Preparation method and application of amphiphilic high-molecular compound capable of being degraded by sound control

Abstract

The invention relates to the technical field of medicines, and discloses a preparation method and application of an amphiphilic high-molecular compound capable of being degraded by sound control. The amphiphilic high-molecular compound provided by the invention has good ultrasonic responsiveness through a specially designed ultrasonic response structure and an ultrasonic fracture structure, and can be decomposed under the ultrasonic action. The compound can be used for effectively wrapping the effective components of the medicine to form a medicine carrying system with ultrasonic responsiveness, so that targeted delivery and controllable release of the medicine are realized.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to a preparation method and application of an amphiphilic polymer compound capable of acoustic-controlled degradation. Background Art

[0002] Chemotherapy drugs have problems such as large toxic and side effects and easy drug resistance, which greatly limit their clinical applications. Some studies have shown that making existing small molecule chemotherapy drugs into specific drug delivery systems, such as nano-delivery systems, can effectively improve the toxic and side effects of chemotherapy drugs and can also improve the targeting and efficacy of drugs.

[0003] In recent years, with the further research on chemotherapy drug delivery systems, researchers have proposed an exogenous stimulus-responsive nano-drug delivery system with better targeting and precise regulation. It refers to a technology that can release drugs directionally through exogenous stimuli, which can not only achieve the targeted accumulation of drugs but also realize the targeted release of drugs through exogenous stimuli, better meeting the needs of targeted drug use, reducing the toxic and side effects of chemotherapy drugs and further improving the efficacy.

[0004] However, there is no relevant report on the exogenous stimulus-responsive nano-drug delivery system that can achieve the directional delivery and release of chemotherapy drugs at present. Summary of the Invention

[0005] The purpose of the present invention is to overcome the problems in the prior art such as the lack of a more effective exogenous stimulus-responsive nano-drug delivery system for the directional delivery and release of chemotherapy drugs, and to provide a preparation method and application of an amphiphilic polymer compound capable of acoustic-controlled degradation. The amphiphilic polymer compound obtained by the method provided by the present invention can self-assemble to form a highly targeted nano-drug delivery system and can well respond to ultrasonic stimulation to release directionally in vivo.

[0006] To achieve the above purpose, on the one hand, the present invention provides a method for preparing an amphiphilic polymer compound capable of acoustic-controlled degradation, and the method includes:

[0007] (1) Providing a compound represented by formula (B):

[0008]

[0009] Wherein, the R group includes a hydroxyl group and / or a conjugated structure;

[0010] (2) Reacting the compound represented by formula (B), the compound represented by formula (A), and a linking skeleton to obtain an amphiphilic polymer compound capable of acoustic-controlled degradation.

[0011]

[0012] Among them, R3 and R4 are each independently selected from alkyl groups having no more than 3 carbon atoms; both R1 and R2 contain hydroxyl groups.

[0013] The second aspect of the present invention provides an intermediate product obtained by the method described in the first aspect.

[0014] The third aspect of the present invention provides an amphiphilic polymer compound that can be degraded by sound control, and the compound is obtained by the method described in the first aspect; or

[0015] The compound includes structural units provided by the compounds shown in formula (A) and formula (B), and a connecting backbone:

[0016]

[0017] Among them, in formula (A), R3 and R4 are each independently selected from alkyl groups having no more than 3 carbon atoms; both R1 and R2 contain hydroxyl groups;

[0018] In formula (B), the R group includes a hydroxyl group and / or a conjugated structure.

[0019] The fourth aspect of the present invention provides the application of the method described in the first aspect, and / or the intermediate product described in the second aspect, and / or the compound described in the third aspect in the preparation of drugs.

[0020] The fifth aspect of the present invention provides an amphiphilic polymer drug delivery system that can be degraded by sound control, and the drug delivery system includes nanoparticles formed by a drug active ingredient and the compound described in the third aspect.

[0021] The sixth aspect of the present invention provides a method for preparing an amphiphilic polymer drug delivery system that can be degraded by sound control, and the method includes: contacting a drug active ingredient with the compound described in the third aspect, and forming nanoparticles through the self-assembly action of the compound.

[0022] Through the above technical solutions, the present invention can at least achieve the following beneficial effects:

[0023] (1) The compound provided by the present invention has a uniquely designed ultrasonic response group and an ultrasonic cleavage group, and can realize the sound-controlled degradation of the polymer drug delivery system, thereby forming an amphiphilic drug molecule with ultrasonic response function. This drug molecule not only has good effects of targeted delivery and aggregation, but also can realize the directional and timed release of the drug through ultrasonic stimulation, making the release of chemotherapy drugs more precisely controllable, improving the chemotherapy effect and reducing side effects.

[0024] (2) The nano-drug delivery system prepared from the compounds provided by the present invention and capable of acoustic-controlled degradation has good stability. The chemotherapeutic drugs loaded therein will not be released without ultrasonic treatment, thereby improving the controllability of chemotherapeutic drugs during tumor treatment and reducing the systemic side effects of chemotherapeutic drugs during use. Description of the Drawings

[0025] Figure 1 It is the 1H NMR spectrum of precursor a obtained in Example 1.

[0026] Figure 2 It is the 1H NMR spectrum of precursor b obtained in Example 1.

[0027] Figure 3 It is the 1H NMR spectrum of precursor c obtained in Example 1.

[0028] Figure 4 It is the 1H NMR spectrum of precursor d obtained in Example 1.

[0029] Figure 5 It is the 1H NMR spectrum of precursor e obtained in Example 1.

[0030] Figure 6 It is the 1H NMR spectrum of precursor f obtained in Example 1.

[0031] Figure 7 It is the 1H NMR spectrum of the acoustic-controlled degradation polymer SDP obtained in Example 1.

[0032] Figure 8 It is the analysis chart of ROS generated by the acoustic-controlled degradation polymer SDP under ultrasonic action measured by an ESR spectrometer in Test Example 1.

[0033] Figure 9 It is the structural formula of the acoustic-controlled degradation polymer SDP obtained in Example 1 and the schematic diagram of the response degradation of SDP under ultrasonic action.

[0034] Figure 10 It is the analysis chart of the degradation of the acoustic-controlled degradation polymer SDP under ultrasonic conditions measured by HPLC in Test Example 2.

[0035] Figure 11 It is the particle size chart and Zeta potential analysis chart of NP-SDP detected by DLS in Test Example 3.

[0036] Figure 12 It is the morphology of the nanoparticles NP-SDP measured by transmission electron microscopy (TEM) in Test Example 4 and the morphology chart of the degradation of NP-SDP under ultrasonic conditions.

[0037] Figure 13It is an analysis diagram of the drug release situation of nanoparticles NP-SDP detected by HPLC under ultrasonic conditions in Test Example 5. Detailed implementation mode

[0038] In the ranges disclosed herein, the endpoints and any values are not limited to the exact ranges or values, and these ranges or values should be understood to include values close to these ranges or values. For numerical ranges, the endpoint values of each range, between the endpoint values of each range and individual point values, and between individual point values can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed herein.

[0039] In the present invention, "acoustic-controlled degradation" refers to the process in which under the action of ultrasonic waves, compound molecules or polymers formed by the compound molecules can be degraded, thereby exposing the drug molecules encapsulated by them and realizing the release of drug molecules. "Amphiphilic polymer compound capable of acoustic-controlled degradation" refers to a compound (polymer) having the above characteristics.

[0040] In the present invention, without special instructions, "DMF" is the abbreviated form of "N,N-dimethylformamide", and they have the same meaning; "EtOH" is the abbreviated form of "ethanol", and they have the same meaning; "DMSO" is the abbreviated form of "dimethyl sulfoxide", and they have the same meaning; "Ola" is the abbreviated form of "Olaparib", and their meanings are the same as those of "olaparib"; "PEG" is the abbreviated form of "polyethylene glycol", and they have the same meaning; the above terms with the same meaning can be used interchangeably.

[0041] In the present invention, without special instructions, "eq" refers to "equivalent", and one equivalent (1eq) is the amount corresponding to a compound gaining / losing one electron.

[0042] In the present invention, the numbers for operations, reagents, etc. (such as "first mixing", "reagent A", "organic solvent A", etc.) are only used for facilitating the distinction of the corresponding operations and reagents in different steps in the description, and have no restrictive effect on the specific reagent selection and operation sequence / manner / conditions, etc. The operations / reagents with different numbers in different steps can be the same or different.

[0043] The currently developed exogenous stimulus-responsive nano-drug delivery systems mainly include two types: photo-responsive and ultrasound-responsive. Among them, the inventors of the present invention found during the research process that although the photo-responsive has good directivity, light is difficult to penetrate the skin, thus being limited to the application of skin surface tumors only. Moreover, long-term laser irradiation at a fixed position is likely to cause skin ulceration, resulting in unnecessary side effects. The action range of ultrasound is relatively wide and it has good penetrability. In addition, long-term ultrasound action will not only cause side effects, but also has certain effects such as promoting blood circulation to remove blood stasis, promoting nerve healing, and relieving pain, thus making the ultrasound-responsive nano-drug delivery system a more ideal system for the directional and controllable release of chemotherapy drugs. Through further research, the inventors ingeniously designed and synthesized precursor molecules with ultrasound-responsive activity respectively, and designed and synthesized a linking backbone molecule that can simultaneously load them through a condensation reaction, achieving the purpose of having both an ultrasound-responsive part and an ultrasound-cleavable part on the same drug molecule. In addition, the inventors also made the polymer have good amphiphilicity by ingenious design, improving the ability to load lipophilic small molecule chemotherapy drugs. The inventors further found through research that by self-assembling the ultrasound-responsive polymer with small molecule chemotherapy drugs to form nanoparticles with certain targeting functions, it is not only convenient for drug administration, but also can utilize the responsiveness of the polymer to achieve the controllable degradation of nanoparticles and the release of chemotherapy drugs, reducing the systemic side effects of the drugs.

[0044] Based on the above findings, in the first aspect of the present invention, a method for preparing an amphiphilic polymer compound that can be degraded by acoustic control is provided, and the method includes:

[0045] (1) providing a compound represented by formula (B):

[0046]

[0047] wherein the R group includes a hydroxyl group and / or a conjugated structure;

[0048] (2) reacting the compound represented by formula (B), the compound represented by formula (A), and a linking backbone to obtain an amphiphilic polymer compound that can be degraded by acoustic control,

[0049]

[0050] wherein R3 and R4 are each independently selected from alkyl groups having no more than 3 carbon atoms; both R1 and R2 contain hydroxyl groups.

[0051] In the amphiphilic polymer compound prepared by the method provided by the present invention and capable of acoustic-controlled degradation, it mainly includes an acoustic-controlled degradation ultrasonic fracture structure and an ultrasonic response structure, as well as a connecting backbone that connects the ultrasonic fracture structure and the ultrasonic response structure to form a polymer. In this compound, the ultrasonic fracture structure and the ultrasonic response structure are respectively shown in formula (A) and formula (B). Those skilled in the art can design the corresponding connecting backbone according to their characteristics. In addition, according to the characteristics and functions of the structures of formula (A) and formula (B), those skilled in the art can also specifically select a suitable compound as the precursor of the amphiphilic polymer compound provided by the present invention and synthesize this compound.

[0052] For example, in the present invention, the main function of the compound shown in formula (A) is to enable the polymer compound to achieve acoustic-controlled degradation. Any linking bond and substituent that can cause it to react and break under ultrasonic conditions with reactive oxygen species (ROS) can be applicable to formula (A). For example, in formula (A), R1 and R2 are each independently selected from an alkyl group having no more than 5 carbon atoms and having at least one hydroxyl group at the end.

[0053] In the method provided by the present invention, step (1) is the process of providing a precursor of the ultrasonic response structure in the compound provided by the present invention (i.e., the compound shown in formula (B)). Those skilled in the art can design the synthesis method of its corresponding precursor according to its characteristics, or obtain this compound precursor through commercial purchase or customization.

[0054] According to a particularly preferred embodiment of the present invention, step (1) includes:

[0055] (1-1) Contacting the compounds shown in formula (4) and formula (5) in the presence of organic solvent A and carrying out a first reaction to obtain a product shown in formula (6). Preferably, the first reaction temperature is 135 - 145 °C.

[0056]

[0057] (1-2) Contacting the compounds shown in formula (6) and formula (7) in the presence of organic solvent B and carrying out a second reaction to obtain a product shown in formula (8). Preferably, the second reaction temperature is 20 - 35 °C.

[0058]

[0059] (1-3) Contacting the compound shown in formula (8) with reagent A in the presence of organic solvent C and carrying out a third reaction to obtain a product shown in formula (9). Preferably, the third reaction temperature is 135 - 145 °C.

[0060]

[0061] (1-4) Contact the compounds with the structures shown in formulas (9) and (10) in the presence of organic solvent D and carry out the fourth reaction to obtain the product with the structure shown in formula (11). Preferably, the temperature of the fourth reaction is 105-115 °C.

[0062]

[0063] (1-5) Contact the compounds with the structures shown in formulas (11) and (12) in the presence of organic solvent E and carry out the fifth reaction to obtain the product with the structure shown in formula (13). Preferably, the temperature of the fifth reaction is 84-86 °C.

[0064]

[0065] (1-6) Contact the compound with the structure shown in formula (13) with reagent B in the presence of organic solvent F and carry out the sixth reaction to obtain the product shown in structural formula (2) (i.e., the precursor of the ultrasonic response structure in the compound). Preferably, the temperature of the sixth reaction is 20-30 °C.

[0066]

[0067] When the above method is used to prepare the precursor of the ultrasonic response structure in the compound provided by the present invention, according to the preferred embodiment of the present invention, in step (1-1), the organic solvent A is selected from N,N-dimethylformamide. Preferably, it is ultra-dry DMF.

[0068] Preferably, relative to 1 mmol of the compound shown in formula (4), the amount of organic solvent A used is 10-20 mL.

[0069] According to the preferred embodiment of the present invention, in step (1-1), relative to 1 mmol of the compound shown in formula (4), the amount of the compound shown in formula (5) used is 0.8-1.5 eq.

[0070] Preferably, step (1-1) further includes adding potassium carbonate. Preferably, relative to 1 mmol of the compound shown in formula (4), the amount of potassium carbonate used is 0.5-1.5 eq.

[0071] According to the preferred embodiment of the present invention, in step (1-1), the process of purifying the reaction product is further included. Preferably, silica gel column chromatography is used for product purification.

[0072] According to the preferred embodiment of the present invention, in step (1-2), the organic solvent B is selected from N,N-dimethylformamide. Preferably, it is ultra-dry DMF.

[0073] Preferably, relative to 1 mmol of the compound represented by formula (6), the amount of organic solvent B used is 1 - 2 mL.

[0074] According to a preferred embodiment of the present invention, in step (1 - 2), relative to 1 mmol of the compound represented by formula (6), the amount of the compound represented by formula (7) used is 1.2 - 1.5 mmol.

[0075] Preferably, step (1 - 2) further includes adding the compound represented by formula (5) and / or potassium carbonate. Preferably, relative to 1 mmol of the compound represented by formula (6), the amount of the compound represented by formula (5) used is 0.8 - 1.5 mmol; preferably, the amount of potassium carbonate used is 0.5 - 1.5 mmol.

[0076] According to a preferred embodiment of the present invention, in step (1 - 2), it further includes a process of purifying the reaction product. Preferably, silica gel column chromatography is used for product purification.

[0077] According to a preferred embodiment of the present invention, in step (1 - 3), the organic solvent C is selected from an aqueous ethanol solution, preferably with a volume ratio of ethanol to water of 3 - 6:1.

[0078] Preferably, relative to 1 mmol of the compound represented by formula (8), the amount of organic solvent C used is 1 - 2 mL.

[0079] According to a preferred embodiment of the present invention, in step (1 - 3), relative to 1 mmol of the compound represented by formula (8), the amount of reagent A used is 4.5 - 5.5 mmol.

[0080] Preferably, in step (1 - 3), reagent A is selected from iron powder.

[0081] Preferably, step (1 - 3) further includes adding ammonium chloride. Preferably, relative to 1 mmol of the compound represented by formula (8), the amount of ammonium chloride used is 0.5 - 1.5 mmol.

[0082] According to a preferred embodiment of the present invention, in step (1 - 3), it further includes a process of purifying the reaction product. Preferably, silica gel column chromatography is used for product purification.

[0083] According to a preferred embodiment of the present invention, in step (1 - 4), the organic solvent D is selected from toluene, preferably super - dry toluene.

[0084] Preferably, relative to 1 mmol of the compound represented by formula (9), the amount of organic solvent D used is 0.4 - 0.6 mL.

[0085] According to a preferred embodiment of the present invention, in step (1-4), the amount of the compound represented by formula (10) is 0.8-0.95 mmol relative to 1 mmol of the compound represented by formula (9).

[0086] Preferably, step (1-4) further comprises adding sodium tert-butoxide, tri(dibenzylideneacetone)dipalladium and tri-tert-butylphosphine. Relative to 1 mmol of the compound represented by formula (9), the amount of sodium tert-butoxide is preferably 0.9-1.5 mmol; the amount of tri(dibenzylideneacetone)dipalladium is preferably 0.001-0.003 mmol; and the amount of tri-tert-butylphosphine is preferably 0.1-0.2 mmol.

[0087] According to a preferred embodiment of the present invention, step (1-4) further comprises a process of purifying the reaction product, preferably using silica gel column chromatography to purify the product.

[0088] According to a preferred embodiment of the present invention, in step (1-5), the organic solvent E is selected from toluene, preferably super dry toluene.

[0089] Preferably, the total amount of the organic solvent E is 8-12 mL relative to 1 mmol of the compound having the structure represented by formula (11).

[0090] According to a preferred embodiment of the present invention, in step (1-5), the amount of the compound represented by formula (12) is 0.3-0.5 mmol relative to 1 mmol of the compound represented by formula (11).

[0091] According to a preferred embodiment of the present invention, step (1-5) further comprises adding sodium tert-butoxide, tri(dibenzylideneacetone)dipalladium and tri-tert-butylphosphine. Relative to 1 mmol of the compound having the structure shown in formula (11), the amount of sodium tert-butoxide is preferably 0.9-1.5 mmol; the amount of tri(dibenzylideneacetone)dipalladium is preferably 0.08-0.15 mmol; and the amount of tri-tert-butylphosphine is preferably 0.5-0.6 mmol.

[0092] According to a preferred embodiment of the present invention, step (1-5) further comprises a process of purifying the reaction product, preferably by using silica gel column chromatography to purify the product.

[0093] According to a preferred embodiment of the present invention, in step (1-6), the organic solvent F is selected from dichloromethane.

[0094] Preferably, the total amount of the organic solvent F used is 10-30 mL relative to 1 mmol of the compound having the structure represented by formula (13).

[0095] According to a preferred embodiment of the present invention, in step (1-6), reagent B is selected from pyridine hydrofluoride and / or tetrabutylammonium fluoride.

[0096] Preferably, relative to 1 mmol of the compound having the structure shown in formula (13), the amount of reagent B used is 1-3 μL.

[0097] According to a preferred embodiment of the present invention, in step (1-5), a process of purifying the reaction product is further included. Preferably, silica gel column chromatography is used for product purification.

[0098] According to a preferred embodiment of the present invention, in step (2), the structural unit of the linking backbone includes the structure shown in formula (C):

[0099]

[0100] Wherein, R5, R6 and R7 are provided by the compounds shown in formula (A) and / or formula (B).

[0101] In the method provided by the present invention, step (2) is a process of reacting the precursor of the ultrasound-responsive structure (the compound shown in formula (B)), the precursor of the ultrasound-cleavable structure (the compound shown in formula (A)) and the precursor of the linking backbone (the compound shown in formula (C)) to obtain the compound provided by the present invention. Those skilled in the art can design the synthesis method of the corresponding precursors by themselves according to the characteristics of the compounds of formula (A), (B) and (C) above, or obtain the compound precursors through commercial purchase or customization, and use these compound precursors to synthesize the compound provided by the present invention through reasonable reactions.

[0102] According to a particularly preferred embodiment of the present invention, in step (2), it includes: in the presence of organic solvent G, contacting the compounds having the structures shown in formula (1) and formula (2), the linking backbone having the structural unit shown in formula (3) and the capping agent, and performing the seventh reaction to obtain the compound having the structure shown in formula (14) (which can also be abbreviated as "SDP" in the present invention). Preferably, the capping agent is polyethylene glycol (such as polyethylene glycol 2000, polyethylene glycol 5000, etc.).

[0103]

[0104] Wherein, n is an integer from 5 to 15, and m is an integer from 50 to 150.

[0105] Preferably, the temperature of the seventh reaction is 30-60 °C.

[0106] Preferably, the organic solvent G is selected from N,N-dimethylformamide. Preferably, it is ultra-dry DMF.

[0107] More preferably, the amount of the organic solvent G is 10 - 20 mL relative to 1 mmol of the compound represented by the formula (2).

[0108] More preferably, the amount of the compound represented by the formula (1) is 0.8 - 1 mmol relative to 1 mmol of the compound represented by the formula (2).

[0109] More preferably, the amount of the linking backbone is 1.1 - 1.2 mmol relative to 1 mmol of the compound represented by the formula (2).

[0110] More preferably, the amount of the capping agent is 0.5 - 0.6 mmol relative to 1 mmol of the compound represented by the formula (3).

[0111] The second aspect of the present invention provides an intermediate product obtained by the method according to the first aspect.

[0112] According to a preferred embodiment of the present invention, wherein the structure of the intermediate product is as shown in the formula (B),

[0113]

[0114] wherein the R group includes a hydroxyl group and / or a conjugated structure.

[0115] Preferably, the structure of the intermediate product is as shown in the formula (2).

[0116]

[0117] The third aspect of the present invention provides an amphiphilic polymer compound that can be degraded by sound control, and the compound is obtained by the method according to the first aspect; or

[0118] The compound includes structural units provided by the compounds represented by the formula (A) and the formula (B), and a linking backbone:

[0119]

[0120] wherein, in the formula (A), R3 and R4 are each independently selected from alkyl groups having no more than 3 carbon atoms (for example, they can be methyl, ethyl, propyl, isopropyl, etc.); both R1 and R2 contain hydroxyl groups (for polymer polymerization);

[0121] In the formula (B), the R group includes a hydroxyl group and / or a conjugated structure.

[0122] According to some preferred embodiments of the present invention, wherein the structural unit of the linking backbone preferably includes the structure shown in the formula (C):

[0123]

[0124] Among them, R5 and, R6 and R7 are provided by the compounds represented by formula (A) and / or formula (B).

[0125] According to a particularly preferred embodiment of the present invention, the structural unit of the compound includes the compounds represented by formula (1) and formula (2), and the compound further includes a connecting backbone as shown in formula (3):

[0126]

[0127] Preferably, the compound further includes a capping agent, and preferably the capping agent is polyethylene glycol (such as polyethylene glycol 2000, polyethylene glycol 5000, etc.). Other commonly used capping agents in the art can also be selected for the compounds provided by the present invention. Those skilled in the art can reasonably design the connecting backbone molecules when using other capping agents according to the characteristics of the compounds provided by the present invention and the characteristics of other capping agents, which will not be elaborated here.

[0128] According to a particularly preferred embodiment of the present invention, the structure of the compound is as shown in formula (14) (this compound can also be simply referred to as "SDP" in the present invention), wherein n is an integer from 5 to 15, and m is an integer from 50 to 150.

[0129]

[0130] The fourth aspect of the present invention provides the application of the method described in the first aspect, and / or the intermediate product described in the second aspect, and / or the compound described in the third aspect in the preparation of drugs.

[0131] The fifth aspect of the present invention provides an acoustically controllable degradable amphiphilic polymer drug delivery system, and the drug delivery system includes nanoparticles formed by a drug active ingredient and the compound described in the third aspect.

[0132] In the drug delivery system provided by the present invention, the drug active ingredient refers to a small molecule compound that exerts a drug effect. Any small molecule drug commonly used in the art can be used as the drug active ingredient in the drug delivery system provided by the present invention.

[0133] Since the above-mentioned acoustically controllable degradable compound provided by the present invention has a good encapsulation effect on the drug active ingredient, and the drug delivery system formed after encapsulation has high stability, it can effectively protect the drug active ingredient without ultrasonic treatment and prevent it from being released during the in vivo transportation process, thereby improving the targeted delivery and release of the drug, effectively reducing drug side effects, and making the drug delivery system provided by the present invention particularly suitable for drugs with requirements for directional delivery and release, such as anti-cancer chemotherapy drugs.

[0134] According to a preferred embodiment of the present invention, the pharmaceutically active ingredient includes an anticancer chemotherapy drug, preferably selected from lipophilic small molecule chemotherapy drugs.

[0135] Preferably, the pharmaceutically active ingredient is selected from Olaparib and / or Niraparib.

[0136] According to a preferred embodiment of the present invention, the average particle size of the nanoparticles is 80 - 120 nm, and the zeta potential is -5 mV to -25 mV.

[0137] Preferably, the average particle size of the nanoparticles is 100 - 120 nm, and the Zeta potential is -15 mV to -25 mV.

[0138] The sixth aspect of the present invention provides a method for preparing a sound - controllable degradable amphiphilic polymer drug - loading system, the method comprising: contacting the pharmaceutically active ingredient with the compound described in the third aspect, and forming nanoparticles through the self - assembly of the compound.

[0139] According to a preferred embodiment of the present invention, the method comprises:

[0140] Dissolving the compound and the pharmaceutically active ingredient in organic solvent I to obtain a blend solution I, mixing the blend solution I with water for the first time, and then removing the organic solvent I therein to obtain nanoparticles.

[0141] Preferably, the organic solvent I is selected from dimethyl sulfoxide and / or tetrahydrofuran.

[0142] Preferably, the pharmaceutically active ingredient includes an anticancer chemotherapy drug, preferably selected from lipophilic small molecule chemotherapy drugs.

[0143] More preferably, the pharmaceutically active ingredient is selected from Olaparib and / or Niraparib.

[0144] Preferably, in the blend solution I, the content of the pharmaceutically active ingredient is 1 - 20 mg / mL, the content of the compound is 20 - 200 mg / mL, and preferably the (mass) content of the compound is 5 - 15 times that of the pharmaceutically active ingredient.

[0145] Preferably, the mode of the first mixing includes: adding the blend solution I dropwise into water under stirring conditions, and continuing to stir for 5 - 10 min after the dropping is completed.

[0146] Preferably, the volume ratio of the blend solution I to water is 1:5 - 15.

[0147] Preferably, the organic solvent I is removed by dialysis.

[0148] The present invention will be described in detail below through examples. It should be understood that the following examples are only used to further explain and illustrate the content of the present invention by way of example, and are not used to limit the present invention.

[0149] In the following examples, unless otherwise specified, the reagents and materials used are all commercially available products purchased from regular chemical / biological reagent / material suppliers, and the reagents are all of analytical grade.

[0150] In the following examples, unless otherwise specified, a silica gel column is used in each step to chromatographically separate the obtained reaction product to purify the product.

[0151] In the following examples, unless otherwise specified, the operations are all carried out at room temperature (25 ± 5 °C).

[0152] Example 1

[0153] This example is used to illustrate the synthesis of the amphiphilic polymer compound capable of acoustic-controlled degradation provided by the present invention.

[0154] Synthesize the amphiphilic polymer compound SDP capable of acoustic-controlled degradation according to the following steps:

[0155] (1) Synthesize the key precursor f (the structure is shown in formula (2)):

[0156] (1-1) Referring to reaction formula (1.1), synthesize precursor a (the structure is shown in formula (6)):

[0157] Dissolve p-nitrophenol (16.7 g, 12 mmol) and potassium carbonate (16.56 g, 12 mmol) in 200 mL of ultra-dry DMF, stir for 10 min, dissolve 3-bromo-1-propanol (13.9 g, 10 mmol) in 50 mL of ultra-dry DMF, add it dropwise to the reaction system, heat to 140 °C and reflux, monitor the reaction progress by thin layer chromatography (TLC) during the reaction, after 6 h, collect the reaction system, evaporate to dryness, and separate by column chromatography to obtain precursor a ( 1 The H NMR analysis results are as Figure 1 shown).

[0158]

[0159] (1-2) Referring to reaction formula (1.2), synthesize precursor b (the structure is shown in formula (8)):

[0160] Precursor a (5 g, 25.38 mmol) was dissolved in 25 mL of ultra-dry DMF, tert-butyldimethylsilyl chloride (4.6 g, 30.52 mmol) was added, and the mixture was stirred at room temperature for 5 min. Imidazole (2.73 g, 40.15 mmol) was added to the reaction system, and the reaction progress was monitored by TLC. The reaction was stopped after 36 h, and an appropriate amount of water (H2O) and ethyl acetate were added for extraction. The organic phase was washed with a saturated NaCl solution, dried over anhydrous Na2SO4, filtered, and the organic phase was spin-dried. Precursor b ( 1 The results of H NMR analysis are as follows Figure 2 shown).

[0161]

[0162] (1-3) Referring to reaction formula (1.3), the precursor c (structure is shown in formula (9)) is synthesized:

[0163] Precursor b (6 g, 19.29 mmol) and NH4Cl (0.62 g, 11.59 mmol) were dissolved in ethanol solution (EtOH:H2O=4:1, 20 mL), iron powder (5.39 g, 96.5 mmol) was added, the temperature was raised to 140°C and refluxed, the solution turned to earth yellow, the reaction progress was monitored by TLC, the reaction was stopped after 2 h, the solid was filtered out, the liquid phase was extracted with ethyl acetate, washed twice with water, the organic phase was collected, dried with Na2SO4 and then spin-dried, and the precursor c ( 1 The results of H NMR analysis are as follows Figure 3 shown).

[0164]

[0165] (1-4) Referring to reaction formula (1.4), the precursor d (structure is shown in formula (11)) was synthesized:

[0166] Under nitrogen protection, 1-(4-bromophenyl)-1,2,2-triphenylethylene (534 mg, 1.3 mmol), sodium tert-butoxide (163 mg, 1.7 mmol), tri(dibenzylideneacetone)dipalladium (4 mg, 0.0042 mmol) were dissolved in 25 mL of anhydrous toluene, and then 10 mL of toluene was used to dissolve precursor c (482 mg, 1.7 mmol) and tri-tert-butylphosphine (4.1 mg, 0.021 mmol), respectively. The dissolved precursor c and tri-tert-butylphosphine were added dropwise to the reaction system, and the temperature was raised to 110° C. and heated to reflux. The reaction should be maintained under anhydrous conditions during the reaction. The reaction progress was monitored by TLC. The reaction was stopped after 24 h, deionized water and dichloromethane were added for extraction, the organic phase was separated and collected, dried with Na2SO4 and then spin-dried, and the precursor d ( 1 The results of H NMR analysis are as follows Figure 4shown).

[0167]

[0168] (1-5) Referring to reaction formula (1.5), the precursor e (structure is shown in formula (13)) was synthesized:

[0169] Under nitrogen protection, the precursor d (1.5 g, 2.24 mmol), 4,7-bis(5-bromothiophene-2-yl)-5,6-dinitro-2,1,3-benzothiadiazole (411 mg, 0.75 mmol), tri(dibenzylideneacetone)dipalladium (204 mg, 0.22 mmol) and sodium tert-butoxide (255.5 mg, 2.66 mmol) were dissolved in 15 mL of anhydrous toluene, and tri-tert-butylphosphine (635 mg, 3.15 mmol) was dissolved in 5 mL of anhydrous toluene. The dissolved tri-tert-butylphosphine was added dropwise to the reaction system, and the temperature was raised to 85° C. and heated to reflux. The reaction should be kept under light-proof and anhydrous conditions. The reaction progress was monitored by TLC. The reaction was stopped after 24 h, deionized water and dichloromethane were added for extraction, the organic phase was separated and collected, dried with Na2SO4 and then spin-dried, and the precursor e ( 1 The results of H NMR analysis are as follows Figure 5 shown).

[0170]

[0171] (1-6) Referring to reaction formula (1.6), the precursor f (structure is shown in formula (2)) is synthesized:

[0172] Precursor e (300 mg, 0.186 mmol) was dissolved in 15 mL of ultra-dry dichloromethane and added to a 50 mL plastic centrifuge tube. Hydrogen fluoride pyridine (300 μL) was slowly added dropwise to the reaction system and stirred at room temperature. The reaction process should be strictly maintained under light-proof and anhydrous conditions. The reaction progress was monitored by TLC. The reaction was stopped after 10 min. Column chromatography silica gel powder was added to the reaction system and then dried. Precursor f ( 1 The results of H NMR analysis are as follows Figure 6 shown).

[0173]

[0174] (2) Referring to reaction formula (1.7), synthesize ultrasound-responsive amphiphilic polymer SDP (structure as shown in formula (14)):

[0175] Under nitrogen protection, 2,2'-(propane-2,2-diylbis(sulfanediyldiyl))bis(ethan-1-ol) (34 mg, 0.17 mmol) and precursor f (100 mg, 0.07 mmol) were dissolved in 10 mL of ultradry DMF. The mixture was stirred for 10 min in the dark. After the system was well mixed, 1,2,4,5-cyclohexanetetracarboxylic dianhydride (60 mg, 0.27 mmol) was dissolved in 5 mL of ultradry DMF and added dropwise to the reaction system. The reaction was continuously stirred at 35 °C for 48 h. Polyethylene glycol 5000 monomethyl ether (240 mg, 0.05 mmol) was added, and the reaction was continuously stirred at 40 °C for 24 h. Finally, SDP was obtained by dialysis and vacuum drying 1 The results of 1H NMR analysis are as Figure 7 shown.

[0176]

[0177] Example 2

[0178] This example is used to illustrate the preparation of the drug-loading system (nanoparticles) provided by the present invention.

[0179] 40 mg of the compound SDP prepared in the example and 5 mg of olaparib were weighed and dissolved in 200 μL of DMSO. After complete dissolution, the solution was slowly added dropwise to 5 mL of ultrapure water and stirred for 10 min. During this period, the solution gradually turned light blue, indicating that the nanoparticles had been successfully prepared. The obtained reaction product was placed in a dialysis bag (MWCO: 3500 Da) for dialysis to remove DMSO. After 48 h, the dialysis solution was obtained, centrifuged, and the supernatant was taken. After lyophilization, the drug-loading system nanoparticles (abbreviated as "NP-SDP") were obtained.

[0180] Test Example 1

[0181] This test example is used to illustrate the ability of the drug-loading system nanoparticles provided by the present invention to generate reactive oxygen species.

[0182] An appropriate amount of NP-SDP prepared in Example 2 was weighed and dissolved in water. The obtained solution was ultrasonically treated for 2 min under the conditions of 1 W / cm 2 , frequency 3 MHz, and duty cycle 50%. The ROS content in the solution was detected by an ESR spectrometer before and after ultrasonic treatment. The results are as Figure 8 shown. It can be seen from the figure that more ROS were generated after NP-SDP was ultrasonically treated.

[0183] The ultrasonic response principle of the polymer SDP is as Figure 9As shown, the compound provided by the present invention contains an ultrasonic response structure (blue part in the figure), an ultrasonic fracture structure (red part in the figure), a connecting skeleton (yellow part in the figure), and a capping agent (green part in the figure). Under the action of ultrasound, the ultrasonic fracture structure breaks, the SDP decomposes, and at the same time ROS is generated. When nanoparticles are prepared using SDP and the active pharmaceutical ingredient (such as olaparib in Example 2), under the action of ultrasonic waves, the SDP in the nanoparticles decomposes, thereby exposing the encapsulated active pharmaceutical ingredient and achieving precise drug release.

[0184] Test Example 2

[0185] This test example is used to illustrate the ultrasonic responsiveness of the compound provided by the present invention.

[0186] Weigh an appropriate amount of SDP prepared in Example 1 and dissolve it in water. The resulting solution is ultrasonically treated for 5 min under the conditions of 1 W / cm 2 , a frequency of 3 MHz, and a duty cycle of 50%. The SDP solution is detected by high performance liquid chromatography (HPLC) before and after ultrasound. The results are as Figure 10 shown. It can be seen from the figure that under the action of ultrasound, SDP significantly degrades, indicating that SDP has good ultrasonic responsiveness.

[0187] Test Example 3

[0188] This test example is used to illustrate the particle size and surface potential characterization results of the drug-loaded system nanoparticles provided by the present invention.

[0189] The particle size and surface potential of the nanoparticles NP-SDP prepared in Example 2 are characterized using a laser particle size analyzer (DLS, Malvern Zetasizer Nano ZS90, Nano ZS, UK).

[0190] The results are shown in detail in Figure 11 . It can be seen from the figure that the average particle size of NP-SDP obtained by DLS detection is 105.5 nm, and the Zeta potential is about -22.7 mV.

[0191] Test Example 4

[0192] This test example is used to illustrate the ultrasonic responsiveness of the drug-loaded system nanoparticles provided by the present invention.

[0193] Weigh an appropriate amount of NP-SDP prepared in Example 2 and dissolve it in water. The resulting solution is ultrasonically treated for 5 min under the conditions of 1 W / cm 2 , a frequency of 3 MHz, and a duty cycle of 50%. The morphology of NP-SDP is detected by transmission electron microscopy (TEM, JEM-2200FS, Hitachi, Japan) before and after ultrasound. The results are asFigure 12 As shown, it can be seen from the figure that after the NP-SDP is ultrasonically treated, the overall structure and morphology are severely damaged, indicating that the NP-SDP exhibits strong ultrasonic responsiveness.

[0194] Test Example 5

[0195] This test example is used to illustrate the drug release effect of the drug-loaded system nanoparticles provided by the present invention.

[0196] Referring to Figure 13 the process in (A), the NP-SDP prepared in Example 2 was dissolved in water, and the obtained solution was ultrasonically treated under the conditions of 1 W / cm 2 , a frequency of 3 MHz, and a duty cycle of 50%, and the treatment times were 0, 30 s, 60 s, and 90 s respectively. The treated solutions were respectively loaded into ultrafiltration tubes and centrifuged to obtain filtrates without nanoparticles and macromolecular compounds. The obtained filtrates were respectively detected by HPLC, and the results are as Figure 13 shown in (B). It can be seen from the figure that olaparib was not found in the filtrate without ultrasonic treatment (NP-SDP + US 0 s), indicating that without ultrasonic treatment, the NP-SDP will not release the drug encapsulated therein. However, when ultrasonic treatment was carried out, olaparib was significantly detected in the filtrate (NP-SDP + US 30 s, NP-SDP + US 60 s, NP-SDP + US 90 s), and with the extension of the ultrasonic time, the release rate of olaparib increased significantly, indicating that the NP-SDP can achieve controllable drug release.

[0197] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited thereto. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, including any other suitable combination of each technical feature. These simple modifications and combinations should also be regarded as the content disclosed by the present invention and fall within the protection scope of the present invention.

Claims

1. A method for preparing an amphiphilic polymer compound capable of being degraded by voice control, characterized in that, The method includes: (1) Providing a compound represented by formula (B): Wherein, the R group includes a hydroxyl group and / or a conjugated structure; (2) Reacting the compound represented by formula (B), the compound represented by formula (A), and a linking backbone to obtain an amphiphilic polymer compound that can be degraded by sound control, Wherein, R3 and R4 are each independently selected from alkyl groups having no more than 3 carbon atoms; both R1 and R2 contain a hydroxyl group.

2. The method according to claim 1, wherein Step (1) includes: (1-1) Contacting the compounds represented by formulas (4) and (5) in the presence of organic solvent A and performing a first reaction to obtain a product represented by formula (6). Preferably, the first reaction temperature is 135 - 145 °C, (1-2) Contacting the compounds represented by formulas (6) and (7) in the presence of organic solvent B and performing a second reaction to obtain a product represented by formula (8). Preferably, the second reaction temperature is 20 - 35 °C, (1-3) Contacting the compound represented by formula (8) with reagent A in the presence of organic solvent C and performing a third reaction to obtain a product represented by formula (9). Preferably, the third reaction temperature is 135 - 145 °C, (1-4) Contacting the compounds represented by formulas (9) and (10) in the presence of organic solvent D and performing a fourth reaction to obtain a product represented by formula (11). Preferably, the fourth reaction temperature is 105 - 115 °C, (1-5) Contacting the compounds represented by formulas (11) and (12) in the presence of organic solvent E and performing a fifth reaction to obtain a product represented by formula (13). Preferably, the fifth reaction temperature is 84 - 86 °C, (1-6) Contacting the compound represented by formula (13) with reagent B in the presence of organic solvent F and performing a sixth reaction to obtain a product represented by structural formula (2). Preferably, the sixth reaction temperature is 20 - 30 °C, 3. The method according to claim 2, wherein In step (1-1), the organic solvent A is selected from N,N-dimethylformamide. Preferably, relative to 1 mmol of the compound represented by formula (4), the amount of organic solvent A used is 10 - 20 mL; And / or, in step (1-1), relative to 1 mmol of the compound represented by formula (4), the amount of the compound represented by formula (5) used is 0.8 - 1.5 eq; Preferably, step (1-1) further includes adding potassium carbonate. Preferably, relative to 1 mmol of the compound represented by formula (4), the amount of potassium carbonate used is 0.5 - 1.5 eq; And / or, in step (1-2), the organic solvent B is selected from N,N-dimethylformamide. Preferably, relative to 1 mmol of the compound represented by formula (6), the amount of organic solvent B used is 1 - 2 mL; And / or, in step (1-2), relative to 1 mmol of the compound represented by formula (6), the amount of the compound represented by formula (7) used is 1.2 - 1.5 mmol; Preferably, step (1-2) further comprises adding a compound represented by formula (5) and / or potassium carbonate, preferably, the amount of the compound represented by formula (5) is 0.8-1.5 mmol relative to 1 mmol of the compound represented by formula (6); preferably, the amount of potassium carbonate is 0.5-1.5 mmol; And / or, in step (1-3), the organic solvent C is selected from an ethanol-water solution, preferably wherein the volume ratio of ethanol to water is 3-6:1, and more preferably, the amount of the organic solvent C is 1-2 mL relative to 1 mmol of the compound represented by formula (8); and / or, in step (1-3), the amount of reagent A used is 4.5-5.5 mmol relative to 1 mmol of the compound represented by formula (8); Preferably, in step (1-3), reagent A is selected from iron powder; Preferably, step (1-3) further comprises adding ammonium chloride, preferably, the amount of ammonium chloride is 0.5-1.5 mmol relative to 1 mmol of the compound represented by formula (8); And / or, in step (1-4), the organic solvent D is selected from toluene, and preferably the amount of the organic solvent D is 0.4-0.6 mL relative to 1 mmol of the compound represented by formula (9); And / or, in step (1-4), the amount of the compound represented by formula (10) is 0.8-0.95 mmol relative to 1 mmol of the compound represented by formula (9); Preferably, step (1-4) further comprises adding sodium tert-butoxide, tri(dibenzylideneacetone)dipalladium and tri-tert-butylphosphine, and relative to 1 mmol of the compound represented by formula (9), the amount of sodium tert-butoxide is preferably 0.9-1.5 mmol; the amount of tri(dibenzylideneacetone)dipalladium is preferably 0.001-0.003 mmol; the amount of tri-tert-butylphosphine is preferably 0.1-0.2 mmol; And / or, in step (1-5), the organic solvent E is selected from toluene, and preferably the total amount of the organic solvent E is 8-12 mL relative to 1 mmol of the compound represented by the structure of formula (11); And / or, in step (1-5), the amount of the compound represented by the structure of formula (12) is 0.3-0.5 mmol relative to 1 mmol of the compound represented by the structure of formula (11); And / or, step (1-5) further comprises adding sodium tert-butoxide, tri(dibenzylideneacetone)dipalladium and tri-tert-butylphosphine, wherein the amount of sodium tert-butoxide is preferably 0.9-1.5 mmol relative to 1 mmol of the compound having the structure shown in formula (11); the amount of tri(dibenzylideneacetone)dipalladium is preferably 0.08-0.15 mmol; and the amount of tri-tert-butylphosphine is preferably 0.5-0.6 mmol; And / or, in step (1-6), the organic solvent F is selected from dichloromethane, and preferably the total amount of the organic solvent F is 10-30 mL relative to 1 mmol of the compound represented by the structure of formula (13); And / or, in steps (1-6), reagent B is selected from pyridine hydrofluoride and / or tetrabutylammonium fluoride, and preferably, relative to 1 mg of the compound of formula (13), the dosage of reagent B is 1-3 μL.

4. The method according to claim 1, wherein, In step (2), the structural unit of the linking backbone includes the structure shown in formula (C): Wherein, R5, R6 and R7 are provided by the compound shown in formula (A) and / or formula (B); Preferably, step (2) includes: in the presence of organic solvent G, contacting the compounds of formula (1) and formula (2) and the linking backbone precursor and capping agent of the structural unit shown in formula (3) and performing the seventh reaction to obtain the compound of formula (14), preferably the capping agent is polyethylene glycol, and preferably the temperature of the seventh reaction is 30-60 °C; Wherein, n is an integer from 5 to 15, and m is an integer from 50 to 150.

5. The intermediate product prepared by the method according to any one of claims 1-4, preferably the structure of the intermediate product is as shown in formula (B), Among them, The R group includes a hydroxyl group and / or a conjugated structure; Preferably, the structure of the intermediate product is as shown in formula (2), 6. An amphiphilic polymer compound capable of being degraded by voice control, characterized in that, The compound is prepared by the method according to any one of claims 1-4; or The compound includes the structural units provided by the compounds shown in formula (A) and formula (B), and a linking backbone: Wherein, in formula (A), R3 and R4 each independently selected from alkyl groups with no more than 3 carbon atoms; both R1 and R2 contain hydroxyl groups; In formula (B), the R group includes a hydroxyl group and / or a conjugated structure.

7. The compound according to claim 6, wherein In formula (A), R1 and R2 each independently selected from structures with no more than 5 carbon atoms and at least one hydroxyl group at the end.

8. The compound according to claim 6 or 7, wherein The structural unit of the linking backbone includes the structure shown in formula (C): Wherein, R5, R6 and R7 are provided by the compound shown in formula (A) and / or formula (B).

9. The compound according to any one of claims 6 - 8, wherein, The compound includes the structural units provided by the compounds shown in formula (1) and formula (2), and the compound also includes a linking backbone of the structural unit shown in formula (3): Preferably, the compound also includes a capping agent, preferably the capping agent is polyethylene glycol; More preferably, the compound is as shown in formula (14), wherein, n is an integer from 5 to 15, and m is an integer from 50 to 150, 10. Use of the method according to any one of claims 1-4, and / or, the intermediate product according to claim 5, and / or, the compound according to any one of claims 6-9 in the preparation of drugs.

11. An acoustically controllable biodegradable amphiphilic polymer drug delivery system, characterized in that, The drug delivery system includes nanoparticles formed by a drug active ingredient and the compound according to any one of claims 6-9.

12. The drug-loading system according to claim 11, wherein, The drug active ingredient includes an anticancer chemotherapy drug, preferably selected from lipophilic small molecule chemotherapy drugs; And / or, the average particle size of the nanoparticles is 80-120 nm, and the zeta potential is -5 mV to -25 mV.

13. A method for preparing an amphiphilic polymer drug delivery system that can be degraded by voice control, characterized in that, The method includes: contacting a drug active ingredient and the compound according to any one of claims 6-9, and forming nanoparticles through the self-assembly of the compound.

14. The method according to claim 13, wherein, The method includes: Dissolve the compound and the pharmaceutically active ingredient in organic solvent I to obtain a blend solution I, and perform a first mixing of the blend solution I with water, and then remove the organic solvent I therein to obtain nanoparticles; Preferably, the organic solvent I is selected from dimethyl sulfoxide and / or tetrahydrofuran; Preferably, the pharmaceutically active ingredient includes an anticancer chemotherapeutic drug, preferably selected from lipophilic small molecule chemotherapeutic drugs; Preferably, in the blend solution I, the content of the pharmaceutically active ingredient is 1-20 mg / mL, and the content of the compound is 20-200 mg / mL. Preferably, the content of the compound is 5-15 times that of the pharmaceutically active ingredient; Preferably, the first mixing method includes: adding the blend solution I dropwise into water under stirring conditions, and continuing to stir for 5-10 min after the addition is completed; Preferably, the volume ratio of the blend solution I to water is 1:5-15; Preferably, the organic solvent I is removed by dialysis.

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