Cationic lipid compounds, preparation methods thereof, compositions and applications

Abstract

The present invention discloses a cationic lipid compound represented by formula (A), or an isomer thereof, or an N-oxide thereof, or a pharmaceutically acceptable salt or prodrug thereof. Also disclosed are a preparation method, a composition and an application of the above-mentioned cationic lipid compound. As a cationic lipid, such compounds are applicable to pharmaceutical compositions for nucleic acid delivery, and can achieve better stability, pKa value and intracellular delivery efficiency of drugs, wherein formula (A) is as follows.

Description

Technical Field

[0001] The present invention provides cationic lipids which can be used in combination with other lipid components (such as neutral lipids, steroids, and lipid-polymer conjugates) to form a nucleic acid mRNA lipid nanoparticle composition for delivering one or more therapeutic and / or prophylactic agents to mammalian cells or organs and / or for producing polypeptides in mammalian cells or organs. In addition to lipids, the lipid nanoparticle composition of the present invention may further include one or more cationic and / or ionizable amino lipids, neutral lipids including polyunsaturated lipids, lipid-polymer conjugates, steroids, and / or therapeutic and / or prophylactic agents in specific proportions. Background Art

[0002] The effective targeted delivery of bioactive substances such as small molecule drugs, proteins, and nucleic acids poses a persistent medical challenge. Specifically, delivering nucleic acids to cells is difficult due to the relative instability and low cellular permeability of these species. Therefore, there is a need to develop methods and compositions that facilitate the delivery of therapeutic and / or prophylactic agents such as nucleic acids to cells.

[0003] It has been demonstrated that lipid-based nanoparticle compositions, liposomes, and lipoplexes can be effectively used as delivery vehicles to transport bioactive substances such as small molecule drugs, proteins, and nucleic acids to cells and / or intracellular compartments. These compositions generally include one or more "cationic" lipids, neutral lipids (such as phospholipids) including polyunsaturated lipids, structural lipids (such as steroids), and / or lipids containing polyethylene glycol (lipid-polymer conjugates). Cationic lipids include amine-containing lipids that can be readily protonated.

[0004] However, the use of oligonucleotides in a therapeutic setting currently faces two problems. First, free RNA is readily digested by nucleases in plasma. Second, the ability of free RNA to enter intracellular compartments where the relevant translation machinery is present is limited. Lipid nanoparticles formed from cationic lipids and other lipid components (such as neutral lipids, cholesterol, PEG, PEGylated lipids, and oligonucleotides) have been used to prevent the degradation of RNA in plasma and to promote the cellular uptake of oligonucleotides.

[0005] There is still a need for improved cationic lipids and lipid nanoparticles for delivering oligonucleotides. Improved lipid nanoparticles would provide optimized drug delivery, protect nucleic acids from degradation and clearance in serum, be suitable for systemic or local delivery, and provide intracellular delivery of nucleic acids. Additionally, these preferred lipid-nucleic acid particles should be well-tolerated and provide a sufficient therapeutic index such that patient treatment at an effective dose of nucleic acid does not result in unacceptable toxicity and / or risk to the patient. The present invention provides these and related advantages. Summary of the Invention

[0006] The present invention provides the following compounds and methods related to these compounds:

[0007] In a first aspect, the present invention relates to a compound represented by formula (I), or an isomer thereof, or an N-oxide thereof, or a pharmaceutically acceptable salt, prodrug thereof;

[0008] Wherein:

[0009]

[0010] L1 is selected from the following structures: -C-, -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O) X -, -S-S-, -C(=O)S-, -SC(=O)-, -N-C(=O)-, -C(=O)-N-;

[0011] R1 and R2 are each independently a C6-C24 alkyl or C6-C24 alkenyl, and the hydrocarbon chain is optionally connected by one or more ester bonds or ether bonds;

[0012] R3 and R4 are each independently a C1-C12 alkyl or C1-C12 alkenyl, or R3 and R4 combine with each other to form a 4- to 10-membered heterocycle, and the heteroatoms include one or more heteroatoms selected from N, O, S, and the heterocycle is optionally substituted by 1-6 heteroatoms;

[0013] X is C, N, O, S, -S-S-;

[0014] M is a C1-C12 alkyl or C1-C12 alkenyl;

[0015] x is 0, 1 or 2.

[0016] In various different embodiments, the compound has one of the structures shown in Table 1 below

[0017] Table 1 Representative Compounds

[0018]

[0019]

[0020]

[0021]

[0022]

[0023]

[0024]

[0025]

[0026]

[0027] In some embodiments, there are provided compositions comprising any one or more of the compounds of structural formula (I) and a therapeutic agent and / or a prophylactic agent.

[0028] In some embodiments, there are provided compositions comprising any one or more of the compounds of structure (I) and a therapeutic agent and / or a prophylactic agent. In some embodiments, the composition comprises any one of the compounds of structure (I) and a therapeutic agent and / or a prophylactic agent, and one or more excipients selected from neutral lipids, steroids, and polymer-conjugated lipids. Other pharmaceutically acceptable excipients and / or carriers are also included in various embodiments of the composition.

[0029] In some embodiments, the neutral lipids are selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), sphingomyelin (SM), and mixtures thereof. In some embodiments, the preferred neutral lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).

[0030] In some embodiments, the steroids are selected from cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof. In some embodiments, the preferred steroid is cholesterol.

[0031] In some embodiments, the polyethylene glycolylated lipid is 1,2-dimyristoyl-sn-glycero methoxypolyethylene glycol (PEG-DMG)

[0032] In some embodiments, the composition is in the following proportion ranges: about 10-60 mol% of the compound, about 0-30 mol% of neutral lipid, about 10-55 mol% of steroid, and about 0-10 mol% of polymer-conjugated lipid.

[0033] In some embodiments of the foregoing compositions, the therapeutic and / or prophylactic agent comprises a nucleic acid. Wherein the nucleic acid is RNA, which is selected from the group consisting of siRNA, aiRNA, miRNA, dsRNA, shRNA, mRNA, and mixtures thereof. In some embodiments, the RNA is selected from mRNA.

[0034] In other different embodiments, the present invention relates to a method of administering a therapeutic and / or prophylactic agent to a subject in need thereof, the method comprising preparing or providing any one of the above compositions and administering the composition to the subject.

[0035] For the purpose of administration, the compounds of the present invention (usually in the form of lipid nanoparticles combined with a therapeutic and / or prophylactic agent) can be administered as the active pharmaceutical ingredient, or can be formulated into a pharmaceutical composition. The pharmaceutical composition of the present invention comprises a compound of formula (I) and one or more pharmaceutically acceptable carriers, diluents or excipients. The compound of formula (I) is effective to form lipid nanoparticles and deliver the therapeutic and / or prophylactic agent. Those skilled in the art can readily determine the appropriate concentration and dosage.

[0036] The administration of the compositions of the present invention can be carried out by any acceptable mode of administration for reagents of similar utility. The pharmaceutical compositions of the present invention can be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols. Typical routes of administration of such pharmaceutical compositions include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal and intranasal routes. The term parenteral as used herein includes subcutaneous injection, intravenous, intramuscular, intradermal, intrasternal injection or infusion techniques. The pharmaceutical compositions of the present invention are formulated to allow the active ingredient contained therein to be bioavailable after administration of the composition to the subject. The composition to be administered to the subject or patient is in the form of one or more dosage units, wherein a tablet can be a single dosage unit, and a container of the compound in aerosol form of the present invention can contain multiple dosage units. Current methods of preparing these dosage forms are known or will be apparent to those skilled in the art. In any case, the composition to be administered will contain a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof to treat the relevant disease or condition according to the teachings of the present invention.

[0037] The pharmaceutical compositions of the present invention can be in solid or liquid form. In one aspect, the carrier is a microparticle such that the composition is in the form of a tablet or powder. The carrier can be a liquid, in which case the composition is an oral syrup or an injectable liquid or an aerosol suitable for inhalation administration.

[0038] When intended for oral administration, the pharmaceutical composition is preferably in solid or liquid form, where forms considered solid or liquid herein include semi-solid, semi-liquid, suspension and gel forms.

[0039] As a solid composition for oral administration, the pharmaceutical composition can be formulated in the form of powders, granules, compressed tablets, pills, capsules, chewing gums, wafers, etc. Such solid compositions will generally contain one or more inert diluents or edible carriers. Additionally, one or more of the following may be present: binders such as gelatin, cellulose, etc.; excipients such as lactose, etc.; disintegrants such as alginic acid, etc.; lubricants such as magnesium stearate, etc.; glidants such as silica gel, etc.; sweeteners such as sucrose or saccharin; flavoring agents such as peppermint, etc.; and coloring agents.

[0040] When the pharmaceutical composition is in capsule form, it can contain a liquid carrier in addition to the materials of the above types, such as polyethylene glycol or oil.

[0041] The pharmaceutical composition can be in liquid form, such as syrups, solutions, emulsions or suspensions. As two examples, the liquid can be for oral administration or for injectable delivery. When intended for oral administration, the preferred composition contains, in addition to the compound of the present invention, one or more of sweeteners, preservatives, coloring / coloring agents and flavoring agents. In compositions administered by injection, one or more of surfactants, preservatives, wetting agents, dispersing agents, suspending agents, buffering agents, stabilizers and isotonic agents can be included.

[0042] The liquid pharmaceutical compositions of the present invention, whether in solution, suspension or other similar forms, can include one or more of the following adjuvants: sterile diluents such as water for injection, saline solutions, preferably physiological saline, Ringer's solution, isotonic sodium chloride; non-volatile oils such as synthetic monoglycerides or diglycerides, polyethylene glycol, glycerol, propylene glycol or other solvents that can be used as solvents or suspending media; antibacterial agents such as methylparaben, etc.; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffering agents such as acetate, citrate or phosphate; and reagents for adjusting tonicity such as sodium chloride or glucose; reagents used as cryoprotectants such as sucrose or trehalose. Parenteral preparations can be enclosed in ampoules, disposable syringes or multi-dose vials made of glass or plastic. Physiological saline is the preferred adjuvant. Injectable pharmaceutical compositions are preferably sterile.

[0043] The pharmaceutical composition of the present invention may consist of dosage units that can be administered as an aerosol. The term aerosol is used to denote a variety of systems ranging from colloidal systems to systems consisting of pressurized packages. It can be delivered by liquefied gas or compressed gas, or by a suitable pump system that disperses the active ingredient. The aerosol of the compounds of the present invention can be delivered in a single-phase, two-phase system or a three-phase system in order to deliver the active ingredient. The delivery of the aerosol includes the necessary containers, activators, valves, sub-containers, etc., which together can form a kit. A person skilled in the art can determine the preferred aerosol without undue experimentation.

[0044] The pharmaceutical composition of the present invention can be prepared by methods well known in the pharmaceutical art. The pharmaceutical composition intended to be administered by injection can be prepared by combining the lipid nanoparticles of the present invention with sterile distilled water or other carriers to form a solution. Surfactants can be added to facilitate the formation of a uniform solution or suspension. A surfactant is a compound that interacts non-covalently with the compounds of the present invention in order to facilitate the dissolution or uniform suspension of the compound in an aqueous delivery system.

[0045] The composition of the present invention or a pharmaceutically acceptable salt thereof is administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of the specific therapeutic agent used; the metabolic stability and duration of action of the therapeutic agent; the age, weight, general health, sex and diet of the subject; the mode and time of administration; the excretion rate; drug combinations; the severity of the specific case, etc.

[0046] The composition of the present invention can also be administered simultaneously with, before or after the administration of one or more other therapeutic agents. Such combination therapies include single pharmaceutical dosage formulations that administer the composition of the present invention and one or more additional active agents, as well as the administration of the composition of the present invention and the active agents in their own separate pharmaceutical dosage formulations. For example, the composition of the present invention and other active agents can be administered to a subject together in a single oral dosage composition (such as a tablet or capsule), or each reagent can be administered in different oral dosage formulations. When using different dosage formulations, the compounds of the present invention and one or more additional active agents can be administered at substantially the same time, or sequentially at staggered times; it should be understood that combination therapy includes all of these dosing regimens.

[0047] The structural modification and design of the above-mentioned cationic lipid compounds have achieved more advantageous physicochemical properties, including a more suitable pKa and better chemical stability, for mRNA nano-liposome compositions, which can achieve more effective binding and delivery of ionic nucleic acid drugs. At the same time, its chemical structure is more stable, facilitating synthesis and being favorable for development as a pharmaceutical excipient.

[0048] The preparation methods of the above-mentioned compounds and compositions are described below and / or are known in the art.

[0049] Those skilled in the art will recognize that in the methods described herein, the functional groups of the intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include hydroxyl, amino, and carboxylic acid. Suitable protecting groups for hydroxyl include trialkylsilyl or diarylalkylsilyl, tetrahydrofuranyl, benzyl, etc. Suitable protecting groups for amino include tert-butoxycarbonyl, benzyloxycarbonyl, etc. Suitable protecting groups for carboxylic acid include alkyl, aryl, or aralkyl esters. The protecting groups can be added or removed according to standard techniques, which are known to those skilled in the art and described herein.

[0050] Those skilled in the art will also recognize that although such protected derivatives of the compounds of the present invention may not thereby have pharmaceutical activity, they can be administered to mammals and then metabolized in vivo to form the pharmaceutically active compounds of the present invention. Such derivatives can therefore be described as "prodrugs". Thus, prodrugs of the compounds of the present invention are included within the scope of the present invention.

[0051] In addition, all compounds of the present invention in the form of free bases or free acids can be converted into their pharmaceutically acceptable salts by treatment with appropriate inorganic or organic bases or acids according to methods known to those skilled in the art. The salts of the compounds of the present invention can be converted into their free base or acid forms by standard techniques.

[0052] The following examples are provided for purposes of illustration and not limitation.

[0053] In the following examples, unless otherwise indicated, all solvents and reagents used are commercially available and used as received.

[0054] The following abbreviations are used herein:

[0055] SM1: Diethyl malonate

[0056] SM2: 1-Bromononane

[0057] KOH: Potassium hydroxide

[0058] LiBH4: Lithium borohydride

[0059] EtOH: Ethanol

[0060] SM4: 2-Octyldecanoic acid. Detailed Description

[0061] Example 1:

[0062] Representative Route

[0063]

[0064] Synthesis of Compound 1: 2-((2-(Diethylamino)ethyl)formamido)undecyl Hexyl 2-Decanoate

[0065] 1) Synthesis chemical formula of Compound A: C 16 H 30 O4 Molecular weight: 286.41

[0066] Add anhydrous ethanol (200 ml) into a 500 ml three-necked flask. Add sodium metal (11.5 g) under an ice-water bath and stir until completely dissolved. Dropwise add diethyl malonate (80.9 g, 0.5 mol) and 1-bromononane (103.6 g, 0.5 mol) in sequence, and reflux for 2 h. A white solid precipitates during the stirring process. Cool down and filter. Concentrate the filtrate and replace it with an ethyl acetate (200 ml) solution. Wash it once with 100 ml of water, and back-extract the aqueous phase twice with ethyl acetate (100 ml * 2). After combining the organic phases, dry the organic phase with 10 g of anhydrous magnesium sulfate, and concentrate to dryness to obtain a colorless oily liquid Compound A: 141.0 g, with a yield of 98%.

[0067] LC-MS (ESI, m / z, C 16 H 30 O4, 287.2, M+H)

[0068] HNMR (DMSO-d6, 400 MHz) δ 4.14 (m, 4H), 3.31 (m, 1H), 1.78 (m, 2H), 1.40 (m, 2H), 1.26 - 1.21 (m, 18H), 0.88 (m, 3H).

[0069] 2) Synthesis of Compound B

[0070]

[0071] Chemical formula: C 12 H 24 O3

[0072] Molecular weight: 216.32

[0073] Add Compound A (114.6 g, 0.4 mol) and ethanol (500 mL) into a 1 L three-necked flask. Dropwise add a solution of KOH (22.5 g, 0.4 mol) in ethanol (250 ml). Stir at room temperature overnight. Concentrate to remove the solvent. Dilute the resulting emulsion with 150 ml of water and 50 ml of saturated sodium bicarbonate aqueous solution, wash with ethyl acetate. Adjust the pH of the aqueous phase to below 2 with sulfuric acid, then extract with ethyl acetate. Combine the ethyl acetate phases, dry with anhydrous magnesium sulfate, and concentrate to dryness to obtain a colorless oily compound: 98.2 g, with a yield of 95%. Without purification, it is directly used in the next step of the reaction.

[0074] Add lithium borohydride (43.6 g, 2 mol) and tetrahydrofuran (700 ml) into a 3 L three-necked reaction flask, stir, and while controlling the temperature with an ice-water bath, dropwise add an isopropanol solution (800 ml) of the oily substance from the previous step (98.2 g, 0.38 mol). After addition, react at room temperature for 3 h. Adjust the pH ≤ 1 with hydrochloric acid under an ice-water bath, filter, replace the filtrate with an ethyl acetate (500 ml) solution, wash once with 200 ml of water, and back-extract the aqueous phase twice with ethyl acetate (200 ml * 2). After combining the organic phases, dry the organic phase with 30 g of anhydrous magnesium sulfate, and concentrate to dryness to obtain a pale yellow oily liquid compound B: 82.2 g, with a yield of 95%.

[0075] LC-MS (ESI, m / z, C 12 H 24 O3, 217.2, M + H)

[0076] 1H NMR (DMSO-d6, 400 MHz) δ 10.63 (s, 1H), 6.34 (s, 1H), 3.87 - 3.61 (m, 2H), 2.28 - 2.26 (m, 1H), 1.50 - 1.47 (m, 2H), 1.29 - 1.25 (m, 14H), 0.88 (t, J = 8.0 Hz, 3H).

[0077] 3) Synthesis of compound C

[0078]

[0079] Chemical formula: C 18 H 38 N2O2

[0080] Molecular weight: 314.51

[0081] Add compound B (65.0 g, 0.3 mol), N,N-diethylethylenediamine (38.3 g, 0.33 mol) and water (1 L) into a 3 L three-necked reaction flask. Stir and control the temperature at 5 - 15 °C. At this temperature, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (69.0 g, 0.36 mol) in portions. After addition, stir at room temperature for 2 h. Filter, wash the filter cake with petroleum ether, and dry in vacuo at 40 - 50 °C to obtain white solid C: 75.5 g, with a yield of 80%.

[0082] LC-MS (ESI, m / z, C 18 H 38 N2O2, 315.3, M + H)

[0083] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 6.34 (s, 1H), 3.88 - 3.62 (m, 2H), 3.30 (m, 2H), 2.46 - 2.34 (m, 7H), 1.49 (m, 2H), 1.26 - 1.25 (m, 14H), 0.93 - 0.88 (m, 9H).

[0084] 4) Synthesis of Compound 1

[0085]

[0086] Chemical formula: C 34 H 68 N2O3

[0087] Molecular weight: 552.93

[0088] Add 2-hexyldecanoic acid (51.3 g, 0.2 mol), toluene (400 ml) and DMF (2 ml) into a 1 L three-necked flask. Oxalyl chloride was added dropwise with stirring. After the addition, the mixture was stirred at room temperature for 2 hours and then concentrated to obtain the acyl chloride intermediate.

[0089] Add Compound C (62.9 g, 0.2 mol), TEA (60.7 g, 0.6 mol) and tetrahydrofuran (400 mL) into a 2 L three-necked flask. The acyl chloride intermediate was added dropwise with stirring at room temperature. After the addition, the mixture was stirred at 40 - 50 °C for 2 hours. After monitoring the reaction by TLC and determining that it was basically complete, ethyl acetate was added. The organic phase was washed successively with water, hydrochloric acid, sodium bicarbonate and water, transferred to a single-necked flask, silica gel was added, and the mixture was directly concentrated under reduced pressure and stirred for sample preparation. Purification by column chromatography (DCM:MeOH = 100:5 - 100:10, adding 1% ammonia water) gave Compound 1 (75.2 g), a pale yellow oil, with a yield of 68%.

[0090] LC-MS (ESI, m / z, C 34 H 68 N2O3, 553.5, M+H)

[0091] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 (m, 6H), 2.13 (m, 1H), 1.60 - 1.49 (m, 6H), 1.29 - 1.26 (m, 38H), 0.93 - 0.88 (m, 15H).

[0092] Examples 2 - 56 were prepared by methods similar to those described in Example 1 or by methods known in the art.

[0093] Example 2

[0094] Synthesis of 6-bromohexyl 2-octyldecanoate

[0095]

[0096] 6-bromohexyl 2-hexyldecanoate

[0097] Chemical formula: C 22 H 43 BrO2

[0098] Molecular weight: 419.49

[0099] Into a 500 ml three-necked reaction flask, 2-hexyldecanoic acid (10 g, 39.0 mmol) and tetrahydrofuran (200 ml) were added, and 6-bromohexanol (9.18 g, 50.7 mmol) and potassium carbonate (7.0 g, 50.7 mmol) were added successively. The temperature was adjusted to 60 - 70 °C, and the mixture was stirred and reacted for 8 h. After the reaction was detected to be complete by in-process control, it was cooled to room temperature. It was distilled under reduced pressure, water and ethyl acetate were added, and after liquid separation, it was extracted with ethyl acetate. After combining the organic phases, it was washed with an aqueous sodium bicarbonate solution and a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and distilled under reduced pressure to obtain the crude product. Column chromatography (eluted with petroleum ether:ethyl acetate = 10:1) gave 6-bromohexyl 2-octyldecanoate: 13.1 g, a yellow oil, with a yield of 80%.

[0100] LC-MS (ESI, m / z, C 22 H 43 BrO2, 419.2, M+H)

[0101] 1H NMR (DMSO-d6, 400 MHz) δ 4.06 (m, 2H), 3.52 (m, 2H), 2.13 (m, 1H), 1.82 (m, 2H), 1.60 (m, 6H), 1.43 (m, 2H), 1.29 - 1.25 (m, 22H), 0.88 (m, 6H).

[0102] Example 3

[0103] Synthesis of 6-(pentadecyl 7-oxy)-6-oxohexanoic acid

[0104]

[0105] 6-(pentadecyl 7-oxy)-6-oxohexanoic acid

[0106] Chemical formula: C 21 H 40 O4

[0107] Molecular weight: 356.55

[0108] Into a 500 ml three-necked reaction flask, add 7-pentadecanol (10 g, 43.8 mmol) and N,N-dimethylformamide (100 ml), and successively add adipic acid (6.4 g, 43.8 mmol) and sodium hydroxide (2.6 g, 65.7 mmol). Adjust the temperature to 70 - 80 °C and stir the reaction for 6 h. After monitoring the reaction to completion by in-process control, cool it to room temperature. Perform vacuum distillation, add water and ethyl acetate, extract with ethyl acetate after liquid separation, combine the organic phases, wash with an aqueous sodium bicarbonate solution and a saturated brine solution, dry over anhydrous sodium sulfate, and perform vacuum distillation to obtain the crude product. Column chromatography (elution with petroleum ether:ethyl acetate = 1:1) gives 6-(heptadecyl 9-oxy)-6-oxohexanoic acid: 9.4 g, a yellow oil, with a yield of 60.0%.

[0109] LC-MS(ESI, m / z, C 21 H 40 O4, 357.3, M+H)

[0110] 1H NMR(DMSO-d6, 400 MHz) δ 4.47(m, 1H), 2.32(m, 2H), 2.21(m, 2H), 1.64(m, 2H), 1.52(m, 2H), 1.49(m, 4H), 1.26(m, 16H), 0.90(m, 6H).

[0111] Example 4

[0112] Synthesis of Compound 6-bromohexane-7-pentadecyl ether

[0113]

[0114] Compound: 6-bromohexane-7-pentadecyl ether

[0115] Chemical formula: C13H 43 BrO

[0116] Molecular weight 391.48

[0117] Into a 500 ml three-necked reaction flask, add 7-pentadecanol (10 g, 43.8 mmol) and N,N-dimethylformamide (100 ml). Sequentially add TBAB (1.12 g, 3.5 mmol), sodium hydroxide (50% aqueous solution, 5.3 g, 65.7 mmol) and 1,6-dibromohexane (10.7 g, 43.8 mmol). Adjust the temperature to 50 - 60 °C and stir the reaction for 8 h. After detecting the completion of the reaction by in-process control, cool to room temperature. Perform vacuum distillation, add water and ethyl acetate, extract with ethyl acetate after phase separation. After combining the organic phases, wash with aqueous sodium bicarbonate solution and saturated brine solution, dry over anhydrous sodium sulfate, and perform vacuum distillation to obtain the crude product. Subject to column chromatography (eluting with petroleum ether:ethyl acetate = 1:1) to obtain 6-bromohexyl-7-pentadecyl ether: 12.0 g, a yellow oil, with a yield of 70.0%.

[0118] LC-MS(ESI, m / z, C 23 H 47 BrO, 391.3, M+H)

[0119] 1H NMR(DMSO-d6, 400 MHz) δ 3.52 - 3.35(m, 4H), 3.10(m, 1H), 1.82(m, 2H), 1.50 - 1.38(m, 8H), 1.29 - 1.26(m, 22H), 0.88(m, 6H).

[0120] Example 5

[0121] Synthesis of Compound 2

[0122]

[0123] Chemical formula: C 34 H 68 N2O3

[0124] Molecular weight: 552.93

[0125] Refer to the synthesis in Example 1.

[0126] LC-MS(ESI, m / z, C 34 H 68 N2O3, 553.5, M+H)

[0127] 1H NMR(DMSO-d6, 400 MHz) δ 8.01(s, 1H), 4.42 - 4.17(m, 2H), 3.30(m, 2H), 2.93(m, 1H), 2.46 - 2.32(m, 8H), 1.66 - 1.49(m, 4H), 1.33 - 1.25(m, 38H), 0.93 - 0.88(m, 12H). A pale yellow oil.

[0128] Example 6

[0129] Synthesis of Compound 3

[0130]

[0131] Chemical formula: C 31 H 60 N2O3

[0132] Molecular weight: 508.83

[0133] Refer to the synthesis in Example 1.

[0134] LC-MS (ESI, m / z, C 31 H 60 N2O3, 509.5, M+H)

[0135] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 - 2.32 (m, 8H), 1.66 - 1.49 (m, 8H), 1.37 - 1.26 (m, 32H), 0.88 (m, 6H). Pale yellow oil.

[0136] Example 7

[0137] Synthesis of Compound 4

[0138]

[0139] Chemical formula: C 40 H 78 N2O4

[0140] Molecular weight: 651.07

[0141] Refer to the synthesis in Example 1.

[0142] LC-MS (ESI, m / z, C 40 H 78 N2O4, 651.6, M+H)

[0143] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 4.49 - 4.17 (m, 3H), 3.60 (m, 1H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 - 2.32 (m, 8H), 1.77 - 1.49 (m, 8H), 1.33 - 1.25 (m, 48H), 0.88 (m, 6H). Pale yellow oil.

[0144] Example 8

[0145] Synthesis of Compound 5

[0146]

[0147] Chemical formula: C 39 H 76 N2O4

[0148] Molecular weight: 637.05

[0149] See the synthesis in Example 1.

[0150] LC-MS (ESI, m / z, C 39 H 76 N2O4, 637.6, M+H)

[0151] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.52 (m, 4H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 - 2.32 (m, 8H), 1.66 - 1.49 (m, 4H), 1.33 - 1.25 (m, 48H), 0.88 (m, 6H). Pale yellow oil.

[0152] Example 9

[0153] Synthesis of Compound 6

[0154]

[0155] Chemical formula: C 40 H 79 N3O3

[0156] Molecular weight: 650.09

[0157] See the synthesis in Example 1.

[0158] LC-MS (ESI, m / z, C 40 H 79 N3O3, 650.6, M+H)

[0159] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 - 2.29 (m, 12H), 2.14 (s, 3H), 1.66 - 1.49 (m, 4H), 1.33 - 1.25 (m, 48H), 0.88 (m, 6H). Pale yellow oil.

[0160] Example 10

[0161] Synthesis of Compound 7

[0162] Chemical formula: C 33 H 64 N2O3

[0163]

[0164] Molecular weight: 536.89

[0165] See the synthesis in Example 1.

[0166] LC-MS (ESI, m / z, C 33 H 64 N2O3, 537.5, M+H)

[0167] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.30 (m, 2H), 2.93 (m, 1H), 2.51 - 2.32 (m, 8H), 1.68 - 1.49 (m, 8H), 1.33 - 1.25 (m, 36H), 0.88 (m, 6H). Pale yellow oil.

[0168] Example 11

[0169] Synthesis of Compound 8

[0170]

[0171] Chemical formula: C 39 H 78 N2O5

[0172] Molecular weight: 655.06

[0173] See the synthesis in Example 1.

[0174] LC-MS (ESI, m / z, C 39 H 78 N2O5, 655.6, M+H)

[0175] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.16 (m, 4H), 3.42 - 3.30 (m, 6H), 2.93 (m, 1H), 2.57 - 2.32 (m, 8H), 1.66 - 1.49 (m, 4H), 1.33 - 1.25 (m, 48H), 0.88 (m, 6H). Pale yellow oil.

[0176] Example 12

[0177] Synthesis of Compound 9

[0178]

[0179] Chemical formula: C 36 H 72 N2O3

[0180] Molecular weight: 580.98

[0181] See the synthesis in Example 1.

[0182] LC-MS (ESI, m / z, C 36 H 72 N2O3, 581.6, M+H)

[0183] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 (m, 6H), 2.13 (s, 1H) 1.60 - 1.49 (m, 6H), 1.31 - 1.25 (m, 38H), 0.93 - 0.88 (m, 15H). Pale yellow oil.

[0184] Example 13

[0185] Synthesis of Compound 10

[0186] Chemical formula: C 45 H 88 N2O5

[0187]

[0188] Molecular weight: 737.21

[0189] See the syntheses in Example 1 and Example 2.

[0190] LC-MS (ESI, m / z, C 47 H 92 N2O5, 737.7, M+H)

[0191] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.13 (m, 4H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 (m, 6H), 2.32 (m, 2H), 2.13 (m, 1H), 1.60 - 1.43 (m, 12H), 1.33 - 1.25 (m, 44H), 0.93 - 0.88 (m, 15H). Pale yellow oil.

[0192] Example 14

[0193] Synthesis of Compound 11

[0194] Chemical formula: C 47 H 90 N2O5

[0195]

[0196] Molecular weight: 763.25

[0197] See the synthesis in Examples 1 and 2.

[0198] LC-MS (ESI, m / z, C 47 H 90 N2O5, 763.7, M+H)

[0199] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.13 (m, 4H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 - 2.42 (m, 6H), 2.32 (m, 2H), 2.13 (m, 1H), 1.66 - 1.60 (m, 8H), 1.49 - 1.25 (m, 56H), 0.88 (m, 9H). Pale yellow oil.

[0200] Example 15

[0201] Synthesis of Compound 12

[0202]

[0203] Chemical formula: C 47 H 90 N2O6

[0204] Molecular weight: 779.25

[0205] See the synthesis in Examples 1 and 2.

[0206] LC-MS (ESI, m / z, C 47 H 90 N2O6, 779.7, M+H)

[0207] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.49 (m, 1H), 4.42 - 4.13 (m, 4H), 3.60 (m, 1H), 3.30 (m, 2H), 2.93 (m, 1H), 2.51 - 2.32 (m, 8H), 2.13 (m, 1H), 1.77 - 1.43 (m, 16H), 1.33 - 1.25 (m, 46H), 0.88 (m, 9H)

[0208] Example 16

[0209] Synthesis of Compound 13

[0210]

[0211] Chemical formula: C 46 H 88 N2O6

[0212] Molecular weight: 765.22

[0213] See the synthesis in Example 1 and Example 2.

[0214] LC-MS (ESI, m / z, C 46 H 88 N2O6, 765.7, M+H)

[0215] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.13 (m, 4H), 3.52 (t, 4H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 - 2.32 (m, 8H), 2.13 (m, 1H), 1.66 - 1.43 (m, 12H), 1.33 - 1.26 (m, 46H), 0.88 (m, 9H). Pale yellow oil.

[0216] Example 17

[0217] Synthesis of Compound 14

[0218]

[0219] Chemical formula: C 47 H 91 N3O5

[0220] Molecular weight: 778.26

[0221] See the synthesis in Example 1 and Example 2.

[0222] LC-MS (ESI, m / z, C 47 H 91 N3O5, 778.7, M+H)

[0223] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.13 (m, 4H), 3.52 (t, 4H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 - 2.32 (m, 8H), 2.14 - 2.13 (m, 4H), 1.66 - 1.43 (m, 12H), 1.33 - 1.26 (m, 46H), 0.88 (m, 9H)

[0224] Example 18

[0225] Synthesis of Compound 15

[0226]

[0227] Chemical formula: C 46 H 90 N2O7

[0228] Molecular weight: 783.23

[0229] Refer to the synthesis in Example 1 and Example 2.

[0230] LC-MS (ESI, m / z, C 46 H 90 N2O7, 783.7, M+H)

[0231] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.13 (m, 6H), 3.42 (t, 4H), 3.30 (m, 2H), 2.93 (m, 1H), 2.57 - 2.32 (m, 8H), 2.13 (m, 1H), 1.66 - 1.43 (m, 12H), 1.33 - 1.26 (m, 46H), 0.88 (m, 9H). Pale yellow oil.

[0232] Example 19

[0233] Synthesis of Compound 16

[0234] Chemical formula: C 47 H 92 N2O5

[0235] Molecular weight: 765.26

[0236]

[0237] Refer to the synthesis in Example 1 and Example 4.

[0238] LC-MS (ESI, m / z, C 47 H 92 N2O5, 765.7, M+H)

[0239] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.13 (m, 4H), 3.30 (m, 2H), 2.93 (m, 1H), 2.46 - 2.32 (m, 8H), 2.13 (m, 1H), 1.66 - 1.49 (m, 10H), 1.33 - 1.25 (m, 50H), 0.93 - 0.88 (m, 15H). Pale yellow oil.

[0240] Example 20

[0241] Synthesis of Compound 17

[0242]

[0243] Chemical formula: C 48 H 94 N2O4

[0244] Molecular weight: 763.29

[0245] See the synthesis in Example 1 and Example 4.

[0246] LC-MS (ESI, m / z, C 48 H 94 N2O4, 763.7, M+H)

[0247] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.13 (m, 2H), 3.35 - 3.30 (m, 4H), 3.10 - 2.93 (m, 2H), 2.46 - 2.32 (m, 8H), 1.66 - 1.43 (m, 12H), 1.38 - 1.25 (m, 56H), 0.88 (m, 9H). Pale yellow oil.

[0248] Example 21

[0249] Synthesis of Compound 18

[0250]

[0251] Chemical formula: C 47 H 92 N2O5

[0252] Molecular weight: 765.26

[0253] See the synthesis in Example 1 and Example 4.

[0254] LC-MS (ESI, m / z, C 47 H 92 N2O5, 765.7, M+H)

[0255] HNMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.52 (m, 4H), 3.35 - 3.30 (m, 4H), 3.10 - 2.93 (m, 2H), 2.46 - 2.32 (m, 8H), 1.66 - 1.43 (m, 8H), 1.38 - 1.25 (m, 54H), 0.88 (m, 9H). Pale yellow oil.

[0256] Example 22

[0257] Synthesis of Compound 19

[0258]

[0259] Chemical formula: C 48 H 95 N3O4

[0260] Molecular weight: 778.31

[0261] See the synthesis in Example 1 and Example 4.

[0262] LC-MS (ESI, m / z, C 48 H 95 N3O4, 778.7, M + H)

[0263] HNMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.35 - 3.30 (m, 4H), 3.10 - 2.93 (m, 2H), 2.46 - 2.29 (m, 12H), 2.14 (s, 3H) 1.66 - 1.43 (m, 8H), 1.38 - 1.25 (m, 54H), 0.88 (m, 9H). Pale yellow oil.

[0264] Example 23

[0265] Synthesis of Compound 20

[0266]

[0267] Chemical formula: C 47 H 94 N2O6

[0268] Molecular weight: 783.28

[0269] See the synthesis in Example 1 and Example 4.

[0270] LC-MS (ESI, m / z, C 47 H 94 N2O6, 783.7, M + H)

[0271] HNMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.16 (m, 4H), 3.35 - 3.30 (m, 8H), 3.10 - 2.93 (m, 2H), 2.57 - 2.32 (m, 8H), 1.66 - 1.43 (m, 8H), 1.38 - 1.25 (m, 54H), 0.88 (m, 9H). Pale yellow oil.

[0272] Example 24

[0273] Synthesis of Compound 21

[0274]

[0275] Chemical formula: C 52 H 104 N2O5

[0276] Molecular weight: 837.41

[0277] See the synthesis in Example 1, Example 2 and Example 4.

[0278] LC-MS (ESI, m / z, C 52 H 104 N2O5, 837.8, M+H)

[0279] HNMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.13 (m, 2H), 3.71 - 3.30 (m, 8H), 2.54 - 2.46 (m, 7H), 2.13 (m, 1H), 1.60 - 1.38 (m, 23H), 1.29 - 1.25 (m, 44H), 0.93 - 0.88 (m, 18H). Pale yellow oil.

[0280] Example 25

[0281] Synthesis of Compound 22

[0282] Chemical formula: C 53 H 104 N2O5

[0283]

[0284] Molecular weight 849.42

[0285] See the synthesis in Example 1, Example 2 and Example 4.

[0286] LC-MS (ESI, m / z, C 53 H 104 N2O5, 849.8, M+H)

[0287] HNMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.13 (m, 2H), 3.71 - 3.30 (m, 8H), 3.10 (m, 1H), 2.54 - 2.42 (m, 7H), 2.13 (m, 1H), 1.60 - 1.38 (m, 16H), 1.29 - 1.25 (m, 50H), 0.93 - 0.88 (m, 18H). Pale yellow oil.

[0288] Example 26

[0289] Synthesis of Compound 23

[0290]

[0291] Chemical formula: C 53 H 104 N2O6

[0292] Molecular weight: 865.42

[0293] See the synthesis in Example 1, Example 2 and Example 4.

[0294] LC-MS (ESI, m / z, C 53 H 104 N2O6, 865.8, M + H)

[0295] HNMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.49 (s, 1H), 4.13 (m, 2H), 3.71 - 3.30 (m, 9H), 3.10 (m, 1H), 2.54 - 2.41 (m, 7H), 2.13 (m, 1H), 1.60 - 1.38 (m, 26H), 1.29 - 1.25 (m, 44H), 0.93 - 0.88 (m, 12H). Pale yellow oil.

[0296] Example 27

[0297] Synthesis of Compound 24

[0298] Chemical formula: C 52 H 102 N2O6

[0299]

[0300] Molecular weight: 851.40

[0301] See the synthesis in Example 1, Example 2 and Example 4.

[0302] LC-MS (ESI, m / z, C 52 H102 N2O6, 851.8, (M+H)

[0303] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.13 (m, 2H), 3.71 - 3.30 (m, 12H), 3.10 (m, 1H), 2.54 - 2.40 (m, 7H), 2.13 (m, 1H), 1.60 - 1.38 (m, 22H), 1.29 - 1.25 (m, 44H), 0.93 - 0.88 (m, 12H). Pale yellow oil.

[0304] Example 28

[0305] Synthesis of Compound 25

[0306] Chemical formula: C 53 H 105 N3O5

[0307]

[0308] Molecular weight: 864.44

[0309] See the synthesis in Example 1, Example 2 and Example 4.

[0310] LC-MS (ESI, m / z, C 53 H 105 N3O5, 864.8, (M+H)

[0311] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.13 (m, 2H), 3.71 - 3.30 (m, 12H), 3.10 (m, 1H), 2.54 - 2.40 (m, 7H), 2.14 - 2.13 (m, 4H), 1.60 - 1.38 (m, 22H), 1.29 - 1.25 (m, 52H), 0.93 - 0.88 (m, 12H). Pale yellow oil.

[0312] Example 29

[0313] Synthesis of Compound 26

[0314] Chemical formula: C 52 H 102 N2O5

[0315]

[0316] Molecular weight: 835.4

[0317] See the synthesis in Example 1, Example 2 and Example 4.

[0318] LC-MS (ESI, m / z, C 52 H 108 N2O5, 835.8, M+H)

[0319] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 4.13 (m, 2H), 3.71 - 3.30 (m, 8H), 3.10 (m, 1H), 2.54 - 2.42 (m, 7H), 2.13 (m, 1H), 1.68 - 1.38 (m, 26H), 1.29 - 1.25 (m, 44H), 0.93 - 0.88 (m, 12H). Pale yellow oil.

[0320] Example 30

[0321] Synthesis of Compound 27

[0322]

[0323] Chemical formula: C 52 H 104 N2O7

[0324] Molecular weight: 869.41

[0325] See the synthesis in Example 1, Example 2 and Example 4.

[0326] LC-MS (ESI, m / z, C 52 H 104 N2O7, 869.8, M+H)

[0327] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 4.16 - 4.13 (m, 4H), 3.71 - 3.35 (m, 12H), 3.10 (m, 1H), 2.57 - 2.46 (m, 7H), 2.13 (m, 1H), 1.60 - 1.38 (m, 22H), 1.29 - 1.26 (m, 44H), 0.93 - 0.88 (m, 12H). Pale yellow oil.

[0328] Example 31

[0329] Synthesis of Compound 28

[0330] Chemical formula: C 50 H 100 N2O5

[0331]

[0332] Molecular weight: 809.36

[0333] See the synthesis in Example 1, Example 2 and Example 4.

[0334] LC-MS (ESI, m / z, C 50 H 100 N2O5, 809.8, M+H)

[0335] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 4.13 (m, 2H), 3.71 - 3.30 (m, 8H), 3.10 (m, 1H), 2.54 - 2.43 (m, 3H), 2.19 - 2.13 (m, 7H), 1.60 - 1.38 (m, 22H), 1.29 - 1.26 (m, 44H), 0.93 - 0.88 (m, 12H). Pale yellow oil.

[0336] Example 32

[0337] Synthesis of Compound 29

[0338]

[0339] Chemical formula: C 57 H 112 N2O6

[0340] Molecular weight: 921.53

[0341] See the synthesis in Example 1 and Example 2.

[0342] LC-MS (ESI, m / z, C 57 H 112 N2O6, 921.8, M+H)

[0343] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 4.47 (m, 1H), 4.13 (m, 2H), 3.71 - 3.30 (m, 6H), 2.54 - 2.32 (m, 9H), 2.13 (m, 1H), 1.66 - 1.43 (m, 20H), 1.29 - 1.25 (m, 54H), 0.93 - 0.88 (m, 18H). Pale yellow oil.

[0344] Example 33

[0345] Synthesis of Compound 30

[0346]

[0347] Chemical formula: C 56 H 108 N2O6

[0348] Molecular weight: 905.49

[0349] See the synthesis of Example 1, Example 3 and Example 4.

[0350] LC-MS (ESI, m / z, C 56 H 108 N2O6, 904.8, M+H)

[0351] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 4.47 - 4.17 (m, 3H), 3.35 - 3.30 (m, 4H), 3.10 - 2.93 (m, 2H), 2.46 - 2.32 (m, 10H), 1.64 - 1.43 (m, 18H), 1.38 - 1.26 (m, 58H), 0.88 (m, 12H). Pale yellow oil.

[0352] Example 34

[0353] Synthesis of Compound 31

[0354]

[0355] Chemical formula: C 56 H 108 N2O7

[0356] Molecular weight: 921.49

[0357] See the synthesis of Example 1, Example 3 and Example 4.

[0358] LC-MS (ESI, m / z, C 56 H 108 N2O7, 921.8, M+H)

[0359] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 4.49 - 4.17 (m, 4H), 3.60 (m, 1H), 3.35 - 3.30 (m, 4H), 3.10 - 2.93 (m, 2H), 2.51 - 2.32 (m, 10H), 1.77 - 1.43 (m, 18H), 1.38 - 1.26 (m, 56H), 0.88 (m, 12H). Pale yellow oil.

[0360] Example 35

[0361] Synthesis of Compound 32

[0362]

[0363] Chemical formula: C 55 H 106 N2O7

[0364] Molecular weight: 907.46

[0365] See the synthesis of Example 1, Example 3 and Example 4.

[0366] LC-MS (ESI, m / z, C 55 H 106 N2O7, 907.8, M+H)

[0367] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.47 - 4.17 (m, 3H), 3.52 (m, 4H), 3.35 - 3.30 (m, 4H), 3.10 - 2.93 (m, 2H), 2.46 - 2.32 (m, 10H), 1.64 - 1.38 (m, 18H), 1.38 - 1.26 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0368] Example 36

[0369] Synthesis of Compound 33

[0370]

[0371] Chemical formula: C 56 H 109 N3O6

[0372] Molecular weight: 920.50

[0373] See the synthesis of Example 1, Example 3 and Example 4.

[0374] LC-MS (ESI, m / z, C 56 H 109 N3O6, 920.8, M+H)

[0375] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.47 - 4.17 (m, 3H), 3.52 (m, 4H), 3.35 - 3.30 (m, 4H), 3.10 - 2.93 (m, 2H), 2.46 - 2.29 (m, 10H), 2.14 (s, 3H), 1.64 - 1.38 (m, 18H), 1.38 - 1.26 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0376] Example 37

[0377] Synthesis of Compound 34

[0378]

[0379] Chemical formula: C 55 H 106N2O6

[0380] Molecular weight: 891.46

[0381] See the synthesis in Examples 1, 3 and 4.

[0382] LC-MS (ESI, m / z, C 55 H 106 N2O6, 891.46, M+H)

[0383] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.47 - 4.17 (m, 3H), 3.35 - 3.30 (m, 4H), 3.10 - 2.93 (m, 2H), 2.51 - 2.32 (m, 10H), 1.68 - 1.38 (m, 22H), 1.38 - 1.26 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0384] Example 38

[0385] Synthesis of Compound 35

[0386]

[0387] Chemical formula: C 55 H 108 N2O8

[0388] Molecular weight: 925.48

[0389] See the synthesis in Examples 1, 3 and 4.

[0390] LC-MS (ESI, m / z, C 55 H 108 N2O8, 925.8, M+H)

[0391] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.47 - 4.16 (m, 5H), 3.42 - 3.30 (m, 8H), 3.10 - 2.93 (m, 2H), 2.57 - 2.32 (m, 10H), 1.64 - 1.38 (m, 18H), 1.38 - 1.26 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0392] Example 39

[0393] Synthesis of Compound 36

[0394]

[0395] Chemical formula: C 53 H104 N2O6

[0396] Molecular weight: 865.42

[0397] See the synthesis in Examples 1, 3 and 4.

[0398] LC-MS (ESI, m / z, C 53 H 104 N2O6, 865.8, M+H)

[0399] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.47 - 4.16 (m, 3H), 3.35 - 3.30 (m, 4H), 3.10 - 2.93 (m, 2H), 2.43 - 2.32 (m, 6H), 2.19 (s, 6H), 1.64 - 1.38 (m, 18H), 1.38 - 1.26 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0400] Example 40

[0401] Synthesis of Compound 37

[0402]

[0403] Chemical formula: C 49 H 98 N2O4

[0404] Molecular weight: 779.33

[0405] See the synthesis in Examples 1 and 4.

[0406] LC-MS (ESI, m / z, C 49 H 98 N2O4, 779.7, M+H)

[0407] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.35 - 3.30 (m, 4H), 3.10 (m, 1H), 2.93 (m, 1H), 2.46 (m, 6H), 2.13 (m, 1H), 1.60 - 1.38 (m, 14H), 1.29 - 1.25 (m, 50H), 0.93 - 0.88 (m, 18H). Pale yellow oil.

[0408] Example 41

[0409] Synthesis of Compound 38

[0410]

[0411] Chemical formula: C 47 H 92 N2O4

[0412] Molecular weight: 749.26

[0413] See the synthesis in Examples 1 and 4.

[0414] LC-MS (ESI, m / z, C 47 H 92 N2O4, 749.7, M+H)

[0415] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.35 - 3.30 (m, 4H), 3.10 (m, 1H), 2.93 (m, 1H), 2.46 - 2.42 (m, 6H), 2.13 (m, 1H), 1.60 - 1.43 (m, 14H), 1.38 - 1.25 (m, 50H), 0.88 (m, 12H). Pale yellow oil.

[0416] Example 42

[0417] Synthesis of Compound 39

[0418]

[0419] Chemical formula: C 47 H 92 N2O5

[0420] Molecular weight: 765.26

[0421] See the synthesis in Examples 1 and 4.

[0422] LC-MS (ESI, m / z, C 47 H 92 N2O5, 765.7, M+H)

[0423] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.49 - 4.17 (m, 3H), 3.60 (m, 1H), 3.35 - 3.30 (m, 4H), 3.10 (m, 1H), 2.93 (m, 1H), 2.51 - 2.41 (m, 6H), 2.13 (m, 1H), 1.77 - 1.38 (m, 18H), 1.29 - 1.25 (m, 44H), 0.88 (m, 12H). Pale yellow oil.

[0424] Synthesis of Compound 40 in Example 43

[0425] Chemical formula: C46 H 90 N2O5

[0426]

[0427] Molecular weight: 751.24

[0428] See the synthesis in Example 1 and Example 4.

[0429] LC-MS (ESI, m / z, C 46 H 90 N2O5, 751.7, M+H)

[0430] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.52 (m, 4H), 3.35 - 3.30 (m, 4H), 3.10 (m, 1H), 2.93 (m, 1H), 2.46 - 2.40 (m, 6H), 2.13 (m, 1H), 1.60 - 1.38 (m, 14H), 1.29 - 1.25 (m, 44H), 0.88 (m, 12H). Pale yellow oil.

[0431] Example 44

[0432] Synthesis of Compound 41

[0433] Chemical formula: C 47 H 93 N3O4

[0434]

[0435] Molecular weight: 764.28

[0436] See the synthesis in Example 1 and Example 4.

[0437] LC-MS (ESI, m / z, C 47 H 93 N3O4, 764.8, M+H)

[0438] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.35 - 3.30 (m, 4H), 3.10 (m, 1H), 2.93 (m, 1H), 2.46 (m, 2H), 2.29 (s, 8H), 2.14 - 2.13 (m, 4H), 1.60 - 1.38 (m, 14H), 1.29 - 1.25 (m, 44H), 0.88 (m, 12H). Pale yellow oil.

[0439] Example 45

[0440] Synthesis of Compound 42

[0441]

[0442] Chemical formula: C 48 H 94 N2O4

[0443] Molecular weight: 763.29

[0444] See the synthesis in Example 1 and Example 4.

[0445] LC-MS (ESI, m / z, C 48 H 94 N2O4, 763.7, M+H)

[0446] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.17 (m, 2H), 3.35 - 3.30 (m, 4H), 3.10 (m, 1H), 2.93 (m, 1H), 2.51 - 2.46 (m, 6H), 2.13 (m, 1H), 1.68 - 1.38 (m, 18H), 1.29 - 1.25 (m, 48H), 0.88 (m, 12H). Pale yellow oil.

[0447] Example 46

[0448] Synthesis of Compound 43

[0449]

[0450] Chemical formula: C 46 H 92 N2O6

[0451] Molecular weight: 769.25

[0452] See the synthesis in Example 1 and Example 4.

[0453] LC-MS (ESI, m / z, C 46 H 92 N2O6, 769.8, M+H)

[0454] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.42 - 4.16 (m, 4H), 3.42 - 3.30 (m, 8H), 3.10 (m, 1H), 2.93 (m, 1H), 2.57 - 2.46 (m, 6H), 2.13 (m, 1H), 1.60 - 1.38 (m, 14H), 1.29 - 1.25 (m, 44H), 0.88 (m, 12H). Pale yellow oil.

[0455] Example 47

[0456] Synthesis of Compound 44

[0457] Chemical formula: C 55 H 112 N2O4

[0458]

[0459] Molecular weight: 865.51

[0460] Refer to the synthesis in Example 1 and Example 4.

[0461] LC-MS (ESI, m / z, C 55 H 112 N2O4, 865.8, M+H)

[0462] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 3.71 - 3.46 (m, 2H), 3.35 - 3.30 (m, 8H), 3.10 (m, 2H), 2.54 - 2.46 (m, 7H), 1.50 - 1.38 (m, 22H), 1.29 - 1.25 (m, 52H), 0.93 - 0.88 (m, 18H). Pale yellow oil.

[0463] Example 48

[0464] Synthesis of Compound 45

[0465]

[0466] Chemical formula: C 56 H 112 N2O4

[0467] Molecular weight: 877.52

[0468] Refer to the synthesis in Example 1 and Example 4.

[0469] LC-MS (ESI, m / z, C 56 H 112 N2O4, 877.9, M+H)

[0470] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 3.71 - 3.46 (m, 2H), 3.35 - 3.30 (m, 8H), 3.10 (m, 2H), 2.54 - 2.42 (m, 7H), 1.50 - 1.37 (m, 26H), 1.29 - 1.25 (m, 54H), 0.88 (m, 12H). Pale yellow oil.

[0471] Example 49

[0472] Synthesis of Compound 46

[0473]

[0474] Chemical formula: C 56 H 112 N2O5

[0475] Molecular weight: 893.52

[0476] See the synthesis in Example 1 and Example 4.

[0477] LC-MS (ESI, m / z, C 56 H 112 N2O5, 890.9, M+H)

[0478] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 4.49 (m, 1H), 3.71 - 3.46 (m, 3H), 3.35 - 3.30 (m, 8H), 3.10 (m, 2H), 2.54 - 2.41 (m, 7H), 1.77 - 1.38 (m, 26H), 1.29 - 1.25 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0479] Example 50

[0480] Synthesis of Compound 47

[0481]

[0482] Chemical formula: C 55 H 110 N2O5

[0483] Molecular weight: 879.49

[0484] See the synthesis in Example 1 and Example 4.

[0485] LC-MS (ESI, m / z, C 55 H 110 N2O5, 879.8, M+H)

[0486] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 3.71 - 3.46 (m, 6H), 3.35 - 3.30 (m, 8H), 3.10 (m, 2H), 2.54 - 2.40 (m, 7H), 1.50 - 1.38 (m, 22H), 1.29 - 1.25 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0487] Example 51

[0488] Synthesis of Compound 48

[0489]

[0490] Chemical formula: C 56 H 113 N3O4

[0491] Molecular weight: 892.54

[0492] See the synthesis in Example 1 and Example 4.

[0493] LC-MS (ESI, m / z, C 56 H 113 N3O4, 892.9, M+H)

[0494] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 3.71 - 3.30 (m, 10H), 3.10 (m, 2H), 2.54 - 2.46 (m 3H), 2.29 (s, 8H), 2.14 (s, 3H), 1.50 - 1.38 (m, 22H), 1.29 - 1.25 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0495] Example 52

[0496] Synthesis of Compound 49

[0497]

[0498] Chemical formula: C 55 H 110 N2O4

[0499] Molecular weight: 863.50

[0500] See the synthesis in Example 1 and Example 4.

[0501] LC-MS (ESI, m / z, C 55 H 110 N2O4, 863.9, M+H)

[0502] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 3.71 - 3.30 (m, 10H), 3.10 (m, 2H), 2.51 - 2.46 (m, 7H), 1.68 (m, 4H), 1.50 - 1.38 (m, 22H), 1.29 - 1.25 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0503] Example 53

[0504] Synthesis of Compound 50

[0505]

[0506] Chemical formula: C 55 H 112 N2O6

[0507] Molecular weight: 897.51

[0508] Refer to the synthesis in Example 1 and Example 4.

[0509] LC-MS (ESI, m / z, C 55 H 112 N2O6, 897.9, M+H)

[0510] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 4.16 (m, 2H), 3.71 - 3.35 (m, 14H), 3.10 (m, 2H), 2.57 - 2.46 (m, 7H), 1.50 - 1.38 (m, 22H), 1.29 - 1.25 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0511] Example 54

[0512] Synthesis of Compound 51

[0513]

[0514] Chemical formula: C 53 H 108 N2O4

[0515] Molecular weight: 837.46

[0516] Refer to the synthesis in Example 1 and Example 4.

[0517] LC-MS (ESI, m / z, C 53 H 108 N2O4, 837.9, M+H)

[0518] HNMR (DMSO-d6, 400 MHz) δ8.01 (s, 1H), 3.71 - 3.30 (m, 10H), 3.10 (m, 2H), 2.54 - 2.43 (m, 3H), 2.19 (s, 6H), 1.50 - 1.38 (m, 22H), 1.29 - 1.25 (m, 52H), 0.88 (m, 12H). Pale yellow oil.

[0519] Example 55

[0520] Synthesis of Compound 52

[0521]

[0522] Chemical formula: C 50 H 102 N2O3

[0523] Molecular weight: 779.38

[0524] See the synthesis in Example 1 and Example 4.

[0525] LC-MS (ESI, m / z, C 50 H 102 N2O3, 779.8, M+H)

[0526] 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (s, 1H), 3.71 - 3.30 (m, 8H), 3.10 (m, 1H), 2.54 - 2.46 (m, 7H), 1.50 - 1.38 (m, 14H), 1.29 - 1.26 (m, 56H), 0.93 - 0.88 (m, 15H). Pale yellow oil.

[0527] Example 56

[0528] Synthesis of Compound 53

[0529]

[0530] Chemical formula: C 48 H 95 NO4

[0531] Molecular weight: 750.29

[0532] See the synthesis in Example 1 and Example 4.

[0533] LC-MS (ESI, m / z, C 48 H 95 NO4, 750.7, M+H)

[0534] 1H NMR (DMSO-d6, 400 MHz) δ 4.18 (m, 2H), 3.83 - 3.58 (m, 2H)03.35 (m, 4H), 3.10 - 2.97 (m, 3H), 2.43 - 2.42 (m, 5H), 1.60 - 1.37 (m, 20H), 1.37 - 1.25 (m, 50H), 0.88 (m, 9H). Pale yellow oil.

[0535] Example 57

[0536] Synthesis of Compound 54

[0537]

[0538] Chemical formula: C 47 H 93 NO5

[0539] Molecular weight: 752.26

[0540] See the synthesis in Examples 1 and 4.

[0541] LC-MS (ESI, m / z, C 47 H 93 NO5, 752.7, M+H)

[0542] 1H NMR (DMSO-d6, 400 MHz) δ 4.18 (m, 2H), 3.83 - 3.57 (m, 6H), 3.35 (m, 4H), 3.10 - 2.97 (m, 3H), 2.50 - 2.43 (m, 5H), 1.60 - 1.38 (m, 14H), 1.29 - 1.25 (m, 50H), 0.88 (m, 9H). Pale yellow oil.

[0543] Example 58

[0544] Synthesis of Compound 55

[0545]

[0546] Chemical formula: C 48 H 96 N2O4

[0547] Molecular weight: 765.31

[0548] See the synthesis in Examples 1 and 4.

[0549] LC-MS (ESI, m / z, C 48 H 96 N2O4, 765.7, M+H)

[0550] 1H NMR (DMSO-d6, 400 MHz) δ 4.18 (m, 2H), 3.83 - 3.58 (m, 2H), 3.35 (m, 4H), 3.10 - 2.97 (m, 3H), 2.43 - 2.29 (m, 9H), 2.14 (s, 3H), 1.60 - 1.38 (m, 14H), 1.29 - 1.25 (m, 50H), 0.88 (m, 9H). Pale yellow oil.

[0551] Example 59

[0552] Synthesis of Compound 56

[0553]

[0554] Chemical formula: C 47 H 95 NO6

[0555] Molecular weight: 770.28

[0556] See the synthesis in Example 1 and Example 4.

[0557] LC-MS (ESI, m / z, C 47 H 95 NO6, 770.7, M+H)

[0558] 1H NMR (DMSO-d6, 400 MHz) δ 4.18 - 4.16 (m, 4H), 3.83 - 3.35 (m, 10H), 3.10 - 2.97 (m, 3H), 2.57 - 2.43 (m, 5H), 1.60 - 1.38 (m, 14H), 1.29 - 1.25 (m, 50H), 0.88 (m, 9H). Pale yellow oil.

[0559] Example 60

[0560] In vivo evaluation of luciferase mRNA using lipid nanoparticle compositions

[0561] Dissolve cationic lipid, DSPC, cholesterol, and PEG-lipid in ethanol at a molar ratio of 50:10:38:2 or 48:10:40:2. Prepare lipid nanoparticles (LNPs) at a total lipid to mRNA weight ratio of approximately 10:1 to 30:1. Briefly, dilute the mRNA to 0.15 mg / mL in 10 mL to 50 mL citrate buffer (pH = 4). Using a syringe pump, mix the ethanol solution of the lipid with the aqueous solution of the mRNA at a ratio of approximately 1:5 to 1:3 (volume / volume), with a total flow rate of more than 10 mL / min. Then remove the ethanol and replace the external buffer with PBS by dialysis. Finally, filter the lipid nanoparticles through a sterile filter with a pore size of 0.2 μm. The particle size of the lipid nanoparticles measured by quasi-elastic light scattering using a Malvern Zetasizer Nano ZS is approximately 65 - 105 nm in diameter, and in some cases, approximately 75 - 100 nm in diameter.

[0562] Studies were conducted on female C57BL / 6 mice at 6 - 8 weeks of age and CD-1 mice at 8 - 10 weeks of age according to the guidelines established by the National Science and Technology Commission. Different doses of mRNA lipid nanoparticles were administered systemically via tail vein injection, and the animals were euthanized at specific time points after dosing (e.g., 5 hours). The liver and spleen were collected in pre-weighed tubes, the weights were determined, and they were immediately snap-frozen in liquid nitrogen and stored at -80 °C until used for analysis.

[0563] For the liver, approximately 50 mg was cut for analysis in a 2 mL FastPrep tube (MP Biomedicals, Solon OH). A 1 / 4" ceramic ball (MP Biomedicals) was added to each tube, and 500 μL of Glo Lysis Buffer - GLB (Promega, Madison WI), equilibrated to room temperature, was added to the liver tissue. The liver tissue was homogenized using a FastPrep24 instrument (MP Biomedicals) at 2 × 6.0 m / s for 15 seconds. The homogenate was incubated at room temperature for 5 minutes, then diluted 1:4 in GLB and evaluated using the SteadyGlo Luciferase Assay System (Promega). Specifically, 50 μL of the diluted tissue homogenate was reacted with 50 μL of the SteadyGlo substrate, shaken for 10 seconds, then incubated for 5 minutes, and then quantified using a SpectraMAX_L chemiluminescent microplate reader (Molecular Devices (Shanghai) Co., Ltd.). The amount of protein in the assay was determined using a BCA Protein Quantification Kit (Shanghai Yise Medical Technology Co., Ltd.). The relative light units (RLU) were then normalized to the total μg of protein assayed. To convert RLU to μg of luciferase, a standard curve was generated using QuantiLum Recombinant Luciferase (Promega).

[0564] The FLuc mRNA (L-6107) from Trilink Biotechnologies will express the luciferase protein, which was originally isolated from the firefly (Photinus pyralis). Fluc is commonly used in mammalian cell cultures to measure gene expression and cell viability. It emits bioluminescence in the presence of the substrate luciferin. This capped and polyadenylated mRNA is completely substituted with 5-methylcytidine and pseudouridine.

[0565] Example 61

[0566] Determination of the pKa of the formulated lipids

[0567] The pKa of the formulated cationic lipid is related to the efficacy of the LNP used for nucleic acid delivery. The preferred pKa range is 5 - 7. The pKa of each cationic lipid is determined in lipid nanoparticles using an assay based on the fluorescence of 2-(p-toluidino)-6-naphthalenesulfonic acid (TNS). As described in Example 60, an ordered method is used to prepare lipid nanoparticles containing cationic lipid / DSPC / cholesterol / PEG lipid (50 / 10 / 38 / 2 mol%) at a total lipid concentration of 0.4 mM in PBS. TNS is prepared as a 100 μM stock solution in distilled water. The vesicles are diluted to contain 24 μM lipid in 2 mL of buffer solution containing 10 mM HEPES, 10 mM MES, 10 mM ammonium acetate, and 130 mM NaCl, with a pH value ranging from 2.5 to 11. Equal aliquots of the TNS solution are added to give a final concentration of 1 μM, and after vortex mixing, the fluorescence intensity is measured at room temperature in an SLM Aminco Series 2 luminescence spectrophotometer using excitation and emission wavelengths of 321 nm and 445 nm. A sigmoidal best-fit analysis is applied to the fluorescence data, and the pKa is measured as the pH that produces half-maximal fluorescence intensity.

[0568] Example 62

[0569] The efficacy of lipid nanoparticle formulations containing various cationic lipids was determined using a rodent model of in vivo luciferase mRNA expression.

[0570] For comparison purposes, as described in Example 60, these lipids were also used to formulate lipid nanoparticles containing FLuc mRNA (L-6107) using an ordered mixing method. Lipid nanoparticles were formulated using the following molar ratios: 50% cationic lipid / 10% distearoylphosphatidylcholine (DSPC) / 38% cholesterol / 2% PEG lipid ("PEG-DMG", i.e., (1-(monomethoxy-polyethylene glycol)-2,3-dimyristoyl glycerol, with an average PEG molecular weight of 2000). As described in Example 60, the relative activity was determined by measuring luciferase expression in the liver 5 hours after administration via tail vein injection. The activity was compared at doses of 0.3 and 1.0 mg mRNA / kg and expressed as ng luciferase / g liver measured 5 hours after administration as described in Example 60. The results of Examples 61 and 62 are shown in Table 2.

[0571] Table 2 Comparison of lipids showing activity with mRNA

[0572]

[0573]

[0574]

[0575]

[0576]

[0577]

[0578]

[0579]

[0580] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as the scope described in this specification.

[0581] The above-described embodiments merely represent several implementation manners of the present invention. The description thereof is relatively specific and detailed, but it should not be construed as a limitation on the scope of the disclosed patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the present invention patent shall be subject to the appended claims.

Claims

1. A cationic lipid compound, which is a compound having one of the following structures: Compound 27 ; Compound 31: ; Compound 32 ; Compound 33 ; Compound 34 ; Compound 35 ; Compound 38: ; Compound 39: ; Compound 43 ; Compound 44 ; Compound 46 ; Compound 47 ; Compound 48 ; Compound 49 ; Compound 50 ; Compound 51: .

2. A composition comprising the compound according to claim 1 and a therapeutic agent and / or a prophylactic agent.

3. The composition according to claim 2, which further comprises one or more components selected from neutral lipids, steroids, and polymeric lipids.

4. The composition according to claim 3, wherein the neutral lipid is one or more components selected from DSPC, DPPC, DMPC, DOPC, POPC, DOPE, and SM.

5. The composition according to claim 4, wherein the neutral lipid is DSPC.

6. The composition according to any one of claims 3-5, wherein the molar ratio of the compound to the neutral lipid is from 2:1 to 8:

1.

7. The composition according to claim 3, wherein the steroid is one or more of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, and brassicasterol.

8. The composition according to claim 7, wherein the steroid is cholesterol.

9. The composition according to any one of claims 7-8, wherein the molar ratio of the compound to the steroid is from 1:1 to 5:

1.

10. The composition according to claim 3, wherein the polymeric lipid is a polyethylene glycolated lipid.

11. The composition according to claim 10, wherein the polyethylene glycolated lipid is PEG-DAG, PEG-PE, PEG-S-DAG, PEG-cer, or PEG-dialkoxypropyl carbamate.

12. The composition according to claim 2, wherein the therapeutic agent is a nucleic acid.

13. The composition according to claim 12, wherein the nucleic acid is selected from antisense nucleic acid, small interfering nucleic acid (siRNA), microRNA (miRNA), and messenger nucleic acid (mRNA).

14. Use of the composition according to any one of claims 2-13 in the preparation of a medicament or a vaccine for treating a disease selected from cancer, proliferative diseases, neurodegenerative diseases, or metabolic diseases.

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