Ionizable lipid compounds targeting atherosclerotic cells, delivery vehicle compositions, pharmaceutical compositions, drug-loaded complexes, pharmaceutical formulations, uses and methods of manufacture

Abstract

The present application relates to ionizable lipid compounds targeting atherosclerotic cells, delivery carrier compositions, pharmaceutical compositions, drug-loaded complexes, pharmaceutical preparations, applications and preparation methods. The ionizable lipid compounds targeting atherosclerotic cells successively comprise two unsaturated fatty acyl groups with anti-inflammatory activity, an N atom branching center, a divalent linking group G containing an ionizable N atom, and a structural unit R capable of targeting atherosclerotic cell functions 03 . The ionizable lipid compounds can target atherosclerotic cells and have anti-inflammatory activity, which can not only improve plaque inflammation as anti-inflammatory drugs, but also serve as structural components of drug complexes such as lipid nanoparticles, and can co-deliver bioactive ingredients such as small molecules, polypeptides, and gene drugs, and can be used for synergistic treatment of atherosclerosis.

Description

Technical Field

[0001] This application relates to the field of biomedical technology, and further to ionizable lipid compounds targeting atherosclerotic cells, their applications and preparation methods, and even further to ionizable lipid compounds targeting atherosclerotic cells, delivery carrier compositions, pharmaceutical compositions, drug-loaded complexes, pharmaceutical formulations, their applications and preparation methods. Background Technology

[0002] In recent years, cardiovascular disease has surpassed infectious diseases as the leading cause of death and disability worldwide. Atherosclerosis, accompanied by plaque accumulation, leads to restenosis of blood vessels, reducing blood flow and is widely recognized as a pressing issue in cardiovascular disease. Currently, clinical treatment for atherosclerosis primarily relies on conventional lipid-lowering, anticoagulant, and thrombolytic drugs. For example, statins limit cholesterol synthesis in the liver, lowering systemic cholesterol levels, while fibrates enhance lipoprotein lipase activity, inhibiting triglyceride synthesis in the liver. However, these drugs suffer from poor targeting and significant off-target effects, resulting in unsatisfactory clinical outcomes. Currently, there is a lack of effective intervention strategies for targeted therapy against plaque sites. Therefore, exploring treatment strategies targeting atherosclerotic plaques is urgently needed.

[0003] Drug delivery systems (DDS) are crucial tools for achieving targeted and precision medicine. With the official approval of Comirity (BNT162b2) and Spikevax (mRNA-1273) mRNA COVID-19 vaccines, lipid nanoparticles (LNPs) have emerged as the safest and most efficient drug delivery system, applicable to the delivery of various small / large molecules, peptides, and gene drugs. However, there are currently no suitable LNP formulations for anti-atherosclerotic therapy. Ionizable lipids are an important component of LNPs, effectively addressing the safety issues of traditional cationic lipids such as DOTAP, including cytotoxicity and pro-inflammatory reactions, while also achieving highly efficient nucleic acid transfection. Ionizable lipids, represented by SM-102 and ALC-0135, remain neutral under physiological conditions but protonate in acidic environments, leading to endosomal rupture and drug release. However, the number of approved ionizable lipid materials is limited, their structures are simple, and most lack biological activity, often requiring the addition of active drugs to achieve therapeutic goals, making it difficult to meet complex and varied clinical needs. For atherosclerosis, single-drug delivery is often unsatisfactory, while encapsulating multi-component drugs increases development difficulty and clinical application risks. Current modifications to ionizable lipid materials mainly focus on target modification, neglecting the development of their intrinsic functions, which significantly limits their development in the biomedical field.

[0004] Therefore, it is necessary to develop new delivery formulations that target atherosclerosis. Summary of the Invention

[0005] According to various embodiments and examples of this application, this application provides an ionizable lipid compound targeting atherosclerotic cells, a delivery carrier composition, a pharmaceutical composition, a drug-loaded complex, a pharmaceutical formulation, its application, and a method of preparation. This ionizable lipid compound can target atherosclerotic cells and possesses anti-inflammatory activity. It can be used as an anti-inflammatory drug to improve plaque inflammation, and can also serve as a structural component of complexes such as lipid nanoparticles. It can co-deliver bioactive components such as small molecules, peptides, and gene drugs, and can be used for synergistic treatment of atherosclerosis, but is not limited to this application.

[0006] In some embodiments of the first aspect of this application, an ionizable lipid compound that targets atherosclerotic cells is provided, wherein the ionizable lipid compound is a compound with the structure shown in formula (I);

[0007]

[0008] In formula (I),

[0009] R1- and R2- each independently form an unsaturated fatty acyl group with anti-inflammatory activity with the carbonyl group;

[0010] Z1 and Z2 are each independently O, NH or S;

[0011] L1 and L2 are each independently an alkylene group with 1 to 3 carbon atoms in the main chain;

[0012] G is a divalent aliphatic hydrocarbon group containing one or more ionizable N atoms;

[0013] R3 includes structural units R that can target the function of atherosclerotic cells. 03 .

[0014] In some other embodiments of the first aspect of this application, an ionizable lipid compound that targets atherosclerotic cells is provided, said ionizable lipid compound having the structure shown in formula (I);

[0015] In formula (I),

[0016] R1- and R2- independently form an unsaturated fatty acyl group with the carbonyl group, derived from α-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, medelic acid, or sinapic acid.

[0017] Z1 and Z2 are each independently O, NH or S;

[0018] L1 and L2 are each independently C 1-3 Alkylene;

[0019] G is a divalent aliphatic hydrocarbon group containing one or more ionizable N atoms;

[0020] R3 includes structural units R that have the function of targeting atherosclerotic cells. 03 .

[0021] In some embodiments of the second aspect of this application, a delivery carrier composition is provided, by molar percentage, comprising 20% ​​to 70% of an ionizable lipid compound, 0% to 60% of a steroid, 0% to 30% of an auxiliary lipid, 0% to 10% of a polyethylene glycol-modified lipid, and 0% to 15% of other pharmaceutical excipients; or, by mass percentage, the delivery carrier composition comprises 20% to 90% of an ionizable lipid compound, 0% to 50% of a steroid, 0% to 50% of an auxiliary lipid, 0% to 20% of a polyethylene glycol-modified lipid, and 0% to 80% of other pharmaceutical excipients.

[0022] Wherein, the ionizable lipid compound is at least one of the ionizable lipid compounds targeting atherosclerotic cells described in the first aspect of this application;

[0023] Optionally, the delivery carrier composition comprises at least one of the steroid, the auxiliary lipid, and the polyethylene glycol-modified lipid;

[0024] Optionally, the auxiliary lipids include one or more of phosphatidylethanolamines (non-limiting examples such as diacylphosphatidylethanolamine), phosphatidylcholine derivatives (non-limiting examples such as diacylphosphatidylcholine), betaine lipids (DGTS), and sphingomyelins (SM).

[0025] In some embodiments of another aspect of this application, a drug delivery carrier is provided, comprising the delivery carrier composition described in the second aspect of this application.

[0026] In some embodiments, the drug delivery carrier is a liposome, lipid nanoparticle, polymer nanoparticle, nanomicelle, or nanoemulsion drug carrier.

[0027] In some embodiments of the third aspect of this application, a bioactive composition, a bioactive complex, or a bioactive formulation is provided, the bioactive formulation comprising the bioactive complex; the bioactive composition or the bioactive complex independently comprises a bioactive ingredient and an ionizable lipid compound targeting atherosclerotic cells as described in the first aspect of this application;

[0028] Optionally, the bioactive ingredient includes a pharmaceutical ingredient, the bioactive composition is a pharmaceutical composition, the bioactive complex is a drug-loaded complex, and the bioactive formulation is a pharmaceutical formulation.

[0029] In other embodiments of the third aspect of this application, a pharmaceutical composition, a drug-loaded complex, or a pharmaceutical formulation is provided, the pharmaceutical formulation comprising a drug-loaded complex; the pharmaceutical composition or the drug-loaded complex independently comprises a bioactive ingredient and an ionizable lipid compound targeting atherosclerotic cells as described in the first aspect of this application.

[0030] In some embodiments of the fourth aspect of this application, the ionizable lipid compound targeting atherosclerotic cells described in the first aspect of this application is provided for use in the preparation of a drug delivery carrier or drug-loaded complex, or in the preparation of a medicament for treating a disease; or, the ionizable lipid compound targeting atherosclerotic cells described in the first aspect of this application, the delivery carrier composition described in the second aspect of this application, the drug delivery carrier (such as liposomes or lipid nanoparticles) described in one aspect of this application, or the drug composition or drug-loaded complex described in the third aspect of this application is used in the preparation of a pharmaceutical formulation for treating a disease or in the preparation of a medicament for treating a disease.

[0031] The diseases to be treated include at least one of the following: treatment of chronic inflammatory diseases, targeted therapy for inflammatory diseases, targeted therapy for atherosclerosis, treatment of atherosclerotic inflammation, treatment of familial hyperlipidemia, treatment of atherosclerotic lipid deposition, treatment of hypertension caused by atherosclerosis, treatment of atherosclerotic stroke, and treatment of atherosclerotic vascular damage; the inflammatory diseases include plaque inflammation; and the chronic inflammatory diseases include cardiovascular diseases.

[0032] Optionally, the disease includes at least one of immune-related inflammatory cardiovascular disease, atherosclerotic inflammation, atherosclerotic myocardial infarction, and atherosclerotic stroke.

[0033] In some embodiments of another aspect of this application, the use of the ionizable lipid compounds targeting atherosclerotic cells described in the first aspect of this application is provided in the preparation of liposomes, lipid nanoparticles, polymer nanoparticles, nanomicelles, or nanoemulsion drug carriers.

[0034] In some embodiments of another aspect of this application, the use of the ionizable lipid compounds targeting atherosclerotic cells described in the first aspect of this application is provided in the preparation of drug-loaded liposomes, drug-loaded lipid nanoparticles, drug-loaded polymer nanoparticles, drug-loaded nanomicelles, or nanoemulsion pharmaceutical formulations.

[0035] In some embodiments of another aspect of this application, a method for treating inflammation in vitro is provided, comprising administering to cells an ionizable lipid compound as described in the first aspect of this application, a delivery carrier composition as described in the second aspect of this application, a drug delivery carrier (such as liposomes or lipid nanoparticles) as described in one aspect of this application, a drug composition, a drug-loaded complex, or a drug formulation.

[0036] In some embodiments of another aspect of this application, a method of treating a disease is provided, comprising administering to a subject a therapeutically effective amount of the ionizable lipid compound described in the first aspect of this application, the delivery carrier composition described in the second aspect of this application, the drug delivery carrier (such as liposomes or lipid nanoparticles) described in one aspect of this application, the drug composition described in the third aspect of this application, a drug-loaded complex, or a drug formulation. The definition of a disease can be found in the fourth aspect of this application.

[0037] In some embodiments of another aspect of this application, methods for preparing the aforementioned ionizable lipid compounds, delivery carrier compositions, drug delivery carriers, drug compositions, drug complexes, or drug formulations are provided.

[0038] In some embodiments of the fifth aspect of this application, a method for preparing a drug-loaded complex or pharmaceutical preparation is provided, comprising the following steps:

[0039] The components of the delivery carrier composition described in the second aspect of this application are dissolved in ethanol to prepare a lipid ethanol solution;

[0040] The lipid ethanol solution is mixed with a drug aqueous solution using any one of microfluidic methods, thin-film hydration combined with electroporation, and pressure homogenization combined with shearing to prepare a drug-loaded complex or a liquid pharmaceutical formulation including a drug-loaded complex; the pharmaceutical formulation is as defined in the third aspect of this application, and the drug-loaded complex is the drug-loaded complex in the pharmaceutical formulation described in the third aspect of this application; wherein the drug aqueous solution is an aqueous solution containing bioactive components.

[0041] Optionally, the bioactive component is a nucleic acid-like substance;

[0042] Optionally, the drug-loaded complex is a lipid nanoparticle, a liposome, or a lipid nanoemulsion;

[0043] Optionally, the method for preparing the pharmaceutical formulation further includes the step of drying the liquid pharmaceutical formulation to form a solid pharmaceutical formulation.

[0044] In some embodiments, the liquid formulation contains a dispersion liquid; the dispersion liquid includes water and may or may not include a buffer; wherein the buffer includes one or more of a salt component of a phosphate buffer solution, trimethylolpropanesulfonic acid, sodium citrate, citric acid, sodium dihydrogen phosphate, potassium dihydrogen phosphate, and 4-hydroxyethylpiperazine ethanesulfonic acid.

[0045] This ionizable lipid compound can target atherosclerotic cells and has anti-inflammatory activity. It can be used as an anti-inflammatory drug to improve plaque inflammation, and can also serve as a structural component of bioactive complexes (such as drug complexes) such as lipid nanoparticles. It can co-deliver bioactive components such as small molecules, peptides, and gene drugs, and can be used for synergistic treatment of atherosclerosis, but is not limited to this application.

[0046] This ionizable lipid compound can promote the targeted accumulation of the encapsulated bioactive components in atherosclerotic plaques.

[0047] The aforementioned ionizable lipid compounds targeting atherosclerotic cells can be applied in the treatment of atherosclerosis, and are of significant importance in promoting the cure of cardiovascular and other chronic inflammatory diseases. On the one hand, compared with traditional ionizable lipid molecules, these ionizable lipid compounds have anti-inflammatory effects, which can improve the inflammatory response at the lesion site and target and accumulate at the lesion site to achieve the purpose of precise treatment. On the other hand, compared with co-delivering anti-inflammatory drugs, these ionizable lipid compounds can be used as components of drug delivery systems such as liposomes, lipid nanoparticles, and polymer micelles, directly participating in the structural formation of drug-loaded complexes. This can enhance the effects of nucleic acid transfection and drug delivery, while also mitigating the unknown risks in drug development to a certain extent.

[0048] This ionizable lipid compound can be used for the treatment of atherosclerotic inflammation.

[0049] This ionizable lipid compound can be applied to the targeted therapy of inflammatory diseases. It comprises monounsaturated or polyunsaturated fatty acid tail chains with anti-inflammatory activity, which can inhibit the secretion of inflammatory factors by cells, thereby reducing the inflammatory response at the disease site. The ionizable lipid compound also has a head group targeting atherosclerosis, which can increase the targeted accumulation of lipids at the disease site, improve drug delivery via drug delivery carriers or drug-loaded complexes, enhance the accumulation of the drug-loaded complex itself at the lesion site, and strengthen both the efficacy of the drug and the anti-inflammatory effect of the drug-loaded complex.

[0050] This ionizable lipid compound can be used as a component of a pharmaceutical composition to prepare a drug formulation. Its ionization property allows for the delivery of bioactive therapeutic drugs, such as nucleic acids, thereby improving the safety and transfection efficiency of nucleic acid drugs in vivo. Exemplary drug-loaded complexes or drug formulations may include lipid nanoparticles, liposomes, and lipid nanovesicles. Pharmaceutical compositions, drug-loaded complexes, and drug formulations incorporating this ionizable lipid compound offer advantages such as inhibiting inflammatory responses, promoting targeted drug aggregation at lesions, and achieving high nucleic acid transfection efficiency.

[0051] Loading bioactive components onto the aforementioned ionizable lipid compounds with anti-inflammatory activity and drug delivery carriers (such as liposomes, lipid nanoparticles, and other complexes) can yield drug-loaded complexes or drug formulations that can deliver bioactive components into the body. This allows the loaded or delivered bioactive components to exert their effects more effectively and can play an advantage in targeted therapy for immune-related inflammatory cardiovascular diseases.

[0052] Details of one or more embodiments or examples of this application are set forth in the following drawings and description. Other features, objects, and advantages of this application will become apparent from the specification, drawings, and claims. Attached Figure Description

[0053] To more clearly illustrate the technical solutions in the embodiments and examples of this application, and to more completely understand this application and its beneficial effects, the accompanying drawings used in the description of the embodiments or examples will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of this application. Those skilled in the art can obtain other drawings based on these drawings without any creative effort.

[0054] In the attached diagram:

[0055] Figure 1 The results show the particle size of drug nanocarrier complexes prepared with different formulations in some embodiments of this application.

[0056] Figure 2 The graph shows the potential test results of drug nanocarrier complexes prepared with different formulations in some embodiments of this application. The vertical axis represents the zeta potential (unit: mV).

[0057] Figure 3 The pKa values ​​of drug nanocarrier complexes prepared with different formulations in some embodiments of this application are shown. The horizontal axis represents pH and the vertical axis represents relative fluorescence intensity (RFI).

[0058] Figure 4 The following are test results of drug nanocarrier complexes prepared with different formulations in some embodiments of this application being targeted to atherosclerotic cells and thus taken up: (A) is the result of flow cytometry quantitative test of cell uptake, with the mean fluorescence intensity (MFI) on the ordinate; (B) is the result of fluorescence confocal test of cell uptake, "nucleous & DiI" indicates that nuclear staining and DiI staining were performed simultaneously.

[0059] Figure 5 These are test results of the gene silencing effect of drug nanocarrier complexes prepared with different formulations in some embodiments of this application.

[0060] Figure 6 These are test results of in vivo IL-6 and TNF-α inflammatory factor secretion levels of drug nanocarrier complexes prepared with different formulations in some embodiments of this application.

[0061] Figure 7 These are test results of in vivo inhibition of atherosclerotic plaque development by drug nanocarrier complexes prepared with different formulations in some embodiments of this application.

[0062] Figure 8 For some embodiments of this application, compounds f and 1 1 1H NMR spectrum, with the horizontal axis representing chemical shift (f1, in ppm).

[0063] Figure 9 The mass spectrometry results of compounds f and 1 in one embodiment of this application are shown. The horizontal axis represents the mass-to-charge ratio (m / z), and the vertical axis represents the ionic intensity. Detailed Implementation

[0064] The present application will be further described in detail below with reference to the accompanying drawings, embodiments, and examples. It should be understood that these embodiments and examples are for illustrative purposes only and are not intended to limit the scope of the present application. The purpose of providing these embodiments and examples is to enable a more thorough and comprehensive understanding of the disclosure of the present application. It should also be understood that the present application can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various modifications or alterations without departing from the spirit of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. For example, features described or illustrated as part of one embodiment can be combined in a suitable manner in another embodiment to produce new embodiments. Furthermore, numerous details are set forth in the following description to provide a fuller understanding of the present application. It should be understood that the present application can be implemented without one or more of these details.

[0065] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the specification of this application is for descriptive purposes only and is not intended to be limiting of the application.

[0066] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0067] The terms “and / or,” “or / and,” and “and / or” as used herein include any one of two or more of the related listed items, as well as any and all combinations of the related listed items. “Any and all combinations” includes any two related listed items, any more related listed items, or a combination of all related listed items. For example, “A and / or B” includes three parallel options: A, B, and “a combination of A and B.”

[0068] In this application, the terms "multiple", "various", "multiple times", "several", "several", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more or more.

[0069] In this application, unless otherwise specified, "one or more" means any one of the listed items or any combination of the listed items. Similarly, "one or more" and other instances that otherwise indicate "one or more" shall be understood in the same way unless otherwise specified.

[0070] In this application, the word "suitable" in "suitable combination", "suitable method", "any suitable method" etc., shall be defined as being able to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0071] In this application, terms such as "preferred," "better," "more suitable," "more preferable," "better," and "more advantageous" are merely descriptions of more effective implementation methods or embodiments, and should be understood not to limit the scope of protection of this application. If multiple "preferred" terms appear in a technical solution, unless otherwise specified and there are no contradictions or mutual constraints, each "preferred" term shall be independent.

[0072] In this application, terms such as "further," "even more," "particularly," "for example," "like," "example," and "exemplary" are used for descriptive purposes to indicate that different technical solutions preceding and following each other are related in terms of their coverage, but should not be construed as limiting the preceding technical solution or restricting the scope of protection of this application. In this application, unless otherwise specified, A (e.g., B) indicates that B is a non-limiting example of A, and it can be understood that A is not limited to B.

[0073] In this application, "optionally," "optionally," and "optional" mean that something is optional, that is, it is selected from either "present" or "absent." If multiple "options" appear in a technical solution, unless otherwise specified and there are no contradictions or mutual constraints, each "option" is independent. Unless otherwise specified, the descriptions such as "optionally include" and "optionally contain" in this application, taking "optionally include" as an example, mean "may include or not include." "Optional component X" indicates whether component X exists or does not exist.

[0074] The terms “containing,” “comprising,” and “including” as used in this application are synonyms and are inclusive or open-ended, not excluding additional, uncited members or features. Members or features include, for example, materials or components, structures, elements, instruments, etc.; non-limiting examples of members or features include actions, conditions under which actions occur, timing, states, etc.

[0075] In this application, the technical features or solutions described in open-ended language include both closed-ended technical features or solutions consisting of the listed contents and open-ended technical features or solutions that include the listed contents.

[0076] In this application, the terms "room temperature" and "normal temperature" generally refer to 4℃ to 35℃, for example, 20℃ ± 5℃. In some embodiments of this application, "room temperature" or "normal temperature" refers to 20℃ to 30℃.

[0077] In this application, if the unit for a data range is only followed by the right endpoint, it indicates that the units for the left and right endpoints are the same. For example, 3~5h means that the units for the left endpoint "3" and the right endpoint "5" are both h (hours), and both have the same meaning as 3h~5h. Furthermore, similar descriptions involving other parameters such as temperature and size also apply to the above understanding.

[0078] All references to documents mentioned in this application are incorporated herein by reference as if each document were individually incorporated by reference. Unless they conflict with the inventive purpose and / or technical solution of this application, all cited documents are incorporated herein by reference in their entirety and for all purposes. When citing documents in this application, the definitions of relevant technical features, terms, nouns, phrases, etc., are also incorporated herein by reference. When citing documents in this application, examples and preferred embodiments of the cited technical features may also be incorporated herein by reference, but only to the extent that they enable the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or modifications shall be made adaptably to the description in this application.

[0079] In this application, where the method flow involves multiple steps, unless otherwise explicitly stated herein, there is no strict order restriction on the execution of these steps; they can be executed in any order other than those described. Moreover, any step may include multiple sub-steps or multiple stages, which are not necessarily completed at the same time, but can be executed at different times, and their execution order is not necessarily sequential, but can be performed alternately or simultaneously with other steps or parts of the sub-steps or stages of other steps.

[0080] In this application, the exemplary descriptions such as "in some implementations (or embodiments)" and "in one implementation (or embodiment)" may cover, but are not limited to, the following meanings: these solutions can be combined with other solutions in a suitable manner to form new technical solutions.

[0081] In this application, the terms "first aspect," "second aspect," "third aspect," "fourth aspect," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first," "second," "third," "fourth," etc., serve only as a non-exhaustive enumeration and should be understood not to constitute a closed limitation on quantity.

[0082] In this application, when numerical intervals (i.e., numerical ranges) are involved, unless otherwise specified, the distribution of selectable numerical values ​​within the numerical interval is considered continuous, and includes the two endpoints of the numerical interval (i.e., the minimum and maximum values), as well as every numerical value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that numerical interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints, which is equivalent to directly listing every integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be merged. In other words, unless otherwise specified, the numerical ranges disclosed herein should be understood to include any and all subranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, percentage, ratio, etc. The term "numerical interval" can be broadly included to include numerical interval types such as percentage intervals, ratio intervals, and proportion intervals.

[0083] In the following examples, the measurement parameters of the raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0084] In this application, unless otherwise specified, any numerical values ​​or ranges refer to reasonable approximations of their values, and ranges of values ​​encompass reasonable approximations of their two endpoints. Those skilled in the art will understand that fluctuations resulting from these approximations are all within the range indicated by the numerical range. That is, in this application, unless otherwise specified, "N1" and "approximately N1" have the same meaning and can be used interchangeably, as do "N1~N2" and "approximately N1 to approximately N2," where N1 and N2 are two unequal values. In some cases, due to reasonable deviations allowed in the art, instrument control precision, or one or more other factors, approximate values ​​within the approximation range should also be included within the range indicated by the numerical range. For example, "temperature is 20℃~30℃" can be understood as "approximately 20℃~approximately 30℃"; further, taking "20℃" as the endpoint and its approximation as ±1℃, approximate values ​​such as 19℃ and 19.5℃ within the approximation range indicated by "approximately 20℃" should also be included in the range indicated by 20℃~30℃. For example, "mass percentage is 30%~70%" can be understood as "approximately 30% to approximately 70%"; further, taking "70%" as the left endpoint and "70%" as the right endpoint and both approximations as ±0.1%, approximate values ​​such as 29.9% and 70.1% within the approximation range indicated by "approximately 30% to approximately 70%" should also be included in the range indicated by 30%~70%.

[0085] In this application, unless otherwise specified, "about" means within a certain range above and below the stated number, and the fluctuation range may vary depending on the type and value of the stated number. For example, a range of ±10%, ±5%, ±2%, ±1%, etc., is permissible. For example, taking "about 20℃" with an approximation of ±0.5℃ as an example, approximate values ​​such as 19℃ and 19.5℃ within the approximation range indicated by "about 20℃" should also be included in the range indicated by "about 20℃". For example, "about 600Da" can be selected from 600±60Da, 600±30Da, etc.

[0086] In this application, the terms "molecular weight," "average molecular weight," "relative atomic mass," or "relative molecular mass" for compounds or polymers, unless otherwise specified, refer to the relative mass measured in Daltons (Da), where 1 Dalton equals... 12 One-twelfth of the atomic mass of carbon. Unless otherwise specified, the average molecular weight of polymers is greater than or equal to (≥) 1000 Da, and the molecular weight of small molecule compounds is less than 1000 Da. 1 kDa = 1000 Da.

[0087] In this application, unless otherwise specified, the number of carbon atoms in a compound or group may be described using the subscript "C". For example, "C 15-21 "C" indicates that it has 15 to 21 carbon atoms.15-21 "Each time it appears, it can be independently classified as C." 15 C 16 C 17 C 18 C 19 C 20 or C21 For example, “C” 1-6 "" indicates that it has 1 to 6 carbon atoms, "C 1-6 "Each time it appears, it can be independently classified as C1, C2, C3, C4, C5, or C6. For example, 'C'..." 1-3 "" indicates that it has 1 to 3 carbon atoms, "C 1-3 Each time it appears, it can be independently classified as C1, C2, or C3. For example, "C..." 1-6 "alkyl" indicates an alkyl group having 1 to 6 carbon atoms, "C 1-6 Each time the word "alkyl" appears, it can independently be a C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, or C6 alkyl. For example, "halogenated C..." 1-6 "alkyl" refers to a halogenated product formed by replacing an alkyl group having 1 to 6 carbon atoms with one or more halogens. 1-6 Each time the word "alkyl" appears, it can independently be a halo-C1 alkyl, halo-C2 alkyl, halo-C3 alkyl, halo-C4 alkyl, halo-C5 alkyl, or halo-C6 alkyl. For example, "amino-substituted C..." 1-6 "Alkyl" indicates an alkyl group having 1 to 6 carbon atoms that is substituted with one or more amino groups. "Amino-substituted C" refers to the form where an alkyl group has 1 to 6 carbon atoms and is replaced by one or more amino groups. 1-6 Each time the word "alkyl" appears, it can be an amino-substituted C1 alkyl, amino-substituted C2 alkyl, amino-substituted C3 alkyl, amino-substituted C4 alkyl, amino-substituted C5 alkyl, or amino-substituted C6 alkyl, which can be independent of each other.

[0088] Unless otherwise stated in this application, the following shall apply: This represents a covalent single bond connection site.

[0089] In this application, the term "hydrocarbon group" refers to a group composed of carbon and hydrogen atoms, specifically a group formed by the loss of at least one hydrogen atom from a corresponding hydrocarbon, with a corresponding covalent bonding site formed at the position where the hydrogen atom was lost. Unless the valence state of the "hydrocarbon group" is directly or indirectly specified, it generally refers to a monovalent hydrocarbon group. For example, when "alkyl" is mentioned, unless the valence state is directly or indirectly specified, it generally refers to a monovalent alkyl group. A group formed by the loss of two hydrogen atoms from a corresponding hydrocarbon is a divalent hydrocarbon group.

[0090] In this application, the term "alkyl" refers to a chain hydrocarbon group with a saturated structure consisting of carbon and hydrogen atoms. Alkyl groups do not contain carbon-carbon double bonds and do not include cyclic structures.

[0091] In this application, the term "alkylene" refers to a hydrocarbon group with two monovalent groups, derived from an alkyl group by losing one hydrogen atom; therefore, alkylene is a divalent group and has a saturated structure, not containing a carbon-carbon double bond. For example, "C 1-3 "Alkylene" refers to an alkyl group containing 1 to 3 carbon atoms, and each time it appears, it can be independently C1 alkylene, C2 alkylene, or C3 alkylene.

[0092] In this application, the term "aliphatic hydrocarbon group" refers to a hydrocarbon group that does not have aromatic properties, that is, aliphatic hydrocarbon groups do not include aromatic cyclic structures.

[0093] In this application, the term "carbon-carbon double bond" refers to a structure having a carbon skeleton of "-C=C-", and a non-limiting example of a carbon-carbon double bond is -CH=CH- or -CH=CH2.

[0094] In this application, "carbonyl" is -C(=O)-, "carboxyl" is -COOH, and "hydroxyl" is -OH.

[0095] In this application, the terms "straight-chain structure" and "branched-chain structure" both refer to ring-free structures. In a straight-chain structure, the main chain atoms are connected in series, while in a branched-chain structure, there are branching points.

[0096] In this application, unless otherwise specified, "small molecule organic compound" generally refers to an organic compound with a molecular weight of less than or equal to 1000 Da.

[0097] In this application, unless otherwise specified, "peptide" and "protein" are both polymers formed by multiple amino acids linked sequentially by peptide bonds. A polypeptide contains 2 to 50 amino acid units, and a protein contains more than 50 amino acid units. In this application, unless otherwise specified, "peptide bond" refers to the connection between the N-terminus and C-terminus of an amino acid in the manner of "-NH-C(=O)-" or "-NC(=O)-", or the connection between the C-terminus and N-terminus in the manner of "-C(=O)-NH-" or "-C(=O)-N-".

[0098] Although atherosclerosis was initially thought to be caused by lipid buildup in the arterial wall, increasing evidence shows that inflammatory responses mediated by innate and adaptive immunity under pathological conditions persist throughout the entire process of vascular lesions, including foam cell formation, fibrous plaque formation and rupture, becoming a direct cause of atherosclerosis. Multiple important inflammatory signaling pathways have been reported to be involved in atherosclerotic lesions, such as TLR, NLRP3 inflammasome, PCSK9, JAK / STAT, MAPK, NF-κBp65, and CD40 / CD40L. Dozens of inflammatory markers are involved in atherosclerosis, including vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), E-selectin, and monocyte chemoattractant protein-1 (MCP-1). Furthermore, pro-inflammatory cytokines such as IL-1β, IL-6, IL-17, AIM2, and TNF-α can further exacerbate the inflammatory response at atherosclerotic plaque sites and induce immune cell aggregation. However, in contrast to the aforementioned pro-inflammatory cytokines, IL-37 and IL-1Ra are key anti-inflammatory agents in the arterial wall. They compete with IL-1β for binding to IL-1 receptors, inhibit receptor activation, and thus reduce the inflammatory response.

[0099] Given the importance of inflammation in atherosclerosis, large-scale clinical trials have tested anti-inflammatory drugs. The positive results of cannabinoids in the CANTOS trial marked a milestone in proving the "inflammatory hypothesis of atherosclerosis." Similarly, the COTCOL trial showed that low-dose colchicine, an anti-inflammatory drug, has a CRP-independent effect and can reduce cardiovascular risk events. However, while these anti-inflammatory drugs alleviate inflammation levels in atherosclerosis patients to some extent, they suffer from poor target specificity and severe off-target effects, posing new challenges to atherosclerosis treatment based on the anti-inflammatory theory. The complexity of plaque inflammation suggests that a "one-size-fits-all" approach to anti-inflammatory treatment may be ineffective at different stages of atherosclerosis. Therefore, developing precise interventions that target plaque inflammation is a key strategy for achieving anti-inflammatory therapy for atherosclerosis.

[0100] According to various embodiments and examples of this application, this application relates to ionizable lipid compounds targeting atherosclerotic cells, their applications, and preparation methods. It further provides an ionizable lipid compound targeting atherosclerotic cells, a delivery carrier composition, a bioactive composition, a bioactive complex, a bioactive formulation, its application, and preparation methods. More specifically, it provides an ionizable lipid compound targeting atherosclerotic cells, a delivery carrier composition, a pharmaceutical composition, a drug-loaded complex, a pharmaceutical formulation, its application, and preparation methods. This ionizable lipid compound can target atherosclerotic cells and possesses anti-inflammatory activity. It can be used as an anti-inflammatory drug to improve plaque inflammation, and can also serve as a structural component of complexes such as lipid nanoparticles. It can co-deliver bioactive components such as small molecules, peptides, and gene drugs, and can be used for synergistic treatment of atherosclerosis, but is not limited to this application.

[0101] In some embodiments, the ionizable lipid compound targeting atherosclerotic cells sequentially comprises two unsaturated fatty acyl groups with anti-inflammatory activity, an N-atom branching center, a divalent linker G containing an ionizable N-atom, and a structural unit R capable of targeting the function of atherosclerotic cells. 03 .

[0102] In this application, unless otherwise specified, "delivery carrier composition" refers to a composition that can be used as a delivery carrier or can be used to form a delivery carrier. Unless otherwise specified, "composition" as used herein includes a variety of components that can together form a single mixed system or can be separated into multiple packages in any suitable manner but can be mixed to form a delivery carrier.

[0103] In this application, unless otherwise specified, "bioactive composition" refers to a composition containing a bioactive ingredient. A bioactive composition may be a pharmaceutical composition.

[0104] In this application, unless otherwise specified, "bioactive complex" refers to a complex containing a bioactive ingredient. A bioactive complex may be a drug-loaded complex.

[0105] In this application, unless otherwise specified, "complex" refers to a composite material assembled from multiple components and possessing a certain structural stability; non-limiting examples of complexes used herein include nanoparticles or droplets of a certain size, as well as vesicles (such as liposomes), micelles, etc.

[0106] In this application, unless otherwise specified, "bioactive preparation" refers to a preparation containing a bioactive ingredient. Bioactive preparations can be pharmaceutical preparations.

[0107] In this application, unless otherwise specified, "bioactive ingredient" refers to a substance capable of providing biological efficacy in vitro or in vivo, and often provides beneficial effects. Without limitation, biological efficacy may include functions or effects in at least one aspect of physiology and pharmacology. Exemplarily, a bioactive ingredient may include at least one of pharmaceutical ingredients, natural functional substances, etc., wherein non-limiting examples of natural functional substances include intracellular vesicles, substances contained within vesicles, etc.

[0108] In this application, unless otherwise specified, "drug ingredient" refers to a substance that can exert physiological and / or pharmacological effects, and often provides beneficial effects. The activity of "drug ingredient" is not particularly limited and can be an active substance that can interact with other substances or an inert substance that does not interact with other substances.

[0109] In this application, unless otherwise specified, "drug" includes any agent, compound, composition, or mixture that provides physiological and / or pharmacological effects in vivo or in vitro, and often provides beneficial effects. The scope of the physiological and / or pharmacological effects produced by a "drug" in vivo is not particularly limited; it may produce systemic effects or only local effects. The activity of a "drug" is not particularly limited; it may be an active substance that can interact with other substances or an inert substance that does not interact with other substances.

[0110] In this application, unless otherwise stated, "pharmaceutical active ingredient" means a pharmaceutical ingredient that can interact with other substances in vivo or in vitro.

[0111] In this application, unless otherwise specified, "pharmaceutical composition" refers to a composition containing a pharmaceutical ingredient. Generally, a pharmaceutical composition includes a pharmaceutical ingredient and a pharmaceutically acceptable carrier. Unless otherwise specified, the components of a pharmaceutical composition may together constitute a pharmaceutical preparation having a specific dosage form.

[0112] In this application, unless otherwise stated, "pharmaceutical acceptable" means suitable for administration to subjects within the scope of reasonable medical judgment and commensurate with a reasonable benefit / risk ratio, and can be used to describe ligands, materials, compositions, dosage forms, etc.

[0113] In this application, unless otherwise stated, "pharmaceuticalally acceptable carrier" means a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material.

[0114] In this application, unless otherwise stated, "drug-loaded complex" refers to a complex containing a drug component.

[0115] Unless otherwise specified in this application, "pharmaceutical preparation" refers to a preparation containing pharmaceutical ingredients that is suitable for clinical use. Pharmaceutical preparations can be used directly in clinical settings or can undergo appropriate pretreatment before clinical use, such as thawing, reconstitution, temperature adjustment (e.g., restoring to room temperature), multi-component mixing, dilution, activation of biological activity, standing, mixing (e.g., shaking), filtration, sterilization, and other pretreatment methods.

[0116] It is understood that any "pharmaceutical raw materials and excipients" mentioned in this application are pharmaceutically acceptable excipients. When implementing the technical solution of this application, those skilled in the art can make appropriate selections of relevant raw materials or components according to the administrative regulations at the time of implementation (such as excipients that are explicitly specified as usable, excipients that need to be used under restrictive conditions, and those excipients that have not been subject to any administrative regulations but have been verified to be usable), and can also exclude prohibited components according to prohibitive provisions.

[0117] In some embodiments of the first aspect of this application, an ionizable lipid compound that targets atherosclerotic cells is provided, comprising a lipid unit, an ionizable group, and a structural unit capable of targeting the function of atherosclerotic cells.

[0118] In this application, the "ionizable lipid compound targeting atherosclerosis cells" is referred to as "compound ATILipid (Atherosclerosis Targeted Inoziable Lipid)".

[0119] In this application, the "structural unit capable of targeting the function of atherosclerotic cells" can be denoted as R. 03 .

[0120] In this application, the "lipid unit" in the compound ATILipipid comprises two unsaturated fatty acyl groups, which are branched around a trivalent nitrogen (N) atom as the central branching point and connected to R. 03 Connected, the branching center of the N atom and R 03 An ionizable nitrogen atom is positioned between the two groups. Unless otherwise stated, the two unsaturated fatty acyl groups in the compound ATILibid possess anti-inflammatory activity. As a non-limiting example, the two unsaturated fatty acyl groups may each be independently derived from an unsaturated fatty acid with anti-inflammatory activity.

[0121] In this application, unless otherwise specified, the "ionizable group" in the compound ATILipid may include one or more of ionizable -NH-, ionizable -N(CH3)-, and a nitrogen-containing monocyclic ring (such as a six-membered nitrogen-containing ring) containing an ionizable nitrogen atom. In some embodiments, the ionizable group includes an ionizable N atom. In some embodiments, the ionizable group is -NH-. Exemplarily, the six-membered nitrogen-containing ring may be a nitrogen-saturated six-membered ring containing an ionizable nitrogen atom, such as a piperidine ring, a piperazine ring, etc.

[0122] In this application, "nitrogen-monocyclic ring" refers to a heterocyclic ring whose cyclic atoms include nitrogen atoms, and can be a 5- to 7-membered nitrogen-monocyclic ring. The number of cyclic atoms in the nitrogen-monocyclic ring can be 5, 6, or 7. In some embodiments, the nitrogen-monocyclic ring is a saturated ring, that is, a nitrogen-saturated monocyclic ring.

[0123] In some embodiments, the compound ATILipid includes R connected in sequence. 03 It includes ionizable N atoms and N atoms that serve as branching centers (also referred to as "N atom branching centers"), as well as two unsaturated fatty acyl groups connected to the N atom branching centers.

[0124] In some embodiments, the compound ATILibid sequentially comprises two anti-inflammatory unsaturated fatty acyl groups linked to a branched N atom via a divalent linker, the branched N atom, a divalent linker G containing an ionizable N atom, and a structural unit R capable of targeting atherosclerotic cell function. 03 Furthermore, the two unsaturated fatty acyl groups can be connected to the N-atom branching center via divalent linkers Z1-L1 and Z2-L2, respectively. In some embodiments, the two unsaturated fatty acyl groups can be connected to the N-atom branching center via amide bonds and carbon-carbon chains, respectively; furthermore, the carbon-carbon chain can be an alkylene group, more particularly an alkylene group with 1 to 3 carbon atoms in the main chain, and even more particularly a C-carbon chain. 1-3 Alkylene.

[0125] In some embodiments of the first aspect of this application, an ionizable lipid compound (compound ATILibid) targeting atherosclerotic cells is provided, which is a compound with the structure shown in formula (I);

[0126]

[0127] In formula (I),

[0128] R1- and R2- each independently form an unsaturated fatty acyl group with anti-inflammatory activity with the carbonyl group;

[0129] Z1 and Z2 are each independently O, NH or S;

[0130] L1 and L2 are each independently an alkylene group with 1 to 3 carbon atoms in the main chain;

[0131] G is a divalent aliphatic hydrocarbon group containing one or more ionizable N atoms;

[0132] R3 includes structural units R that can target the function of atherosclerotic cells. 03 .

[0133] In some other embodiments of the first aspect of this application, an ionizable lipid compound that targets atherosclerotic cells is also provided, which is a compound with the structure shown in formula (I);

[0134] In formula (I), R1- and R2- each independently form an unsaturated fatty acyl group with the carbonyl group derived from α-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, medelic acid or sinapic acid.

[0135] Z1 and Z2 are each independently O, NH or S;

[0136] L1 and L2 are each independently C 1-3 Alkylene;

[0137] G is a divalent aliphatic hydrocarbon group containing one or more ionizable N atoms;

[0138] R3 includes structural units R that have the function of targeting atherosclerotic cells. 03 .

[0139] The ionizable lipid compound ATILipid can target atherosclerotic cells and has anti-inflammatory activity. It can be used as an anti-inflammatory drug to improve plaque inflammation, and it can also serve as a structural component of bioactive complexes such as lipid nanoparticles (e.g., drug complexes). It can co-deliver bioactive components such as small molecules, peptides, and gene drugs, and can be used for synergistic treatment of atherosclerosis, but is not limited to this application.

[0140] This ionizable lipid compound can promote the targeted accumulation of the encapsulated bioactive components in atherosclerotic plaques.

[0141] The aforementioned ionizable lipid compound ATILibid, which targets atherosclerotic cells, can be applied in the treatment of atherosclerosis and is of significant importance in promoting the cure of cardiovascular and other chronic inflammatory diseases. On the one hand, compared to traditional ionizable lipid molecules, ATILibid has anti-inflammatory effects, improving inflammatory responses at lesion sites and targeting and accumulating at the lesion site to achieve precise treatment. On the other hand, compared to co-delivering anti-inflammatory drugs, ATILibid can serve as a component of drug delivery systems such as liposomes, lipid nanoparticles, and polymer micelles, directly participating in the structural formation of drug-loaded complexes. This can enhance nucleic acid transfection and drug delivery effects while mitigating, to some extent, the unknown risks inherent in drug development.

[0142] This ionizable lipid compound can be used for the treatment of atherosclerotic inflammation.

[0143] The ionizable lipid compound ATILibid can be used for targeted therapy of inflammatory diseases. ATILibid comprises monounsaturated or polyunsaturated fatty acid tail chains with anti-inflammatory activity, which can inhibit the secretion of inflammatory factors by cells, thereby reducing the inflammatory response at the disease site. ATILibid also has a head group targeting atherosclerosis, which can increase the targeted accumulation of lipids at the disease site, improve drug delivery via drug delivery carriers or drug-loaded complexes, enhance the accumulation of the drug-loaded complex itself at the lesion site, and strengthen both the efficacy of the drug and the anti-inflammatory effect of the drug-loaded complex.

[0144] In some embodiments, R1- and R2- each independently form an unsaturated fatty acyl group with anti-inflammatory activity with the carbonyl group.

[0145] In this application, unless otherwise specified, "unsaturated fatty acyl group" refers to a carbon acyl group containing an unsaturated fatty chain. Unless otherwise specified, an unsaturated fatty acyl group is composed of an unsaturated fatty chain and a -C (=O)- group located at the linking site. Typically, the unsaturated fatty chain is an unsaturated hydrocarbon group containing an unsaturated carbon-carbon bond. In some embodiments, the unsaturated carbon-carbon bond is a carbon-carbon double bond. The unsaturated fatty acyl group can be the dehydroxylated portion of an unsaturated fatty acid. In some embodiments, the number of carbon atoms in the unsaturated fatty acyl group can be 18 to 22, for example, 18, 20, or 22.

[0146] In some implementations, R1-C(=O)- and R2-C(=O)- each independently contain 18 to 22 carbon atoms, for example, each can independently contain 18, 20 or 22 carbon atoms.

[0147] In some embodiments, R1-C(=O)- and R2-C(=O)- each independently contain 1 to 6 carbon-carbon double bonds, for example, they may contain 1, 2, 3, 4, 5 or 6 carbon-carbon double bonds. Non-limitingly, the number of carbon-carbon double bonds in R1-C(=O)- and R2-C(=O)- can also be selected from any two integers from 1, 2, 3, 4, 5 and 6.

[0148] In some embodiments, the anti-inflammatory unsaturated fatty acyl group is derived from an anti-inflammatory unsaturated fatty acid. Those anti-inflammatory unsaturated fatty acids known in the art that are biocompatible can be selected, and these can be monounsaturated or polyunsaturated fatty acids with anti-inflammatory activity. Non-limiting examples include ω-3 polyunsaturated fatty acids (such as eicosapentaenoic acid EPA, docosapentaenoic acid DPA, docosahexaenoic acid DHA, alpha-linolenic acid (ALA), etc.), mideic acid, sinapic acid, etc.

[0149] In some embodiments, R1- and R2- each independently form an unsaturated fatty acyl group derived from an ω-3 polyunsaturated fatty acid with the carbonyl group.

[0150] In some embodiments, R1- and R2- each independently form an unsaturated fatty acyl group with the carbonyl group derived from α-linolenic acid, eicosapentaenoic acid (such as EPA), docosapentaenoic acid (such as DPA), docosahexaenoic acid (such as DHA), medelic acid, or sinapic acid.

[0151] In some embodiments, the unsaturated fatty acid with anti-inflammatory activity is an ω-3 polyunsaturated fatty acid, which may further be EPA or DHA.

[0152] In some implementations, the unsaturated fatty acid with anti-inflammatory activity is EPA.

[0153] The structure of alpha-linolenic acid (ALA) is:

[0154] The structure of EPA is

[0155] The structure of DPA is

[0156] The structure of DHA is

[0157] Non-limiting examples of monounsaturated or polyunsaturated fatty acids with anti-inflammatory activity include:

[0158]

[0159] In some implementations, R 01 -and R 02-Each group independently forms one of the following acyl groups with a carbonyl group:

[0160]

[0161] In some embodiments, R1-C(=O)- and R2-C(=O)- are each independently a fatty acyl group derived from eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA).

[0162] In some implementations, Z1 and Z2 are each independently O, NH or S.

[0163] In some implementations, Z1 and Z2 are each independently zero.

[0164] In some embodiments, L1 and L2 are each independently an alkylene group having 1 to 3 carbon atoms in the main chain.

[0165] In this application, unless otherwise specified, "alkylene with 1 to 3 carbon atoms in the main chain" refers to a divalent alkyl group having a "*-(C)n-*" skeleton, where n is an integer selected from 1 to 3, such as 1, 2 or 3.

[0166] In some implementations, L1 and L2 are each independently -(CR a R b ) q -, where R a and R b Each is independently a H atom or a methyl group, and q is an integer selected from 1 to 3. q can be 1, 2, or 3, and more specifically, 2. In some embodiments, R... a and R b All are H atoms.

[0167] In some implementations, L1 and L2 are each independently C 1-3 The alkylene group can further be methylene, ethylene, or propylene, and even further can be methylene, 1,2-ethylene, or 1,3-propylene.

[0168] In some implementations, L1 and L2 are each independently -(CH2). p - where p is an integer selected from 1 to 3; further, p can be 1, 2 or 3.

[0169] In some implementations, L1 and L2 are each independently -CH2CH2-.

[0170] In some embodiments, G is a divalent aliphatic hydrocarbon group containing one or more ionizable N atoms. Further, G is a divalent aliphatic hydrocarbon group whose main chain atoms include one or more ionizable N atoms, meaning one or more ionizable N atoms are located in the main chain of the divalent aliphatic hydrocarbon group.

[0171] In this application, unless otherwise specified, "divalent aliphatic hydrocarbon group" refers to an aliphatic divalent hydrocarbon group containing heteroatoms. As used herein, "heteroatom" refers to a type of atom other than hydrogen and carbon atoms, capable of participating in the formation of covalent bonds in compounds.

[0172] In some embodiments, G includes a divalent group G0, wherein G0 is composed of one or more straight-chain saturated alkylene groups and one or more ionizable groups. The ionizable groups may include ionizable nitrogen-containing groups.

[0173] In some embodiments, the ionizable group is an ionizable nitrogen-containing group.

[0174] Without limitation, the ionizable nitrogen-containing group may include at least one of ionizable -NH-, ionizable -N(CH3)-, and a nitrogen-containing monocyclic ring containing an ionizable nitrogen atom.

[0175] In some embodiments, the ionizable group is selected from at least one of ionizable -NH-, ionizable -N(CH3)-, and a nitrogen-containing monocyclic ring.

[0176] In some embodiments, the nitrogen heterocyclic ring containing ionizable nitrogen atoms can be a six-membered nitrogen heterocyclic ring containing ionizable nitrogen atoms.

[0177] In some embodiments, the six-membered nitrogen heterocycle containing ionizable nitrogen atoms can be a nitrogen-saturated six-membered ring containing ionizable nitrogen atoms.

[0178] In some embodiments, the number of nitrogen atoms in the nitrogen-containing heterocyclic ring is 1 or 2.

[0179] In some embodiments, the nitrogen-containing monocyclic ring with ionizable nitrogen atoms is selected from at least one of piperidine ring and piperazine ring.

[0180] In some embodiments, the piperidine ring is (1,4-piperidine ring), piperazine ring is (1,4-piperazine ring).

[0181] In some embodiments, G0 is composed of one or more straight-chain saturated alkylene groups and one or more ionizable nitrogen-containing groups.

[0182] In some embodiments, G0 consists of one or two straight-chain saturated alkylene groups and an ionizable nitrogen-containing group.

[0183] In some embodiments, the straight-chain saturated alkylene group is C 2-3 The straight-chain saturated alkylene group may further be 1,2-ethylene or 1,3-propylene.

[0184] In some embodiments, G0 is selected from at least one of the following divalent groups: Alternatively, G0 is Unless otherwise specified, either end of the aforementioned divalent group may point to R3. In some embodiments, the terminal carbon atom of the aforementioned divalent group points to the branching center of the N atom. In some embodiments, the terminal N atom of the aforementioned divalent group points to R3.

[0185] In some embodiments, the structural unit R that can target the function of atherosclerotic cells 03 It includes at least one of the functional residues of the following substances: small molecule organic compounds and polypeptides or proteins having at least 4 amino acid units.

[0186] In some implementations, R 03 It includes at least one of the functional residues of the following substances: polypeptides, monoclonal antibodies, and carbohydrate molecules.

[0187] Non-limiting examples of monoclonal antibodies include at least one of ASA6 monoclonal antibody, Anti-CD47 antibody, and OPN antibody (osteopontin antibody).

[0188] Non-limiting examples of carbohydrate molecules include at least one of hyaluronic acid, chitosan, and mannose.

[0189] In some implementations, R 03 Including polypeptide structural units and At least one of them;

[0190] In some implementations, R 03 Includes targeting peptide units.

[0191] In some implementations, R 03 This includes targeting peptide units based on one or more of the following polypeptides: VCAM-1 targeting peptide, ICAM-1 targeting peptide, P-selectin targeting peptide, S2P targeting peptide, IL-4 receptor targeting peptide, MCP-1 targeting peptide, CCR2 targeting peptide, collagen targeting peptide, CD44 targeting peptide, CD36 targeting peptide, SR-B1 targeting peptide, LOX-1 targeting peptide, fibrin targeting peptide, matrix metalloproteinase targeting peptide, integrin αvβ3 targeting peptide, oxidized low-density lipoprotein targeting peptide, and inflammasome targeting peptide.

[0192] In some implementations, R 03 Includes VCAM-1 targeting peptide units.

[0193] The targeting peptide unit can be a chain peptide, a cyclic peptide, or a combination of chain and cyclic peptides. Non-limitingly, the amino acid units in the chain peptide are sequentially linked by peptide bonds to form a chain structure. Non-limiting examples of the amino acid sequence of the chain peptide may include one or more of SEQ ID No. 2, SEQ ID No. 4, SEQ ID No. 5, etc. Non-limiting examples of the amino acid sequence of the cyclic peptide may include one or more of SEQ ID No. 3, SEQ ID No. 6, SEQ ID No. 7, etc.

[0194] In some implementations, R 03 This includes a VCAM-1 targeting peptide unit containing a VHPK peptide (SEQ ID No. 1), or R3 is any of the following polypeptide structural units: VHPKQHRC (SEQ ID No. 2), cyclic CNNSKSHTC (cyclic) (SEQ ID No. 3), VHPKQHRGGSGC (SEQ ID No. 4), VHPKQHRGGSKGC (SEQ ID No. 5), cyclic FLDRK (cyclic) (SEQ ID No. 6), and cyclic NNSKSHT (cyclic) (SEQ ID No. 7); wherein the (cyclic) in the cyclic polypeptide structural unit indicates that the amino acid units at both ends are linked by peptide bonds to form a ring. For example, "cyclic CNNSKSHTC (cyclic)" means that the two amino acid units C (cysteine) at both ends are linked by peptide bonds to form a ring.

[0195] Table 1.

[0196]

[0197] In this application, the amino acid sequence of the peptide, unless otherwise specified, can refer to either C-terminus to N-terminus or N-terminus to C-terminus.

[0198] In some embodiments, the peptide involved in R3 is a side chain (such as a side thiol group) of a C-terminal amino acid linked to G in Formula I.

[0199] In other embodiments, the peptide involved in R3 is a side chain of an N-terminal amino acid or an N-terminal amino group linked to G in Formula I.

[0200] In this application, cyclic peptide segments are involved, which can be represented by "X1..X2" to indicate that the X1st and X2th amino acids in the amino acid sequence are linked together. The linkage can be via peptide bonds or via the side chains of amino acids.

[0201] In some embodiments, the two cysteine ​​units in CNNSKSHTC(cyclic) (SEQ ID No. 3) form a ring by a disulfide bond through a peptide bond, and the side thiol group of the cysteine ​​at position 9 is connected via... It forms a covalent connection with G.

[0202] In some implementations, the cyclic positions of the cyclic peptide units in Table 1 are linked intrachain via peptide bonds.

[0203] In some embodiments, G and R3 are linked by at least one of the following structures: chemical bond, -NH-, -CH2NH-*, *-C(=O)NH-, *-NHC(=O)-, -SS-, *-C(=O)S-, *-SC(=O)-, *-OC(=O)-, or *-C(=O)O-; wherein * is near R3 and the other end is near G. Exemplarily, -S- can be a residue left by the reaction of a cysteine ​​side thiol group (-SH); -C(=O)- can be a residue left by the reaction of a C-terminal carboxyl group or a carboxyl group located on a side group of the peptide; -NH- can be a residue left by the reaction of an N-terminal amino group or an amino group located on a side group of the peptide; and -O- can be a residue left by the reaction of a side hydroxyl group of the peptide.

[0204] In some embodiments, G is selected from any of the following groups:

[0205] * indicates a covalently coupled site connecting to R3.

[0206] In some embodiments, R3-G- is selected from any of the following groups:

[0207]

[0208] In some embodiments, the ionizable lipid compound ATILipid has the structure shown in formula (III):

[0209]

[0210] In some embodiments, the ionizable lipid compound ATILibid is any one of the compounds in Table 2.

[0211] Table 2.

[0212]

[0213] In some embodiments, the ionizable lipid compound ATILibid is the compound shown in formula (IV):

[0214]

[0215] In some embodiments of the second aspect of this application, a delivery carrier composition is provided comprising an ionizable lipid compound targeting atherosclerotic cells as described in the first aspect of this application.

[0216] In some embodiments, the delivery carrier composition includes an ionizable lipid compound, an optional steroid, an optional auxiliary lipid, an optional PEGylated lipid, and optional other pharmaceutical excipients.

[0217] In some embodiments, the delivery carrier composition comprises an ionizable lipid compound, an optional steroid, an optional auxiliary lipid, an optional PEGylated lipid, and optional other pharmaceutical excipients.

[0218] In some embodiments, the delivery carrier composition includes at least one of a steroid, an auxiliary lipid, and a polyethylene glycol-modified lipid.

[0219] In some embodiments, the delivery carrier composition includes at least two of steroids, auxiliary lipids, and PEGylated lipids.

[0220] In some embodiments, the delivery carrier composition includes a steroid.

[0221] In some embodiments, the delivery carrier composition includes auxiliary lipids.

[0222] In some embodiments, the delivery carrier composition comprises polyethylene glycol-modified lipids.

[0223] In some embodiments, the delivery carrier composition includes steroids, auxiliary lipids, and polyethylene glycol-modified lipids.

[0224] In some embodiments, a delivery carrier composition is provided, comprising, by molar percentage, 20% to 70% of an ionizable lipid compound, 0% to 60% of a steroid, 0% to 30% of an auxiliary lipid, 0% to 10% of a polyethylene glycol-modified lipid, and 0% to 15% of other pharmaceutical excipients; or, by mass percentage, the delivery carrier composition comprises 20% to 90% of an ionizable lipid compound, 0% to 50% of a steroid, 0% to 50% of an auxiliary lipid, 0% to 20% of a polyethylene glycol-modified lipid, and 0% to 80% of other pharmaceutical excipients; wherein the ionizable lipid compound is at least one of the ionizable lipid compounds (ATILipid) targeting atherosclerotic cells described in the first aspect of this application.

[0225] It is understandable that other pharmaceutical excipients differ from any of the following: ionizable lipid compounds, steroids, auxiliary lipids, and PEGylated lipids.

[0226] In some embodiments, the delivery carrier composition may contain any two of the following: an ionizable lipid compound, a steroid, an auxiliary lipid, a PEGylated lipid, and other pharmaceutical excipients.

[0227] In some embodiments, in the delivery carrier composition, the polyethylene glycolated lipid is different from any of the ionizable lipid compounds, steroids, and auxiliary lipids; the steroid is different from any of the auxiliary lipids and the polyethylene glycolated lipids.

[0228] In some embodiments, the auxiliary lipids include one or more of phosphatidylethanolamines, phosphatidylcholine derivatives, betaine lipids (DGTS), and sphingomyelins (SM). Non-limiting examples of phosphatidylethanolamines may include dicofol phosphatidylethanolamine. Non-limiting examples of phosphatidylcholine derivatives may include dicofol phosphatidylcholine.

[0229] Non-limitingly, the molar percentage of the ionizable lipid compound ATILibid in the delivery carrier composition may be 20% to 70%, optionally 30% to 70%, further optionally 30% to 60%, and may also be any of the following percentages or a range selected from any two of the following percentages: 20%, 25%, 30%, 35%, 40%, 50%, 60%, 65%, 70%, etc.

[0230] Non-limitingly, the molar percentage of steroid in the delivery carrier composition can be 0% to 60%, optionally 5% to 60%, further optionally 15% to 45%, and can also be any of the following percentages or a range selected from any two of the following percentages: 0%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, etc.

[0231] Non-limitingly, the molar percentage of the auxiliary lipid in the delivery carrier composition can be 0% to 30%, optionally 5% to 30%, further optionally 5% to 20%, and can also be any of the following percentages or a range selected from any two of the following percentages: 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 15%, 16%, 18%, 20%, 25%, 30%, etc.

[0232] Non-limitingly, the molar percentage of polyethylene glycol-modified lipids in the delivery carrier composition may be 0% to 10%, optionally 1% to 5%, or any of the following percentages or a range selected from any two of the following percentages: 0%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, etc.

[0233] Non-limitingly, the molar percentage of other pharmaceutical excipients in the delivery carrier composition may be 0% to 15%, optionally 5% to 10%, or any of the following percentages or a range selected from any two of the following percentages: 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 15%, etc.

[0234] Non-limitingly, the mass percentage of the ionizable lipid compound ATILipid in the delivery carrier composition may be 20% to 90%, optionally 30% to 90%, further optionally 40% to 90%, and may also be any of the following percentages or a range selected from any two of the following percentages: 20%, 25%, 30%, 35%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, etc.

[0235] In some embodiments, the sum of the mass percentages of the steroid, the auxiliary lipid, and the polyethylene glycol-modified lipid in the delivery carrier composition is 20% to 80%, optionally 30% to 80%, or 20% to 75%, or any of the following percentages or a range selected from any two of the following percentages: 20%, 25%, 30%, 35%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, etc.

[0236] Non-limitingly, the steroid in the delivery carrier composition may be 0% to 50% by mass, optionally 5% to 50%, further optionally 5% to 45%, and may also be any of the following percentages or a range selected from any two of the following percentages: 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 15%, 16%, 18%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, etc.

[0237] Non-limitingly, the mass percentage of the auxiliary lipid in the delivery carrier composition can be 0% to 50%, optionally 0% to 30% or 5% to 50%, and can also be any of the following percentages or a range selected from any two of the following percentages: 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 15%, 16%, 18%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, etc.

[0238] Non-limitingly, the mass percentage of polyethylene glycol-modified lipids in the delivery carrier composition can be 0% to 20%, optionally 0% to 10% or 2% to 20%, and can also be any of the following percentages or a range selected from any two of the following percentages: 0%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 15%, 16%, 18%, 20%, etc.

[0239] Non-limitingly, the mass percentage of other pharmaceutical excipients in the delivery carrier composition may be 0% to 80%, optionally 10% to 60%, or any of the following percentages or a range selected from any two of the following percentages: 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, etc.

[0240] Without limitation, steroids may include one or more of the following substances: cholesterol, lanosterol, sitosterol, stigmasterol, ergosterol, sulfasalazine, campesterol, and brassicasterol. In some embodiments, steroids include cholesterol.

[0241] In some implementations, the auxiliary lipid is a neutral lipid.

[0242] Non-limitingly, the cofactor lipid may include at least one of distearylphosphatidylcholine (DSPC), distearylphosphatidylethanolamine (DSPE), dioleoylphosphatidylethanolamine (DOPE), dioleoyllecithin (DOPC), dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylcholine (DMPC), dimyristoylphosphatidylethanolamine (DMPE), 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine (POPC), dioleoylphosphatidylglycerol (DOPG), betaine lipids (DGTS), and sphingomyelin (SM). In some embodiments, the cofactor lipid includes one or more of DSPC, DSPE, DOPE, and DOPC.

[0243] Without limitation, the lipids in the polyethylene glycol-modified lipids may include one or more of dimyristoyl glycerol (DMG), distearylphosphatidylethanolamine (DSPE), distearyl glycerol (DSG), dipalmitoyl glycerol, dioleoyl glycerol, distearyl glycerol, diacylglycamide (DAG), dipalmitoylphosphatidylethanolamine (DPPE), polyethyleneimine (PE), succinoyl-diacylglycerol (S-DAG), dialkoxypropyl carbamate, and dimethacrylate (c-DMA).

[0244] Non-limitingly, the weight-average molecular weight of the polyethylene glycol portion in the polyethylene glycol-modified lipid can be 1500 Da to 2500 Da, with non-limiting examples such as 1500 Da, 1600 Da, 1700 Da, 1800 Da, 1900 Da, 2000 Da, 2100 Da, 2200 Da, 2300 Da, 2400 Da, 2500 Da, etc., and can also be selected from any of the above two molecular weight ranges.

[0245] In some embodiments, the polyethylene glycol portion of the PEGylated lipoprotein includes polyethylene glycol monomethyl ether, polyethylene glycol hydroxyl, or polyethylene glycol amino. Optionally, the polyethylene glycol portion of the PEGylated lipoprotein is polyethylene glycol monomethyl ether, polyethylene glycol hydroxyl, or polyethylene glycol amino.

[0246] Without limitation, PEGylated lipids may include one or more of the following: methoxy polyethylene glycol bis(tetradecyl acetamide) (ALC-0159), PEG-DMG (polyethylene glycol dimyristoyl glycerol), PEG-DSPE (polyethylene glycol distearate phosphatidylethanolamine), PEG-DSG (polyethylene glycol distearate glycerol), PEG-DPG (polyethylene glycol dipalmitoyl glycerol), PEG-DOPE (polyethylene glycol dioleoyl phosphatidylethanolamine), PEG-distearate, PEG-DAG (polyethylene glycol diacylglycamide), PEG-DPPE (polyethylene glycol dipalmitoyl phosphatidylethanolamine), PEG-PE (polyethylene glycol polyethyleneimine), PEG-S-DAG (polyethylene glycol succinoyl-diacylglycerol), PEG-dialkoxypropylcarbamate, and PEG-c-DMA (polyethylene glycol dimethacrylate). In some embodiments, PEGylated lipids may include one or more of DMG-PEG2000, DSPE-MPEG2000, ALC-0159, and PEG-DOPE.

[0247] In some embodiments, the delivery carrier composition may optionally include other pharmaceutical excipients. Non-limitingly, other pharmaceutical excipients may be selected from one or more of the following pharmaceutical excipients: surfactants, emulsifiers, co-emulsifiers, solubilizers, buffers, osmotic pressure regulators, antibacterial agents, and protein stabilizers. Non-limiting examples of co-emulsifiers may include oleic acid. Non-limiting examples of surfactants may include Tween 80. Non-limiting examples of osmotic pressure regulators may include glycerol. In some embodiments, other pharmaceutical excipients include one or more of surfactants and osmotic pressure regulators.

[0248] In some embodiments, the delivery carrier composition constitutes a drug delivery carrier. Without limitation, the drug delivery carrier may be a liposome, lipid nanoparticle, polymer nanoparticle, nanomicelle, or nanoemulsion drug carrier.

[0249] In some embodiments of another aspect of this application, a drug delivery carrier is provided, comprising the delivery carrier composition described in the second aspect of this application.

[0250] In some embodiments, the drug delivery carrier is a liposome, lipid nanoparticle, polymer nanoparticle, nanomicelle, or nanoemulsion drug carrier.

[0251] In this application, unless otherwise specified, "liposome" refers to a synthetically produced vesicle structure comprising a lipid bilayer and an internal cavity formed by the bilayer. Typically, the inner surface of the bilayer is hydrophilic, allowing the internal cavity to contain hydrophilic substances. The lipid components in liposomes, including but not limited to phospholipids, can be of synthetic or natural origin.

[0252] In this application, unless otherwise stated, "lipid nanoparticles" refers to nanoparticles formed by the assembly of lipid components.

[0253] In this application, unless otherwise stated, "polymer nanoparticles" refers to nanoparticles formed by the assembly of polymers.

[0254] In this application, unless otherwise stated, "micelle" refers to an aggregate formed by the assembly of amphiphilic molecules. Unless otherwise stated, the outer surface of a micelle is hydrophilic, and the interior of a micelle may contain hydrophobic substances.

[0255] Unless otherwise stated in this application, "nanoemulsion drug carrier" refers to an emulsion-like drug carrier in which the droplets are nanoscale.

[0256] Unless otherwise stated in this application, the "nano" scale is in the range of 1 nm to 1000 nm.

[0257] In some embodiments, the average particle size of the drug delivery carrier is 50 nm to 200 nm, optionally 50 nm to 100 nm, or any of the following particle sizes or a range selected from any two of the following particle sizes: 50 nm, 60 nm, 80 nm, 100 nm, 120 nm, 150 nm, 160 nm, 180 nm, 200 nm, etc.

[0258] In some embodiments of the third aspect of this application, a bioactive composition, bioactive complex, or bioactive formulation is provided, wherein the bioactive formulation includes a bioactive complex; the bioactive composition or bioactive complex independently includes a bioactive ingredient and an ionizable lipid compound (ATILipid) targeting atherosclerotic cells as described in the first aspect of this application.

[0259] In some embodiments, the bioactive ingredient includes a pharmaceutical ingredient, the bioactive composition is a pharmaceutical composition, the bioactive complex is a drug-loaded complex, and the bioactive preparation is a pharmaceutical preparation.

[0260] In some implementations, the bioactive ingredient includes a pharmaceutically active ingredient.

[0261] By loading bioactive components onto the aforementioned ionizable lipid compound ATILipid with anti-inflammatory activity and drug delivery carriers (such as liposomes, lipid nanoparticles, and other complexes), the resulting drug-loaded complexes or drug formulations can deliver bioactive components into the body, which is beneficial for the loaded or delivered bioactive components to exert their effects better. This can play an advantage in the targeted treatment of immune-related inflammatory cardiovascular diseases.

[0262] The aforementioned ionizable lipid compound ATILibid can be used as a component of pharmaceutical compositions to prepare drug formulations. Its ionization properties allow for the delivery of bioactive therapeutic drugs such as nucleic acids, improving the safety and transfection efficiency of nucleic acid drugs in vivo. Exemplary drug-loaded complexes or drug formulations may include lipid nanoparticles, liposomes, and lipid nanovesicles. Pharmaceutical compositions, drug-loaded complexes, and drug formulations incorporating the ionizable lipid compound ATILibid offer advantages such as inhibiting inflammatory responses, promoting targeted drug aggregation at lesions, and achieving high nucleic acid transfection efficiency.

[0263] In some other embodiments of the third aspect of this application, a pharmaceutical composition, a drug-loaded complex, or a pharmaceutical formulation is provided, the pharmaceutical formulation including the drug-loaded complex; the pharmaceutical composition or drug-loaded complex independently includes a bioactive ingredient and an ionizable lipid compound (ATILipid) targeting atherosclerotic cells as described in the first aspect of this application.

[0264] In some embodiments, the bioactive formulation includes a bioactive ingredient and the aforementioned delivery carrier composition.

[0265] In some embodiments, the pharmaceutical formulation includes a bioactive ingredient and the aforementioned delivery carrier composition.

[0266] In some embodiments, the bioactive composition (such as a pharmaceutical composition) or bioactive complex (such as a drug-loaded complex) also independently includes at least one of steroids, cofactor lipids, and PEGylated lipids. Non-limitingly, the cofactor lipid may include one or more of phosphatidylethanolamines (non-limiting examples may include dicofol), phosphatidylcholine derivatives (non-limiting examples may include dicofol), betaine lipids (DGTS), and sphingomyelin (SM).

[0267] In some embodiments, the average particle size of the bioactive complex (such as a drug-loaded complex) is 50 nm to 200 nm, optionally 50 nm to 100 nm, or any of the following particle sizes or a range selected from any two of the following particle sizes: 50 nm, 60 nm, 80 nm, 100 nm, 120 nm, 150 nm, 160 nm, 180 nm, 200 nm, etc.

[0268] In some embodiments, the bioactive complex is a drug-loaded complex. Without limitation, the drug-loaded complex may be a drug-loaded liposome, drug-loaded lipid nanoparticle, drug-loaded polymer nanoparticle, drug-loaded nanomicelles, or nanoemulsion drug formulation.

[0269] In some embodiments, the drug-loaded complex or pharmaceutical formulation includes a bioactive ingredient and a drug delivery carrier, wherein the bioactive ingredient is loaded in the drug delivery carrier; further, when the drug-loaded complex has a nanoscale size, the drug-loaded complex is also referred to as a drug nanocarrier complex.

[0270] In some embodiments, the pharmaceutical preparation is a drug-loaded lipid preparation.

[0271] Without limitation, bioactive ingredients may include at least one of nucleic acid substances, proteins, peptides, and small molecule organic drugs.

[0272] In some implementations, the bioactive ingredient carries a negative charge or is a hydrophobic substance.

[0273] In some embodiments, the bioactive ingredient includes nucleic acid-like substances. Without limitation, nucleic acid-like substances may include one or more of miRNA, mRNA, siRNA, ASO, saRNA, shRNA, Pre-RNA, lncRNA, RNA mimics, RNAagomir, RNA antagomir, DNA, and cDNA (complementary DNA).

[0274] In some embodiments, the mass ratio of the bioactive ingredient to the ionizable lipid compound is 1:(0.5 to 1000), optionally 1:(0.5 to 200), further optionally 1:(1 to 100), and may also be any of the following ratios or a range selected from any two of the following ratios: 1:0.5, 1:0.6, 1:0.7, 1:0.75, 1:0.8, 1:0.9, 1:1, 1:1.25, 1:1.5, 1:1.75, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:7.5, 1:8, 1:9, 1 :10, 1:12, 1:15, 1:16, 1:18, 1:20, 1:22, 1:24, 1:25, 1:26, 1:28, 1:30, 1:35, 1:40, 1:45, 1:50, 1:60, 1:70, 1:80, 1:90, 1:100, 1:120, 1:140, 1:150, 1:160, 1:180, 1:200, 1:250, 1:300, 1:350, 1:400, 1:450, 1:500, 1:600, 1:700, 1:800, 1:900, 1:1000, etc. Without limitation, the mass ratio of the bioactive component to the ionizable lipid compound may also be selected from any of the following ratios: 1:(0.5–100), 1:(0.5–30), 1:(0.5–20), 1:(1–200), 1:(1–100), 1:(1–50), 1:(1–20), etc.

[0275] In some embodiments, the bioactive ingredient includes nucleic acid-like substances, wherein the mass ratio of the nucleic acid-like substances to the ionizable lipid compound is 1:(1-1000), optionally 1:(1-200), further optionally 1:(1-200), and even more preferably 1:(1-100). It can also be any of the following ratios or a range selected from any two of the following ratios: 1:1, 1:1.25, 1:1.5, 1:1.75, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:7.5, 1:8, 1:9, 1:10, 1... :12, 1:15, 1:16, 1:18, 1:20, 1:22, 1:24, 1:25, 1:26, 1:28, 1:30, 1:35, 1:40, 1:45, 1:50, 1:60, 1:70, 1:80, 1:90, 1:100, 1:120, 1:140, 1:150, 1:160, 1:180, 1:200, 1:250, 1:300, 1:350, 1:400, 1:450, 1:500, 1:600, 1:700, 1:800, 1:900, 1:1000, etc. Without limitation, the mass ratio of nucleic acid substances to ionizable lipid compounds may also be selected from any of the following ratios: 1:(1-50), 1:(1-30), 1:(1-20), etc.

[0276] In some embodiments, the bioactive ingredient includes nucleic acid-like substances, wherein the mass ratio of the nucleic acid-like substances to the ionizable lipid compound is 1:(1-1000), optionally 1:(1-200), further optionally 1:(1-100), and may also be any of the following ratios or a range selected from any two of the following ratios: 1:10, 1:12, 1:15, 1:16, 1:18, 1:20, 1:22, 1:24, 1:25, 1:26, 1:28, 1:30, 1 :35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, 1:95, 1:100, 1:120, 1:140, 1:150, 1:160, 1:180, 1:200, 1:250, 1:300, 1:350, 1:400, 1:450, 1:500, 1:600, 1:700, 1:800, 1:900, 1:1000, etc. Non-limitingly, the mass ratio of nucleic acid substances to ionizable lipid compounds can also be selected from any of the following ratios: 1:(10–1000), 1:(10–200), 1:(10–100), etc.

[0277] In some embodiments, the bioactive composition (such as a pharmaceutical composition) or bioactive formulation (such as a pharmaceutical formulation) independently includes the delivery carrier composition described in the second aspect of this application. Without limitation, the bioactive ingredient may be a nucleic acid-based substance. In some embodiments, the delivery carrier composition comprises, by weight percentage, 50%–70% of an ionizable lipid compound, 25%–35% of a steroid, 5%–30% of an auxiliary lipid, 1%–5% of a polyethylene glycol-modified lipid, and 5%–10% of other pharmaceutical excipients; further, the mass ratio of the nucleic acid-based substance to the ionizable lipid compound is 1:(10–100), or may be any of the following ratios. Or selected from any two of the following ratios: 1:10, 1:12, 1:15, 1:16, 1:18, 1:20, 1:22, 1:24, 1:25, 1:26, 1:28, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, 1:95, 1:100, etc.

[0278] In some embodiments, the bioactive preparation or pharmaceutical preparation is an injection.

[0279] In some embodiments, the bioactive formulation (such as a pharmaceutical formulation) is a liquid formulation, and the solvent includes water. Non-limitingly, the mass percentage of the bioactive complex (such as a drug-loaded complex) in the liquid formulation can be 1% to 20%, and can also be any of the following percentages or a range selected from any two of the following percentages: 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 15%, 16%, 18%, 20%, etc.

[0280] In some embodiments of the fourth aspect of this application, the use of the ionizable lipid compound (ATILipid) targeting atherosclerotic cells described in the first aspect of this application is provided in the preparation of drug delivery carriers, bioactive complexes or drug-loaded complexes, or in the preparation of medicaments for treating diseases. Alternatively, the use of the ionizable lipid compound (ATILipid) targeting atherosclerotic cells described in the first aspect of this application, the delivery carrier composition described in the second aspect of this application, the drug delivery carrier (such as liposomes or lipid nanoparticles) described in one aspect of this application, or the bioactive composition or bioactive complex described in the third aspect of this application, or the drug composition or drug-loaded complex described in the third aspect of this application, in the preparation of pharmaceutical formulations for treating diseases or in the preparation of medicaments for treating diseases is provided.

[0281] Non-limitingly, the treatment of diseases may include at least one of the following: treatment of chronic inflammatory diseases, targeted therapy for inflammatory diseases, targeted therapy for atherosclerosis, treatment of atherosclerotic inflammation, treatment of familial hyperlipidemia, treatment of atherosclerotic lipid deposition, treatment of hypertension caused by atherosclerosis, treatment of atherosclerotic stroke, and treatment of atherosclerotic vascular damage. Non-limitingly, inflammatory diseases may include plaque inflammation. Non-limitingly, chronic inflammatory diseases may include cardiovascular diseases.

[0282] In some implementations, the disease includes at least one of immune-related inflammatory cardiovascular disease, atherosclerotic inflammation, atherosclerotic myocardial infarction, and atherosclerotic stroke.

[0283] In some embodiments of another aspect of this application, the use of the ionizable lipid compound (ATILipid) targeting atherosclerotic cells described in the first aspect of this application is provided in the preparation of liposomes, lipid nanoparticles, polymer nanoparticles, nanomicelles, or nanoemulsion drug carriers.

[0284] In some embodiments of another aspect of this application, the use of the ionizable lipid compound (ATILipid) targeting atherosclerotic cells described in the first aspect of this application is provided in the preparation of drug-loaded liposomes, drug-loaded lipid nanoparticles, drug-loaded polymer nanoparticles, drug-loaded nanomicelles, or nanoemulsion pharmaceutical formulations.

[0285] In some embodiments of another aspect of this application, a method for treating inflammation in vitro is provided, comprising administering to cells an ionizable lipid compound (ATILipid) as described in the first aspect of this application, a delivery carrier composition as described in the second aspect of this application, a drug delivery carrier (such as liposomes or lipid nanoparticles) as described in one aspect of this application, a bioactive composition, bioactive complex, bioactive formulation, pharmaceutical composition, drug-loaded complex, or pharmaceutical formulation as described in the third aspect of this application.

[0286] In some embodiments of another aspect of this application, a method of treating a disease is provided, comprising administering to a subject a therapeutically effective amount of the ionizable lipid compound (ATILipid) described in the first aspect of this application, the delivery carrier composition described in the second aspect of this application, the drug delivery carrier (such as liposomes or lipid nanoparticles) described in one aspect of this application, the bioactive composition, bioactive complex, bioactive formulation, pharmaceutical composition, drug-loaded complex, or pharmaceutical formulation described in the third aspect of this application. The definition of a disease can be found in the fourth aspect of this application.

[0287] In some embodiments of another aspect of this application, methods for preparing ionizable lipid compounds (ATILipid), delivery carrier compositions, drug delivery carriers, bioactive compositions, bioactive complexes, bioactive formulations, pharmaceutical compositions, pharmaceutical complexes, or pharmaceutical formulations are provided.

[0288] As used in this article, "therapeutic effective amount" refers to the amount required to elicit a biological or medical response in an individual, such as the amount of reagent that brings about a positive physiological and / or pharmacological effect on an individual, including but not limited to improving symptoms, alleviating symptoms, slowing or delaying the progression of disease, or preventing disease.

[0289] As used in this article, "subject" refers to an animal, which can be a mammal. The term "mammal" mainly refers to warm-blooded vertebrate mammals, including but not limited to: such as humans, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, rats, pigs, cattle, sheep, horses, etc.

[0290] In some implementations, the subjects are humans or mice.

[0291] In some of these implementations, the subject is a human.

[0292] There are no particular restrictions on dosage form and administration method. Representative administration methods include, but are not limited to, injection, oral administration, and inhalation.

[0293] In some embodiments of the fifth aspect of this application, a method for preparing a bioactive complex or bioactive preparation is provided, comprising the following steps:

[0294] S10: Dissolve each component of the delivery carrier composition described in the second aspect of this application in ethanol to prepare a lipid ethanol solution;

[0295] S30: Using any one of the following methods, namely microfluidic method, thin film hydration combined with electroporation method, and pressure homogenization combined with shearing treatment method, a lipid ethanol solution is mixed with an aqueous solution of bioactive ingredient to prepare a bioactive complex or a liquid bioactive preparation including a bioactive complex; wherein, the bioactive aqueous solution is an aqueous solution containing bioactive ingredient.

[0296] In some embodiments, the method for preparing a bioactive agent further includes the step of drying a liquid bioactive agent to prepare a solid bioactive agent.

[0297] In some other embodiments of the fifth aspect of this application, a method for preparing a drug-loaded complex or pharmaceutical formulation is provided, comprising the following steps:

[0298] S20: Dissolve each component of the delivery carrier composition described in the second aspect of this application in ethanol to prepare a lipid ethanol solution;

[0299] S40: Using any one of microfluidic method, thin film hydration combined with electroporation method, and pressure homogenization combined with shearing method, a lipid ethanol solution and a drug aqueous solution are mixed and compounded to prepare a drug-loaded complex or a liquid drug formulation including a drug-loaded complex; the drug formulation is as defined in the third aspect of this application, and the drug-loaded complex is the drug-loaded complex in the drug formulation described in the third aspect of this application; wherein, the drug aqueous solution is an aqueous solution containing a bioactive ingredient (such as a drug ingredient, further such as a drug active ingredient).

[0300] In some implementations, the bioactive ingredient is a nucleic acid-like substance.

[0301] In some embodiments, the bioactive complex (such as a drug-loaded complex) is a lipid nanoparticle, liposome, or lipid nanoemulsion.

[0302] In some embodiments, the method for preparing a pharmaceutical preparation further includes the step of drying a liquid pharmaceutical preparation to form a solid pharmaceutical preparation.

[0303] In some embodiments, the liquid formulation contains a dispersion liquid, which further includes water and may or may not include a buffer. Without limitation, the buffer may include one or more of a salt component of a phosphate buffer solution, trimethylolpropanesulfonic acid, sodium citrate, citric acid, sodium dihydrogen phosphate, potassium dihydrogen phosphate, and 4-hydroxyethylpiperazine ethanesulfonic acid.

[0304] In some embodiments, the mass percentage of the dispersion in the liquid bioactive formulation (such as a liquid pharmaceutical formulation) may be 95% to 99.9%, optionally 98% to 99.9%, or any of the following percentages or a range selected from any two of the following percentages: 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.9%, etc.

[0305] In some embodiments, the mass percentage of water in the liquid bioactive preparation (such as a liquid pharmaceutical preparation) may be 90% to 99%, optionally 95% to 99%, or any of the following percentages or a range selected from any two of the following percentages: 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, etc.

[0306] In another aspect of this application, a method for preparing an ionizable lipid compound (ATILipid) targeting atherosclerotic cells is provided, which can be used to prepare the compound ATILibid described in the first aspect of this application.

[0307] Those skilled in the art can select appropriate synthetic methods based on the structure of the chosen compound ATILipid, including but not limited to methods of coupling reaction, methods of protecting and deprotecting functional groups, etc.

[0308] In some embodiments, the preparation method of compound ATILipid includes the following steps:

[0309] S100: An N-branched molecule (denoted as compound M1) is coupled with an unsaturated fatty acid or its carboxyl activated form (denoted as compound M2) with anti-inflammatory activity to prepare an N-branched intermediate (denoted as compound M3) with two unsaturated fatty acyl groups.

[0310] Among them, G3-F 31 F is a precursor group of group G. 31 It is a reactive group or its protected form; after step S200, G3-F 31 The residues form the group G;

[0311] The definitions of Z1, Z2, L1 and L2 can be found in the context of this application, for example, in the definitions in formula (I);

[0312] R 12 The definition is consistent with R1 or R2;

[0313] S200: The compound M3 or its deprotected form is coupled with a molecule (which can be referred to as compound M4) that can target the function of atherosclerotic cells to prepare the compound shown in formula (IA);

[0314] F 32 F is a reactive group. 32 With F 31 Or its deprotected form constitutes a functional group pair capable of coupling reaction, after step S200, R 03 -F 32 The residues form the group R3;

[0315] G, R 03 The definitions of R and R3 can be found in the context of this application, for example, in the definition in formula (I).

[0316]

[0317] In step S200, F 32 With F 31Covalent linkages can be formed by coupling reactions of the deprotected form or other forms. Non-limiting examples of the formed covalent linkages include -NH-, -CH2NH-*, etc. *-C(=O)NH-, *-NHC(=O)-, -SS-, *-C(=O)S-, *-SC(=O)-, *-OC(=O)-, or *-C(=O)O-; where * is closer to R3 and the other end is closer to G. Exemplary examples of functional group pairs undergoing coupling reactions include maleimide and thiol (click reaction), aldehyde and amino (imine reduction reaction), carboxyl or its activated form with amino-NH2 (amidation reaction), carboxyl or its activated form with hydroxyl (esterification reaction), carboxyl or its activated form with thiol (thioesterification reaction), etc.

[0318] F 31 Non-limiting examples include protected primary amino groups (-NH2), such as tert-butoxycarbonyl (Boc-NH-), (Fmoc-NH-), etc., which can be deprotected and converted to primary amino groups before step S200; further, using R 03 Taking a polypeptide unit as an example, the primary amino group can be amidated with the carboxyl group at the C-terminus or side end of the polypeptide unit, or the carboxyl group can be activated and then coupled to form an amide bond.

[0319] Non-limiting examples of activated forms of the carboxyl group include active ester groups, such as succinimide carboxylic acid ester groups.

[0320] It is understandable that the G3-F of compound M3 31 Some of the compounds can be introduced along with the N-branched molecule, or two unsaturated fatty acyl groups can be first connected to the N-branching center to prepare a branched molecule (denoted as compound M5), and then compound 3 can be prepared by modification based on F1.

[0321]

[0322] In some embodiments, taking the compound shown in formula (V) as an example, the synthetic reaction can be as follows, including steps (a) to (g):

[0323]

[0324] The definitions of R1, R2 and R3 can be found in the context of this application, for example, in the definitions in equation (I).

[0325] In some embodiments, the compound shown in formula (V) is the same as the compound shown in formula (IV).

[0326] In some embodiments, R1 and R2 are derived from EPA; G comprises a divalent chain aliphatic hydrocarbon group containing at least one ionizable site; alternatively, R3 may be a VCAM-1 targeting peptide unit containing the VHPKQHRC peptide. In some embodiments, the number of ionizable sites may be one. The ionizable site may be provided by an ionizable nitrogen-containing group.

[0327] Those skilled in the art can use known methods in the field of chemical synthesis to identify the structures of the raw materials, intermediates, and products involved in the preparation of compound ATILipid. These methods include, but are not limited to, proton NMR spectroscopy. 1 Methods include 1H NMR, mass spectrometry (MS), Fourier transform infrared spectroscopy (FT-IR), gas chromatography-mass spectrometry (GC-MS), and circular dichroism (CD).

[0328] The embodiments of this application will be described in detail below with reference to some examples. It should be understood that these embodiments are only for illustrating this application and are not intended to limit the scope of this application. For experimental methods in the following embodiments where conditions are not specified, please refer to the guidelines given in this application first, or follow experimental manuals or conventional conditions in the art, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0329] In the following examples, the measurement parameters of the raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0330] In the following examples, unless otherwise specified, the room temperature is 20℃~30℃, such as 25℃.

[0331] I. Synthesis and Characterization of ATILipid, an Ionizable Lipid Compound with Anti-inflammatory Activity

[0332] Example 1.

[0333] The synthetic route for compound 1 is as follows:

[0334]

[0335] The synthesis steps of compound 1 are as follows:

[0336] (1) Synthesis of compound a: N-(3-aminopropyl)-diethanolamine (1.85 mmol), methanol (10 mL), chloroform (10 mL), triethylamine (2.78 mmol), and 4-dimethylaminopyridine (DMAP, 5.49 mmol) were added sequentially to a round-bottom flask, followed by the slow addition of di-tert-butyl dicarbonate (Boc2O, 2.78 mmol). The reaction was allowed to proceed overnight. The organic solvent was removed by rotary evaporation, and the product was purified by silica gel column chromatography.

[0337] (2) Synthesis of compound b: Boc-protected N-(3-aminopropyl)-diethanolamine (1.72 mmol), dichloromethane (20 mL), DMAP (4-dimethylaminopyridine, 5.15 mmol), and EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, 5.15 mmol) were added to a round-bottom flask, followed by eicosapentaenoic acid (5.15 mmol), and the mixture was stirred overnight. The reaction was completed the next day, the organic solvent was removed by rotary evaporation, and the product was purified by silica gel column chromatography.

[0338] (3) Synthesis of compound c: Compound b was dissolved in dichloromethane (10 mL), trifluoroacetic acid (2 mL) was added, the mixture was stirred for 2 h, the reaction was quenched with saturated sodium bicarbonate, the organic phase was extracted with ethyl acetate, and the product 3 was obtained by rotary evaporation.

[0339] (4) Synthesis of compound d: Compound c (0.137 mmol), N-Boc-3-bromopropylamine (Boc-NH-CH2CH2CH2-NH2, 0.151 mmol) and acetonitrile (10 mL) were added to a round-bottom flask and reacted at 70 °C for 7-8 h. The organic solvent was removed by rotary evaporation and the product was purified by silica gel column chromatography to obtain product d.

[0340] (5) Synthesis of compound e: Compound d (0.127 mmol) was dissolved in dichloromethane (10 mL), trifluoroacetic acid (2 mL) was added, the reaction was stirred for 2 h, the reaction was quenched with saturated sodium bicarbonate, the organic phase was extracted with ethyl acetate, and product e was obtained by rotary evaporation.

[0341] (6) Synthesis of compound f: Compound e (0.127 mmol) and GMBS (N-γ-maleimide butyryl-oxysuccinimide ester, 0.191 mmol) were dissolved in dichloromethane (10 mL), reacted at room temperature, stirred overnight, the organic solvent was removed by rotary evaporation, and the product was purified by silica gel column chromatography to obtain product f.

[0342] (7) Synthesis of Compound 1: Compound f (0.105 mmol) and the VCAM-1 targeting peptide with the sequence VHPKQHRC (0.07 mmol) were dissolved in DMSO (dimethyl sulfoxide) under nitrogen protection and reacted overnight at room temperature. The reaction solution was dialyzed in pure water for 24–48 h using a 1000 Da dialysis bag. The aqueous phase obtained by dialyzing was freeze-dried to obtain Compound 1.

[0343] The structure of the prepared ionizable lipid compound with anti-inflammatory activity was identified using the following method: the lipid material was weighed, dissolved in deuterated chloroform, and the synthesized material was characterized by proton NMR spectroscopy. 1 1H NMR spectrum; the lipid material was weighed, dissolved in methanol, and characterized by mass spectrometry. The reaction product was dissolved in deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (deuterated DMSO) and analyzed using an NMR spectrometer (AVANCE III 500). 1 H NMR testing, obtained 1 1H NMR spectrum. The reaction product was dissolved in methanol and characterized by mass spectrometry (instrument model: Finnigan LTQ) to obtain the mass spectrometry results.

[0344] Compound f: 1 H-NMR: δ (ppm): 0.88 (6H), 1.42 (3H), 1.50 (2H), 1.56 (6H), 1.60 (6H), 2.28 (14H); 2.50 (16H);2.82(4H);3.06(2H);4.04(2H);4.31(4H);5.53(20H);7.01(2H);8.32(1H)[M+H + ]:953.7.

[0345] Compound 1: 1 H-NMR: δ(ppm):0.88(6H),0.91(6H)1.42(5H),1.50(9H),2.1(35H);2.50(12H);2 .77(20H);2.82(10H);4.10(10H);5.53(20H);6.81(4H);7.33(4H)8.17(12H)[M+H + ]:1958.2.

[0346] Compound f and Compound 1 1 H-NMR spectrum as follows Figure 8 As shown, the mass spectrometry results for compound 1 are as follows: Figure 9As shown, the chemical shift δ5-6 ppm corresponds to the characteristic peak of hydrogen atoms in the unsaturated bonds of aliphatic carbon chains; the chemical shift δ2-3 ppm corresponds to the characteristic peak of hydrogen atoms in saturated bonds; and the chemical shift δ7-8 ppm corresponds to the characteristic peak of hydrogen atoms on the carbon atoms of maleimide double bonds. In compound 1, this peak disappears, and a characteristic peak on the carbon atom of the aromatic ring (imidazolium ring) in histidine (H) appears near the chemical shift δ8-9 ppm, indicating successful peptide coupling. The mass spectrum of compound 1 shows molecular ion peaks at 490.3, 653.4, 979.6, and 1958.2, corresponding to [M+4]+ / 4, [M+3]+ / 3, [M+2]+ / 2, and [M+1]+, respectively, indicating that compound 1 was successfully prepared.

[0347] Other ionizable lipid compounds with anti-inflammatory activity listed in Table 3 can be prepared using a method similar to that used to prepare compound 1, with the main difference being one or more of the following steps:

[0348] (1) Replace the fatty acid (eicosapentaenoic acid) used in the preparation of compound b with other types of unsaturated fatty acids with anti-inflammatory activity (e.g., compounds 2, 3, 4, 5, and 6);

[0349] (2) When introducing maleimide group based on compound c, the amine molecules used to introduce N-linked groups are different (for example, in compound 3, N-Boc-3-bromopropylamine is replaced with Boc-NH-CH2CH2-N(CH3)-CH2CH2CH2-NH2);

[0350] (3) Replace the VCAM-1 targeting peptide with the sequence VHPKQHRC used in the preparation of compound 1 with a different targeting molecule. The targeting molecule used is another type of targeting peptide that can target atherosclerotic cells (e.g., compound 4, compound 5) or (e.g., compound 6);

[0351] (44) The step of introducing a maleimide group is omitted, based on the coupling reaction of compound e with the corresponding target molecule (e.g., compound 5 and compound 6); it is understood that the target peptide used can be in the form of an amino protected group, which is removed from the N-terminal amino protecting group (e.g., Fmoc) after the coupling reaction to obtain the corresponding ionizable lipid compound.

[0352] All compounds in Table 3 can be obtained through... 1 H NMR and mass spectrometry were used to confirm the structures of intermediates and ionizable lipid compounds.

[0353] Table 3. Structures of ionizable lipid compounds with anti-inflammatory activity

[0354]

[0355]

[0356] The peptides involved in Table 3 show their amino acid sequences from the N-terminus to the C-terminus, and are linked to G in Formula I through a side thiol or carboxyl group at the C-terminus.

[0357] II. Preparation and Characterization of Drug Nanocarrier Complexes (Drug-Loaded Lipid Formulations)

[0358] (I) Preparation of drug-loaded lipid formulations

[0359] The composition formulations shown in Table 4 are used to prepare drug-loaded lipid formulations containing the aforementioned "ionizable lipid compounds with anti-inflammatory activity" to obtain drug-loaded complexes as LNP formulations. The amount of dispersion solution used is 0.5-3 mL, and in each formulation in Table 4, the mass percentage of the dispersion solution is 98%-99.9%, of which the mass percentage of water is 95%-99%.

[0360] Table 4. Composition of drug-loaded lipid formulations of ATILipid, an ionizable lipid compound with anti-inflammatory activity.

[0361]

[0362]

[0363] In Table 4, "Other lipid materials" is the sum of steroids, auxiliary lipids, and polyethylene glycol-modified lipids; when other lipid materials include polyethylene glycol-modified lipids, the weight-average molecular weight of the PEG portion is approximately 1500 Da to 2500 Da.

[0364] Table 4 lists the following types of "steroids": cholesterol, stigmasterol, lanosterol, β-sitosterol, β-amylostosterol, and brassicosterol;

[0365] The “auxiliary lipids” mentioned in Table 4 are as follows: DSPC represents distearylphosphatidylcholine; DSPE represents distearylphosphatidylethanolamine; DMPE represents dimyristoylphosphatidylethanolamine; DMPC represents dimyristoylphosphatidylcholine; DPPC represents dipalmitoylphosphatidylcholine; DPPE represents dipalmitoylphosphatidylethanolamine; egg yolk lecithin; soybean lecithin;

[0366] Table 4 lists the following “polyethylene glycol-modified lipids”: DSPE-PEG represents polyethylene glycol-distearate phosphatidylethanolamine; DMG-PEG represents polyethylene glycol-dimyristoyl glyceride.

[0367] The “loaded drugs” mentioned in Table 4 are as follows: Rapamycin represents rapamycin (CAS: 53123-88-9); Atorvastatin represents atorvastatin (CAS: 134523-03-8); miR-100 represents Hsa-miR-100-p (3'-AACCCGUAGAUCCGAACUUGUG-5', SEQ ID No. 8); miR-199 represents Hsa-miR-199-3p (3'-AUUGGUUACAGUCUGAUGACA-5', SEQ ID No. 9); miR-99 represents Has-miR-99a (3'-CAAGCUCGCUUCUAUGGGUCUG-5', SEQ ID No. 10); mTOR siRNA (5'-AAGAAUCAAAGAGCAGAGC-3', SEQ ID No. 11); MMP9 peptide (CTTHWGFTLC, SEQ ID No. 8). No. 12); Polypeptide SOD (HCDGGLAGLSC, SEQ ID No. 13).

[0368] Table 5.

[0369]

[0370] Example 2.

[0371] Preparation of Formulas 1-1 to 1-10: Weigh the specified amount of lipid material (ionizable lipid compounds, other lipid materials, and optional pharmaceutical excipients; taking Formula 1-1 as an example: ionizable lipid compounds (ionizable lipids), DSPC, cholesterol, and DSPE-PEG) and dissolve them in anhydrous ethanol. Weigh the specified amount of the loaded drug and dissolve it in an acidic (pH = 4.0-5.0) buffered saline solution. Prepare lipid nanoparticles using microfluidic technology. Take two 1mL syringes and draw 900μL of the aqueous phase and 300μL of the ethanol phase, respectively. Set the process parameters as follows: total volume 1.2mL, aqueous phase to ethanol phase flow rate ratio 3:1, total flow rate 9mL / min, discard 0.3mL of the initial solution and 0.05mL of the final solution. Collect the sample solution using a centrifuge tube. After sample collection, place the sample in a 1000Da dialysis bag and dialyze it with PBS buffer to obtain the final product.

[0372] Example 3.

[0373] Preparation of formulations 2-1 to 2-3: Weigh a specified amount of ionizable lipid compound (ionizable lipid) and other lipid materials and excipients, dissolve them in an organic solvent, and then place them in a rotary evaporator for rotary evaporation to form a lipid film on the bottle wall. Weigh a specified amount of the loaded drug and dissolve it in phosphate buffer, add it to the lipid film, rotate it in a 50°C water bath for 2 hours, then sonicate it in a water bath for 30 minutes, and finally filter it through a 0.22 μm filter membrane to obtain the final product.

[0374] Example 4.

[0375] Preparation of Formulas 3-1, 3-2 and 4-1: Weigh a specified amount of ionizable lipid compound (ionizable lipid), other lipid materials and optional other pharmaceutical excipients, dissolve them in oleic acid to form the oil phase, weigh a specified amount of loaded drug and dissolve it in ultrapure water to form the aqueous phase, mix the oil phase and aqueous phase, and use a high-speed shearing machine to obtain the pre-emulsion, place the pre-emulsion in a high-pressure homogenizer to prepare the final product.

[0376] Example 5.

[0377] Preparation of the control group formulation: Specified amounts of lipid materials (DLin-MC3-DMA, DSPC, cholesterol, and DSPE-PEG2000) were weighed and dissolved in anhydrous ethanol. The molar percentages of each component were as follows: Dlin-MC3-DMA 50%, cholesterol 38.5%, DSPC 10%, and DSPE-PEG2000 1.5%. Blank lipid nanoparticles MC3-LNP were prepared using microfluidic technology with ultrapure water as the aqueous phase, serving as the control group.

[0378] “Dlin-MC3-DMA”, CAS number 1224606-06-7, is a commercially available synthetic lipid with the chemical composition of methyl 4-(N,N-dimethylamino)butyrate (dilinoleyl) ester.

[0379] (II) Characterization of drug nanocarrier complexes

[0380] The particle size, potential, and pKa of the drug nanocarrier complex injection solution were tested using the following method.

[0381] Particle size testing method: Dilute the sample with deionized water, select an appropriate particle size cell according to the type of measurement, inject the sample and place it in the measuring instrument, measure the particle size of the sample at room temperature, measure each group three times, take the average value and record it as "particle size".

[0382] Potential testing method: Dilute the sample with deionized water, select an appropriate potential cell according to the type of measurement, inject the sample and place it in the measuring instrument, measure the potential of the sample at room temperature, measure each group three times, take the average value and record it as "potential".

[0383] The particle size and potential test results can be found in Table 6, and can also be found in [other documents / references]. Figure 1-3 .

[0384] Table 6. Particle size, potential and apparent Pka of the formulation

[0385] prescription Particle size (nm) Potential / (mV) Apparent pKa Prescription 1-1 69.04 8.695 5.92 Prescription 1-2 73.94 8.570 5.96 Prescription 1-3 74.45 9.638 6.07 Prescriptions 1-4 64.50 9.710 6.08 Prescriptions 1-5 76.56 8.070 5.94 Prescriptions 1-6 64.81 9.930 5.97 Prescriptions 1-7 75.01 9.600 6.01 Prescriptions 1-8 65.26 8.312 6.05 Prescriptions 1-9 62.34 8.890 6.03 Prescription 1-10 65.36 7.361 5.99 Prescription 2-1 79.13 9.405 / Prescription 2-2 72.55 8.530 / Prescription 2-3 73.10 9.806 / Prescription 3-1 93.64 7.105 / Prescription 3-2 91.79 8.560 / Prescription 4-1 92.05 8.686 /

[0386] pKa determination: For a 96-well plate system, prepare a series of buffer solutions with pH values ​​between 3 and 10. Dissolve the nanoparticles in the corresponding gradient solutions, add TNS (2-(p-toluidine)-6-naphthalenesulfonic acid), and place the plate in a multi-functional microplate reader for fluorescence reading (Ex / Em = 325nm / 435nm). The fluorescence intensity readings and pH values ​​are then curve-fitted to calculate the pKa.

[0387] The ionizable lipid compounds prepared according to the technical solution of this application have suitable pKa values ​​ranging from 5.5 to 6.5. Test results for some formulations can be found in [reference needed]. Figure 3 .

[0388] (III) Biological Effects Testing

[0389] 3.1 Cell-targeted uptake experiment

[0390] Human umbilical vein endothelial cells (HUVECs) were cultured in DMEM medium and seeded at an appropriate density in 12-well plates. Red fluorescent dye DiI was added as a marker during formulation preparation. When cells reached 60%-70% confluence, the cell culture medium was discarded, and DiI-loaded formulation diluted in blank medium was added and co-incubated with the cells for 6 hours. After incubation, the formulation was removed, cells were washed, digested with trypsin, and the supernatant was discarded by centrifugation. Cells were resuspended in PBS, and targeted uptake of the formulation by cells was detected by flow cytometry; alternatively, cells were fixed with 4% paraformaldehyde after incubation, and nuclear staining was performed with DAPI (4',6-diamidinyl-2-phenylindole, blue fluorescent dye). After slide preparation, the uptake of the formulation by cells was observed under a laser confocal microscope (Olymplus FV3000).

[0391] The results show that the drug nanocarrier complex prepared according to the technical solution of this application effectively promotes targeted uptake by cells. Test results for some prescription formulations can be found in [reference needed]. Figure 4 .

[0392] 3.2 Nucleic acid transfection level

[0393] HUVEC cells were spaced at 2 × 10⁶ cells per well. 5Cells were seeded in 6-well plates and cultured for 48 hours after the cells reached 80% cell growth. The LNP preparation was diluted with DMEM medium containing 1% embryonic bovine serum. Total protein was extracted after 48 hours, and Western blotting was used to detect the knockdown level of the target gene and the expression level of autophagy-related proteins.

[0394] The drug nanocarrier complex prepared according to the technical solution of this application effectively promotes nucleic acid transfection and successfully knocks down the mTOR protein. Test results for some prescription formulations are available for reference. Figure 5 .

[0395] 3.3 Investigation of anti-inflammatory activity

[0396] HUVEC cells were spaced at 1 × 10⁶ cells per well. 4 Cells were seeded in 96-well plates and, when the cells reached 80% cell growth, the LNP preparation was diluted with DMEM medium containing 1% embryonic bovine serum. Different lipid concentration gradients (corresponding to different incubation concentrations) were established based on total lipid mass: 0 mg / mL, 0.2 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.8 mg / mL, and 1.0 mg / mL. Cells were then incubated in a cell culture incubator for 48 h. After 48 h, the supernatant was collected, centrifuged, and the supernatant was retained. The concentration of IL-6 in the supernatant was detected by ELISA.

[0397] The drug nanocarrier complex prepared according to the technical solution of this application exhibits anti-inflammatory effects at different incubation concentrations, reducing the secretion of inflammatory factors by cells; the test results of the experimental group with an incubation concentration of 0.5 mg / mL are provided in the reference. Figure 6 It has a good anti-inflammatory effect and reduces the secretion of inflammatory factors by cells.

[0398] 3.4 Treatment efficacy for atherosclerosis

[0399] Animal experiments were conducted using 6-week-old male ApoE - / - C57 mice were first acclimatized for one week, then fed a mixture of high-fat diet (HF10014) and their regular diet, gradually increasing the proportion of high-fat diet until they were entirely fed a high-fat diet. After 12 weeks of this regimen, mice with large, obvious atherosclerotic plaques in their aorta were obtained and then administered medication. Every three days, 100 μL of a formulation containing an effective concentration of 10 mg / mL of an ionizable lipid compound was administered via tail vein injection. After seven administrations, the aorta of the mice was removed, and the area of ​​the plaques in the aorta was observed using Oil Red (Beyond) staining.

[0400] Test results are available for reference. Figure 7The drug nanocarrier complexes prepared according to the technical solutions of this application, such as formulations 1-1, 1-5, and 3-2, can effectively treat atherosclerosis and reduce plaque formation, among which formulations 1-1 and 1-5 are more effective.

[0401] III. Reference Proportion

[0402] Reference compound 1.

[0403]

[0404] The test was conducted using the method described in "(III) Bioeffects Test". The results showed that the single-tailed lipid was more toxic and the membrane disturbance caused by the single tailed chain was less effective. Compared with compound 1, the in vivo transfection effect and therapeutic effect of reference preparation 1 were significantly worse.

[0405] The technical features of the above embodiments and examples can be combined in any suitable manner. For the sake of brevity, not all possible combinations of the technical features in the above embodiments and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0406] The above embodiments and examples are merely illustrative of several implementations of this application, intended to facilitate a detailed understanding of the technical solutions of this application, but should not be construed as limiting the scope of protection of this application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the scope of protection of this application. Furthermore, it should be understood that after reading the above teachings, those skilled in the art can make various alterations or modifications to this application, and the equivalent forms obtained also fall within the scope of protection of this application. It should also be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided in this application through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this patent application should be determined by the content of the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. An ionizable lipid compound targeting atherosclerotic cells, characterized in that, The ionizable lipid compound is a compound with the structure shown in formula (I); In formula (I), R1- and R2- each independently form an unsaturated fatty acyl group with anti-inflammatory activity with the carbonyl group; Z1 and Z2 are each independently O, NH or S; L1 and L2 are each independently an alkylene group with 1 to 3 carbon atoms in the main chain; G is a divalent aliphatic hydrocarbon group containing one or more ionizable N atoms; R3 includes structural units R that can target the function of atherosclerotic cells. 03 .

2. The ionizable lipid compound according to claim 1, characterized in that, It meets one or more of the following characteristics: (a1) R1-C(=O)- and R2-C(=O)- each independently contain 18 to 22 carbon atoms; (a2) R1-C(=O)- and R2-C(=O)- each independently contain 1 to 6 carbon-carbon double bonds; (a3) R1- and R2- each independently form an unsaturated fatty acyl group derived from ω-3 polyunsaturated fatty acids with the carbonyl group; (a4) L1 and L2 are each independently -(CR a R b ) q -, where R a and R b Each is independently a H atom or a methyl group, and q is an integer selected from 1 to 3; Optionally, L1 and L2 are each independently C 1-3 Alkylene.

3. An ionizable lipid compound targeting atherosclerotic cells, characterized in that, The ionizable lipid compound is a compound with the structure shown in formula (I); In formula (I), R1- and R2- independently form an unsaturated fatty acyl group with the carbonyl group, derived from α-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, medelic acid, or sinapic acid. Z1 and Z2 are each independently O, NH or S; L1 and L2 are each independently C 1-3 Alkylene; G is a divalent aliphatic hydrocarbon group containing one or more ionizable N atoms; R3 includes structural units R that have the function of targeting atherosclerotic cells. 03 .

4. The ionizable lipid compound according to any one of claims 1 to 3, characterized in that, It meets one or more of the following characteristics: (b1)R 01 -and R 02 -Each group independently forms one of the following acyl groups with a carbonyl group: Alternatively, R1-C(=O)- and R2-C(=O)- are each independently a fatty acyl group derived from eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA); (b2) Z1 and Z2 are each independently O; (b3) L1 and L2 are each independently -(CH2) p -, where p is an integer selected from 1 to 3; Optionally, L1 and L2 are each independently -CH2CH2-; (b4)G includes a divalent group G0, wherein G0 is composed of one or more straight-chain saturated alkylene groups and one or more ionizable groups, wherein the ionizable groups are selected from at least one of ionizable -NH-, ionizable -N(CH3)- and a six-membered nitrogen heterocycle containing an ionizable nitrogen atom; Optionally, the six-membered nitrogen heterocycle containing an ionizable nitrogen atom is selected from at least one of piperidine ring and piperazine ring; Optionally, G0 is composed of one or more straight-chain saturated alkylene groups and one or more ionizable nitrogen-containing groups; more preferably, G0 is composed of one or two straight-chain saturated alkylene groups and one ionizable nitrogen-containing group. Optionally, the straight-chain saturated alkylene group is C10. 2-3 Straight-chain saturated alkylene groups; Optionally, G0 is selected from at least one of the following divalent groups: Alternatively, G0 is (b5)R 03 It includes at least one of the functional residues of the following substances: small molecule organic compounds and polypeptides or proteins having at least 4 amino acid units; (b6)R 03 Includes at least one of the functional residues of the following substances: polypeptides, monoclonal antibodies, and carbohydrate molecules; Optionally, R 03 Including polypeptide structural units and At least one of them; Further optional, R 03 Including targeted peptide units based on one or more of the following polypeptides: VCAM-1 targeted peptide, ICAM-1 targeted peptide, P-selectin targeted peptide, S2P targeted peptide, IL-4 receptor targeted peptide, MCP-1 targeted peptide, CCR2 targeted peptide, collagen targeted peptide, CD44 targeted peptide, CD36 targeted peptide, SR-B1 targeted peptide, LOX-1 targeted peptide, fibrin targeted peptide, matrix metalloproteinase targeted peptide, integrin αvβ3 targeted peptide, oxidized low-density lipoprotein targeted peptide, and inflammasome targeted peptide; Furthermore, alternatively, R 03 Includes VCAM-1 targeting peptide units; Furthermore, alternatively, R 03 This includes VCAM-1 targeting peptide units containing VHPK peptides, or R3 is any of the following polypeptide structural units: VHPKQHRC, cyclic CNNSKSHTC (cyclic), VHPKQHRGGSGC, VHPKQHRGGSKGC, cyclic FLDRK (cyclic), and cyclic NNSKSHT (cyclic); wherein the (cyclic) in the cyclic polypeptide structural unit indicates that the amino acid units at both ends are linked by peptide bonds to form a ring; (b7) G and R3 are linked by at least one of the following structures: chemical bond, -NH-, -CH2NH-*, *-C(=O)NH-, *-NHC(=O)-, -SS-, *-C(=O)S-, *-SC(=O)-, *-OC(=O)- or *-C(=O)O-; where * is closer to R3 and the other end is closer to G; (b8) R3-G- is selected from any of the following groups: (b9) The ionizable lipid compound described herein has the structure shown in formula (III):

5. The ionizable lipid compound according to any one of claims 1 to 3, characterized in that, The ionizable lipid compound is any of the following compounds:

6. A delivery carrier composition, characterized in that, The delivery carrier composition, by mass percentage, comprises 20%–70% of an ionizable lipid compound, 0%–60% of a steroid, 0%–30% of an auxiliary lipid, 0%–10% of a polyethylene glycol-modified lipid, and 0%–15% of other pharmaceutical excipients; or, by mass percentage, the delivery carrier composition comprises 20%–90% of an ionizable lipid compound, 0%–50% of a steroid, 0%–50% of an auxiliary lipid, 0%–20% of a polyethylene glycol-modified lipid, and 0%–80% of other pharmaceutical excipients. Wherein, the ionizable lipid compound is at least one of the ionizable lipid compounds targeting atherosclerotic cells as described in any one of claims 1 to 5; The polyethylene glycol-modified lipid is different from any of the ionizable lipid compounds, the steroids, and the auxiliary lipids; The steroid is different from either the auxiliary lipid or the PEGylated lipid; Optionally, the delivery carrier composition comprises at least one of the steroid, the auxiliary lipid, and the polyethylene glycol-modified lipid; Optionally, the auxiliary lipids include one or more of phosphatidylethanolamines, acylphosphatidylcholines, betaine lipids (DGTS), and sphingomyelins (SM); Optionally, the delivery carrier composition satisfies one or more of the following characteristics: (c1) The steroids include one or more of the following substances: cholesterol, lanosterol, sitosterol, stigmasterol, ergosterol, bastaminophen, campesterol, and campesterol; optionally, the steroids include cholesterol; (c2) The auxiliary lipids include at least one of distearylphosphatidylcholine (DSPC), distearylphosphatidylethanolamine (DSPE), dioleoylphosphatidylethanolamine (DOPE), dioleoyllecithin (DOPC), dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylethanolamine (DPPE), myristoylphosphatidylcholine (DMPC), myristoylphosphatidylethanolamine (DMPE), 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine (POPC), dioleoylphosphatidylglycerol (DOPG), betaine lipids (DGTS), and sphingomyelin (SM); optionally, the auxiliary lipids include one or more of DSPC, DSPE, DOPE, and DOPC; (c3) The lipids in the polyethylene glycol-modified lipids include one or more of the following: dimyristoyl glycerol (DMG), distearyl phosphatidylethanolamine (DSPE), distearyl glycerol (DSG), dipalmitoyl glycerol, dioleoyl glycerol, distearyl glycerol, diacylglycamide (DAG), dipalmitoyl phosphatidylethanolamine (DPPE), polyethyleneimine (PE), succinyl-diacylglycerol (S-DAG), dialkoxypropyl carbamate, and dimethacrylate (c-DMA); (c4) The weight-average molecular weight of the polyethylene glycol portion in the polyethylene glycol-modified lipid is 1500 Da to 2500 Da; (c5) The polyethylene glycol portion of the polyethylene glycol-modified lipid includes polyethylene glycol monomethyl ether, polyethylene glycol hydroxyl or polyethylene glycol amino; (c6) The PEGylated lipids include methoxy polyethylene glycol bis(tetradecyl acetamide) (ALC-0159), PEG-DMG (polyethylene glycol-dimyristoyl glycerol), PEG-DSPE (polyethylene glycol-distearate phosphatidylethanolamine), PEG-DSG (polyethylene glycol-distearate glycerol), PEG-DPG (polyethylene glycol-dipalmitoyl glycerol), PEG-DOPE (polyethylene glycol-dioleoyl phosphatidylethanolamine), PEG-distearate, and PEG-DAG (polyethylene glycol-diacylglycamide). The PEG-DPPE (polyethylene glycol-dipalmitoylphosphatidylethanolamine), PEG-PE (polyethylene glycol-polyethyleneimine), PEG-S-DAG (polyethylene glycol-succinoyl-diacylglycerol), PEG-dialkoxypropylcarbamate, and PEG-c-DMA (polyethylene glycol-dimethacrylate) may be selected from one or more of these. Optionally, the PEGylated lipids may include one or more of DMG-PEG2000, DSPE-MPEG2000, ALC-0159, and PEG-DOPE. (c7) The delivery carrier composition further includes other pharmaceutical excipients selected from one or more of the following pharmaceutical excipients: surfactants, emulsifiers, co-emulsifiers, solubilizers, buffers, osmotic pressure regulators, antibacterial agents and protein stabilizers; (c8) By mass percentage, the delivery carrier composition comprises 30%–70% of an ionizable lipid compound, 5%–60% of a steroid, 0%–30% of an auxiliary lipid, 0%–10% of a polyethylene glycol-modified lipid, and 0%–15% of other pharmaceutical excipients; or, by mass percentage, the delivery carrier composition comprises 40%–90% of an ionizable lipid compound, 5%–50% of a steroid, 0%–30% of an auxiliary lipid, 0%–10% of a polyethylene glycol-modified lipid, and 0%–80% of other pharmaceutical excipients; Optionally, by molar percentage, the delivery carrier composition comprises 30%–60% of an ionizable lipid compound, 15%–45% of a steroid, 5%–20% of an auxiliary lipid, 1%–5% of a polyethylene glycol-modified lipid, and 0%–15% of other pharmaceutical excipients. (c9) The molar percentage of the ionizable lipid compound in the delivery carrier composition is 30% to 70%, optionally 30% to 60%, or the mass percentage of the ionizable lipid compound in the delivery carrier composition is 30% to 90%. (c10) The sum of the mass percentages of the steroid, the auxiliary lipid, and the polyethylene glycol-modified lipid in the delivery carrier composition is 20% to 80%, optionally 30% to 80%; (c11) The steroid in the delivery carrier composition is 5% to 60% by molar percentage, optionally 15% to 45% by mass, or the steroid in the delivery carrier composition is 5% to 45% by mass. (c12) The auxiliary lipid in the delivery carrier composition has a molar percentage of 5% to 30%, optionally 5% to 20%, or the auxiliary lipid in the delivery carrier composition has a mass percentage of 5% to 50%. (c13) The polyethylene glycol-modified lipid in the delivery carrier composition is 1% to 5% by molar percentage, or the polyethylene glycol-modified lipid in the delivery carrier composition is 2% to 20% by mass. (c14) The delivery carrier composition further includes other pharmaceutical excipients, wherein the other pharmaceutical excipients are present in the delivery carrier composition at a molar percentage of 0-15%, optionally 5%-10%, or at a mass percentage of 10%-60%; (c15) The delivery carrier composition constitutes a drug delivery carrier, wherein the drug delivery carrier is a liposome, lipid nanoparticle, polymer nanoparticle, nanomicelle or nanoemulsion drug carrier. (c16) The average particle size of the drug delivery carrier is 50 nm to 200 nm, and can be selected as 50 nm to 100 nm.

7. A pharmaceutical composition, drug-loaded complex, or pharmaceutical formulation, characterized in that, The pharmaceutical formulation includes the drug-loaded complex; the pharmaceutical composition or the drug-loaded complex independently includes a bioactive ingredient and an ionizable lipid compound targeting atherosclerotic cells as described in any one of claims 1 to 5.

8. The pharmaceutical composition, drug-loaded complex, or pharmaceutical formulation according to claim 7, characterized in that, It meets one or more of the following characteristics: (d1) The pharmaceutical composition or the drug-loaded complex further comprises independently at least one of steroids, auxiliary lipids, and PEGylated lipids; wherein the auxiliary lipids comprise one or more of phosphatidylethanolamines, phosphatidylcholine derivatives, betaine lipids (DGTS), and sphingomyelin (SM); (d2) The average particle size of the drug-loaded complex is 50 nm to 200 nm, and can be selected as 50 nm to 100 nm; (d3) The drug-loaded complex is a drug-loaded liposome, drug-loaded lipid nanoparticle, drug-loaded polymer nanoparticle, drug-loaded nanomicelle or nanoemulsion drug formulation. (d4) The bioactive ingredients include at least one of nucleic acid substances, proteins, polypeptides and small molecule organic drugs; (d5) The bioactive ingredient is negatively charged or is a hydrophobic substance; (d6) The bioactive components include nucleic acid substances, which include one or more of miRNA, mRNA, siRNA, ASO, saRNA, shRNA, Pre-RNA, LncRNA, RNAmimics (RNA mimics), RNAagomir, RNA antagomir, DNA, and cDNA; (d7) The mass ratio of the bioactive component to the ionizable lipid compound is 1:(0.5 to 1000), and optionally 1:(1 to 200); (d8) The bioactive component includes nucleic acid substances, wherein the mass ratio of the nucleic acid substances to the ionizable lipid compound is 1:(1-1000), or optionally 1:(1-200); (d9) The pharmaceutical composition or the pharmaceutical formulation independently comprises the delivery carrier composition of claim 6; the bioactive ingredient is a nucleic acid substance; by mass percentage, the delivery carrier composition comprises 50%–70% of an ionizable lipid compound, 25%–35% of a steroid, 5%–30% of an auxiliary lipid, 1%–5% of a polyethylene glycol-modified lipid, and 5%–10% of other pharmaceutical excipients; the mass ratio of the nucleic acid substance to the ionizable lipid compound is 1:(10–100); (d10) The pharmaceutical formulation is a liquid formulation, and the solvent includes water; optionally, the mass percentage of the drug-loaded complex in the liquid formulation is 1% to 20%.

9. The ionizable lipid compound targeting atherosclerotic cells as described in any one of claims 1 to 5 is used in the preparation of a drug delivery carrier or drug-loaded complex, or in the preparation of a medicament for treating a disease; or, the ionizable lipid compound targeting atherosclerotic cells as described in any one of claims 1 to 5, the delivery carrier composition as described in claim 6, or the drug composition or drug-loaded complex as described in claim 7 or 8 is used in the preparation of a pharmaceutical formulation for treating a disease or in the preparation of a medicament for treating a disease. in, The diseases to be treated include at least one of the following: treatment of chronic inflammatory diseases, targeted therapy for inflammatory diseases, targeted therapy for atherosclerosis, treatment of atherosclerotic inflammation, treatment of familial hyperlipidemia, treatment of atherosclerotic lipid deposition, treatment of hypertension caused by atherosclerosis, treatment of atherosclerotic stroke, and treatment of atherosclerotic vascular damage; the inflammatory diseases include plaque inflammation; the chronic inflammatory diseases include cardiovascular diseases; Optionally, the disease includes at least one of immune-related inflammatory cardiovascular disease, atherosclerotic inflammation, atherosclerotic myocardial infarction, and atherosclerotic stroke.

10. A method for preparing a drug-loaded complex or pharmaceutical formulation, characterized in that, Includes the following steps: The components of the delivery carrier composition according to claim 6 are dissolved in ethanol to prepare a lipid ethanol solution; Using any one of microfluidic methods, thin-film hydration combined with electroporation, and pressurized homogenization combined with shearing, the lipid ethanol solution is mixed with a drug aqueous solution to prepare a drug-loaded complex or a liquid pharmaceutical formulation including a drug-loaded complex; the pharmaceutical formulation is as defined in claim 7 or 8, and the drug-loaded complex is the drug-loaded complex in the pharmaceutical formulation of claim 7 or 8; wherein, the drug aqueous solution is an aqueous solution containing bioactive components; Optionally, the bioactive component is a nucleic acid-like substance; Optionally, the drug-loaded complex is a lipid nanoparticle, a liposome, or a lipid nanoemulsion; Optionally, the method for preparing the pharmaceutical formulation further includes the step of drying the liquid pharmaceutical formulation to form a solid pharmaceutical formulation.

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