A polyamino acid polymer material, preparation method, application, performance evaluation system and method

Abstract

The present invention provides a polyamino acid polymer material having the following chemical formula: R1 is a hydrophilic amino acid repeating segment or a combination of hydrophilic and hydrophobic amino acid repeating segments; R2 is a linker for connecting the hydrophilic group and the hydrophobic group; R3 is a transitional moiety for reinforcing the connection of the hydrophobic group and preventing substitution reactions; the six-membered ring in the formula is selected from at least one of a benzene ring, cyclohexane, heterocyclohexane, cyclohexene, heterocyclohexene, pyridine, or pyran; and n and m are natural numbers. The present invention uses a polyamino acid polymer material instead of PEG lipids, effectively avoiding the effects of pre-existing anti-PEG antibodies, exhibiting good biodegradability, effectively improving the physical dispersion stability of nanoparticles, and increasing the long-term circulation of nanoparticles in the body. The four-component formula of traditional lipid nanoparticles can be reduced to a three-component formula, resulting in excellent stability of the prepared liposomes and nanocrystal preparations over a certain period of time while retaining the inherent drug-loading advantages of liposomes.

Description

Technical Field

[0001] The present invention relates to a drug encapsulation and solubilization technology using liposomes as raw materials, and in particular to a polymer material used for drug loading and volume expansion. Background Art

[0002] Liposomes are bilayer lipid vesicles containing drugs encapsulated in lipid materials. They are primarily used in drug delivery systems to enhance drug efficacy and minimize toxic side effects. Their unique characteristics include targeting and lymphatic targeting, prolonged drug circulation, and improved stability. Cationic liposomes, microvesicles composed of cationic lipids, are particularly widely used in gene therapy and mRNA therapy. Under acidic conditions, ionizable lipids such as ALC-0315, MC3, and SM-102 undergo protonation to form cationic lipids, which bind to negatively charged nucleic acids through electrostatic interactions. These lipids, when mixed with an aqueous solution containing nucleic acids, precipitate and self-assemble into nucleic acid-loaded nanoparticles. Cationic liposomes can also be used as highly efficient delivery vehicles for antigens, peptides, or small molecule drugs, but they can have certain toxic side effects.

[0003] In addition to liposomes, recent advances in new drug formulation technologies have led to the use of drug-loaded nanoparticles and drug nanocrystals. These formulations primarily address the low bioavailability and poor absorption of poorly soluble drugs. However, due to the varying physicochemical properties of poorly soluble drugs, there is currently a lack of high-performance pharmaceutical polymer excipients specifically designed to enhance the solubility and bioavailability of these drugs. Regarding drug nanocrystals, the commonly used pharmaceutical excipient, polyethylene glycol 1000 succinate (TPGS), has an amphiphilic structure and exhibits certain emulsifying and solubilizing properties, providing excellent stabilization and dispersion of drug-loaded nanoparticles or drug nanocrystals. However, it has the following disadvantages: 1) TPGS exhibits physiological activities, such as P-glycoprotein inhibition, which may alter the in vivo behavior of certain drugs and produce side effects; 2) PEG materials have inherent immunogenicity; and 3) the fixed length of the PEG block limits its effectiveness in solubilization and nanodispersion. Liposomes' surface charge, particle size, and other physicochemical properties influence the formation of their protein corona. For example, cationic liposomes tend to adsorb plasma proteins with an isoelectric point (PI) less than 5.5, while anionic liposomes interact more readily with plasma proteins with a PI greater than 5.5. Larger liposomes have greater surface curvature and surface area, enabling greater plasma protein adsorption. Liposome particle size influences blood circulation time and tissue distribution. When liposome particle size exceeds the endothelial space, tissue penetration is reduced.

[0004] Therefore, it is of great practical significance to prepare a new type of liposome to solve the problems of solubilization, emulsification, dispersion, and stabilization of poorly soluble drugs. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a polyamino acid polymer material, a preparation method and an application thereof, aiming to solve the problems existing in the background technology.

[0006] The technical solution adopted by the present invention to solve the above technical problems is:

[0007] In a first aspect of the present invention, the present invention provides a polyamino acid polymer material, the chemical formula of which is shown below:

[0008] (Formula I)

[0009] wherein R1 is a hydrophilic amino acid repeating segment or a combination of hydrophilic and hydrophobic amino acid repeating segments;

[0010] R2 is a linker used to connect the hydrophilic group and the hydrophobic group;

[0011] R3 is a transition group used to strengthen the connection of the hydrophobic group and prevent the occurrence of substitution reaction;

[0012] The six-membered ring in the formula is selected from at least one of a benzene ring, a cyclohexane, a heterocyclohexane, a cyclohexene, a heterocyclohexene, a pyridine or a pyran;

[0013] n and m are natural numbers.

[0014] Furthermore, R1 is selected from at least one of structural formula II, formula III or formula IV:

[0015] (Formula II); (Formula III); (Formula IV);

[0016] Among them, R 21 and R 22 Independently selected from at least one of hydrogen, alkyl, heteroalkyl, aryl, alkenyl, alkynyl, halogen or silicon.

[0017] Furthermore, R2 is selected from at least one of structural formulas V to XI:

[0018] (Formula V); (Formula VI);

[0019] (Formula VII); (Formula VIII);

[0020] (Formula VIIII); (Formula X);

[0021] (Formula XI);

[0022] Among them, R 31 and R 32 Independently selected from at least one of hydrogen, alkyl, heteroalkyl, aryl, alkenyl, alkynyl, halogen or silicon.

[0023] Furthermore, R4, R5, R7, R8, R9, R 10 、R 11 R3 is selected from at least one of hydrogen, sulfur, selenium, alkyl, heteroalkyl, or silicon.

[0024] Furthermore, R6 is selected from at least one of the following structural formulas;

[0025] (Formula XII);

[0026] (Formula XIII);

[0027] (Formula XIIII);

[0028] (Formula XV).

[0029] Furthermore, the auxiliary material of the polyamino acid polymer material includes at least one selected from the group consisting of organic solvents DMSO, DMF, DMAc, DCM, ether, methyl tert-ether, isopropyl ether, n-heptane, n-hexane, and petroleum ether. The auxiliary material referred to in the present invention is a substance that provides reaction conditions such as possibility and convenience for a chemical reaction, such as a solvent or a catalyst.

[0030] The synthetic route of the polyamino acid polymer material is as follows:

[0031] .

[0032] On this basis, the present invention provides the use of the polyamino acid polymer material prepared according to the above method in the preparation of medicines.

[0033] On this basis, the present invention also provides the use of the polyamino acid polymer material prepared according to the above method in dissolution or emulsification.

[0034] On this basis, the present invention also provides the use of the polyamino acid polymer material prepared according to the above method as a drug delivery carrier.

[0035] On this basis, the present invention also provides the application of the polyamino acid polymer material prepared according to the above method in terms of heat resistance.

[0036] On this basis, the present invention also provides the application of the polyamino acid polymer material prepared according to the above method in terms of sterilization.

[0037] In a first aspect, the present invention provides a system and method for evaluating the performance of the above-mentioned polyamino acid polymer material.

[0038] The system specifically includes:

[0039] The SMILES processing module is configured to generate an initial SMILES representation through a molecular structure conversion method;

[0040] The ChemBERTa encoding module is configured to encode the processed SMILES representation into ChemBERTa to generate a molecular embedding vector suitable for multi-target regression prediction;

[0041] The model building and training module is configured to build a Transformer encoder architecture and is pre-trained on a chemical dataset to optimize semantic representation capabilities.

[0042] Evaluation module: It is configured to perform regression prediction through a multi-target regression model and use weighted mean squared error loss to supervise the regression prediction results.

[0043] The specific evaluation method of the system is:

[0044] (1) SMILES processing: Generate the initial SMILES representation through molecular structure conversion method;

[0045] (2) ChemBERTa encoding: The processed SMILES representation is ChemBERTa encoded to generate a molecular embedding vector suitable for multi-target regression prediction (such as CMC, Z-average, PDI);

[0046] (3) Model construction and training: The backbone network ChemBERTa model structure adopts the Transformer encoder architecture inherited from BERT and is pre-trained on chemical datasets (such as PubChem SMILES) to optimize the representation of chemical semantics;

[0047] (4) Perform regression prediction through a multi-target regression model and use the weighted mean square error (MWSE) loss to supervise the regression prediction results.

[0048] The beneficial effects of the present invention are:

[0049] (1) The present invention uses polyamino acid polymer materials instead of PEG lipids, which can effectively circumvent the influence of pre-existing anti-PEG antibodies and fundamentally solve the problem. The polyamino acid polymer material contains both polysarcosine and liposomes, has good biodegradability, effectively improves the physical dispersion stability of nanoparticles, increases the long circulation of nanoparticles in the body, and can reduce the four components of traditional lipid nanoparticles to a three-component formula. This makes the prepared liposomes and nanocrystal preparations have excellent stability for a certain period of time, greatly retaining the drug-carrying advantages of liposomes themselves.

[0050] (2) The product synthesized by the present invention is an amphiphilic block copolymer, namely, the lipophilic segment present in liposomes and the hydrophilic segment of polysarcosine with different polymerization degrees. The amphiphilic structure enables it to have the common properties of common amphiphilic materials such as solubilization, emulsification, drug-loaded nanoparticle stabilizer, drug nanocrystal dispersion stabilizer, decontamination, and wetting.

[0051] (3) Experiments have shown that the product of the present invention exhibits superior effects in terms of emulsification performance, solubility, dispersibility and stability in drug nanocrystals and nucleic acid lipid nanoparticles.

[0052] (4) The present invention also provides a set of prediction systems and methods to make up for the limitations of experiments, which can better infer the functions and effects of compounds, provide valuable references and directions for subsequent research, and greatly reduce the waste of time and resources in the research and development process. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 These are photos of nanosuspensions of the poorly soluble drug paclitaxel (PTX) (TPGS-PTX, CPSS1-PTX, CPSS2-PTX) prepared using TPGS in the prior art and the products CPSS1 and CPSS2 of the present invention, respectively, in Example 3 of the present invention.

[0054] Figure 2 3. These are photos of nanosuspensions of the poorly soluble drug sirolimus (SIRO) prepared using TPGS in the prior art and the products CPSS1 and CPSS2 of the present invention, respectively (TPGS-SIRO, CPSS1-SIRO, CPSS2-SIRO) in Example 3 of the present invention.

[0055] Figure 3 This is a photo of the bottom precipitate of paclitaxel nanocrystals (CPSS1-PTX) prepared using the product CPSS1 in Example 3 of the present invention.

[0056] Figure 4This is a photo of the bottom precipitate of paclitaxel nanocrystals (TPGS-PTX) prepared using TPGS in the prior art in Example 3 of the present invention.

[0057] Figure 5 3 is a graph showing the particle size and distribution of the TPGS-PTX nanocrystal suspension in Example 3 of the present invention.

[0058] Figure 6 3 is a graph showing the particle size and distribution of the CPSS1-PTX nanocrystal suspension in Example 3 of the present invention.

[0059] Figure 7 3 is a graph showing the particle size and distribution of the CPSS2-PTX nanocrystal suspension in Example 3 of the present invention.

[0060] Figure 8 3 is a graph showing the particle size and distribution of the TPGS-SIRO nanocrystal suspension in Example 3 of the present invention.

[0061] Figure 9 3 is a graph showing the particle size and distribution of the CPSS1-SIRO nanocrystal suspension in Example 3 of the present invention.

[0062] Figure 10 3 is a graph showing the particle size and distribution of the CPSS2-SIRO nanocrystal suspension in Example 3 of the present invention.

[0063] Figure 11 This is a fluorescence emission spectrum diagram of the critical micelle concentration determination of the product CPSS3 in Example 4 of the present invention.

[0064] Figure 12 3 is a graph showing the changes in particle size and PDI of TPGS-PTX, CPSS1-PTX, and CPSS2-PTX nanocrystal suspensions stored at 4°C for 50 days in Example 3 of the present invention.

[0065] Figure 13 3 is a graph showing the changes in particle size and PDI of TPGS-SIRO, CPSS1-SIRO, and CPSS2-SIRO nanocrystal suspensions stored at 4°C for 50 days in Example 3 of the present invention.

[0066] Figure 14 1 is a comparison chart of the solubility experimental results of PTX and SIRO in water, TPGS, CPSS1 and CPSS2, respectively, in Example 6 of the present invention. DETAILED DESCRIPTION

[0067] In order to enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention is described in detail, clearly, and completely in the following embodiments in conjunction with the accompanying drawings. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Moreover, based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts, any modifications, equivalent replacements, improvements, etc., should be included in the scope of protection of the present invention.

[0068] To help those skilled in the art better understand the technical solutions of the present invention, we first explain some of the technical terms of the present invention. In the context of the present invention, these explanations are primarily intended to help those skilled in the art understand and implement the technical solutions of the present invention, and are not intended to limit the present invention. Furthermore, other disclosures outside of the present invention may provide different technical interpretations of these technical terms. To avoid technical ambiguity, these explanations should be considered in conjunction with the present invention when understanding the present invention.

[0069] Pharmaceutical excipients

[0070] The pharmaceutical excipients referred to in the present invention refer to excipients and additives used in the production of drugs or preparation of prescriptions. They are substances other than active drugs that have been reasonably evaluated in terms of safety and are included in pharmaceutical preparations.

[0071] Solubilization and solubilization

[0072] During the development of pharmaceutical preparations, many poorly soluble drugs such as volatile oils, fat-soluble vitamins, and steroid hormones have very low solubility in water and cannot reach the concentration required for treatment. In this case, surfactants are often added to increase the solubility of the drug in water. This surfactant that exerts a solubilizing effect is called a solubilizer.

[0073] Liposomes

[0074] When amphiphilic molecules such as phospholipids are dispersed in the aqueous phase, the hydrophobic tails of the molecules tend to aggregate together and avoid the aqueous phase, while the hydrophilic heads are exposed to the aqueous phase, forming closed vesicles with a bilayer structure. A variety of drugs can be encapsulated in the aqueous phase and the bilayer membrane within the vesicles, similar to the structure of ultramicrocapsules. This ultramicrospherical drug-loaded preparation made by encapsulating drugs between lipid bilayer films is called liposomes.

[0075] Cholesterol

[0076] Cholesterol is a neutral lipid and an amphiphilic molecule, but it is more lipophilic than hydrophilic. Because cholesterol has a high polymerization energy, it is difficult to bind to proteins. Instead, it primarily binds to phospholipids, preventing them from aggregating into crystalline structures. Therefore, cholesterol is used as one embodiment of liposomes in this invention.

[0077] Nanocrystals

[0078] Nanocrystals, also known as nanosuspensions, are pure drug submicron colloidal dispersions with a particle size of less than 1 μm formed by dispersing drug particles in a liquid medium using surfactants or polymers as stabilizers.

[0079] Surfactant

[0080] Surfactant refers to a substance that can significantly reduce the surface tension of a liquid. It is called the surfactant of the liquid. Its structure contains polar hydrophilic groups and non-polar hydrophobic groups.

[0081] emulsions

[0082] An emulsion is a heterogeneous liquid dispersion system formed by mixing two immiscible liquids, in which one liquid is dispersed in the other liquid in the form of droplets.

[0083] Critical micelle concentration (CMC)

[0084] The critical micelle concentration (CMC) is a characteristic property of amphiphilic polymer materials. When an amphiphilic polymer reaches a certain concentration in water, further increasing the concentration will result in the formation of micelles. The CMC is the minimum concentration at which surfactant molecules associate to form micelles in a solution. When a surfactant reaches a certain concentration in an aqueous solution, the solubility of some water-insoluble or slightly soluble drugs in the resulting micellar solution can significantly increase, resulting in a transparent micellar solution. This concentration is called the critical micelle concentration. The mechanism of this formation is that surfactant molecules typically possess an amphiphilic structure—one end is hydrophilic and the other is hydrophobic. At low concentrations, surfactant molecules exist in water as single molecules. As concentration increases, the molecules accumulate on the water surface, forming a monolayer. When surface adsorption reaches saturation, excess molecules aggregate to form micelles. Within the micelles, the hydrophobic groups aggregate, while the hydrophilic groups face outward, contacting the water, thereby reducing the surface tension of the water. In the present invention, the critical micelle concentration can be used to verify the solubility properties of polymer materials.

[0085] Accelerated blood clearing (ABC)

[0086] Accelerated blood clearance refers to the binding of anti-PEG antibodies to the surface of PEGylated nanodrugs, which causes complement activation and greatly accelerates the clearance rate of the drug in the body.

[0087] Example 1, a polyamino acid polymer material.

[0088] This embodiment provides a polyamino acid polymer material, the chemical structure of which is shown in Formula I.

[0089] (Formula I)

[0090] in,

[0091] R1 is a hydrophilic amino acid repeating segment or a combination of hydrophilic and hydrophobic amino acid repeating segments; R1 is selected from at least one of structural formula II, formula III or formula IV:

[0092] (Formula II); (Formula III); (Formula IV);

[0093] R 21 and R 22 Independently selected from at least one of hydrogen, alkyl, heteroalkyl, aryl, alkenyl, alkynyl, halogen or silicon.

[0094] R2 is a linker used to connect the hydrophilic group and the hydrophobic group;

[0095] R2 is selected from at least one of the structural formulas V to XI:

[0096] (Formula V); (Formula VI);

[0097] (Formula VII); (Formula VIII);

[0098] (Formula VIIII); (Formula X);

[0099] (Formula XI);

[0100] Among them, R 31 and R 32 Independently selected from at least one of hydrogen, alkyl, heteroalkyl, aryl, alkenyl, alkynyl, halogen or silicon.

[0101] R3 is a transition group used to strengthen the connection of the hydrophobic group and prevent the occurrence of substitution reaction; R3 is selected from at least one of oxygen, sulfur, selenium, alkyl, heteroalkyl or silicon.

[0102] R4, R5, R7, R8, R8, R 10 、R 11 Each of the following is independently selected from at least one of hydrogen, alkyl, heteroalkyl, aryl, alkenyl, alkynyl, halogen or silicon.

[0103] R6 is selected from at least one of the following structural formulas;

[0104] (Formula XII);

[0105] (Formula XIII);

[0106] (Formula XIIII);

[0107] (Formula XV).

[0108] The six-membered ring in the formula is selected from at least one of a benzene ring, a cyclohexane, a heterocyclohexane, a cyclohexene, a heterocyclohexene, a pyridine or a pyran;

[0109] n and m are natural numbers.

[0110] The auxiliary materials of the polyamino acid polymer material include at least one selected from the group consisting of organic solvents DMSO, DMF, DMAc, DCM, ether, methyl tert-ether, isopropyl ether, n-heptane, n-hexane, and petroleum ether. The auxiliary materials referred to in the present invention are substances that provide reaction conditions such as possibility and convenience for chemical reactions, such as solvents and catalysts.

[0111] The synthetic route of the polyamino acid polymer material is as follows:

[0112] .

[0113] Example 2, a method for preparing a polyamino acid polymer material.

[0114] This embodiment provides a method for preparing a polyamino acid polymer material. According to the preparation method of this embodiment, the polyamino acid polymer material described in Example 1, namely CPSS, can be prepared.

[0115]

[0116] The preparation process is as follows:

[0117] 1. Preparation of CPSS1 (i.e. n=14):

[0118] (1) Synthesis of intermediate pSar14S:

[0119] (1.1) Dissolution: Add 5.83 g (28 mmol, 14 eq.) of N-phenyloxycarbonylsarcosine (SarNPC) to a reaction flask. Weigh 1.29 g (10 mmol, 5 eq.) of N,N-diisopropylethylamine (DIPEA) and add 30 mL of dimethylacetamide (DMAc) to the reaction flask. Stir until dissolved.

[0120] (1.2) Polymerization: Weigh 174 mg (2 mmol, 1 eq.) of neopentylamine into the above reaction flask and carry out polymerization at 60°C for 24 h.

[0121] (1.3) Amidation: Weigh 0.30 g (3 mmol, 1.5 eq.) of succinic anhydride and add it to the above reaction system and react at room temperature for 24 h.

[0122] (1.4) Purification: Add an appropriate amount of diethyl ether to the reaction solution and centrifuge for sedimentation. Repeat this process three times. After vacuum drying, 2.32 g of pSar14S as a white solid was obtained in a yield of 98%.

[0123] (2) Synthetic product CPSS1:

[0124] (2.1) Esterification: Weigh 0.59 g (0.5 mmol, 1 eq.) of pSar14S, 0.39 g (1 mmol, 2 eq.) of cholesterol, 124 mg (0.6 mmol, 1.2 eq.) of dicyclohexylcarbodiimide (DCC), and 15.3 mg (0.125 mmol, 0.25 eq.) of 4-dimethylaminopyridine (DMAP) and add all of them to a reaction flask. Then, add 6 mL of dichloromethane (DCM), stir at room temperature for 24 h, and filter to obtain the filtrate.

[0125] (2.2) Purification: Add an appropriate amount of diethyl ether to the filtrate and centrifuge for three times. After vacuum drying, 705.7 mg of CPSS1 was obtained as a white solid with a yield of 91%.

[0126] 2. Preparation of CPSS2 (i.e. n=28):

[0127] (1) Synthesis of intermediate pSar28S:

[0128] (1.1) Dissolution: Add 5.83 g (28 mmol, 28 eq.) of N-phenyloxycarbonylsarcosine (SarNPC) to a reaction flask, weigh 0.65 g (5 mmol, 5 eq.) of N,N-diisopropylethylamine (DIPEA), and add 30 mL of dimethylacetamide (DMAc) to the reaction flask, and stir to dissolve.

[0129] (1.2) Polymerization: Weigh 87.2 mg (1 mmol, 1 eq.) of neopentylamine into the above reaction flask and carry out polymerization at 60 °C for 24 h.

[0130] (1.3) Amidation: Weigh 0.50 g (5 mmol, 5 eq.) of succinic anhydride and add it to the above reaction system and react at room temperature for 24 h.

[0131] (1.4) Purification: Add an appropriate amount of diethyl ether to the reaction solution and centrifuge for sedimentation. Repeat this process three times. After vacuum drying, 2.13 g of pSar28S as a white solid was obtained in a 98% yield.

[0132] (2) Synthetic product CPSS2:

[0133] (2.1) Esterification: Weigh 1.09 g (0.5 mmol, 1 eq.) of pSar28S, 0.29 g (0.75 mmol, 1.5 eq.) of cholesterol, 124 mg (0.6 mmol, 1.2 eq.) of dicyclohexylcarbodiimide (DCC), and 30.5 mg (0.25 mmol, 0.5 eq.) of 4-dimethylaminopyridine (DMAP) and add all of them to a reaction flask. Then, add 10 mL of dichloromethane (DCM), stir at room temperature for 24 h, and filter to obtain the filtrate.

[0134] (2.2) Purification: Add an appropriate amount of diethyl ether to the filtrate and centrifuge for three times. After vacuum drying, 1.21 g of white solid CPSS2 was obtained with a yield of 95%.

[0135] 3. Preparation of CPSS3 (i.e. n=25):

[0136] (1) Synthesis of intermediate pSar25S:

[0137] (1.1) Dissolution: Add 5.23 g (25 mmol, 25 eq.) of N-phenyloxycarbonylsarcosine (SarNPC) to a reaction flask, weigh 0.65 g (5 mmol, 5 eq.) of N,N-diisopropylethylamine (DIPEA), and add 30 mL of dimethylacetamide (DMAc) to the reaction flask, and stir to dissolve.

[0138] (1.2) Polymerization: Weigh 87.2 mg (1 mmol, 1 eq.) of neopentylamine into the above reaction flask and carry out polymerization at 60 °C for 24 h.

[0139] (1.3) Amidation: Weigh 1 g (10 mmol, 10 eq.) of succinic anhydride and add it to the above reaction system and react at room temperature for 24 h.

[0140] (1.4) Purification: Add an appropriate amount of diethyl ether to the reaction solution and centrifuge for sedimentation. Repeat this process three times. After vacuum drying, 1.94 g of pSar25S as a white solid was obtained in a 99% yield.

[0141] (2) Synthetic product CPSS3:

[0142] (2.1) Esterification: Weigh 0.98 g (0.5 mmol, 1 eq.) of pSar25S, 232 mg (0.6 mmol, 1.2 eq.) of cholesterol, 124 mg (0.6 mmol, 1.2 eq.) of dicyclohexylcarbodiimide (DCC), and 30.5 mg (0.25 mmol, 0.5 eq.) of 4-dimethylaminopyridine (DMAP) and add all of them to a reaction flask. Then, add 10 mL of dichloromethane (DCM), stir at room temperature for 24 h, and filter to obtain the filtrate.

[0143] (2.2) Purification: Add an appropriate amount of diethyl ether to the filtrate and centrifuge for three times. After vacuum drying, 1.08 g of CPSS3 as a white solid was obtained with a yield of 93%.

[0144] Example 3: Performance test of the product CPSS as a drug nanocrystal dispersion and stability.

[0145] In this example, the application potential of CPSS1 with a sarcosine polymerization degree of 14 (ie, n=14) and CPSS2 with a polymerization degree of 28 (ie, n=28) in the preparation of drug nanocrystals was tested.

[0146] In this example, two representative poorly soluble drugs, sirolimus (SIRO) and paclitaxel (PTX), were prepared into nanocrystals. The advantages of TPGS as a stabilizer for drug nanocrystals were evaluated using particle size, PDI, and stability as evaluation indicators. First, paclitaxel nanocrystals were prepared and the dispersion and stability of TPGS, CPSS1, and CPSS2 in the nanocrystals were compared.

[0147] The preparation method is as follows: paclitaxel is dissolved in DMSO (40 mg / mL) to prepare a certain concentration of TPGS / CPSS1 / CPSS2 aqueous solution. The paclitaxel DMSO solution is slowly injected into the TPGS / CPSS1 / CPSS2 aqueous solution using a syringe pump while maintaining high-speed magnetic stirring (1200 rpm). As the paclitaxel DMSO solution is injected, uniform nanoparticles are formed in the aqueous solution. After preparation, the nanoparticles are sonicated for 15 minutes using an ultrasonic cell disruptor (20%, sonication for 2 seconds, and pause for 2 seconds). The sonicated nanocrystal suspension is subjected to ultracentrifugation at 16,000×g for 30 minutes. After the centrifugation, the supernatant is discarded and the nanocrystal precipitate is resuspended in a small amount of water. The nanocrystal suspension is then sonicated for another 5 minutes using an ultrasonic cell disruptor (20%, sonication for 2 seconds, and pause for 2 seconds). The prepared paclitaxel nanosuspension ( Figure 1 ) After dilution with 10 mM sodium chloride, the particle size of the nanocrystals was measured using a particle size potentiometer: the particle size of the nanocrystals prepared with TPGS was 465.54±8.13 nm, with a PDI of 0.121±0.040, the particle size of the nanocrystals prepared with CPSS1 was 146.32±2.17 nm, with a PDI of 0.054±0.039, and the particle size of the nanocrystals prepared with CPSS2 was 168.74±2.55 nm, with a PDI of 0.044±0.009. The results show that the paclitaxel nanocrystals prepared with CPSS1 / CPSS2 have smaller particle sizes and more uniform nanocrystal particles, while the nanocrystals prepared with TPGS have precipitation at the bottom ( Figure 4 ), and a sharp peak appears at the end of the particle size graph ( Figure 5 、 Figure 6 、 Figure 7 ), indicating that CPSS is better than TPGS in dispersing and stabilizing nanocrystals.

[0148] The stability of the prepared paclitaxel nanocrystals was further investigated over time. The paclitaxel nanocrystals prepared by TPGS showed unstable PDI index in a short period of time, while the paclitaxel nanocrystals prepared by CPSS1 / CPSS2 remained stable after storage at 4°C for 50 days, with no significant changes in particle size and PDI ( Figure 12 ). From the above results, it can be seen that different blocks of CPSS not only show a better dispersion effect than TPGS in the preparation of paclitaxel nanocrystals, but also can effectively stabilize the drug nanocrystals and improve the physical stability of their suspensions.

[0149] The same method as above was used to investigate the sirolimus nanocrystals prepared by TPGS / CPSS1 / CPSS2. Figure 2) After diluting to a certain concentration with 10 mM sodium chloride, the particle size and distribution of each nanocrystal suspension were measured using a particle size potentiometer. The results are as follows Figure 4 The stability of the prepared sirolimus nanocrystals was further investigated over time. The sirolimus nanocrystals prepared by TPGS / CPSS1 / CPSS2 remained stable after storage at 4°C for 50 days, with no significant changes in particle size and PDI ( Figure 13 ), it can be seen from the results that the particle size of the nanocrystal suspension prepared by CPSS1 / CPSS2 is smaller than that of the nanocrystal suspension prepared by TPGS.

[0150] Example 4: Determination of the critical micelle concentration (CMC) of the product CPSS.

[0151] In this example, the pyrene fluorescence method was used to determine the critical micelle concentration (CMC) of CPSS. Pyrene is a commonly used fluorescent probe molecule with very low solubility in water. The core of the polymer micelles formed by CPSS is a hydrophobic region, allowing pyrene to solubilize into the CPSS polymer micelles through hydrophobic interactions. When the CPSS concentration is below its critical micelle concentration, CPSS molecules reside on the surface of the solution, i.e., at the gas-liquid interface, and pyrene is not solubilized in the micelles. When the CPSS solubility in the solution reaches its CMC, the gas-liquid interface can no longer accommodate CPSS molecules, and excess CPSS molecules enter the solution to form micelles. Specifically, the lipophilic end of CPSS cholesterol resides within the micelles, while the hydrophilic chains of polysarcosine reside on the surface. As the CPSS concentration in the solution increases further, i.e., above the CMC, more and more CPSS micelles form, solubilizing the poorly soluble pyrene within the micelles. Upon excitation at 335 nm, the fluorescence emission spectrum of pyrene in solution exhibits five electronic vibrational peaks. The intensity ratio of the first and third vibrational peaks, I1 / I3, strongly depends on the polarity of the pyrene environment. Consequently, the polarity of the pyrene environment in solution and in CPSS micelles differs significantly, leading to significant variations in the I1 / I3 ratio. The CMC of CPSS can be calculated by plotting the I1 / I3 ratio against concentration using the tangent method. The critical micelle concentration (CMC) of CPSS 3 (pSar: n=25) was calculated to be 54.65±1.81 μg / mL based on the tangent inflection point.

[0152] The experimental procedure was as follows: 50 μL (10 μg / mL) of a pyrene solution in acetone was added dropwise to a 5.0 mL brown vial and evaporated to dryness in a fume hood. A series of CPSS (pSar: n=25) concentration gradients (0.001, 0.01, 0.1, 1, 5, 10, 50, 100, 250, 500, and 1000 μg / mL) were prepared and added to the pyrene-containing brown vial. The solution was then heated in a 45°C water bath for 1.5 h. After cooling to room temperature, the fluorescence emission spectra of the pyrene-containing and surfactant solutions were scanned using a fluorescence spectrometer, with the surfactant solution serving as a blank control. The temperature was set at 25°C, the excitation wavelength was 335 nm, the excitation slit width was 5 nm, the emission slit width was 2.5 nm, and the emission spectrum was scanned from 365 to 450 nm. The logarithmic value of the concentration of CPSS3 (pSar: n=25) was used as the abscissa, and the ratio of the first fluorescence characteristic peak and the third fluorescence characteristic peak intensity of pyrene was used as the ordinate to draw a curve for fitting. The abscissa value at the intersection of the two fitted straight lines was the CMC, as shown in Figure 2. Figure 11 As shown, in this embodiment, the average value of three times is used as the result value.

[0153] This example verifies that the product CPSS of the present invention has the ability to solubilize drugs.

[0154] Example 5: The product CPSS was used as a stabilizer for drug nanocrystals to measure drug loading.

[0155] In this example, drug loading measurement experiments were performed using drug nanocrystals prepared with CPSS1 (pSar: n=14) / CPSS2 (pSar: n=28).

[0156] In this example, two poorly soluble drugs, sirolimus (SIRO) and paclitaxel (PTX), were selected. The experimental preparation method was the same as that in Example 3. The prepared nanocrystals were then freeze-dried for 24 h and the drug loading was measured using HPLC. For sirolimus nanocrystals prepared using the same experimental parameters and steps, the drug loading rate of TPGS-SIRO nanocrystals was 90.06±10.10%; the drug loading rate of CPSS1-SIRO nanocrystals was 102.36±8.18%; and the drug loading rate of CPSS2-SIRO nanocrystals was 104.8±4.64%. The experimental results show that the ability of CPSS1 / CPSS2 to load sirolimus can be as high as nearly 100%, which is significantly higher than the 90% of TPGS. The use of this type of stabilizer for sirolimus can greatly avoid drug loss. For paclitaxel nanocrystals prepared using the same experimental parameters and steps, the drug loading rate of TPGS-PTX nanocrystals was 65.87±0.99%; the drug loading rate of CPSS1-PTX nanocrystals was 94.34±0.31%; and the drug loading rate of CPSS2-PTX nanocrystals was 96.02±2.00%. The experimental results show that the ability of CPSS1 / CPSS2 to load paclitaxel is similar, both around 95%, which is significantly higher than the 65% of TPGS.

[0157] Example 6, solubilization experiment of the product CPSS on poorly soluble drugs.

[0158] In this example, two poorly soluble drugs were selected, namely, sirolimus (SIRO) and paclitaxel (PTX).

[0159] Excess SIRO and PTX were placed in vials containing CPSS1 (pSar: n=14), CPSS2 (pSar: n=28), or TPGS aqueous solution, and then placed in a constant temperature incubator shaker at 60 rpm for 24 hours to fully dissolve the drug. The samples were then centrifuged at high speed, and the supernatant was collected and the drug content in the supernatant was determined by high-performance liquid chromatography (HPLC). The results are shown in Table 1. Figure 14 As can be seen in the figure, the solubility of SIRO and paclitaxel in water is extremely low, with their concentrations in the supernatant already below the quantification limit of HPLC for both drugs. For SIRO, both TPGS and CPSS can improve its solubility in water to a certain extent, with CPSS2 showing the best effect. However, for PTX, TPGS showed no significant solubilization effect, while CPSS significantly solubilized paclitaxel, demonstrating its superiority over TPGS in solubilizing certain drugs.

[0160] Example 7: The dispersibility and stability of the product CPSS as a component for preparing lipid nanoparticles were tested.

[0161] The polyamino acid polymer material prepared according to the method provided in Example 2 can be used to prepare the complex lipid component of nucleic acid lipid nanoparticles. The product (CPSS) in Example 2 can be used to replace the four different lipid components commonly used in the prior art (ionizable lipids, phospholipids, cholesterol, and polyethylene glycol lipids (PEG lipids)) with three different lipid components (ionizable lipids, phospholipids, CPSS1 (pSar: n=14) / CPSS2 (pSar: n=28)) and four different lipid components (ionizable lipids, phospholipids, cholesterol, and CPSS1 (pSar: n=14) / CPSS2 (pSar: n=28)).

[0162] In this example, the application potential of nucleic acid lipid nanoparticles prepared with CPSS1 (pSar: n=14) / CPSS2 (pSar: n=28) was tested, because the surface charge, particle size, morphology and other factors of lipid nanoparticles are key factors affecting the in vivo process and pharmacokinetic properties of lipid nanoparticle drugs.

[0163] In this example, pCMV-EGFP-3×Linker-MCS-Neo was selected for encapsulation and prepared into nucleic acid lipid nanoparticles. The main experiment was to examine its advantages as lipid nanoparticles using the particle size, PDI and stability of lipid nanoparticles as evaluation indicators.

[0164] The preparation method is:

[0165] 1) Ionizable lipid (SM-102), phospholipid (DSPC), cholesterol (optional), and CPSS1 / CPSS2 were prepared at specific molar ratios into a 10 mg / mL compound lipid in anhydrous ethanol as the solvent. The desired plasmid (or blank lipid nanoparticles) in the citric acid buffer was calculated, with N / P = 8 and a constant flow rate ratio of citric acid buffer / compound phospholipid. Solutions containing the compound lipid in anhydrous ethanol as the organic phase and nucleic acid in citric acid buffer as the aqueous phase were obtained.

[0166] 2) The lipid mixture was injected into the buffer solution using a syringe pump at a flow rate of 50 μL / min by solvent injection, and the mixture was vortexed for 15 s and sonicated on ice for 10 min (20%, 2 s pause, 2 s supersonication);

[0167] 3) The obtained initial product was dialyzed using an 8-14 kDa dialysis bag within 10 minutes, wherein the dialysate was 1× PBS (pH 7.4) at 4°C (the dialysate was at least 50 times the sample) for 2 hours;

[0168] 4) The dialysate was replaced with 1× PBS (pH 7.4) containing 10% sucrose and dialyzed overnight at 4°C; the dialyzed lipid nanoparticles were stored at 4°C.

[0169] As shown in Table 1, when the ratio of organic phase to aqueous phase is 1:1.5 without plasmid encapsulation, the average size and PDI of lipid nanoparticles composed of SM-102 / DSPC / CPSS three components are significantly better than those of lipid nanoparticles composed of SM-102 / DSPC / cholesterol / CPSS four components.

[0170] Table 1

[0171]

[0172] As shown in Table 2, when the ratio of organic phase to aqueous phase is 1:1.5, the average size and PDI of lipid nanoparticles composed of SM-102 / DSPC / CPSS are significantly better than those of lipid nanoparticles composed of SM-102 / DSPC / cholesterol / CPSS, and the encapsulation efficiency (EE%) of the three-component lipid nanoparticles is greater than 95%.

[0173] Table 2

[0174]

[0175] As shown in Table 3, 1 and 2 are lipid nanoparticles without plasmids, and 3 and 4 are lipid nanoparticles with plasmids. When the ratio of organic phase to aqueous phase is 1:3, the lipid nanoparticles composed of SM-102 / DSPC / cholesterol / CPSS also perform well, but the dispersibility is not as good as that of the three-component lipid nanoparticles.

[0176] Table 3

[0177]

[0178] As shown in Table 4, when the ratio of organic phase to aqueous phase is 1:1.5, the lipid nanoparticles composed of SM-102 / DSPC / CPSS have good stability, regardless of the presence or absence of the encapsulated plasmid.

[0179] Table 4

[0180]

[0181] Example 8: A system and method for evaluating polyamino acid polymer compounds

[0182] This embodiment provides a system and method for evaluating the performance of amino acid polymer compounds. According to the present invention, a variety of compounds with different structures can be prepared, and these compounds can be used for aspects such as encapsulation, volume expansion, and emulsification. However, in actual production, it is difficult to conduct experiments on all compounds one by one in a short period of time to verify their effects. In order to enable those skilled in the art to better understand and implement the technical solutions of the present invention, and to better demonstrate the technical effects of the present invention, this embodiment designs an evaluation system that evaluates compounds through data learning and artificial intelligence algorithms, thereby verifying the effects of polymer compounds under the system of the present invention.

[0183] The system includes:

[0184] SMILES processing module: Generates the initial SMILES representation through molecular structure conversion method.

[0185] The specific method is:

[0186] 1. Structure to SMILES: Generate an initial SMILES representation using molecular structure conversion methods. Specifically, based on the input molecular structure, identify atoms and chemical bonds, and perform topological analysis. A molecular graph is constructed, with atoms as nodes and chemical bonds as edges. Connections are recorded, and the SMILES representation is determined through graph traversal.

[0187] 2. Normalization: Use the standardized SMILES generation method (unified molecular representation, fixed atomic order, such as converting benzene: C1=CC=CC=C1 to c1ccccc1) to eliminate redundant symbols.

[0188] 3. Outlier Screening: Remove outliers (e.g., masses <50 or >1000 Da) using molecular weight calculation (calculating total mass based on molecular formula, e.g., ethanol CCO is approximately 46 Da). Examine the distribution of target values ​​using the interquartile range method (based on the upper and lower quartiles of the data, eliminating extreme values, e.g., CMC >1000 μg / mL).

[0189] ChemBERTa encoding module: encodes the processed SMILES representation into ChemBERTa and generates molecular embedding vectors suitable for multi-target regression prediction (CMC, Z-average, PDI).

[0190] The specific method is:

[0191] 1. Tokenization: A dedicated chemical sequence tokenizer is used to segment the SMILES string into chemically defined subunits (tokens). This process breaks the string into units such as atoms (such as C and O), compound atoms, bond symbols, and ring markers. For example, the tokenization of benzene (c1ccccc1) is ['c', '1', 'c', 'c', 'c', 'c', 'c', '1'], with each token representing a fragment of the molecular structure. The tokenizer is based on a pretrained vocabulary to ensure coverage of common SMILES symbols. A [CLS] token is added to the beginning of the sequence for feature aggregation, and a [SEP] token is added to the end of the sequence, resulting in an input such as ['[CLS]', 'c', '1', 'c', 'c', 'c', 'c', 'c', '1', '[SEP]'].

[0192] 2. Input Formatting: Convert the token sequence to a numerical representation. Each token is mapped to a unique integer (token ID) according to the vocabulary. An attention mask is generated, assigning 1 to valid tokens and 0 to padding tokens, allowing the model to focus on valid input. To maintain uniform sequence length, short sequences are padded with [PAD] tokens to a maximum length of 512 tokens. This formatted input is output as a tensor, ready to be fed into the ChemBERTa model for inference.

[0193] Model construction and training module: The backbone network ChemBERTa model structure inherits the Transformer encoder architecture of BERT and is pre-trained on chemical datasets (such as PubChem SMILES) to optimize the representation ability of chemical semantics.

[0194] The specific method is:

[0195] 1. Input the processed molecular embedding vector into ChemBERTa, which includes a pre-trained 12-layer Transformer encoder, to generate context-sensitive embeddings.

[0196] 2. Use the context-dependent embeddings as input to a multi-target regression model based on a multi-layer perceptron for regression prediction, and use the weighted mean square error (MSE) loss to supervise the regression prediction results.

[0197] Specifically, the multi-objective regression model based on the multi-layer perceptron includes:

[0198] Hidden layer 1: For the context-dependent embedding of the input, the input fully connected layer reduces the dimension to 1*512, and then uses the activation function ReLU operation to obtain the hidden layer feature one;

[0199] Hidden layer 2: Hidden layer feature 1 is input into the fully connected layer to reduce the dimension to 1*256, and then the activation function ReLU is used to obtain hidden layer feature 2;

[0200] Hidden layer 3: Hidden layer feature 2 is input into the fully connected layer to reduce the dimension to 1*128, and then the activation function ReLU is used to obtain hidden layer feature 3;

[0201] Regression head - CMC: The three-input regression head of the hidden layer features is passed through the fully connected layer to obtain the CMC prediction result with a dimension of 1*1;

[0202] Regression head - Z-average: The hidden layer features are input into the three-input regression head, and the fully connected layer is used to obtain the Z-average prediction result with a dimension of 1*1;

[0203] Regression head-PDI: The hidden layer features are input into the three-input regression head, and the PDI prediction result with a dimension of 1*1 is obtained through the fully connected layer.

[0204] Specifically, the weighted mean square error (Weighted MSE) loss, for multi-target regression tasks, assuming there are K target variables (such as K = 3, corresponding to CMC, Z-average, PDI), the calculation formula of weighted MSE is:

[0205]

[0206] in Indicates all of the goals goals, For the corresponding weights, in this embodiment, the weights are all taken to be 1. for The mean square error between the target result and the true value is calculated as:

[0207]

[0208] in For all sample sizes, and represent the predicted value and the true value respectively.

[0209] After the model parameters are fixed through additional pre-training data training, the processed unknown molecule embedding vector is processed according to SMILES - ChemBERTa encoding - model prediction to obtain the predicted results of CMC, Z average, and PDI.

[0210] Table 5 lists the different indicators and evaluation results of some compounds.

[0211]

[0212] Note: Chemical formula I: ;

[0213] Chemical formula II: .

[0214] As can be seen from the above table, the evaluation system of the present invention can be used to evaluate core indicators such as CMC, Z-average nanoparticle size, and PDI value for polymer compounds obtained by selecting different groups. This provides a preliminary theoretical basis for subsequent experiments, reduces experimental costs, and can more accurately identify the appropriate preparation direction.

[0215] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A polyamino acid polymer material, characterized in that: Its chemical formula is shown in Formula I: Formula I, Here, n is 14, 25, or 28.

2. A polyamino acid polymer material according to claim 1, characterized in that: The auxiliary material of the polyamino acid polymer material includes at least one selected from the group consisting of organic solvents DMSO, DMF, DMAc, DCM, ethyl ether, methyl tert-ether, isopropyl ether, n-heptane, n-hexane and petroleum ether.

3. A polyamino acid polymer material according to claim 1, characterized in that: The synthetic route of the polyamino acid polymer material is as follows: Here, n is 14, 25, or 28.

4. Use of the polyamino acid polymer material according to any one of claims 1 to 3 in dissolution.

5. Use of the polyamino acid polymer material according to any one of claims 1 to 3 in preparing a drug delivery carrier.

6. Use of the polyamino acid polymer material according to any one of claims 1 to 3 in emulsification.

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