Complex for nucleic acid delivery

Abstract

The present disclosure provides nucleic acid particles comprising an immunomodulatory agent, an RNA, and a cationic lipid or cationic polymer, and the nucleic acid particles described herein reduce the inflammatory response associated with previous formulations and / or increase protein or antigen expression.

Description

Technical Field

[0001] Cross - reference to related applications This application claims priority to PCT Application No. PCT / EP22 / 64542, filed on May 30, 2022, the entire content of which is incorporated herein by reference.

Background Art

[0002] Nucleic acid particles are complexes useful for delivering nucleic acid therapies to patients. Generally, nucleic acid particles contain lipids (e.g., lipid nanoparticles (LNPs), liposomes, and lipoplexes), and / or cationic polymers (e.g., polyplexes), and a nucleic acid, e.g., ribonucleic acid (RNA).

Summary of the Invention

[0003] Nucleic acid therapies face certain challenges when administered to patients. In order to be effective, nucleic acid molecules need to reach the target tissue and produce a specific protein of interest. However, nucleic acids can be unstable, are susceptible to degradation after administration, and alone have limited ability to enter the target tissue. Therefore, there is a need for nucleic acid particle delivery technologies that encapsulate nucleic acids and facilitate delivery to the patient and the target of interest within the patient. In particular, nucleic acid particles containing lipids (e.g., lipid nanoparticles (LNPs), liposomes, and lipoplexes), as well as those containing polymers (e.g., polyplexes), have emerged as vehicles that facilitate the delivery of nucleic acids to the target of interest.

[0004] Particles containing cationic lipids and / or cationic polymers are carefully constructed vehicles that can rely on the interaction of different components to provide a safe means of transferring nucleic acid cargo. Lipid nanoparticles (LNPs), for example, contain cationic lipids and specific helper compounds that stabilize the particles to ensure that the LNP can release the cargo at the appropriate time and place. Certain particles, such as those containing lipids including LNP, are widely considered to be safe and effective for the delivery of nucleic acid therapies and are actually used in commercially available products worldwide, but still have the potential to cause an inflammatory response by the immune system after administration. For example, intradermal and intramuscular administration of LNP has been noted to induce "inflammation characterized by leukocyte infiltration, activation of different inflammatory pathways, and secretion of a diverse pool of inflammatory cytokines and chemokines." Ndeupen, et al., iScience, 24, 103479 (Dec. 17, 2021). These responses can be characterized as pain, swelling, fever, etc.

[0005] This disclosure encompasses the insight that encapsulation of certain agents as part of nucleic acid particles, such as lipid-based particles (e.g., LNPs, lipoplexes, and liposomes), as well as polymer-based particles (e.g., polyplexes), can reduce the inflammatory response upon administration and, in some embodiments, further increase translation. Without wishing to be bound by theory, it is hypothesized that the provided nucleic acid particles not only retain stability for targeted delivery of the nucleic acid, but also reduce the inflammatory response in the patient, thereby avoiding certain side effects such as pain and fever. The nucleic acid particles described herein incorporate these agents into the structure of the particle itself. Furthermore, the nucleic acid particles described herein surprisingly also show improved expression of the protein or antigen encoded by the nucleic acid delivered by the particle.

[0006] Other attempts to reduce such side effects have focused on modifying specific components of the nanoparticles, including, for example, the modification of cationic lipids to incorporate specific steroid-like structural features. In contrast, the present disclosure directly incorporates immunomodulatory compounds, such as TLR inhibitors and / or inflammasome inhibitors, into the structure of the particles as separate agents.

[0007] Furthermore, attempts have also been made to use dexamethasone to reduce inflammation upon administration of formulations containing nucleic acids (e.g., RNA). See, for example, Chen, et al., J. of Controlled Release, 286:46-54 (2018), Zhang, et al., J. Biomed. Mater. Res. 2022;1-8; I. Vlatkovic, Biomedicines, 9:520 (2021)). The applicant has discovered alternative agents to solve the problem of the inflammatory response upon administration of nucleic acid particles.

[0008] In some embodiments, the present disclosure provides nucleic acid particles comprising RNA, an immunomodulatory agent, and a cationic lipid or cationic polymer. In some embodiments, the immunomodulatory agent is not dexamethasone.

[0009] In some embodiments, the nucleic acid particles comprise an immunomodulatory agent, a cationic lipid, and RNA. In some embodiments, the nucleic acid particles are in the form of lipid nanoparticles. In some embodiments, the lipid nanoparticles further comprise one or more of a helper lipid and a polymer-conjugated lipid.

[0010] In some embodiments, the nucleic acid particles comprise an immunomodulatory agent, a cationic polymer, and RNA.

[0011] In some embodiments, the present disclosure provides a method of increasing or causing an increase in the expression of RNA in a target of a subject, the method comprising administering to the subject the nucleic acid particles described herein.

[0012] In some embodiments, the present disclosure provides a method of treating a disease, disorder, or condition in a subject, the method comprising administering to the subject a nucleic acid particle described herein.

[0013] In some embodiments, the present disclosure provides a nucleic acid particle described herein for use as a medicament.

[0014] In some embodiments, the present disclosure provides a nucleic acid particle described herein for use in the treatment and / or prevention of a disease or disorder, wherein the disease or disorder is an infectious disease, cancer, genetic disorder, autoimmune disease, or rare disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0015]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Mode for Carrying Out the Invention

[0016] The present disclosure provides, inter alia, nucleic acid particles (e.g., cationic lipid - based particles such as lipid nanoparticles (LNPs), liposomes, lipoplexes, and cationic polymer - based particles such as polyplexes) useful for the delivery of nucleic acids, for example, RNA, and their use. The present disclosure also provides, inter alia, complexes that avoid problems associated with previously known nucleic acid particles, including an inflammatory response, or an increase in the levels of cytokines or interleukins that can cause pain, fever, and other adverse reactions after administration. As described herein, the present disclosure provides nucleic acid particles (and compositions, e.g., pharmaceutical compositions comprising such nucleic acid particles) comprising RNA, an immunomodulator, and a cationic lipid or cationic polymer. In some embodiments, the nucleic acid particles described herein are useful for the treatment of various diseases. In some embodiments, such nucleic acid particles can be administered via systemic, intravenous, or intranasal means.

[0017] Definition The compounds of the present disclosure generally include those described previously and are further exemplified by the classes, subclasses, and species disclosed herein. As used herein, unless otherwise indicated, the following definitions shall apply. For the purposes of the present disclosure, chemical elements are identified by the Periodic Table, CAS version, Handbook of Chemistry and Physics, 75th Edition. Further, the general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, as well as "March’s Advanced Organic Chemistry", 5th Edition: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

[0018] Unless otherwise indicated, the structures shown herein are intended to include all stereoisomeric forms (e.g., enantiomers or diastereomers) of the structure, as well as all geometric isomeric forms or conformational isomeric forms of the structure. For example, the R and S configurations of each stereocenter are contemplated as part of the present disclosure. Therefore, single stereochemical isomers of the provided compounds, as well as enantiomers, diastereomers, and geometric (or conformational) mixtures are within the scope of the present disclosure. Unless otherwise specified, all tautomeric forms of the provided compounds are within the scope of the present disclosure.

[0019] Unless otherwise specified, the structures shown herein are meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, substitution of hydrogen by deuterium or tritium, or 13 C or 14 substitution of carbon by

[0020] About or approximately: As used herein, the terms "about" or "approximately" when applied to one or more target values refer to a value similar to the recited reference value. Generally, one of ordinary skill in the art familiar with the context will fully understand the degree of relevant difference subsumed by "about" or "approximately" in that context. For example, in some embodiments, the term "about" or "approximately" may encompass a range of values within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the reference value (i.e., ±).

[0021] Administration: As used herein, the terms "administering" or "administration" typically refer to the administration of a composition (e.g., a pharmaceutical composition) to a subject to achieve delivery of the composition or an agent contained therein to a target site or site to be treated. One of ordinary skill in the art will recognize the various routes available for administration to a subject, such as a human, under appropriate circumstances. For example, in some embodiments, administration can be ocular, oral, parenteral, topical, etc. In some particular embodiments, administration can be bronchial (e.g., by bronchial instillation), buccal, percutaneous (e.g., one or more of or including topical skin, intradermal, interdermal, transdermal, etc.), enteral, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, into a particular organ (e.g., into the liver), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by tracheal instillation), vaginal, vitreous, etc. In some embodiments, administration can be parenteral. In some embodiments, administration can be oral. In some particular embodiments, administration can be intravenous. In some particular embodiments, administration can be subcutaneous. In some embodiments, administration can involve only a single dose. In some embodiments, administration can involve the application of a defined number of doses. In some embodiments, administration can involve intermittent (e.g., multiple doses separated in time) and / or periodic (e.g., individual doses separated by a common period) administration. In some embodiments, administration can involve continuous administration (e.g., perfusion) for at least a selected period. In some embodiments, administration can include a prime and boost protocol. A prime and boost protocol can include the administration of a first dose of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine), followed by the administration of a second or subsequent dose of the pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) after a defined time interval. In the case of an immunogenic composition, a prime and boost protocol can result in an increased immune response in a patient.

[0022] Agonist: As used herein, the term "agonist" generally refers to an agent whose presence or level correlates with an increase in the level or activity of a target, compared to what is observed in the absence of the agent (or compared to different levels of the agent). In some embodiments, an agonist is an agent whose presence or level correlates with a target level or activity that is comparable to, or greater than, a particular reference level or activity (e.g., what is observed under appropriate reference conditions such as the presence of a known agonist, e.g., a positive control). In some embodiments, an agonist may be a direct agonist in that it directly affects (e.g., directly interacts with) the target, and in some embodiments, an agonist may be an indirect agonist in that it affects the target indirectly (e.g., acts on a regulator of the target, or some other component or entity, e.g., by interacting with it).

[0023] Aliphatic: The term "aliphatic" means a straight-chain (i.e., unbranched), branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon that is completely saturated or contains one or more unsaturated units but is not aromatic (also referred to herein as "cycloaliphatic") and has a single or multiple point of attachment to the remainder of the molecule. Unless otherwise specified, aliphatic groups contain from 1 to 12 aliphatic carbon atoms. In some embodiments, an aliphatic group contains from 1 to 6 aliphatic carbon atoms (e.g., C 1-6 ). In some embodiments, an aliphatic group contains from 1 to 5 aliphatic carbon atoms (e.g., C 1-5 ). In other embodiments, an aliphatic group contains from 1 to 4 aliphatic carbon atoms (e.g., C 1-4 ). In still other embodiments, an aliphatic group contains from 1 to 3 aliphatic carbon atoms (e.g., C 1-3 ), and in still other embodiments, an aliphatic group contains from 1 to 2 aliphatic carbon atoms (e.g., C 1-2) is included. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl groups, and hybrids thereof. Preferred aliphatic groups are C 1-6 alkyl.

[0024] Alkyl: The term "alkyl", used alone or as part of a larger moiety, means a saturated, optionally substituted, straight or branched chain hydrocarbon group having from 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, 1 to 3, or 1 to 2 carbon atoms (unless otherwise specified) (e.g., C 1-12 , C 1-10 , C 1-8 , C 1-6 , C 1-4 , C 1-3 , or C 1-2 ). Exemplary alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, and heptyl.

[0025] Alkylene: The term "alkylene" refers to a divalent alkyl group. In some embodiments, "alkylene" is a divalent straight or branched alkyl group. In some embodiments, an "alkylene chain" is a polymethylene group, i.e., -(CH 2 ) n-where n is a positive integer, for example, 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. The optionally substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are optionally substituted with substituents. Suitable substituents include those described below for substituted aliphatic groups and also those described in the specification of this application. It will be understood that two substituents of the alkylene group can together form a ring system. In certain embodiments, two substituents can together form a 3- to 7-membered ring. The substituents can be on the same or different atoms. When added to a particular group in this application, the suffix “-ene” or “-enyl” is intended to refer to the bifunctional portion of the group. For example, when “-ene” or “-enyl” is added to “cyclopropyl”, it becomes “cyclopropylene” or “cyclopropylenyl”, a bifunctional cyclopropyl group, for example, [Chemical Formula] is intended to refer to.

[0026] Alkenyl: The term “alkenyl”, used alone or as part of a larger moiety, has at least one double bond and has 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, or 2 to 3 carbon atoms (unless otherwise specified) (e.g., C 2-12 , C 2-10 , C 2-8 , C 2-6 , C 2-4 , or C 2-3 ) and means an optionally substituted straight-chain or branched-chain or cyclic hydrocarbon group. Exemplary alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, and heptenyl. The term “cycloalkenyl” means an optionally substituted non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond and having about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, and cycloheptenyl.

[0027] Alkynyl: The term "alkynyl", used alone or as part of a larger moiety, has at least one triple bond and has 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, or 2 to 3 carbon atoms (unless otherwise specified) (e.g., C 2-12 , C 2-10 , C 2-8 , C 2-6 , C 2-4 , or C 2-3 ), and means a straight-chain or branched-chain hydrocarbon group, optionally substituted. Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and heptynyl.

[0028] Antagonist: As understood by those skilled in the art, the term "antagonist" generally refers to an agent whose presence or level correlates with a decrease in the level or activity of a target as compared to that observed in the absence of the agent (or compared to different levels of the agent). In some embodiments, the antagonist is one whose presence or level correlates with a target level or activity that is comparable to or less than a particular reference level or activity (e.g., that observed under appropriate reference conditions such as the presence of a known antagonist, e.g., a positive control). In some embodiments, the antagonist may be a direct antagonist in that it directly affects (e.g., directly interacts with) the target, and in some embodiments, the antagonist may be an indirect antagonist in that it affects the target indirectly (e.g., by acting on a regulator of the target, or some other component or entity, e.g., by interacting with it).

[0029] Aryl: The term "aryl" has a total of 6 to 14 ring members (e.g., C 6 -C 14 ), and means a monocyclic or bicyclic ring system in which at least one ring within the system is aromatic and each ring within the system contains 3 to 7 ring members. In some embodiments, the "aryl" group has a total of 6 to 12 ring members (e.g., C6 -C 12 ) and includes. The term "aryl" may be used synonymously with the term "aryl ring". In certain embodiments, "aryl" refers to an aromatic ring system including phenyl, biphenyl, naphthyl, anthracyl, etc., and may have one or more substituents. Unless otherwise specified, the "aryl" group is a hydrocarbon. In some embodiments, the "aryl" ring system is an aromatic ring (e.g., phenyl) condensed with a non-aromatic ring (e.g., cycloalkyl). Examples of the condensed aryl ring include

Chemical formula

[0030] Biological sample: As used herein, the term "biological sample" typically means a sample obtained from, or derived from, a biological source (e.g., tissue or organism or cell culture) for the purposes described herein. In some embodiments, the source of interest includes organisms such as animals or humans. In some embodiments, the biological sample is or includes a biological tissue or fluid. In some embodiments, the biological sample can be or include bone marrow, blood, blood cells, ascites, tissue or fine needle biopsy sample, cell-containing body fluid, cell-free nucleic acid, sputum, saliva, urine, cerebrospinal fluid, peritoneal fluid, pleural effusion, feces, lymph, gynecological fluid, skin swab, vaginal swab, oral swab, nasal swab, lavage or wash fluid such as ductal lavage or bronchoalveolar lavage, aspirate, scrape, bone marrow specimen, tissue biopsy specimen, surgical specimen, feces, other body fluids, secretions, and / or excretions, and / or cells therefrom, etc. In some embodiments, the biological sample is or includes cells obtained from an individual. In some embodiments, the obtained cells are or include cells from the individual from whom the sample was obtained. In some embodiments, the sample is a "primary sample" obtained directly from the source of interest by any suitable means. For example, in some embodiments, the primary biological sample is obtained by a method selected from the group consisting of biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of body fluid (e.g., blood, lymph, feces, etc.). In some embodiments, as will be apparent from the context, the term "sample" means a preparation obtained by processing the primary sample (e.g., by removing one or more components and / or adding one or more agents). For example, filtration using a semipermeable membrane. Such a "processed sample" can include, for example, nucleic acids or proteins obtained by subjecting the primary sample to techniques such as extraction from the sample, or amplification or reverse transcription of mRNA, isolation and / or purification of specific components.

[0031] Carrier: As used herein, the term "carrier" means a diluent, adjuvant, excipient, or vehicle used in administering a composition. In some exemplary embodiments, the carrier can include, for example, water and a sterile liquid such as an oil including, for example, peanut oil, soybean oil, mineral oil, sesame oil, or other oils of petroleum, animal, vegetable, or synthetic origin. In some embodiments, the carrier is or includes one or more solid components.

[0032] Combination therapy: As used herein, the term "combination therapy" refers to a situation where a subject is simultaneously exposed to two or more treatment regimens (e.g., two or more therapeutic agents or modalities). In some embodiments, the two or more regimens may be administered simultaneously. In some embodiments, these regimens may be administered sequentially (e.g., all "doses" of the first regimen are administered before any dose of the second regimen). In some embodiments, these agents are administered in an overlapping dosing regimen. In some embodiments, "administration" of combination therapy can include administering one or more agents or modalities in combination to a subject that is receiving another agent(s) or modality(ies). For clarity, combination therapy does not require that the individual agents be administered together (or even necessarily simultaneously) in a single composition, although in some embodiments, two or more agents or their active moieties may be administered together in a combined composition or even a combined compound (e.g., as part of a single chemical complex or covalent conjugate).

[0033] Comparable: As used herein, the term "comparable" refers to two or more sets of agents, substances, situations, conditions, etc., which need not be identical to each other, but are sufficiently similar to enable a comparison between them such that a conclusion can be reasonably drawn based on the observed differences or similarities, as understood by one of ordinary skill in the art. In some embodiments, comparable sets of conditions, situations, individuals, or groups are characterized by a plurality of substantially identical features and one or a few varying characteristics. One of ordinary skill in the art will understand, in any given context, the degree of identity required for two or more such sets of agents, substances, situations, conditions, etc. to be considered comparable. For example, one of ordinary skill in the art will understand that a series of situations, individuals, or groups are comparable to each other if the differences in the results or observed phenomena obtained under or using different series of situations, individuals, or groups are caused by variations in their varying features or are characterized by a sufficient number and variety of substantially identical features to warrant a reasonable conclusion indicating variations in the features.

[0034] Composition: One of ordinary skill in the art will understand that the term "composition" can be used to refer to a distinct physical entity containing one or more defined components. Generally, unless otherwise specified, a composition can be in any form, such as a gas, gel, liquid, solid, etc.

[0035] Cycloaliphatic: As used herein, the term "cycloaliphatic" refers to a monocyclic C 3-8 hydrocarbon or bicyclic C 6-10 hydrocarbon that is either fully saturated or contains one or more units of unsaturation, is not aromatic, and has a single-point or multiple-point attachment to the remainder of the molecule.

[0036] Cycloalkyl: As used herein, the term "cycloalkyl" refers to an optionally substituted saturated monocyclic or polycyclic ring system of about 3 to about 10 ring carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0037] Dosage form or unit dosage form: One of ordinary skill in the art will understand that the term "dosage form" can be used to refer to physically discrete units of an active agent (e.g., a therapeutic or diagnostic agent) for administration to a subject. Typically, each such unit contains a predetermined amount of the active agent. In some embodiments, such amount is an appropriate unit dose (or a whole thereof) for administration according to a dosing regimen (i.e., a therapeutic dosing regimen) that has been determined to correlate with a desired or beneficial result when administered to a relevant population.

[0038] Dosage Regimen or Treatment Regimen: One of ordinary skill in the art will understand that the terms "dosage regimen" and "treatment regimen" can typically be used to refer to a set of unit doses (typically more than one) that are administered to a subject individually over time. In some embodiments, a given therapeutic agent may have a recommended dosage regimen that includes one or more doses. In some embodiments, the dosage regimen includes multiple doses where each dose is temporally spaced from other doses. In some embodiments, the individual doses are separated from each other at equal intervals. In some embodiments, the dosage regimen includes multiple doses and individual doses separated in two different periods. In some embodiments, all the doses within a dosage regimen are of the same unit dose. In some embodiments, the different doses within a dosage regimen are of different amounts. In some embodiments, the dosage regimen includes a first administration at a first dosage and one or more additional administrations at a second dosage that is different from the first dosage. In some embodiments, the dosage regimen includes a first administration at a first dosage and one or more additional administrations at a second dosage that is the same as the first dosage. In some embodiments, the dosage regimen is correlated with a desired or beneficial outcome when administered within a relevant population (i.e., is a therapeutic dosage regimen).

[0039] Excipient: As used herein, the term "excipient" means a non-therapeutic agent that can be included in a pharmaceutical composition, for example, to provide or contribute to a desired viscosity or stabilizing effect. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, and the like.

[0040] Heteroaliphatic: As used herein, the term "heteroaliphatic" or "heteroaliphatic group" can be linear (i.e., unbranched), branched, or cyclic ("heterocyclic"), can be fully saturated, or can contain one or more unsaturated units, but is not aromatic, and refers to an optionally substituted hydrocarbon moiety having 1 to 5 heteroatoms in addition to carbon atoms. The term "heteroatom" means nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of basic nitrogen. The term "nitrogen" also includes substituted nitrogen. Unless otherwise specified, a heteroaliphatic group contains 1 to 10 carbon atoms, and 1 to 3 carbon atoms are optionally and independently substituted with heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, the heteroaliphatic group contains 1 to 4 carbon atoms, and 1 to 2 carbon atoms are optionally and independently substituted with heteroatoms selected from oxygen, nitrogen, and sulfur. In still other embodiments, the heteroaliphatic group contains 1 to 3 carbon atoms, and 1 carbon atom is optionally and independently substituted with a heteroatom selected from oxygen, nitrogen, and sulfur. Suitable heteroaliphatic groups include, but are not limited to, linear or branched heteroalkyl groups, heteroalkenyl groups, and heteroalkynyl groups. For example, heteroaliphatic groups of 1 to 10 atoms include exemplary groups such as -O-CH 3 -, -CH 2 -O-CH 3 -, -O-CH 2 -CH 2 -O-CH 2 -CH 2 -O-CH 3 and the like.

[0041] Heteroaryl: The terms "heteroaryl" and "heteroaral-" are used alone or as part of a larger moiety, for example, "heteroaralkyl" or "heteroaralkoxy" have 5 to 10 ring atoms, have 6, 10, or 14 π electrons shared in a cyclic array, and have 1 to 5 heteroatoms in addition to carbon atoms, and refer to monocyclic or bicyclic ring groups (e.g., 5- to 6-membered monocyclic heteroaryl or 9- to 10-membered bicyclic heteroaryl). Examples of heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl, imidazo[1,2-a]pyrimidinyl, imidazo[1,2-a]pyridyl, imidazo[4,5-b]pyridyl, imidazo[4,5-c]pyridyl, pyrrolopyridyl, pyrrolopyrazinyl, thienopyrimidinyl, triazolopyridyl, and benzisoxazolyl. As used herein, the terms "heteroaryl" and "heteroar-" include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings and the radical or point of attachment is on the heteroaromatic ring (i.e., a bicyclic heteroaryl ring having 1 to 3 heteroatoms). Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzotriazolyl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyrido[2,3-b]-1,4-oxazin-3(4H)-one, 4H-thieno[3,2-b]pyrrole, and benzisoxazolyl.The term "heteroaryl" can be used interchangeably with the terms "heteroaryl ring", "heteroaryl group", or "heteroaromatic", and any of these terms includes a ring that is optionally substituted.

[0042] Heteroatom: As used herein, the term "heteroatom" means nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur and any quaternized form of basic nitrogen.

[0043] Heterocyclic ring: As used herein, the terms "heterocyclic ring", "heterocyclyl", "heterocyclic radical", and "heterocyclic ring" are used interchangeably and are either saturated or partially unsaturated and refer to a stable 3- to 8-membered monocyclic, 6- to 10-membered bicyclic, or 10- to 16-membered polycyclic heterocyclic moiety having, in addition to carbon atoms, one or more, preferably 1 to 4, of the heteroatoms defined above. When used with respect to the ring atoms of a heterocyclic ring, the term "nitrogen" includes substituted nitrogen. By way of example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur, or nitrogen, nitrogen is N (such as in 3,4-dihydro-2H-pyrrolyl), NH (such as in pyrrolidinyl), or NR +can be (such as N-substituted pyrrolidinyl). The heterocyclic ring may be attached to its pendant group by any heteroatom or carbon atom so as to provide a stable structure, and any of the ring atoms may optionally be substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and thiomorpholinyl. The heterocyclyl group can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. The bicyclic heterocyclic ring also includes groups in which the heterocyclic ring is fused to one or more aryl rings. Exemplary bicyclic heterocyclic groups include indolinyl, isoindolinyl, benzodioxolyl, 1,3-dihydroisobenzofuranyl, 2,3-dihydrobenzofuranyl, and tetrahydroquinolinyl. The bicyclic heterocyclic ring can be a spirocyclic ring system (e.g., a 7- to 11-membered spirocyclic fused heterocyclic ring having, in addition to carbon atoms, one or more heteroatoms as defined above (e.g., 1, 2, 3, or 4 heteroatoms)). The bicyclic heterocyclic ring can also be a bridged ring system (e.g., a 7- to 11-membered bridged heterocyclic ring having 1, 2, or 3 bridging atoms).

[0044] Lipid: As used herein, the term "lipid" refers to a molecule that also includes one or more hydrophobic moieties or groups, and optionally one or more hydrophilic moieties or groups. Molecules containing both hydrophobic and hydrophilic moieties are also frequently referred to as amphiphilic substances. Lipids are usually insoluble in water. In an aqueous environment, due to their amphiphilic nature, molecules can self-organize into organized structures and different phases. Since they are present in vesicles, multilamellar / unilamellar liposomes, or membranes in an aqueous environment, one of these phases consists of a lipid bilayer. Hydrophobicity can be conferred by including nonpolar groups including, but not limited to, long-chain saturated and unsaturated aliphatic hydrocarbon groups, and such groups substituted by one or more aromatic groups, alicyclic groups, or heterocyclic groups. In some embodiments, the hydrophilic group can include polar and / or charged groups and can include carbohydrates, phosphates, carboxylic acids, sulfates, aminos, sulfhydryls, nitros, hydroxyls, and other similar groups.

[0045] Modulator: As used herein, the term "modulator" refers to a compound (e.g., a small molecule) that can change the activity of another molecule (e.g., a protein). For example, in some embodiments, a modulator can cause an increase or decrease in the magnitude of the activity of a particular target molecule as compared to the magnitude of its activity in the absence of the modulator. For example, a modulator can be an agonist or an antagonist of a particular target as those terms are defined herein. For example, in some embodiments, the modulator is an agonist. In some embodiments, the modulator is an antagonist.

[0046] Oral: As used herein, the phrases "oral administration" and "administered orally" have the meaning understood in the art and mean oral administration of a compound or composition.

[0047] Parenteral: As used herein, the terms "parenteral administration" and "administered parenterally" have the meaning understood in the art and generally mean a mode of administration other than enteral and topical administration by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subdural, intraspinal, and intrasternal injection and infusion.

[0048] Partially unsaturated: As used herein, the term "partially unsaturated" refers to a ring moiety that contains at least one double or triple bond between ring atoms. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic (e.g., aryl or heteroaryl) moieties as defined herein.

[0049] Patient or subject: As used herein, the terms "patient" or "subject" mean any organism to which the provided composition is or can be administered for, e.g., experimental, diagnostic, prophylactic, cosmetic, and / or therapeutic purposes. Typical patients or subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, the patient is human. In some embodiments, the patient or subject has or is susceptible to one or more disorders or conditions. In some embodiments, the patient or subject exhibits one or more symptoms of a disorder or condition. In some embodiments, the patient or subject has been diagnosed with one or more disorders or conditions. In some embodiments, the patient or subject has received or has previously received a particular therapy for diagnosing and / or treating a disease, disorder, or condition.

[0050] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dosage appropriate for administration in a treatment regimen or dosing regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition is formulated for administration in solid or liquid form, including oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., as a sterile solution or suspension, or as a sustained release formulation, by, e.g., subcutaneous, intramuscular, intravenous, or epidural injection; topical application to the skin, lung, or oral cavity, e.g., as a cream, ointment, or controlled release patch or spray; intravaginal or rectal, e.g., as a pessary, cream, or foam; sublingual; ocular; transdermal; or suitable for application to the nose, lung, and other mucosal surfaces.

[0051] Pharmaceutically acceptable: As used herein, the phrase "pharmaceutically acceptable" means a compound, material, composition, and / or dosage form that, within the scope of sound medical judgment, is suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications and that has a reasonable benefit / risk ratio.

[0052] Pharmaceutically acceptable salt: As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound suitable for use in a pharmaceutical context, i.e., a salt that, within the scope of sound medical judgment, is suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, etc. and that has a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S.M. Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66:1-19 (1977).

[0053] Polypeptide: As used herein, the term "polypeptide" typically has the meaning recognized in the art of a polymer of at least three or more amino acids. Those skilled in the art will understand that the term "polypeptide" encompasses not only polypeptides having the complete sequences described herein, but also polypeptides representing functional, biologically active, or characteristic fragments, portions, or domains of such complete polypeptides (e.g., fragments, portions, or domains that retain at least one activity). In some embodiments, the polypeptide may contain L-amino acids, D-amino acids, or both, and / or may contain any of various amino acid modifications or analogs known in the art. Useful modifications include, for example, terminal acetylation, amidation, methylation, and the like. In some embodiments, the polypeptide may contain natural amino acids, non-natural amino acids, synthetic amino acids, and combinations thereof (e.g., may be or contain peptidomimetics).

[0054] Polymer: As used herein, the term "polymer" refers to a composition comprising one or more molecules containing repeating units of one or more monomers. As used herein, the terms "polymer" and "polymer composition" are used interchangeably and, unless otherwise specified, refer to a composition of polymer molecules. Those skilled in the art will understand that a polymer composition contains polymer molecules having different lengths (e.g., containing varying amounts of monomers). The polymer compositions described herein have a number average molecular weight (M n ), a weight average molecular weight (M w) and / or one or more of a polydispersity index (PDI). The polymers described herein may also be characterized by a degree of polymerization (DP) that refers to the number of monomer units in the polymer. The polymers described herein can be homopolymers, heteropolymers, or block copolymers. As used herein, "homopolymer" refers to a polymer having a single type of monomer that is repeated throughout the polymer chain, e.g., -A-A-A-A-. As used herein, "heteropolymer" refers to a polymer having more than one type (e.g., two or more) of monomers present throughout the polymer chain, e.g., -A-B-A-B-A-. As used herein, "block-copolymer" refers to a polymer having an arrangement of blocks of polymerized monomers, e.g., -A-A-A-A-B-B-B-B- (diblock polymer) or -A-A-A-A-B-B-B-B-A-A-A- (triblock polymer). The polymers described herein can be linear or branched.

[0055] Prevention or prophylaxis: As used herein, the terms "prevent" or "prophylaxis" when used in connection with the occurrence of a disease, disorder, and / or condition mean a reduction in the risk that the disease, disorder, and / or condition will occur, and / or a delay in the occurrence of one or more features or symptoms of the disease, disorder, or condition. Prophylaxis can be considered complete when the occurrence of the disease, disorder, or condition is delayed over a predetermined period of time.

[0056] Ribonucleotide: As used herein, the term "ribonucleotide" encompasses unmodified ribonucleotides and modified ribonucleotides. For example, unmodified ribonucleotides include the purine bases adenine (A) and guanine (G), and the pyrimidine bases cytosine (C) and uracil (U). Modified ribonucleotides can include, for example, (a) terminal modifications, such as 5'-terminal modifications (e.g., phosphorylation, dephosphorylation, conjugation, inverted linkage, etc.), 3'-terminal modifications (e.g., conjugation, inverted linkage, etc.), (b) base modifications, such as modified bases, stabilized bases, destabilized bases, or bases having an expanded repertoire of partners for base pairing or substitution with conjugate bases, (c) sugar modifications (e.g., at the 2'- or 4'-position) or sugar substitutions, and (d) internucleoside bond modifications, including modifications or substitutions of the phosphodiester bond, and can include one or more modifications, including but not limited to these. The term "ribonucleotide" also encompasses ribonucleotide triphosphates, including modified and unmodified ribonucleotide triphosphates.

[0057] Ribonucleic acid (RNA): As used herein, the term "RNA" refers to a polymer of ribonucleotides. In some embodiments, the RNA is single-stranded. In some embodiments, the RNA is double-stranded. In some embodiments, the RNA contains both single-stranded and double-stranded portions. In some embodiments, the RNA can include the backbone structure described in the above definition of "nucleic acid / polynucleotide". The RNA can be a regulatory RNA (e.g., siRNA, microRNA, etc.) or messenger RNA (mRNA). In some embodiments, the RNA is mRNA. In some embodiments, the RNA is mRNA, and the RNA typically includes a poly(A) region at its 3' end. In some embodiments, the RNA is mRNA, and the RNA typically includes a cap structure recognized in the art at its 5' end for, e.g., recognizing the mRNA to initiate translation and binding to ribosomes. In some embodiments, the RNA is synthetic RNA. Synthetic RNA includes RNA synthesized in vitro (e.g., by enzymatic synthesis and / or chemical synthesis methods). As used herein, "monomeric RNA", also referred to as "single-molecule RNA", refers to individual RNA molecules that are not aggregates, dimers, trimers, or oligomers of RNA.

[0058] Sample: As used herein, the term "sample" typically refers to an aliquot of material obtained from or derived from a source of interest. In some embodiments, the source of interest is a biological or environmental source. In some embodiments, the source of interest can be or can include cells, tissues, or organisms such as microorganisms, plants, or animals (e.g., humans). In some embodiments, the source of interest is or includes a biological tissue or fluid. In some embodiments, the source of interest can be or can include a preparation produced in a production process. In some embodiments, the sample is a "primary sample" obtained directly from the source of interest by any suitable means. In some embodiments, as will be apparent from the context, the term "sample" means a preparation obtained by processing the primary sample (e.g., by removing one or more components and / or adding one or more agents).

[0059] Substituted or optionally substituted: As described herein, the compounds of the present invention can include a "optionally substituted" moiety. In general, the term "substituted", whether or not preceded by the term "optionally", means that one or more hydrogens of the designated moiety are replaced by a suitable substituent. "Substituted" applies to one or more hydrogens that are either explicitly or implicitly part of the structure (e.g.,

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0060] Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are, independently, halogen, -(CH 2 ) 0-4 R°, -(CH 2 ) 0-4 OR°, -O(CH 2 ) 0-4 R°, -O-(CH 2 ) 0-4 C(O)OR°, -(CH 2 ) 0-4 CH(OR°) 2 、-(CH 2 ) 0-4 SR°, -(CH 2 ) 0-4 Ph that can be substituted with R°, -(CH 2 ) 0-4 O(CH 2 ) 0-1 Ph that can be substituted with R°, -CH=CHPh that can be substituted with R°, -(CH 2 ) 0-4 O(CH 2 ) 0-1 -pyridyl, -NO 2 、-CN、-N 3 、(CH2 ) 0-4 N(R°) 2 、-(CH 2 ) 0-4 N(R°)C(O)R°, -N(R°)C(S)R°, -(CH 2 ) 0-4 N(R°)C(O)NR° 2 、N(R°)C(S)NR° 2 、-(CH 2 ) 0-4 N(R°)C(O)OR°, -N(R°)N(R°)C(O)R°, N(R°)N(R°)C(O)NR° 2 、N(R°)N(R°)C(O)OR°, -(CH 2 ) 0-4 C(O)R°, C(S)R°, -(CH 2 ) 0-4 C(O)OR°, -(CH 2 ) 0-4 C(O)SR°, (CH 2 ) 0-4 C(O)OSiR° 3 、-(CH 2 ) 0-4 OC(O)R°, -OC(O)(CH 2 ) 0-4 SR°, -(CH 2 ) 0-4 SC(O)R°, -(CH 2 ) 0-4 C(O)NR° 2 、-C(S)NR° 2 、-C(S)SR°, -SC(S)SR°, (CH 2 ) 0-4 OC(O)NR° 2 、C(O)N(OR°)R°, -C(O)C(O)R°, -C(O)CH 2 C(O)R°, -C(NOR°)R°, (CH 2 ) 0-4 SSR°, -(CH 2 ) 0-4 S(O) 2 R°, -(CH 2 ) 0-4 S(O) 2 OR°, -(CH 2 ) 0-4 OS(O) 2 R°, -S(O)2 NR° 2 、(CH 2 ) 0-4 S(O)R°, N(R°)S(O) 2 NR° 2 、-N(R°)S(O) 2 R°, -N(OR°)R°, -C(NH)NR° 2 、-P(O) 2 R°, P(O)R° 2 、OP(O)R° 2 、-OP(O)(OR°) 2 、SiR° 3 、-(C 1-4 linear or branched alkylene)O-N(R°) 2 、or -(C 1-4 linear or branched alkylene)C(O)O-N(R°) 2 wherein each R° may be substituted as defined below and is independently hydrogen, C1-6 aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1 Ph, -CH 2 -(5- to 6-membered heteroaryl ring), or independently a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms selected from nitrogen, oxygen, or sulfur, or regardless of the above definition, the occurrence of two independent R°'s, together with the intervening atom(s), independently forms a 3- to 12-membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0 to 4 heteroatoms selected from nitrogen, oxygen, or sulfur, and these may be substituted as defined below.

[0061] Preferred monovalent substituents on R° (or the ring formed by the occurrence of two independent R°'s together with the intervening atoms) are independently halogen, -(CH 2 ) 0-2 R ● 、-(haloR ● )、-(CH 2 ) 0-2 OH, -(CH 2 ) 0-2 OR ● 、-(CH 2 ) 0-2CH(OR ● ) 2 、O(haloR ● )、 -CN、 -N 3 、 -(CH 2 ) 0-2 C(O)R ● 、 -(CH 2 ) 0-2 C(O)OH、 -(CH 2 ) 0-2 C(O)OR ● 、 -(CH 2 ) 0-2 SR ● 、 -(CH 2 ) 0-2 SH、 -(CH 2 ) 0-2 NH 2 、 -(CH 2 ) 0-2 NHR ● 、 -(CH 2 ) 0-2 NR ● 2 、 -NO 2 、 -SiR ● 3 、 -OSiR ● 3 、 C(O)SR ● 、 -(C 1-4 linear or branched alkylene)C(O)OR ● 、 or -SSR ● and each R ● is unsubstituted or, when preceded by "halo", substituted by only one or more halogens and independently is selected from C 1-4 aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1 Ph, or independently is selected from 3 - to 6 - membered saturated, partially unsaturated, or aryl rings having 0 to 4 heteroatoms selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on the saturated carbon atoms of the "optionally substituted" groups include =O and =S.

[0062] Suitable divalent substituents on the saturated carbon atoms of the "optionally substituted" groups include =O ("oxo"), =S, =NNR * 2 、 =NNHC(O)R *, =NNHC(O)OR * , =NNHS(O) 2 R * , =NR * , =NOR * , -O(C(R * 2 )) 2-3 , O- or -S(C(R * 2 )) 2-3 S- is exemplified, and each independent occurrence of R * is hydrogen, C that may be substituted as defined below 1-6 aliphatic, or independently selected from unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl rings having 0 to 4 heteroatoms selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that bond to an adjacent substitutable carbon of an "optionally substituted" group include -O(CR * 2 ) 2-3 O- is exemplified, and each independent occurrence of R * is hydrogen, C that may be substituted as defined below 1-6 aliphatic, or independently selected from unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl rings having 0 to 4 heteroatoms selected from nitrogen, oxygen, or sulfur.

[0063] R * Suitable substituents on the aliphatic group of are halogen, -R ● , (haloR ● ), OH, -OR ● , -O(haloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH 2 , -NHR ● , -NR ● 2 , or -NO 2 is exemplified, and each R ● is unsubstituted or, when "halo" precedes, is substituted only with one or more halogens and independently is C 1-4 aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1Ph, or independently, a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms selected from nitrogen, oxygen, or sulfur.

[0064] Suitable substituents on a nitrogen of a "optionally substituted" group include -R † , -NR † 2 , -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CH 2 C(O)R † , S(O) 2 R † , S(O) 2 NR † 2 , -C(S)NR † 2 , -C(NH)NR † 2 , or -N(R † )S(O) 2 R † where each R † is independently hydrogen, a C 1-6 aliphatic that may be substituted as defined below, unsubstituted -OPh, or independently, an unsubstituted 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms selected from nitrogen, oxygen, or sulfur, or regardless of the above definition, two independent occurrences of R † together with the intervening atom(s) therebetween independently form an unsubstituted 3- to 12-membered saturated, partially unsaturated, or monocyclic or bicyclic ring having 0 to 4 heteroatoms selected from nitrogen, oxygen, or sulfur.

[0065] R † Suitable substituents on the aliphatic group of are independently halogen, -R ● , (haloR ● ), -OH, -OR ● , -O(haloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH 2 , -NHR ● , -NR● 2 or NO 2 and each R ● is unsubstituted or, when "halo" precedes, is substituted with only one or more halogens and, independently, is C 1-4 aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1 Ph or, independently, is a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms selected from nitrogen, oxygen, or sulfur.

[0066] Small molecule: As used herein, the term "small molecule" means an organic compound and / or an inorganic compound of low molecular weight. Generally, a "small molecule" is a molecule having a size of less than about 5 kilodaltons (kD). In some embodiments, the small molecule is less than about 4 kD, 3 kD, about 2 kD, or about 1 kD. In some embodiments, the small molecule is less than about 800 daltons (D), about 600 D, about 500 D, about 400 D, about 300 D, about 200 D, or about 100 D. In some embodiments, the small molecule is less than about 2000 g / mol, less than about 1500 g / mol, less than about 1000 g / mol, less than about 800 g / mol, or less than about 500 g / mol. In some embodiments, the small molecule is not a polymer.

[0067] In some embodiments, the small molecule does not contain a polymer moiety. In some embodiments, the small molecule is not a protein or polypeptide and / or does not contain it (e.g., is not an oligopeptide or peptide). In some embodiments, the small molecule is not a polynucleotide and / or does not contain it (e.g., is not an oligonucleotide). In some embodiments, the small molecule is not a polysaccharide and / or does not contain it. For example, in some embodiments, the small molecule is not a glycoprotein, proteoglycan, glycolipid, etc. In some embodiments, the small molecule is not a lipid.

[0068] In some embodiments, the small molecule is a modulator (e.g., an inhibitor or activator). In some embodiments, the small molecule is biologically active. In some embodiments, the small molecule is detectable (e.g., includes at least one detectable moiety). In some embodiments, the small molecule is a therapeutic agent.

[0069] One of ordinary skill in the art, upon reading the present disclosure, will understand that the specific small molecule compounds described herein can be provided and / or utilized in any of a variety of forms, such as, for example, crystalline forms (e.g., polymorphs, solvates, etc.), salt forms, protected forms, prodrug forms, ester forms, isomeric forms (e.g., optical and / or structural isomers), isotopic forms, and the like.

[0070] One of ordinary skill in the art will understand that a particular small molecule compound can have a structure that can exist in one or more stereoisomeric forms. In some embodiments, such small molecules can be utilized in the form of individual enantiomers, diastereomers, or geometric isomers, or as a mixture of stereoisomers, according to the present disclosure, and in some embodiments, such small molecules can be utilized in the form of a racemic mixture, according to the present disclosure.

[0071] One of ordinary skill in the art will understand that a particular small molecule compound can have a structure that can exist in one or more tautomeric forms. In some embodiments, such small molecules can be utilized in the form of individual tautomers or in forms that interconvert between tautomers, according to the present disclosure.

[0072] One of ordinary skill in the art will understand that a particular small molecule compound can have a structure that allows for isotope substitution (e.g., in the case of H, 2 H or 3 H, 12 in the case of C, 11 C, 13 C, or 14 C, 14 in the case of N, 13 N or 15 N, 16 in the case of O, 17 O or 18 O,35 Cl or 37 In the case of Cl 36 Cl, 19 In the case of F 18 F, 127 In the case of I 131 I, etc.). It will be understood that some embodiments may utilize such small molecules in one or more isotopically modified forms, or mixtures thereof, in accordance with the present disclosure.

[0073] In some embodiments, reference to a particular small molecule compound may relate to a particular form of that compound. In some embodiments, a particular small molecule compound may be provided and / or utilized in salt form (e.g., in the form of an acid addition or base addition salt, depending on the compound), and in some such embodiments, the salt form may be in the form of a pharmaceutically acceptable salt.

[0074] In some embodiments, if a small molecule compound is a naturally occurring or naturally found compound, that compound may be provided and / or utilized in a form different from that in which it occurs naturally or is naturally found. One of ordinary skill in the art will understand that in some embodiments, the absolute or relative amounts of compounds or forms present in a reference preparation of interest (e.g., a primary sample from a source of interest such as a biological or environmental source) may be different (e.g., with respect to another component of a preparation containing a different form of the compound), and that a preparation containing an absolute or relative amount of a compound, or a particular form of a particular small molecule compound, that is different from the compounds present in the reference preparation or source. Thus, in some embodiments, for example, a preparation of a single stereoisomer of a small molecule compound may be considered a different form from a racemic mixture of the compound, a particular salt of a small molecule compound may be considered a different form from another salt form of the compound, a preparation containing only the form of a compound containing one stereoisomer ((Z) or (E)) of a double bond may be considered a different form from the form of a compound containing the other stereoisomer ((E) or (Z)) of the double bond, and a preparation in which one or more atoms are different isotopes from those present in the reference preparation may be considered a different form.

[0075] One of ordinary skill in the art will further understand that, in the context of a low molecular structure, as used herein, the symbol

Chem.

Chem.

[0076] Therapeutic agent: As used herein, the term "therapeutic agent" generally means any agent that, when administered to a living being, induces a desired pharmacological effect. In some embodiments, an agent is considered a therapeutic agent if it shows a statistically significant effect in an appropriate population. In some embodiments, the appropriate population can be a population of model organisms. In some embodiments, the appropriate population can be defined by various criteria such as a particular age group, sex, genetic background, existing clinical condition, etc. In some embodiments, a therapeutic agent is a substance that can be used to alleviate, ameliorate, reduce, inhibit, prevent, delay the onset of, decrease the severity of, and / or decrease the incidence of one or more symptoms or characteristics of a disease, disorder, and / or medical condition. In some embodiments, a "therapeutic agent" is an agent that has been approved by, or is required to be approved by, a government agency before it can be marketed for administration to humans. In some embodiments, a "therapeutic agent" is an agent that requires a medical prescription for administration to humans.

[0077] Treat: As used herein, the terms “treat,” “treatment,” or “treating” mean any method used to partially or completely alleviate, ameliorate, reduce, suppress, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or features of a disease, disorder, and / or condition. Treatment may be administered to a subject that does not exhibit symptoms of a disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject that exhibits only the initial symptoms of a disease, disorder, and / or condition, for the purpose of, for example, reducing the risk of developing a condition associated with the disease, disorder, and / or condition.

[0078] Complex for nucleic acid delivery The present disclosure provides nucleic acid particles (including compositions comprising such nucleic acid particles), which comprise a nucleic acid (e.g., RNA), an immunomodulatory agent, and a cationic lipid or cationic polymer. The present disclosure includes, inter alia, the surprising insight that certain agents, such as immunomodulatory agents, can be added to the nucleic acid particles described herein, causing a reduction in the inflammatory response seen with previous particles. Further, the nucleic acid particles described herein surprisingly also exhibit improved expression of a protein or antigen encoded by a nucleic acid delivered by the particles.

[0079] A "nucleic acid particle" (e.g., a ribonucleic acid particle) refers to a particle that contains or comprises a nucleic acid and is used to deliver the nucleic acid to a target site of interest (e.g., a cell, tissue, organ, etc.) as part of a composition (e.g., a pharmaceutical composition) that includes a plurality of nucleic acid particles. As described herein, a nucleic acid particle (e.g., a ribonucleic acid particle) can be formed from i) at least one cationic lipid or cationic ionizable lipid or lipid-like material, ii) at least one cationic polymer, such as polyethyleneimine, protamine, or a mixture thereof (i.e., a mixture of i) and ii)), and iii) a nucleic acid. Nucleic acid particles (e.g., ribonucleic acid particles) include lipid nanoparticles (LNPs), lipoplexes, liposomes, and polyplexes. In some embodiments, the nucleic acid particles described herein do not contain oligosaccharides.

[0080] In some embodiments, the particles for nucleic acid delivery described herein are lipid nanoparticles that include i) a nucleic acid, (ii) at least one cationic lipid or cationic ionizable lipid, and (iii) an immunomodulatory agent. In some embodiments, the particles for nucleic acid delivery include (i) a nucleic acid (e.g., DNA or RNA, e.g., mRNA), (ii) at least one cationic lipid or cationic ionizable lipid disclosed herein, (iii) an immunomodulatory agent, and (iv) at least one additional lipid described herein (e.g., a steroid, a helper lipid (also referred to as a "neutral lipid"), a polymer-conjugated lipid, or a combination thereof).

[0081] Electrostatic interactions between positively charged molecules such as cationic polymers and cationic lipids and negatively charged nucleic acids are involved in particle formation. This results in complexation and spontaneous formation of nucleic acid particles.

[0082] Although not intended to be bound by any theory, it is believed that cationic lipids or cationic ionizable lipids or lipid-like materials, and / or cationic polymers complex with nucleic acids to form aggregates, and colloidal stable particles are obtained by this aggregation. In some embodiments, the particles comprise an amphiphilic lipid, particularly a cationic or cationic ionizable amphiphilic lipid, and a nucleic acid as described herein (e.g., DNA or RNA, e.g., mRNA). In some embodiments, the particles comprise a cationic / cationic ionizable lipid (e.g., a cationic ionizable lipid of the formula described herein), as well as at least one additional lipid, e.g., a neutral lipid (e.g., a phospholipid), an immunomodulator, a polymer-conjugated lipid, optionally a steroid (e.g., cholesterol), and combinations thereof, or consist of them. In some embodiments, the particles comprise a cationic polymer and a nucleic acid.

[0083] In some embodiments, the nucleic acid particles (e.g., ribonucleic acid particles) comprise more than one type of nucleic acid molecule, and the molecular parameters of the nucleic acid molecules may be similar or different from each other with respect to basic structural elements such as, for example, molar mass or molecular structure, capping, coding region, or other features.

[0084] In some embodiments, the nucleic acid particles described herein are nanoparticles. As used in this disclosure, "nanoparticle" refers to a particle having an average diameter suitable for parenteral administration and a diameter of less than 1000 nm. In some embodiments, the composition comprising the nanoparticles can have an average nanoparticle size (e.g., average diameter) of about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 70 to about 90 nm, or about 70 nm to about 80 nm. In some embodiments, the composition comprising the nanoparticles can have an average nanoparticle size (e.g., average diameter) of about 50 nm to about 100 nm. In some embodiments, the composition comprising the nanoparticles can have an average nanoparticle size (e.g., average diameter) of about 50 nm to about 150 nm. In some embodiments, the composition comprising the nanoparticles can have an average nanoparticle size (e.g., average diameter) of about 60 nm to about 120 nm. In some embodiments, the composition comprising the nanoparticles can have an average nanoparticle size (e.g., average diameter) of about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, or 150 nm.

[0085] A composition comprising the nucleic acid particles (e.g., ribonucleic acid particles) described herein can exhibit a polydispersity index of the nanoparticles of less than about 0.5, less than about 0.4, less than about 0.3, or about 0.2 or less. As an example, a composition comprising the nucleic acid particles (e.g., ribonucleic acid particles) described herein can exhibit a polydispersity index in the range of about 0.1 to about 0.3 or about 0.2 to about 0.3.

[0086] The nucleic acid particles (e.g., ribonucleic acid particles) described herein can be characterized by an "N / P ratio", which is the molar ratio of the cationic (nitrogen) groups in the cationic polymer ("N" in N / P) to the anionic (phosphate) groups in the RNA ("P" in N / P). The cationic groups are in the cationic form (e.g., N +) It is understood that it is either a group that is or a group that can be ionized to become cationic. The use of a single number in the N / P ratio (e.g., an N / P ratio of about 5) is intended to refer to that number being greater than 1. For example, an N / P ratio of about 4 is intended to mean about 4:1. In some embodiments, the nucleic acid particles described herein (e.g., ribonucleic acid particles) have an N / P ratio of 4 or more. In some embodiments, the nucleic acid particles described herein (e.g., ribonucleic acid particles) have an N / P ratio that is about 4 to about 12. In some embodiments, the nucleic acid particles described herein (e.g., ribonucleic acid particles) have an N / P ratio of about 4, 5, 6, 7, 8, 9, 10, 11, or 12. In some embodiments, the N / P ratio of the nucleic acid particles described herein (e.g., ribonucleic acid particles) is about 6.

[0087] The nucleic acid particles described herein (e.g., ribonucleic acid particles) can be prepared using a wide variety of methods that can include obtaining a colloid from at least one cationic lipid or cationic ionizable lipid or lipid-like material, and / or at least one cationic polymer, and mixing the colloid with a nucleic acid to obtain a nucleic acid particle. As used herein, an "ionizable" lipid, e.g., a "cationic ionizable" lipid or an "ionizable" polymer, e.g., a "cationic ionizable" polymer, can be a lipid or polymer that can be neutral at physiological pH in some embodiments, but can become cationic (i.e., positively charged) at neutral pH.

[0088] The term "average diameter" or "mean diameter" refers to the average hydrodynamic diameter of particles measured by dynamic light scattering (DLS) using data analysis that employs the so-called cumulative algorithm, which, as a result, provides a so-called Z-average having the dimension of length and a dimensionless polydispersity index (PDI) (Koppel, D., J. Chem. Phys. 57, 1972, pp 4814-4820, ISO 13321). Here, the "average diameter", "mean diameter", "diameter", or "size" of the particles is used synonymously with this value of the Z-average.

[0089] The "polydispersity index" is preferably calculated based on dynamic light scattering measurements by so-called cumulative analysis as described in the definition of "average diameter". Under certain preconditions, it can be taken as a measure of the overall size distribution of ribonucleic acid nanoparticles (e.g., ribonucleic acid nanoparticles).

[0090] It has been previously described that different types of nucleic acid particles are suitable for the delivery of nucleic acids in particulate form (e.g., Kaczmarek, J.C. et al., 2017, Genome Medicine 9, 60). For non-viral nucleic acid delivery vehicles, encapsulation of nucleic acids in nanoparticles can physically protect the nucleic acids from degradation and, depending on the particular chemical, assist in cell uptake and endosomal escape.

[0091] The present disclosure describes particles comprising a nucleic acid, at least one cationic lipid or a lipid that can be ionized cationically or a lipid-like material, and / or at least one cationic polymer that associates with the nucleic acid to form nucleic acid particles (e.g., ribonucleic acid particles, e.g., ribonucleic acid nanoparticles), and compositions comprising such particles. The nucleic acid particles (e.g., ribonucleic acid particles, e.g., ribonucleic acid nanoparticles) can comprise nucleic acids complexed in different forms by non-covalent interactions with the particles. In some embodiments described herein, the particles described herein are not viral particles, and in particular, they are not infectious viral particles, i.e., they cannot infect cells with a virus.

[0092] Some embodiments described herein relate to compositions, methods, and uses involving more than one, e.g., 2, 3, 4, 5, 6, or even more nucleic acid species.

[0093] In a nucleic acid particle (e.g., ribonucleic acid particle, e.g., ribonucleic acid nanoparticle) composition, each nucleic acid species can be formulated separately as an individual nucleic acid particle (e.g., ribonucleic acid particle, e.g., ribonucleic acid nanoparticle) formulation. In that case, each individual nucleic acid particle (e.g., ribonucleic acid particle, e.g., ribonucleic acid nanoparticle) formulation will contain one nucleic acid species. The individual nucleic acid particle (e.g., ribonucleic acid particle, e.g., ribonucleic acid nanoparticle) formulations can exist as separate entities, e.g., in separate containers. Such formulations are obtained by enabling the formation of particles by providing each nucleic acid species separately (typically in the form of a nucleic acid-containing solution) together with the particle-forming agent. Each particle will contain only the particular nucleic acid species provided when the particle is formed (individual particle formulation).

[0094] In some embodiments, a composition, e.g., a pharmaceutical composition, comprises more than one individual nucleic acid particle (e.g., ribonucleic acid particle, e.g., ribonucleic acid nanoparticle) formulation. Each pharmaceutical composition is referred to as a "mixed particle formulation". The mixed particle formulations according to the present invention are obtained by separately forming individual nucleic acid particle (e.g., ribonucleic acid particle, e.g., ribonucleic acid nanoparticle) formulations as described above and subsequently mixing the individual nucleic acid particle (e.g., ribonucleic acid particle, e.g., ribonucleic acid nanoparticle) formulations. By the mixing step, a formulation containing a mixed population of nucleic acid-containing particles is obtainable. The individual nucleic acid particle (e.g., ribonucleic acid particle, e.g., ribonucleic acid nanoparticle) populations may be together in one container, including the mixed population of the individual nucleic acid particle (e.g., ribonucleic acid particle, e.g., ribonucleic acid nanoparticle) formulations.

[0095] Alternatively, it is possible that various nucleic acid species are formulated together as a "combined particle formulation". Such a formulation is obtained by enabling the formation of particles by providing a mixed formulation (typically a mixed solution) of various nucleic acid species together with a particle-forming agent. In contrast to a "mixed particle formulation", a "combined particle formulation" will typically contain particles comprising more than one nucleic acid species. In a combined microparticle composition, the various nucleic acid species are typically present together within a single particle.

[0096] In certain embodiments, the nucleic acid, when present in the provided nucleic acid particles (e.g., ribonucleic acid particles, e.g., lipid nanoparticles, liposomes, polyplexes), is resistant to nuclease degradation in an aqueous solution.

[0097] Lipid nanoparticles In some embodiments, the nucleic acid particles (e.g., ribonucleic acid particles) are lipid nanoparticles. In some embodiments, the lipid nanoparticles are cationic lipid nanoparticles comprising one or more cationic lipids (e.g., those described herein), a nucleic acid (e.g., RNA), and an immunomodulatory agent. In some embodiments, the cationic lipid nanoparticles can comprise at least one cationic lipid, an immunomodulatory agent, at least one polymer-conjugated lipid, and at least one helper lipid. Lipid nanoparticles (LNPs) have proven useful for the delivery of nucleic acid cargo to target tissues. LNPs are used, for example, in certain commercially available vaccines for the treatment of COVID-19. However, some LNP formulations cause an inflammatory response in the body, such as an increase in cytokines and interleukins. This inflammatory response is associated with pain, swelling, fever, etc. However, the LNPs of the present disclosure do not suffer from the same defects associated with previous formulations.

[0098] The LNPs of the present disclosure comprise i) a cationic lipid, ii) a helper lipid, iii) a polymer-conjugated lipid (e.g., a polyethylene glycol-conjugated lipid "PEG lipid"), and iv) an immunomodulatory agent. In some embodiments, the LNPs described herein can further comprise additional additives described herein. The LNPs of the present disclosure can be useful in a variety of situations. For example, LNPs comprising a nucleic acid (e.g., RNA) described herein are useful for delivering the nucleic acid to target cells of a subject. In some embodiments, LNPs comprising a nucleic acid (e.g., RNA) described herein are useful for causing an increase in protein expression in a subject. In some embodiments, LNPs comprising a nucleic acid (e.g., RNA) described herein are useful for causing a pharmacological effect induced by protein expression in a subject. The lipid nanoparticles described herein are characterized by the molar percentage (mol%) of the components in the lipid nanoparticles. The mol% used with respect to the lipid components of the lipid nanoparticles is relative to the sum of the other lipid components in the lipid nanoparticles.

[0099] Cationic lipid As described herein, the LNPs of the present disclosure include cationic lipids. The cationic lipids described herein are lipids that are either positively charged or ionizable such that they become positively charged when exposed to certain physiological conditions, such as a pH of about 7.4 or less, and can promote lipid aggregation. In some embodiments, the cationic lipid is a lipid that contains one or more amine groups that have a positive charge or can have a positive charge (i.e., are ionizable).

[0100] In some embodiments, the cationic lipid is selected from 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (DMEPC); 2-dimyristoyl-3-trimethylammonium propane (DMTAP); dioleyl ether phosphatidylcholine (DOEPC); N,N-dioleyl-N,N-dimethylammonium chloride (DODAC); N-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA); N,N-distearyl-N,N-dimethylammonium bromide (DDAB); N-(2,3 dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP); 3-(N-(N′,N′dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), N-(1-(2,3-dioleyloxy)propyl)N-2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoroacetate (DOSPA), dioctadecylamidoglycyl carboxyspermine (DOGS), 1,2-dioleoyl-3-dimethylammonium propane (DODAP), N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), and N-(1,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethylammonium bromide (DMRIE).

[0101] In some embodiments, the cationic lipid is that provided in WO2012 / 016184, which is hereby incorporated by reference in its entirety. For example, in some embodiments, the cationic lipid is selected from 1,2-dilinoleyloxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-dilinoleyloxy-3-morpholinopropane (DLin-MA), 1,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), 1-linoleoyl-2-linoleoyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleyloxy-3-trimethylaminopropane chloride salt (DLin-TMA.Cl), 1,2-dilinoleoyl-3-trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2-dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), 3-(N,N dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-dioleylamino)-1,2-propanediol (DOAP), 1,2-dilinoleyl-oxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), and 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA).

[0102] In some embodiments, the cationic lipid is N,N-dimethyl-2,3-dioleyloxypropylamine (DODMA), 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA), 3-(N-(N′,N′-dimethylaminoethane)-carbamoyl) cholesterol (DC-Chol), dimethyldioctadecylammonium (DDAB); 1,2-dioleoyl-3-trimethylammonium propane (DOTAP); 1,2-dioleoyl-3-dimethylammonium-propane (DODAP); 1,2-acetyloxy-3-dimethylammonium propane; 1,2-dialkoxy-3-dimethylammonium propane; octadecyldimethylammonium chloride (DODAC), 1,2-distearyloxy-N,N-dimethyl-3-aminopropane (DSDMA), 2,3-di(tetradecyloxy)propyl-(2-hydroxyethyl)-dimethylazanium (DMRIE), 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (DMEPC), l,2-dimyristoyl-3-trimethylammonium propane (DMTAP), 1,2-dioleyloxypropyl-3-dimethyl-hydroxyethylammonium bromide (DORIE), and 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-l-propanamium trifluoroacetate (DOSPA), 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), dioctadecylamidoglycyl spermine (DOGS), 3-dimethylamino-2-(cholest-5-en-3-beta-oxybutan-4-oxy)-1-(cis,cis-9,12-octadecadienoxy)propane (CLinDMA), 2-[5′-(cholest-5-en-3-beta-oxy)-3′-oxapentoxy)-3-dimethyl-1-(cis,cis-9′,12′-octadecadienoxy)propane (CpLinDMA), N,N-dimethyl-3,4-dioleyloxybenzylamine (DMOBA), 1,2-N,N′-dioleylcarbamoyl-3-dimethylaminopropane (DOcarbDAP), 2,3-dilinoleoyloxy-N,N-dimethylpropylamine (DLinDAP), 1,2-N,N′-dilinoletylcarbamyl-3-dimethylaminopropane (DLincarbDAP), 1,2-dilinoleoylcarbamyl-3-dimethylaminopropane (DLinCDAP), 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA), 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-K-XTC2-DMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLin-KC2-DMA), heptatriaconta-6,9,28,31-tetraene-19-yl-4-(dimethylamino)butanoate (DLin-MC3-DMA), N-(2-hydroxyethyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)-1-propanaminium bromide (DMRIE), (±)-N-(3-aminopropyl)-N,N-dimethyl-2,3-bis(cis-9-tetradecenyl)oxy-1-propanaminium bromide (GAP-DMORIE), (±)-N-(3-aminopropyl)-N,N-dimethyl-2,3-bis(dodecyloxy)-1-propanaminium bromide (GAP-DLRIE), (±)-N-(3-aminopropyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)-1-propanaminium bromide (GAP-DMRIE), N-(2-aminoethyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)-1-propanaminium bromide (βAE-DMRIE), N-(4-carboxybenzyl)-N,N-dimethyl-2,3-bis(oleoyloxy)propane-1-aminium (DOBAQ), 2-({8-[(3β)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propane-1-amine (octyl-CLinDMA), 1,2-dimyristoyl-3-dimethylammonium-propane (DMDAP), 1,2-dipalmitoyl-3-dimethylammonium-propane (DPDAP), N1-[2-((1S)-1-[(3-amino-propyl)amino]-4-[di(3-aminopropyl)amino]butylcarboxamide)ethyl]-3,4-Dioleyloxybenzamide (MVL5), 1,2-Dioleoyl-sn-glycero-3-ethylphosphocholine (DOEPC), 2,3-Bis(dodecyloxy)-N-(2-hydroxyethyl)-N,N-dimethylpropan-1-ammonium bromide (DLRIE), N-(2-Aminoethyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)propan-1-ammonium bromide (DMORIE), Di((Z)-non-2-en-1-yl) 8,8'-(((2(dimethylamino)ethyl)thio)carbonyl)azanediyl) dioctanoate (ATX), N,N-Dimethyl-2,3-bis(dodecyloxy)propan-1-amine (DLDMA), N,N-Dimethyl-2,3-bis(tetradecyloxy)propan-1-amine (DMDMA), Di((Z)-non-2-en-1-yl)-9-((4-(dimethylaminobutanoyl)-oxy)heptadecanedioate (L319), N-Dodecyl-3-((2-dodecylcarbamoyl-ethyl)-{2-[(2-dodecylcarbamoyl-ethyl)-2-{(2-dodecylcarbamoyl-ethyl)-[2-(2-dodecylcarbamoyl-ethylamino)-ethyl]-amino}-ethyl-amino)propionamide (Lipidoid 98N, 12 -5), 1-[2-[Bis(2-hydroxydodecyl)amino]ethyl-[2-[4-[2-[Bis(2 hydroxydodecyl)amino]ethyl]piperazin-1-yl]ethyl]amino]dodecan-2-ol (Lipidoid C12-200), and the following structures (XV-1) to (XV-6):

Chemical formula

[0103] In some embodiments, the cationic lipid is provided in WO2021 / 026358, WO2020 / 219941, WO2017 / 075531, WO2016 / 118725, WO2016 / 118724, WO2016 / 176330, WO2017 / 049245, U.S. Patent No. 9,670,152, each of which is hereby incorporated by reference in its entirety.

[0104] In some embodiments, the cationic lipid is a compound of formula I: [Chemical formula] or a pharmaceutically acceptable salt thereof, wherein L 1 or L 2 One of is -OC(O)-, -C(O)O-, -C(O)-, -O-, -S(O) x -, -S-S-, -C(O)S-, SC(O)-, -NR a C(O)-, -C(O)NR a -, -NR a C(O)NR a -, -OC(O)NR a -, or -NR a C(O)O- and L 1 or L 2 The other of is -OC(O)-, -C(O)O-, -C(O)-, -O-, -S(O) x -, -S-S, -C(O)S-, SC(O)-, -NR a C(O)-, -C(O)NR a -, -NR a C(O)NR a -, -OC(O)NR a -, -NR a C(O)O-, or a direct bond, and G 1 and G 2 are each independently unsubstituted C 1 -C 12 alkylene or C 1 -C 12 alkenylene, and G 3 is C 1 -C 24 alkylene, C 1 -C 24 alkenylene, C 3 -C 8 cycloalkylene, C 3 -C 8 cycloalkenylene, and R a is H or C 1 -C12 is alkyl, R 1 and R 2 are each independently C 6 -C 24 alkyl or C 6 -C 24 alkenyl, R 3 is H, OR 5 , CN, -C(O)OR 4 , -OC(O)R 4 , or -R 5 C(O)R 4 and R 4 is C 1 -C 12 alkyl, R 5 is H or C 1 -C 6 alkyl, and x is 0, 1, or 2.

[0105] In some embodiments, one of L 1 or L 2 is -OC(O)- or -C(O)O-. In some embodiments, each of L 1 and L 2 is -OC(O)- or -C(O)O-.

[0106] In some embodiments, G 1 is C 1 -C 12 alkylene. In some embodiments, G 2 is C 1 -C 12 alkylene. In some embodiments, each of G 1 and G 2 is independently C 1 -C 12 alkylene. In some embodiments, each of G 1 and G 2 is independently C 5 -C 12 alkylene.

[0107] In some embodiments, G 3 is C 1 -C 24 alkylene. In some embodiments, G 3 is C 1 -C 6 alkylene.

[0108] In some embodiments, R 1 and R 2 are each independently

Chemical formula

[0109] In some embodiments, R 3 is OH.

[0110] In some embodiments, each of L 1 and L 2 is -OC(O)-, G 1 and G 2 are each independently C 5 -C 12 alkylene, G 3 is C 1 -C 6 alkylene, R 3 is OH, and R 1 and R 2 are each independently

Chemical formula

[0111] In some embodiments, the cationic lipid is a compound of formula Ia or Ib:

Chemical formula

[0112] In some embodiments, cationic lipids that may be useful according to the present disclosure are amino lipids that contain a titratable tertiary amino head group linked via ester bonds to at least two saturated alkyl chains, which ester bonds can be readily hydrolyzed to facilitate rapid degradation and / or excretion via the renal pathway. In some embodiments, such amino lipids have an apparent pK of about 6.0-6.5. a (e.g., an apparent pK of approximately 6.25) a In one embodiment, amino lipids are used in combination with LNPs, resulting in essentially entirely positively charged molecules at acidic pH (e.g., pH 5). In some embodiments, such amino lipids, when incorporated into LNPs, can impart distinct physicochemical properties that modulate particle formation, cellular uptake, fusogenicity, and / or endosomal release of the RNA(s). In some embodiments, introduction of an aqueous RNA solution to a lipid mixture containing such amino lipids at pH 4.0 can result in electrostatic interactions between the negatively charged RNA backbone and the positively charged cationic lipids. Without wishing to be bound by any particular theory, such electrostatic interactions result in efficient encapsulation of the RNA bulk and concomitant particle formation. After RNA encapsulation, adjustment of the pH of the medium surrounding the resulting LNPs to a more neutral pH (e.g., pH 7.4) results in neutralization of the surface charge of the LNPs. When all other variables are held constant, such charge-neutral particles exhibit longer in vivo circulation lifetimes and better delivery to hepatocytes compared to charged particles that are rapidly cleared by the reticuloendothelial system. Once inside the endosome, the low pH of the endosome renders LNPs containing such amino lipids fusogenic, allowing release of RNA into the cytoplasm of the target cell.

[0113] As described in this specification, the LNP contains at least one cationic lipid. In some embodiments, the cationic lipid is selected from Table 1 or

Table 1-1

Table 1-2

Table 1-3

Table 1-4

Table 1-5

Table 1-6

Table 1-7

Table 1-8

Table 1-9

Table 1-10

Table 1-11

Table 1-12

Table 1-13

Table 1-14

Table 1-15

[0114] In some embodiments, the cationic lipid is selected from Table 2, or [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] Or a pharmaceutically acceptable salt thereof. In some embodiments, the provided compound is provided and / or utilized in the form of a salt (e.g., a pharmaceutically acceptable salt). References to the compounds provided herein are understood to include references to their salts unless otherwise indicated.

[0115] In some embodiments, the cationic lipid is selected from Table 1 and / or 2.

[0116] In some embodiments, the cationic lipid is selected from DODMA, HY-501, ALC-0315, ALC366, and SM-102. In some embodiments, the cationic lipid is selected from ALC-0315 and ALC366. In some embodiments, the cationic lipid is ALC-0315. In some embodiments, the cationic lipid is ALC366. In some embodiments, the cationic lipid is SM-102. In some embodiments, the cationic lipid is DODMA. In some embodiments, the cationic lipid is HY-501.

[0117] In some embodiments, the LNP of the present disclosure contains about 30 to about 70 mol% of a cationic lipid relative to the total lipids in the LNP. In some embodiments, the LNP contains about 35 to about 65 mol% of a cationic lipid. In some embodiments, the LNP contains about 40 to about 60 mol% of a cationic lipid. In some embodiments, the LNP contains about 41 to about 49 mol% of a cationic lipid. In some embodiments, the LNP contains about 48 mol% of a cationic lipid. In some embodiments, the LNP contains about 50 mol% of a cationic lipid.

[0118] In some embodiments, the cationic ionizable lipid has the structure of formula (X) [Chemical formula] or a pharmaceutically acceptable salt, tautomer, prodrug, or stereoisomer thereof, wherein L 10 and L 20 One of them is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O) x’ -, -S-S-, -C(=O)S-, SC(=O)-, -NR a’ C(=O)-, -C(=O)NR a’ -, NR a’ C(=O)NR a’ -, -OC(=O)NR a’ -, or -NR a’is C(=O)O−, L 10 and L 20 the other of which is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -S-S-, -C(=O)S-, SC(=O)-, -NR a’ C(=O)-, -C(=O)NR a’ -, NR a’ C(=O)NR a’- -, -OC(=O)NR a’ -, or -NR a’ C(=O)O-, or a direct bond, G 1’ and G 2’ are each independently unsubstituted C 1 -C 12 alkylene or C 2-12 alkenylene, G 3’ is C 1-24 alkylene, C 2-24 alkenylene, C 3-8 cycloalkylene, or C 3-8 cycloalkenylene, R a’ is H or C 1-12 alkyl, R 35 and R 36 are each independently C 6-24 alkyl or C 6-24 alkenyl, R 37 is H, OR 50 , CN, -C(=O)OR 40 , -OC(=O)R 40 , or -NR 50 C(=O)R 40 wherein, R 40 is C 1-12 alkyl, R 50 is H or C 1-6 alkyl, x’ is 0, 1, or 2.

[0119] In some of the foregoing embodiments of formula (X), the lipid has the following structure (XA) or (XB): [Chemical formula] has one of the following, wherein R 35 , L 10 , G 1’ , G 2’ , L 20 , R 36 , R 37 , and R 60 are, both alone and in combination, as described in the classes and subclasses of this specification, A is a 3- to 8-membered cycloalkyl or cycloalkylene group, R 60 is, in each occurrence, independently H, OH, or C 1 -C 24 alkyl, n 1’ is an integer in the range of 1 to 15.

[0120] In some of the foregoing embodiments of formula (X), the lipid has structure (XA), and in other embodiments, the lipid has structure (XB).

[0121] In other embodiments of formula (X), the lipid has the following structure (XC) or (XD): [Chemical formula] has one of the following, wherein R 35 , L 10 , G 1’ , G 2’ , L 20 , R 36 , R 37 , and R 60 are, both alone and in combination, as described in the classes and subclasses of this specification, and y' and z' are each independently an integer in the range of 1 to 12.

[0122] In any of the foregoing embodiments of formula (X), L 10 and L 20One of them is -O(C=O)-. For example, in some embodiments, L 10 and L 20 each is -O(C=O)-. In some different embodiments of any of the foregoing, L 10 and L 20 are each independently -(C=O)O- or -O(C=O)-. For example, in some embodiments, L 10 and L 20 each is -(C=O)O-.

[0123] In some embodiments of formula (X), the lipid has one of the following structures (XE) or (XF):

Chemical formula

[0124] In some of the foregoing embodiments of formula (X), the lipid has one of the following structures (XG), (XH), (XJ), or (XK):

Chemical formula

[0125] In some of the foregoing embodiments of formula (X), n 1’ is an integer in the range of 2 to 12, for example, 2 to 8 or 2 to 4. For example, in some embodiments, n 1’is 3, 4, 5, or 6. In some embodiments, n 1’ is 3. In some embodiments, n 1’ is 4. In some embodiments, n 1’ is 5. In some embodiments, n 1’ is 6.

[0126] In some other embodiments of the foregoing embodiments of formula (X), y' and z' are each independently an integer in the range of 2 to 10. For example, in some embodiments, y' and z' are each independently an integer in the range of 4 to 9 or 4 to 6.

[0127] In some of the foregoing embodiments of formula (X), R 60 is H. In other embodiments of the foregoing embodiments, R 60 is C 1 -C 24 alkyl. In other embodiments, R 60 is OH.

[0128] In some embodiments of formula (X), G 3’ is unsubstituted. In other embodiments, G 3’ is substituted. In various different embodiments, G 3’ is linear C 1 -C 24 alkylene or linear C 2 -C 24 alkenylene.

[0129] In some other foregoing embodiments of formula (X), R 35 or R 36 , or both, are C 6 -C 24 alkenyl. For example, in some embodiments, R 35 and R 36 each independently have the following structure:

Chemical formula

[0130] In some of the foregoing embodiments of formula (X), at least one occurrence of R 7a is H. For example, in some embodiments, R 7a is H in each occurrence. In other different foregoing embodiments, at least one occurrence of R 7b is C 1 -C 8 alkyl. For example, in some embodiments, C 1 -C 8 alkyl is methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, n-hexyl, or n-octyl.

[0131] In different embodiments of formula (X), R 35 or R 36 , or both, have the following structure:

Chemical formula

[0132] In some of the foregoing embodiments of formula (X), R 37 is OH, CN, -C(=O)OR 40 , -OC(=O)R 40 , or -NHC(=O)R 40 . In some embodiments, R 40 is methyl or ethyl.

[0133] In various different embodiments, the cationic lipid of formula (X) has one of the structures shown below. [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula]

[0134] In various different embodiments, the cationic ionizable lipid has one of the structures shown in the following table. [Table 3]

[0135] In some embodiments, the cationic ionizable lipid has the structure of formula (XI): [Chemical formula] and wherein each of R 1” and R 2”” is independently R 5” or -G 1” -L 1” -R 6” and at least one of R 1” and R 2” is -G 1” -L 1” -R 6” and R 3” and R 4” each independently is a C 1-6 alkyl, C 2-6 alkenyl, aryl, and C 3-10 selected from the group consisting of cycloalkyl, R 5” and R 6” each independently is an acyclic hydrocarbyl group having at least 10 carbon atoms, G 1” and G2” each independently is an unsubstituted C 1-12 alkylene or C 2-12 alkenylene, L 1” and L 2” each independently is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O) x” -, -S-S-, -C(=O)S-, -SC(=O)-, -NR a” C(=O)-, -C(=O)NR a” -, -NR a” C(=O)NR a”- -, -OC(=O)NR a” -, and -NR a” C(=O)O- selected from the group consisting of, R a” is H or C 1-12 alkyl, “m” is 0, 1, 2, 3, or 4, “x” is 0, 1, or 2.

[0136] In some of the foregoing embodiments of formula (XI), G 1” each independently is an unsubstituted C 1 -C 12 alkylene or unsubstituted C 2-12 alkenylene, for example, unsubstituted, straight-chain C 1-12 alkylene or unsubstituted, straight-chain C 2-12 alkenylene. In some embodiments, each G 1” each independently is an unsubstituted C 6-12 alkylene or unsubstituted C 6-12An alkenylene, for example, unsubstituted, straight-chain C 6-12 alkylene or unsubstituted, straight-chain C 6-12 alkenylene. In some embodiments, each G 1” is, independently, unsubstituted C 8-12 alkylene or unsubstituted C 8-12 alkenylene, for example, unsubstituted, straight-chain C 8-12 alkylene or unsubstituted, straight-chain C 8-12 alkenylene. In some embodiments, each G 1” is, independently, unsubstituted C 6-10 alkylene or unsubstituted C 6-10 alkenylene, for example, unsubstituted, straight-chain C 6-10 alkylene or unsubstituted, straight-chain C 6-10 alkenylene. In some embodiments, each G 1” is, independently, unsubstituted alkylene having 8, 9, or 10 carbon atoms, for example, unsubstituted, straight-chain alkylene having 8, 9, or 10 carbon atoms. In some embodiments, R 1” and R 2” are both, independently, -G 1” -L 1” -R 6” wherein G 1” for R 1” can be different from G 2” for R 1” . In some of these embodiments, for example, G 1” for R 1” is unsubstituted, straight-chain C 1-12 alkylene and G 2” for R 1” is unsubstituted, straight-chain C 2-12 alkenylene or G 1” for R 1” is unsubstituted, straight-chain C 1-12 alkylene group and G 2” for R 1” is a different unsubstituted, straight-chain C 1-12 alkylene group. In some embodiments, R 1” and R 2” are both, independently, -G 1” -L1” -R 6” and R 1” for G 1” is the same as R 2” for G 1” may be the same. In some of these embodiments, for example, each G 1” is the same unsubstituted, straight-chain C 8-12 alkylene, for example, unsubstituted, straight-chain C 8-10 alkylene, or each G 1” is the same unsubstituted, straight-chain C 6-12 alkenylene.

[0137] In some of the foregoing embodiments of formula (XI), each L 1” is independently selected from the group consisting of -O(C=O)-, -(C=O)O-, -C(=O)S-, -SC(=O)-, -NR a” C(=O)-, and -C(=O)NR a” -. In some embodiments, the R 1” of L a” is H or C 1-12 alkyl. In some embodiments, the R 1” of L a” is H or C 1-6 alkyl, for example, H or C 1-3 alkyl. In some embodiments, the R 1” of L a” is H, methyl, or ethyl. In some embodiments, each L 1” is independently selected from the group consisting of -O(C=O)-, -(C=O)O-, -C(=O)S-, and -SC(=O)-. In some embodiments, each L 1” is independently -O(C=O)- or -(C=O)O-. In some embodiments, R 1” and R 2” are both independently -G 1” -L 1” -R 6” and the L 1” for R 1” is the L 2” for R 1”can be different. In some of these embodiments, for example, R 1” for L 1” is one selected from the group consisting of -O(C=O)-, -(C=O)O-, -C(=O)S-, -SC(=O)-, -NR a” C(=O)-, and -C(=O)NR a” - (for example, L 1” for R 1” is -O(C=O)-), L 2” for R 1” is a different one selected from the group consisting of -O(C=O)-, -(C=O)O-, -C(=O)S-, -SC(=O)-, -NR a” C(=O)-, and -C(=O)NR a” - (for example, L 2” for R 1” is -(C=O)O-). In some embodiments, both R 1” and R 2” are each independently -G 1” -L 1” -R 6” and L 1” for R 1” can be the same as L 2” for R 1” In some of these embodiments, for example, each L 1” is the same one selected from the group consisting of -O(C=O)-, -(C=O)O-, -C(=O)S-, -SC(=O)-, -NR a” C(=O)-, and -C(=O)NR a”- -; for example, each L 1” is -O(C=O)- or each L 1” is -(C=O)O-.

[0138] In some of the foregoing embodiments of formula (XI), each R 6” is independently an acyclic hydrocarbyl group having at least 10 carbon atoms, for example, a straight-chain hydrocarbyl group having at least 10 carbon atoms. In some embodiments, each R 6”independently has up to 30 carbon atoms, such as up to 28, up to 26, up to 24, up to 22, or up to 20 carbon atoms. In some embodiments, each R 6” independently is an acyclic hydrocarbyl group having 10 to 30 carbon atoms (such as 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), for example, a linear hydrocarbyl group having 10 to 30 carbon atoms (such as 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms). In some embodiments, each R 6” is R 6” is bonded to L 1” through the internal carbon atom of R 6” independently has up to 30 carbon atoms (such as up to 28, up to 26, up to 24, up to 22, or up to 20 carbon atoms), and each R 6” is R 6” is bonded to L 1” through the internal carbon atom of R 6” independently is an acyclic hydrocarbyl group having at least 10 carbon atoms, for example, a linear hydrocarbyl group having at least 10 carbon atoms, and each R 6” is R 6” is bonded to L 1” through the internal carbon atom of R 6” independently is an acyclic hydrocarbyl group having 10 to 30 carbon atoms (such as 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), for example, a linear hydrocarbyl group having 10 to 30 carbon atoms (such as 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), and each R 6” is R 6” is bonded to L 1” through the internal carbon atom of R 6” The hydrocarbyl group of is an alkyl or alkenyl group, for example, a C 10-30 alkyl or alkenyl group. Thus, in some embodiments, each R6” is, independently, an acyclic alkyl group having at least 10 carbon atoms, or an acyclic alkenyl group having at least 10 carbon atoms, for example, a linear alkyl group having at least 10 carbon atoms, or a linear alkenyl group having at least 10 carbon atoms. In some embodiments, each R 6” is, independently, an acyclic alkyl group having 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), or an acyclic alkenyl group having 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), for example, a linear alkyl group having 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), or a linear alkenyl group having 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms). In some embodiments, each R 6” is, independently, an acyclic alkyl group having 11 to 19 carbon atoms (for example, 11, 13, 15, 17, or 17 carbon atoms), for example, a linear alkyl group having 11 to 19 carbon atoms (for example, 11, 13, 15, 17, or 17 carbon atoms). In some embodiments, each R 6” is, independently, an acyclic alkyl group having 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), or an acyclic alkenyl group having 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), for example, a linear alkyl group having 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), or a linear alkenyl group having 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms), and each R 6” is R 6” is bonded to L 1” through the internal carbon atoms of. In some embodiments, each R6” is, independently, an acyclic alkyl group having 11 to 19 carbon atoms (e.g., 11, 13, 15, 17, or 17 carbon atoms), for example, a linear alkyl group having 11 to 19 carbon atoms (e.g., 11, 13, 15, 17, or 17 carbon atoms), and each R 6” is R 6” is bonded to L through an internal carbon atom of R 1” . The expression "internal carbon atom" means that the carbon atom of R 6” to which R 1” is bonded to L 6” is directly bonded to at least two other carbon atoms of R 6” . For example, for the following C 11 alkyl group, each carbon atom at any one of positions 2, 3, 4, 5, and 7 is eligible as an "internal carbon atom" according to the present disclosure, but the carbon atoms at positions 1, 6, 8, 9, 10, and 11 are not eligible.

Chemical formula

[0139] Therefore, R 6” is a C 1” alkyl group bonded to L through the internal carbon of R 11 and is the following group: 6”

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0140] In some of the foregoing embodiments of formula (XI), R 5” is an acyclic hydrocarbyl group having at least 10 carbon atoms, e.g., a linear hydrocarbyl group having at least 10 carbon atoms. In some embodiments, R 5” is an acyclic hydrocarbyl group having at least 12 carbon atoms, e.g., a linear hydrocarbyl group having at least 12, at least 14, at least 16, or at least 18 carbon atoms. In some embodiments, R 5” has up to 30 carbon atoms, e.g., up to 28, up to 26, up to 24, up to 22, or up to 20 carbon atoms. In some embodiments, R 5”is an acyclic hydrocarbyl group, for example, a straight-chain hydrocarbyl group, and each hydrocarbyl group has 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, 10 to 20 carbon atoms, or 12 to 30, 12 to 28, 12 to 26, 12 to 24, 12 to 22, 12 to 20 carbon atoms, or 14 to 30, 14 to 28, 14 to 26, 14 to 24, 14 to 22, 14 to 20 carbon atoms, or 16 to 30, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms). In some embodiments, R 5” 's hydrocarbyl group is an alkyl or alkenyl group, for example, a C 10-30 alkyl or alkenyl group. Thus, in some embodiments, R 5” is an acyclic alkyl group having at least 10 carbon atoms (for example, at least 12, at least 14, at least 16, or at least 18 carbon atoms), or an acyclic alkenyl group having at least 10 carbon atoms (at least 12, at least 14, at least 16, or at least 18 carbon atoms), for example, a straight-chain alkyl group having at least 10 carbon atoms (at least 12, at least 14, at least 16, or at least 18 carbon atoms), or a straight-chain alkenyl group having at least 10 carbon atoms (at least 12, at least 14, at least 16, or at least 18 carbon atoms). In some embodiments, R 5”is an acyclic alkyl group or an acyclic alkenyl group, for example, a straight-chain alkyl group or a straight-chain alkenyl group, and each of the alkyl group and the alkenyl group independently has 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, 10 to 20 carbon atoms, or 12 to 30, 12 to 28, 12 to 26, 12 to 24, 12 to 22, 12 to 20 carbon atoms, or 14 to 30, 14 to 28, 14 to 26, 14 to 24, 14 to 22, 14 to 20 carbon atoms, or 16 to 30, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms). In some embodiments, the alkenyl group has at least 2 carbon-carbon double bonds, for example, 2 or 3 carbon-carbon double bonds, for example, 2 carbon-carbon double bonds. In some embodiments, the alkenyl group has at least 1 carbon-carbon double bond in a cis configuration, for example, 1, 2, or 3, for example, 2 carbon-carbon double bonds in a cis configuration. Thus, R 5” is an acyclic alkyl group or an acyclic alkenyl group, for example, a straight-chain alkyl group or a straight-chain alkenyl group, and each of the alkyl group and the alkenyl group independently has 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, 10 to 20 carbon atoms, or 12 to 30, 12 to 28, 12 to 26, 12 to 24, 12 to 22, 12 to 20 carbon atoms, or 14 to 30, 14 to 28, 14 to 26, 14 to 24, 14 to 22, 14 to 20 carbon atoms, or 16 to 30, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms), and the alkenyl group has at least 2 carbon-carbon double bonds, for example, 2 or 3 carbon double bonds. In some embodiments, R 5”is an acyclic alkyl group or an acyclic alkenyl group, for example, a linear alkyl group or a linear alkenyl group, and each of the alkyl group and the alkenyl group independently has 10 to 30 carbon atoms (for example, 10 to 28, 10 to 26, 10 to 24, 10 to 22, 10 to 20 carbon atoms, or 12 to 30, 12 to 28, 12 to 26, 12 to 24, 12 to 22, 12 to 20 carbon atoms, or 14 to 30, 14 to 28, 14 to 26, 14 to 24, 14 to 22, 14 to 20 carbon atoms, or 16 to 30, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms), and the alkenyl group has at least one carbon-carbon double bond, for example, 1, 2, or 3 carbon-carbon double bonds, in a cis configuration. In some embodiments, R 5” is the following structure:

Chemical formula

Chemical formula

[0141] In some of the foregoing embodiments of formula (XI), L 2” is selected from the group consisting of -O(C=O)-, -(C=O)O-, -C(=O)-, -S-S-, -C(=O)S-, -SC(=O)-, -NR a” C(=O)-, -C(=O)NR a” -, -NR a” C(=O)NR a” -, -OC(=O)NR a” -, and -NR a” C(=O)O-. In some embodiments, L 2” is -O(C=O)-, -(C=O)O-, -C(=O)-, -C(=O)S-, -SC(=O)-, -NR a” C(=O)-, and -C(=O)NR a”is selected from the group consisting of. In some embodiments, L 2” of R a” is H or C 1-12 alkyl. In some embodiments, L 2” of R a” is H or C 1-6 alkyl, for example, H or C 1-3 alkyl. In some embodiments, L 2” of R a” is H, methyl, or ethyl. In some embodiments, L 2” is selected from the group consisting of -O(C=O)-, -(C=O)O-, -C(=O)S-, and -SC(=O). In some embodiments, L 2” is -O(C=O)- or -(C=O)O-.

[0142] In some of the foregoing embodiments of formula (XI), G 2” is unsubstituted C 1-12 alkylene or unsubstituted C 2-12 alkenylene, for example, unsubstituted, straight-chain C 1-12 alkylene or unsubstituted, straight-chain C 2-12 alkenylene. In some embodiments, G 2” is unsubstituted C 2-10 alkylene or unsubstituted C 2-10 alkenylene, for example, unsubstituted, straight-chain C 2-10 alkylene or unsubstituted, straight-chain C 2-10 alkenylene. In some embodiments, G 2” is unsubstituted C 2-6 alkylene or unsubstituted C 2-6 alkenylene, for example, unsubstituted, straight-chain C 2-6 alkylene or unsubstituted, straight-chain C 2-6 alkenylene. In some embodiments, G 2” is unsubstituted C 2-4 alkylene or unsubstituted C 2-4 alkenylene, for example, unsubstituted, straight-chain C 2-4 alkylene or unsubstituted, straight-chain C 2-4 alkenylene. In some embodiments, G 2”is ethylene or trimethylene.

[0143] In some of the foregoing embodiments of formula (XI), R 3” and R 4” each independently is C 1-6 alkyl or C 2-6 alkenyl. In some embodiments, R 3” and R 4” each independently is C 1-4 alkyl or C 2-4 alkenyl. In some embodiments, R 3” and R 4” each independently is C 1-3 alkyl. In some embodiments, R 3” and R 4” each independently is methyl or ethyl. In some embodiments, R 3” and R 4” each is methyl.

[0144] In some of the foregoing embodiments of formula (XI), m'' is 0, 1, 2, or 3. In some embodiments, m'' is 0 or 2. In some embodiments, m'' is 0. In some embodiments, m'' is 2.

[0145] In some of the foregoing embodiments of formula (XI), the cationic ionizable lipid has a structure of formula (XIIa) or (XIIb),

Chemical Formula

[0146] In some of the foregoing embodiments of formula (XIIa), R 5”has up to 30 carbon atoms, for example, up to 28, up to 26, up to 24, up to 22, or up to 20 carbon atoms. In some embodiments of formula (XIIa), R 5” is a straight-chain hydrocarbyl group having from 14 to 30 carbon atoms (e.g., from 14 to 28, from 14 to 26, from 14 to 24, from 14 to 22, from 14 to 20 carbon atoms, or from 16 to 30, from 16 to 28, from 16 to 26, from 16 to 24, from 16 to 22, from 16 to 20 carbon atoms, or from 18 to 30, from 18 to 28, from 18 to 26, from 18 to 24, from 18 to 22, or from 18 to 20 carbon atoms). In some embodiments of formula (XIIa), R 5” is a straight-chain alkyl or alkenyl group having from 14 to 30 carbon atoms (e.g., from 14 to 28, from 14 to 26, from 14 to 24, from 14 to 22, from 14 to 20 carbon atoms, or from 16 to 30, from 16 to 28, from 16 to 26, from 16 to 24, from 16 to 22, from 16 to 20 carbon atoms, or from 18 to 30, from 18 to 28, from 18 to 26, from 18 to 24, from 18 to 22, or from 18 to 20 carbon atoms). In some embodiments of formula (XIIa), the alkenyl group has at least 2 carbon-carbon double bonds, for example, 2 or 3 carbon-carbon double bonds, for example, 2 carbon-carbon double bonds. In some embodiments, the alkenyl group has at least 1 carbon-carbon double bond in a cis configuration, for example, 1, 2, or 3, for example, 2 carbon-carbon double bonds in a cis configuration. Thus, in some embodiments of formula (XIIa), R 5” is a straight-chain alkyl group or a straight-chain alkenyl group, each of the alkyl group and the alkenyl group independently having from 14 to 30 carbon atoms (e.g., from 14 to 28, from 14 to 26, from 14 to 24, from 14 to 22, from 14 to 20 carbon atoms, or from 16 to 30, from 16 to 28, from 16 to 26, from 16 to 24, from 16 to 22, from 16 to 20 carbon atoms, or from 18 to 30, from 18 to 28, from 18 to 26, from 18 to 24, from 18 to 22, or from 18 to 20 carbon atoms), and the alkenyl group has at least 2 carbon-carbon double bonds, for example, 2 or 3 carbon-carbon double bonds. In some embodiments of formula (XIIa), R 5”is a linear alkyl group or a linear alkenyl group, each of the alkyl group and the alkenyl group independently having 14 to 30 carbon atoms (for example, 14 to 28, 14 to 26, 14 to 24, 14 to 22, 14 to 20 carbon atoms, or 16 to 30, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms), the alkenyl group having at least one carbon-carbon double bond, for example, 1, 2, or 3 carbon-carbon double bonds, in a cis configuration. In some embodiments of formula (XIIa), R 5” is a linear alkyl group or a linear alkenyl group, each of the alkyl group and the alkenyl group independently having 14 to 30 carbon atoms (for example, 14 to 28, 14 to 26, 14 to 24, 14 to 22, 14 to 20 carbon atoms, or 16 to 30, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms), the alkenyl group having 2 or 3 carbon-carbon double bonds, and at least one carbon-carbon double bond, for example 1, 2, or 3 carbon-carbon double bonds, being in a cis configuration. In some embodiments of formula (XIIa), R 5” has the following structure:

Chemical formula

Chemical formula

[0147] In some of the foregoing embodiments of formula (XIIb), each R 6” is, independently, up to 30 carbon atoms, for example, up to 28, up to 26, up to 24, up to 22, or up to 20 carbon atoms. In some embodiments of formula (XIIb), each R 6” is, independently, a straight-chain hydrocarbyl group (for example, a straight-chain alkyl group) having from 10 to 30 carbon atoms (for example, from 10 to 28, from 10 to 26, from 10 to 24, from 10 to 22, or from 10 to 20 carbon atoms, or from 11 to 19 carbon atoms, for example, 11, 13, 15, 17, or 17 carbon atoms). In some embodiments of formula (XIIb), each R 6” is, independently, a straight-chain hydrocarbyl group (for example, a straight-chain alkyl group) having from 10 to 30 carbon atoms (for example, from 10 to 28, from 10 to 26, from 10 to 24, from 10 to 22, or from 10 to 20 carbon atoms, or from 11 to 19 carbon atoms, for example, 11, 13, 15, 17, or 17 carbon atoms), and each R 6” is bonded to L 6” through an internal carbon atom of R 1” . In some embodiments of formula (XIIb), each R 6” is, independently,

Chemical formula

Chemical formula

[0148] In some of the aforementioned embodiments of formula (XI), the cationic ionizable lipid has a structure of formula (XIIIa) or (XIIIb),

Chemical Structure

[0149] In some of the foregoing embodiments of formula (XIIIa), R 5” has up to 30 carbon atoms, for example, up to 28, up to 26, up to 24, up to 22, or up to 20 carbon atoms. In some embodiments of formula (XIIIa), R 5” is a straight-chain alkyl or alkenyl group having 16 to 30 carbon atoms (for example, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms). In some embodiments of formula (XIIIa), the alkenyl group has at least 2 carbon-carbon double bonds, for example, 2 or 3 carbon-carbon double bonds, for example, 2 carbon-carbon double bonds. In some embodiments, the alkenyl group has at least 1 carbon-carbon double bond in the cis configuration, for example, in the cis configuration, 1, 2, or 3, for example, 2 carbon-carbon double bonds. Thus, in some embodiments of formula (XIIIa), R 5”is a linear alkyl group or a linear alkenyl group, and each of the alkyl group and the alkenyl group independently has 16 to 30 carbon atoms (for example, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms), and the alkenyl group has at least 2 carbon-carbon double bonds, for example, 2 or 3 carbon double bonds. In some embodiments of formula (XIIIa), R 5” is a linear alkyl group or a linear alkenyl group, and each of the alkyl group and the alkenyl group independently has 16 to 30 carbon atoms (for example, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms), and the alkenyl group has at least 1 carbon-carbon double bond in a cis configuration, for example, 1, 2, or 3 carbon-carbon double bonds. In some embodiments of formula (XIIIa), R 5” is a linear alkyl group or a linear alkenyl group, and each of the alkyl group and the alkenyl group independently has 16 to 30 carbon atoms (for example, 16 to 28, 16 to 26, 16 to 24, 16 to 22, 16 to 20 carbon atoms, or 18 to 30, 18 to 28, 18 to 26, 18 to 24, 18 to 22, or 18 to 20 carbon atoms), and the alkenyl group has 2 or 3 carbon-carbon double bonds, and at least 1 carbon-carbon double bond, for example, 1, 2, or 3 carbon-carbon double bonds, is in a cis configuration. In some embodiments of formula (XIIIa), R 5” has the following structure:

Chemical formula

Chemical formula

[0150] In some of the foregoing embodiments of formula (XIIIb), each R 6” is independently a hydrocarbyl group having up to 30 carbon atoms, such as up to 28, up to 26, up to 24, up to 22, or up to 20 carbon atoms. In some embodiments of formula (XIIIb), each R 6” is independently a straight-chain hydrocarbyl group (e.g., a straight-chain alkyl group) having 10 to 30 carbon atoms (e.g., 10 to 28, 10 to 26, 10 to 24, 10 to 22, or 10 to 20 carbon atoms, or 11 to 19 carbon atoms, such as 11, 13, 15, 17, or 17 carbon atoms), and each R 6” is bonded to L 6” through an internal carbon atom of R 1” . In some embodiments of formula (XIIIb), each R 6” is bonded to L 6” through an internal carbon atom of R 1” and is independently selected from the group consisting of

Chemical formula

Chemical formula

[0151] In some of the foregoing embodiments of formula (XI), the cationic ionizable lipid is of the following formulas (XIV-1), (XIV-2), and (XIV-3):

Chemical formula

[0152] In some embodiments, the cationic ionizable lipid is (6Z,16Z)-12-((Z)-dec-4-en-1-yl)docosa-6,16-dien-11-yl 5-(dimethylamino)pentanoate (3D-P-DMA). The structure of 3D-P-DMA can be represented as follows:

Chemical formula

[0153] In various different embodiments, the cationic ionizable lipid is selected from the group consisting of N,N-dimethyl-2,3-dioleyloxypropylamine (DODMA), 1,2-dioleoyl-3-dimethylammonium-propane (DODAP), heptatriaconta-6,9,28,31-tetraene-19-yl-4-(dimethylamino)butanoate (DLin-MC3-DMA), and 4-((di((9Z,12Z)-octadeca-9,12-dien-1-yl)amino)oxy)-N,N-dimethyl-4-oxobutane-1-amine (DPL-14).

[0154] Further examples of cationic ionizable lipids include 3-(N-(N′,N′-dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), 1,2-dioleoyl-3-dimethylammonium-propane (DODAP); 1,2-diacetyloxy-3-dimethylammonium propane; 1,2-dialkoxy-3-dimethylammonium propane, 1,2-distearyloxy-N,N-dimethyl-3-aminopropane (DSDMA), 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), dioctadecylamidoglycyl spermine (DOGS), 3-dimethylamino-2-(cholest-5-en-3-beta-oxybutane-4-oxy)-1-(cis,cis-9,12-octadecadienoxy)propane (CLinDMA), 2-[5′-(cholest-5-en-3-beta-oxy)-3′-oxapentoxy)-3-dimethyl-1-(cis,cis-9′,12′-octadecadienoxy)propane (CpLinDMA), N,N-dimethyl-3,4-dioleyloxybenzylamine (DMOBA), 1,2-N,N′-dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), 2,3-dilinoleyloxy-N,N-dimethylpropylamine (DLinDAP), 1,2-N,N′-dilinooleylcarbamyl-3-dimethylaminopropane (DLincarbDAP), 1,2-dilinoleylcarbamyl-3-dimethylaminopropane (DLinCDAP), 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA), 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-K-XTC2-DMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLin-KC2-DMA), heptatriaconta-6,9,28,31-tetraene-19-yl-4-(dimethylamino)butanoate (DLin-MC3-DMA), 2-({8-[(3β)-cholest-5-en-3-yl-oxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-1-amine (Octyl-CLinDMA), 1,2-dimyristoyl-3-dimethylammonium-propane (DMDAP), 1,2-dipalmitoyl-3-dimethylammonium-propane (DPDAP), N1-[2-((1S)-1-[(3-aminopropyl)amino]-4-[di(3-aminopropyl)amino]butylcarboxamide)ethyl]-3,4-di[oleyloxy]benzamide (MVL5), bis((Z)-non-2-en-1-yl) 8,8’-((((2(dimethylamino)ethyl)thio)carbonyl)azanediyl)dioctanoate (ATX), N,N-dimethyl-2,3-bis(dodecyloxy)propan-1-amine (DLDMA), N,N-dimethyl-2,3-bis(tetradecyloxy)propan-1-amine (DMDMA), bis((Z)-non-2-en-1-yl)-9-((4-(dimethylaminobutanoyl)oxy)heptadecanedioate (L319), N-dodecyl-3-((2-dodecylcarbamoyl-ethyl)-{2-[(2-dodecylcarbamoyl-ethyl)-2-{(2-dodecylcarbamoyl-ethyl)-[2-(2-dodecylcarbamoyl-ethylamino)-ethyl]-amino}-ethylamino)propionamide (Lipidoid 98N12-5), 1-[2-[bis(2-hydroxydodecyl)amino]ethyl-[2-[4-[2-[bis(2 hydroxydodecyl)amino]ethyl]piperazin-1-yl]ethyl]amino]dodecan-2-ol (Lipidoid C12-200) are included, but are not limited thereto.,

[0155] In certain embodiments, the cationic ionizable lipid is, or comprises, X-3.

[0156] In certain embodiments, the cationic ionizable lipid is, or comprises, X-45.

[0157] In some embodiments, the cationic lipid for use herein is, or comprises, DPL-14. As used herein, “DPL-14” has the following general formula: [Chemical formula] comprises

[0158] In some embodiments, the cationic lipid for use herein is, or comprises, EA-2. As used herein, "EA-2" has the following general formula: [Chemical formula] comprises

[0159] It should be understood that any reference to a cationic lipid or cationic ionizable lipid disclosed herein also includes its salts (especially pharmaceutically acceptable salts), tautomers, stereoisomers, solvates (e.g., hydrates), and isotopically labeled forms.

[0160] In some embodiments, the cationic / cationic ionizable lipid can be included at about 10 mol% to about 100 mol%, about 20 mol% to about 100 mol%, about 30 mol% to about 100 mol%, about 40 mol% to about 100 mol%, or about 50 mol% to about 100 mol% of the total lipids present in the composition / particles. In some embodiments, the cationic / cationic ionizable lipid is included at about 40 mol% to about 75 mol%, preferably about 40 mol% to about 70 mol%, more preferably about 45 mol% to about 65 mol% of the total lipids present in the composition / particles.

[0161] In some embodiments, the nucleic acid compositions (especially DNA or RNA compositions) described herein comprise a cationic lipid or cationic ionizable lipid and one or more additional lipids, and the cationic lipid or cationic ionizable lipid is included at about 10 mol% to about 80 mol%, about 20 mol% to about 75 mol%, about 20 mol% to about 70 mol%, about 20 mol% to about 60 mol%, about 25 mol% to about 55 mol%, about 30 mol% to about 50 mol%, about 35 mol% to about 45 mol%, or about 40 mol% to about 55 mol% of the total lipids present in the composition.

[0162] In some embodiments of the nucleic acid (e.g., DNA or RNA) compositions (particularly, mRNA compositions) described herein, (i) the nucleic acid, and (ii) at least a portion of a cationic lipid or cationic ionizable lipid form particles (e.g., LNPs), and the cationic lipid or cationic ionizable lipid may be present in the particles at about 10 mol% to about 80 mol%, about 20 mol% to about 75 mol%, about 20 mol% to about 70 mol%, about 20 mol% to about 60 mol%, about 25 mol% to about 55 mol%, about 30 mol% to about 50 mol%, about 35 mol% to about 45 mol%, or about 40 mol% to about 55 mol% of the total lipids present in the particles.

[0163] Helper lipid As described herein, the LNPs of the present disclosure contain helper lipids. In some embodiments, the helper lipid is a phospholipid. In some embodiments, the helper lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), phosphatidylethanolamine, e.g., 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), sphingomyelin (SM), 1,2-diacylglyceryl-3-O-4'-(N,N,N-trimethyl)-homoserine (DGTS), ceramide, cholesterol, a steroid, e.g., a sterol, and derivatives thereof, or includes them.

[0164] In some embodiments, the helper lipid is, or comprises, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, or sphingomyelin. In some embodiments, the helper lipid is diacyl phosphatidylcholine, such as distearoyl phosphatidylcholine (DSPC), dioleoyl phosphatidylcholine (DOPC), dimyristoyl phosphatidylcholine (DMPC), dipalmitoyl phosphatidylcholine (DPPC), dilauroyl phosphatidylcholine (DLPC), dioleoyl phosphatidylethanolamine (DOPE), distearoyl phosphatidylethanolamine (DSPE), dipalmitoyl phosphatidylethanolamine (DPPE), dimyristoyl phosphatidylethanolamine (DMPE), dilauroyl phosphatidylethanolamine (DLPE), diphytanoyl phosphatidylethanolamine (DPyPE), 1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine (DOPG), 1,2-dipalmitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DPPG), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), N-palmitoyl-D-erythro-sphingosylphosphorylcholine (SM), or comprises the same.In some embodiments, the helper lipid is selected from the group consisting of DSPC, DOPC, DMPC, DPPC, POPC, DOPE, DOPG, DPPG, POPE, DPPE, DMPE, DSPE, and SM. In some embodiments, the neutral lipid is selected from the group consisting of DSPC, DPPC, DMPC, DOPC, POPC, DOPE, and SM. In some embodiments, the neutral lipid is DSPC.

[0165] The helper lipid can be synthetic or of natural origin. Other helper lipids suitable for use in lipid nanoparticles are described in WO2021 / 026358, WO2017 / 075531, and WO2018 / 081480, the entire contents of each of which are hereby incorporated by reference in their entirety.

[0166] In some embodiments, the lipid nanoparticles contain about 5 to about 15 mol% phospholipid. In some embodiments, the lipid nanoparticles contain about 8 to about 12 mol% phospholipid. In some embodiments, the lipid nanoparticles contain about 10 mol% phospholipid. In some embodiments, the lipid nanoparticles contain about 5 to about 15 mol% DSPC. In some embodiments, the lipid nanoparticles contain about 8 to about 12 mol% DSPC. In some embodiments, the lipid nanoparticles contain about 10 mol% DSPC.

[0167] Polymer-conjugated lipid As described herein, the LNPs of the present disclosure include polymer-conjugated lipids. In some embodiments, the polymer-conjugated lipid is a lipid conjugated with polyethylene glycol (PEG lipid). In some embodiments, the PEG lipid is selected from pegylated diacylglycerol (PEG-DAG), such as l-(monomethoxy-polyethylene glycol)-2,3-dimyristoyl glycerol (PEG-DMG) (e.g., 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (PEG2000-DMG)), pegylated phosphatidylethanolamine (PEG-PE), PEG succinate diacylglycerol (PEG-S-DAG), such as 4-O-(2’,3’-di(tetradecanoyloxy)propyl-1-O-(ω-methoxy(polyethoxy)ethyl)butanedioate (PEG-S-DMG), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG2000 amine), pegylated ceramide (PEG-cer), or PEG dialkoxypropyl carbamate, such as ω-methoxy(polyethoxy)ethyl-N-(2,3-di(tetradecanoxy)propyl)carbamate, and 2,3-di(tetradecanoxy)propyl 1-Ν-(ω-methoxy(polyethoxy)ethyl)carbamate.

[0168] In some embodiments, the PEG lipid is PEG2000-DMG:

Chemical formula

[0169] In some embodiments, the PEG lipid is DMG-PEG.

[0170] In some embodiments, the PEG lipid is provided in WO2021 / 026358, WO2017 / 075531, or WO2018 / 081480, each of which is incorporated herein by reference in its entirety.

[0171] In some embodiments, the PEG lipid is a compound of formula II:

Chemical formula

[0172] As described herein, R 8 and R 9 are each independently C 10 -C 30 aliphatic and optionally interrupted by one or more ester bonds. In some embodiments, R 8 and R 9 are each independently C 10 -C 30 aliphatic. In some embodiments, R 8 and R 9 are each independently selected from C 12 -C 16 aliphatic. In some embodiments, R 8 and R 9 are each independently selected from C 12 -C 16 alkyl. In some embodiments, R 8 and R 9 are each independently selected from straight-chain C 12 -C 16 alkyl. In some embodiments, w is an integer from 40 to 50. In some embodiments, w is from 45 to 47. In some embodiments, w is 45.

[0173] In some embodiments, the compound of formula II is 2-[(polyethylene glycol)-2000]-N,N-ditetradecylacetamide (ALC-0159). In some embodiments, the compound of formula II is

Chemical formula

[0174] In some embodiments, the PEG lipid has the following structure:

Chemical formula

[0175] In some embodiments, the PEG lipid is selected from PEG-DAG, PEG-PE, PEG-S-DAG, PEG2000-DMG, PEG-S-DMG, PEG-cer, PEG dialkyloxypropylcarbamate (e.g., ω-methoxy(polyethoxy)ethyl-N-(2,3-di(tetradecanoxy)propyl)carbamate or 2,3-di(tetradecanoxy)propyl-N-(ω-methoxy(polyethoxy)ethyl)carbamate), ALC-0159, and combinations thereof. In some embodiments, the PEG lipid is ALC-0159 or PEG2000-DMG. In some embodiments, the PEG lipid is ALC-0159. In some embodiments, the PEG lipid is PEG2000-DMG. In some embodiments, the PEG lipid is PEG-DAG. In some embodiments, the PEG lipid is PEG-PE. In some embodiments, the PEG lipid is PEG-S-DAG. In some embodiments, the PEG lipid is PEG-cer. In some embodiments, the PEG lipid is PEG dialkyloxypropylcarbamate.

[0176] In some embodiments, the PEG group, which is part of the PEG lipid, has an average number-average molecular weight (M n ) of about 2000 g / mol in a composition comprising one or more PEG lipid molecules.

[0177] In some embodiments, the polymer-conjugated lipid is a polysarcosine-conjugated lipid, also referred to herein as a sarcosylated lipid or pSar lipid. The term "sarcosylated lipid" refers to a molecule that includes both a lipid moiety and a polysarcosine (poly(N-methylglycine) moiety).

[0178] In some embodiments, the polymer-conjugated lipid is a polyoxazoline (POX)-conjugated lipid and / or a polyoxazine (POZ)-conjugated lipid, also referred to herein as an aggregate of a POX and / or POZ polymer and one or more hydrophobic chains, or an oxazolinylated lipid and / or an oxazinylated lipid, or a POX lipid and / or a POZ lipid. The term "oxazolinylated lipid" or "POX lipid" refers to a molecule that includes both a lipid moiety and a polyoxazoline moiety. The term "oxazinylated lipid" or "POZ lipid" refers to a molecule that includes both a lipid moiety and a polyoxazine moiety. The term "oxazolinylated / oxazinylated lipid" or "POX / POZ lipid" or "POXZ lipid" refers to a molecule that includes both a lipid moiety of polyoxazoline and polyoxazine and a portion of the copolymer.

[0179] In some embodiments, the LNPs described herein may include a sarcosylated lipid. In some embodiments, the nucleic acid compositions described herein (e.g., DNA compositions or RNA compositions, particularly mRNA compositions) include a sarcosylated lipid and are substantially free of (or free of) a pegylated lipid.

[0180] In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) comprise the cationic / cationizable lipids described herein and sarcosylated lipids (pSAR-conjugated lipids). In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) may further comprise neutral lipids (e.g., phospholipids, cholesterol or derivatives thereof), or combinations of neutral lipids (e.g., phospholipids and cholesterol or derivatives thereof). In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) comprise the cationic / cationizable lipids, sarcosylated lipids, neutral lipids (e.g., phospholipids), and cholesterol or derivatives thereof described herein. In some embodiments, the phospholipid is DSPC. In some embodiments, the cationic / cationizable lipid is a cationic ionizable lipid of formula (X) (e.g., a cationic ionizable lipid of formula (X-3) or (X-45)). In some embodiments, the cationic / cationizable lipid is a cationic ionizable lipid of formula (XI) (e.g., a cationic ionizable lipid of formula (XIV-1), (XIV-2), or (XIV-3)). In some embodiments, the cationic / cationizable lipid is DPL14, EA-2, or 3D-P-DMA.

[0181] In some embodiments of the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) that include sarcosylated lipids, the composition is substantially free (or free) of pegylated lipids.

[0182] In some embodiments, the sarcosylated lipid comprises from 2 to 200 sarcosine units, such as from 5 to 100 sarcosine units, from 10 to 50 sarcosine units, from 15 to 40 sarcosine units, such as about 23 sarcosine units.

[0183] In some embodiments, the sarcosylated lipid has the structure of the following general formula (XVII): [Chemical formula] and includes wherein s is the number of sarcosine units.

[0184] In some embodiments, the sarcosylated lipid has the structure of the following general formula (XVIII): [Chemical formula] and includes wherein one of 21 R and 22 R contains a hydrophobic group, and the other is H, a hydrophilic group, or a functional group optionally containing a targeting moiety, and x is the number of sarcosine units.

[0185] In some embodiments of formula (XVIII), 21 R is H, a hydrophilic group, or a functional group optionally containing a targeting moiety, and 22 R each contains one or two linear alkyl or alkenyl groups each having at least 12 carbon atoms, for example, at least 14 carbon atoms. In some embodiments, each of the linear alkyl and alkenyl groups has a maximum of 30 carbon atoms, for example, a maximum of 28, maximum of 26, maximum of 24, maximum of 22, maximum of 20, or maximum of 18 carbon atoms. In some embodiments, 22 R each contains one or two linear alkyl or alkenyl groups each having 12 to 30 carbon atoms (for example, 12 to 28 carbon atoms, 12 to 26 carbon atoms, 12 to 24 carbon atoms, 12 to 22 carbon atoms, 12 to 20 carbon atoms, or 12 to 18 carbon atoms).

[0186] In some embodiments, the sarcosylated lipid has the structure of the following general formula (IXX): [Chemical formula] and has In the formula, R is H, a hydrophilic group, or a functional group optionally containing a targeting moiety, and s is the number of sarcosine units.

[0187] In some embodiments, the sarcosylated lipid has the following formula (IXX-1):

Chemical formula

[0188] In some embodiments, the LNPs herein may contain oxazolinylated lipids and / or oxazinylated lipids. In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) contain oxazolinylated lipids and / or oxazinylated lipids and substantially do not contain (or do not contain) pegylated lipids.

[0189] In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) comprise the cationic / cationizable lipids described herein, and oxazolinylated lipids and / or oxazinylated lipids (POX lipids and / or POZ-conjugated lipids). In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) may further comprise neutral lipids (e.g., phospholipids, cholesterol or derivatives thereof), or combinations of neutral lipids (e.g., phospholipids, and cholesterol or derivatives thereof). In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) comprise the cationic / cationizable lipids described herein, oxazolinylated lipids and / or oxazinylated lipids, neutral lipids (e.g., phospholipids), and cholesterol or derivatives thereof. In some embodiments, the phospholipid is DSPC. In some embodiments, the cationic / cationizable lipid is a cationic ionizable lipid of formula (X) (e.g., a cationic ionizable lipid of formula (X-3) or (X-45)). In some embodiments, the cationic / cationizable lipid is a cationic ionizable lipid of formula (XI) (e.g., a cationic ionizable lipid of formula (XIV-1), (XIV-2), or (XIV-3)). In some embodiments, the cationic / cationizable lipid is DPL14, EA-2, or 3D-P-DMA. In some embodiments of the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) that comprise oxazolinylated and / or oxazinylated lipids, the composition is substantially free (or free) of pegylated lipids.

[0190] In some embodiments, in the oxazolinated and / or oxazinized lipid (i.e., (i) a POX and / or POZ polymer and (ii) a conjugate containing one or more hydrophobic chains), the components (i) and (ii) are linked to each other via a linker containing at least one functional moiety. In some embodiments, the linker comprises an alkylene moiety substituted with at least one monovalent functional moiety. In some embodiments, the linker comprises an alkylene group and a divalent functional moiety, the divalent functional moiety linking the alkylene group to one or more hydrophobic chains, and the alkylene group binding to the POX and / or POZ polymer. In some embodiments, the linker comprises an alkylene group and a divalent functional moiety, the divalent functional moiety linking the alkylene group to one or more hydrophobic chains, the alkylene group being substituted with at least one monovalent functional moiety, and the alkylene group binding to the POX and / or POZ polymer.

[0191] In some embodiments of the oxazolinated and / or oxazinized lipid, each monovalent functional moiety is independently selected from hydroxy, ether, halogen, cyano, azide, nitro, amino, ammonium, ester, carboxyl, thiol (sulfanyl), disulfanyl, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamido, sulfonamido, sulfamate, diselenide, dithionamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imidate, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbodithioate, carbonotrithioate, guanidino (imidamide), carbamimidate, carbodiimidate, carbamate, carbamodithioate, carbodithioimidate, carbamimidothioate, carbamothioate, carbodiimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imide, and amide moieties.

[0192] In some embodiments of the oxazolinylated and / or oxazinylated lipids, each divalent functional moiety is independently an ether, amino, ester, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamido, sulfonamido, sulfamate, diselenide, dithioamide, disulfate diamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imidate, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbodithioate, carbonotrithioate, guanidino (imidamide), carbamimidate, carbodiimidate, carbamate, carbamodithioate, carbodithioimidate, carbamimidothioate, carbamothioate, carbodiimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imine, imide, and amide moiety.

[0193] In some embodiments, the oxazolinylated and / or oxazinylated lipids have the following structure (in particular, when one or more hydrophobic chains are attached to the N-terminus (i.e., the terminal N atom) of the POX and / or POZ polymer, as shown, for example, in Formula (XXI) herein): (Hydrophobic chain) 1-2 -(An alkylene moiety substituted with at least one monovalent functional moiety)-(POX and / or POZ polymer), [(Hydrophobic chain)-(Divalent functional moiety)] 1-2 -One of (an alkylene moiety)-(POX and / or POZ polymer).

[0194] In some embodiments, the oxazolinylated and / or oxazinylated lipids have the following formula (in particular, when one or more hydrophobic chains are attached to the N-terminus (i.e., the terminal N atom) of the POX and / or POZ polymer, as shown, for example, in Formula (XXI) herein): (Hydrophobic chain) 1-2-(An alkylene moiety substituted with at least one monovalent functional moiety)-(a POX and / or POZ polymer)-(a terminal group), [(A hydrophobic chain)-(a divalent functional moiety)] 1-2 -Has one of -(an alkylene moiety)-(a POX and / or POZ polymer)-(a terminal group).

[0195] In some embodiments of the oxazolinylated and / or oxazinylated lipids, the alkylene moiety substituted with at least one monovalent functional moiety is substituted with one or more (e.g., from 1 to the maximum number of hydrogen atoms bonded to the alkylene moiety, e.g., 1, 2, 3, 4, 5, or 6, e.g., 1 - 5, 1 - 4, or 1 - 3, or 1 or 2) independently selected monovalent functional moieties.

[0196] In some embodiments of the oxazolinylated and / or oxazinylated lipids, the alkylene moiety is C 1-6 -alkylene, e.g., C 1-3 -alkylene, e.g., methylene, ethylene, or trimethylene.

[0197] In some embodiments (particularly where one or more hydrophobic chains are attached to the C-terminus (i.e., the terminal C atom) of the POX and / or POZ polymer, e.g., as shown in formula (XXI’) herein), the linker contains at least one bifunctional moiety through which one or more hydrophobic chains are attached to the POX and / or POZ polymer. In some embodiments, the linker is an alkylene moiety (e.g., C 1-6 alkylene moiety, e.g., C 1-3 alkylene moiety), a cycloalkylene moiety (preferably C 3-8 -cycloalkylene, e.g., C 3-6 -cycloalkylene moiety), or a cycloalkenylene moiety (preferably C 3-8 -cycloalkenylene, e.g., C 3-6- cycloalkenylene moiety), and each of these may further include a bifunctional moiety that connects to the POX and / or POZ polymer (either directly to the end of the POX and / or POZ polymer or preferably through an additional bifunctional moiety). For example, one hydrophobic chain may be attached to the end of the POX and / or POZ polymer through one bifunctional moiety (either directly, or through an alkylene, cycloalkylene, or cycloalkenylene moiety, or through an alkylene, cycloalkylene, or cycloalkenylene moiety having another bifunctional moiety), or two hydrophobic chains may be attached to the end of the POX and / or POZ polymer through two bifunctional moieties (which preferably attaches to an alkylene, cycloalkylene, or cycloalkenylene moiety or to an alkylene, cycloalkylene, or cycloalkenylene moiety having another bifunctional moiety), or two hydrophobic chains may be attached to the end of the POX and / or POZ polymer through the same bifunctional moiety (which is a trifunctional moiety and can be attached either directly to the end of the POX and / or POZ polymer or through an alkylene, cycloalkylene, or cycloalkenylene moiety or through an alkylene, cycloalkylene, or cycloalkenylene moiety having another bifunctional moiety).In some embodiments, each difunctional moiety is independently selected from ether, amino, ester, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamide, sulfonamide, sulfamate, diselenide, dithioamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imidate, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbodithioate, carbonotrithioate, guanidino (imidamide), carbamimidate, carbodiimidate, carbamate, carbamodithioate, carbodithioimidate, carbamimidothioate, carbamothioate, carbodiimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imine, imide, and amide moieties.

[0198] In some embodiments of oxazolinylated and / or oxazinylated lipids, the cycloalkylene moiety is C 3-8 -cycloalkylene, e.g., C 3-6 -cycloalkylene, e.g., cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, and the cycloalkylene moiety is optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents (e.g., independently selected from the group consisting of OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl).

[0199] In some embodiments of oxazolinylated and / or oxazinylated lipids, the cycloalkenylene moiety is C 3-8 -cycloalkenylene, e.g., C 3-6 -cycloalkenylene, e.g., cyclopropenylene, cyclobutenylene, cyclopentenylene, cyclohexenylene, and the cycloalkenylene moiety is optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents (e.g., independently selected from the group consisting of OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3optionally substituted with one selected from the group consisting of -alkyl).

[0200] In some embodiments of the oxazolinated and / or oxazinized lipids, the alkylene moiety is C 1-6 -alkylene, e.g., C 1-3 -alkylene, e.g., methylene, ethylene, or trimethylene, or C 2-3 alkylene.

[0201] In some embodiments (especially those in which one or more hydrophobic chains are attached to the C-terminus (i.e., the terminal C atom) of a POX and / or POZ polymer, as shown, for example, in formula (XXI') below), the oxazolinated and / or oxazinized lipids have the following structure: (hydrophobic chain)-(divalent functional moiety)-(POX and / or POZ polymer) [(hydrophobic chain)-(divalent functional moiety)] 1-2 -(alkylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer) (hydrophobic chain)-(divalent functional moiety)-(cycloalkylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer) (hydrophobic chain)-(divalent functional moiety)-(cycloalkenylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer) (hydrophobic chain)-(divalent functional moiety)-(alkylene moiety)-(POX and / or POZ polymer) [(hydrophobic chain 2 -(trivalent functional site)]-(alkylene site)-(divalent functional moiety)-(POX and / or POZ polymer) and may have the general formula (XXI').

[0202] In some embodiments (especially those in which one or more hydrophobic chains are attached to the C-terminus (i.e., the terminal C atom) of a POX and / or POZ polymer, as shown, for example, in formula (XXI') below), the oxazolinated and / or oxazinized lipids have the following formula: (Hydrophobic chain)-(Divalent functional moiety)-(POX and / or POZ polymer)-(Terminal group) [(Hydrophobic chain)-(Divalent functional moiety)] 1-2 -(Alkylene moiety)-(Divalent functional moiety)-(POX and / or POZ polymer)-(Terminal group) (Hydrophobic chain)-(Divalent functional moiety)-(Cycloalkylene moiety)-(Divalent functional moiety)-(POX and / or POZ polymer)-(Terminal group) (Hydrophobic chain)-(Divalent functional moiety)-(Cycloalkenylene moiety)-(Divalent functional moiety)-(POX and / or POZ polymer)-(Terminal group) (Hydrophobic chain)-(Divalent functional moiety)-(Alkylene moiety)-(POX and / or POZ polymer)-(Terminal group) [(Hydrophobic chain) 2 -(Trivalent functional site)]-(Alkylene site)-(Divalent functional moiety)-(POX and / or POZ polymer)-(Terminal group), and one of them (and may correspond to the general formula (XXI’)).

[0203] The POX and / or POZ polymer may contain a neutral terminal group (e.g., H, alkyl, alkoxy, ester, or amide terminal group) or a functionalized terminal group (e.g., hydroxy, thiol, cyano, azide, or amino terminal group). In the case of nucleic acid-lipid particles, the POX and / or POZ polymer is conjugated, preferably covalently, with one or more hydrophobic chains.

[0204] In certain embodiments of oxazolinated and / or oxazinized lipids, the terminal group of the POX and / or POZ polymer can be functionalized with one or more molecular moieties that impart specific properties, such as a positive or negative charge, or targeting agents that would direct the particle to a specific cell type, collection of cells, or tissue.

[0205] A variety of suitable targeting agents are known in the art. Non-limiting examples of targeting agents include peptides, proteins, enzymes, nucleic acids, fatty acids, hormones, antibodies, carbohydrates, monosaccharides, oligosaccharides, or polysaccharides, peptidoglycans, glycopeptides, and the like. In some embodiments, the targeting agent comprises a targeting pair, such as the following pairs: antigen - an antibody specific for the antigen; avidin - streptavidin; folate - folate receptor; transferrin - transferrin receptor; aptamer - a molecule to which the aptamer is specific (e.g., pegaptanib - VEGF receptor); arginine - glycine - aspartic acid (RGD) peptide - α v β 3 integrin; asparagine - glycine - arginine (NGR) peptide - aminopeptidase N; galactose - asialoglycoprotein receptor. For example, any of several different substances that bind to an antigen on the surface of a target cell can be used. Antibodies against target cell surface antigens will generally exhibit the specificity required for targeting. In addition to antibodies, suitable immunoreactive fragments, such as Fab, Fab′, F(ab′)2, or scFv fragments, or single - domain antibodies (e.g., camelid V H H fragments) can also be used. Many antibody fragments suitable for use in forming the targeting mechanism are already available in the art. Similarly, a ligand for any receptor on the surface of a target cell can be suitably used as a targeting agent. These include any natural or synthetic small molecule or biomolecule that specifically binds to a cell surface receptor, protein, or glycoprotein found on the surface of the desired target cell.

[0206] In certain embodiments of the oxazolinated and / or oxazinized lipid, the POX and / or POZ polymer comprises 2 to 200, 2 to 190, 2 to 180, 2 to 170, 2 to 160, 2 to 150, 2 to 140, 2 to 130, 2 to 120, 2 to 110, 2 to 100, 2 to 90, 2 to 80, 2 to 70, 5 to 200, 5 to 190, 5 to 180, 5 to 170, 5 to 160, 5 to 150, 5 to 140, 5 to 130, 5 to 120, 5 to 110, 5 to 100, 5 to 90, 5 to 80, 5 to 70, 10 to 200, 10 to 190, 10 to 180, 10 to 170, 10 to 160, 10 to 150, 10 to 140, 10 to 130, 10 to 120, 10 to 110, 10 to 100, 10 to 90, 10 to 80, or 10 to 70 POX and / or POZ repeating units.

[0207] In some embodiments, the POX and / or POZ polymer in the oxazolinated and / or oxazinized lipid has the following general formula (XX):

Chemical formula

[0208] In some embodiments of the oxazolinated and / or oxazinized lipid, the POX and / or POZ polymer is a polymer of POX and has the following general formula (XXa):

Chemical formula

[0209] In some embodiments of the oxazolinated and / or oxazinized lipid, the POX and / or POZ polymer is a polymer of POZ and has the following general formula (XXb):

Chemical formula

[0210] In any of the above embodiments of formula (XX), (XXa), and (XXb), m (i.e., the number of repeating units of formula (XXa) or formula (XXb) in the polymer) is preferably 2 to 190, for example, 2 to 180, 2 to 170, 2 to 160, 2 to 150, 2 to 140, 2 to 130, 2 to 120, 2 to 110, 2 to 100, 2 to 90, 2 to 80, 2 to 70, 5 to 200, 5 to 190, 5 to 180, 5 to 170, 5 to 160, 5 to 150, 5 to 140, 5 to 130, 5 to 120, 5 to 110, 5 to 100, 5 to 90, 5 to 80, 5 to 70, 10 to 200, 10 to 190, 10 to 180, 10 to 170, 10 to 160, 10 to 150, 10 to 140, 10 to 130, 10 to 120, 10 to 110, 10 to 100, 10 to 90, 10 to 80, or 10 to 70. In a specific embodiment of any of the above embodiments of formula (XX), (XXa), and (XXb), m is 2 to 180, for example, 4 to 160, 6 to 140, 8 to 120, or 10 to 100, for example, 20 to 80, 30 to 70, or 40 to 50.

[0211] In some embodiments of oxazolinated and / or oxazinized lipids, the POX and / or POZ polymers are the following general formulas (XXa) and (XXb):

Chemical formula

[0212] In some embodiments of the oxazolinated and / or oxazinized lipids, the number of repeating units of formula (XXa) in the copolymer is from 1 to 179, such as from 1 to 159, from 1 to 139, from 1 to 119, or from 1 to 99, and the number of repeating units of formula (XXb) in the copolymer is from 1 to 179, such as from 1 to 159, from 1 to 139, from 1 to 119, or from 1 to 99, and the sum of the number of repeating units of formula (XXa) and the number of repeating units of formula (XXb) in the copolymer is from 2 to 180, such as from 4 to 160, from 6 to 140, from 8 to 120, or from 10 to 100, such as from 20 to 80, from 30 to 70, or from 40 to 50.

[0213] In some of the above embodiments of formula (XX), (XXa), and (XXb), R at each occurrence (i.e., in each repeating unit) 11 can be the same alkyl group (e.g., R 11 can be methyl in each repeating unit). In some alternative embodiments of formula (XX), (XXa), and (XXb), R 11 in at least one repeating unit is different from R 11 in another repeating unit (e.g., for at least one repeating unit, R 11 is one particular alkyl (e.g., ethyl), and for at least one different repeating unit, R 11 is a different particular alkyl (e.g., methyl)). For example, each R 11 can be selected from two different alkyl groups (e.g., methyl and ethyl), and not all R 11 are the same alkyl.

[0214] In any of the above embodiments of formula (XX), (XXa), and (XXb), R 11 is preferably methyl or ethyl, more preferably methyl. Thus, in some embodiments of formula (XX), (XXa), and (XXb), each R 11 is methyl, or each R 11is ethyl. In some alternative embodiments of formula (XX), (XXa), and (XXb), R 11 is independently selected from methyl and ethyl for each repeating unit, and in at least one repeating unit, R 11 is methyl and in at least one repeating unit R 11 is ethyl.

[0215] In some embodiments, the oxazolinated and / or oxazinylated lipid has the following general formula (XXI) or (XXI’):

Chemical formula

[0216] In some embodiments (particularly, oxazolinylated and / or oxazinylated lipids of formulas (XXI) and (XXI’)), the targeting pair is selected from the following pairs: antigen - antibody specific for said antigen; avidin - streptavidin; folate - folate receptor; transferrin - transferrin receptor; aptamer - molecule to which said aptamer is specific; arginine - glycine - aspartic acid (RGD) peptide - α v β 3 integrin; asparagine - glycine - arginine (NGR) peptide - aminopeptidase N; galactose - asialoglycoprotein receptor. Thus, in some embodiments, a member of the targeting pair comprises one of an antigen, antibody, avidin, streptavidin, folate, transferrin, aptamer, RGD peptide, NGR peptide, and galactose.

[0217] In some embodiments of formula (XXI), a is 1, i.e., the oxazolinylated and / or oxazinylated lipid has the following general formula (XXIa) or (XXIa’):

Chemical formula

[0218] In some embodiments of formula (XXI), a is 2, i.e., the oxazolinated and / or oxazinylated lipid is of the following general formula (XXIb) or (XXIb’):

Chemical formula

[0219] In any of the above embodiments of formulae (XXIa), (XXIa’), (XXIb), and (XXIb’), R 11 , R 12 , R 13 , and m are as defined in formula (XXI) / (XXI’).

[0220] In some of the above embodiments of formulae (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), and (XXIb’), R 11 at each occurrence (i.e., in each repeating unit) can be the same alkyl group (e.g., R 11 can be methyl in each repeating unit). In some alternative embodiments of formulae (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), and (XXIb’), R 11 in at least one repeating unit is different from R 11 in another repeating unit (e.g., for at least one repeating unit, R 11 is a particular alkyl (e.g., ethyl), and for at least one different repeating unit, R 11 is a different particular alkyl (e.g., methyl)). For example, each R 11 can be selected from two different alkyl groups (e.g., methyl and ethyl), and not all R 11 are the same alkyl.

[0221] In any of the above embodiments of formulae (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), and (XXIb’), R11 is preferably methyl or ethyl, more preferably methyl. Thus, in some embodiments of formulae (XXI), (XXI'), (XXIa), (XXIa'), (XXIb), and (XXIb'), each R 11 is methyl, or each R 11 is ethyl. In some alternative embodiments of formulae (XXI), (XXI'), (XXIa), (XXIa'), (XXIb), and (XXIb'), R 11 is independently selected from methyl and ethyl for each repeating unit, and in at least one repeating unit, R 11 is methyl and in at least one repeating unit R 11 is ethyl.

[0222] In any of the above embodiments of formulae (XXI), (XXI'), (XXIa), (XXIa'), (XXIb), and (XXIb'), m is preferably preferably 2 to 190, for example, 2 to 180, 2 to 170, 2 to 160, 2 to 150, 2 to 140, 2 to 130, 2 to 120, 2 to 110, 2 to 100, 2 to 90, 2 to 80, 2 to 70, 5 to 200, 5 to 190, 5 to 180, 5 to 170, 5 to 160, 5 to 150, 5 to 140, 5 to 130, 5 to 120, 5 to 110, 5 to 100, 5 to 90, 5 to 80, 5 to 70, 10 to 200, 10 to 190, 10 to 180, 10 to 170, 10 to 160, 10 to 150, 10 to 140, 10 to 130, 10 to 120, 10 to 110, 10 to 100, 10 to 90, 10 to 80, or 10 to 70. In certain embodiments of formulae (XXI), (XXI'), (XXIa), (XXIa'), (XXIb), and (XXIb'), m is 2 to 180, for example, 4 to 160, 6 to 140, 8 to 120, or 10 to 100, for example, 20 to 80, 30 to 70, or 40 to 50.

[0223] In some embodiments of formulae (XXI), (XXI'), (XXIIa), (XXIa'), (XXIb), and (XXIb'), L 11is an alkylene moiety substituted with at least one functional moiety, e.g., at least one monovalent functional moiety, and / or an alkylene moiety bonded to a divalent functional moiety at a terminal where the alkylene group is R 14 and includes an alkylene moiety bonded to a divalent functional moiety at the terminal where the alkylene group is bonded to R. Preferably, each monovalent functional moiety is independently selected from hydroxy, ether, halogen, cyano, azide, nitro, amino, ammonium, ester, carboxyl, thiol (sulfanyl), disulfanyl, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamide, sulfonamide, sulfamate, diselenide, dithionamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbondithioate, carbonotrithioate, guanidino (imidamide), carbamimidate, carbonimidate, carbamate, carbamodithioate, carbondithioimidate, carbamimidothioate, carbamothioate, carbonimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imide, and amide, and / or each divalent functional moiety is independently selected from ether, amino, ester, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamide, sulfonamide, sulfamate, diselenide, dithionamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imidate, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbondithioate, carbonotrithioate, guanidino (imidamide), carbamimidate, carboimidate, carbamate, carbamodithioate, carbondithioimidate, carbamimidothioate, carbamothioate, carboimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imine, imide, and amide.

[0224] In some embodiments of Formulas (XXI), (XXI'), (XXIa), (XXIa'), (XXIb), and (XXIb'), L 11 comprises an alkylene moiety substituted with at least one monovalent functional moiety as specified above. Thus, in some embodiments, the oxazolinylated and / or oxazinylated lipid has the following structure (in particular, when one or more hydrophobic chains are attached to the N-terminus (i.e., the terminal N atom) of the POX and / or POZ polymer, as shown, for example, in Formula (XXI)): (Hydrophobic chain) 1-2 - (an alkylene moiety substituted with at least one monovalent functional moiety) - (POX and / or POZ polymer) and may include, wherein "hydrophobic chain" represents R 14 "an alkylene moiety substituted with at least one monovalent functional moiety" represents L 11 and "POX and / or POZ polymer" represents the polymer specified in Formula (XX).

[0225] In some embodiments, the oxazolinylated and / or oxazinylated lipid has the following Formula (XXIc) (in particular, when one or more hydrophobic chains are attached to the N-terminus (i.e., the terminal N atom) of the POX and / or POZ polymer, as shown, for example, in Formula (XXI)): (Hydrophobic chain) 1-2 - (an alkylene moiety substituted with at least one monovalent functional moiety) - (POX and / or POZ polymer) - R 13 and has.

[0226] In some embodiments (particularly of the oxazolinylated and / or oxazinylated lipids of formula (XXIc)), at least one monovalent functional moiety can be any one of the monovalent functional moieties specified herein, for example, hydroxy, ether, halogen, cyano, azide, nitro, amino, ammonium, ester, carboxyl, thiol (sulfanyl), disulfanyl, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamido, sulfonamido, sulfamate, diselenide, dithioamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbonodithioate, carbonotritioate, guanidino (imidamide), carbamimidate, carbonimidate, carbamate, carbamodithioate, carbonodithioimidate, carbamimidothioate, carbamothioate, carbonimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imide, and amide, selected from the group consisting of.

[0227] In some embodiments (particularly of the oxazolinylated and / or oxazinylated lipids of formula (XXIc)), the alkylene moiety substituted with at least one monovalent functional moiety is C 1-6 alkylene, for example, C 1-3 alkylene, for example, methylene, ethylene, or trimethylene.

[0228] In some embodiments (particularly of the oxazolinylated and / or oxazinylated lipids of formula (XXIc)), the alkylene moiety substituted with at least one monovalent functional moiety is substituted with one or more (for example, from 1 to the maximum number of hydrogen atoms bonded to the alkylene moiety, for example, 1, 2, 3, 4, 5, or 6, for example, 1 - 5, 1 - 4, or 1 - 3, or 1 or 2) independently selected monovalent functional moieties.

[0229] In some embodiments (particularly those of the oxazolinated and / or oxazinylated lipids of formula (XXIc)), the alkylene moiety substituted with at least one monovalent functional moiety is C 1-6 -alkylene, e.g., C 1-3 -alkylene, e.g., methylene, ethylene, or trimethylene, and is substituted with one or more (e.g., from 1 up to the maximum number of hydrogen atoms bonded to the alkylene moiety, e.g., 1, 2, 3, 4, 5, or 6, e.g., 1-5, 1-4, or 1-3, or 1 or 2) independently selected monovalent functional moieties.

[0230] In some embodiments of formulas (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), and (XXIb’), L 11 is an alkylene moiety that at the terminus where the alkylene group is bonded to R 14 is bonded to a divalent functional moiety as specified above. Thus, in some embodiments, the oxazolinated and / or oxazinylated lipid has the following structure (particularly when one or more hydrophobic chains are bonded to the N-terminus (i.e., the terminal N atom) of the POX and / or POZ polymer, e.g., as shown in formula (XXI)): [(hydrophobic chain)-(divalent functional moiety)] 1-2 -(alkylene moiety)-(POX and / or POZ polymer), and may include wherein "hydrophobic chain" represents R 14 and "-(divalent functional moiety) 1-2 -(alkylene moiety)" represents L 11 and "POX and / or POZ polymer" represents the polymer specified in formula (XX).

[0231] In some embodiments, the oxazolinated and / or oxazinylated lipid has the following formula (XXId) (particularly when one or more hydrophobic chains are bonded to the N-terminus (i.e., the terminal N atom) of the POX and / or POZ polymer, e.g., as shown in formula (XXI)): [(hydrophobic chain)-(divalent functional moiety)] 1-2-(alkylene moiety)-(POX and / or POZ polymer)-R 13 has.

[0232] In some embodiments (particularly of the oxazolinated and / or oxazinized lipids of formula (XXId)), the divalent functional moiety can be any one of the divalent functional moieties specified herein, for example, ether, amino, ester, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamido, sulfonamido, sulfamate, diselenide, dithioamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imidate, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbonodithioate, carbonotritioate, guanidino (imidamide), carbamimidate, carbodiimidate, carbamate, carbamodithioate, carbonodithioimidate, carbamimidothioate, carbamothioate, carbodiimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imine, imide, and amide. In some embodiments, the linker comprises at least one divalent functional moiety selected from the group consisting of ester, sulfide, disulfide, sulfone, orthoester, acylhydrazone, hydrazine, oxime, acetal, ketal, amino, and amide moieties. In some preferred embodiments, the linker comprises at least one divalent functional moiety selected from the group consisting of ester, sulfide, sulfone, amino, and amide moieties.

[0233] In some embodiments (particularly of the oxazolinated and / or oxazinized lipids of formula (XXId)), the alkylene moiety is C 1-6 -alkylene, for example, C 1-3 -alkylene, for example, methylene, ethylene, or trimethylene.

[0234] In some embodiments of Formulas (XXI), (XXI'), (XXIa), (XXIa'), (XXIb), (XXIb'), (XXIc), and (XXId) (in particular, where one or more hydrophobic chains are attached to the N-terminus (i.e., the terminal N atom) of the POX and / or POZ polymer, as shown, for example, in Formula (XXI)), L 11 comprises at least one ester, sulfide, disulfide, sulfone, orthoester, acylhydrazone, hydrazine, oxime, acetal, ketal, or amide moiety. In certain embodiments, L 11 is [*-NHC(O)] p (C 1-6 -alkylene), [*-C(O)NH] p (C 1-6 -alkylene)-, [*-C(O)O] p (C 1-6 -alkylene)-, [*-OC(O)] p -(C 1-6 -alkylene)-, [*-S] p (C 1-6 -alkylene)-, [*-SS] p (C 1-6 -alkylene)-, [*-S(O) 2 p (C 1-6 -alkylene)-, [(*-O) r C(OR 25 ) 3-r (C 1-6 -alkylene)-, [*-C(OR 25 ) 2 O] p (C 1-6 -alkylene)-, [*-C(R 25 )(=N-N(R 26 )C(O)-)] p (C 1-6 -alkylene)-, [*-C(O)(N(R 26 )-N=)C(R 25 )-] p (C 1-6 -alkylene)-, [*=C(=N-N(R 26 )C(O)(R 25 ))] p (C 1-6 ​-(alkylene)-, [*N(R 26 )N(R 26 )] p (C 1-6 -alkylene)-, [*=C(=N(OH))] p (C 1-6 -alkylene)-, and [*-OC(R 25 )(R 26 )O] p (C 1-6 -alkylene)-selected from the group consisting of, * represents the point of attachment to R 14 , p is 1 or 2, C 1-6 -alkylene is either divalent (when p is 1) or trivalent (when p is 2), R 25 is selected from the group consisting of C 1-6 alkyl, aryl, and aryl-(C 1-6 alkyl), R 26 is H, C 1-6 alkyl, aryl, and aryl(C 1-6 alkyl), r is an integer from 1 to 2. For example, L 11 is [*-NHC(O)] p (C 1-3 -alkylene)-, [*-C(O)NH] p (C 1-3 -alkylene)-, [*-C(O)O] p (C 1-3 -alkylene)-, [*-OC(O)] p (C 1-3 -alkylene)-, [*-S] p (C 1-3 -alkylene)-, [*-SS] p (C 1-3 -alkylene)-, [*-S(O) 2 p (C 1-3 -alkylene)-, [(*-O) r C(OR 25 ) 3-r (C 1-3 -alkylene)-, [*-C(OR 25 ) 2 O] p (C 1-3 -alkylene)-, [*-C(R​25 )(=N-N(R 26 )C(O)-)] p (C 1-3 -alkylene)-, [*-C(O)(N(R 26 )-N=)C(R 25 )-] p (C 1-3 -alkylene)-, [*=C(=N-N(R 26 )C(O)(R 25 ))] p (C 1-3 -alkylene)-, [*N(R 26 )N(R 26 )] p (C 1-3 -alkylene)-, [*=C(=N(OH))] p (C 1-3 -alkylene)-, and [*-OC(R 25 )(R 26 )O] p (C 1-3 -alkylene)- may be selected from the group consisting of, * represents the point of attachment to R 14 , p is 1 or 2, C 1-3 -alkylene is either divalent (when p is 1) or trivalent (when p is 2), R 25 is, C 1-6 alkyl, aryl, and aryl(C 1-6 alkyl) selected from the group consisting of, R 26 is, H, C 1-6 alkyl, aryl, and aryl(C 1-6 alkyl) selected from the group consisting of, r is an integer from 1 to 2.

[0235] In some embodiments, R 25 is, C 1-3 alkyl, phenyl, and phenyl(C 1-3 alkyl) selected from the group consisting of, for example, methyl, ethyl, phenyl, benzyl, and phenylethyl selected from the group consisting of.

[0236] In some embodiments, R 26 is, H, C 1-3 alkyl, phenyl, and phenyl(C13 It is selected from the group consisting of, for example, H, methyl, ethyl, phenyl, benzyl, and phenylethyl, from the group consisting of (alkyl).

[0237] In some embodiments of Formulas (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), (XXIb’), (XXIc), and (XXId) (in particular, where one or more hydrophobic chains are, for example, as shown in Formula (XXI), attached to the N-terminus (i.e., the terminal N atom) of the POX and / or POZ polymer), L 11 is [*-NHC(O)] p (C 1-6 -alkylene)-, [*-C(O)NH] p (C 1-6 -alkylene)-, [*-C(O)O] p (C 1-6 -alkylene)-, [*-OC(O)] p (C 1-6 -alkylene)-, [*-S] p (C 1-6 -alkylene)-, and [*-S(O) 2 p (C 1-6 -alkylene)-, preferably from the group consisting of [*-NHC(O)] p (C 1-6 -alkylene)-, [*-C(O)O] p (C 1-6 -alkylene)-, [*-OC(O)] p (C 1-6 -alkylene)-, [*-S] p (C 1-6 -alkylene)-, and [*-S(O) 2 p (C 1-6 -alkylene)-, more preferably selected from the group consisting of [*-NHC(O)] p (C 1-6 -alkylene)-, and [*-C(O)O] p (C 1-6 -alkylene)-, where * represents the point of attachment to R 14 and p is 1 or 2, C 1-6 ​​-alkylene is either divalent (when p is 1) or trivalent (when p is 2).

[0238] In some embodiments of formulas (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), (XXIb’), (XXIc), and (XXId) (in particular, where one or more hydrophobic chains are attached to the N-terminus (i.e., the terminal N atom) of a POX and / or POZ polymer, as shown, for example, in formula (XXI)), L 11 is [*-NHC(O)] p (C 1-3 -alkylene)-, [*-C(O)NH] p (C 1-3 -alkylene)-, [*-C(O)O] p (C 1-3 -alkylene)-, [*-OC(O)] p -(C 1-3 -alkylene)-, [*-S] p (C 1-3 -alkylene)-, and [*-S(O) 2 p (C 1-3 -alkylene)-, preferably selected from the group consisting of [*-NHC(O)] p (C 1-3 -alkylene)-, [*-C(O)O] p (C 1-3 -alkylene)-, [*-OC(O)] p (C 1-3 -alkylene)-, [*-S] p (C 1-3 -alkylene)-, and [*-S(O) 2 p (C 1-3 -alkylene)-, more preferably selected from the group consisting of [*-NHC(O)] p (C 1-3 -alkylene)-, and [*-C(O)O] p (C 1-3 -alkylene)-, where * represents the point of attachment to R 14 , p is 1 or 2, and C 1-3 ​​- The alkylene is either divalent (when p is 1) or trivalent (when p is 2).

[0239] In some embodiments of Formulas (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), (XXIb’), (XXIc), and (XXId) (particularly, where one or more hydrophobic chains are attached to the N-terminus (i.e., the terminal N atom) of a POX and / or POZ polymer, as shown, for example, in Formula (XXI)), L 11 is selected from the group consisting of *-NHC(O)-(CH 2 )-, *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 )-, *-C(O)NH-(CH 2 ) 2 -, -(CH 2 )-CH(OC(O)-*)-(CH 2 OC(O)-*), -(CH 2 )-CH(S-*) 2 -, -(CH 2 )-CH(S-*)-CH 2 (S-*), *-S-(CH 2 ) 3 -, *-S(O) 2 -(CH 2 ) 3 -, and *-OC(O)-(CH 2 )-, where * represents the point of attachment to R 14 . Thus, R 12 is selected from the group consisting of R 14 , -L 11 R 14 , -(CH 2 )-CH(OC(O)R 14 )(CH 2 OC(O)R 14 ), -(CH 2 )-CH(SR 14 ) 2 , and -(CH 2 )-CH(SR 14 )-CH 2 (SR 14 ), and L 11 is *-NHC(O)-(CH2 )-, *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 )-, *-C(O)NH-(CH 2 ) 2 -, *-S-(CH 2 ) 3 -, *-S(O) 2 -(CH 2 ) 3 -, and *-OC(O)-(CH 2 )- selected from the group consisting of, preferably, L 11 is, *-NHC(O)-(CH 2 )- or *-NHC(O)-(CH 2 ) 2 -, where * represents the point of attachment to R 14 .

[0240] In some embodiments of Formulas (XXI), (XXIa), and (XXIb), R 12 is, -L 11 (R 14 ) p , i.e., the POX and / or POZ polymer is linked to one or more hydrophobic chains (i.e., R 11 ) via a linker L 14 .

[0241] In some embodiments of Formulas (XXI’), (XXIa’), and (XXIb’), the linker contains at least one bifunctional moiety through which one or more hydrophobic chains (R 14 ) are attached to the C-terminus of the POX and / or POZ polymer. In some embodiments, the linker is an alkylene moiety (e.g., a C 1-6 alkylene moiety, e.g., a C 1-3 alkylene moiety), a cycloalkylene moiety (preferably a C 3-8 -cycloalkylene, e.g., a C 3-6 -cycloalkylene moiety), or a cycloalkenylene moiety (preferably a C 3-8 -cycloalkenylene, e.g., a C 3-6- cycloalkenylene moiety), and each of these may further comprise a bifunctional moiety connecting the C-terminal POX and / or POZ polymer (either directly to the C-terminus or preferably via an additional bifunctional moiety). For example, one hydrophobic chain (R 14 ) may be attached to the C-terminus of the POX and / or POZ polymer via one bifunctional moiety (either directly or via an alkylene, cycloalkylene, or cycloalkenylene moiety, or via an alkylene, cycloalkylene, or cycloalkenylene moiety having another bifunctional moiety), or two hydrophobic chains (R 14 ) may be attached to the C-terminus of the POX and / or POZ polymer via two bifunctional moieties (which preferably attach to an alkylene, cycloalkylene, or cycloalkenylene moiety or an alkylene, cycloalkylene, or cycloalkenylene moiety having another bifunctional moiety), or two hydrophobic chains (R 14) can be attached to the C-terminus of the POX and / or POZ polymer via the same bifunctional moiety (which is a trifunctional moiety and can be attached directly to the C-terminus of the POX and / or POZ polymer or via an alkylene, cycloalkylene, or cycloalkenylene moiety or via an alkylene, cycloalkylene, or cycloalkenylene moiety having another bifunctional moiety). In some embodiments, each divalent functional moiety is independently selected from the group consisting of ether, amino, ester, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamide, sulfonamide, sulfamate, diselenide, dithioamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imidate, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbodithioate, carbonotrithioate, guanidino (imidamide), carbamimidate, carbodiimidate, carbamate, carbamodithioate, carbodithioimidate, carbamimidothioate, carbamothioate, carbodiimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imine, imide, and amide moieties. In some preferred embodiments of Formulas (XXI'), (XXIa'), and (XXIb'), the linker comprises at least one divalent functional moiety selected from the group consisting of amide, sulfide, sulfone, and amino moieties.

[0242] In some embodiments of Formulas (XXI'), (XXIa'), and (XXIb'), the cycloalkylene moiety is C 3-8 -cycloalkylene, for example, C 3-6 -cycloalkylene, for example, cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, and the cycloalkylene moiety is optionally substituted, for example, with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl.

[0243] In some embodiments of formulas (XXI'), (XXIa'), and (XXIb'), the cycloalkenylene moiety is C 3-8 -cycloalkenylene, for example, C 3-6 -cycloalkenylene, for example, cyclopropenylene, cyclobutenylene, cyclopentenylene, cyclohexenylene, and the cycloalkenylene moiety is optionally substituted, for example, with one or more (e.g., 1, 2, 3, or 4) substituents (e.g., independently, -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl).

[0244] In some embodiments of formulas (XXI'), (XXIa'), and (XXIb'), the alkylene moiety is C 1-6 -alkylene, for example, C 1-3 -alkylene, for example, methylene, ethylene, or trimethylene, or C 2-3 alkylene.

[0245] In some embodiments of formulas (XXI'), (XXIa'), and (XXIb'), the oxazolinated and / or oxazinylated lipids have the following structure: (hydrophobic chain)-(bifunctional moiety)-(POX and / or POZ polymer) [(hydrophobic chain)-(bifunctional moiety)] 1-2 -(alkylene moiety)-(bifunctional moiety)-(POX and / or POZ polymer) (hydrophobic chain)-(bifunctional moiety)-(cycloalkylene moiety)-(bifunctional moiety)-(POX and / or POZ polymer) (hydrophobic chain)-(bifunctional moiety)-(cycloalkenylene moiety)-(bifunctional moiety)-(POX and / or POZ polymer) (hydrophobic chain)-(bifunctional moiety)-(alkylene moiety)-(POX and / or POZ polymer) [(hydrophobic chain) 2-Contains one of -(trivalent functional moiety)]-(alkylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer).

[0246] In some embodiments of formulas (XXI’), (XXIa’), and (XXIb’), the oxazolinated and / or oxazinylated lipids are of the following formulas (XXIe’) to (XXIj’): (Hydrophobic chain)-(divalent functional moiety)-(POX and / or POZ polymer)-R 13 (XXIe’) [(Hydrophobic chain)-(divalent functional moiety)] 1-2 -(Alkylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer)-R 13 (XXIf’) (Hydrophobic chain)-(divalent functional moiety)-(cycloalkylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer)-R 13 (XXIg’) (Hydrophobic chain)-(divalent functional moiety)-(cycloalkenylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer)-R 13 (XXIh’) (Hydrophobic chain)-(divalent functional moiety)-(alkylene moiety)-(POX and / or POZ polymer)-R 13 (XXIi’) [(Hydrophobic chain 2 -(Trivalent functional moiety)]-(alkylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer)-R 13 Has one of (XXIj’).

[0247] In some embodiments of formulas (XXI’), (XXIa’), (XXIb’), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), L 11 Is [*-Z] p (C 1-6 -alkylene)-Z-, *-Z-(C 3-8 -cycloalkylene)-Z-, *-Z-(C 3-8 -cycloalkenylene)-Z-, (*=N)(C1-6 -(alkylene)-Z-, *-Z-(C 1-6 -(alkylene)-, and *-Z- selected from the group consisting of, * represents the point of attachment to R 14 , p is 1 or 2, C 1-3 -alkylene is either divalent (when p = 1) or trivalent (when p = 2), C 3-8 -cycloalkylene group and C 3-8 -cycloalkenylene group each is independently -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl and is optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of, each Z is independently -OP(O) 2 O(C 1-3 -alkylene)NH-, -NH(C 1-3 -alkylene)OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NR 22 - selected from the group consisting of, R 22 is selected from the group consisting of H, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl. For example, the linker is [*-C(O)O] p (C 1-6 -alkylene)-Z-, (*-NH)(C 1-6 -alkylene)-Z-, (*=N)(C 1-6 -alkylene)-Z-, (*-NH)C(O)(C 1-6 -alkylene)-Z-, (*-C(O)NH(C 1-6 -alkylene)-Z-, (*-NH)C(O)(C 1-6 -alkylene)-, (*-C(O)NH(C 1-6 -alkylene)-, (*-NH)C(O)-, *-C(O)NH-, *-Z-(C 3-8 -cycloalkenylene)-Z-, -S-, and -S(O) 2 - selected from the group consisting of, * represents R 14represents a junction point with, p is 1 or 2, C 1-6 -alkylene is either divalent (when p is 1) or trivalent (when p is 2), C 3-8 -cycloalkenylene group is independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and Z is -OP(O) 2 O(C 1-3 -alkylene)NH-, -NH(C 1-3 -alkylene)OP(O) 2 O-, -OC(O)NH-, -NHC(O)O-, -O-, -S-, and -NH- selected from the group consisting of.

[0248] In some embodiments of formulae (XXI’), (XXIa’), (XXIb’), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), L 11 is [*-Z] p (C 1-3 -alkylene)-Z-, *-Z-(C 3-6 -cycloalkylene)-Z-, *-Z-(C 3-6 -cycloalkenylene)-Z-, (*=N)(C 1-3 -alkylene)-Z-, *-Z(C 1-3 -alkylene)-, and *-Z- selected from the group consisting of, where * represents the junction point with R 14 and p is 1 or 2, C 1-3 -alkylene is either divalent (when p is 1) or trivalent (when p is 2), C 3-6 -cycloalkylene group and C 3-6 -cycloalkenylene group each is independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and each Z is independently -OP(O) 2 O(CH 2 ) 2NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, -N(C 1-3 -alkyl)-, and -NH- and is selected from the group consisting of. For example, the linker is [*-C(O)O] p (C 1-3 -alkylene)-Z-, (*-NH)(C 1-3 -alkylene)-Z-, (*=N)(C 1-3 -alkylene)-Z-, (*-NH)C(O)(C 1-3 -alkylene)-Z-, (*-C(O)NH(C 1-3 -alkylene)-Z-, (*-NH)C(O)(C 1-3 -alkylene)-, (*-C(O)NH(C 1-3 -alkylene)-, (*-NH)C(O)-, *-C(O)NH-, *-Z-(C 3-6 -cycloalkenylene)-Z-, -S-, and -S(O) 2 - and is selected from the group consisting of, * represents the point of attachment to R 14 , p is 1 or 2, C 1-3 -alkylene is either divalent (when p is 1) or trivalent (when p is 2), C 3-6 -cycloalkenylene group is independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and Z is -OP(O) 2 O(C 1-2 -alkylene)NH-, -NH(C 1-2 -alkylene)OP(O) 2 O-, -OC(O)NH-, -NHC(O)O-, -O-, -S-, and -NH- and is selected from the group consisting of.

[0249] In some embodiments of Formulas (XXI’), (XXIa’), (XXIb’), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), L 11 is [*-Z] p (C 1-3 -alkylene)-Z-, *-Z-(C 3-6 -cycloalkylene)-Z-, *-Z-(C 3-6 -cycloalkenylene)-Z-, (*=N)(C 1-3 -alkylene)-Z-, *-Z(C 1-3 -alkylene)-, and *-Z- selected from the group consisting of, where * represents a bond to R 14 , p is 1 or 2, C 1-3 -alkylene is either divalent (when p is 1) or trivalent (when p is 2), C 3-6 -cycloalkylene group and C 3-6 -cycloalkenylene group are each independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and each Z is independently -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, -N(C 1-3 -alkyl)-, and -NH- selected from the group consisting of. For example, the linker is [*-C(O)O] p (C 1-3 -alkylene)-Z-, (*-NH)(C 1-3 -alkylene)-Z-, (*=N)(C 1-3 -alkylene)-Z-, (*-NH)C(O)(C 1-3 -alkylene)-Z-, (*-C(O)NH(C 1-3 -alkylene)-Z-, (*-NH)C(O)(C1-3 -(alkylene)-, (*-C(O)NH(C 1-3 -(alkylene)-, (*-NH)C(O)-, *-C(O)NH-, *-Z-(C 3-6 -cycloalkenylene)-Z-, -S-, and -S(O) 2 -selected from the group consisting of, * represents the point of attachment to R 14 and p is 1 or 2, C 1-3 -alkylene is either divalent (when p is 1) or trivalent (when p is 2), C 3-6 -cycloalkenylene group is independently -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl and is optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of, and Z is -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -OC(O)NH-, -NHC(O)O-, -O-, -S-, and -NH- selected from the group consisting of.

[0250] In some embodiments of formulae (XXI’), (XXIa’), (XXIb’), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), L 11 is (*-C(O)O)(CH(OC(O)-*))(CH 2 )-Z-, (*=N)(C 1-3 -alkylene)-NHC(O)-, (*-Z)(C 1-3 -alkylene)-Z-, *-Z-(C 3-6 -cycloalkenylene)-Z-, and *-Z- selected from the group consisting of, * represents the point of attachment to R 14 and C 3-6 -cycloalkenylene group is independently -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3Optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -alkyl, and each Z is independently -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NH-; for example, the linker is (*-C(O)O)(CH(OC(O)-*))(CH 2 )-Z-, (*=N)(C 1-3 -alkylene)-NHC(O)-, (*-NH)(C 1-3 -alkylene)-NHC(O)-, *-C(O)NH-, (*-NH)C(O)-, *-Z-(C 3-6 -cycloalkenylene)-Z-, -S-, and -S(O) 2 -; * represents the point of attachment to R 14 , C 3-6 -cycloalkenylene groups are independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and Z is -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -OC(O)NH-, -NHC(O)O-, -O-, -S-, and -NH-;

[0251] In some embodiments of Formulas (XXI’), (XXIa’), (XXIb’), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), R 12 is (R 14 C(O)O)(CH(OC(O)R 14 ))(CH2 )-Z-, (R 14 ) 2 N(C 1-3 -alkylene)-NHC(O)-, R 14 Z(C 1-3 -alkylene)-Z-, R 14 Z-(C 3-6 -cycloalkenylene)-Z-, and R 14 selected from the group consisting of Z, C 3-6 -cycloalkenylene group is independently -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl and is optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of each Z is independently -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NH- and is selected from the group consisting of. For example, the linker is (R 14 C(O)O)(CH(OC(O)R 14 ))(CH 2 )-Z-, (R 14 ) 2 N(C 1-3 -alkylene)-NHC(O)-, R 14 NH(C 1-3 -alkylene)-NHC(O)-, R 14 C(O)NH-, (R 14 NH)C(O)-, R 14 Z-(C 3-6 -cycloalkenylene)-Z-, R 14 S-, and R 14 S(O) 2 - and may be selected from the group consisting of C 3-6 -cycloalkenylene group is independently -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3Optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -alkyl, and Z is -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -OC(O)NH, -NHC(O)O-, -O-, -S-, -S(O) 2 -, and -NH- selected from the group consisting of

[0252] In some embodiments of formulae (XXI’), (XXIa’), (XXIb’), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), R 12 is -L 11 (R 14 ) p i.e., the POX and / or POZ polymer is attached to one or more hydrophobic chains (i.e., R 11 ) via a linker L 14 .

[0253] In any of the above embodiments of formulae (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), (XXIb’), (XXIc), (XXId), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), each R 14 is preferably, independently, acyclic, more preferably a straight-chain hydrocarbyl group. For example, each R 14 is independently a hydrocarbyl group having at least 8 carbon atoms, e.g., at least 10 carbon atoms, preferably at most 30 carbon atoms, e.g., at most 28, 26, 24, 22, or 20 carbon atoms, or at most 16 carbon atoms, e.g., at most 15, 14, 13, 12, 11, or 10 carbon atoms. In some embodiments, each R 14 is a hydrocarbyl group having 10 to 16 carbon atoms, e.g., 10 to 15 or 10 to 14 carbon atoms. In some embodiments, each R 14is a straight-chain hydrocarbyl group having 10 to 16 carbon atoms, for example, 10 to 15 or 10 to 14 carbon atoms.

[0254] In any of the above embodiments of formulae (XXI’), (XXIa’), (XXIb’), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), each R 14 may preferably be a hydrocarbyl group having 10 to 18 carbon atoms, for example, a straight-chain alkyl group having 10 to 18 carbon atoms, or a straight-chain alkenyl group having 10 to 18 carbon atoms. For example, the straight-chain alkyl group may have 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbon atoms, and / or the straight-chain alkenyl group may have 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbon atoms and 1, 2, or 3 carbon-carbon double bonds.

[0255] In any of the above embodiments of formulae (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), (XXIb’), (XXIc), (XXId), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), R 13 is preferably H, C 1-3 alkyl, -OR 20 -N 3 C 2-6 alkynyl, -OC(O)R 21 -C(O)R 21 -NR 22 R 23 -COOH, -C(O)NR 22 R 23 -NR 22 C(O)R 21 and is selected from the group consisting of members of the targeting pair, and the C 1-3 alkyl group is independently -OH, SH, halogen, -CN, -N 3 C 2-6 alkynyl, -COOH, -NR 22 R 23 -C(O)NR 22 R23 , -NR 22 C(O)R 21 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair, R 20 is H, C 1-3 alkyl and 3- to 6-membered heterocyclyl, C 1-3 each of the alkyl and 3- to 6-membered heterocyclyl groups is independently -OH, SH, halogen, -CN, -N 3 , C 2-6 alkynyl, -COOH, -NR 22 R 23 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair, R 21 is C 1-3 alkyl and 3- to 6-membered heterocyclyl, C 1-6 each of the alkyl and 3- to 6-membered heterocyclyl groups is independently -OH, SH, halogen, -CN, -N 3 , C 2-6 alkynyl, -COOH, -NR 22 R 23 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair, R 22 and R 23 each is independently H, C 1-6 alkyl, C 2-6 alkenyl, and C 2-6 alkynyl, or R 22 and R 23 together with the nitrogen atom to which they are attached form a heterocyclyl group, C 1-6 alkyl, C 2-6 alkenyl, and C 2-6 each of the alkynyl groups is independently -OH, SH, halogen, -CN, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NH(C 1-3 alkyl), -N(C 1-3 alkyl) 2 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair.

[0256] In any of the above embodiments of Formulae (XXI), (XXI'), (XXIa), (XXIa'), (XXIb), (XXIb'), (XXIc), (XXId), (XXIe'), (XXIf'), (XXIg'), (XXIh'), (XXIi'), and (XXIj'), R 13 is preferably H, C 1-3 alkyl, -OR 20 , -N 3 , C 2-6 alkynyl, -OC(O)R 21 , -C(O)R 21 , -NR 22 R 23 , -COOH, -C(O)NR 22 R 23 , -NR 22 C(O)R 21 , and is selected from the group consisting of members of the targeting pair, and the C 1-3 alkyl group is independently -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NR 22 R 23 , -NR 22 C(O)R 21 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair, and R 20 is selected from the group consisting of H and C 1-3 alkyl, and R 21 is -OH, SH, halogen, -CN, -N 3 , C 2-6 alkynyl, -COOH, -NR 22 R 23 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair, and is C 1-3 alkyl, and each of R 22 and R 23 is independently selected from the group consisting of H, C 1-3 alkyl, C 2-3 alkenyl, and C 2-3 alkynyl, and C 1-3Alkyl, C 2-3 Alkenyl, and C 2-3 Each of the alkynyl groups is independently selected from -OH, SH, halogen, -CN, -N 3 , C 2-6 Alkynyl, -COOH, -NH 2 , -NH(C 1-3 Alkyl), -N(C 1-3 Alkyl) 2 and is optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of members of the targeting pair, or R 22 and R 23 may together with the nitrogen atom to which they are attached form a 5- or 6-membered heterocyclyl group.

[0257] In any of the above embodiments of formula (XXI), (XXI’), (XXIa), (XXIa’), (XXIb), (XXIb’), (XXIc), (XXId), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), R 13 is preferably H, C 1-3 Alkyl, -OH, -N 3 , C 2-6 Alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -C(O)NH 2 , -C(O)NHCH 3 , -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair, and the C 1-3 alkyl groups are independently -OH, -N3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 and is optionally substituted with one or more (e.g., one or two) substituents selected from the group consisting of members of the targeting pair.

[0258] In some embodiments of Formulas (XXI), (XXIa), (XXIb), (XXIc), and (XXId), R 13 is H, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), -C(O)NH 2 , -C(O)NHCH 3 , -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair. In some embodiments of Formulas (XXI), (XXIa), (XXIb), (XXIc), and (XXId), R 13 is -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 )2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), -C(O)NH 2 , -C(O)NHCH 3 , -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair. In some embodiments of Formula (XXI), (XXIa), (XXIb), (XXIc), and (XXId), R 13 is, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair. In some embodiments of Formula (XXI), (XXIa), (XXIb), (XXIc), and (XXId), R 13 is, -OH, -N 3 , -NH 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)CH3 , -NH(CH 2 CH 3 ), and -OC(O)(CH 2 ) 2 is selected from the group consisting of COOH.

[0259] In some embodiments of Formula (XXI’), (XXIa’), (XXIb’), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), R 13 is independently -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 , and optionally substituted with 1 or 2 substituents selected from the group consisting of members of the targeting pair, C 1-3 alkyl. In some embodiments of Formula (XXI’), (XXIa’), (XXIb’), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), and (XXIj’), R 13 is optionally substituted with 1 substituent selected from the group consisting of -COOH and -C(O)NH(CH 2 ) 2 NH 2 is C 1-3 alkyl.

[0260] In certain embodiments, the oxazolinated and / or oxazinylated lipid has the following general formula (XXII) or (XXII’):

Chemical formula

[0261] In some embodiments of formula (XXII) or (XXII’), a is 1, i.e., the oxazolinylated and / or oxazinylated lipid is of the following general formula (XXIIa) or (XXIIa’):

Chemical formula

[0262] In some embodiments of formula (XXII) or (XXII’), a is 2, i.e., the oxazolinylated and / or oxazinylated lipid is of the following general formula (XXIIb) or (XXIIb’):

Chemical formula

[0263] In any of the above embodiments of formula (XXIIa), (XXIIa’), (XXIIb), and (XXIIb’), R 11 , R 12 , R 13 , and m are as defined in formula (XXII) / (XXII’).

[0264] In some embodiments of Formulas (XXII), (XXII’), (XXIIa), (XXIIa’), (XXIIb), and (XXIIb’), each R 11 is methyl, or each R 11 is ethyl. In some alternative embodiments of Formulas (XXII), (XXII’), (XXIIa), (XXIIa’), (XXIIb), and (XXIIb’), R 11 is independently selected from methyl and ethyl for each repeating unit, and in at least one repeating unit, R 11 is methyl and in at least one repeating unit R 11 is ethyl.

[0265] In some embodiments of Formulas (XXII), (XXIIa), and (XXIIb), R 12 is R 14 -NHC(O)-(CH 2 )-, R 14 -NHC(O)-(CH 2 ) 2 -, -(CH 2 )-CH(OC(O)R 14 )(CH 2 OC(O)R 14 ), -(CH 2 )-CH(SR 14 )-CH 2 (SR 14 ), R 14 S-(CH 2 ) 3 -, R 14 S(O) 2 -(CH 2 ) 3 -, and R 14 -OC(O)-(CH 2 )- selected from the group consisting of, and / or R 13 is -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 )2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 )、-OC(O)(CH 2 ) 2 COOH, and selected from the group consisting of members of the target pair (e.g., R 13 is, -OH, -N 3 , -NH 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), and -OC(O)(CH 2 ) 2 COOH).

[0266] In some embodiments of formulas (XXII’), (XXIIa’), and (XXIIb’), R 12 is selected from the group consisting of R 14 C(O)NH-, R 14 S-, R 14 S(O) 2 -, and R 14 NH-(3,4-dioxocyclobut-1-ene-1,2-diyl)-NH, and / or R 13 is independently, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2) 2 NH 2 and optionally substituted with 1 or 2 substituents selected from the group consisting of a member of the targeting pair, C 1-3 alkyl (e.g., R 13 is -COOH, and -C(O)NH(CH 2 ) 2 NH 2 optionally substituted with 1 substituent selected from the group consisting of, C 1-3 alkyl).

[0267] In some embodiments, the oxazolinated and / or oxazinylated lipid has the following general formula (XXIII): R 15 -POXZ-R 16 wherein in the formula R 15 is R 17 or -L 12 (R 17 ) q and each R 17 is independently a hydrocarbyl group, L 12 is a linker, q is 1 or 2, POXZ is the following general formula (XXa) and (XXb):

Chemical formula

[0268] In some embodiments of formula (XXIII), the targeting pair is selected from the following pairs: antigen - an antibody specific for said antigen; avidin - streptavidin; folate - folate receptor; transferrin - transferrin receptor; aptamer - a molecule to which said aptamer is specific; arginine - glycine - aspartic acid (RGD) peptide - α v β 3 integrin; asparagine - glycine - arginine (NGR) peptide - aminopeptidase N; galactose - asialoglycoprotein receptor. Thus, in some embodiments of formula (XXIII), a member of the targeting pair comprises one of antigen, antibody, avidin, streptavidin, folate, transferrin, aptamer, RGD peptide, NGR peptide, and galactose.

[0269] In some of the above embodiments of formula (XXIII), R 11 can be the same alkyl group at each occurrence (i.e., in each repeating unit), for example, R 11can be methyl in each repeating unit). In some alternative embodiments of formula (XXIII), R in at least one repeating unit 11 is different from R in another repeating unit 11 (e.g., for at least one repeating unit, R 11 is one specific alkyl (e.g., ethyl), and for at least one different repeating unit, R 11 is a different specific alkyl (e.g., methyl)). For example, each R 11 can be selected from two different alkyl groups (e.g., methyl and ethyl), and not all R 1 are necessarily the same alkyl.

[0270] In some embodiments of formula (XXIII), each of R 11 is independently methyl or ethyl, preferably methyl. Thus, in some embodiments of formula (XXIII), each R 11 is methyl, or each R 11 is ethyl. In some alternative embodiments of formula (XXIII), R 11 is independently selected from methyl and ethyl for each repeating unit, and in at least one repeating unit, R 11 is methyl, and in at least one repeating unit R 11 is ethyl.

[0271] In some embodiments of formula (XXIII), the sum of the number of repeating units of formula (XXa) and the number of repeating units of formula (XXb) in the copolymer is preferably from 2 to 190, such as from 2 to 180, from 2 to 170, from 2 to 160, from 2 to 150, from 2 to 140, from 2 to 130, from 2 to 120, from 2 to 110, from 2 to 100, from 2 to 90, from 2 to 80, from 2 to 70, from 4 to 200, from 4 to 190, from 4 to 180, from 4 to 170, from 4 to 160, from 4 to 150, from 4 to 140, from 4 to 130, from 4 to 120, from 4 to 110, from 4 to 100, from 4 to 90, from 4 to 80, from 4 to 70, from 10 to 200, from 10 to 190, from 10 to 180, from 10 to 170, from 10 to 160, from 10 to 150, from 10 to 140, from 10 to 130, from 10 to 120, from 10 to 110, from 10 to 100, from 10 to 90, from 10 to 80, or from 10 to 70. In certain embodiments of formula (XXIII), the sum of the number of repeating units of formula (XXa) and the number of repeating units of formula (XXb) in the copolymer is from 2 to 180, such as from 4 to 160, from 6 to 140, from 8 to 120, or from 10 to 100, such as from 20 to 80, from 30 to 70, or from 40 to 50.

[0272] Thus, in some embodiments of formula (XXIII), the number of repeating units of formula (XXa) in the copolymer is from 1 to 179, such as from 1 to 159, from 1 to 139, from 1 to 119, or from 1 to 99, the number of repeating units of formula (XXb) in the copolymer is from 1 to 179, such as from 1 to 159, from 1 to 139, from 1 to 119, or from 1 to 99, and the sum of the number of repeating units of formula (XXa) and the number of repeating units of formula (XXb) in the copolymer is from 2 to 180, such as from 4 to 160, from 6 to 140, from 8 to 120, or from 10 to 100.

[0273] In some embodiments of formula (XXIII), L 12 is an alkylene moiety substituted with at least one functional moiety, such as at least one monovalent functional moiety, and / or the alkylene group is R 17It includes an alkylene moiety that binds to a divalent functional moiety at the end that binds to, preferably, each monovalent functional moiety is independently selected from hydroxy, ether, halogen, cyano, azide, nitro, amino, ammonium, ester, carboxyl, thiol (sulfanyl), disulfanyl, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamide, sulfonamide, sulfamate, diselenide, dithionamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbonodithioate, carbonotritioate, guanidino (imidamide), carbamimidate, carbonimidate, carbamate, carbamodithioate, carbonodithioimidate, carbamimidothioate, carbamothioate, carbonimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imide, and amide, and / or each divalent functional moiety is independently selected from ether, amino, ester, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamide, sulfonamide, sulfamate, diselenide, dithionamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imidate, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbonodithioate, carbonotritioate, guanidino (imidamide), carbamimidate, carbimidate, carbamate, carbamodithioate, carbonodithioimidate, carbamimidothioate, carbamothioate, carbimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imide, and amide.

[0274] In some embodiments of formula (XXIII), R 15 binds to the N-terminus (i.e., the terminal N atom) of the POXZ copolymer, and R 16binds to the C-terminus (i.e., the terminal C atom) of the POXZ copolymer. In some alternative embodiments of formula (XXIII), R 15 binds to the C-terminus (i.e., the terminal C atom) of the POXZ copolymer, and R 16 binds to the N-terminus (i.e., the terminal N atom) of the POXZ copolymer. The latter alternative embodiment of formula (XXIII) (i.e., R 15 binds to the C-terminus (i.e., the terminal C atom) of the POXZ copolymer, and R 16 binds to the N-terminus (i.e., the terminal N atom) of the POXZ copolymer) is shown herein as formula (XXIII’).

[0275] In some embodiments of formula (XXIII), L 12 comprises an alkylene moiety substituted with at least one monovalent functional moiety as specified above. Thus, in some embodiments, the oxazolinated and / or oxazinized lipid has the following structure (optionally, R 16 binds to the terminal C atom of the POXZ copolymer): (Hydrophobic chain) 1-2 -(An alkylene moiety substituted with at least one monovalent functional moiety)-(POXZ copolymer) and may include, wherein the “hydrophobic chain” represents R 17 , the “alkylene moiety substituted with at least one monovalent functional moiety” represents L 12 , and the “POXZ copolymer” represents the copolymer specified in formula (XXIII).

[0276] In some embodiments, the oxazolinated and / or oxazinized lipid has the following formula (XXIIIa) (optionally, R 16 binds to the terminal C atom of the POXZ copolymer): (Hydrophobic chain) 1-2 -(An alkylene moiety substituted with at least one monovalent functional moiety)-(POXZ copolymer)-R 16 and has

[0277] In some embodiments of Formulas (XXIII) and (XXIIIa), at least one monovalent functional moiety can be any one of the monovalent functional moieties specified herein, for example, hydroxy, ether, halogen, cyano, azide, nitro, amino, ammonium, ester, carboxyl, thiol (sulfanyl), disulfanyl, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamido, sulfonamido, sulfamate, diselenide, dithioamide, disulfate diamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioate, dithioate, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbonodithioate, carbonotritioate, guanidino (imidamide), carbamimidate, carbonimidate, carbamate, carbamodithioate, carbonodithioimidate, carbamimidothioate, carbamothioate, carbonimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imide, and amide.

[0278] In some embodiments of Formulas (XXIII) and (XXIIIa), the alkylene moiety substituted with at least one monovalent functional moiety is C 1-6 alkylene, for example, C 1-3 alkylene, for example, methylene, ethylene, or trimethylene.

[0279] In some embodiments of Formulas (XXIII) and (XXIIIa), the alkylene moiety substituted with at least one monovalent functional moiety is substituted with one or more (for example, from 1 to the maximum number of hydrogen atoms bonded to the alkylene moiety, for example, 1, 2, 3, 4, 5, or 6, for example, 1-5, 1-4, or 1-3, or 1 or 2) independently selected monovalent functional moieties.

[0280] In some embodiments, the alkylene moiety substituted with at least one monovalent functional moiety is C1-6-alkylene, for example, C 1-3 -alkylene, such as methylene, ethylene, or trimethylene, and is substituted with one or more (for example, from 1 to the maximum number of hydrogen atoms bonded to the alkylene moiety, such as 1, 2, 3, 4, 5, or 6, for example, 1 - 5, 1 - 4, or 1 - 3, or 1 or 2) independently selected monovalent functional moieties.

[0281] In some embodiments of formula (XXIII) (particularly those where R 16 is bonded to the terminal C atom of the POXZ copolymer), L 12 is an alkylene moiety that contains an alkylene portion that binds to the divalent functional moiety as specified above at the terminal where the alkylene group binds to R 17 . Thus, in some embodiments (particularly those where R 16 is bonded to the terminal C atom of the POXZ copolymer), the oxazolinylated and / or oxazinylated lipid has the following structure: [(hydrophobic chain)-(divalent functional moiety)] 1-2 -(alkylene portion)-(POXZ copolymer) and may include wherein "hydrophobic chain" represents R 17 and "-(divalent functional moiety)] 1-2 -(alkylene moiety)" represents L 12 and "POXZ copolymer" represents the copolymer specified in formula (XXIII).

[0282] In some embodiments (particularly those where R 16 is bonded to the terminal C atom of the POXZ copolymer), the oxazolinylated and / or oxazinylated lipid has the following formula (XXIIIb): [(hydrophobic chain)-(divalent functional moiety)] 1-2 -(alkylene portion)-(POXZ copolymer)-R 16 .

[0283] In some embodiments of formulas (XXIII) and (XXIIIb), the difunctional moiety can be any one of the difunctional moieties specified herein, for example, ether, amino, ester, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamide, sulfonamide, sulfamate, diselenide, dithioamide, disulfamide, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imidate, imino, imidothioate, thionylamide, carbonate, carbothioate, carbodithioate, carbonotrithioate, guanidino (imidamide), carbamimidate, carbodiimidate, carbamate, carbamodithioate, carbodithioimidate, carbamimidothioate, carbamothioate, carbodiimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imine, imide, and amide, and is selected from the group consisting of.

[0284] In some embodiments of formulas (XXIII) and (XXIIIb), the alkylene moiety is C 1-6 alkylene, for example, C 1-3 alkylene, for example, methylene, ethylene, or trimethylene.

[0285] In some embodiments of formula (XXIII), R 16 is bonded to the terminal N atom of the POXZ copolymer (i.e., in embodiments of formula (XXIII')), and L 12 contains at least one difunctional moiety through which one or more hydrophobic chains (R 17 ) are bonded to the POXZ copolymer. In some embodiments, L 12 is an alkylene moiety (e.g., C 1-6 alkylene moiety, e.g., C 1-3 alkylene moiety), a cycloalkylene moiety (preferably C 3-8 -cycloalkylene, e.g., C 3-6 -cycloalkylene moiety), or a cycloalkenylene moiety (preferably C3-8 -Cycloalkenylene, for example, C 3-6- It may further contain a cycloalkenylene moiety, and each of these connects a bifunctional moiety to the POXZ copolymer (either directly to the end of the POXZ copolymer or preferably via an additional bifunctional moiety). For example, one hydrophobic chain can be attached to the end of the POXZ copolymer via one bifunctional moiety (either directly, or via an alkylene, cycloalkylene, or cycloalkenylene moiety, or via an alkylene, cycloalkylene, or cycloalkenylene moiety having another bifunctional moiety), or two hydrophobic chains can be attached to the end of the POXZ copolymer via two bifunctional moieties (which preferably attaches to an alkylene, cycloalkylene, or cycloalkenylene moiety or to an alkylene, cycloalkylene, or cycloalkenylene moiety having another bifunctional moiety), or two hydrophobic chains can be attached to the end of the POXZ copolymer via the same bifunctional moiety (which is a trifunctional moiety and can be attached either directly to the end of the POXZ copolymer or via an alkylene, cycloalkylene, or cycloalkenylene moiety or via an alkylene, cycloalkylene, or cycloalkenylene moiety having another bifunctional moiety). In some embodiments, each divalent functional moiety is independently selected from ether, amino, ester, sulfide, disulfide, sulfoxide, sulfone, sulfite, sulfate, phosphate, sulfinamido, sulfonamido, sulfamate, diselenide, diamide sulfite, diamide sulfate, urea, thiourea, carbonyl, thiocarbonyl, orthoester, thioester, dithioester, imidate, imino, imidothioate, thionylamide, carbonate, carbonothioate, carbondithioate, carbonotritioate, guanidino (imidamide), carbamimidate, carbodiimidate, carbamate, carbamodithioate, carbodithioimidate, carbamimidothioate, carbamothioate, carbodiimidothioate, acylhydrazone, hydrazine, oxime, acetal, hemiacetal, ketal, hemiketal, imine, imide, and amide moieties.

[0286] In some embodiments of formula (XXIII’), the cycloalkylene moiety is C 3-8 -cycloalkylene, for example, C 3-6 -cycloalkylene, such as cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, and the cycloalkylene moiety is optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents (e.g., independently, -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl).

[0287] In some embodiments of formula (XXIII’), the cycloalkenylene moiety is C 3-8 -cycloalkenylene, for example, C 3-6 -cycloalkenylene, such as cyclopropenylene, cyclobutenylene, cyclopentenylene, cyclohexenylene, and the cycloalkenylene moiety is optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents (e.g., independently, -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl).

[0288] In some embodiments of formula (XXIII’), the alkylene moiety is C 1-6 -alkylene, for example, C 1-3 -alkylene, such as methylene, ethylene, or trimethylene, or C 2-3 alkylene.

[0289] In some embodiments of formula (XXIII’), the oxazolinized and / or oxazinized lipid has the following structure: (Hydrophobic chain)-(Difunctional moiety)-(POX and / or POZ polymer) [(Hydrophobic chain)-(Difunctional moiety)] 1-2 -(Alkylene moiety)-(Difunctional moiety)-(POX and / or POZ polymer) (Hydrophobic chain)-(divalent functional moiety)-(cycloalkylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer) (Hydrophobic chain)-(divalent functional moiety)-(cycloalkenylene moiety)-(divalent functional moiety)-(POX and / or POZ polymer) (Hydrophobic chain)-(divalent functional moiety)-(alkylene moiety)-(POX and / or POZ polymer) [(Hydrophobic chain 2 -(trivalent functional site)]-(alkylene site)-(divalent functional moiety)-(POX and / or POZ polymer) and includes one of them.

[0290] In some embodiments of formula ((XXIII’), the oxazolinated and / or oxazinized lipid is of the following formula (XXIIIc’) to (XXIIIh’): (Hydrophobic chain)-(divalent functional moiety)-(POXZ copolymer)-R 16 (XXIIIc’) [(Hydrophobic chain)-(divalent functional moiety)] 1-2 -(alkylene moiety)-(divalent functional moiety)-(POXZ copolymer)-R 16 (XXIIId’) (Hydrophobic chain)-(divalent functional moiety)-(cycloalkylene moiety)-(divalent functional moiety)-(POXZ copolymer)-R 16 (XXIIIe’) (Hydrophobic chain)-(divalent functional moiety)-(cycloalkenylene moiety)-(divalent functional moiety)-(POXZ copolymer)-R 16 (XXIIIf’) (Hydrophobic chain)-(divalent functional moiety)-(alkylene moiety)-(POXZ copolymer)-R 16 (XXIIIg’) [(Hydrophobic chain 2 -(trivalent functional part)]-(alkylene site)-(divalent functional moiety)-(POXZ copolymer)-R 16 (XXIIIh’) has one of them.

[0291] In some embodiments of formula (XXIII), (XXIIIa), and (XXIIIb), L12 comprises at least one ester, sulfide, disulfide, sulfone, orthoester, acylhydrazone, hydrazine, oxime, acetal, ketal, or amide moiety. In some embodiments, L 12 is [*-NHC(O)] q (C 1-6 -alkylene)-, [*-C(O)NH] q (C 1-6 -alkylene)-, [*-C(O)O] q (C 1-6 -alkylene)-, [*-OC(O)] q (C 1-6 -alkylene)-, [*-S] q (C 1-6 -alkylene)-, [*-SS] q (C 1-6 -alkylene)-, [*-S(O) 2 p (C 1-6 -alkylene)-, [(*-O) s C(OR 25 ) 3-s (C 1-6 -alkylene)-, [*-C(OR 25 ) 2 O] q (C 1-6 -alkylene)-, [*-C(R 25 )(=N-N(R 26 )C(O)-)] q (C 1-3 -alkylene)-, [*-C(O)(N(R 26 )-N=)C(R 25 )-] q (C 1-3 -alkylene)-, [*=C(=N-N(R 26 )C(O)(R 25 ))] q (C 1-3 -alkylene)-, [*N(R 26 )N(R 26 )] q -(C 1-6 -alkylene)-, [*=C(=N(OH))] q (C 1-6 -alkylene)-, and [*-OC(R 25 ​)(R 26 )O] q (C 1-6 -alkylene)-selected from the group consisting of, * represents the point of attachment to R 17 , q is 1 or 2, and C 1-6 -alkylene is either divalent (when q is 1) or trivalent (when q is 2), and R 25 is selected from the group consisting of C 1-6 alkyl, aryl, and aryl(C 1-6 alkyl), and R 26 is H, C 1-6 alkyl, aryl, and aryl(C 1-6 alkyl), and s is an integer from 1 to 2. For example, L 12 is [*-NHC(O)] q (C 1-3 -alkylene)-, [*-C(O)NH] q (C 1-3 -alkylene)-, [*-C(O)O] q (C 1-3 -alkylene)-, [*-OC(O)] q (C 1-3 -alkylene)-, [*-S] q (C 1-3 -alkylene)-, [*-SS] q (C 1-3 -alkylene)-, [*-S(O) 2 p(C 1-3 -alkylene)-, [(*-O) s C(OR 25 ) 3-s (C 1-3 -alkylene)-, [*-C(OR 25 ) 2 O] q (C 1-3 -alkylene)-, [*-C(R 25 )(=N-N(R 26 )C(O)-)] q (C 1-3 -alkylene)-, [*-C(O)(N(R 26 )-N=)C(R 25 )-] q (C 1-3 -alkylene)-, [*=C(=N-N(R26 )(C(O)(R 25 ))] q (C 1-3 -alkylene)-, [*N(R 26 )N(R 26 )] q (C 1-3 -alkylene)-, [*=C(=N(OH))] q (C 1-3 -alkylene)-, and [*-OC(R 25 )(R 26 )O] q (C 1-3 -alkylene)-, and may be selected from the group consisting of, * represents the point of attachment to R 17 , q is 1 or 2, C 1-3 -alkylene is either divalent (when q is 1) or trivalent (when q is 2), R 25 is selected from the group consisting of C 1-6 alkyl, aryl, and aryl(C 1-6 alkyl), R 26 is selected from the group consisting of H, C 1-6 alkyl, aryl, and aryl(C 1-6 alkyl), and s is an integer from 1 to 2.

[0292] In some embodiments of Formulas (XXIII), (XXIIIa), and (XXIIIb), R 25 is selected from the group consisting of C 1-3 alkyl, phenyl, and aryl(C 1-3 alkyl), for example, selected from the group consisting of methyl, ethyl, phenyl, benzyl, and phenylethyl.

[0293] In some embodiments of Formulas (XXIII), (XXIIIa), and (XXIIIb), R 26 is selected from the group consisting of H, C 1-3 alkyl, phenyl, and aryl(C 13 alkyl), for example, selected from the group consisting of H, methyl, ethyl, phenyl, benzyl, and phenylethyl.

[0294] In some embodiments of Formulas (XXIII), (XXIIIa), and (XXIIIb), L 12 is [*-NHC(O)] q (C 1-6 -alkylene)-, [*-C(O)NH] q (C 1-6 -alkylene)-, [*-C(O)O] q (C 1-6 -alkylene)-, [*-OC(O)] q (C 1-6 -alkylene)-, [*-S] q (C 1-6 -alkylene)-, and [*-S(O) 2 p (C 1-6 -alkylene)- selected from the group consisting of, preferably, [*-NHC(O)] q C 1-6 -alkylene)-, [*-C(O)O] q (C 1-6 -alkylene)-, [*-OC(O)] q (C 1-6 -alkylene)-, [*-S] q (C 1-6 -alkylene)-, and [*-S(O) 2 p (C 1-6 -alkylene)- selected from the group consisting of, more preferably, [*-NHC(O)] q (C 1-6 -alkylene)-, and [*-C(O)O] q (C 1-6 -alkylene)-, where * represents the point of attachment to R 17 , q is 1 or 2, and C 1-6 -alkylene is either divalent (when q = 1) or trivalent (when q = 2).

[0295] In some embodiments of Formulas (XXIII), (XXIIIa), and (XXIIIb), L 12 is [*-NHC(O)] q (C 1-3 -alkylene)-, [*-C(O)NH] q (C 1-3 ​​-(alkylene)-, [*-C(O)O] q (C 1-3 -(alkylene)-, [*-OC(O)] q (C 1-3 -(alkylene)-, [*-S] q (C 1-3 -(alkylene)-, and [*-S(O) 2 p (C 1-3 -(alkylene)- selected from the group consisting of, preferably, [*-NHC(O)] q (C 1-3 -(alkylene)-, [*-C(O)O] q C 1-3 -(alkylene)-, [*-OC(O)] q (C 1-3 -(alkylene)-, [*-S] q (C 1-3 -(alkylene)-, and [*-S(O) 2 p (C 1-3 -(alkylene)- selected from the group consisting of, more preferably, [*-NHC(O)] q (C 1-3 -(alkylene)-, and [*-C(O)O] q (C 1-3 -(alkylene)- selected from the group consisting of, where * represents the point of attachment to R 17 and q is 1 or 2, and C 1-3 -alkylene is either divalent (when q is 1) or trivalent (when q is 2).

[0296] In some embodiments of Formulas (XXIII), (XXIIIa), and (XXIIIb), L 12 is *-NHC(O)-(CH 2 )-, *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 )-, *-C(O)NH-(CH 2 ) 2 -, -(CH 2 )-CH(OC(O)-*)(CH 2 OC(O)-*), -(CH 2 )-CH(S-*) 2 ​​, -(CH 2 )-CH(S-*)-CH 2 (S-*), *-S-(CH 2 ) 3 -, *-S(O) 2 -(CH 2 ) 3 -, and *-OC(O)-(CH 2 )- selected from the group consisting of, where * represents the point of attachment to R 17 . Thus, R 15 is R 17 , -L 12 R 17 , -(CH 2 )-CH(OC(O)R 17 )(CH 2 OC(O)R 17 ), -(CH 2 )-CH(SR 17 ) 2 , and -(CH 2 )-CH(SR 17 )-CH 2 (SR 17 ) that may be selected from the group consisting of, where L 12 is *-NHC(O)-(CH 2 ), *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 ), *-C(O)NH-(CH 2 ) 2 -, *-S-(CH 2 ) 3 -, *-S(O) 2 -(CH 2 ) 3 -, and *-OC(O)-(CH 2 )- selected from the group consisting of, preferably, L 12 is *-NHC(O)-(CH 2 ), *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 ), and *-C(O)NH-(CH 2 ) 2 -, for example, *-NHC(O)-(CH 2 ), or *-NHC(O)-(CH 2 )2 selected from the group consisting of, where * represents the point of attachment to R 17 represents the point of attachment to

[0297] In some embodiments of Formulas (XXIII), (XXIIIa), and (XXIIIb), R 15 is -L 12 (R 15 ) q i.e., the POXZ copolymer is attached to one or more hydrophobic chains (i.e., R 12 ) via a linker L 17

[0298] In some embodiments of Formulas (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), L 12 is [*-Z 1 q (C 1-6 -alkylene)-Z 1 -, *-Z 1 -(C 3-8 -cycloalkylene)-Z 1 -, *-Z 1 -(C 3-8 -cycloalkenylene)-Z 1 -, (*=N)(C 1-6 -alkylene)-Z 1 -, *-Z 1 (C 1-6 -alkylene)-, and *-Z 1 -selected from the group consisting of, where * represents the point of attachment to R 17 and q is 1 or 2, C 1-6 -alkylene is either divalent (when q = 1) or trivalent (when q = 2), and each C 3-8 -cycloalkylene group and C 3-8 -cycloalkenylene group is independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and each Z 1 is independently -OP(O)​​2 O(C 1-3 -alkylene)NH-, -NH(C 1-3 -alkylene)OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NR 22 selected from the group consisting of. For example, L 12 is [*-C(O)O] q (C 1-6 -alkylene)-Z 1 (*-NH)(C 1-6 -alkylene)-Z 1 (*=N)(C 1-6 -alkylene)-Z 1 (*-NH)C(O)(C 1-6 -alkylene)-Z 1 (*-C(O)NH(C 1-6 -alkylene)-Z 1 (*-NH)C(O)(C 1-6 -alkylene)-, (*-C(O)NH(C 1-6 -alkylene)-, *-Z 1 -(C 3-8 -cycloalkenylene)-Z 1 -, *-S-, *-S(O) 2 -, (*-NH)C(O)-, and *-C(O)NH- and may be selected from the group consisting of, * represents the point of attachment to R 17 and q is 1 or 2, C 1-6 -alkylene is either divalent (when q is 1) or trivalent (when q is 2), Z 1 is -OP(O) 2 O(C 1-3 -alkylene)NH-, -NH(C 1-3 -alkylene)OP(O) 2 O-, -OC(O)NH-, -NHC(O)O-, -O-, -S-, and -NH- selected from the group consisting of.

[0299] In some embodiments of Formulas (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), L 12 is [*-Z 1 q (C 1-3 -alkylene)-Z 1 -, *-Z 1 -(C 3-6 -cycloalkylene)-Z 1 -, *-Z 1 -(C 3-6 -cycloalkenylene)-Z 1 -, (*=N)(C 1-3 -alkylene)-Z 1 -, *-Z 1 (C 1-3 -alkylene)-, and *-Z 1 -selected from the group consisting of, * represents the point of attachment to R 17 , q is 1 or 2, C 1-3 -alkylene is either divalent (when q is 1) or trivalent (when q is 2), and each C 3-6 -cycloalkylene group and C 3-6 -cycloalkenylene group is independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and each Z 1 is independently selected from the group consisting of -OP(O) 2 O(C 1-2 -alkylene)NH-, -NH(C 1-2 -alkylene)OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NR 22 . For example, L 12 is [*-C(O)O] q (C 1-3 -alkylene)-Z 1 -, (*-NH)(C 1-3 ​-alkylene)-Z 1 -, (*=N)(C 1-3 -alkylene)-Z 1 -, (*-NH)C(O)(C 1-3 -alkylene)-Z 1 -, (*-C(O)NH(C 1-3 -alkylene)-Z 1 -, (*-NH)C(O)(C 1-3 -alkylene)-, (*-C(O)NH(C 1-3 -alkylene)-, *-Z 1 -(C 3-6 -cycloalkenylene)-Z 1 -, *-S-, *-S(O) 2 -, (*-NH)C(O)-, and *-C(O)NH-, where * is R 17 represents a point of attachment to C, q is 1 or 2, 1-3 -Alkylene is either divalent (when q is 1) or trivalent (when q is 2); Z 1 -OP(O) 2 O(C 1-2 -alkylene)NH-, -NH(C 1-2 -alkylene)OP(O) 2 It is selected from the group consisting of -O-, -OC(O)NH-, -NHC(O)O-, -O-, -S-, and -NH-.

[0300] In some embodiments of formulas (XXIII'), (XXIIIc'), (XXIIId'), (XXIIIe'), (XXIIIf'), (XXIIIg'), and (XXIIIh'), L 12 is [*-Z 1 ] q (C 1-3 -alkylene)-Z 1 -, *-Z 1 -(C 3-6 -cycloalkylene)-Z 1 -, *-Z 1 -(C 3-6 -cycloalkenylene)-Z 1 -, (*=N)(C 1-3 -alkylene)-Z 1 -, *-Z 1 (C1-3 -(alkylene)-, and *-Z 1 selected from the group consisting of *, where * represents the point of attachment to R 17 and p is 1 or 2, and C 1-3 -alkylene is either divalent (when q = 1) or trivalent (when q = 2), and C 3-6 -cycloalkylene group and C 3-6 -cycloalkenylene group are each independently -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl and are optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of, and each Z 1 is independently -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, -N(C 1-3 -alkyl)-, and -NH- and is selected from the group consisting of. For example, L 12 is [*-C(O)O] q (C 1-3 -alkylene)-Z 1 -, (*-NH)(C 1-3 -alkylene)-Z 1 -, (*=N)(C 1-3 -alkylene)-Z 1 -, (*-NH)C(O)(C 1-3 -alkylene)-Z 1 -, (*-C(O)NH(C 1-3 -alkylene)-Z 1 -, (*-NH)C(O)(C 1-3 -alkylene)-, (*-C(O)NH(C 1-3 -alkylene)-, *-Z 1 -(C 3-6 -cycloalkenylene)-Z 1 -, *-S-, *-S(O) 2-、(*-NH)C(O)-、and *-C(O)NH- can be selected from the group consisting of, where * is R 17 represents the point of attachment to, p is 1 or 2, and C 1-3 -alkylene is either divalent (when q is 1) or trivalent (when q is 2), and Z 1 is -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -OC(O)NH-, -NHC(O)O-, -O-, -S-, and -NH- is selected from the group consisting of.

[0301] In some embodiments of formulae (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), L 12 is (*-C(O)O)(CH(OC(O)-*))(CH 2 )-Z 1 -, (*=N)(C 1-3 -alkylene)-NHC(O)-, (*-Z 1 (C 1-3 -alkylene)-Z 1 -, *-Z 1 -(C 3-6 -cycloalkenylene)-Z 1 -, and *-Z 1 - selected from the group consisting of, where * represents the point of attachment to R 17 , C 3-6 -cycloalkenylene group is independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and each Z 1 is independently -OP(O) 2 O(CH 2 ) 2 -NH-, -NH(CH 2 ) 2 OP(O) 2O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 is selected from the group consisting of - and -NH-. For example, L 12 is (*-C(O)O)(CH(OC(O)-*))(CH 2 )-Z 1 -, (*=N)(C 1-3 -alkylene)-NHC(O)-, (*-NH)(C 1-3 -alkylene)-NHC(O)-, *-Z 1 -(C 3-6 -cycloalkenylene)-Z 1 -, *-S-, *-S(O) 2 - and *-C(O)NH- may be selected, where * represents the point of attachment to R 17 and Z 1 is -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 -O-, -OC(O)NH-, -NHC(O)O-, -O-, -S-, and -NH-. Thus, in some embodiments of formulas (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), R 15 is (R 17 C(O)O)(CH(OC(O)R 17 ))(CH 2 )-Z 1 (R 17 ) 2 N(C 1-3 -alkylene)-Z 1 -, R 17 Z 1 (C 1-3 -alkylene)-Z 1 -, R 17 Z 1 -(C 3-6 -cycloalkenylene)-Z 1 - and R 17 Z 1 - is selected from the group consisting of - and C3-6 - The cycloalkenylene group is independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and each Z 1 is independently selected from the group consisting of -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NH-. For example, R 15 is (R 17 C(O)O)(CH(OC(O)R 17 ))(CH 2 )-Z 1 (R 17 ) 2 N(C 1-3 -alkylene)-NHC(O)-, R 17 NH(C 1-3 -alkylene)NHC(O)-, R 17 Z 1 -(C 3-6 -cycloalkenylene)-Z 1 (R 17 S-, R 17 S(O) 2 -, and R 17 C(O)NH- and may be selected from the group consisting of, Z 1 is -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NH-.

[0302] In some embodiments of formulae (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), R 15 is -L 12 (R 15 ) q i.e., the POXZ copolymer is linked to one or more hydrophobic chains (i.e., R 12 ) via a linker L 17 .

[0303] In any of the above embodiments of formulae (XXIII), (XXIIIa), (XXIIIb), (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), R 17 is preferably, independently, acyclic and preferably a straight-chain hydrocarbyl group. For example, each R 17 is preferably, independently, a hydrocarbyl group having at least 8 carbon atoms, such as at least 10 carbon atoms, preferably up to 30 carbon atoms, such as up to 28, 26, 24, 22, or 20 carbon atoms, or up to 16 carbon atoms, such as up to 15, 14, 13, 12, 11, or 10 carbon atoms. In some embodiments, each R 17 is a hydrocarbyl group having 10 to 16 carbon atoms, such as 10 to 15 or 10 to 14 carbon atoms. In some embodiments, each R 17 is a straight-chain hydrocarbyl group having 10 to 16 carbon atoms, such as 10 to 15 or 10 to 14 carbon atoms.

[0304] In any of the above embodiments of formulae (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), each R 17is preferably a hydrocarbyl group having 10 to 18 carbon atoms, for example, a straight-chain alkyl group having 10 to 18 carbon atoms, or a straight-chain alkenyl group having 10 to 18 carbon atoms. For example, the straight-chain alkenyl group may have 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbon atoms, and 1, 2, or 3 carbon-carbon double bonds.

[0305] In any of the above embodiments of Formula (XXIII), (XXIIIa), (XXIIIb), (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), R 16 is preferably H, C 1-3 alkyl, -OR 20 , -N 3 , C 2-6 alkynyl, -OC(O)R 21 , -C(O)R 21 , -NR 22 R 23 , -COOH, -C(O)NR 22 R 23 , -NR 22 C(O)R 21 , and is selected from the group consisting of members of the targeting pair, and the C 1-3 alkyl group is independently -OH, SH, halogen, -CN, -N 3 , C 2-6 alkynyl, -COOH, -NR 22 R 23 , -C(O)NR 22 R 23 , -NR 22 C(O)R 21 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair, and R 20 is selected from the group consisting of H, C 1-3 alkyl and 3- to 6-membered heterocyclyl, and each of the C 1-3 alkyl and 3- to 6-membered heterocyclyl group is independently -OH, SH, halogen, -CN, -N 3 , -COOH, -NR 22 R 23, and is optionally substituted with one or more substituents selected from the group consisting of members of the target pair, R 21 is C 1-3 selected from the group consisting of alkyl and 3- to 6-membered heterocyclyl, and each of C 1-6 alkyl and 3- to 6-membered heterocyclyl group is independently -OH, SH, halogen, -CN, -N 3 , C 2-6 alkynyl, -COOH, -NR 22 R 23 , and is optionally substituted with one or more substituents selected from the group consisting of members of the target pair, R 22 and R 23 each is independently H, C 1-6 alkyl, C 2-6 alkenyl, and C 2-6 selected from the group consisting of alkynyl, or R 22 and R 23 together with the nitrogen atom to which they are attached form a heterocyclyl group, and each of C 1-6 alkyl, C 2-6 alkenyl, and C 2-6 alkynyl group is independently -OH, SH, halogen, -CN, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NH(C 1-3 alkyl), -N(C 1-3 alkyl) 2 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair.

[0306] In any of the above embodiments of formula (XXIII), (XXIIIa), (XXIIIb), (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), R 16 is preferably H, C 1-3 alkyl, -OR 20 , -N 3 , C 2-6 alkynyl, -OC(O)R 21 , -C(O)R 21 , -NR22 R 23 ,-COOH, -C(O)NR 22 R 23 , -NR 22 C(O)R 21 , and is selected from the group consisting of members of the targeting pair, C 1-3 The alkyl group is independently -OH, -N 3 , C 2-6 Alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NR 22 R 23 , -NR 22 C(O)R 21 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair, R 20 is selected from the group consisting of H and C 1-3 Alkyl, R 21 is -OH, SH, halogen, -CN, -N 3 , C 2-6 Alkynyl, -COOH, -NR 22 R 23 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair, C 1-3 Alkyl, R 22 and R 23 each is independently selected from the group consisting of H, C 1-3 Alkyl, C 2-3 Alkenyl, and C 2-3 Alkynyl, C 1-3 Alkyl, C 2-3 Alkenyl, and C 2-3 Each of the alkynyl groups is independently -OH, SH, halogen, -CN, -N 3 , C 2-6 Alkynyl, -COOH, -NH 2 , -NH(C 13 Alkyl), -N(C 1-3 Alkyl) 2 , and is optionally substituted with one or more substituents selected from the group consisting of members of the targeting pair, or R 22 and R 23Together with the nitrogen atom to which they are attached, they can form a 5- or 6-membered heterocyclyl group.

[0307] In any of the above embodiments of formula (XXIII), (XXIIIa), (XXIIIb), (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), R 16 is preferably H, C 1-3 alkyl, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -C(O)NH 2 , -C(O)NHCH 3 , and is selected from the group consisting of members of the targeting pair, and the C 1-3 alkyl group is independently -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 , and is optionally substituted with one or more (e.g., 1 or 2) substituents selected from the group consisting of members of the targeting pair.

[0308] In some embodiments of formula (XXIII), (XXIIIa), and (XXIIIb), R16 is H, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), -C(O)NH 2 , -C(O)NHCH 3 , -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair. In some embodiments of Formulas (XXIII), (XXIIIa), and (XXIIIb), R 16 is -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), -C(O)NH 2 , -C(O)NHCH 3 , -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair. In some embodiments of Formulas (XXIII), (XXIIIa), and (XXIIIb), R 16 is -OH, -N3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair. In some embodiments of Formulas (XXIII), (XXIIIa), and (XXIIIb), R 16 is, -OH, -N 3 , -NH 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), and -OC(O)(CH 2 ) 2 COOH and is selected from the group consisting of.

[0309] In some embodiments of Formulas (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), R 16 is independently, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH2 、 -C(O)NHCH 3 、 -C(O)NH(CH 2 ) 2 NH 2 、 and is optionally substituted with 1 or 2 substituents selected from the group consisting of members of the targeting pair, C 1-3 alkyl. In some embodiments of formulas (XXIII’), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), and (XXIIIh’), R 16 is -COOH, and -C(O)NH(CH 2 ) 2 NH 2 and is optionally substituted with 1 substituent selected from the group consisting of, C 1-3 alkyl.

[0310] In certain embodiments, the oxazolinated and / or oxazinylated lipid has the following general formula (XXIV): R 15 -POXZ-R 16 wherein in the formula, R 15 , when bonded to the N-terminus of the POXZ copolymer, is -L 12 R 17 , -(CH 2 )-CH(OC(O)R 17 )(CH 2 OC(O)R 17 ), -(CH 2 )-CH(SR 17 ) 2 , and -(CH 2 )-CH(SR 17 )-CH 2 (SR 17 ), each R 17 is independently a linear hydrocarbyl group having at least 10 carbon atoms (preferably 10 to 15 carbon atoms), and L 12 is *-NHC(O)-(CH 2 ), *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2)-, *-C(O)NH-(CH 2 ) 2 -, *-S-(CH 2 ) 3 -, *-S(O) 2 -(CH 2 ) 3 -, and *-OC(O)-(CH 2 )- selected from the group consisting of (preferably, L 12 is, *-NHC(O)-(CH 2 )-, *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 )-, and *-C(O)NH-(CH 2 ) 2 -, for example, *-NHC(O)-(CH 2 )- or *-NHC(O)-(CH 2 ) 2 - selected from the group consisting of), * represents the point of attachment to R 17 , or when R 15 is attached to the C-terminus of the POXZ copolymer, (R 17 C(O)O)(CH(OC(O)R 17 ))(CH 2 )-Z 1 (R 17 ) 2 N(C 1-3 -alkylene)-Z 1 -, R 17 Z 1 (C 1-3 -alkylene)-Z 1 -, R 17 Z 1 -(C 3-6 -cycloalkenylene)-Z 1 -, and R 17 Z 1 - selected from the group consisting of, the C 3-6 cycloalkenylene group is independently optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -OH, =O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and each Z 1 is independently -OP(O) 2 O(CH2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NH- selected from the group consisting of, POXZ is a copolymer containing repeating units of the following general formulas (XXa) and (XXb):

Chemical formula

[0311] In these embodiments of formula (XXIV), R 15 is attached to the C-terminus (i.e., the terminal C atom) of the POXZ copolymer, and R 16 is attached to the N-terminus (i.e., the terminal N atom) of the POXZ copolymer), and is shown herein as formula (XXIV’).

[0312] In some embodiments of formula (XXIV) or (XXIV’), each R 11 is methyl, or each R 11 is ethyl. In some alternative embodiments of formula (XXIV) or (XXIV’), R 11 is independently selected from methyl and ethyl for each repeating unit, and in at least one repeating unit, R 11 is methyl, and in at least one repeating unit R 11 is ethyl.

[0313] In some embodiments of formula (XXIV), R 15 is attached to the N-terminus of the POXZ copolymer, and R 15 is R 17 -NHC(O)-(CH2 )-, R 17 -NHC(O)-(CH 2 ) 2 -, -(CH 2 )-CH(OC(O)R 17 )(CH 2 OC(O)R 17 ), -(CH 2 )-CH(SR 17 )-CH 2 (SR 17 ), R 17 S-(CH 2 ) 3 -, R 17 S(O) 2 -(CH 2 ) 3 -, and R 17 -OC(O)-(CH 2 )-selected from the group consisting of, and / or R 16 is -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), -OC(O)(CH 2 ) 2 COOH, and selected from the group consisting of members of the targeting pair (e.g., R 16 is -OH, -N 3 , -NH 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3)C(O)CH 3 、 -NH(CH 2 CH 3 )、 and -OC(O)(CH 2 ) 2 COOH selected from the group consisting of).

[0314] In some embodiments of formula (XXIV’) (i.e., R 15 is attached to the C-terminus of the POXZ copolymer), R 15 is, R 17 C(O)NH-, R 17 S-, R 17 S(O) 2 -, and R 17 selected from the group consisting of NH-(3,4-dioxocyclobuta-1-ene-1,2-diyl)-NH, and / or R 16 is independently, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 , and optionally substituted with one or two substituents selected from the group consisting of members of the targeting pair and is C 1-3 alkyl (e.g., R 16 is -COOH, and -C(O)NH(CH 2 ) 2 NH 2 optionally substituted with one substituent selected from the group consisting of and is C 1-3 alkyl).

[0315] In some embodiments, the oxazolinized and / or oxazinized lipid has one of the following formulas (XXV), (XXV’), (XXVI), or (XXVI’):

Chemical formula

[0316] In any of the above embodiments of Formulas (XXV), (XXV’), (XXVI), and (XXVI’), m is preferably from 10 to 100, such as from 20 to 80, from 30 to 70, or from 40 to 50, such as from 20 to 25, or from 45 to 50.

[0317] In some embodiments, the oxazolinated and / or oxazinized lipid has any one of the following Formulas (XXVa), (XXVa’), (XXVIa), or (XXVIa’):

Chemical formula

[0318] In any of the above embodiments of Formulas (XXV), (XXVa), (XXVI), and (XXVIa), R 12 is preferably -L 11 R 14 , -(CH 2 )-CH(OC(O)R 14 )(CH 2 OC(O)R 14 ), -(CH 2)-CH(SR 14 ) 2 、and -(CH 2 )-CH(SR 14 )-CH 2 (SR 14 ) selected from the group consisting of, each R 14 is independently a straight-chain hydrocarbyl group having at least 10 carbon atoms (preferably having a maximum of 30 carbon atoms, such as a maximum of 24 carbon atoms, such as 10 to 16 carbon atoms, such as 10 to 14 carbon atoms), L 11 is *-NHC(O)-(CH 2 )-, *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 )-, *-C(O)NH-(CH 2 ) 2 -, *-S-(CH 2 ) 3 -, *-S(O) 2 -(CH 2 ) 3 -, and *-OC(O)-(CH 2 )- selected from the group consisting of (preferably, L 11 is *-NHC(O)-(CH 2 )-, *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 )-, and *-C(O)NH-(CH 2 ) 2 -, for example, *-NHC(O)-(CH 2 )-, or *-NHC(O)-(CH 2 ) 2 - selected from the group consisting of), wherein * represents the point of attachment to R 14 . Accordingly, in any of the above embodiments of formula (XXV), (XXVa), (XXVI), and (XXVIa), R 12 is R 14 -NHC(O)-(CH 2 )-, R 14 -NHC(O)-(CH 2 ) 2 -, -(CH 2 )-CH(OC(O)R14 )(CH 2 O.C.(O)R 14 ), -(CH 2 )-CH(SR 14 )-CH 2 (SR 14 ), R 14 S-(CH 2 ) 3 -, R 14 S(O) 2 -(CH 2 ) 3 - and R 14 -OC(O)-(CH 2 )-.

[0319] In any of the above embodiments of formulas (XXV'), (XXVa'), (XXVI'), and (XXVIa'), R 12 is preferably (R 14 C(O)O)(CH(OC(O)R 14 ))(CH 2 )-Z-, (R 14 ) 2 N(C 1-3 -alkylene)-Z-, R 14 Z(C 1-3 -alkylene)-Z-, R 14 Z-(C 3-6 -cycloalkenylene)-Z-, and R 14 Z-, and each R 14 are independently linear hydrocarbyl groups having at least 10 carbon atoms (preferably having up to 30 carbon atoms, e.g., up to 24 carbon atoms, e.g., 10-16 carbon atoms, e.g., 10-14 carbon atoms); C 3-6 -Cycloalkenylene groups are independently -OH, ═O, -SH, halogen, -CN, -N 3 , and C 1-3 -alkyl, and each Z is independently -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O)2 Selected from the group consisting of O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NH-. Accordingly, in any of the above embodiments of formulas (XXV’), (XXVa’), (XXVI’), and (XXVIa’), R 12 is R 14 C(O)NH-, R 14 S-, R 14 S(O) 2 -, and R 14 NH-(3,4-dioxocyclobuta-1-ene-1,2-diyl)-NH- may be selected from the group consisting of

[0320] In any of the above embodiments of formulas (XXV), (XXV’), (XXVa), (XXVa’), (XXVI), (XXVI’), (XXVIa), and (XXVIa’), R 13 is preferably H, C 1-3 alkyl, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -C(O)NH 2 , -C(O)NHCH 3 , -OC(O)(CH 2 ) 2 COOH, and selected from the group consisting of members of the targeting pair, C 1-3 alkyl groups are independently -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH2 、 -NHCH 3 、 -N(CH 3 ) 2 、 -C(O)NH 2 、 -C(O)NHCH 3 、 -C(O)NH(CH 2 ) 2 NH 2 and is optionally substituted with one or more (e.g., one or two) substituents selected from the group consisting of members of the targeting pair.

[0321] In any of the above embodiments of formula (XXV), (XXVa), (XXVI), and (XXVIa), R 13 is preferably -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ) -OC(O)(CH 2 ) 2 COOH, and from the group consisting of members of the targeting pair, for example, -OH, -N 3 , -NH 2 , -NHC(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -NH(CH 2 CH 3 ), and -OC(O)(CH 2 ) 2is selected from the group consisting of -COOH.

[0322] In any of the above embodiments of formula (XXV’), (XXVa’), (XXVI’), and (XXVIa’), R 13 is preferably, independently, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 , and is C 1-3 alkyl optionally substituted with 1 or 2 substituents selected from the group consisting of members of the targeting pair, more preferably, R 13 is C 2 alkyl optionally substituted with 1 substituent selected from the group consisting of -COOH and -C(O)NH(CH 2 NH 2 . 1-3

[0323] In any of the above embodiments of formula (XXV), (XXVa), (XXVI), and (XXVIa), preferably, R 12 is -L 11 R 14 , -(CH 2 )-CH(OC(O)R 14 )(CH 2 OC(O)R 14 ), -(CH 2 )-CH(SR 14 ) 2 , and -(CH 2 )-CH(SR 14 )-CH 2 (SR 14 ), each R 14is, independently, a linear hydrocarbyl group having at least 10 carbon atoms (preferably, up to 30 carbon atoms, such as, up to 24 carbon atoms, or up to 16 carbon atoms, such as, 15, 14, 13, 12, 11, or 10 carbon atoms, such as, 10 - 16 carbon atoms, such as, 10 - 15 or 10 - 14 carbon atoms), and L 11 is, *-NHC(O)-(CH 2 )-, *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 )-, *-C(O)NH-(CH 2 ) 2 -, *-S-(CH 2 ) 3 -, *-S(O) 2 -(CH 2 ) 3 -, and *-OC(O)-(CH 2 )- selected from the group consisting of (preferably, L 11 is, *-NHC(O)-(CH 2 )-, *-NHC(O)-(CH 2 ) 2 -, *-C(O)NH-(CH 2 )-, and *-C(O)NH-(CH 2 ) 2 -, for example, *-NHC(O)-(CH 2 )-, or *-NHC(O)-(CH 2 ) 2 - selected from the group consisting of), wherein * represents the point of attachment to R 14 , R 13 is, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2 CH3 )C(O)CH 3 、-NH(CH 2 CH 3 )、-OC(O)(CH 2 ) 2 COOH, and from the group consisting of members of the targeting pair, for example, -OH, -N 3 、-NH 2 、-NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)CH 3 、-NH(CH 2 CH 3 )、and -OC(O)(CH 2 ) 2 COOH and is selected from the group consisting of.

[0324] In any of the above embodiments of formula (XXV’), (XXVa’), (XXVI’), and (XXVIa’), preferably, R 12 is (R 14 C(O)O)(CH(OC(O)R 14 ))(CH 2 )-Z-, (R 14 ) 2 N(C 1-3 -alkylene)-Z-, R 14 Z(C 1-3 -alkylene)-Z-, R 14 Z-(C 3-6 -cycloalkenylene)-Z-, and R 14 Z- and is selected from the group consisting of, each R 14 is independently a linear hydrocarbyl group having at least 10 carbon atoms (preferably having a maximum of 30 carbon atoms, for example, a maximum of 24 carbon atoms, for example, 10 to 16 carbon atoms, for example, 10 to 14 carbon atoms), and the C 3-6 -cycloalkenylene group is independently -OH, =O, -SH, halogen, -CN, -N 3 、and C 1-3Optionally substituted with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of -alkyl, and each Z is independently -OP(O) 2 O(CH 2 ) 2 NH-, -NH(CH 2 ) 2 OP(O) 2 O-, -C(O)NH-, -NHC(O)-, -OC(O)NH-, -NHC(O)O-, -O-, -C(O)O-, -OC(O)-, -S-, -S(O) 2 -, and -NH- selected from the group consisting of, R 13 is independently -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 , and optionally substituted with 1 or 2 substituents selected from the group consisting of members of the targeting pair C 1-3 alkyl, preferably, R 13 is -COOH, and -C(O)NH(CH 2 ) 2 NH 2 optionally substituted with 1 substituent selected from the group consisting of and C 1-3 alkyl.

[0325] In some embodiments, the oxazolinylated and / or oxazinylated lipid has one of the following formulas (XXVII), (XXVII'), (XXVIII), or (XXVIII'):

Chemical Formula

[0326] In any of the above embodiments of Formula (XXVII), (XXVII’), (XXVIII), or (XXVIII’), m is preferably 10 to 100, such as 20 to 80, 30 to 70, or 40 to 50, e.g., 20 to 25, or 45 to 50.

[0327] In some embodiments, the oxazolinated and / or oxazinized lipid has one of the following Formulas (XXVIIa) or (XXVIIIa):

Chemical formula

[0328] In any of the above embodiments of Formulas (XXVII), (XXVII’), (XXVIIa), (XXVIII), (XXVIII’), and (XXVIIIa), each R 14 is preferably, independently, a straight-chain hydrocarbyl group having at least 10 carbon atoms (preferably having up to 30 carbon atoms, such as up to 24 carbon atoms, such as 10 to 16 carbon atoms). In some embodiments of Formulas (XXVII), (XXVII’), and (XXVIIa), R 14 is a straight-chain hydrocarbyl group having from at least 10 to up to 16 carbon atoms, such as up to 15, 14, 13, 12, 11, or 10 carbon atoms, such as 10 to 16 carbon atoms, such as 10 to 15 or 10 to 14 carbon atoms. In some embodiments of Formulas (XXVIII), (XXVIII’), and (XXVIIIa), R 14 is, independently, a straight-chain hydrocarbyl group having from at least 10 to up to 30 carbon atoms, such as up to 24, 22, or 20 carbon atoms, such as 10 to 16 carbon atoms, such as 10 to 15 or 10 to 14 carbon atoms.

[0329] In any of the above embodiments of Formulas (XXVII), (XXVII’), (XXVIIa), (XXVIII), (XXVIII’), and (XXVIIIa), R 13 is preferably H, C 1-3 alkyl, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)CH3 、 -C(O)NH 2 、 -C(O)NHCH 3 、 -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair, and the C 1-3 alkyl group is independently -OH, -N 3 、 C 2-6 alkynyl, -COOH, -NH 2 、 -NHCH 3 、 -N(CH 3 ) 2 、 -C(O)NH 2 、 -C(O)NHCH 3 、 -C(O)NH(CH 2 ) 2 NH 2 、 and is optionally substituted with one or more (e.g., 1 or 2) substituents selected from the group consisting of members of the targeting pair.

[0330] In any of the above embodiments of formula (XXVII), (XXVII’), (XXVIIa), (XXVIII), (XXVIII’), and (XXVIIIa), preferably, each R 14 is independently a linear hydrocarbyl group having at least 10 carbon atoms (preferably, having a maximum of 30 carbon atoms, e.g., having a maximum of 24 carbon atoms, e.g., having 10 - 16 carbon atoms, e.g., having 10 - 14 carbon atoms), R 13 is H, C 1-3 alkyl, -OH, -N 3 、 C 2-6 alkynyl, -COOH, -NH 2 、 -NHCH 3 、 -N(CH 3 ) 2 、 -NH(CH 2 CH 3 )、 -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 COOH, -N(CH 2CH 3 )C(O)CH 3 、-C(O)NH 2 、-C(O)NHCH 3 、-OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair, C 1-3 The alkyl group is independently -OH, -N 3 、C 2-6 alkynyl, -COOH, -NH 2 、-NHCH 3 、-N(CH 3 ) 2 、-C(O)NH 2 、-C(O)NHCH 3 、-C(O)NH(CH 2 ) 2 NH 2 、and is optionally substituted with one or more (e.g., 1 or 2) substituents selected from the group consisting of members of the targeting pair.

[0331] Here, for formulas (XXVII’) and (XXVIII’), preferably, R 13 is -OH, -N 3 、C 2-6 alkynyl, -COOH, -NH 2 、-NHCH 3 、-N(CH 3 ) 2 、-C(O)NH 2 、-C(O)NHCH 3 、-C(O)NH(CH 2 ) 2 NH 2 、and is optionally substituted with 1 or 2 substituents selected from the group consisting of members of the targeting pair C 1-3 alkyl, and more preferably, R 13 is -COOH, and -C(O)NH(CH 2 ) 2 NH 2 alkyl optionally substituted with one substituent selected from the group consisting of 1-3 is.

[0332] In any of the above embodiments of formula (XXVII), (XXVII’), and (XXVIIa), preferably, each R 14 is independently a linear hydrocarbyl group having at least 10 carbon atoms (preferably, up to 16 carbon atoms, e.g., up to 15, 14, 13, 12, 11, or 10 carbon atoms, e.g., 10 - 16 carbon atoms, e.g., 10 - 15 or 10 - 14 carbon atoms), R 13 is H, C 1-3 alkyl, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2 , -N(CH 2 CH 3 )C(O)CH 3 , -C(O)NH 2 , -C(O)NHCH 3 , -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair, and the C 1-3 alkyl group is independently optionally substituted with one or more (e.g., 1 or 2) substituents selected from the group consisting of -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 , and members of the targeting pair.

[0333] At this position, for formula (XXVII’), preferably, R 13 is -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 , and is optionally substituted with one or two substituents selected from the group consisting of members of the targeting pair, C 1-3 alkyl, and more preferably, R 13 is -COOH, and -C(O)NH(CH 2 ) 2 NH 2 , and is optionally substituted with one substituent selected from the group consisting of, C 1-3 alkyl.

[0334] In any of the above embodiments of formula (XXVIII), (XXVIII’), and (XXVIIIa), preferably, each R 14 is independently a linear hydrocarbyl group having at least 10 carbon atoms (preferably having a maximum of 30 carbon atoms, for example, a maximum of 24 carbon atoms, for example, 10 to 15 carbon atoms), R 13 is H, C 1-3 alkyl, -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -NHC(O)(CH 2 ) 2 COOH, -N(CH 2 CH 3 )C(O)(CH 2 ) 2COOH, -N(CH 2 CH 3 )C(O)CH 3 , -C(O)NH 2 , -C(O)NHCH 3 , -OC(O)(CH 2 ) 2 COOH, and is selected from the group consisting of members of the targeting pair, C 1-3 alkyl groups are independently -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 , and is optionally substituted with one or more (e.g., 1 or 2) substituents selected from the group consisting of members of the targeting pair.

[0335] At this point, for formula (XXVIII’), preferably, R 13 is -OH, -N 3 , C 2-6 alkynyl, -COOH, -NH 2 , -NHCH 3 , -N(CH 3 ) 2 , -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NH(CH 2 ) 2 NH 2 , and is optionally substituted with 1 or 2 substituents selected from the group consisting of C 1-3 alkyl, more preferably, R 13 is C 2 alkyl optionally substituted with 1 substituent selected from the group consisting of -COOH and -C(O)NH(CH 2 NH 2 . 1-3 alkyl.

[0336] Specific examples of oxazolinated and / or oxazinylated lipids are the following compounds (XXI-1) to (XXI-34) and (XXI’-35) to (XXI’-48): [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] In each example, C 14 H 29 refers to the moiety -(CH 2 ) 13 CH 3 and, in each example, C 13 H 27 refers to the moiety -(CH 2 ) 12 CH 3 .

[0337] In some embodiments, the oxazolinated and / or oxazinized lipid is selected from any one of formulae (XXIa), (XXIa’), (XXIb), (XXIb’), (XXIc), (XXId), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), (XXIj’), (XXII), (XXII’), (XXIIa), (XXIIa’), (XXIIb), (XXIIb’), (XXIII), (XXIII’), (XXIIIa), (XXIIIb), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), (XXIIIh’), (XXIV), (XXIV’), (XXV), (XXV’), (XXVa), (XXVa’), (XXVI), (XXVI’), (XXVIa), (XXVIa’), (XXVII), (XXVII’), (XXVIIa), (XXVIII), (XXVIII’), (XXVIIIa), (XXI-1), (XXI-2), (XXI-3), (XXI-4), (XXI-5), (XXI-6), (XXI-7), (XXI-8), (XXI-9), (XXI-10), (XXI-11), (XXI-12), (XXI-13), (XXI-14), (XXI-15), (XXI-16), (XXI-17), (XXI-18), (XXI-19), (XXI-20), (XXI-21), (XXI-22), (XXI-23), (XXI-24), (XXI-25), (XXI-26), (XXI-27), (XXI-28), (XXI-29), (XXI-30), (XXI-31), (XXI-32), (XXI-33), (XXI-34), (XXI’-35), (XXI’-36), (XXI’-37), (XXI’-38), (XXI’-39), (XXI’-40), (XXI’-41), (XXI’-42), (XXI’-43), (XXI’-44), (XXI’-45), (XXI’-46), (XXI’-47), and (XXI’-48).

[0338] In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) may include the cationic / cationizable lipids described herein (e.g., the cationic cationizable lipids of formula (I), (X), or (XI)), the oxazolinylated and / or oxazinylated lipids described herein, the phospholipids described herein, and cholesterol, and the oxazolinylated and / or oxazinylated lipids described herein are preferably of formula (XXIa), (XXIa’), (XXIb), (XXIb’), (XXIc), (XXId), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), (XXIj’), (XXII), (XXII’), (XXIIa), (XXIIa’), (XXIIb), (XXIIb’), (XXIII), (XXIII’), (XXIIIa), (XXIIIb), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), (XXIIIh’), (XXIV), (XXIV’), (XXV), (XXV’), (XXVa), (XXVa’), (XXVI), (XXVI’), (XXVIa), (XXVIa’), (XXVII), (XXVII’), (XXVIIa), (XXVIII), (XXVIII’), (XXVIIIa), (XXI-1), (XXI-2), (XXI-3), (XXI-4), (XXI-5), (XXI-6), (XXI-7), (XXI-8), (XXI-9), (XXI-10), (XXI-11), (XXI-12), (XXI-13), (XXI-14), (XXI-15), (XXI-16), (XXI-17), (XXI-18), (XXI-19), (XXI-20), (XXI-21), (XXI-22), (XXI-23), (XXI-24), (XXI-25), (XXI-26), (XXI-27), (XXI-28), (XXI-29), (XXI-30), (XXI-31), (XXI-32), (XXI-33), (XXI-34), (XXI’-35), (XXI’-36), (XXI’-37), (XXI’-38), (XXI’-39), (XXI’-40), (XXI’-41), (XXI’-42), (XXI’-43), (XXI’-44), (XXI’-45),Is any one of, or includes, (XXI’-46), (XXI’-47), and (XXI’-48).

[0339] In some embodiments, the oxazolinylated and / or oxazinylated lipids are present at about 0.1 mol% to about 10 mol%, such as about 0.2 mol% to about 9 mol%, about 0.5 mol% to about 8 mol%, about 1 mol% to about 7.5 mol%, about 1.5 mol% to about 7 mol%, about 2 mol% to about 6.5 mol%, about 2.5 mol% to about 6 mol%, or about 3 mol% to about 5 mol% of the total lipids present in the nucleic acid compositions (e.g., DNA or RNA compositions, particularly mRNA compositions) described herein.

[0340] In some embodiments, when at least a portion of (i) a nucleic acid (e.g., DNA or RNA, particularly mRNA), (ii) a cationic / cationizable lipid, and (iii) an oxazolinylated and / or oxazinylated lipid forms particles (e.g., nanoparticles such as LNPs), the oxazolinylated and / or oxazinylated lipids can be present at about 0.1 mol% to about 10 mol%, such as about 0.2 mol% to about 9 mol%, about 0.5 mol% to about 8 mol%, about 1 mol% to about 7.5 mol%, about 1.5 mol% to about 7 mol%, about 2 mol% to about 6.5 mol%, about 2.5 mol% to about 6 mol%, or about 3 mol% to about 5 mol% of the total lipids present in the particles.

[0341] In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) can include the cationic / cationizable lipids described herein (e.g., the cationic cationizable lipids of formula (X) or (XI)), the oxazolinylated and / or oxazinylated lipids described herein, phospholipids, and cholesterol, wherein the cationic / cationizable lipid is included at about 40 mol% to about 50 mol% of the total lipids present in the composition, the oxazolinylated and / or oxazinylated lipid is included at about 0.5 mol% to about 10 mol% (e.g., about 2 mol to about 5 mol%) of the total lipids present in the composition, the phospholipid is included at about 5 mol% to about 15 mol% of the total lipids present in the composition, and cholesterol is included at about 30 mol% to about 50 mol% of the total lipids present in the composition.In some embodiments, the oxazolinated and / or oxazinized lipid has any one of formulas (XXIa), (XXIa’), (XXIb), (XXIb’), (XXIc), (XXId), (XXIe’), (XXIf’), (XXIg’), (XXIh’), (XXIi’), (XXIj’), (XXII), (XXII’), (XXIIa), (XXIIa’), (XXIIb), (XXIIb’), (XXIII), (XXIII’), (XXIIIa), (XXIIIb), (XXIIIc’), (XXIIId’), (XXIIIe’), (XXIIIf’), (XXIIIg’), (XXIIIh’), (XXIV), (XXIV’), (XXV), (XXV’), (XXVa), (XXVa’), (XXVI), (XXVI’), (XXVIa), (XXVIa’), (XXVII), (XXVII’), (XXVIIa), (XXVIII), (XXVIII’), (XXVIIIa), (XXI-1), (XXI-2), (XXI-3), (XXI-4), (XXI-5), (XXI-6), (XXI-7), (XXI-8), (XXI-9), (XXI-10), (XXI-11), (XXI-12), (XXI-13), (XXI-14), (XXI-15), (XXI-16), (XXI-17), (XXI-18), (XXI-19), (XXI-20), (XXI-21), (XXI-22), (XXI-23), (XXI-24), (XXI-25), (XXI-26), (XXI-27), (XXI-28), (XXI-29), (XXI-30), (XXI-31), (XXI-32), (XXI-33), (XXI-34), (XXI’-35), (XXI’-36), (XXI’-37), (XXI’-38), (XXI’-39), (XXI’-40), (XXI’-41), (XXI’-42), (XXI’-43), (XXI’-44), (XXI’-45), (XXI’-46), (XXI’-47), and (XXI’-48).

[0342] In some embodiments, the polymer-conjugated lipid is about 0.5 to about 5 mol% based on the total lipids in the LNP. In some embodiments, the LNP comprises about 1.0 to about 2.5 mol% of the polymer-conjugated lipid. In some embodiments, the LNP comprises about 1.5 to about 2.0 mol% of the polymer-conjugated lipid. In some embodiments, the LNP comprises about 1.5 to about 1.8 mol% of the polymer-conjugated lipid.

[0343] In some embodiments, the molar ratio of the total cationic lipid to the total polymer-conjugated lipid (e.g., PEG-conjugated lipid) is from about 100:1 to about 20:1. In some embodiments, the molar ratio of the total cationic lipid to the total polymer-conjugated lipid (e.g., PEG-conjugated lipid) is from about 50:1 to about 20:1. In some embodiments, the molar ratio of the total cationic lipid to the total polymer-conjugated lipid (e.g., PEG-conjugated lipid) is from about 40:1 to about 20:1. In some embodiments, the molar ratio of the total cationic lipid to the total polymer-conjugated lipid (e.g., PEG-conjugated lipid) is from about 35:1 to about 25:1.

[0344] Immunomodulator As described herein, the lipid nanoparticles comprise an immunomodulator. In some embodiments, the immunomodulator is an immunosuppressant. In some embodiments, the immunomodulator is an immunostimulant. In some embodiments, the immunomodulator is an agent (e.g., a small molecule) that is an agonist or antagonist of a toll-like receptor (e.g., TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10) or a pattern recognition receptor (PRR). In some embodiments, the immunomodulator is an sp 2 -iminosugar glycolipid. In some embodiments, the immunomodulator is a small molecule downstream inhibitor of NF-κβ. In some embodiments, the immunomodulator is an sp 2 -iminosugar glycolipid. In some embodiments, the immunomodulator is an inflammasome inhibitor.

[0345] In some embodiments, the lipid nanoparticles described herein do not contain dexamethasone or a dexamethasone prodrug (e.g., dexamethasone conjugated with a biodegradable linker).

[0346] In some embodiments, the immunomodulatory agent is a TLR inhibitor. In some embodiments, the TLR inhibitor is an inhibitor of TLR2, TLR4, and / or TLR6. In some embodiments, the TLR inhibitor is an inhibitor of TLR2. In some embodiments, the immunomodulatory agent is an inhibitor of TLR4. In some embodiments, the immunomodulatory agent is an inhibitor of TLR6. In some embodiments, the inhibitor of TLR4 is TAK-242:

Chemical formula

[0347] In some embodiments, the immunomodulatory agent is a terpenoid. In some embodiments, the terpenoid is a triterpene. In some embodiments, the immunomodulatory agent is a triterpenoid. In some embodiments, the triterpenoid is a synthetic or natural derivative of amyrin, betulinic acid, oleanolic acid, sterol, squalene, or ursolic acid.

[0348] In some embodiments, the immunomodulatory agent is a corticosteroid. In some embodiments, the corticosteroid is a glucocorticoid. In some embodiments, the glucocorticoid is selected from dexamethasone, prednisone, fluticasone propionate, budesonide, or a pharmaceutically acceptable salt thereof. In some embodiments, the glucocorticoid is dexamethasone, or a pharmaceutically acceptable salt thereof. In some embodiments, the glucocorticoid is prednisone, or a pharmaceutically acceptable salt thereof. In some embodiments, the glucocorticoid is fluticasone, or a pharmaceutically acceptable salt thereof. In some embodiments, the glucocorticoid is propionate, or a pharmaceutically acceptable salt thereof. In some embodiments, the glucocorticoid is budesonide, or a pharmaceutically acceptable salt thereof.

[0349] In some embodiments, the immunomodulatory agent is an inflammasome inhibitor. In some embodiments, the inflammasome inhibitor is an NLRP3 inflammasome inhibitor, an AIM2 inflammasome inhibitor, a caspase-1 inhibitor, or a pan-caspase inhibitor. In some embodiments, the inflammasome inhibitor is selected from glibride (i.e., glibenclamide), 5-chloro-2-methoxy-N-[2-(4-sulfamoylphenyl)-ethyl]-benzamide (i.e., 16673-34-0), JC124, FC11A-2, parthenolide, VX-740, VX-765, BAY 11-7082, BHB, MCC950, MNS, CY-09, tranilast, OLT1177, and oridonin. In some embodiments, the inflammasome inhibitor is glibride (i.e., glibenclamide).

Chemical formula

[0350] In some embodiments, the inflammasome inhibitor is 5-chloro-2-methoxy-N-[2-(4-sulfamoylphenyl)-ethyl]-benzamide (i.e., 16673-34-0).

Chemical formula

[0351] In some embodiments, the inflammasome inhibitor is JC124.

Chemical formula

[0352] In some embodiments, the inflammasome inhibitor is FC11A-2.

Chemical formula

[0353] In some embodiments, the inflammasome inhibitor is parthenolide.

Chemical formula

[0354] In some embodiments, the inflammasome inhibitor is VX-740.

Chemical formula

[0355] In some embodiments, the inflammasome inhibitor is VX-765.

Chemical formula

[0356] In some embodiments, the inflammasome inhibitor is BAY 11-7082.

Chemical formula

[0357] In some embodiments, the inflammasome inhibitor is beta-hydroxybutyrate (BHB).

[0358] In some embodiments, the inflammasome inhibitor is MCC950.

Chemical formula

[0359] In some embodiments, the inflammasome inhibitor is 3,4-methylenedioxy-β-nitrostyrene (MNS).

[0360] In some embodiments, the inflammasome inhibitor is CY-09.

Chemical formula

[0361] In some embodiments, the inflammasome inhibitor is tranilast.

Chemical formula

[0362] In some embodiments, the inflammasome inhibitor is OLT1177.

Chemical formula

[0363] In some embodiments, the inflammasome inhibitor is oridonin.

Chemical formula

[0364] In some embodiments, the lipid nanoparticles described herein contain more than one immunomodulatory agent. In some embodiments, the lipid nanoparticles contain an immunomodulatory agent and one or more additional immunomodulatory agents. In some embodiments, the lipid nanoparticles contain a terpenoid (e.g., a corticosteroid, a glucocorticoid, e.g., dexamethasone, prednisone, fluticasone propionate, or budesonide, or an inflammasome inhibitor), and one or more additional immunomodulatory agents (e.g., a corticosteroid, or a glucocorticoid, e.g., another one of dexamethasone, prednisone, fluticasone propionate, or budesonide, or a TLR or PRR agonist or antagonist described herein, or an inflammasome inhibitor). In some embodiments, the lipid nanoparticles contain an immunomodulatory agent that is a terpenoid (including any subclass described herein), and a small molecule agonist or antagonist of TLR or PRR (including any subclass described herein, such as TAK-242).

[0365] In some embodiments, the lipid nanoparticles described herein contain a cationic lipid and an immunomodulatory agent, and the immunomodulatory agent is a TLR inhibitor. In some embodiments, the lipid nanoparticles contain a cationic lipid and an immunomodulatory agent, and the immunogenic agent is an inflammasome inhibitor. In some embodiments, the lipid nanoparticles contain a cationic lipid, RNA, and an immunomodulatory agent selected from TAK-242, MCC950, and BAY 11-7082. In some embodiments, the lipid nanoparticles contain a cationic lipid, RNA, and TAK-242. In some embodiments, the lipid nanoparticles contain a cationic lipid, RNA, and MCC950. In some embodiments, the lipid nanoparticles contain a cationic lipid, RNA, and BAY 11-7082.

[0366] In some embodiments, the lipid nanoparticles comprise from about 0.1 to about 50 mol% immunomodulatory agent (relative to the total amount of lipid and immunomodulatory agent). In some embodiments, the lipid nanoparticles comprise from about 1 to about 50 mol% immunomodulatory agent (relative to the total amount of lipid and immunomodulatory agent). In some embodiments, the lipid nanoparticles comprise from about 30 to about 50 mol% immunomodulatory agent (relative to the total amount of lipid and immunomodulatory agent). In some embodiments, the lipid nanoparticles comprise from about 30 to about 50 mol% immunomodulatory agent (relative to the total amount of lipid and immunomodulatory agent). In some embodiments, the lipid nanoparticles comprise from about 30 to about 35 mol% immunomodulatory agent (relative to the total amount of lipid and immunomodulatory agent). In some embodiments, the lipid nanoparticles comprise from about 35 to about 45 mol% immunomodulatory agent (relative to the total amount of lipid and immunomodulatory agent). In some embodiments, the lipid nanoparticles comprise from about 40 to about 50 mol% immunomodulatory agent (relative to the total amount of lipid and immunomodulatory agent). In some embodiments, the lipid nanoparticles comprise about 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 mol% immunomodulatory agent (relative to the total amount of lipid and immunomodulatory agent).

[0367] In some embodiments, the nucleic acid particles comprise: i) from about 30 to about 50 mol% cationic lipid; ii) from about 1 to about 5 mol% polymer-conjugated lipid; iii) from about 5 to about 15 mol% helper lipid; and iv) an immunomodulatory agent. In some embodiments, the nucleic acid particles comprise: i) from about 30 to about 50 mol% ALC-0315; ii) from about 1 to about 5 mol% ALC-0159; iii) from about 5 to about 15 mol% DSPC; and iv) an immunomodulatory agent. In some embodiments, the nucleic acid particles comprise: i) from about 30 to about 50 mol% ALC366; ii) from about 1 to about 5 mol% ALC-0159; iii) from about 5 to about 15 mol% DSPC; and iv) an immunomodulatory agent.

[0368] In some embodiments, the nucleic acid particle comprises: i) about 47.5 mol% of ALC-0315; ii) about 1.8 mol% of ALC-0159; and iii) about 10 mol% of DSPC. In some embodiments, the nucleic acid particle comprises: i) about 47.5 mol% of ALC366; ii) about 1.8 mol% of ALC-0159; and iii) about 10 mol% of DSPC.

[0369] In some embodiments, the nucleic acid particle comprises: i) about 30 to about 50 mol% of SM-102; ii) about 1 to about 5 mol% of PEG2000-DMG; iii) about 5 to about 15 mol% of DSPC; and iv) an immunomodulatory agent. In some embodiments, the nucleic acid particle comprises: i) about 50 mol% of SM-102; ii) about 1.5 mol% of PEG2000-DMG; iii) about 10 mol% of DSPC; and iv) an immunomodulatory agent. In some embodiments, the immunomodulatory agent is TAK-242, MCC950, or BAY11-7082. In some embodiments, the nucleic acid particle comprises: i) about 50 mol% of SM-102; ii) about 1.5 mol% of PEG2000-DMG; iii) about 10 mol% of DSPC; and iv) TAK-242. In some embodiments, the nucleic acid particle comprises: i) about 50 mol% of SM-102; ii) about 1.5 mol% of PEG2000-DMG; iii) about 10 mol% of DSPC; and iv) MCC950. In some embodiments, the nucleic acid particle comprises: i) about 50 mol% of SM-102; ii) about 1.5 mol% of PEG2000-DMG; iii) about 10 mol% of DSPC; and iv) BAY 11-7082.

[0370] Terpenoid As described herein, in some embodiments, the nucleic acid particles further comprise a terpenoid. In some embodiments, the terpenoid is a triterpene. In some embodiments, the terpenoid is a steroid. In some embodiments, the steroid is a sterol. In some embodiments, the sterol is β-sitosterol, stigmasterol, cholesterol, cholecalciferol, ergocalciferol, calcipotriol, botulin, lupeol, ursolic acid, oleanolic acid, cycloartenol, lanosterol, or α-tocopherol. In some embodiments, the sterol is β-sitosterol. In some embodiments, the sterol is stigmasterol. In some embodiments, the sterol is cholesterol. In some embodiments, the sterol is cholecalciferol. In some embodiments, the sterol is ergocalciferol. In some embodiments, the sterol is calcipotriol. In some embodiments, the sterol is botulin. In some embodiments, the sterol is lupeol. In some embodiments, the sterol is ursolic acid. In some embodiments, the sterol is oleanolic acid. In some embodiments, the sterol is cycloartenol. In some embodiments, the sterol is lanosterol. In some embodiments, the sterol is α-tocopherol.

[0371] In some embodiments, the lipid nanoparticles comprise from about 30 to about 50 mol% of a steroid. In some embodiments, the lipid nanoparticles comprise from about 35 to about 45 mol% of a steroid. In some embodiments, the lipid nanoparticles comprise from about 38 to about 40 mol% of a steroid. In some embodiments, the lipid nanoparticles comprise about 38.5 mol% of a steroid. In some embodiments, the lipid nanoparticles comprise about 40 mol% of a steroid.

[0372] In some embodiments, the lipid nanoparticles contain from about 30 to about 50 mol% cholesterol. In some embodiments, the lipid nanoparticles contain from about 35 to about 45 mol% cholesterol. In some embodiments, the lipid nanoparticles contain from about 38 to about 41 mol% cholesterol. In some embodiments, the lipid nanoparticles contain about 38.5 mol% cholesterol. In some embodiments, the lipid nanoparticles contain about 40.7 mol% cholesterol.

[0373] Method for producing lipid nanoparticles Lipids and lipid nanoparticles containing nucleic acids, and methods for their preparation are described, for example, in U.S. Patent Nos. 8,569,256 and 5,965,No. 542, as well as U.S. Patent Publication Nos. 2016 / 0199485, 2016 / 0009637, 2015 / 0273068, 2015 / 0265708, 2015 / 0203446, 2015 / 0005363, 2014 / 0308304, 2014 / 0200257, 2013 / 086373, 2013 / 0338210, 2013 / 0323269, 2013 / 0245107, 2013 / 0195920, 2013 / 0123338, 2013 / 0022649, 2013 / 0017223, 2012 / 0295832, 2012 / 0183581, 2012 / 0172411, 2012 / 0027803, 2012 / 0058188, 2011 / 0311583, 2011 / 0311582, 2011 / 0262527, 2011 / 0216622, 2011 / 0117125, 2011 / 0091525, 2011 / 0076335, 2011 / 0060032, 2010 / 0130588, 2007 / 0042031, 2006 / 0240093, 2006 / 0083780, 2006 / 0008910, 2005 / 0175682, 2005 / 017054, 2005 / 0118253, 2005 / 0064595, 2004 / 0142025, 2007 / 0042031, 1999 / 009076, as well as PCT Publication Nos. WO99 / 39741, WO2018 / 081480, WO2017 / 004143, WO2017 / 075531, WO2015 / 199952, WO2014 / 008334, WO2013 / 086373, WO2013 / 086322, WO2013 / 016058, WO2013 / 086373, W02011 / 141705, WO2001 / 07548, etc., which are known in the art, and the entire disclosure of which is hereby incorporated by reference in its entirety for the purposes described herein.,

[0374] For example, in some embodiments, the cationic lipid, helper lipid, and immunomodulator are solubilized in ethanol at a predetermined weight or molar ratio / percentage (e.g., as described herein). In some embodiments, the lipid nanoparticles (LNPs) are prepared at an approximate total lipid to RNA weight ratio of 10:1 to 30:1. In some embodiments, such RNA can be diluted to 0.2 mg / mL in acetate buffer.

[0375] In some embodiments, using the ethanol injection technique, a colloidal lipid dispersion containing RNA can be formed as follows: an ethanol solution containing lipids, such as a cationic lipid, helper lipid, immunomodulator, and polymer-conjugated lipid, is injected into an aqueous solution containing RNA (e.g., as described herein).

[0376] In some embodiments, the lipid and RNA solutions can be mixed at room temperature by pumping each solution (e.g., the lipid solution containing the cationic lipid, helper lipid, immunomodulator, and any other additives) into a mixing unit at a controlled flow rate, for example, using a piston pump. In some embodiments, the flow rates of the lipid solution and the RNA solution into the mixing unit are maintained at a ratio of 1:3. Upon mixing, the ethanol lipid solution is diluted with aqueous RNA, and nucleic acid-lipid particles are formed. The lipid solubility decreases, and the cationic lipid having a positive charge interacts with the negatively charged RNA.

[0377] In some embodiments, the solution containing RNA-encapsulated lipid nanoparticles can be processed by one or more of concentration adjustment, buffer exchange, formulation, and / or filtration.

[0378] Polymeric particles In some embodiments, the nucleic acid particles described herein comprise a cationic polymer, RNA, and an immunomodulatory agent. Examples of nucleic acid particles suitable for RNA delivery are described in PCT Application Publication No. WO2021 / 001417, which is hereby incorporated by reference in its entirety. Generally as described above, the complexes described herein comprise a cationic polymer. In some embodiments, the cationic polymer is a polycationic polymer, e.g., a polymer having one or more cationic or ionizable groups. As described herein, a "cationic" group is a group having a net positive charge. As used herein, an "ionizable" group is a group that can have a neutral charge at a particular pH, but can become charged (e.g., cationic) at a different pH. For example, in some embodiments, the ionizable group becomes cationic (i.e., positively charged) at physiological pH (e.g., pH of about 7.4). Thus, reference to a cationic polymer is understood to refer to both the neutral form and the charged (i.e., ionic) form. One of ordinary skill in the art will also understand that the cationic groups described herein can also exist as salts (e.g., pharmaceutically acceptable salts) comprising a cationic group and one or more suitable counterions. For example, in some embodiments, the cationic groups described herein comprise ammonium chloride. Suitable counterions include halogens (e.g., Br - , Cl - , I - , F - ), acetate (e.g., C(O)O - ), and the like. For additional examples, see the definition of pharmaceutically acceptable salts described herein.

[0379] In some embodiments, one or more cationic or ionizable groups comprise a nitrogen atom. Cationic polymers useful for preparing the complexes described herein can be homopolymers, heteropolymers, or block copolymers.

[0380] In some embodiments, the cationic polymer is poly(ethyleneimine), ...

Claims

1. A nucleic acid particle comprising RNA, an immunomodulator, and a cationic lipid or cationic polymer, wherein the immunomodulator is selected from the group consisting of inflammasome inhibitors, TLR inhibitors, small molecule downstream inhibitors of NF-κβ, sp2-iminoglycolipids, and glucocorticoids.

2. The nucleic acid particle according to claim 1, wherein the immunomodulator is 0.001 to 50 mol%, or 0.01 to 0.09 mol%, of the total amount of lipids or polymers and immunomodulators in the nucleic acid particle.

3. (a) The TLR inhibitor is an inhibitor of TLR2, TLR4, and / or TLR6, optionally the TLR inhibitor is an inhibitor of TLR4, optionally the TLR inhibitor is TAK-242, (b) The glucocorticoid is prednisolone, fluticasone propionate, budesonide, or a pharmaceutically acceptable salt thereof, or (c) The nucleic acid particle according to claim 1, wherein the inflammasome inhibitor is an NLRP3 inflammasome inhibitor, an AIM2 inflammasome inhibitor, a caspase-1 inhibitor, or a pancassase inhibitor, and optionally the inflammasome inhibitor is selected from glybride (e.g., glibenclamide), 5-chloro-2-methoxy-N-[2-(4-sulfamoylphenyl)-ethyl]-benzamide (e.g., 16673-34-0), JC124, FC11A-2, parthenolide, VX-740, VX-765, BAY 11-7082, BHB, MCC950, MNS, CY-09, tranilast, OLT1177, and oridonin, and further optionally the inflammasome inhibitor is MCC950 or BAY 11-7082.

4. The lipid nanoparticles further comprise one or more additional immunomodulators, optionally, the one or more additional immunomodulators (i) Small molecule agonists or antagonists of TLR or PRR receptors; or (ii) Inflammasome inhibitors The nucleic acid particles according to claim 1, which are or include them.

5. The nucleic acid particle according to claim 1, wherein the nucleic acid particle comprises the immunomodulator, a cationic lipid, and RNA.

6. The nucleic acid particle according to claim 5, wherein the nucleic acid particle is in the form of a lipid nanoparticle.

7. (a) The cationic lipid is a lipid containing one or more nitrogen atoms that are cationic or ionizable at a physiological pH (e.g., 7.4), or (b) The cationic lipid is one of the following compounds: Table 1 Table 2 Table 3 Table 4 Table 5 Table 6 Table 7 Table 8 Table 9 Table 10 Table 11 Table 12 Table 13 Table 14 Table 15 Or selected from those pharmaceutically acceptable salts, (c) The cationic lipid is one of the following compounds: Table 16 Table 17 Table 18 Table 19 Table 20 Or selected from those pharmaceutically acceptable salts, Optionally, the cationic lipid is 【Chemistry 1】 N,N-dimethyl-2,3-dioleoyloxypropylamine (DODMA), or 【Chemistry 2】 The nucleic acid particle according to claim 1.

8. (a) The cationic lipid is 30 to 50 mol% of the total amount of lipids and immunomodulators in the lipid nanoparticles, and / or (b) The nucleic acid particles further comprise a helper lipid, optionally the helper lipid being a phospholipid, and the helper lipid being selected from phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, and sphingomyelin, more preferably distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), dipentadecanoylphosphatidylcholine, dilauroylphosphatidylcholine, dipalmitoylphospha Tidylcholine (DPPC), Diarachidoylphosphatidylcholine (DAPC), Dibehenoylphosphatidylcholine (DBPC), Ditricosanoylphosphatidylcholine (DTPC), Dilignoceroylphatidylcholine (DLPC), Palmitoyloleoylphosphatidylcholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 dietherPC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 A helper lipid is selected from the group consisting of Lyso PC, dioleoyl phosphatidylethanolamine (DOPE), distearoyl phosphatidylethanolamine (DSPE), dipalmitoyl phosphatidylethanolamine (DPPE), dimyristoyl phosphatidylethanolamine (DMPE), dilauroyl phosphatidylethanolamine (DLPE), diphytanoyl phosphatidylethanolamine (DPyPE), and combinations thereof, preferably the helper lipid is DSPC. Optionally, the lipid nanoparticles contain 5 to 15 mol% of the helper lipids relative to the total amount of lipids and immunomodulators, and / or (c) The nucleic acid particles further comprise polymer-conjugated lipids, optionally the polymer-conjugated lipids being PEG lipids selected from PEG-DAG, PEG-PE, PEG-S-DAG, PEG2000-DMG, PEG-Cer, PEG-dialkyloxypropyl carbamate, ALC-0159, and combinations thereof, preferably the PEG lipids being ALC-0159 or PEG2000-DMG. Optionally, the lipid nanoparticles contain 1 to 5 mol% of the PEG lipid relative to the total amount of lipids and immunomodulators, and / or (d) The nucleic acid particles further comprise a terpenoid, optionally the terpenoid being a triterpene, the triterpene being a sterol, and the sterol being selected from β-sitosterol, stigmasterol, cholesterol, cholecalciferol, ergocalciferol, calcipotriol, botulinum, lupeol, ursolic acid, oleanolic acid, cycloartenol, lanosterol, or α-tocopherol, and / or (e) The nucleic acid particles are compounds comprising an amphiphilic moiety selected from polysorbate, poloxamer, and / or polyalkylene glycol (e.g., polyethylene glycol), poly(2-oxazoline), poly(2-oxazine), polysarcosine, polyvinylpyrrolidone, and poly[N-(2-hydroxypropyl)methacrylamide], or comprising the same, further comprising a surfactant wherein the amphiphilic moiety is bonded to one or more C12-C20 aliphatic groups, The nucleic acid particles according to claim 6, wherein optionally, the surfactant is a polysorbate selected from polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, and combinations thereof, or comprises them.

9. (a) The cationic lipid is ALC-0315, and the lipid nanoparticles further include a helper lipid that is DSPC and a PEG lipid that is ALC-0159, or (b) The cationic lipid is SM-102, and the lipid nanoparticles further comprise a helper lipid which is DSPC and a PEG lipid which is PEG2000-DMG, or (c) The cationic lipid is HY-501, and the lipid nanoparticles further contain a helper lipid which is DSPC, or (d) The lipid nanoparticles i) 30 to 50 mol% of the cationic lipid, ii) 1-5 mol% PEG lipids, and iii) Contains 5-15 mol% of helper lipids, or (e) The lipid nanoparticles i) 30-50 mol% ALC-0315, ii) ALC-0159 in a concentration of 1-5% mol%, and iii) containing 5-15% mol% DSPC, or (f) The lipid nanoparticles i) 47.5 mol% ALC-0315, ii) 1.8 mol% ALC-0159, and iii) containing 10 mol% DSPC, or (g) The lipid nanoparticles i) SM-102 in a concentration of 30-50 mol%, ii) 1 to 5 mol% of PEG2000-DMG, and iii) containing 5 to 15 mol% DSPC, or (h) The lipid nanoparticles i) 50 mol% SM-102, ii) 1.5 mol% PEG2000-DMG and iii) A nucleic acid particle according to claim 6, comprising 10 mol% DSPC.

10. The aforementioned lipid nanoparticles (a) characterized by an N / P ratio of 4:1 to 12:1, and / or (b) characterized by an N / P ratio of 6:1, and / or (c) The nucleic acid particle according to claim 6, having a diameter of 50 nm to 150 nm as measured by dynamic laser light scattering (DLS).

11. The nucleic acid particle according to claim 1, wherein the nucleic acid particle comprises the immunomodulator, the cationic polymer, and RNA, and optionally the cationic polymer is poly(ethyleneimine) or poly(propyleneimine).

12. The nucleic acid particle according to claim 11, wherein the nucleic acid particle further comprises a secondary polymer, and optionally the secondary polymer is polysarcosine.

13. The nucleic acid particle according to claim 1, wherein the RNA is mRNA, and optionally the RNA is modRNA, circRNA, saRNA, taRNA, or uRNA.

14. Nucleic acid particles according to any one of claims 1 to 13, for use as a pharmaceutical.

15. A nucleic acid particle according to any one of claims 1 to 13, for use in the treatment and / or prevention of a disease or disorder, (a) The disease or disorder is an infectious disease, cancer, genetic disorder, autoimmune disease, or rare disease, and / or (b) The nucleic acid particles are administered parenterally or intranasally, and optionally intramuscularly, subcutaneously, intradermally, or intravenously.

Images