric groups, the molar ratios of all hydrophobic groups to all polyethylene glycols, and the molar ratios of all polyethylene glycols to all polymeric groups, the average molecular weight of the hydrophilic ends of all polymer single chains and the first molecular weight are obtained, and then the second molecular weight is obtained. Other methods may be used for determination if they are more accurate and easier.

[0213] Unless otherwise specified in the claims and description, the term "biliary tract" refers to the gallbladder and extrahepatic bile ducts.

[0214] Unless otherwise specified in the claims and description, the term "clear fluorescence imaging of the bile ducts" means that the fluorescence signal intensity of the gallbladder and / or bile ducts is significantly stronger than that of the liver.

[0215] Unless otherwise specified in the claims and description, the term "molar ratio of hydrophilic or hydrophobic monomer to chain transfer agent or initiator" refers to a chain transfer agent when the active radical reaction is a reversible addition-fragmentation chain transfer polymerization (RAFT) and an initiator when the active radical reaction is an atom transfer radical polymerization (ATRP).

[0216] Unless otherwise specified in the claims and description, the term "probabilistic" in "the second unit and the proposed modified linker form the functional linker through a probabilistic substitution or coupling reaction between the second reactive group and the first reactive group" means that the substitution or coupling reaction between the second reactive group and the first reactive group is probabilistic when the first reactive group is located in the proposed modified linker, and does not mean that the substitution or coupling reaction between the second reactive group and the first reactive group is probabilistic under all conditions.

[0217] Unless otherwise specified in the claims and specification, the term "IR-808" refers to one of the derivatives of IR783, specifically, the product obtained by introducing a carboxyl group into IR783 through a substitution reaction. The molecular structure of IR783 is as follows:

[0218]

[0219] The molecular structure of IR-808 is as follows:

[0220]

[0221] The technical solutions in the embodiments will be clearly and completely described below with reference to the accompanying drawings.

[0222] Example 1

[0223] The polymer in Example 1 is a collection of polymer single chains. Each polymer single chain comprises at least one hydrophilic segment and at least one hydrophobic segment of a block copolymer. See also... Figure 1 , Figure 1 The structure of the polymer single chain in Example 1 is shown. Figure 1 As shown in Example 1, the polymer single chain has a linear topological structure. The polymer single chain includes a hydrophilic segment 1 and a hydrophobic segment 2, both from block copolymers. The hydrophilic segment 1 has a brush-like structure and includes multiple hydrophilic links, each conjugated with polyethylene glycol 3. The hydrophobic segment 2 includes multiple hydrophobic links, each conjugated with a hydrophobic group 4. Here, the hydrophobic group 4 refers to a group with hydrophobic functionality, and also refers to a hydrophobic group that can twist or turn in three-dimensional space so that different polymer single chains interweave, entangle, or interpenetrate each other during self-assembly in water or an aqueous solution, giving the polymer hydrates a certain binding force. This hydrophobic group can be linear or ring-shaped. Linear hydrophobic groups can interweave and entangle with each other. Ring-shaped hydrophobic groups can interpenetrate with each other. Figure 1The polymer single chain shown also includes functional segments embedded in the hydrophobic segment 2, and these functional segments are conjugated with fluorescent agent 5. For the polymer, a functional segment is defined as a segment conjugated with fluorescent agent 5 or with a substance formed after the fluorescent agent has deactivated. The substance formed after the fluorescent agent has deactivated refers to the substance formed when a functional monomer conjugated with fluorescent agent 5 participates in the polymerization reaction, and some of the fluorescent agent 5 deactivates due to the destruction of its conjugated chain at high temperatures. Generally, the molar ratio between the substance formed after the fluorescent agent has deactivated and the fluorescent agent 5 is approximately 4:6. The substance formed after the fluorescent agent has deactivated has the same molecular weight as the fluorescent agent 5, and the fluorescent agent 5 has a conjugated planar structure. For the polymer in Example 1, at least a portion of the polymer single chain includes functional segments embedded in the hydrophobic segment 2, and at least a portion of these functional segments are conjugated with fluorescent agent 5. The molar ratio of the functional segment to the polymer single chain is approximately 1:25, and the molar ratio of the fluorescent agent 5 to the polymer single chain is approximately 1:40. In other embodiments, the functional linker may be embedded in the hydrophobic segment 2, the hydrophilic segment 1, or both the hydrophilic segment 1 and the hydrophobic segment 2. In other embodiments, the molar ratio of the fluorescent agent 5 to the polymer single chain is preferably between 1:20 and 1:80.

[0224] In Example 1, the polymer molecular formula is P-OEGMA 50 -b-(LMA 14-co-(MA-IR-808)). OEGMA represents a hydrophilic monomer with an average repeating unit count of approximately 4.5 (the commercially available oligomeric polyethylene glycol methacrylate OEGMA has a nominal average repeating unit count of 5-6 and an average molecular weight of 300; however, testing revealed an actual average repeating unit count of approximately 4.5). LMA represents a hydrophobic monomer with a dodecyl group. MA-IR-808 represents a functional monomer that is a derivative of IR-783 as a fluorescent agent. The preparation process of MA-IR-808 includes a first step of introducing carboxyl groups into IR-783 via a substitution reaction to prepare IR-808, and a second step of introducing polymeric groups into IR-808 via an amidation reaction to prepare MA-IR-808. IR-808 is a derivative of IR-783. The first step in preparing IR-808 is as follows: IR-783 (180 mg, 0.24 mmol) and 4-mercaptobenzoic acid (74 mg, 0.48 mmol) are dissolved in 5 mL of DMF and stirred at room temperature for 24 h under a nitrogen atmosphere. After the reaction is complete, the solution is precipitated in diethyl ether, washed three times with diethyl ether, and thoroughly dried to obtain 193 mg of IR-808, with a yield of 93%. The thiol group (-SH) on the 4-mercaptobenzoic acid undergoes a substitution reaction with the chlorine atom on IR-783. In the second step of preparing MA-IR-808 in this embodiment, the functional monomer MA-IR-808 is obtained from the first unit and the second unit through a probabilistic substitution reaction. The first unit includes a polymeric group and a first reactive group connected to each other; the polymeric group is a methacrylate group, and the first reactive group is an amino group. The second unit is IR-808, which includes a fluorescent agent connected to each other and a second reactive group suitable for substitution reaction with the first reactive group; the fluorescent agent is IR-783, and the second reactive group is a carboxyl group. The substitution reaction was carried out in DMF under weakly alkaline conditions and at room temperature. The substitution reaction occurred with a probability of over 90% during the preparation of the functional monomer. After the reaction, the functional monomer MA-IR-808 was purified by column chromatography. Specifically, in this embodiment, the second step was as follows: IR-808 (100 mg, 0.12 mmol), 2-aminoethyl methacrylate hydrochloride (40 mg, 0.24 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbonyldimethylamine hydrochloride (EDC, 46 mg, 0.24 mmol) were dissolved in 5 mL of DMF and stirred under a nitrogen atmosphere for 30 min. Then, 1-hydroxybenzotriazole (Hobt, 33 mg, 0.24 mmol) was added. The mixture was stirred at 40 °C for 24 h. After the reaction, the solution was placed in a 1 kDa dialysis bag and deionized for 48 h. After lyophilization, 102 mg of MA-IR-808 was obtained, with a yield of 90%.In IR-808, the carboxyl group undergoes an amidation reaction with the amino group in 2-aminoethyl methacrylate hydrochloride, introducing a methacrylate polymer group.

[0225] In Example 1, the polymer groups used for the hydrophilic monomer, hydrophobic monomer, and functional monomer are all methacrylate groups. In the polymer, the average number of hydrophilic segments in the hydrophilic chain is 50. The average number of repeating polyethylene glycol units in the hydrophilic segments is approximately 4.5. The average number of hydrophobic segments in the hydrophobic chain is 14. The hydrophobic group in the hydrophobic segments is dodecyl. On average, there is one functional segment for every 40 polymer single chains. The fluorescent agent conjugated to the functional segment is IR-808, a derivative of IR-783. The substance formed after the fluorescent agent conjugated to the functional segment deactivates is the substance formed after the deactivation of IR-808, a derivative of IR-783. In Example 1, the average molecular weight (hereinafter referred to as "first molecular weight") of the hydrophobic ends of the polymer single chains is 3600, where the hydrophobic ends of the polymer single chains are defined as including all hydrophobic segments of the same polymer single chain. The sum of the average molecular weight of the hydrophobic end and the average molecular weight of the hydrophilic end (hereinafter referred to as "second molecular weight") of the polymer single chain is 18,600, where the hydrophilic end of the polymer single chain is defined as including all hydrophilic segments of the same polymer single chain.

[0226] The preparation method of the polymer in Example 1 is as follows:

[0227] Step 1: Preparation of hydrophilic segments via living radical polymerization:

[0228] In Example 1, the living radical polymerization reaction used was a reversible addition-fragmentation chain transfer (RAFT) polymerization reaction. The chain transfer agent was 4-cyano-4-(thiobenzoyl)valerate (CTA). The hydrophilic monomer was oligomeric polyethylene glycol methacrylate (OEGMA), more specifically polyethylene glycol methyl ether methacrylate, with an average repeating unit count of approximately 4.5 and a polymeric group of methacrylate. The initiator was azobisisobutyronitrile (AIBN). The solvent for calibration by 1H NMR spectroscopy was anisole.

[0229] First, CTA 56 mg (0.2 mmol), OEGMA 3.42 g (11.4 mmol), and AIBN 3.3 mg (0.02 mmol) were dissolved in 15 mL of 1,4-dioxane, and 3 to 4 drops of anisole were added. The mixture was reacted in a Shrek tube, and the conversion rate was monitored by 1H NMR spectroscopy. The reaction conditions were: after deoxygenation, the temperature was raised to 70 °C. During the reaction, an appropriate amount of the mixture was taken and the conversion rate was monitored by 1H NMR spectroscopy. When the conversion rate reached 87.9%, oxygen was introduced to stop the reaction. After the reaction was completed, the mixture was cooled to room temperature, precipitated in n-hexane, the supernatant was removed, and the product was dried under vacuum at room temperature to obtain 2.93 g of hydrophilic segment. The molecular formula of the hydrophilic segment is P-OEGMA. 50 Hydrophilic monomers form hydrophilic chains within hydrophilic segments.

[0230] Step 2: Preparation of polymers via living radical polymerization

[0231] The chain transfer agent for the living radical polymerization reaction is the hydrophilic segment (P-OEGMA) prepared in step 1. 50 The hydrophobic monomer is lauryl methacrylate (LMA, with a dodecyl hydrophobic group and a methacrylate polymeric group). The functional monomer is MA-IR-808 (the fluorescent agent is IR-808, a derivative of IR-783, with a methacrylate polymeric group). The initiator is AIBN. The calibration solvent for 1H NMR monitoring is anisole.

[0232] First, P-OEGMA 50 1.5 g (0.1 mmol), 458 mg (1.8 mmol) LMA, 10 mg MA-IR-808, and 3.3 mg AIBN were dissolved in 15 mL of N,N-dimethylformamide (DMF), and 3 to 4 drops of anisole were added. The mixture was reacted in a Shrek tube, and the conversion was monitored by 1H NMR spectroscopy. The reaction conditions were: deoxygenation followed by heating to 70 °C. During the reaction, a suitable amount of the mixture was taken and the conversion was monitored by 1H NMR spectroscopy. The reaction was stopped when oxygen was introduced when the conversion reached 78.7%. After the reaction, the mixture was cooled to room temperature, the DMF was evaporated to dryness, and then redissolved in 15 mL of tetrahydrofuran. The supernatant was then removed by precipitation in n-hexane, and the polymer was dried under vacuum at room temperature to obtain 1.74 g of polymer. The molecular formula of the polymer is P-OEGMA. 50 -b-(LMA 14 -co-(MA-IR-808)). In this reaction, hydrophobic monomers form hydrophobic linkages within the hydrophobic segments; functional monomers form functional linkages within the hydrophobic segments. During the reaction, due to the high temperature of the polymerization reaction, some of the fluorescent agent becomes inactive. The molar ratio of the substance formed after the fluorescent agent inactive to the fluorescent agent is approximately 4:6.

[0233] In Example 1, the average number of repeating polyethylene glycol units in the polymer is controlled by controlling the average number of repeating units in all hydrophilic monomers. The first molecular weight of the polymer is controlled by controlling the degree of polymerization of the hydrophobic monomer. The second molecular weight of the polymer is controlled by controlling the degree of polymerization of the hydrophobic monomer and the hydrophilic monomer, respectively. Specifically, the degree of polymerization of the hydrophobic monomer is controlled by controlling the molar ratio of the hydrophobic monomer to the chain transfer agent or initiator and the conversion rate of the living radical polymerization reaction used to polymerize the hydrophobic segments; the degree of polymerization of the hydrophilic monomer is controlled by controlling the molar ratio of the hydrophilic monomer to the chain transfer agent or initiator and the conversion rate of the living radical polymerization reaction used to polymerize the hydrophilic segments.

[0234] In other embodiments, the reaction of modifying the fluorescent agent to the polymeric group can also be a coupling reaction. The first reactive group can also be any one or more of succinimide ester, maleimide ester, azide, alkynyl, amino, mercapto, hydroxyl, and aldehyde. When the first reactive group is succinimide ester and the second reactive group is amine, the modification is achieved through a substitution reaction; when the first reactive group is maleimide ester and the second reactive group is thiol, the modification is achieved through a coupling reaction; when the first reactive group is azide and the second reactive group is alkynyl, the modification is achieved through a coupling reaction; when the first reactive group is alkynyl and the second reactive group is azide, the modification is achieved through a coupling reaction; when the first reactive group is amine and the second reactive group is succinimide ester or halogen, the modification is achieved through a substitution reaction; when the first reactive group is thiol and the second reactive group is maleimide ester or halogen, the modification is achieved through a substitution reaction; when the first reactive group is hydroxyl and the second reactive group is carboxyl, the modification is achieved through a substitution reaction or dihydropyran, the modification is achieved through a coupling reaction; when the first reactive group is aldehyde and the second reactive group is amine, the modification is achieved through a coupling reaction.

[0235] The polymer is dispersed in water or an aqueous solution to prepare the functional formulation. In Example 1, the polymer is dispersed in an isotonic solution to prepare the functional formulation. Of course, the polymer can also be dispersed in water, and then an isotonic solution is prepared. The isotonic solution can be physiological saline, glucose injection, Ringer's solution, balanced salt solution, or balanced buffer solution; in this example, a balanced buffer solution is used.

[0236] See Figure 2 , Figure 2 The structure of polymer hydrates in functional formulations is shown. After the polymer is dispersed in water or an aqueous solution, polymer hydrates form through self-assembly in the water or aqueous solution. Functional formulations contain a large number of polymer hydrates. Each polymer hydrate contains a large number of polymer single chains, ranging in size from hundreds to tens of thousands. At least a portion of the polymer hydrate contains polymer single chains with functional repeating segments. Figure 2As shown, in the polymer hydrate, the hydrophilic polyethylene glycol 3 extends outward, while the hydrophobic groups 4 of the hydrophobic groups and the fluorescent agent 5 and the substance formed after the fluorescent agent deactivates curl inward. Because the hydrophobic groups 4 of each polymer single chain have the characteristic of being able to twist or bend in three-dimensional space, when the polymer single chains curl inward, they intertwine, entangle, and interpenetrate with each other in three-dimensional space, giving the polymer hydrate a certain structural strength. This results in a certain degree of stability in water, aqueous solutions, or blood, making it difficult to dissociate. In this embodiment, the average particle size of the polymer hydrate in the functional formulation was measured to be 45.8 nanometers using dynamic light scattering.

[0237] The functional formulation in Example 1 can be used for biliary fluorescence imaging, or for biliary fluorescence imaging to guide laparoscopic surgery of the biliary tract. The functional formulation in Example 1 can also be used for fluorescence imaging of the digestive tract downstream of the bile duct.

[0238] Specifically, the functional formulation in Example 1 was administered via tail vein injection into the bloodstream of mice at a dose of 2 mg of the effective fluorescent agent per kg body weight. See also Figure 3a and Figure 3b , Figure 3a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 1 after intravenous injection in mice; Figure 3b The image shown is a fluorescence imaging image of the liver and gallbladder anatomy in mice after intravenous injection of the functional formulation of Example 1. Figure 3a As shown, in this embodiment, at least a portion of the fluorescent agent exhibits a fluorescent signal in the gallbladder 6 5 minutes after intravenous injection. Figure 3b As shown, in this embodiment, the fluorescent agent remains in the gallbladder 6 for more than 5 hours. Even 5 hours after injection, the fluorescent agent still enables clear fluorescence imaging of the gallbladder 6, meaning the fluorescence intensity of the gallbladder 6 is greater than that of the liver 7. Because the bile ducts of mice are too small, they cannot be clearly distinguished from the gallbladder 6. Fifteen minutes after injection, fluorescent signals appear in the digestive tract downstream of the bile duct. Specifically, the digestive tract downstream of the bile duct mainly includes the duodenum 8, small intestine 9, colon 10, and cecum 11.

[0239] The functional formulation in Example 1 was injected into the bloodstream via the marginal ear vein of healthy rabbits at a dose of 0.5 mg of effective fluorescent agent per kg body weight. See also... Figure 4a and Figure 4b . Figure 4a The image shows a laparoscopic fluorescence imaging of the liver and gallbladder after intravenous injection of commercially available indocyanine green into healthy rabbits. Figure 4b The image shown is a laparoscopic fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 1 into healthy rabbits. Figure 4aAs shown, commercially available indocyanine green produced a fluorescent signal in the bile duct 10 minutes after injection, but fluorescence imaging could not clearly distinguish the liver and bile duct until 90 minutes after injection. Figure 4b As shown, in Example 1, the functional formulation produced a fluorescent signal in the bile duct 3 minutes after injection; from 10 minutes after injection, the fluorescent signals of the common bile duct 12 and gallbladder 6 were stronger than those of the liver 7, indicating that the bile duct was clearly imaged with fluorescence; 20 minutes after injection, the fluorescent signal of the liver 7 had greatly decreased, while the fluorescent signals of the common bile duct 12 and gallbladder 6 were stronger. The fluorescent signals of the common bile duct 12 and gallbladder 6 remained strong until 80 minutes after injection. See also... Figure 5a and Figure 5b . Figure 5a Images of liver tissue sections stained with commercially available indocyanine green after intravenous injection in healthy rabbits are shown. Figure 5b Images of stained liver tissue sections from healthy rabbits after intravenous injection of the functional formulation of Example 1 are shown. Figure 5a and Figure 5b It can be seen that the liver tissues of the rabbits injected with commercially available indocyanine green and the rabbits injected with functional agents were normal, indicating that both were healthy rabbits.

[0240] In Example 1, the functional preparation was injected into the bloodstream via the marginal ear vein of rabbits with hepatic ischemia-reperfusion injury at a dose of 0.5 mg of effective fluorescent agent per kg body weight. The rabbits with hepatic ischemia-reperfusion injury were healthy New Zealand rabbits used as an injury model obtained through laparoscopic left hepatic ischemia-reperfusion surgery. The left hepatic ischemia-reperfusion surgery included the following steps: first, blunt dissection of the left hepatic Grinson's sheath was performed laparoscopically; then, sutures were passed through all blood vessels in the left hepatic region from below, and the sutures were fixed with vascular clamps, ligating all blood vessels in the left hepatic region; after 1 hour, the vascular clamps were released and the sutures were removed, allowing blood to reperfuse into the left hepatic region; the wound was sutured, and blood reperfusion was allowed for 24 hours, thus obtaining the rabbit with hepatic ischemia-reperfusion injury. These rabbits with hepatic ischemia-reperfusion injury were used to simulate patients with severe liver disease. See also... Figure 6a and Figure 6b . Figure 6a The image shows a laparoscopic fluorescence imaging of the liver and gallbladder after intravenous injection of commercially available indocyanine green in rabbits with liver ischemia-reperfusion injury. Figure 6b The image shown is a laparoscopic fluorescence imaging image of the hepatobiliary system in rabbits with liver ischemia-reperfusion injury after intravenous injection of the functional formulation of Example 1. Figure 6a As shown, one minute after injection of commercially available indocyanine green, a fluorescent signal appeared in liver area 7. From 30 to 105 minutes post-injection, the fluorescent signal in normal liver area 13 gradually decreased, indicating that the indocyanine green in normal liver area 13 was being slowly metabolized. However, the fluorescent signal in damaged liver area 14 did not decrease, indicating that the indocyanine green in damaged liver area 14 could not be metabolized normally. Therefore, it can be concluded that indocyanine green cannot be metabolized in damaged liver areas and cannot be used for biliary fluorescence imaging in patients with liver injury. Figure 6b As shown, within 5 to 60 minutes after injection of the functional formulation in Example 1, the fluorescence signal in the common bile duct was stronger than that in the liver, and the bile duct was clearly imaged with fluorescence. From 10 to 60 minutes after injection, the fluorescence signals in both the normal liver area 13 and the damaged liver area 14 decreased, indicating that the damaged liver area 14 could still metabolize the functional formulation of Example 1 normally. Therefore, the functional formulation in Example 1 can be used in patients with liver injury. See also Figure 7a and Figure 7b . Figure 7a Images of liver tissue sections stained with commercially available indocyanine green after intravenous injection in rabbits with liver ischemia-reperfusion injury are shown. Figure 7b The image shows a stained section of liver tissue from a rabbit with liver ischemia-reperfusion injury after intravenous injection of the functional formulation of Example 1. Figure 7a and Figure 7b As shown, Figure 7a and Figure 7b The capillaries around the central hepatic vein 15 were dilated, indicating that both the rabbits injected with commercially available indocyanine green and the rabbits injected with the functional preparation had liver damage.

[0241] The functional preparation in Example 1 was injected into the bloodstream via the marginal ear vein of a biliary obstruction rabbit at a dose of 0.5 mg of fluorescent agent effective content / kg body weight. The biliary obstruction rabbits were healthy New Zealand rabbits used as a model of injury obtained through laparoscopic common bile duct ligation. The common bile duct ligation procedure included the following steps: first, blunt dissection of the common bile duct was performed laparoscopically, followed by ligation of the lower end of the common bile duct, and suturing the wound. After 24 hours, the ligation was released, and bile was allowed to flow back into the duodenum, maintaining unobstructed flow at the ligation site for 4 hours to obtain the biliary obstruction rabbit. The biliary obstruction rabbits were used to simulate patients with severe biliary tract disease. See also... Figure 8a and Figure 8b . Figure 8a The image shows a laparoscopic hepatobiliary fluorescence image after intravenous injection of commercially available indocyanine green into a rabbit with bile duct obstruction. Figure 8b The image shown is a laparoscopic fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 1 into a rabbit with biliary obstruction. Figure 8a As shown, within 2 hours after intravenous injection of commercially available indocyanine green, the fluorescence signal in liver 7 did not decrease, proving that rabbits with biliary obstruction were unable to metabolize indocyanine green normally. Therefore, it can be concluded that indocyanine green cannot be used for biliary fluorescence imaging in patients with biliary tract diseases. Figure 8bAs shown, 5 minutes after intravenous injection of the functional formulation of Example 1, fluorescence signals were observed in the dilated common bile duct 12. From 10 to 60 minutes after injection, the fluorescence signal intensity of the common bile duct 12 was stronger than that of the liver 7, and the common bile duct 12 was clearly imaged with fluorescence. 10 minutes after injection, the fluorescence signal of the liver 7 rapidly decreased, and 20 minutes after injection, the fluorescence signal of the liver 7 was almost invisible. This demonstrates that the liver of the rabbit with biliary obstruction can still metabolize the functional formulation of Example 1 normally. Therefore, the functional formulation of Example 1 can be used for biliary fluorescence imaging in patients with biliary tract diseases. See also Figure 9a and Figure 9b . Figure 9a Images of liver tissue sections stained with commercially available indocyanine green after intravenous injection in rabbits with bile duct obstruction are shown. Figure 9b Images of stained liver tissue sections from rabbits with biliary obstruction after intravenous injection of the functional formulation of Example 1 are shown. Figure 9a and Figure 9b As shown, Figure 9a and Figure 9b Significant dilation of the bile ducts in the mid-liver was observed. This indicates that both the rabbits injected with commercially available indocyanine green and those injected with the functional preparation had bile duct obstruction.

[0242] The functional formulation in Example 1 was administered into the bloodstream of healthy rabbits via intravenous infusion (commonly known as intravenous drip). The infusion dose was 0.5 mg of the effective fluorescent agent content per kg of body weight, and the infusion was completed within 1 hour. See also Figure 10 , Figure 10 The image shown is a fluorescence imaging image of the liver and gallbladder of a healthy rabbit after intravenous infusion of the functional formulation of Example 1. Figure 10 As shown, 3 minutes after intravenous infusion, clear fluorescence imaging was observed in the gallbladder (6) and common bile duct (12), while no fluorescence signal was observed in the liver (7) at this time. Until the infusion was completed, no fluorescence signal was observed in the liver (7), and clear fluorescence imaging of the bile duct remained constant. This indicates that the functional formulation in Example 1 makes the dosage requirement for bile duct fluorescence imaging not significant.

[0243] The functional formulation in Example 1 was administered intravenously into the bloodstream of healthy pigs for fluorescent laparoscopic surgical navigation during cholecystectomy. The injection dose was 0.5 mg of effective fluorescent agent per kg body weight. See also Figures 11a to 11i , Figure 11a and Figure 11i Laparoscopic hepatobiliary fluorescence imaging images are shown at each stage of porcine cholecystectomy: the Greenson sheath stage, the cystic artery dissection stage, the cystic artery ligation stage, the cystic artery transection stage, the cystic duct dissection stage, the cystic duct ligation stage, the cystic duct transection stage, the gallbladder bed dissection stage, and the completion of cholecystectomy. Figures 11a to 11iAs shown, the cystic duct 15 was clearly fluorescently imaged throughout the entire operation, which enabled the cholecystectomy to proceed smoothly and avoided intraoperative damage to the cystic duct 15, which could lead to iatrogenic bile duct injury and bile fistula. Therefore, it was able to avoid serious postoperative complications and reduce the patient's suffering.

[0244] See Figures 12a to 12c , Figures 12a to 12c The comparison of cell viability after co-culturing mouse embryonic fibroblasts (NIH3T3), human umbilical vein epithelial cells (HUVECs), and normal human hepatocytes (L-O2) with different concentrations of the functional preparation from Example 1 for 24 hours is shown. The control group (0% concentration) consisted of physiological saline. All other concentrations represented the effective concentrations of the fluorescent agent. Figures 12a to 12c As shown, the effective concentration of the fluorescent agent ranged from 25 μg / mL to 200 μg / mL, and the cell survival rate exceeded 85%, demonstrating that the functional formulation of Example 1 had low cytotoxicity within the above concentration range.

[0245] See Figure 13a and Figure 13b , Figure 13a Images of stained tissue sections from various organs of live mice 14 days after injection of saline are shown. Figure 13b Images of stained tissue sections from various organs in live mice 14 days after injection of the functional formulation of Example 1 are shown. The injection dose was 2 mg of fluorescent agent effective content / kg body weight. Figure 13a and Figure 13b As shown, when the functional preparation of Example 1 was injected into the heart, liver, spleen, lungs, and kidneys, and physiological saline was injected, no significant difference was observed in any of the tissues, proving that the functional preparation of Example 1 has no significant toxicity to the internal organs of live mice.

[0246] See Figure 14 , Figure 14 Images of stained reproductive organ tissue sections from male and female mice 30 days after injection of the functional formulation of Example 1 are shown. The injection dose was 2 mg of fluorescent agent effective content / kg body weight. Figure 14 As shown, no obvious lesions were observed in the testicular and ovarian tissues, proving that the functional preparation of Example 1 has no reproductive toxicity to mice.

[0247] Example 2

[0248] The polymer in Example 2 has the molecular formula P-OEGMA. 50 -b-(LMA 20-co-(MA-IR-808)). In the polymer of Example 2, the average number of hydrophilic segments in the polymer single chain is 50, the average number of hydrophobic segments in the polymer single chain is 20, and the molar ratio of fluorescent agent to polymer single chain is 1:50. In the polymer, the average number of repeating units of polyethylene glycol is about 4.5; the first molecular weight of the polymer is 5100; the second molecular weight of the polymer is 20100. The rest of the polymer in Example 2 is the same as that in Example 1.

[0249] The preparation method of the polymer in Example 2 is the same as that in Example 1, except for the specific parameters.

[0250] In the functional formulation of Example 2, the average particle size of the polymer hydrate groups was 87.8 nanometers. The rest of the functional formulation in Example 2 was the same as that in Example 1.

[0251] The preparation method of the functional formulation in Example 2 is the same as that in Example 1.

[0252] See Figure 15a and Figure 15b , Figure 15a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 2 after intravenous injection in mice. Figure 15b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 2 into mice. The injection dosage of the functional formulation in Example 2 was the same as that in Example 1, and the injection method was tail vein injection. Figure 15a As shown, in Example 2, the functional formulation produced a fluorescent signal in the gallbladder (6) 10 minutes after injection, clear fluorescence imaging of the bile duct was achieved 30 minutes after injection, and a very weak fluorescence signal in the liver (7) was observed 45 minutes after injection. Thirty minutes after injection, the fluorescence signal of the small intestine (9) downstream of the bile duct increased. Figure 15b As shown, 5 hours after injection, gallbladder 6 was still clearly fluorescently imaged, while liver 7 showed extremely weak fluorescence signal.

[0253] Example 3

[0254] The polymer in Example 3 has the molecular formula P-OEGMA. 50 -b-(LMA 29 -co-(MA-IR-808)). In the polymer of Example 3, the average number of hydrophilic segments in the polymer single chain is 50, the average number of hydrophobic segments in the polymer single chain is 29, and the molar ratio of fluorescent agent to polymer single chain is 1:35. In the polymer, the number of repeating units of polyethylene glycol is about 4.5; the first molecular weight of the polymer is 7400; the second molecular weight of the polymer is 22400. The rest of the polymer in Example 3 is the same as that in Example 2.

[0255] The preparation method of the polymer in Example 3 is the same as that in Example 2, except for the specific parameters.

[0256] In the functional formulation of Example 3, the average particle size of the polymer hydrate groups was 104.9 nanometers. The rest of the functional formulation in Example 3 was the same as that in Example 2.

[0257] The preparation method of the functional formulation in Example 3 is the same as that in Example 2.

[0258] See Figure 16a and Figure 16b , Figure 16a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 3 after intravenous injection in mice. Figure 16b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 3 into mice. The injection dosage and method of the functional formulation of Example 3 were the same as those in Example 2. Figure 16a As shown, in Example 3, the functional formulation produced a fluorescent signal in the gallbladder (6) 10 minutes after injection, clear fluorescence imaging of the bile duct was achieved 30 minutes after injection, a very weak fluorescence signal in the liver (7) was observed 60 minutes after injection, a fluorescent signal appeared in the bladder (16) 10 minutes after injection, and a fluorescent signal appeared in the small intestine (9) 45 minutes after injection. Figure 16b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged, while the liver (7) showed almost no fluorescent signal.

[0259] Example 4

[0260] The polymer in Example 4 has the molecular formula P-OEGMA. 50 -b-(LMA7-co-(MA-IR-808)). In the polymer of Example 4, the average number of hydrophilic segments in the polymer single chain is 50, the average number of hydrophobic segments in the polymer single chain is 7, and the molar ratio of fluorescent agent to polymer single chain is 1:55. In the polymer, the average number of repeating units of polyethylene glycol is about 4.5; the first molecular weight of the polymer is 1800; the second molecular weight of the polymer is 16800. The rest of the polymer in Example 4 is the same as that in Example 2.

[0261] The preparation method of the polymer in Example 4 is the same as that in Example 2, except for the specific parameters.

[0262] In the functional formulation of Example 4, the average particle size of the polymer hydrate groups was 39.2 nanometers. The rest of the functional formulation in Example 4 was the same as that in Example 2.

[0263] The preparation method of the functional formulation in Example 4 is the same as that in Example 2.

[0264] See Figure 17a and Figure 17b , Figure 17aThe image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 4 after intravenous injection in mice. Figure 17b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 4 into mice. The injection dosage and method of the functional formulation of Example 4 were the same as those in Example 2. Figure 17a As shown, in Example 4, the functional formulation produced a fluorescent signal in the gallbladder (6) 45 minutes after injection, and clear fluorescence imaging of the bile duct was achieved 120 minutes after injection; a weak fluorescent signal appeared in the bladder (16) 5 minutes after injection; a fluorescent signal appeared in the small intestine (9) 30 minutes after injection; and a fluorescent signal appeared in the colon (10) 180 minutes after injection. Figure 17b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, while the liver 7 showed almost no fluorescence signal.

[0265] Example 5

[0266] The polymer in Example 5 has the molecular formula P-OEGMA. 81 -b-(LMA 11 -co-(MA-IR-808)). In the polymer of Example 5, the average number of hydrophilic segments in the polymer single chain is 81, the average number of hydrophobic segments in the polymer single chain is 11, and the molar ratio of fluorescent agent to polymer single chain is 1:55. In the polymer, the average number of repeating units of polyethylene glycol is about 4.5; the first molecular weight of the polymer is 2800; the second molecular weight of the polymer is 27100. The rest of the polymer in Example 5 is the same as that in Example 2.

[0267] The preparation method of the polymer in Example 5 is the same as that in Example 2, except for the specific parameters.

[0268] In the functional formulation of Example 5, the average particle size of the polymer hydrate groups was 394.7 nanometers. The rest of the functional formulation in Example 5 was the same as that in Example 2.

[0269] The preparation method of the functional formulation in Example 5 is the same as that in Example 2.

[0270] See Figure 18a and Figure 18b , Figure 18a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 5 after intravenous injection in mice. Figure 18b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 5 into mice. The injection dosage and method of the functional formulation of Example 5 were the same as those in Example 2. Figure 18a As shown, in Example 5, the functional formulation produced a fluorescent signal in the gallbladder (6) 30 minutes after injection, and the bile duct was clearly imaged with fluorescence 30 minutes after injection; a fluorescent signal appeared in the small intestine (9) 15 minutes after injection. Figure 18b As shown, 3 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) showed fluorescence signals.

[0271] Example 6

[0272] The polymer in Example 6 has the molecular formula P-OEGMA. 50 -b-(EMA 28 -co-(MA-IR-808)). In the polymer of Example Six, the hydrophobic group of the hydrophobic linker is ethyl. The average number of hydrophilic links in the polymer single chain is 50, the average number of hydrophobic links in the polymer single chain is 28, and the molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the average number of repeating units of polyethylene glycol is about 4.5; the first molecular weight of the polymer is 3200; the second molecular weight of the polymer is 18200.

[0273] The rest of the polymer in Example 6 is the same as in Example 2.

[0274] In the polymer preparation method of Example 6, the hydrophobic monomer used is ethyl methacrylate (EMA), whose hydrophobic group is ethyl. The rest of the polymer preparation method is similar to that of Example 2.

[0275] In the functional formulation of Example 6, the average particle size of the polymer hydrate groups was 30.7 nanometers. The rest of the functional formulation in Example 6 was the same as that in Example 2.

[0276] The preparation method of the functional formulation in Example 6 is the same as that in Example 2.

[0277] See Figure 19a and Figure 19b , Figure 19a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 6 after intravenous injection in mice. Figure 19b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 6 into mice. The injection dosage and injection method of the functional formulation of Example 6 were the same as those in Example 2. Figure 19a As shown, in Example 6, the functional formulation produced a fluorescent signal in the gallbladder (6) 30 minutes after injection, and the bile duct was clearly imaged with fluorescence; a weak fluorescent signal appeared in the bladder (16) 15 minutes after injection; and a fluorescent signal appeared in the small intestine (9) 30 minutes after injection. Figure 19b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, while the liver 7 showed almost no fluorescence signal.

[0278] Example 7

[0279] The polymer in Example 7 has the molecular formula P-OEGMA.50 -b-(HMA 45 -co-(MA-IR-808)). In the polymer of Example 7, the hydrophobic group of the hydrophobic linker is hexyl. The average number of hydrophilic links in the polymer single chain is 50, the average number of hydrophobic links in the polymer single chain is 45, and the molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the average number of repeating units of polyethylene glycol is about 4.5; the first molecular weight of the polymer is 7700; the second molecular weight of the polymer is 22700.

[0280] The rest of the polymer in Example 7 is the same as in Example 2.

[0281] In the preparation method of the polymer in Example 7, the hydrophobic monomer used is hexyl methacrylate (HMA), whose hydrophobic group is hexyl. The rest of the preparation method of the polymer is similar to that in Example 2.

[0282] In the functional formulation of Example 7, the average particle size of the polymer hydrate groups was 112.6 nanometers. The rest of the functional formulation in Example 7 was the same as that in Example 2.

[0283] The preparation method of the functional formulation in Example 7 is the same as that in Example 2.

[0284] See Figure 20a and Figure 20b , Figure 20a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 7 after intravenous injection in mice. Figure 20b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 7 into mice. The injection dosage and method of the functional formulation of Example 7 were the same as those in Example 2. Figure 20a As shown, in Example 7, the functional formulation produced a fluorescent signal in the gallbladder (6) 3 minutes after injection, and clear fluorescence imaging of the bile duct was achieved 15 minutes after injection; a fluorescent signal appeared in the bladder (16) 10 minutes after injection; a fluorescent signal appeared in the duodenum (8) 10 minutes after injection; a fluorescent signal appeared in the small intestine (9) 15 minutes after injection; and a fluorescent signal appeared in the colon (10) 120 minutes after injection. Figure 20b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0285] Example 8

[0286] The polymer in Example 8 has the molecular formula P-OEGMA. 50 -b-(cHMA 46-co-(MA-IR-808)). In the polymer of Example 8, the hydrophobic group of the hydrophobic linker is cyclohexyl. The average number of hydrophilic links in the polymer single chain is 50, the average number of hydrophobic links in the polymer single chain is 46, and the molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the average number of repeating units of polyethylene glycol is about 4.5; the first molecular weight of the polymer is 7800; the second molecular weight of the polymer is 22800. The rest of the polymer in Example 8 is the same as that in Example 2.

[0287] In the preparation method of the polymer in Example 8, the hydrophobic monomer used is cyclohexyl methacrylate (cHMA), and the hydrophobic group is cyclohexyl. The rest of the preparation method of the polymer is similar to that in Example 2.

[0288] In the functional formulation of Example 8, the average particle size of the polymer hydrate groups was 99.1 nanometers. The rest of the functional formulation in Example 8 was the same as that in Example 2.

[0289] The preparation method of the functional formulation in Example 8 is the same as that in Example 2.

[0290] See Figure 21a and Figure 21b , Figure 21a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 8 after intravenous injection in mice. Figure 21b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 8 into mice. The injection dosage and method of the functional formulation of Example 8 were the same as those in Example 2. Figure 21a As shown, in Example 8, the functional formulation produced a fluorescent signal in the gallbladder (6) 15 minutes after injection, and the bile duct was clearly imaged with fluorescence; a fluorescent signal appeared in the bladder (16) 5 minutes after injection; a fluorescent signal appeared in the duodenum (8) 15 minutes after injection; a fluorescent signal appeared in the small intestine (9) 30 minutes after injection; and a fluorescent signal appeared in the colon (10) 120 minutes after injection. Figure 21b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0291] Example 9

[0292] The polymer in Example 9 has the molecular formula P-OEGMA. 50 -b-(SMA 10-co-(MA-IR-808)). In the polymer of Example 9, the hydrophobic group of the hydrophobic linker is octadecyl. The average number of hydrophilic links in the polymer single chain is 50, the average number of hydrophobic links in the polymer single chain is 10, and the molar ratio of fluorescent agent to polymer single chain is 1:55. In the polymer, the average number of repeating units of polyethylene glycol is about 4.5; the first molecular weight of the polymer is 3400; the second molecular weight of the polymer is 18400. The rest of the polymer in Example 9 is the same as that in Example 2.

[0293] In the preparation method of the polymer in Example 9, the hydrophobic monomer used is octadecyl methacrylate (SMA), and the hydrophobic group is octadecyl. The rest of the preparation method of the polymer is similar to that in Example 2.

[0294] In the functional formulation of Example 9, the average particle size of the polymer hydrate groups was 36.1 nanometers. The rest of the functional formulation in Example 9 was the same as that in Example 2.

[0295] The preparation method of the functional formulation in Example 9 is the same as that in Example 2.

[0296] See Figure 22a and Figure 22b , Figure 22a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 9 after intravenous injection in mice. Figure 22b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 9 into mice. The injection dosage and method of the functional formulation of Example 9 are the same as those in Example 2. Figure 22a As shown, in Example 9, the functional formulation produced a fluorescent signal in the gallbladder (6) 30 minutes after injection, and clear fluorescence imaging of the bile duct was achieved 60 minutes after injection; a fluorescent signal appeared in the bladder (16) 5 minutes after injection; a fluorescent signal appeared in the small intestine (9) 10 minutes after injection; and a fluorescent signal appeared in the colon (10) 120 minutes after injection. Figure 22b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0297] Example 10

[0298] The polymer in Example 10 has the molecular formula P-OEGMA. 50 -b-(nBMA 30-co-(MA-IR-808)). In the polymer of Example 10, the hydrophobic group of the hydrophobic linkage is n-butyl. The average number of hydrophilic linkages in the polymer single chain is 50, the average number of hydrophobic linkages in the polymer single chain is 30, and the molar ratio of fluorescent agent to polymer single chain is 1:40. In the polymer, the average number of repeating units of polyethylene glycol is about 4.5; the first molecular weight of the polymer is 4300; the sum of the second molecular weights of the polymer is 19300. The rest of the polymer in Example 10 is the same as that in Example 2.

[0299] In the preparation method of the polymer in Example 10, the hydrophobic monomer used is n-butyl methacrylate (nBMA), and the hydrophobic group is n-butyl. The rest of the preparation method of the polymer is similar to that in Example 2.

[0300] In the functional formulation of Example 10, the average particle size of the polymer hydrate groups was 61.3 nanometers. The rest of the functional formulation in Example 10 was the same as that in Example 2.

[0301] The preparation method of the functional formulation in Example 10 is the same as that in Example 2.

[0302] See Figure 23a and Figure 23b , Figure 23a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 10 after intravenous injection in mice. Figure 23b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 10 into mice. The injection dosage and method of the functional formulation of Example 10 are the same as those in Example 2. Figure 23a As shown, in Example 10, a fluorescent signal appeared in the bladder 16 5 minutes after injection of the functional formulation; a fluorescent signal appeared in the colon 10 120 minutes after injection. Figure 23b As shown, 3 hours after injection, the gallbladder 6 was still clearly fluorescently imaged with a strong fluorescence signal, the liver 7 had almost no fluorescence signal, and the colon 10 still had a fluorescence signal.

[0303] Example 11

[0304] The polymer in Example 11 has the molecular formula P-OEGMA. 50 -b-(EMA 45 -co-(MA-IR-808)). In the polymer of Example 11, the hydrophobic group of the hydrophobic linker is methyl. The average number of hydrophilic links in the polymer single chain is 50, the average number of hydrophobic links in the polymer single chain is 45, and the molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the number of repeating units of polyethylene glycol is about 4.5, the first molecular weight of the polymer is 4600, and the second molecular weight of the polymer is 19600.

[0305] The rest of the polymer in Example 11 is the same as in Example 2.

[0306] In the preparation method of the polymer in Example 11, the hydrophobic monomer used is methyl methacrylate (EMA), and the rest of the preparation method of the polymer is similar to that in Example 2.

[0307] In the functional formulation of Example 11, the average particle size of the polymer hydrate groups was 67.2 nanometers. The rest of the functional formulation in Example 11 was the same as that in Example 2.

[0308] The preparation method of the functional formulation in Example 11 is the same as that in Example 2.

[0309] See Figure 24a and Figure 24b , Figure 24a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 11 after intravenous injection in mice. Figure 24b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 11 into mice. The injection dosage and method of the functional formulation of Example 11 are the same as those in Example 2. Figure 24a As shown, the functional formulation of Example 11 showed a fluorescent signal in the gallbladder (6) 10 minutes after injection, and clear fluorescence imaging of the bile duct was achieved 30 minutes after injection; a fluorescent signal appeared in the duodenum (8) 15 minutes after injection, and a fluorescent signal appeared in the colon (10) 120 minutes after injection. Figure 24b As shown, 4 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0310] Example 12

[0311] The polymer in Example 12 has the molecular formula P-PEGMA. 50 -b-(LMA 21 -co-(MA-IR-808)). PEGMA represents a hydrophilic monomer with an average number of 6-8 repeating units in polyethylene glycol (the product is commercially available, and the nominal average number of repeating units in polyethylene glycol is 6-8, specifically approximately 6.2 as measured). Specifically, it is polyethylene glycol methacrylate. In the polymer of Example Twelve, the hydrophobic group of the hydrophobic chain segment is dodecyl. The average number of hydrophilic chains in the polymer single chain is 50, the average number of hydrophobic chains in the polymer single chain is 21, and the molar ratio of fluorescent agent to polymer single chain is 1:45. In the polymer, the average number of repeating units in polyethylene glycol is 6.2; the first molecular weight of the polymer is 5400; and the sum of the second molecular weights of the polymer is 23400. The remaining parts of the polymer in Example Twelve are the same as in Example Two.

[0312] In the polymer preparation method of Example Twelve, the hydrophilic monomer is PEGMA, specifically polyethylene glycol methacrylate. The rest of the polymer preparation method is similar to that of Example Two.

[0313] In the functional formulation of Example Twelve, the average particle size of the polymer hydrate groups was 68.9 nanometers. The rest of the functional formulation in Example Twelve was the same as that in Example Two.

[0314] The preparation method of the functional formulation in Example Twelve is the same as that in Example Two.

[0315] See Figure 25a and Figure 25b , Figure 25a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example Twelve after intravenous injection in mice; Figure 25b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example Twelve into mice. The injection dosage and method of the functional formulation of Example Twelve are the same as those in Example Two. Figure 25a As shown, in Example 12, the functional formulation produced a fluorescent signal in the gallbladder (6) 15 minutes after injection, and clear fluorescence imaging of the bile duct was achieved 60 minutes after injection; fluorescent signals appeared in the duodenum (8) and bladder (16) 10 minutes after injection; fluorescent signals appeared in the small intestine (9) 60 minutes after injection; and fluorescent signals appeared in the colon (10) 180 minutes after injection. Figure 25b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0316] Example 13

[0317] In Example 13, the polymer single chain includes a hydrophilic segment and a hydrophobic segment of block copolymerization. The hydrophilic segment includes multiple hydrophilic links, the polymeric group of which is methacrylate, and the average number of repeating units in the polyethylene glycol is approximately 4.5. The hydrophobic segment includes multiple hydrophobic links, the polymeric group of which is methacrylate, and the hydrophobic group is dodecyl. For the polymer, at least a portion of the polymer single chain includes functional links embedded in the hydrophilic segment, and at least a portion of the polymer single chain includes links to be modified embedded in the hydrophilic segment. Each functional link is conjugated with a fluorescent agent, the molar ratio of the fluorescent agent to the polymer single chain is 1:35, and the polymeric group of the functional link is acrylate. The link to be modified is conjugated with a first reactive group, which is used to conjugate the fluorescent agent through a substitution reaction or coupling reaction, the molar ratio of the first reactive group to the polymer single chain is 1:1.35, and the polymeric group of the link to be modified is acrylate. In this example, the first reactive group is succinimide ester. In other embodiments, the first reactive group may also be any one or more of succinimide ester, maleimide ester, azide, alkynyl, amino, mercapto, hydroxyl and aldehyde groups.

[0318] In Example Thirteen, the polymer molecular formula is P-(OEGMA) 48 -co-(A-IR-808))-b-LMA 16 In the polymer, the average number of hydrophilic segments in the hydrophilic chain is 48, and the average number of repeating polyethylene glycol units in the hydrophilic segments is approximately 4.5; the average number of hydrophobic segments in the hydrophobic chain is 16, and the hydrophobic group in the hydrophobic segments is dodecyl. On average, there is one functional segment in every 35 polymer chains. The fluorescent agent conjugated to the functional segment is IR-808, a derivative of IR-783. In Example Thirteen, the first molecular weight of the polymer is 4100, and the second molecular weight of the polymer is 18500.

[0319] The polymer of Example Thirteen is otherwise identical to that of Example Two.

[0320] The preparation method of the polymer in Example Thirteen is as follows:

[0321] Step 1: Prepare hydrophilic segments with pre-modified chain segments via living radical polymerization:

[0322] In Example 13, the living radical polymerization reaction used was a reversible addition-fragmentation chain transfer (RAFT) polymerization reaction. The chain transfer agent was 4-cyano-4-(thiobenzoyl)valerate (CTA). The hydrophilic monomer was oligomeric polyethylene glycol methacrylate (OEGMA, with an average number of repeating units of approximately 4.5 and a polymeric group of methacrylate). The monomer to be modified was N-acryloyloxysuccinimide (NAS), wherein the polymeric group was acrylate and the first reactive group was succinimide ester. The initiator was azobisisobutyronitrile (AIBN). The solvent for calibration by monitoring 1H NMR spectroscopy was anisole.

[0323] First, CTA 56 mg (0.2 mmol), OEGMA 3.15 g (10.5 mmol), NAS 34 mg (0.2 mmol), and AIBN 3.3 mg (0.02 mmol) were dissolved in 15 mL of 1,4-dioxane, and 3 to 4 drops of anisole were added. The mixture was reacted in a Shrek tube, and the conversion rate was monitored by 1H NMR spectroscopy. The reaction conditions were: after deoxygenation, the temperature was raised to 70 °C. During the reaction, an appropriate amount of the mixture was taken and the conversion rate was monitored by 1H NMR spectroscopy. When the conversion rate reached 91.5%, oxygen was introduced to stop the reaction. After the reaction was completed, the mixture was cooled to room temperature, and the supernatant was removed by precipitation in n-hexane. After vacuum drying at room temperature, 2.87 g of hydrophilic segment was obtained. The molecular formula of the hydrophilic segment with the prepared modified segment is P-OEGMA. 48 -co-NAS. The hydrophilic monomer forms a hydrophilic chain segment in the hydrophilic chain segment, and the monomer to be modified is inserted into the hydrophilic chain segment. After the monomer to be modified is inserted, a chain segment to be modified is formed.

[0324] Step 2: Block polymerization of hydrophobic segments via living radical polymerization

[0325] The chain transfer agent is the hydrophilic segment (P-OEGMA) prepared in step 1. 48 The hydrophobic monomer is lauryl methacrylate (LMA, with a dodecyl hydrophobic group and a methacrylate polymeric group), the initiator is AIBN, and the calibration solvent for 1H NMR spectroscopy monitoring is anisole.

[0326] First, P-OEGMA 481.45 g (0.1 mmol) of -co-NAS, 458 mg (1.8 mmol) of LMA, and 3.3 mg of AIBN were dissolved in 15 mL of N,N-dimethylformamide (DMF), and 3 to 4 drops of anisole were added. The mixture was reacted in a Shrek tube, and the conversion was monitored by 1H NMR spectroscopy. The reaction conditions were: deoxygenation followed by heating to 70 °C. During the reaction, a suitable amount of the mixture was taken and the conversion was monitored by 1H NMR spectroscopy. The reaction was stopped when oxygen was introduced when the conversion reached 88.9%. After the reaction, the mixture was cooled to room temperature, the DMF was evaporated to dryness, and then redissolved in 15 mL of tetrahydrofuran. The supernatant was then removed by precipitation in n-hexane, and the mixture was dried under vacuum at room temperature to obtain 1.67 g of intermediate polymer. The molecular formula of the intermediate polymer is P-(OEGMA). 48 -co-NAS)-b-LMA 16 In this process, hydrophobic monomers form hydrophobic links within the hydrophobic chain segments.

[0327] Step 3: Modify the intermediate polymer with fluorescent agent via substitution reaction

[0328] The intermediate polymer P-(OEGMA) participated in the substitution reaction. 48 -co-NAS)-b-LMA 16 The second unit is IR-808-NH2, which is formed by the connection of a fluorescent agent and a second reactive group. The fluorescent agent is IR-808, a derivative of IR-783, and the second reactive group is an amino group.

[0329] 0.9g P-(OEGMA) 48 -co-NAS)-b-LMA 16 0.05 mmol of IR-808-NH2 and 5 mg of IR-808-NH2 (0.005 mmol) were dissolved in 10 mL of DMF, and 1 mL of triethylamine was added to create an alkaline environment. The mixture was stirred at room temperature in the dark for 24 hours. After the reaction was complete, 2 mL of deionized water was added to the mixture, and it was placed in a 5 kDa dialysis bag and dialyzed against deionized water for 48 hours to remove unreacted IR-808-NH2. After complete drying, 0.87 g of P-(OEGMA) was obtained. 50 -co-(A-IR-808))-b-LMA 16In this process, the substitution reaction between the first and second reactive groups on the intermediate polymer is a probabilistic substitution reaction with a reaction efficiency of 28.6%. This allows the fluorescent agent to be probabilistically modified into functional segments through substitution, while the unmodified fluorescent agent segments become the segments to be modified. In other embodiments, for example, if the first reactive group is an aldehyde and the second reactive group is an amine, modification can be achieved through a coupling reaction without the need for an alkaline environment. In this case, the coupling reaction can also occur simultaneously with the polymerization reaction. However, due to the higher polymerization temperature, some fluorescent agents will inevitably become inactive.

[0330] In Example 13, the average number of repeating polyethylene glycol units in the polymer is controlled by controlling the number of repeating units in the hydrophilic monomer. The first molecular weight of the polymer is controlled by controlling the degree of polymerization of the hydrophobic monomer. The second molecular weight of the polymer is controlled by controlling the degree of polymerization of both the hydrophobic and hydrophilic monomers. The degree of polymerization of the hydrophobic monomer is controlled by controlling the molar ratio of the hydrophobic monomer to the chain transfer agent and the conversion rate of the living radical polymerization reaction used to polymerize the hydrophobic segments. The degree of polymerization of the hydrophilic monomer is also controlled by controlling the molar ratio of the hydrophilic monomer to the chain transfer agent and the conversion rate of the living radical polymerization reaction used to polymerize the hydrophilic segments. In other examples, when atom transfer radical polymerization (ATRP) is used, the degree of polymerization of the hydrophobic monomer is controlled by controlling the molar ratio of the hydrophobic monomer to the initiator and the conversion rate of the living radical polymerization reaction used to polymerize the hydrophobic segments, and the degree of polymerization of the hydrophilic monomer is controlled by controlling the molar ratio of the hydrophilic monomer to the initiator and the conversion rate of the living radical polymerization reaction used to polymerize the hydrophilic segments.

[0331] In the functional formulation of Example Thirteen, the average particle size of the polymer hydrate groups was 56.3 nanometers. The rest of the functional formulation in Example Thirteen was the same as that in Example Two.

[0332] The preparation method of the functional formulation in Example 13 is the same as that in Example 2.

[0333] See Figure 26a and Figure 26b , Figure 26a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example Thirteen after intravenous injection in mice. Figure 26b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example Thirteen into mice. The injection dosage and method of the functional formulation of Example Thirteen are the same as those in Example Two. Figure 26a As shown, in Example 13, the functional formulation produced a fluorescent signal in the gallbladder (6) 10 minutes after injection, and the bile duct was clearly imaged with fluorescence; a fluorescent signal appeared in the small intestine (9) 30 minutes after injection; and a fluorescent signal appeared in the colon (10) 120 minutes after injection. Figure 26bAs shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0334] Example 14

[0335] In Example 14, the polymer single chain has a linear topological structure. The polymer single chain sequentially includes a first hydrophilic segment, a hydrophobic segment, and a second hydrophilic segment, all of which are hydrophilic. Each hydrophilic segment has a brush-like structure and includes multiple hydrophilic links, which are conjugated with polyethylene glycol. The hydrophobic segment includes multiple hydrophobic links, which are conjugated with hydrophobic groups. At least a portion of the polymer single chain also includes functional links embedded in the hydrophobic segments. At least a portion of the functional links are conjugated with a fluorescent agent, with a molar ratio of functional links to polymer single chains of 1:24 and a molar ratio of fluorescent agent to polymer single chains of 1:40.

[0336] In Example Fourteen, the polymer molecular formula is P-OEGMA 24 -b-(LMA 20 -co-(MA-IR-808))-b-OEGMA 25 In Example 14, the polymer groups used for the hydrophilic monomer, hydrophobic monomer, and functional monomer were all methacrylate groups. In the polymer, the average number of hydrophilic segments in the hydrophilic chain was 49, and the average number of repeating polyethylene glycol units in the hydrophilic segments was approximately 4.5. The average number of hydrophobic segments in the hydrophobic chain was 20, and the hydrophobic group in the hydrophobic segments was dodecyl. On average, there was one functional segment in every 24 polymer chains. The fluorescent agent conjugated to the functional segment was IR-783, and the substance formed after the fluorescent agent conjugated to the functional segment deactivated was the substance formed after the deactivation of IR-783 derivative IR-808. In Example 14, the first molecular weight of the polymer was 5100, and the second molecular weight of the polymer was 19800.

[0337] The polymer in Example 14 is otherwise identical to the polymer in Example 2.

[0338] The polymer preparation method in Example Fourteen comprises three steps. Steps 1 and 2 are similar to steps 1 and 2 in Example One, respectively. Step 3 essentially involves using the intermediate polymer obtained in step 2 as a chain transfer agent to polymerize again in the manner described in step 1 to form hydrophilic segments.

[0339] In the functional formulation of Example Fourteen, the average particle size of the polymer hydrate groups was 79.3 nanometers. The rest of the functional formulation in Example Fourteen was the same as that in Example Two.

[0340] The preparation method of the functional formulation in Example 14 is the same as that in Example 2.

[0341] See Figure 27a and Figure 27b , Figure 27a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example Fourteen after intravenous injection in mice. Figure 27b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example Fourteen into mice. The injection dosage and method of the functional formulation of Example Fourteen are the same as those in Example Two. Figure 27a As shown, in Example 14, the functional formulation produced a fluorescent signal in the gallbladder (6) 10 minutes after injection, and the bile duct was clearly imaged with fluorescence; a fluorescent signal appeared in the duodenum (8) 10 minutes after injection; a fluorescent signal appeared in the small intestine (9) 30 minutes after injection; and a fluorescent signal appeared in the colon (10) 120 minutes after injection. Figure 27b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0342] Example 15

[0343] In Example 15, the polymer single chain has a linear topological structure. The polymer single chain sequentially includes a first hydrophobic segment, a hydrophilic segment, and a second hydrophobic segment, all of which are block copolymers. The first and second hydrophobic segments are both hydrophobic. The hydrophilic segment has a brush-like structure and includes multiple hydrophilic segments, each conjugated with polyethylene glycol. The hydrophobic segment includes multiple hydrophobic segments, each conjugated with a hydrophobic group. At least a portion of the polymer single chain also includes functional segments embedded in the hydrophilic segments. At least a portion of the functional segments are conjugated with a fluorescent agent, with a molar ratio of functional segments to polymer single chains of 1:15 and a molar ratio of fluorescent agent to polymer single chains of 1:25.

[0344] In Example 15, the polymer molecular formula is P-LMA 11 -b-(OEGMA 51 -co-(MA-IR-808))-b-LMA 12 In Example 15, the polymer groups used for the hydrophilic monomer, hydrophobic monomer, and functional monomer were all methacrylate groups. In the polymer, the average number of hydrophilic segments in the hydrophilic chain was 51, and the average number of repeating polyethylene glycol units in the hydrophilic segments was approximately 4.5. The average number of hydrophobic segments in the hydrophobic chain was 23, and the hydrophobic group in the hydrophobic segments was dodecyl. On average, there was one functional segment in every 15 polymer chains. The fluorescent agent conjugated to the functional segment was IR-808, a derivative of IR-783, and the substance formed after the fluorescent agent conjugated to the functional segment deactivated was the substance formed after the deactivation of IR-808, a derivative of IR-783. In Example 15, the first molecular weight of the polymer was 5900, and the second molecular weight of the polymer was 21200.

[0345] The polymer of Example 15 is otherwise identical to that of Example 2.

[0346] The preparation method of the polymer in Example 15 is as follows:

[0347] Step 1: Preparation of the first hydrophobic segment via living radical polymerization

[0348] In Example 15, the living radical polymerization reaction used was a reversible addition-fragmentation chain transfer (RAFT) polymerization reaction. The chain transfer agent was 4-cyano-4-(thiobenzoyl)valerate (CTA). The hydrophobic monomer was lauryl methacrylate (LMA). The initiator was azobisisobutyronitrile (AIBN). The solvent for calibration by monitoring 1H NMR spectroscopy was anisole.

[0349] First, CTA 56 mg (0.2 mmol), LMA 610 mg (2.4 mmol), and AIBN 3.3 mg (0.02 mmol) were dissolved in 15 mL of 1,4-dioxane, and 3 to 4 drops of anisole were added. The mixture was reacted in a Shrek tube, and the conversion rate was monitored by 1H NMR spectroscopy. The reaction conditions were: after deoxygenation, the temperature was raised to 70 °C. During the reaction, an appropriate amount of the mixture was taken and the conversion rate was monitored by 1H NMR spectroscopy. When the conversion rate reached 91.8%, oxygen was introduced to stop the reaction. After the reaction was completed, the mixture was cooled to room temperature, and the supernatant was removed by precipitation in ethanol. After vacuum drying at room temperature, 0.54 g of the first hydrophobic segment was obtained. The molecular formula of the first hydrophobic segment is P-LMA. 11 Hydrophobic monomers form hydrophobic links in the first hydrophobic chain segment.

[0350] Step 2: Polymerize hydrophilic segments via living radical polymerization.

[0351] The chain transfer agent is P-LMA 11 The hydrophilic monomer is oligomeric polyethylene glycol methacrylate (OEGMA), specifically polyethylene glycol methyl ether methacrylate, with an average of approximately 4.5 repeating polyethylene glycol units. The functional monomer is MA-IR-808. The initiator is azobisisobutyronitrile (AIBN). The calibration solvent for 1H NMR spectroscopy monitoring is anisole.

[0352] First, P-LMA 110.28 g (0.1 mmol), 1.68 g (5.6 mmol) of OEGMA, 10 mg of MA-IR-808, and 3.3 mg of AIBN were dissolved in 15 mL of N,N-dimethylformamide (DMF), and 3 to 4 drops of anisole were added. The mixture was reacted in a Shrek tube, and the conversion was monitored by 1H NMR spectroscopy. The reaction conditions were: after deoxygenation, the temperature was raised to 70 °C. During the reaction, an appropriate amount of the mixture was taken and the conversion was monitored by 1H NMR spectroscopy. When the conversion reached 90.9%, oxygen was introduced to stop the reaction. After the reaction, the mixture was cooled to room temperature, the DMF was evaporated to dryness, and redissolved in 15 mL of tetrahydrofuran. The supernatant was then removed by precipitation in n-hexane, and the mixture was dried under vacuum at room temperature to obtain 1.67 g of polymer. The molecular formula of the intermediate polymer is P-LMA. 11 -b-(OEGMA 51 -co-(MA-IR-808)). In this reaction, hydrophilic monomers form hydrophilic repeating segments within the hydrophilic chain; functional monomers form functional repeating segments within the hydrophilic chain. During the reaction, due to the high temperature of the polymerization reaction, some of the fluorescent agent becomes inactive. The molar ratio of the substance formed after the fluorescent agent becomes inactive to the fluorescent agent is approximately 4:6.

[0353] Step 3: Preparation of polymers via living free radical reaction

[0354] The chain transfer agent is the intermediate polymer P-LMA prepared in step 2. 11 -b-(OEGMA 51 -co-(MA-IR-808)), the hydrophobic monomer is lauryl methacrylate (LMA, the hydrophobic group is dodecyl, the polymeric group is methacrylate), the initiator is AIBN, and the calibration solvent for 1H NMR spectroscopy monitoring is anisole.

[0355] First, P-LMA 11 -b-(OEGMA 51 0.9 g (0.05 mmol) of 1,4-dioxane (-co-(MA-IR-808)), 180 mg (0.7 mmol) of LMA, and 3.3 mg of AIBN were dissolved in 15 mL of 1,4-dioxane, and 3 to 4 drops of anisole were added. The mixture was reacted in a Shrek tube, and the conversion rate was monitored by 1H NMR spectroscopy. The reaction conditions were: after deoxygenation, the temperature was raised to 70 °C. During the reaction, an appropriate amount of the mixture was taken and the conversion rate was monitored by 1H NMR spectroscopy. When the conversion rate reached 86.1%, oxygen was introduced to stop the reaction. After the reaction was completed, the temperature was lowered to room temperature, and the supernatant was removed by precipitation in n-hexane. After removing the supernatant, the polymer was dried under vacuum at room temperature to obtain 1.0 g of polymer. The molecular formula of the polymer is P-LMA. 11 -b-(OEGMA 51 -co-(MA-IR-808))-b-LMA 12In this process, hydrophobic monomers form hydrophobic links within the hydrophobic chain segments.

[0356] In the functional formulation of Example 15, the average particle size of the polymer hydrate groups was 96.7 nanometers. The rest of the functional formulation in Example 15 was the same as that in Example 2.

[0357] The preparation method of the functional formulation in Example 15 is the same as that in Example 2.

[0358] See Figure 28a and Figure 28b , Figure 28a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 15 after intravenous injection in mice. Figure 28b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 15 into mice. The injection dosage and method of the functional formulation of Example 15 were the same as those in Example 2. Figure 28a As shown, in Example 15, the functional formulation produced a fluorescent signal in the gallbladder (6) 30 minutes after injection, and the bile duct was clearly imaged with fluorescence; a fluorescent signal appeared in the bladder 3 minutes after injection; a fluorescent signal appeared in the duodenum (8) 10 minutes after injection; a fluorescent signal appeared in the small intestine (9) 15 minutes after injection; and a fluorescent signal appeared in the colon (10) 180 minutes after injection. Figure 28b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0359] Example 16

[0360] In Example 16, the polymer single chain exhibits a star-shaped topological structure. The polymer single chain comprises six hydrophilic segments and six hydrophobic segments, all block copolymerized. The star structure has six arms, each arm containing one hydrophobic segment and one hydrophilic segment. On each arm, the hydrophobic segment is closer to the center of the star than the hydrophilic segment. The hydrophilic segments have a brush-like structure and include multiple hydrophilic links, each conjugated with polyethylene glycol. The hydrophobic segments also include multiple hydrophobic links, each conjugated with a hydrophobic group. At least a portion of the polymer single chain contains a functional link, which is embedded within any one or more hydrophobic segments. At least a portion of the polymer single chain includes a link to be modified, which is conjugated with a first reactive group, namely succinimidyl ester. The link to be modified is embedded within any one or more hydrophobic segments. The molar ratio of the functional link to the polymer single chain is 1:30, and the molar ratio of the fluorescent agent 5 to the polymer single chain is also 1:30. The molar ratio of the modified repeating unit to the polymer single chain is 1 to 1.25.

[0361] In Example Sixteen, the polymer molecular formula is 6-Arm-P-(LMA) 20 -co-(A-IR-808))-b-OEGMA51 In the polymer, the average number of hydrophilic segments is 51, and the number of repeating polyethylene glycol units in each hydrophilic segment is approximately 4.5. The average number of hydrophobic segments is 20, and the hydrophobic group in each hydrophobic segment is dodecyl. On average, there is one functional segment in every 30 polymer chains. The fluorescent agent conjugated to the functional segment is IR-808, a derivative of IR-783. In Example Sixteen, the first molecular weight of the polymer is 3600, and the second molecular weight of the polymer is 21600.

[0362] The other parts of the polymer in Example 16 are the same as those in Example 1.

[0363] The preparation method of the polymer in Example Sixteen is as follows:

[0364] Step 1: Preparation of a six-armed initiator

[0365] 2.54 g (10 mmol) of dipentaerythritol was dispersed in 50 mL of dichloromethane, and 15 mL of triethylamine was added. The mixture was stirred in an ice bath for 30 minutes. Then, 10 mL of 2-bromoisobutyryl bromide was added through a dropping funnel, while maintaining stirring in an ice bath. The reaction was then carried out at room temperature for 24 hours. After the reaction was complete, an appropriate amount of deionized water was added dropwise to quench unreacted 2-bromoisobutyryl bromide. The mixture was filtered to remove insoluble matter, and the organic phase was further extracted with deionized water to remove water-soluble impurities. After the organic phase was completely evaporated to dryness, column chromatography (using ethyl acetate and n-hexane as eluents, with a volume ratio of 1:1) was used to obtain the 6-arm initiator 6-Arm-Initiator, whose structural formula is:

[0366]

[0367] Step 2: Prepare hydrophobic segments with the pre-modified chain segments via active free radical reaction.

[0368] In Example 16, the living radical polymerization reaction was atom transfer radical polymerization (ATRP). The initiator was 6-Arm-Initiator. The hydrophobic monomer was LMA. The monomer to be modified was NAS. The catalyst was copper bromide (CuBr2). The copper ion ligand was tris(2-dimethylaminoethyl)amine (me6tren). The solvent for NMR spectroscopy monitoring and calibration was anisole.

[0369] First, 114.8 mg (0.1 mmol) of 6-Arm-Initiator, 17 mg (0.1 mmol) of NAS, 560 mg (2.2 mmol) of LMA, and 5.4 mg (0.024 mmol) of CuBr2 were dissolved in 5 mL of DMF, and 3 to 4 drops of anisole were added. After deoxygenation, 28 mg (0.12 mmol) of me6tren was injected. The mixture was reacted in a Shrek tube, and the conversion rate was monitored by 1H NMR spectroscopy. The reaction conditions were room temperature. During the reaction, an appropriate amount of the mixture was taken and the conversion rate was monitored by 1H NMR spectroscopy. When the conversion rate reached 91.3%, oxygen was introduced to stop the reaction. After the reaction, the DMF was evaporated, and then redissolved in tetrahydrofuran and CuBr2 was removed by passing through a neutral alumina column. The mixture was precipitated in n-hexane and dried under vacuum at room temperature for 24 hours to obtain 0.61 g of intermediate polymer. The molecular formula of the intermediate polymer is 6-Arm-P-(LMA) 20 -co-NAS).

[0370] Step 3: Preparation of hydrophilic segments via active free radical reaction

[0371] Atom transfer radical polymerization (ATRP) was also employed. The initiator was 6-Arm-P-(LMA). 20 -co-NAS). The hydrophilic monomer is OEGMA, and the average number of repeating polyethylene glycol units is approximately 4.5. The catalyst is copper bromide (CuBr2). The copper ion ligand is tris(2-dimethylaminoethyl)amine (me6tren). The solvent for NMR spectroscopy monitoring and calibration is anisole.

[0372] First, 6-Arm-P-(LMA) 20 0.61 g (0.1 mmol) of 6-Arm-P-(LMA), 1.62 g (5.4 mmol) of OEGMA, and 5.4 mg (0.024 mmol) of CuBr2 were dissolved in 5 mL of DMF, and 3 to 4 drops of anisole were added. After deoxygenation, 28 mg (0.12 mmol) of me6tren was injected. The mixture was reacted in a Shrek tube, and the conversion rate was monitored by 1H NMR spectroscopy. The reaction conditions were room temperature. During the reaction, an appropriate amount of the mixture was taken and the conversion rate was monitored by 1H NMR spectroscopy. When the conversion rate reached 94.6%, oxygen was introduced to stop the reaction. After the reaction, the DMF was evaporated, and then redissolved in tetrahydrofuran and CuBr2 was removed by passing it through a neutral alumina column. The mixture was precipitated in n-hexane and dried under vacuum at room temperature for 24 hours to obtain 1.93 g of intermediate polymer. The molecular formula of the intermediate polymer is 6-Arm-P-(LMA). 20 -co-NAS)-b-OEGMA 51 .

[0373] Step 4: Modify the intermediate polymer with fluorescent agent via substitution reaction

[0374] The intermediate polymer 6-Arm-P-(LMA) participated in the substitution reaction. 20 -co-NAS)-b-OEGMA 51 And the second unit, wherein the second unit is IR-808-NH2, wherein the fluorescent agent is a derivative of IR-783, and the second reactive group is an amino group.

[0375] 0.95g of 6-Arm-P-(LMA) 20 -co-NAS)-b-OEGMA 51 0.05 mmol of IR-808-NH2 and 5 mg of IR-808-NH2 (0.005 mmol) were dissolved in 10 mL of DMF, and 1 mL of triethylamine was added to create an alkaline environment. The mixture was stirred at room temperature in the dark for 24 hours. After the reaction was complete, 2 mL of deionized water was added to the mixture, and it was placed in a 5 kDa dialysis bag and dialyzed against deionized water for 48 hours to remove unreacted IR-808-NH2. After complete drying, 0.9 g of 6-Arm-P-(LMA) was obtained. 20 -co-(A-IR-808))-b-OEGMA 51 In this process, the substitution reaction between the first and second reactive groups on the intermediate polymer is a probabilistic substitution reaction with a reaction probability of approximately 28.6%. This allows the fluorescent agent to be probabilistically modified into the intended functional unit through the substitution reaction, while the unmodified fluorescent agent in the intended functional unit becomes the unit to be modified.

[0376] In the functional formulation of Example Sixteen, the average particle size of the polymer hydrate groups was 86.3 nanometers. The rest of the functional formulation in Example Sixteen was the same as that in Example Two.

[0377] The preparation method of the functional formulation in Example 16 is the same as that in Example 2.

[0378] See Figure 29a and Figure 29b , Figure 29a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example Sixteen after intravenous injection in mice. Figure 29b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example Sixteen into mice. The injection dosage and method of the functional formulation of Example Sixteen are the same as those in Example Two. Figure 29aAs shown, the functional formulation of Example Sixteen produced a fluorescent signal in the gallbladder (6) 5 minutes after injection, and clear fluorescent imaging of the bile duct was achieved 10 minutes after injection; a fluorescent signal appeared in the bladder (16) 5 minutes after injection; a fluorescent signal appeared in the duodenum (8) 15 minutes after injection; a fluorescent signal appeared in the small intestine (9) 30 minutes after injection; and a fluorescent signal appeared in the colon (10) 120 minutes after injection. Figure 29b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0379] Example 17

[0380] In Example 17, the polymer single chain includes a hydrophilic segment and a hydrophobic segment of block copolymerization. The hydrophilic segment includes multiple hydrophilic links, the polymeric group of which is methacrylate, and the average number of repeating units of polyethylene glycol is approximately 4.5. The hydrophobic segment includes multiple hydrophobic links, the polymeric group of which is methacrylate, and the hydrophobic group is hexyl. For the polymer, at least a portion of the polymer single chain includes functional links embedded in the hydrophobic segment, each functional link being conjugated with a fluorescent agent, the molar ratio of the fluorescent agent to the polymer single chain being 1:30, the polymeric group of the functional link being acrylate, and the fluorescent agent being indocyanine green. At least a portion of the polymer single chain includes a segment to be modified embedded in the hydrophobic segment. The segment to be modified is conjugated with a first reactive group, which is used to conjugate the fluorescent agent through a substitution reaction or coupling reaction, the molar ratio of the first reactive group to the polymer single chain being 1:1.3, and the polymeric group of the segment to be modified being acrylate. In this embodiment, the first reactive group is an amine group.

[0381] In Example 17, the polymer molecular formula is P-OEGMA 50 -b-(HMA 21 -co-(MA-ICG)). In the polymer, the average number of hydrophilic segments is 50, and the number of repeating polyethylene glycol units in the hydrophilic segments is approximately 4.5. The average number of hydrophobic segments is 21, and the hydrophobic group in the hydrophobic segments is hexyl. On average, there is one functional segment in every 30 polymer single chains. The fluorescent agent conjugated to the functional segments is indocyanine green or a derivative thereof. In Example 17, the first molecular weight of the polymer is 3600, and the second molecular weight of the polymer is 18600.

[0382] The preparation method of the polymer in Example 17 includes step 1 of preparing hydrophilic segments, step 2 of preparing hydrophobic segments with the desired modified segments, and step 3 of modifying the intermediate polymer with a fluorescent agent. Step 1 is similar to step 1 in Example 1. Step 3 is similar to step 3 in Example 13. Step 2, which differs from other examples, is as follows:

[0383] Step 2: Prepare hydrophobic segments of the chain to be modified by polymerizing the polymer via living radical polymerization:

[0384] The chain transfer agent is P-OEGMA 50 The hydrophobic monomer is HMA. The monomer to be modified is 2-aminoethyl methacrylate hydrochloride (AMA), where the polymeric group is methacrylate and the first reactive group is an amine group. The initiator is azobisisobutyronitrile (AIBN). The calibration solvent for 1H NMR spectroscopy monitoring is anisole.

[0385] First, P-OEGMA 50 1.5 g (0.1 mmol), 426 mg (2.5 mmol) HMA, 17 mg (0.1 mmol) AMA, and 3.3 mg AIBN were dissolved in 15 mL of LDM, and 3 to 4 drops of anisole were added. The mixture was reacted in a Shrek tube, and the conversion was monitored by 1H NMR spectroscopy. The reaction conditions were: deoxygenation followed by heating to 70 °C. During the reaction, an appropriate amount of the mixture was taken and the conversion was monitored by 1H NMR spectroscopy. The reaction was stopped when oxygen was introduced when the conversion reached 84.3%. After the reaction, the mixture was cooled to room temperature and evaporated to dryness. It was then reconstituted with 15 mL of tetrahydrofuran, and the supernatant was removed by precipitation in n-hexane. After vacuum drying at room temperature, 1.79 g of intermediate polymer was obtained. The intermediate polymer was P-OEGMA. 50 -b-(HMA 21 -co-AMA). The hydrophobic monomer forms a hydrophobic link within the hydrophobic segment, and the monomer to be modified forms a pre-modified link within the hydrophobic segment.

[0386] In the functional formulation of Example 17, the average particle size of the polymer hydrate groups was 42.6 nanometers. The rest of the functional formulation in Example 17 was the same as that in Example 2.

[0387] The preparation method of the functional formulation in Example 17 is the same as that in Example 2.

[0388] See Figure 30a and Figure 30b , Figure 30a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 17 after intravenous injection in mice. Figure 30b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 17 into mice. The injection dosage and method of the functional formulation of Example 17 were the same as those in Example 2. Figure 30aAs shown, the functional formulation of Example 17 showed a fluorescent signal in the gallbladder (6) 10 minutes after injection, and clear fluorescence imaging of the bile duct was achieved 30 minutes after injection; a fluorescent signal appeared in the bladder (16) 5 minutes after injection; a fluorescent signal appeared in the duodenum (8) 15 minutes after injection; a fluorescent signal appeared in the small intestine (9) 30 minutes after injection; and a fluorescent signal appeared in the colon (10) 120 minutes after injection. Figure 30b As shown, 5 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver (7) showed almost no fluorescence signal, and the colon (10) still showed fluorescence signals.

[0389] Example 18

[0390] The polymer in Example 18 has the molecular formula P-PEGMA. 53 -b-(EMA 68 -co-(MA-IR-808)). PEGMA characterizes a hydrophilic monomer with an average number of 6 to 8 repeating units in polyethylene glycol (the product is commercially available, and the nominal average number of repeating units in polyethylene glycol is 6-8, specifically around 6.2 as measured). The hydrophobic group of the hydrophobic chain segment is ethyl. The average number of hydrophilic chains in the polymer single chain is 53, and the average number of hydrophobic chains in the polymer single chain is 68. The molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the average number of repeating units in polyethylene glycol is approximately 6.2; the first molecular weight of the polymer is 7800; and the second molecular weight of the polymer is 26900. The remaining parts of the polymer in Example 18 are the same as in Example 2.

[0391] In the polymer preparation method of Example 18, the hydrophobic monomer used is ethyl methacrylate (EMA), whose hydrophobic group is ethyl. The hydrophilic monomer is PEGMA. The rest of the polymer preparation method is similar to that of Example 2.

[0392] In the functional formulation of Example 18, the average particle size of the polymer hydrate groups is 173.2 nanometers. The rest of the functional formulation of Example 18 is the same as that of Example 2.

[0393] The preparation method of the functional formulation in Example 18 is the same as that in Example 2.

[0394] See Figure 31a and Figure 31b , Figure 31a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 18 after intravenous injection in mice; Figure 31b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 18 into mice. The injection dosage and method of the functional formulation of Example 18 are the same as those in Example 2. Figure 31aAs shown, the functional formulation of Example 18 showed a fluorescent signal in the gallbladder 6 15 minutes after injection, and clear fluorescence imaging of the bile duct 20 minutes after injection; fluorescent signals appeared in the duodenum 8 and small intestine 9 30 minutes after injection. Figure 31b As shown, 2 hours after injection, the bile duct was still clearly fluorescently imaged with strong fluorescence signals, the liver 7 showed almost no fluorescence signal, and the small intestine 9 still showed fluorescence signals.

[0395] Example 19

[0396] The polymer in Example 19 has the molecular formula P-(MA-PEG8). 47 -b-(LMA 20 -co-(MA-IR-808)). MA-PEG8 represents a hydrophilic monomer with an average of 8 repeating units in polyethylene glycol, specifically octaethylene glycol monomethyl ether methacrylate. The hydrophobic group of the hydrophobic linkage is dodecyl. The average number of hydrophilic linkages in the polymer single chain is 47, and the average number of hydrophobic linkages in the polymer single chain is 20. The molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the average number of repeating units in polyethylene glycol is 8; the first molecular weight of the polymer is 5100; and the second molecular weight of the polymer is 26300.

[0397] The rest of the polymer in Example 19 is the same as in Example 2.

[0398] The preparation method of the polymer is similar to that in Example 2.

[0399] In the functional formulation of Example 19, the average particle size of the polymer hydrate groups is 93.5 nanometers. The rest of the functional formulation of Example 19 is the same as that of Example 2.

[0400] The preparation method of the functional formulation in Example 19 is the same as that in Example 2.

[0401] See Figure 32a and Figure 32b , Figure 32a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 19 after intravenous injection in mice; Figure 32b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 19 into mice. The injection dosage and method of the functional formulation of Example 19 were the same as those in Example 2. Figure 32a As shown, the functional formulation of Example 19 showed fluorescent signals in the gallbladder 6 and duodenum 8 30 minutes after injection, with a significant decrease in liver signal. The bile duct was clearly fluorescently imaged 45 minutes after injection. In this mouse, the duodenum was embedded in the posterior part of the liver; 2 hours after injection, due to mouse activity, a significant fluorescent signal in the small intestine 9 could be observed. Figure 32bAs shown, when the abdominal cavity was opened 5 hours after injection, the bile ducts were still clearly fluorescently imaged with strong fluorescence signals, while the liver 7 showed almost no fluorescence signal.

[0402] Example 20

[0403] The polymer in Example 20 has the molecular formula P-(MA-PEG4). 74 -b-(LMA 20 -co-(MA-IR-808)). MA-PEG4 represents a hydrophilic monomer with an average of 4 repeating units in polyethylene glycol, specifically tetraethylene glycol monomethyl ether methacrylate. The hydrophobic group of the hydrophobic linkage is dodecyl. The average number of hydrophilic linkages in the polymer single chain is 74, and the average number of hydrophobic linkages in the polymer single chain is 20. The molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the average number of repeating units in polyethylene glycol is 4; the first molecular weight of the polymer is 5100; and the second molecular weight of the polymer is 20500.

[0404] The rest of the polymer in Example 20 is the same as in Example 2.

[0405] The preparation method of the polymer is similar to that in Example 2.

[0406] In the functional formulation of Example 20, the average particle size of the polymer hydrate groups is 89.3 nanometers. The rest of the functional formulation of Example 20 is the same as that of Example 2.

[0407] The preparation method of the functional formulation in Example 20 is the same as that in Example 2.

[0408] See Figure 33a and Figure 33b , Figure 33a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 20 after intravenous injection in mice; Figure 33b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 20 into mice. The injection dosage and method of the functional formulation of Example 20 are the same as those in Example 2. Figure 33a As shown, in Example 20, the functional formulation produced a fluorescent signal in the gallbladder (6) 15 minutes after injection, and a significant decrease in liver signal 30 minutes after injection, with clear fluorescence imaging of the bile ducts; in this mouse, the duodenum was embedded in the posterior part of the liver, and a fluorescent signal appeared in the small intestine (9) 2 hours and 45 minutes after injection. Figure 33b As shown, when the abdominal cavity was opened 5 hours after injection, the gallbladder 6 and small intestine 9 were still clearly fluorescently imaged with strong fluorescence signals, while the liver 7 showed almost no fluorescence signal.

[0409] Example 21

[0410] The polymer in Example 20 has the molecular formula P-(MA-PEG4). 69 -b-(LMA 30 -co-(MA-IR-808)). MA-PEG4 represents a hydrophilic monomer with an average of 4 repeating units in polyethylene glycol, specifically tetraethylene glycol monomethyl ether methacrylate. The hydrophobic group of the hydrophobic linkage is dodecyl. The average number of hydrophilic linkages in the polymer single chain is 69, and the average number of hydrophobic linkages in the polymer single chain is 30. The molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the average number of repeating units in polyethylene glycol is 4; the first molecular weight of the polymer is 7700; and the second molecular weight of the polymer is 26800.

[0411] The rest of the polymer in Example 21 is the same as in Example 2.

[0412] The preparation method of the polymer is similar to that in Example 2.

[0413] In the functional formulation of Example 21, the average particle size of the polymer hydrate groups is 132.7 nanometers. The rest of the functional formulation of Example 2 is the same as that of Example 2.

[0414] The preparation method of the functional formulation in Example 21 differs from that in Example 2. Specifically, in this example, 2g of polymer was added to 15mL of phosphate buffer and ultrasonically vibrated for 30 minutes. The polymer swelled slightly, and some of it was dispersed in the phosphate buffer. After standing in the dark at room temperature for 12 days, the polymer was completely dispersed in the phosphate buffer, thus obtaining the functional formulation. If the standing time is less than 12 days, the completely dispersed portion can be extracted using the method in Comparative Example 10 or Comparative Example 11 to obtain the functional formulation.

[0415] See Figure 34a and Figure 34b , Figure 34a The image shown is a transdermal whole-body fluorescence imaging image of the functional formulation of Example 21 after intravenous injection in mice; Figure 34b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the functional formulation of Example 21 into mice. The injection dosage and method of the functional formulation of Example 21 are the same as those in Example 2. Figure 34a As shown, in Example 21, the functional formulation showed a significant decrease in liver signal (7) 30 minutes after injection, a strong signal in gallbladder (6) 45 minutes after injection, clear fluorescence imaging of the bile duct, a noticeable signal in duodenum (8), and a noticeable signal in small intestine (9) 120 minutes after injection. Figure 34b As shown, when the abdominal cavity was opened 5 hours after injection, the gallbladder 6 was still clearly fluorescently imaged with obvious fluorescence signal, while the liver 7 showed almost no fluorescence signal.

[0416] Comparative Example 1

[0417] The polymer in Comparative Example 1 has the molecular formula P-(MA-PEG3). 80 -b-LMA 32 It comprises hydrophilic and hydrophobic segments. The hydrophilic segments include multiple hydrophilic chain segments, each containing three repeating units of polyethylene glycol. The hydrophilic monomer (MA-PEG) is triethylene glycol monomethyl ether methacrylate, and the hydrophobic segments include multiple hydrophobic chain segments, with the hydrophobic group being dodecyl. The polymer in Comparative Example 1 has a first molecular weight of 8200 and a second molecular weight of 28200.

[0418] See Figure 35 , Figure 35 An image showing the polymer of Comparative Example 1 dispersed in water is displayed. Figure 35 As shown, the polymer in Comparative Example 1 cannot be dispersed in water. Therefore, the polymer in Comparative Example 1 cannot be used to prepare functional formulations.

[0419] Comparative Example 2

[0420] The polymer in Comparative Example 2 has the molecular formula P-(MA-PEG4). 80 -b-LMA 32 It comprises hydrophilic and hydrophobic segments. The hydrophilic segments include multiple hydrophilic units, with each polyethylene glycol repeating unit having four units. The hydrophilic monomer (MA-PEG4) is tetraethylene glycol monomethyl ether methacrylate, and the hydrophobic segments include multiple hydrophobic units, with the hydrophobic group being dodecyl. The polymer in Comparative Example 2 has a first molecular weight of 8200 and a second molecular weight sum of 31700.

[0421] See Figure 36 , Figure 36 The image shown illustrates the state of the polymer from Comparative Example 2 dispersed in water after ultrasonic agitation for 30 minutes. Figure 36 As shown, at this time, the polymer of Comparative Example 2 can only be partially dispersed in water for a short period of time.

[0422] Comparative Example 3

[0423] The polymer in Comparative Example 3 has the molecular formula P-(MA-PEG9). 46 -b-(LMA 10The polymer (-co-(MA-IR-808)) comprises hydrophilic and hydrophobic segments. The hydrophilic segments contain multiple hydrophilic units, with an average of approximately 9.1 repeating polyethylene glycol units. The hydrophilic monomer is commercially available polyethylene glycol monomethyl ether methacrylate with an average molecular weight of 500. The hydrophobic segments contain multiple hydrophobic units, with dodecyl groups in the hydrophobic groups. Functional segments are embedded within the hydrophobic segments, and the fluorescent agent is IR-808, a derivative of IR-783. The polymer in Comparative Example 3 has a first molecular weight of 2600 and a second molecular weight of 25600. The hydrated polymer particles in the comparative formulation have a particle size of 45.7 nm.

[0424] See Figure 37a and Figure 37b . Figure 37a Transdermal whole-body fluorescence imaging images of the comparative formulation in mice after intravenous injection are shown in Comparative Example 3. Figure 37b The image shows fluorescence imaging of the liver and gallbladder after intravenous injection of the comparative formulation in mice, as shown in Comparative Example 3. Figure 37a and Figure 37b As shown, 3 hours after injection, the liver (7) of the mice still showed a strong fluorescence signal, while the gallbladder (6) showed an indistinct fluorescence signal, making it impossible to obtain clear fluorescence imaging of the bile duct.

[0425] Comparative Example 4

[0426] The polymer in Comparative Example 4 has the molecular formula P-(MA-PEG). 19 ) 24 -b-(LMA 10 The polymer (-co-(MA-IR-808)) comprises hydrophilic and hydrophobic segments. The hydrophilic segments contain multiple hydrophilic units, with an average of approximately 19.3 repeating polyethylene glycol units. The hydrophilic monomer is commercially available polyethylene glycol monomethyl ether methacrylate with an average molecular weight of 950. The hydrophobic segments contain multiple hydrophobic units, with dodecyl groups within the hydrophobic groups. Functional segments are embedded within the hydrophobic segments, and the fluorescent agent is IR-808, a derivative of IR-783. The polymer in Comparative Example 4 has a first molecular weight of 2600 and a second molecular weight of 25400. The hydrated polymer particles in the comparative formulation have a particle size of 43.5 nm.

[0427] See Figure 38a and Figure 38b . Figure 38a Transdermal whole-body fluorescence imaging images of the comparative formulation in mice after intravenous injection are shown in Comparative Example 4. Figure 38b The image shows fluorescence imaging of the liver and gallbladder after intravenous injection of the comparative formulation in mice, as shown in Comparative Example 4. Figure 38a and Figure 38bAs shown, 3 hours after injection, the liver 7 of the mouse still showed a strong fluorescence signal, while the fluorescence signal of the gallbladder 6 was not obvious, and clear fluorescence imaging of the gallbladder 6 could not be performed.

[0428] Comparative Example 5

[0429] The polymer in Comparative Example 5 has the molecular formula P-OEGMA. 61 -b-(LMA5-co-(MA-IR-808)), which includes hydrophilic and hydrophobic segments. The hydrophilic segments consist of multiple hydrophilic units, with an average of approximately 4.5 repeating polyethylene glycol units in each segment. The hydrophobic segments consist of multiple hydrophobic units, with dodecyl groups in each segment. Functional segments are embedded in the hydrophobic segments, and the fluorescent agent is IR-808, a derivative of IR-783. The polymer in Comparative Example 5 has a first molecular weight of 1300 and a second molecular weight of 19600. The hydrated polymer particles in the comparative formulation have a particle size of 28.9 nm.

[0430] See Figure 39a and Figure 39b . Figure 39a The image shows a transdermal whole-body fluorescence imaging of the comparative formulation in mice after intravenous injection in a supine position. Figure 39b The image shows a transdermal whole-body fluorescence imaging image of the comparative formulation of Comparative Example 5 in mice after intravenous injection in a prone position. Figure 39a and Figure 39b As shown, before 6 hours after injection, the bladder 16 and kidney 17 of the mice consistently showed strong fluorescence signals, while the fluorescence signal of gallbladder 6 was not obvious, making it impossible to obtain clear fluorescence imaging of the bile duct.

[0431] Comparative Example 6

[0432] The polymer in Comparative Example 6 has the molecular formula P-OEGMA. 42 -b-(LMA 11 The polymer (-co-(MA-IR-808)) comprises hydrophilic and hydrophobic segments. The hydrophilic segments include multiple hydrophilic units, with an average of approximately 4.5 repeating polyethylene glycol units per segment. The hydrophobic segments also include multiple hydrophobic units, with dodecyl groups as the hydrophobic groups. Functional segments are embedded within the hydrophobic segments, and the fluorescent agent is IR-808, a derivative of IR-783. The polymer in Comparative Example 6 has a first molecular weight of 2800 and a second molecular weight of 15400. The hydrated polymer particles in the comparative formulation have a particle size of 37.2 nm.

[0433] See Figure 40 . Figure 40 Transdermal whole-body fluorescence imaging images of the comparative formulation of Comparative Example 6 after intravenous injection in mice are shown. Figure 40As shown, bladder 16 still had a strong fluorescence signal 5 hours after injection, liver 7 maintained a fluorescence signal from injection to 1 hour, and gallbladder 6 had an inconspicuous fluorescence signal, making it impossible to obtain clear fluorescence imaging of gallbladder 6.

[0434] Comparative Example 7

[0435] The polymer in Comparative Example 7 has the molecular formula P-OEGMA. 50 -b-(LMA 35 The polymer (-co-(MA-IR-808)) comprises hydrophilic and hydrophobic segments. The hydrophilic segments include multiple hydrophilic units, with an average of approximately 4.5 repeating polyethylene glycol units within each segment. The hydrophobic segments also include multiple hydrophobic units, with dodecyl groups as the hydrophobic groups. Functional segments are embedded within the hydrophobic segments, and the fluorescent agent is IR-808, a derivative of IR-783. The polymer in Comparative Example 7 has a first molecular weight of 8900 and a second molecular weight of 23900. The hydrated polymer particles in the comparative formulation have a particle size of 452.6 nm.

[0436] See Figure 41a and Figure 41b . Figure 41a Transdermal whole-body fluorescence imaging images of the comparative formulation of Comparative Example 7 after intravenous injection in mice are shown. Figure 41b The image shows fluorescence imaging of the liver and gallbladder after intravenous injection of the comparative formulation of Comparative Example 7 in mice. Figure 41a and Figure 41b As shown, the liver (7) of mice still showed a strong fluorescence signal 3 hours after injection, while the gallbladder (6) only showed a significant fluorescence signal 120 minutes after injection. Therefore, it was impossible to quickly obtain clear fluorescence imaging of the bile duct.

[0437] Comparative Example 8

[0438] The polymer in Comparative Example 8 has the molecular formula P-OEGMA. 76 -b-(LMA 23 The polymer (-co-(MA-IR-808)) comprises hydrophilic and hydrophobic segments. The hydrophilic segments include multiple hydrophilic units, with an average of approximately 4.5 repeating polyethylene glycol units within each segment. The hydrophobic segments also include multiple hydrophobic units, with dodecyl groups as the hydrophobic groups. Functional segments are embedded within the hydrophobic segments, and the fluorescent agent is IR-808, a derivative of IR-783. The polymer in Comparative Example 8 has a first molecular weight of 5900 and a second molecular weight of 28700. The hydrated polymer particles in the comparative formulation have a particle size of 274.9 nm.

[0439] See Figure 42a and Figure 42b . Figure 42aTransdermal whole-body fluorescence imaging images of the comparative formulation of Comparative Example 8 after intravenous injection in mice are shown. Figure 42b The image shows fluorescence imaging of the liver and gallbladder after intravenous injection of the comparative formulation of Comparative Example 8 in mice. Figure 42a and Figure 42b As shown, the gallbladder 6 of the mouse showed a fluorescent signal 10 minutes after injection, and the liver 7 still showed a strong fluorescent signal up to 5 hours after injection, so it was impossible to quickly obtain clear fluorescent imaging of the bile duct.

[0440] Comparative Example 9

[0441] The polymer in Comparative Example 9 has the molecular formula P-OEGMA. 76 -co-(MA-IR-808) consists only of hydrophilic segments, each comprising multiple hydrophilic chain segments. The average number of repeating polyethylene glycol units within each hydrophilic chain segment is approximately 4.5. Functional chain segments are embedded within the hydrophilic segments, and the fluorescent agent is IR-808, a derivative of IR-783. The polymer in Comparative Example 9 has a first molecular weight of 0 and a second molecular weight of 25200. The hydrated polymer particles in the comparative formulation have a particle size of 96.3 nanometers.

[0442] See Figure 43 . Figure 43 The image shows transdermal whole-body fluorescence imaging of the comparative formulation of Comparative Example 9 after intravenous injection in mice. Figure 43 As shown, five minutes after injection, both liver 7 and bladder 16 in mice showed fluorescent signals. The fluorescent signal in liver 7 basically disappeared after 30 minutes, while the fluorescent signal in bladder 16 lasted for at least one hour. There was no fluorescent signal in gallbladder 6.

[0443] Comparative Example 10

[0444] The polymer in Comparative Example 10 has the molecular formula P-((MA-PEG4)). 61 -co-(MA-PEG3) 26 )-b-(LMA 16-co-(MA-IR-808)). MA-PEG4 represents a hydrophilic monomer with an average of 4 repeating units in polyethylene glycol, specifically tetraethylene glycol monomethyl ether methacrylate. MA-PEG3 represents a hydrophilic monomer with an average of 3 repeating units in polyethylene glycol, specifically triethylene glycol monomethyl ether methacrylate. The hydrophobic group of the hydrophobic segment is dodecyl. The average number of hydrophilic segments in the polymer single chain is 87, and the average number of hydrophobic segments in the polymer single chain is 16. The molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the average number of repeating units in polyethylene glycol is approximately 3.7; the first molecular weight of the polymer is 4100; and the second molecular weight of the polymer is 27000. The remaining parts of the polymer in Comparative Example 10 are the same as in Example 2. When preparing the hydrophilic segments, the hydrophilic monomers are tetraethylene glycol monomethyl ether methacrylate (MA-PEG4) and triethylene glycol monomethyl ether methacrylate (MA-PEG3). The chain transfer agent was 4-cyano-4-(thiobenzoyl)valerate (CTA). The initiator was azobisisobutyronitrile (AIBN). Anisole was used as the solvent for calibration by 1H NMR spectroscopy. First, 56 mg (0.2 mmol) of CTA, 3.65 g (13.2 mmol) of MA-PEG4, 1.3 g (5.6 mmol) of MA-PEG3, and 3.3 mg (0.02 mmol) of AIBN were dissolved in 15 mL of dioxane, and 3 to 4 drops of anisole were added. The mixed solution was reacted in a Shrek tube, and the conversion rate was monitored by 1H NMR spectroscopy. The reaction conditions were: after deoxygenation, the temperature was raised to 70 °C. During the reaction, an appropriate amount of the mixed solution was taken and the conversion rate was monitored by 1H NMR spectroscopy. When the conversion rate reached 92.3%, oxygen was introduced to stop the reaction. After the reaction, the solution was cooled to room temperature, and the supernatant was removed by precipitation in n-hexane. After vacuum drying at room temperature, 4.37 g of hydrophilic segments were obtained. The molecular formula of the hydrophilic segment is P-((MA-PEG4)61-co-(MA-PEG3)26). The remaining steps of polymer preparation are similar to those in Example 2. When the polymer is dispersed in water, it can only be partially dispersed (the dispersed mass accounts for less than 50% of the total polymer mass). After centrifugation (10000 rpm, 5 min), the polymer dispersion is collected, and the supernatant is then passed through a 220 nm needle filter to remove a small amount of undispersed polymer that was not centrifuged out, resulting in a comparative formulation. In the comparative formulation, the polymer hydrated aggregate particle size is 178.5 nm, and the average number of repeating units of polyethylene glycol in the comparative formulation is still about 3.7.

[0445] See Figure 44a and Figure 44b , Figure 44a Transdermal whole-body fluorescence imaging images of the comparative formulation of Comparative Example 10 after intravenous injection in mice are shown. Figure 44bThe image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the comparative formulation of Comparative Example 10 into mice. The injection dose and method of the comparative formulation of Comparative Example 10 were the same as in Example 2. Figure 44a As shown, in Comparative Example 10, the control formulation showed a fluorescent signal in the gallbladder (6) 10 minutes after injection, but the signal in the liver (7) did not show significant attenuation. Figure 44b As shown, when the abdominal cavity was opened 5 hours after injection, fluorescent signals were observed in both gallbladder 6 and liver 7. However, the fluorescent signal in gallbladder 6 was not significantly greater than that in liver 7, indicating that the polymer remained in liver 7 and was not rapidly metabolized. This situation is not conducive to fluorescent surgical navigation of the biliary tract.

[0446] Comparative Example 11

[0447] The polymer in Comparative Example 11 has the molecular formula P-((MA-PEG4)). 28 -co-(MA-PEG3) 65 )-b-(LMA 16 -co-(MA-IR-808)). MA-PEG4 represents a hydrophilic monomer with an average of 4 repeating units in polyethylene glycol, specifically tetraethylene glycol monomethyl ether methacrylate. MA-PEG3 represents a hydrophilic monomer with an average of 3 repeating units in polyethylene glycol, specifically triethylene glycol monomethyl ether methacrylate. The hydrophobic group of the hydrophobic linkage is dodecyl. The average number of hydrophilic linkages in the polymer single chain is 93, and the average number of hydrophobic linkages in the polymer single chain is 16. The molar ratio of fluorescent agent to polymer single chain is 1:30. In the polymer, the average number of repeating units in polyethylene glycol is approximately 3.3; the first molecular weight of the polymer is 4100; and the second molecular weight of the polymer is 27000. The remaining parts of the polymer in Comparative Example XI are the same as those in Comparative Example X. When the polymer is dispersed in water, it can only be partially dispersed (the mass of the dispersed part accounts for less than 50% of the total mass of the polymer). After centrifugation (10000 rpm, 5 min), the polymer dispersion is then collected, and a small amount of undispersed polymer that was not centrifuged is removed by passing it through a 220 nm needle filter to obtain the control formulation. In the control formulation, the polymer hydrated aggregate particle size is 164.3 nm.

[0448] See Figure 45a and Figure 45b , Figure 45a The image shown is a transdermal whole-body fluorescence imaging image of the comparative formulation of Comparative Example 11 after intravenous injection in mice. Figure 45b The image shows a fluorescence imaging image of the liver and gallbladder after intravenous injection of the comparative formulation of Comparative Example 11 into mice. The injection dose and method of the comparative formulation of Comparative Example 11 were the same as in Example 2. Figure 45aAs shown, in Comparative Example 11, the control formulation showed a fluorescent signal in gallbladder 6 15 minutes after injection, but the signal in liver 7 did not show significant attenuation. Figure 45b As shown, when the abdominal cavity was opened 5 hours after injection, fluorescent signals were observed in both gallbladder 6 and liver 7. However, the fluorescent signal in gallbladder 6 was not significantly greater than that in liver 7, indicating that the polymer remained in liver 7 and was not rapidly metabolized. This situation is not conducive to fluorescent surgical navigation of the biliary tract.

[0449] In the above embodiments, the applicant achieves "block copolymerization" through block copolymerization. "Block copolymerization" refers to the linkage of two or more polymer segments with different properties. Specifically, block copolymerization in this application does not include random copolymerization. To determine whether a polymer is a block copolymer, the preferred method is detection using 1H NMR spectroscopy. Block copolymerization is indicated when the characteristic peaks of the methyl group in methacrylate or the methylene group in acrylate exhibit a bimodal distribution; it is not block copolymerization when the characteristic peaks of the methyl group in methacrylate or the methylene group in acrylate exhibit a single peak or a broad distribution. If the obtained sample is a functional formulation, the polymer is obtained by dialyzing in deionized water and then completely drying.

[0450] In the above embodiments, the applicant controls the average number of repeating units of polyethylene glycol in the polymer by controlling the number or average number of repeating units of polyethylene glycol in the hydrophilic monomer. To determine the average number of repeating units of polyethylene glycol in the polymer, the preferred method is as follows: First, hydrolyze the polymer to obtain a mixture of polyethylene glycol, an alcohol with hydrophobic groups, the main chain of the polymer single chain, and water; second, place the mixture in dichloromethane and extract the organic components to obtain a mixture of polyethylene glycol, an alcohol with hydrophobic groups, the main chain of the polymer single chain, and dichloromethane; third, remove part of the dichloromethane and place the remaining mixture in n-hexane and remove the main chain of the polymer single chain by sedimentation, with both polyethylene glycol and the alcohol with hydrophobic groups remaining in the supernatant; finally, after the supernatant is completely dried, the obtained substances are subjected to high-performance liquid chromatography to obtain substances with similar molecular weights and molecular weights. The average molecular weight of similar substances and the mass percentage of substances with similar molecular weights in the total substance are determined. The chemical structure of each substance in the substance with similar molecular weight and its molar ratio to other substances in the substance with similar molecular weight are obtained by proton nuclear magnetic resonance spectroscopy. All substances with chemical structures belonging to polyethylene glycol and the number of repeating units in these substances are identified. Based on the number of repeating units in the substance with the chemical structure of polyethylene glycol, the molar ratio of this substance to other substances with similar molecular weights, the average molecular weight of the substance to which it belongs, and the mass ratio of the substance to which it belongs to other substances with similar molecular weights, the average number of repeating units of polyethylene glycol in the polymer is determined.

[0451] The above embodiments control the first molecular weight by controlling the degree of polymerization of the hydrophobic monomer, and control the second molecular weight by controlling the degree of polymerization of the hydrophobic monomer and the hydrophilic monomer, respectively. To determine the first and second molecular weights in the polymer, the preferred method is as follows: First, obtain the average molecular weight of the polymer using gel permeation chromatography; second, obtain the chemical structures of all hydrophobic groups, the chemical structures of the polymeric groups, the molar ratios of all hydrophobic groups to all polyethylene glycols, and the molar ratios of all polyethylene glycols to the polymeric groups using 1H NMR spectroscopy. Based on the average molecular weight of the polymer, the average number of repeating units in the polyethylene glycol, the chemical structures of the hydrophobic groups and polymeric groups, the molar ratios of all hydrophobic groups to all polyethylene glycols, and the molar ratios of all polyethylene glycols to all polymeric groups, the average molecular weight of the hydrophilic ends of all polymer single chains and the first molecular weight are obtained, and thus the second molecular weight is obtained. Other detection methods can also be used if they are more accurate and easier.

[0452] It should be noted that in the above embodiments and comparative examples, the transdermal whole-body fluorescence imaging images of mice should be observed and analyzed together with the hepatobiliary anatomical fluorescence imaging images. Due to the anatomical location of the gallbladder, it may be obscured by the xiphoid process in certain body positions, making the gallbladder fluorescence imaging appear unclear. However, when actually applied to biliary fluorescence imaging surgical navigation, the fluorescence imaging is closer to the hepatobiliary anatomical fluorescence imaging images.

[0453] Furthermore, the above embodiments and comparative examples were purposefully extracted from a large amount of experimental data and do not represent that only the experimental data of the above embodiments and comparative examples exist. The description of the above embodiments and comparative examples is only to illustrate the basis for the applicant's selection of relevant parameters.

[0454] In the above examples and comparative examples, a high fluorescence signal intensity in the bladder or kidney indicates that the functional agent is metabolized more by the kidneys. A low fluorescence signal intensity in the bladder or kidney indicates that the functional agent is metabolized less by the kidneys. No fluorescence signal in the bladder or kidney indicates that the functional agent is not metabolized by the kidneys.

[0455] In the above examples and comparative examples, if the liver fluorescence signal is strong and lasts for a long time, it means that the functional agent has been phagocytosed by hepatocytes, and this is considered as unclear fluorescence imaging of the gallbladder.

[0456] In the above embodiments and comparative examples, if the fluorescence signal of the gallbladder is significantly stronger than that of the liver, it can be considered that the bile duct is clearly fluorescently imaged. In particular, hepatobiliary anatomical fluorescence imaging images are more characterizing of whether the gallbladder is clearly fluorescently imaged.

[0457] In the above examples and comparative examples, since the bile ducts of mice are thin and difficult to distinguish, the strong fluorescence signal of the mouse gallbladder is considered to be able to clearly fluoresce the bile ducts and gallbladders of large mammals.

[0458] See Table 1, which provides a comprehensive comparison of all the above embodiments and comparative examples.

[0459] Table 1: Comprehensive Comparison of Parameters and Experimental Results between Twenty-One Examples and Eleven Comparative Examples

[0460]

[0461]

[0462] Example 16 demonstrates that even a star-shaped topological structure of the polymer single chain allows for clear fluorescence imaging of the gallbladder or bile duct. Examples 14 and 15 demonstrate that a linear topological structure with three segments in the polymer single chain allows for clear fluorescence imaging of the gallbladder or bile duct. Other examples demonstrate that a linear topological structure with two segments in the polymer single chain allows for clear fluorescence imaging of the gallbladder or bile duct. Therefore, the topological structure of the polymer has little impact on whether the functional formulation is rapidly metabolized by the liver and reaches the bile duct.

[0463] As demonstrated in Examples 13 and 15, the functional segments are located in hydrophilic segments, enabling clear fluorescence imaging of the gallbladder and / or bile ducts. As demonstrated in other examples, the functional segments are located in hydrophobic segments, enabling clear fluorescence imaging of the gallbladder and / or bile ducts.

[0464] As shown in Example 16, when the living radical polymerization reaction is an atom transfer radical polymerization reaction during polymer preparation, a polymer that allows for clear fluorescence imaging of the gallbladder or bile duct can be prepared. As shown in other examples, when the living radical polymerization reaction is a reversible addition-fragmentation chain transfer polymerization reaction during polymer preparation, a polymer that allows for clear fluorescence imaging of the gallbladder or bile duct can be prepared.

[0465] As can be seen from Examples 13 and 17, when the polymer group is an acrylate group, a living free radical polymerization reaction can be achieved. As can be seen from other examples, when the polymer group is a methacrylate group, a living free radical polymerization reaction can be achieved.

[0466] As can be seen from Comparative Example 1, when the average number of repeating units in polyethylene glycol is 3, the polymer cannot be dispersed in water or aqueous solution and cannot be used to prepare functional formulations.

[0467] As can be seen from Comparative Example 2, Example 20 and Example 21, when the average number of repeating units of polyethylene glycol is 4, the polymer can only be partially dispersed in water or aqueous solution in the short term, but can be completely dispersed in the long term.

[0468] As can be seen from Comparative Examples 3 and 4, even if the first and second molecular weights of the polymer are appropriate, the functional formulation is still easily phagocytosed by hepatocytes when the average number of repeating units of polyethylene glycol is greater than 8.

[0469] Comparative Examples 10 and 11 show that even when the first and second molecular weights are appropriate, functional formulations are still easily phagocytosed by hepatocytes when the average number of repeating units of polyethylene glycol is less than 4.

[0470] As can be seen from Comparative Example 7, even if the average number of repeating units in polyethylene glycol and the second molecular weight of the polymer are appropriate, the functional formulation is still easily phagocytosed by hepatocytes when the first molecular weight of the polymer is greater than 7800.

[0471] As can be seen from Comparative Example 8, even if the average number of repeating units in polyethylene glycol and the first molecular weight of the polymer are appropriate, the functional formulation is still easily phagocytosed by hepatocytes when the second molecular weight of the polymer is greater than 27,100.

[0472] As can be seen from Comparative Example 5, even if the average number of repeating units of polyethylene glycol and the second molecular weight of the polymer are appropriate, the functional formulation is still easily metabolized by the kidneys when the first molecular weight of the polymer is less than 1800.

[0473] As can be seen from Comparative Example 6, even if the average number of repeating units and the first molecular weight of polyethylene glycol are appropriate, the functional formulation is still easily metabolized by the kidneys when the second molecular weight of the polymer is less than 16,800.

[0474] Based on a comprehensive analysis of the above embodiments and comparative examples, it can be seen that only when the average number of repeating units of polyethylene glycol is between 4 and 8, the first molecular weight of the polymer is between 1800 and 7800, and the second molecular weight of the polymer is between 16800 and 27100, can the functional formulation prepared by the polymer be easily and rapidly metabolized by the liver and enter the bile duct, and can the bile duct be clearly fluorescently imaged.

[0475] See Figure 46 and Figure 47 , Figure 46 and Figure 47 This illustrates the physiological structure of the mammalian liver in transporting substances to the bile duct via the bloodstream. (For example...) Figure 46 and Figure 47 As shown, the hepatic artery 18 and portal vein 19 converge into the central vein 21 via hepatic sinusoids 20. Between the hepatic sinusoids 20 and the perisinusoidal space 22, there are hepatic sinusoidal endothelial cells 23. Each hepatic sinusoidal endothelial cell 23 has a 200-nanometer opening through which substances can enter the perisinusoidal space 22. Between the perisinusoidal space 22 and the bile capillaries 24, there are hepatocytes 25, with an intercellular space of 3 nanometers according to literature. The bile capillaries 24 converge into the intrahepatic bile ducts 26, ultimately entering the extrahepatic bile ducts.

[0476] See Figure 48 , Figure 48 The image shows a polymer single-chain hydrate as simulated by molecular dynamics. (Example) Figure 48 As shown, the polymer single-chain hydrates are rod-shaped, with their diameter considered as particle size. The hydrophilic segments are exposed outwards, while the hydrophobic segments are curled inwards. The applicant believes that it is precisely within the perisinusoidal space 24 that the polymer hydrates dissociate into polymer single-chain hydrates due to the unique structure of the hepatic sinusoids and the intrahepatic blood pressure. When the particle size of the polymer single-chain hydrates is smaller than the intercellular space of hepatocytes (3 nanometers), the polymer single-chain hydrates can rapidly pass through the intercellular space and ultimately enter the bile duct.

[0477] The hydrated polymer single chains required molecular dynamics simulations. BIOVIA Materials Studio 2022 software was used for these simulations. The assembly size and morphology of individual polymer chains in water were simulated using polymer single chains. The MS Visualizer module was used for polymer molecular model construction, visualization, and output; Amorphous Cell was used to construct the polymer-water mixture model; and the Forcite module was used for model geometry optimization, molecular dynamics calculations, result analysis, and output. The constructed polymer and water molecules were then filled into an empty container, with a water filling density of 1.0 g / cm³. 3 A model of a mixture of polymer molecules and water was obtained. The box was a cuboid with dimensions of [missing information]. The energy calculation parameters are shown in Table 2, and the geometric optimization parameters are shown in Table 3.

[0478] Table 2: Energy Calculation Parameters for Polymer Single-Chain Hydrates in Molecular Dynamics Simulations

[0479]

[0480] Table 3; Geometric optimization parameters for polymer single-chain hydrates in molecular dynamics simulations

[0481]

[0482] Among them, the most important parameter in the energy calculation parameter table is the force field. The COMPASS (Condensed-phase Optimized Molecular Potentials for Atomistic Simulation Studies) series of force fields are obtained based on ab initio fitting and combined with quantum mechanical calculation results (B3LYP / 6-31G(d,p) accuracy level), which further improves the original COMPASS force field, not only covering a wider range but also having higher calculation accuracy.

[0483] In the molecular dynamics simulation, the ensemble was set to NVT, the initial velocity was adopted as Maxwell-Boltzmann distribution, the simulation temperature was 298K (25℃) and a Nosé-Hoover thermostat was used, the time step was 1fs, and the total simulation time was 5000ps.

[0484] Molecular dynamics results must be obtained after the system reaches equilibrium. Therefore, to ensure the reliability of the results, the equilibrium of the system must be determined before analysis (e.g., whether the energy and temperature change curves have reached equilibrium). Based on the stable conformation of the polymer, it is considered as a cylinder. First, the Connolly method is used to determine the volume occupied by the polymer. Then, the diameter of the equal-volume cylinder is calculated. This diameter is used as the particle size of the polymer single-chain hydrate and as the determination size for its passage through the intercellular spaces of hepatocytes.

[0485] See Table 4, which shows the polymer hydrated particle size and polymer single-chain hydrate particle size from molecular dynamics simulations for all the above examples and comparative examples.

[0486] Table 4: Polymer hydrated particle size and polymer single-chain hydrate particle size in twenty-one examples and eleven comparative examples

[0487]

[0488] As shown in Table 4, in all embodiments, the polymer single-chain hydrates that can be rapidly metabolized by the liver and reach the bile duct have a particle size of less than 3 nanometers, which is consistent with the structure of hepatic sinusoids in mammals. Since humans and other mammals have similar hepatic sinusoidal structures, the relevant results can be used to characterize the effects after application to humans.

[0489] Comparative Examples 5 and 8 show that even if the average particle size of polymer hydrates is large, the particle size of polymer single-chain hydrates can still be small. Comparative Example 4 shows that even if the average particle size of polymer hydrates is small, the particle size of polymer single-chain hydrates can still be large.

[0490] Comparative Examples 5 and 7 show that even if the second molecular weight of the polymer is large, the particle size of the polymer single-chain hydrate may still be small; even if the second molecular weight of the polymer is small, the particle size of the polymer single-chain hydrate may still be large.

[0491] As can be seen from Comparative Example 8, even if the first molecular weight of the polymer is large, the particle size of the polymer single-chain hydrate may still be small; even if the first molecular weight is small, the particle size of the polymer single-chain hydrate may still be large.

[0492] A comprehensive analysis of all examples and comparative examples reveals that the particle size of polymer single-chain hydrates is related to both the first molecular weight and the second molecular weight of the polymer. Only when the first molecular weight of the polymer is less than 7800 and the second molecular weight is less than 27100, is the particle size of the polymer single-chain hydrate less than 3 nanometers, suitable for passing through the intercellular spaces of hepatocytes.

[0493] As can be seen from Comparative Example 3, even if the particle size of the polymer single-chain hydrate is less than 3 nanometers, it will still be phagocytosed by hepatocytes because the average number of repeating units of polyethylene glycol is greater than 8.

[0494] Comparative Examples 10 and 11 show that even if the particle size of the polymer single-chain hydrate is less than 3 nanometers, it will still be phagocytosed by hepatocytes because the average number of repeating units of polyethylene glycol is less than 4.

[0495] Therefore, the parameters for functional formulations to reach the bile ducts rapidly without being phagocytosed by hepatocytes should also include the number of repeating units in polyethylene glycol. That is, only when the first molecular weight of the polymer is less than 7800, the second molecular weight of the polymer is less than 27100, and the number of repeating units in polyethylene glycol is between 4 and 8, can it be guaranteed that the functional formulation will not be phagocytosed by hepatocytes and will reach the bile ducts rapidly via liver metabolism.

[0496] Comparing Example 4 and Comparative Example 6, it can be seen that even if the first molecular weight of the polymer is relatively large, it may still be metabolized by the kidneys in large quantities. Even if the first molecular weight of the polymer is relatively small, it may still be metabolized by the kidneys in small quantities.

[0497] Of course, as can be seen from the comparison between Example 5 and Comparative Example 5, even if the second molecular weight of the polymer is large, it is still possible that a small amount will be metabolized by the kidneys; even if the second molecular weight of the polymer is small, it is still possible that a large amount will be metabolized by the kidneys.

[0498] In summary, the first and second molecular weights of the polymer together determine whether it is metabolized by the kidneys. That is, only when the first molecular weight of the polymer is greater than or equal to 1800 and the second molecular weight of the polymer is greater than or equal to 16800 will the functional formulation not be metabolized by the kidneys in large quantities.

[0499] It should be noted that, in the various embodiments of this application, although the functional links and the links to be modified are randomly copolymerized in the corresponding segments, those skilled in the art will know that the same technical effect can still be achieved by using block copolymerization.

[0500] In the various embodiments of this application, although the functional links and the links to be modified are only embedded in the hydrophilic or hydrophobic segments, those skilled in the art will know that the functional links and the links to be modified are few in number and contribute little to the structural strength of the overall polymer single chain. Therefore, they can also be embedded in both the hydrophilic and hydrophobic segments.

[0501] In this application, the molar ratio of fluorescent agent to polymer is preferably 1:20 to 1:80, which can effectively avoid the quenching effect of fluorescent agent.

[0502] In this application, the molar ratio of fluorescent agent to polymer is preferably 1:20 to 1:80, and the molar ratio of first reactive group to polymer is preferably 2:1 to 1:3. Therefore, the number of functional and unmodified segments on the polymer single chain is very small, and their impact on the polymer properties and related mechanisms is negligible. More importantly, the hydrophobic groups in the hydrophobic segments exhibit twisting and torsion characteristics in three-dimensional space, while the first reactive group of the unmodified segment and the fluorescent agent in the functional segment only exhibit planar characteristics. According to the applicant's research and analysis, the binding strength of polymer hydration groups is related to the interlacing, entanglement, or interpenetration of hydrophobic segments among a large number of single chains; therefore, the contribution of unmodified and functional segments to the binding strength of polymer hydration groups is very small.

[0503] The applicant discovered in experiments that methyl and ethyl groups are preferably not used as hydrophobic groups because, in other experiments not shown in this application, the applicant found that methyl and ethyl groups, when used as hydrophobic groups, have a possibility of success, but also a possibility of failure, all other parameters being the same. Analysis revealed that this is mainly because when methyl and ethyl groups are used as hydrophobic groups, the hydrophobic chain segments are shorter, and the degree of kinking and interpenetration is lower, resulting in a certain degree of dispersion in the binding strength of the polymer hydrate groups.

[0504] Although the embodiments in this application do not include examples of using IR-820 or other derivatives as fluorescent agents, the properties of IR-820 are very similar to those of IR-783. Therefore, those skilled in the art will know that IR-820 can also be used as a fluorescent agent option.

[0505] In the embodiments of this application, succinimide ester is mainly used as the first reactive group, which has milder reaction conditions, higher reaction efficiency, and makes it easier for fluorescent agents to be modified onto the chain segments to be modified.

[0506] In the embodiments of this application, the aqueous solution is an isotonic solution, which has good biocompatibility, low toxicity, and is suitable for injection into the blood, and has great potential for clinical application.

[0507] The functional formulation of this application has the advantage of rapid hepatic metabolism to reach the bile duct in fluorescence imaging and fluorescence laparoscopic surgical navigation, enabling clear fluorescence imaging of the bile duct. It also has a long retention time in the bile duct, which is highly beneficial for surgical procedures. Compared with existing technologies, it reduces the occurrence of unclear fluorescence imaging of the bile duct without invasiveness, or provides a material basis or preparation method for achieving the above effects. Furthermore, it avoids the pain caused to patients by iatrogenic bile duct injury during surgery. In addition, the functional formulation of this application exhibits minimal individual variability and high clinical applicability. Especially in cases of liver injury and biliary tract disease, it can still be normally metabolized by the liver, showing broad application prospects.

[0508] The description of the above specification and embodiments is used to explain the scope of protection of this application, but does not constitute a limitation on the scope of protection of this application.

Claims

1. A polymer, which is a collection of polymer single chains; characterized in that: The polymer single chain comprises at least one hydrophilic segment and at least one hydrophobic segment of block copolymer; the hydrophilic segment has a brush-like structure and includes multiple hydrophilic segments, which are conjugated with polyethylene glycol; the hydrophobic segment includes multiple hydrophobic segments, which are conjugated with hydrophobic groups. At least a portion of the polymer single chain also includes functional segments embedded in the hydrophilic segment and / or the hydrophobic segment; the functional segment is defined as a segment conjugated with a fluorescent agent or a substance formed after the fluorescent agent has deactivated, and at least a portion of the functional segment conjugated with a fluorescent agent; In the polymer, the average number of repeating units in polyethylene glycol is between 4 and 8; The first molecular weight of the polymer is between 1800 and 7800; wherein, the first molecular weight of the polymer is defined as the average molecular weight of the hydrophobic ends of the polymer single chains in the polymer, and the hydrophobic ends of the polymer single chains are defined as all hydrophobic segments including the same polymer single chain. The second molecular weight of the polymer is between 16,800 and 27,100; wherein, the second molecular weight of the polymer is defined as the sum of the average molecular weight of the hydrophobic ends of the polymer single chains and the average molecular weight of the hydrophilic ends of the polymer single chains, and the hydrophilic ends of the polymer single chains are defined as all hydrophilic segments of the same polymer single chain.

2. The polymer as described in claim 1, characterized in that, In the polymer, the average number of repeating units in polyethylene glycol is between 5 and 8.

3. The polymer as described in claim 1, characterized in that, In the polymer, the molar ratio of the fluorescent agent to the polymer single chain is preferably between 1:20 and 1:

80.

4. The polymer as described in claim 1, characterized in that, In polymers, the polymeric groups used to form polymer single chains are groups suitable for undergoing living radical polymerization reactions.

5. A polymer as described in claim 4, characterized in that, The polymeric groups are acrylate groups and / or methacrylate groups.

6. The polymer as described in claim 1, characterized in that, The hydrophobic group is any one or more of methyl, ethyl, butyl, hexyl, cyclohexyl, dodecyl, and octadecyl.

7. A polymer as claimed in claim 6, characterized in that, The hydrophobic group is preferably one or more selected from butyl, hexyl, cyclohexyl, and dodecyl.

8. The polymer as described in claim 1, characterized in that, The fluorescent agent is any one or more of the following: indocyanine green or its derivatives, boron fluoride pyrrole or its derivatives, IR-783 or its derivatives, IR-820 or its derivatives, FD-1080 or its derivatives, IR-1060 or its derivatives, Cy7 or its derivatives, and Cy5.5 or its derivatives.

9. A polymer as claimed in claim 8, characterized in that, The fluorescent agent is preferably any one or more of IR-783 or its derivatives, IR-820 or its derivatives, and indocyanine green or its derivatives.

10. A polymer as claimed in claim 1, characterized in that, The polymer single chain has a star-shaped topology, with each arm of the star having both hydrophobic and hydrophilic segments. On the same arm, the hydrophilic segments are further away from the center of the star than the hydrophobic segments.

11. A polymer as claimed in claim 1, characterized in that, The polymer single chain has a linear topology.

12. The polymer as claimed in claim 11, characterized in that, The polymer single chain includes a hydrophilic segment and a hydrophobic segment.

13. A polymer as claimed in claim 11, characterized in that, The polymer single chain sequentially includes a first hydrophilic segment, a hydrophobic segment, and a second hydrophilic segment, both of which are hydrophilic segments.

14. A polymer as claimed in claim 11, characterized in that, The polymer single chain sequentially includes a first hydrophobic segment, a hydrophilic segment, and a second hydrophobic segment, both of which are hydrophobic segments.

15. A polymer according to any one of claims 1 to 14, characterized in that, At least a portion of the polymer single chain also includes a segment to be modified, which is embedded in the hydrophilic segment and / or the hydrophobic segment. The segment to be modified is conjugated with a first reactive group, which is used to conjugate the fluorescent agent by substitution or coupling reaction.

16. A polymer as claimed in claim 15, characterized in that, The first reactive group is any one or more of succinimide ester, maleimide ester, azide, alkynyl, amino, mercapto, hydroxyl and aldehyde.

17. A polymer as claimed in claim 16, characterized in that, The first reactive group is preferably a succinimide ester.

18. A polymer as claimed in claim 15, characterized in that, The molar ratio of the first reactive group to the polymer single chain is preferably between 2:1 and 1:

3.

19. A functional formulation, characterized in that, It includes polymer hydrates dispersed in water or an aqueous solution, at least a portion of which comprises the polymer as claimed in any one of claims 1 to 18.

20. A functional formulation as described in claim 19, characterized in that, The polymer hydrates are dispersed in an isotonic solution.

21. A functional formulation as described in claim 20, characterized in that, The isotonic solution is physiological saline, glucose injection, Ringer's solution, balanced salt solution, or balanced buffer solution.

22. A functional formulation as described in claim 20, characterized in that, The average particle size of the polymer hydrates in the functional formulation is between 30 and 400 nanometers.

23. The use of a functional preparation, characterized in that, The functional formulation according to any one of claims 20 to 22 is used to deliver a fluorescent agent into the bile duct via blood to perform fluorescence imaging of the bile duct or to guide fluorescence laparoscopic bile duct surgery by means of fluorescence imaging of the bile duct.

24. The use of a functional formulation as described in claim 23, characterized in that, The functional preparation is administered into the bloodstream via intravenous injection or infusion, and at least a portion of the fluorescent agent reaches the bile duct within 45 minutes after intravenous injection.

25. The use of a functional formulation as described in claim 23, characterized in that, The fluorescent agent has a residence time in the bile duct of 3 hours or more.

26. The use of a functional formulation as described in claim 23, characterized in that, The functional formulation enables clear fluorescence imaging of the bile duct for a time greater than or equal to 2 hours, wherein clear fluorescence imaging of the bile duct refers to the fluorescence signal intensity of the bile duct and / or gallbladder being significantly greater than that of the liver.

27. The use of a functional preparation, characterized in that, The functional formulation according to any one of claims 20 to 22 is used to deliver a fluorescent agent via blood to the digestive tract downstream of the bile duct for fluorescence imaging of the digestive tract downstream of the bile duct, said digestive tract downstream of the bile duct including the duodenum, small intestine and colon.

28. A method for preparing a polymer, used to prepare the polymer as described in any one of claims 1 to 14, characterized in that, The preparation method includes: The polymer is polymerized by a living free radical polymerization reaction; The hydrophilic segment is polymerized from hydrophilic monomers, and the hydrophilic monomers form the hydrophilic chain segment in the hydrophilic segment; The hydrophobic segment is polymerized from a hydrophobic monomer, which forms the hydrophobic linker in the hydrophobic segment; a functional monomer conjugated with a fluorescent agent is inserted into the corresponding segment during the formation of at least one segment, and the functional monomer is inserted to form the functional linker.

29. A method for preparing a polymer as described in claim 28, characterized in that, The functional monomer is prepared by a substitution reaction or a coupling reaction of a first unit and a second unit. The first unit includes a polymeric group and a first reactive group connected to each other, and the second unit includes a fluorescent agent and a second reactive group suitable for coupling or substitution reactions with the first reactive group connected to each other.

30. A method for preparing a polymer, used to prepare the polymer as described in any one of claims 15 to 18, characterized in that, The preparation method includes: The polymer is polymerized by a living free radical polymerization reaction; The hydrophilic segment is polymerized from hydrophilic monomers, and the hydrophilic monomers form the hydrophilic chain segment in the hydrophilic segment; The hydrophobic segment is polymerized from hydrophobic monomers, and the hydrophobic monomers form the hydrophobic links in the hydrophobic segment; The monomer to be modified, which is conjugated with the first reactive group, is inserted into the corresponding chain segment during the formation of at least one chain segment; The monomers to be modified are spliced ​​together to form a pre-modified chain segment; The fluorescent agent is allowed to be probabilistically modified to the intended modified segment under reaction conditions suitable for substitution or coupling reactions to form a functional segment, and the unmodified fluorescent segment becomes the modified segment.

31. A method for preparing a polymer as described in claim 30, characterized in that, The fluorescent agent and the second reactive group are connected to form a second unit. The second unit and the prepared chain segment to be modified are formed into the functional chain segment through a probabilistic substitution reaction or coupling reaction between the second reactive group and the first reactive group.

32. A method for preparing a polymer according to any one of claims 28 to 31, characterized in that: The average number of repeating units of polyethylene glycol in the polymer is controlled by controlling the average number of repeating units of polyethylene glycol in all hydrophilic monomers. The first molecular weight of the polymer is controlled by controlling the degree of polymerization of the hydrophobic monomer; The second molecular weight of the polymer is controlled by separately controlling the degree of polymerization of the hydrophobic monomer and the degree of polymerization of the hydrophilic monomer.

33. The method for preparing a polymer as described in claim 32, characterized in that: The degree of polymerization of hydrophobic monomers is controlled by adjusting the molar ratio of hydrophobic monomers to chain transfer agents or initiators and the conversion rate of the living radical polymerization reaction used to polymerize hydrophobic segments. The degree of polymerization of hydrophilic monomers is controlled by adjusting the molar ratio of hydrophilic monomers to chain transfer agents or initiators and the conversion rate of the living radical polymerization reaction used to polymerize hydrophilic segments.

34. A method for preparing a functional formulation, used to prepare the functional formulation as described in claim 19, characterized in that, The functional formulation is prepared by dispersing the polymer as described in any one of claims 1 to 18 in water or an aqueous solution.

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