Pyrrolidine lipid compounds, preparation methods thereof, compositions and applications

Abstract

The present invention discloses a novel pyrrolidine compound represented by Formula A, or its isomer, or its N-oxide, or its pharmaceutically acceptable salt, prodrug. Also disclosed are a preparation method, a composition and an application of the above pyrrolidine compound. As a cationic lipid, this kind of compound is suitable for a pharmaceutical composition for nucleic acid delivery, and can achieve better stability, pKa value and intracellular delivery efficiency of the drug, 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 compositions of the present invention may also 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 presents 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-containing nanoparticle compositions, liposomes, and lipoplexes can be used effectively 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 including polyunsaturated lipids (such as phospholipids), structural lipids (such as steroids), and / or lipids containing polyethylene glycol (lipid-polymer conjugates). Cationic lipids include amine-containing lipids that can be easily protonated.

[0004] However, the use of oligonucleotides in a therapeutic setting currently faces two problems. First, free RNA is susceptible to nuclease digestion in plasma. Second, the ability of free RNA to enter intracellular compartments where the relevant translation machinery exists 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 impose unacceptable toxicity and / or risk on the patient. The present invention provides these and related advantages. Summary of the Invention

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

[0007] A pyrrolidine compound represented by formula A, or an isomer thereof, or an N-oxide thereof, or a pharmaceutically acceptable salt or prodrug thereof;

[0008]

[0009] Wherein:

[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-, L2 is selected from the following structures: -C-, -(C=O)O-, -C(=O)-, -S(O) X -, -C(=O)S-, -C(=O)-N-;

[0011] R1 and R2 are each independently a C6-C24 alkyl or C6-C24 alkenyl group, 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 group, 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, and 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 group;

[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 List of Compounds

[0018]

[0019]

[0020]

[0021]

[0022]

[0023]

[0024]

[0025]

[0026]

[0027]

[0028]

[0029]

[0030]

[0031]

[0032] In some embodiments, there are provided compositions comprising any one or more of the compounds of structural formula (I) and a therapeutic and / or prophylactic agent.

[0033] In some embodiments, there are provided compositions comprising any one or more of the compounds of structure (I) and a therapeutic and / or prophylactic agent. In some embodiments, the composition comprises any one of the compounds of structure (I) and a therapeutic and / or 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 the various embodiments of the composition.

[0034] In some embodiments, the neutral lipid is 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).

[0035] In some embodiments, the steroid is selected from cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid, α-tocopherol, and mixtures thereof. In some embodiments, the preferred steroid is cholesterol.

[0036] In some embodiments, the pegylated lipid is 1,2-dimyristoyl-sn-glycero-3-methoxypolyethylene glycol (PEG-DMG).

[0037] In some embodiments, the composition ratios are in the following ranges: about 10 to 60 mol% of the compound, about 0 to 30 mol% of neutral lipid, about 10 to 55 mol% of steroid, and about 0 to 10 mol% of polymer-conjugated lipid.

[0038] 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.

[0039] 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.

[0040] 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 concentrations and doses.

[0041] The administration of the compositions of the present invention can be carried out by any acceptable mode of administration for a reagent 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, inhaled, 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 a subject. The composition to be administered to an object 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 a compound in aerosol form according to 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 a relevant disease or condition according to the teachings of the present invention.

[0042] 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 inhaled administration.

[0043] When intended for oral administration, the pharmaceutical composition is preferably in solid or liquid form, where forms considered to be solid or liquid herein include semi-solid, semi-liquid, suspension and gel forms.

[0044] As a solid composition for oral administration, the pharmaceutical composition can be formulated into forms such as 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.

[0045] When the pharmaceutical composition is in the form of a capsule, it can contain a liquid carrier in addition to the materials of the above types, such as polyethylene glycol or oil.

[0046] The pharmaceutical composition can be in liquid form, such as syrup, solution, emulsion or suspension. As two examples, the liquid can be used 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 a sweetening agent, a preservative, a coloring agent, and a flavor enhancer. In the composition for administration by injection, one or more of a surfactant, a preservative, a wetting agent, a dispersing agent, a suspending agent, a buffering agent, a stabilizing agent, and an isotonic agent can be included.

[0047] The liquid pharmaceutical composition of the present invention, whether it is a solution, a suspension or other similar form, can include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, 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. The parenteral preparation can be encapsulated in ampoules, disposable syringes or multi-dose bottles made of glass or plastic. Physiological saline is the preferred adjuvant. The injectable pharmaceutical composition is preferably sterile.

[0048] The pharmaceutical composition of the present invention can 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 compound of the present invention can be delivered in a single-phase, two-phase or three-phase system in order to deliver the active ingredient. The delivery of the aerosol includes the necessary container, activator, valve, sub-container, etc., which together can form a kit. A person skilled in the art can determine the preferred aerosol without undue experimentation.

[0049] The pharmaceutical composition of the present invention can be prepared by methods well known in the pharmaceutical field. The pharmaceutical composition intended for administration 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. A surfactant can be added to facilitate the formation of a uniform solution or suspension. A surfactant is a compound that interacts non-covalently with the compound of the present invention in order to facilitate the dissolution or uniform suspension of the compound in an aqueous delivery system.

[0050] 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 rate of excretion; the drug combination; the severity of the particular case, etc.

[0051] 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 agent can be administered in a different oral dosage formulation. When different dosage formulations are used, 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 therapies include all of these dosing regimens.

[0052] 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 nanoliposome compositions, enabling 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.

[0053] The preparation methods of the above-mentioned compounds and compositions are described below and / or are known in the art.

[0054] 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 aryl ester. The protecting groups can be added or removed according to standard techniques, which are known to those skilled in the art and described herein.

[0055] 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". Therefore, prodrugs of the compounds of the present invention are included within the scope of the present invention.

[0056] In addition, all compounds of the present invention in the form of free base or free acid 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.

[0057] The following examples are provided by way of illustration and not limitation.

[0058] In the following examples, unless otherwise indicated, all solvents and reagents used are commercially available and used as received.

[0059] The procedures described below can be used to synthesize Compound 1.

[0060] The following abbreviations are used herein:

[0061] DIEA: Diisopropylethylamine

[0062] HATU: 2-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0063] K2CO3: Potassium carbonate

[0064] DMF: N,N-Dimethylformamide

[0065] NaOH: Sodium hydroxide

[0066] THF: Tetrahydrofuran

[0067] TBAB: Tetrabutylammonium bromide Detailed Description of the Invention

[0068] Example 1:

[0069] Representative Route

[0070] Compound CLPP: N-(Diethylamino)ethyl)-1-pentadecanoyl-4-pentadecyloxypyrrolidine-2-carboxamide

[0071] 1) Preparation of Intermediate F

[0072]

[0073] At room temperature, raw material H (1 g, 4.32 mmol), G (1.51 g, 5.18 mmol) and DMF (100 ml) were added to a 250 ml four-necked flask and stirred until dissolved. K2CO3 (0.78 g, 5.62 mmol) was added, and the mixture was heated to 80 °C and stirred for 4 h. The reaction was monitored by TLC until completion. Ethyl acetate was added, and the organic phase was washed successively with water, sodium bicarbonate, dilute hydrochloric acid, and water. After drying over Na2SO4, the mixture was concentrated under reduced pressure to obtain a crude product of intermediate F. The crude product was purified by column chromatography to obtain 1.49 g of the target compound with a yield of 78%.

[0074] Molecular formula: C 25 H 47 NO5

[0075] Molecular weight: 441.64

[0076] LC-MS: m / z 442 [M+H]

[0077] 1H NMR (DMSO-d6): δ (ppm) 11.02 (s, 1H) 4.25 (t, 1H, J = 3.5), 3.50 (m, 2H) 3.39 (m, 2H), 3.05 (m, 1H), 1.98 (m, 2H), 1.40 - 1.47 (m, 11H), 1.25 - 1.33 (m, 24H), 0.99 (m, 3H).

[0078] 2) Preparation of intermediate D

[0079]

[0080] (DMF) 20 mL, cooled in an ice bath, stirred, and ≤5 °C, 0.84 g (6.5 mmol) of diisopropylethylamine (DIEA), intermediate N-substituted amine (E, 2.34 mmol) and 0.83 g (2.6 mmol) of O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) were added successively. Stir at room temperature for 3 h, and monitor the reaction by TLC until completion. Under stirring, the reaction solution was poured into 100 g of ice water, and stirring was continued for 0.5 h. Extracted with ethyl acetate 3 times (30 mL × 3), the ethyl acetate layers were combined, and the organic phase was washed successively with water, hydrochloric acid, sodium bicarbonate, and brine, dried over anhydrous MgSO4, and distilled under reduced pressure to obtain a crude product of intermediate C. Column chromatography (eluted with petroleum ether:ethyl acetate = 5:1) gave 1.07 g of a light-colored oil with a yield of 85.01%.

[0081] Molecular formula: C 31 H 61 N3O4

[0082] Molecular weight: 539.47

[0083] LC-MS: m / z 540 [M+H]

[0084] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.27 (t, 1H, J=3.5 Hz), 3.52 (m, 2H) 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J=3 Hz), 2.70 (m, 2H), 2.30 (m, 4H), 1.98 (m, 2H), 1.40 - 1.47 (m, 11H), 1.25 - 1.33 (m, 24H), 1.02 - 0.99 (m, 9H).

[0085] 3) Preparation of Intermediate C

[0086]

[0087] Place (3.13 g, 5.8 mmol) of Intermediate C and 30 mL of ethyl acetate in a 100 mL three-necked flask, stir, cool in an ice-water bath to ≤10 °C, add 2.5 mL of 33% HCl ethanol solution (14.5 mmol) dropwise within 0.5 h, stir at room temperature for 4 h, detect the completion of the reaction by TLC, extract with water 3 times (30 mL × 3), combine the aqueous layers, wash the aqueous phase with ethyl acetate, add 30 mL of dichloromethane to the aqueous layer, stir, adjust the pH to 8 - 9 with Na2CO3, separate the layers, extract the aqueous layer with dichloromethane 2 times (30 ml × 2), combine the dichloromethane layers, wash with saturated brine 3 times, dry over anhydrous MgSO4, filter, concentrate under reduced pressure to obtain the crude product of Intermediate C, and purify by column chromatography to obtain 2.45 g of an oily substance with a yield of 96%.

[0088] Molecular formula: C 26 H 53 N3O2

[0089] Molecular weight: 439.72

[0090] LC-MS: m / z 440 [M+H]

[0091] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.27 (t, 1H, J=3.5), 3.52 (m, 2H) 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J=3), 2.70 (m, 2H), 2.30 (m, 4H), 2.01 (s, 1H), 1.98 (m, 2H), 1.47 (m, 2H), 1.25 - 1.33 (m, 24H), 0.99 (m, 9H).

[0092] 4) Preparation of Compound 1

[0093]

[0094] Into a 100 mL reaction flask, add intermediate C (1.03 g, 2.34 mmol) and 20 mL of N,N-dimethylformamide (DMF). Cool in an ice bath, stir, and sequentially add 0.84 g (6.5 mmol) of diisopropylethylamine (DIEA), intermediate CLPP-6 (2.34 mmol), and 0.83 g (2.6 mmol) of O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) at ≤5°C. Stir at room temperature for 3 h, and detect by TLC that the reaction is complete. Under stirring, pour the reaction solution into 100 g of ice water, continue to stir for 0.5 h, extract with ethyl acetate 3 times (30 mL × 3), combine the ethyl acetate layers, wash the organic phase successively with water, hydrochloric acid, sodium bicarbonate, and brine, dry over anhydrous MgSO4, and after distillation under reduced pressure, obtain the crude product of compound 1. Purify by column chromatography (eluting with petroleum ether:ethyl acetate = 5:1) to obtain 1.4 g of a pale yellow oil, with a yield of 90%.

[0095] Molecular formula: C 41 H 81 N3O3

[0096] Molecular weight: 664.10

[0097] LC-MS: m / z 665 [M+H]

[0098] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.27 (t, 1H, J = 3.5), 3.52 (m, 2H) 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J = 3), 2.70 (m, 2H), 2.36 (t, 2H, J = 6), 2.30 (m, 4H), 2.20 (m, 2H), 1.60 (m, 2H), 1.47 (m, 2H), 1.25 - 1.33 (m, 42H), 0.99 (m, 12H).

[0099] Synthesis of 6-bromohexyl 2-hexyldecanoate (C1) in Example 2

[0100]

[0101] Into a 500 mL reaction flask, commercially available 2 - hexyldecanoic acid (10 g, 35.15 mmol) and 200 mL of tetrahydrofuran (THF) were added. Subsequently, 8.27 g (45.70 mmol) of 6 - bromohexanol and 6.3 g of potassium carbonate were added successively. The temperature was raised to 68 °C and stirred for 8 h. The reaction was monitored by TLC and found to be complete. Under stirring, the mixture was extracted with ethyl acetate three times (30 mL × 3). The ethyl acetate layers were combined, and the organic phase was washed successively with water, hydrochloric acid, sodium bicarbonate, and brine, dried over anhydrous MgSO4, and then distilled under reduced pressure to obtain a crude product of 6 - bromohexyl 2 - octyldecanoate. After column chromatography (eluted with petroleum ether:ethyl acetate = 10:1), 12.56 g of 6 - bromohexyl 2 - hexyldecanoate as a yellow oil was obtained, with a yield of 80%.

[0102] Molecular formula: C 22 H 43 BrO2

[0103] Molecular weight: 419.54

[0104] LC - MS: m / z 420 [M + H]

[0105] 1H NMR (DMSO - d6): 4.13 (m, 2H,), 3.52 (m, 2H) 2.13 (m, 1H), 1.82 (m, 2H), 1.60 (m, 6H), 1.47 (m, 2H), 1.29 (m, 16H), 0.90 (m, 6H).

[0106] Synthesis of Compound 6 - (heptadec - 9 - yloxy) - 6 - oxohexanoic acid (C2) in Example 3

[0107]

[0108] Into a 500 mL reaction flask, commercially available 9 - heptadecanol (10 g, 38.99 mmol) and 100 mL of N,N - dimethylformamide (DMF) were added. Subsequently, 5.7 g (38.99 mmol) of adipic acid and sodium hydroxide (2.34 g, 58.49 mmol) were added successively. The temperature was raised to 80 °C and stirred for 6 h. The reaction was monitored by TLC and found to be complete. Under stirring, the mixture was extracted with ethyl acetate three times (30 mL × 3). The ethyl acetate layers were combined, and the organic phase was washed successively with water, sodium bicarbonate, hydrochloric acid, and brine, dried over anhydrous MgSO4, and then distilled under reduced pressure to obtain a crude product of 6 - (heptadec - 9 - yloxy) - 6 - oxohexanoic acid. After column chromatography (eluted with petroleum ether:ethyl acetate = 1:1), 9 g of 6 - (heptadec - 9 - yloxy) - 6 - oxohexanoic acid as a yellow oil was obtained, with a yield of 60.03%.

[0109] Molecular formula: C 23 H 44 O4

[0110] Molecular weight: 384.60

[0111] LC-MS: m / z 385 [M+H]

[0112] 1H NMR (DMSO-d6): 11.87 (s, 1H), 4.47 (m, 1H), 2.32 (m, 2H), 2.21 (m, 2H), 1.64 (m, 2H), 1.52 (m, 2H), 1.49 (m, 4H), 1.29 (m, 4H), 1.26 (m, 20H), 0.90 (m, 6H).

[0113] Synthesis of Compound 6-Bromohexyl 9-Heptadecyl Ether (C3) in Example 4

[0114]

[0115] Into a 500 ml reaction flask, commercially available 9-heptadecanol (10 g, 38.99 mmol) and 50 mL of N,N-dimethylformamide (DMF) were added. Subsequently, 5.7 g (38.99 mmol) of 6-bromohexanol, sodium hydroxide (2.34 g, 58.49 mmol) and 2.34 g of water were added in sequence. 0.3 equivalent of TBAB was added, and the temperature was raised to 60 °C and stirred for 12 h. The reaction was monitored by TLC and found to be complete. Under stirring, the mixture was extracted with ethyl acetate three times (30 mL × 3). The ethyl acetate layers were combined, and the organic phase was washed successively with water, sodium bicarbonate, hydrochloric acid, and brine, and dried over anhydrous MgSO4. After distillation under reduced pressure, the crude product of the side chain 6-bromohexyl 9-heptadecyl ether was obtained. Column chromatography (eluted with petroleum ether:ethyl acetate = 20:1) gave 11.4 g of a yellow oil of 6-(heptadecyl-9-oxy)-6-oxohexanoic acid, with a yield of 70%.

[0116] Molecular formula: C 23 H 47 BrO

[0117] Molecular weight: 419.53

[0118] LC-MS: m / z 420 [M+H]

[0119] 1H NMR (DMSO-d6): 3.52 (m, 2H), 3.35 (m, 2H), 3.10 (m, 1H), 1.82 (m, 2H), 1.52 (m, 2H), 1.49 (m, 2H), 1.38 (m, 4H), 1.29 (m, 6H), 1.26 (m, 20H), 0.90 (m, 6H).

[0120] Compounds of Examples 2 to 60 were prepared by a synthetic method similar to that of Examples 1, 2, 3, and 4.

[0121] Synthesis of Compound 2 in Example 5

[0122]

[0123] A light yellow compound 2 was prepared by a synthesis method similar to that in Example 1

[0124] Molecular formula: C 42 H 83 N3O3

[0125] Molecular weight: 678.13

[0126] LC-MS: m / z 679 [M+H]

[0127] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.27 (t, 1H, J = 3.5 Hz), 3.52 (m, 2H), 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J = 3 Hz), 2.70 (m, 2H), 2.36 (m, 6H), 2.30 (m, 4H), 2.20 (m, 2H), 1.60 (m, 2H), 1.47 (m, 2H), 1.25 - 1.33 (m, 46H), 0.99 (m, 12H).

[0128] Synthesis of Compound 3 in Example 6

[0129]

[0130] A light yellow compound 3 was prepared by a synthesis method similar to that in Example 1

[0131] Molecular formula: C 44 H 87 N3O3

[0132] Molecular weight: 706.18

[0133] LC-MS: m / z 707 [M+H]

[0134] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.27 (t, 1H, J = 3.5), 3.52 (m, 2H), 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J = 3), 2.70 (m, 2H), 2.36 (m, 6H), 2.30 (m, 6H), 2.20 (m, 2H), 1.60 (m, 2H), 1.47 (m, 2H), 1.25 - 1.33 (m, 50H), 0.99 (m, 12H).

[0135] Synthesis of Compound 4 in Example 7

[0136]

[0137] The light yellow compound 4 was prepared by a synthetic method similar to that in Example 1

[0138] Molecular formula: C 44 H 83 N3O3

[0139] Molecular weight: 702.15

[0140] LC-MS: m / z 703 [M+H]

[0141] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 5.60 (d, 2H, J = 2.6 Hz), 5.44 (m, 2H), 54.27 (t, 1H, J = 3.5), 3.52 (m, 2H), 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J = 3), 2.70 (m, 2H), 2.65 (m, 2H), 2.36 (t, 2H, J = 6), 2.30 (m, 4H), 2.20 (m, 2H), 1.60 (m, 2H), 1.47 (m, 2H), 1.25 - 1.33 (m, 36H), 0.99 (m, 12H).

[0142] Synthesis of Compound 5 in Example 8

[0143]

[0144] The light yellow compound 5 was prepared by a synthetic method similar to that in Example 1

[0145] Molecular formula: C 44 H 89 N3O2

[0146] Molecular weight: 691.70

[0147] LC-MS: m / z 692 [M+H]

[0148] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 3.37 (m, 2H), 3.30 (m, 2H) 3.23 (m, 1H), 2.63 (m, 2H), 2.44 (m, 4H), 2.36 (m, 4H), 1.47 (m, 2H), 1.40 (m, 2H), 1.25 - 1.33 (m, 48H), 0.99 (m, 12H).

[0149] Synthesis of Compound 6 in Example 9

[0150]

[0151] A light yellow compound 6 was prepared by a synthetic method similar to that in Example 1

[0152] Molecular formula: C 41 H 83 N3O2

[0153] Molecular weight: 650.12

[0154] LC-MS: m / z 651 [M+H]

[0155] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 3.37 (m, 2H), 3.30 (m, 2H) 3.23 (m, 1H), 2.63 (m, 2H), 2.44 (m, 4H), 2.36 (m, 4H), 1.47 (m, 2H), 1.40 (m, 2H), 1.25 - 1.33 (m, 42H), 0.99 (m, 12H).

[0156] Synthesis of Compound 7 in Example 10

[0157]

[0158] A light yellow compound 7 was prepared by a synthetic method similar to that in Example 1

[0159] Molecular formula: C 44 H 87 N3O3

[0160] Molecular weight: 706.18

[0161] LC-MS: m / z 707 [M+H]

[0162] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.46 (t, 1H, J = 5), 3.62 (m, 2H), 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J = 3), 2.70 (m, 2H), 2.45 (m, 1H), 2.36 (t, 2H), 2.30 (m, 4H), 2.20 (m, 2H), 1.5 - 1.25 (m, 54H), 0.99 (m, 12H).

[0163] Synthesis of Compound 8 in Example 11

[0164]

[0165] A light yellow compound 8 was prepared by a synthetic method similar to that in Example 1

[0166] Molecular formula: C32 H 63 N3O3

[0167] Molecular weight: 537.93

[0168] LC-MS: m / z 540 [M+H]

[0169] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.46 (t, 1H, J = 5), 3.62 (m, 2H), 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J = 3), 2.70 (m, 2H), 2.45 (m, 1H), 2.36 (t, 2H), 2.30 (m, 4H), 2.20 (m, 2H), 1.5 - 1.25 (m, 32H), 0.99 (m, 9H).

[0170] Synthesis of Compound 9 in Example 12

[0171]

[0172] A light yellow compound 9 was prepared by a synthetic method similar to that in Example 1 and Example 2

[0173] Molecular formula: C 48 H 95 N3O4

[0174] Molecular weight: 778.34

[0175] LC-MS: m / z 779 [M+H]

[0176] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.46 (t, 1H, J = 5), 3.80 (m, 2H), 3.62 (m, 2H), 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J = 3), 2.70 (m, 2H), 2.45 (m, 1H), 2.36 (t, 2H, J = 6), 2.30 (m, 4H), 2.20 (m, 2H), 1.5 - 1.25 (m, 50H), 0.90 (m, 15H).

[0177] Synthesis of Compound 10 in Example 13

[0178]

[0179] A yellow compound 10 was prepared by a synthetic method similar to that in Example 1 and Example 3

[0180] Molecular formula: C 47 H 91N3O5

[0181] Molecular weight: 778.32

[0182] LC-MS: m / z 779 [M+H]

[0183] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.46 (t, 1H, J = 5), 3.80 (m, 2H), 3.62 (m, 2H), 3.40 (m, 2H), 3.35 (m, 2H), 3.02 (t, 1H, J = 3), 2.70 (m, 2H), 2.45 (m, 1H), 2.36 (t, 2H, J = 6), 2.30 (m, 4H), 2.20 (m, 2H), 1.5 - 1.25 (m, 50H), 0.99 (m, 15H).

[0184] Synthesis of Compound 11 in Example 14

[0185]

[0186] A light yellow compound 11 was prepared by a synthetic method similar to that in Example 1 and Example 4

[0187] Molecular formula: C 49 H 95 N3O5

[0188] Molecular weight: 806.32

[0189] LC-MS: m / z 807 [M+H]

[0190] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 3.40 (t, 1H, J = 4.2), 3.35 (m, 4H), 3.30 (m, 3H), 3.10 (m, 1H), 2.65 (m, 2H), 2.45 (m, 8H), 1.52 (m, 2H), 1.44 (m, 6H), 1.38 (m, 6H), 1.29 (m, 6H), 1.26 (m, 40H), 0.90 (m, 15H).

[0191] Synthesis of Compound 12 in Example 15

[0192]

[0193] A light yellow compound 12 was prepared by a synthetic method similar to that in Example 1 and Example 4

[0194] Molecular formula: C 57 H 115 N3O4

[0195] Molecular weight: 906.56

[0196] LC-MS: m / z 907 [M+H]

[0197] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 3.40 (t, 1H, J = 4.2), 3.35 (m, 6H), 3.30 (m, 3H), 3.10 (m, 2H), 2.65 (m, 2H), 2.45 (m, 8H), 1.52 (m, 4H), 1.44 (m, 8H), 1.38 (m, 10H), 1.29 (m, 10H), 1.26 (m, 40H), 0.90 (m, 18H).

[0198] Synthesis of Compound 13 in Example 16

[0199]

[0200] The yellow Compound 13 was prepared by a synthetic method similar to that in Examples 2 and 4

[0201] Molecular formula: C 55 H 107 N3O6

[0202] Molecular weight: 906.53

[0203] LC-MS: m / z 907 [M+H]

[0204] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.76 (t, 1H, J = 6.8 Hz), 4.47 (m, 1H), 3.35 (m, 2H), 3.30 (m, 3H), 3.10 (m, 1H), 2.72 (t, 2H, J = 2.9 Hz), 2.45 (m, 8H), 2.30 (m, 4H), 2.20 (d, 2H, J = 1.3 Hz), 1.64 (m, 4H), 1.50 (m, 8H), 1.38 (m, 4H), 1.29 (m, 10H), 1.26 (m, 36H) 0.90 (m, 18H).

[0205] Synthesis of Compound 14 in Example 17

[0206]

[0207] The yellow Compound 14 was prepared by a synthetic method similar to that in Examples 1, 2 and 4

[0208] Molecular formula: C 56 H 111 N3O5

[0209] Molecular weight: 906.57

[0210] LC-MS: m / z 907 [M+H]

[0211] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.13 (t, 2H, J = 5.2 Hz), 3.39 (t, 1H, J = 3.7 Hz), 3.35 (m, 4H), 3.30 (m, 3H), 3.10 (m, 1H), 2.60 (t, 2H, J = 1.3 Hz), 2.45 (m, 8H), 2.20 (m, 3H), 1.60 (m, 6H), 1.50 (m, 2H), 1.43 (m, 8H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 18H).

[0212] Synthesis of Compound 15 in Example 18

[0213]

[0214] The yellow compound 15 was prepared by a synthetic method similar to that in Examples 1 and 4

[0215] Molecular formula: C50H101N3O4

[0216] Molecular weight: 806.41

[0217] LC-MS: m / z 807 [M+H]

[0218] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.76 (t, 1H, J = 7.9 Hz), 3.35 (m, 2H), 3.30 (m, 3H), 3.10 (m, 1H), 2.70 (t, 2H, J = 3.1 Hz), 2.45 (m, 8H), 2.20 (m, 3H), 1.60 (m, 4H), 1.50 (m, 2H), 1.43 (m, 2H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 18H).

[0219] Synthesis of Compound 16 in Example 19

[0220]

[0221] The yellow compound 16 was prepared by a synthetic method similar to that in Examples 1 and 4

[0222] Molecular formula: C50H101N3O3

[0223] Molecular weight: 792.38

[0224] LC-MS: m / z 800 [M+H]

[0225] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 3.39 (t, 1H, J = 4.3 Hz), 3.35 (m, 4H), 3.30 (m, 3H), 3.10 (m, 1H), 2.70 (t, 2H, J = 3.1 Hz), 2.45 (m, 8H), 2.20 (d, 2H, J = 2.3 Hz), 1.50 (m, 2H), 1.43 (m, 6H), 1.38 (m, 6H), 1.29 (m, 8H), 1.26 (m, 42H), 0.90 (m, 15H).

[0226] Synthesis of Compound 17 in Example 20

[0227]

[0228] The yellow Compound 17 was prepared by a synthetic method similar to that of Examples 1 and 4

[0229] Molecular formula: C49H97N3O4

[0230] Molecular weight: 792.33

[0231] LC-MS: m / z 793 [M+H]

[0232] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.76 (t, 1H, J = 6.8 Hz), 3.35 (m, 2H), 3.30 (m, 3H), 3.10 (m, 1H), 2.70 (t, 2H, J = 3.1 Hz), 2.45 (m, 8H), 2.35 (m, 2H), 2.20 (d, 2H, J = 2.3 Hz), 1.66 (m, 2H), 1.50 (m, 2H), 1.43 (m, 2H), 1.38 (m, 8H), 1.29 (m, 6H), 1.26 (m, 40H), 0.90 (m, 15H).

[0233] Synthesis of Compound 18 in Example 21

[0234]

[0235] The yellow Compound 18 was prepared by a synthetic method similar to that of Examples 1 and 4

[0236] Molecular formula: C 52 H 105 N3O3

[0237] Molecular weight: 820.43

[0238] LC-MS: m / z 821 [M+H]

[0239] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 3.39 (t, 1H, J = 4.3 Hz), 3.35 (m, 4H), 3.30 (m, 3H), 3.10 (m, 1H), 2.70 (t, 2H, J = 3.1 Hz), 2.45 (m, 8H), 2.20 (d, 2H, J = 2.3 Hz), 1.50 (m, 2H), 1.43 (m, 6H), 1.38 (m, 6H), 1.29 (m, 8H), 1.26 (m, 48H), 0.90 (m, 15H).

[0240] Synthesis of Compound 19 in Example 22

[0241]

[0242] The yellow compound 19 was prepared by a synthetic method similar to that of Examples 1 and 4

[0243] Molecular formula: C52H105N3O3

[0244] Molecular weight: 820.43

[0245] LC-MS: m / z 821 [M+H]

[0246] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.76 (t, 1H, J = 6.8 Hz), 3.35 (m, 2H), 3.30 (m, 3H), 3.10 (m, 1H), 2.70 (t, 2H, J = 3.1 Hz), 2.45 (m, 8H), 2.35 (m, 2H), 2.20 (d, 2H, J = 2.3 Hz), 1.66 (m, 2H), 1.50 (m, 2H), 1.43 (m, 2H), 1.38 (m, 8H), 1.29 (m, 6H), 1.26 (m, 46H), 0.90 (m, 15H).

[0247] Synthesis of Compound 20 in Example 23

[0248]

[0249] The yellow compound 20 was prepared by a synthetic method similar to that of Examples 1 and 4

[0250] Molecular formula: C50H98N2O4

[0251] Molecular weight: 791.34

[0252] LC-MS: m / z 792 [M+H]

[0253] 1H NMR (DMSO-d6): δ (ppm) 4.18 (t, 2H, J = 9.2 Hz), 3.40 (t, 1H, J = 4.2), 3.35 (m, 4H), 3.21 (t, 1H, J = 6.2 Hz), 3.10 (m, 1H), 2.93 (m, 2H), 2.65 (d, 2H, J = 2.5 HZ), 2.45 (m, 6H), 1.52 (m, 6H), 1.44 (m, 6H), 1.38 (m, 8H), 1.29 (m, 8H), 1.26 (m, 40H), 0.90 (m, 9H).

[0254] Synthesis of Compound 21 in Example 24

[0255]

[0256]

[0257] The yellow Compound 21 was prepared by a synthetic method similar to that of Examples 1 and 4

[0258] Molecular formula: C49H99N3O5

[0259] Molecular weight: 810.35

[0260] LC-MS: m / z 811 [M+H]

[0261] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.18 (s, 2H), 3.40 (m, 5H,), 3.35 (m, 4H), 3.30 (m, 3H), 3.10 (m, 1H), 2.65 (d, 2H, J = 2.5 HZ), 2.59 (m, 4H), 2.45 (m, 4H), 2.10 (d, 1H, J = 3.3 Hz), 1.85 (d, 1H, J = 3.8 Hz) 1.52 (m, 2H), 1.44 (m, 6H), 1.38 (m, 6H), 1.29 (m, 8H), 1.26 (m, 44H), 0.90 (m, 9H).

[0262] Synthesis of Compound 22 in Example 25

[0263]

[0264] The yellow Compound 22 was prepared by a synthetic method similar to that of Examples 1 and 4

[0265] Molecular formula: C50H99N3O4

[0266] Molecular weight: 806.36

[0267] LC-MS: m / z 807 [M+H]

[0268] 1H NMR (DMSO-d6): δ (ppm) 4.18 (s, 2H), 3.40 (t, 1H, J = 4.1 Hz), 3.35 (m, 4H), 3.30 (m, 1H), 3.18 (t, 1H, J = 3.2 Hz), 3.10 (m, 1H), 2.98 (t, 2H, J = 3.35 Hz), 2.65 (d, 1H, J = 2.5 HZ), 2.36 (d, 1H, J = 2.83 Hz), 2.44 (m, 2H), 2.30 (m, 8H) 2.14 (s, 3H), 1.52 (m, 2H), 1.44 (m, 6H), 1.38 (m, 6H), 1.29 (m, 8H), 1.26 (m, 40H), 0.90 (m, 9H).

[0269] Synthesis of Compound 23 in Example 26

[0270]

[0271] The yellow compound 23 was prepared by a synthetic method similar to that of Examples 1 and 4

[0272] Molecular formula: C49H96N2O5

[0273] Molecular weight: 793.32

[0274] LC-MS: m / z 794 [M+H]

[0275] 1H NMR (DMSO-d6): δ (ppm) 4.18 (s, 2H), 3.58 (m, 4H), 3.40 (t, 1H, J = 4.1 Hz), 3.35 (m, 4H), 3.30 (m, 1H), 3.18 (t, 1H, J = 3.2 Hz), 3.10 (m, 1H), 2.98 (t, 2H, J = 3.35 Hz), 2.65 (d, 1H, J = 2.5 HZ), 2.52 (m, 4H), 2.45 (m, 2H) 2.36 (d, 1H, J = 2.83 Hz), 2.17 (d, 1H, J = 1.82 Hz), 2.03 (d, 1H, J = 1.56 Hz), 1.52 (m, 2H), 1.44 (m, 6H), 1.38 (m, 6H), 1.29 (m, 8H), 1.26 (m, 40H), 0.90 (m, 9H).

[0276] Synthesis of Compound 24 in Example 27

[0277]

[0278] The yellow compound 24 was prepared by a synthetic method similar to that of Example 1 and 4.

[0279] Molecular formula: C49H96N2O4

[0280] Molecular weight: 777.32

[0281] LC-MS: m / z 778 [M+H]

[0282] 1H NMR (DMSO-d6): δ (ppm) 4.23 (s, 2H), 3.40 (t, 1H, J = 4.1 Hz), 3.35 (m, 4H), 3.30 (m, 1H), 3.18 (t, 1H, J = 3.2 Hz), 3.10 (m, 1H), 2.98 (t, 2H, J = 3.35 Hz), 2.65 (d, 1H, J = 2.5 HZ), 2.52 (m, 4H), 2.36 (d, 1H, J = 2.83 Hz), 2.17 (d, 1H, J = 1.87 Hz), 2.00 (d, 1H, J = 1.72 Hz), 1.70 (m, 4H), 1.52 (m, 2H), 1.44 (m, 6H), 1.38 (m, 6H), 1.29 (m, 8H), 1.26 (m, 40H), 0.90 (m, 9H).

[0283] Synthesis of Compound 25 in Example 28

[0284]

[0285] The yellow compound 25 was prepared by a synthetic method similar to that of Example 1 and 4.

[0286] Molecular formula: C48H96N2O4

[0287] Molecular weight: 765.31

[0288] LC-MS: m / z 766 [M+H]

[0289] 1H NMR (DMSO-d6): δ (ppm) 4.10 (s, 2H), 3.40 (t, 1H, J = 4.1 Hz), 3.35 (m, 4H), 3.30 (m, 1H), 3.18 (t, 1H, J = 3.2 Hz), 3.10 (m, 1H), 2.65 (d, 1H, J = 2.5 HZ), 2.36 (d, 1H, J = 2.83 Hz), 2.32 (m, 2H), 2.17 (m, 6H),, 1.70 (m, 2H), 1.52 (m, 2H), 1.44 (m, 6H), 1.38 (m, 6H), 1.29 (m, 8H), 1.26 (m, 40H) 0.90 (m, 9H).

[0290] Synthesis of Compound 26 in Example 29

[0291]

[0292] The yellow compound 26 was prepared by a synthetic method similar to that of Examples 1 and 4

[0293] Molecular formula: C57H115N3O6

[0294] Molecular weight: 938.56

[0295] LC-MS: m / z 939 [M + H]

[0296] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.52 (s, 2H), 3.45 (m, 4H), 3.40 (t, 1H, J = 4.1 Hz), 3.35 (m, 6H), 3.30 (m, 3H), 3.10 (m, 2H), 2.65 (d, 1H, J = 2.5 HZ), 2.58 (m, 4H), 2.46 (m, 4H), 2.36 (d, 1H, J = 2.83 Hz), 2.32 (m, 2H), 2.10 (d, 1H, J = 3.8 Hz), 1.95 (d, 1H, J = 3.01 Hz), 1.52 (m, 4H), 1.44 (m, 8H), 1.38 (m, 8H), 1.29 (m, 10H), 1.26 (m, 40H), 0.90 (m, 12H).

[0297] Synthesis of Compound 27 in Example 30

[0298]

[0299] The yellow compound 27 was prepared by a synthetic method similar to that of Examples 1 and 4

[0300] Molecular formula: C56H112N2O5

[0301] Molecular weight: 893.52

[0302] LC-MS: m / z 894 [M+H]

[0303] 1H NMR (DMSO-d6): δ (ppm) 4.18 (t, 2H, J = 2.3 Hz), 3.45 (t, 1H, J = 3.3 Hz), 3.35 (m, 6H), 3.30 (m, 3H), 3.10 (m, 2H), 2.65 (d, 1H, J = 2.5 HZ), 2.38 (m, 2H), 2.36 (d, 1H, J = 2.83 Hz), 2.15 (s, 6H), 2.10 (d, 1H, J = 3.8 Hz), 1.95 (d, 1H, J = 3.01 Hz), 1.68 (m, 2H), 1.52 (m, 4H), 1.44 (m, 8H), 1.38 (m, 8H), 1.29 (m, 10H), 1.26 (m, 40H) 0.90 (m, 12H).

[0304] Synthesis of Compound 28 in Example 31

[0305]

[0306] The yellow Compound 28 was prepared by a synthetic method similar to that of Examples 1 and 4

[0307] Molecular formula: C58H114N2O5

[0308] Molecular weight: 919.56

[0309] LC-MS: m / z 920 [M+H]

[0310] 1H NMR (DMSO-d6): δ (ppm) 4.18 (t, 2H, J = 2.3 Hz), 3.45 (t, 1H, J = 3.3 Hz), 3.35 (m, 6H), 3.30 (m, 3H), 3.10 (m, 2H), 2.97 (m, 2H), 2.65 (d, 1H, J = 2.5 HZ), 2.46 (m, 4H), 2.38 (m, 2H), 2.36 (d, 1H, J = 2.83 Hz), 2.10 (d, 1H, J = 3.8 Hz), 1.95 (d, 1H, J = 3.01 Hz), 1.52 (m, 4H), 1.47 (m, 4H), 1.44 (m, 8H), 1.38 (m, 10H), 1.29 (m, 10H), 1.26 (m, 40H) 0.90 (m, 12H).

[0311] Synthesis of Compound 29 in Example 32

[0312]

[0313] The yellow compound 29 was prepared by a synthetic method similar to that of Examples 1 and 4.

[0314] Molecular formula: C58H114N2O6

[0315] Molecular weight: 935.56

[0316] LC-MS: m / z 936 [M+H]

[0317] 1H NMR (DMSO-d6): δ (ppm) 5.52 (s, 1H), 4.18 (t, 2H, J = 2.3 Hz), 4.01 (m, 1H), 3.45 (t, 1H, J = 3.3 Hz), 3.35 (m, 6H), 3.20 (t, 1H, J = 6.7 Hz), 3.10 (m, 2H), 2.97 (m, 2H), 2.65 (d, 1H, J = 2.5 HZ), 2.45 - 2.53 (m, 6H), 2.36 (d, 1H, J = 2.83 Hz), 2.10 (d, 1H, J = 3.8 Hz), 1.99 (d, 1H, J = 3.6 Hz), 1.82 (m, 4H), 1.52 (m, 4H), 1.44 (m, 8H), 1.38 (m, 10H), 1.29 (m, 10H), 1.26 (m, 40H) 0.90 (m, 12H).

[0318] Synthesis of Compound 30 in Example 33

[0319]

[0320] The yellow compound 30 was prepared by a synthetic method similar to that of Examples 1 and 4.

[0321] Molecular formula: C57H112N2O5

[0322] Molecular weight: 905.53

[0323] LC-MS: m / z 906 [M+H]

[0324] 1H NMR (DMSO-d6): δ (ppm) 4.18 (t, 2H, J = 2.3 Hz), 3.45 (t, 1H, J = 3.3 Hz), 3.35 (m, 6H), 3.20 (t, 1H, J = 6.7 Hz), 3.10 (m, 2H), 2.97 (m, 2H), 2.65 (d, 1H, J = 2.5 HZ), 2.53 (m, 4H), 2.45 (m, 2H), 2.36 (d, 1H, J = 2.83 Hz), 2.10 (d, 1H, J = 3.8 Hz), 1.99 (d, 1H, J = 3.6 Hz), 1.69 (m, 4H), 1.52 (m, 4H), 1.44 (m, 8H), 1.38 (m, 10H), 1.29 (m, 10H), 1.26 (m, 40H) 0.90 (m, 12H).

[0325] Synthesis of Compound 31 in Example 34

[0326]

[0327] A yellow compound 31 was prepared by a synthesis method similar to that of Examples 1 and 4

[0328] Molecular formula: C58H115N3O5

[0329] Molecular weight: 934.57

[0330] LC-MS: m / z 935 [M + H]

[0331] 1H NMR (DMSO-d6): δ (ppm) 4.18 (t, 2H, J = 2.3 Hz), 3.45 (t, 1H, J = 3.3 Hz), 3.35 (m, 6H), 3.20 (t, 1H, J = 6.7 Hz), 3.10 (m, 2H), 2.97 (m, 2H), 2.65 (d, 1H, J = 2.5 HZ), 2.53 (m, 4H), 2.45 (m, 2H), 2.36 (d, 1H, J = 2.83 Hz), 2.30 (m, 8H), 2.20 (d, 1H, J = 3.8 Hz), 2.15 (s, 3H), 1.99 (d, 1H, J = 3.6 Hz), 1.52 (m, 4H), 1.44 (m, 8H), 1.38 (m, 10H), 1.29 (m, 10H), 1.26 (m, 40H), 0.90 (m, 12H).

[0332] Synthesis of Compound 32 in Example 35

[0333]

[0334] A yellow compound 32 was prepared by a synthesis method similar to that of Examples 1 and 4

[0335] Molecular formula: C50H99N3O6

[0336] Molecular weight: 838.41

[0337] LC-MS: m / z 839 [M+H]

[0338] 1H NMR (DMSO-d6): δ (ppm) 8.06 (s, 1H), 4.76 (t, 1H, J = 7.9 Hz), 4.52 (s, 2H), 3.48 (m, 4H), 3.35 (m, 2H), 3.30 (m, 3H), 3.10 (m, 1H), 2.70 (d, 2H, J = 3.1 Hz), 2.62 (m, 4H), 2.45 (m, 4H), 2.20 (m, 3H), 1.60 (m, 4H), 1.50 (m, 2H), 1.43 (m, 2H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0339] Synthesis of Compound 33 in Example 36

[0340]

[0341] The yellow Compound 33 was prepared by a synthetic method similar to that of Examples 1 and 4

[0342] Molecular formula: C49H96N2O5

[0343] Molecular weight: 793.37

[0344] LC-MS: m / z 794 [M+H]

[0345] 1H NMR (DMSO-d6): δ (ppm) 4.76 (t, 1H, J = 7.9 Hz), 4.12 (m, 2H), 3.35 (m, 2H), 3.20 (m, 1H), 3.10 (m, 1H), 2.70 (d, 2H, J = 3.1 Hz), 2.45 (m, 4H), 2.20 (m, 7H), 2.31 (m, 2H), 1.70 (m, 6H), 1.50 (m, 6H), 1.43 (m, 2H), 1.38 (m, 8H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0346] Synthesis of Compound 34 in Example 37

[0347]

[0348] The yellow compound 34 was prepared by a synthesis method similar to that of Example 1 and 4.

[0349] Molecular formula: C48H94N2O5

[0350] Molecular weight: 793.37

[0351] LC-MS: m / z 794 [M+H]

[0352] 1H NMR (DMSO-d6): δ (ppm) 4.76 (t, 1H, J = 7.9 Hz), 4.12 (m, 2H), 3.35 (m, 2H), 3.20 (m, 1H), 3.10 (m, 1H), 2.70 (d, 2H, J = 3.1 Hz), 2.45 (m, 4H), 2.20 (m, 7H), 2.31 (m, 2H), 1.70 (m, 6H), 1.50 (m, 6H), 1.43 (m, 2H), 1.38 (m, 8H), 1.29 (m, 8H), 1.26 (m, 36H), 0.90 (m, 12H).

[0353] Synthesis of Compound 35 in Example 38

[0354]

[0355] The yellow compound 35 was prepared by a synthesis method similar to that of Example 1 and 4.

[0356] Molecular formula: C44H86N2O5

[0357] Molecular weight: 723.18

[0358] LC-MS: m / z 724 [M+H]

[0359] 1H NMR (DMSO-d6): δ (ppm) 4.76 (t, 1H, J = 7.9 Hz), 4.12 (m, 2H), 3.35 (m, 2H), 3.20 (m, 1H), 3.10 (m, 1H), 2.70 (d, 2H, J = 3.1 Hz), 2.45 (m, 4H), 2.20 (m, 7H), 2.31 (m, 2H), 1.70 (m, 6H), 1.50 (m, 6H), 1.43 (m, 2H), 1.38 (m, 8H), 1.29 (m, 10H), 1.26 (m, 34H), 0.90 (m, 12H).

[0360] Synthesis of Compound 36 in Example 39

[0361]

[0362] The yellow compound 36 was prepared by a synthetic method similar to that of Example 1 and 4

[0363] Molecular formula: C51H88N2O5

[0364] Molecular weight: 819.41

[0365] LC-MS: m / z 820 [M+H]

[0366] 1H NMR (DMSO-d6): δ (ppm) 4.76 (t, 1H, J = 7.9 Hz), 4.12 (m, 2H), 3.35 (m, 2H), 3.20 (m, 1H), 3.10 (m, 1H), 2.97 (m, 2H), 2.70 (d, 2H, J = 3.1 Hz), 2.45 (m, 4H), 2.20 (m, 1H), 2.31 (m, 2H), 1.50 (m, 2H), 1.43 (m, 2H), 1.38 (m, 8H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0367] Synthesis of Compound 37 in Example 40

[0368]

[0369] The yellow compound 37 was prepared by a synthetic method similar to that of Example 1 and 4

[0370] Molecular formula: C50H96N2O6

[0371] Molecular weight: 821.38

[0372] LC-MS: m / z 822 [M+H]

[0373] 1H NMR (DMSO-d6): δ (ppm) 4.76 (t, 1H, J = 7.9 Hz), 4.12 (m, 2H), 3.59 (m, 4H), 3.35 (m, 2H), 3.20 (m, 1H), 3.10 (m, 1H), 2.97 (m, 2H), 2.70 (d, 2H, J = 3.1 Hz), 2.45 (m, 4H), 2.20 (m, 1H), 2.31 (m, 2H), 1.50 (m, 2H), 1.43 (m, 2H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0374] Synthesis of Compound 38 in Example 41

[0375]

[0376] The yellow compound 38 was prepared by a synthetic method similar to that of Example 1 and 4

[0377] Molecular formula: C49H94N2O6

[0378] Molecular weight: 835.41

[0379] LC-MS: m / z 836 [M+H]

[0380] 1H NMR (DMSO-d6): δ (ppm) 4.76 (t, 1H, J = 7.9 Hz), 4.49 (s, 1H), 4.20 (m, 2H), 3.59 (m, 1H), 3.35 (m, 2H), 3.20 (m, 1H), 3.10 (m, 1H), 2.97 (m, 2H), 2.70 (d, 2H, J = 3.1 Hz), 2.45 (m, 6H), 2.20 (m, 1H), 2.31 (m, 2H), 1.77 (m, 4H), 1.50 (m, 2H), 1.43 (m, 2H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0381] Synthesis of Compound 39 in Example 42

[0382]

[0383] The yellow compound 39 was prepared by a synthetic method similar to that of Example 1 and 4

[0384] Molecular formula: C50H96N2O5

[0385] Molecular weight: 805.38

[0386] LC-MS: m / z 806 [M+H]

[0387] 1H NMR (DMSO-d6): δ (ppm) 4.76 (t, 1H, J = 7.9 Hz), 4.20 (m, 2H), 3.35 (m, 2H), 3.20 (m, 1H), 3.10 (m, 1H), 2.97 (m, 2H), 2.70 (d, 2H, J = 3.1 Hz), 2.45 (m, 6H), 2.20 (m, 1H), 2.31 (m, 2H), 1.71 (m, 4H), 1.50 (m, 2H), 1.43 (m, 2H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0388] Synthesis of Compound 40 in Example 43

[0389]

[0390] The yellow compound 40 was prepared by a synthesis method similar to that of Examples 1 and 4

[0391] Molecular formula: C51H99N3O5

[0392] Molecular weight: 834.42

[0393] LC-MS: m / z 835 [M+H]

[0394] 1H NMR (DMSO-d6): δ (ppm) 4.76 (t, 1H, J = 7.9 Hz), 4.20 (m, 2H), 3.35 (m, 2H), 3.20 (m, 1H), 3.10 (m, 1H), 2.97 (m, 2H), 2.70 (d, 2H, J = 3.1 Hz), 2.45 (m, 2H), 2.31 (m, 2H), 2.29 (m, 8H), 2.18 (m, 1H), 2.14 (s, 3H), 1.50 (m, 2H), 1.43 (m, 2H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0395] Synthesis of Compound 41 in Example 44

[0396]

[0397] The yellow compound 41 was prepared by a synthesis method similar to that of Examples 1, 2 and 4

[0398] Molecular formula: C56H111N3O7

[0399] Molecular weight: 938.57

[0400] LC-MS: m / z 938 [M+H]

[0401] 1H NMR (DMSO-d6): δ (ppm) 4.20 (s, 2H), 4.18 (m, 2H), 4.15 (m, 2H), 3.46 (t, 1H, J = 3.2 Hz), 3.43 (m, 4H), 3.35 (m, 4H), 3.20 (m, 3H), 3.10 (m, 1H), 2.62 (d, 2H, J = 2.1 Hz), 2.57 (m, 4H), 2.45 (m, 4H), 2.18 (m, 1H), 2.14 (d, 2H, J = 3.5 Hz), 1.62 (m, 6H), 1.50 (m, 2H), 1.43 (m, 8H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0402] Synthesis of Compound 42 in Example 45

[0403]

[0404] The yellow Compound 42 was prepared by a synthetic method similar to that of Examples 1, 2, and 4

[0405] Molecular formula: C57H110N2O7

[0406] Molecular weight: 935.51

[0407] LC-MS: m / z 936 [M+H]

[0408] 1H NMR (DMSO-d6): δ (ppm) 4.72 (s, 1H), 4.18 (m, 2H), 4.15 (m, 2H), 3.46 (t, 1H, J = 3.2 Hz), 3.72 (m, 1H), 3.35 (m, 4H), 3.20 (m, 3H), 3.10 (m, 1H), 2.62 (d, 2H, J = 2.1 Hz), 2.57 (m, 4H), 2.45 (m, 6H), 2.18 (m, 1H), 2.14 (d, 2H, J = 3.5 Hz), 1.82 (m, 4H), 1.62 (m, 6H), 1.50 (m, 2H), 1.43 (m, 8H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H) 0.90 (m, 12H).

[0409] Synthesis of Compound 43 in Example 46

[0410]

[0411] The yellow Compound 43 was prepared by a synthetic method similar to that of Examples 1, 2, and 4

[0412] Molecular formula: C57H111N3O6

[0413] Molecular weight: 934.58

[0414] LC-MS: m / z 963 [M+H]

[0415] 1H NMR (DMSO-d6): δ (ppm) 4.33 (m, 2H), 4.15 (m, 2H), 3.46 (t, 1H, J = 3.2 Hz), 3.35 (m, 4H), 3.20 (m, 3H), 3.10 (m, 1H), 2.97 (m, 2H), 2.62 (d, 2H, J = 2.1 Hz), 2.45 (m, 2H), 2.29 (m, 8H), 2.18 (m, 4H), 2.14 (d, 2H, J = 3.5 Hz), 1.62 (m, 6H), 1.50 (m, 2H), 1.43 (m, 8H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0416] Synthesis of Example 47 Compound 44

[0417]

[0418] The yellow compound 44 was prepared by a synthetic method similar to that of Examples 1, 2, and 4

[0419] Molecular formula: C57H110N2O6

[0420] Molecular weight: 919.57

[0421] LC-MS: m / z 920 [M + H]

[0422] 1H NMR (DMSO-d6): δ (ppm) 4.33 (m, 2H), 4.15 (m, 2H), 3.46 (t, 1H, J = 3.2 Hz), 3.35 (m, 4H), 3.20 (m, 3H), 3.10 (m, 1H), 2.97 (m, 2H), 2.62 (d, 2H, J = 2.1 Hz), 2.45 (m, 6H), 2.18 (m, 4H), 2.14 (d, 2H, J = 3.5 Hz), 1.62 (m, 6H), 1.50 (m, 6H), 1.43 (m, 8H), 1.38 (m, 8H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0423] Synthesis of Example 48 Compound 45

[0424]

[0425] The yellow compound 45 was prepared by a synthetic method similar to that of Examples 1, 2, and 4

[0426] Molecular formula: C56H108N2O6

[0427] Molecular weight: 905.54

[0428] LC-MS: m / z 906 [M+H]

[0429] 1H NMR (DMSO-d6): δ (ppm) 4.33 (m, 2H), 4.15 (m, 2H), 3.46 (t, 1H, J = 3.2 Hz), 3.35 (m, 4H), 3.20 (m, 3H), 3.10 (m, 1H), 2.97 (m, 2H), 2.62 (d, 2H, J = 2.1 Hz), 2.45 (m, 6H), 2.14 (d, 2H, J = 3.5 Hz), 1.62 (m, 10H), 1.50 (m, 2H), 1.43 (m, 8H), 1.38 (m, 6H), 1.29 (m, 10H), 1.26 (m, 36H), 0.90 (m, 12H).

[0430] Synthesis of Compound 46 in Example 49

[0431]

[0432] The yellow compound 46 was prepared by a synthetic method similar to that of Examples 1 and 3

[0433] Molecular formula: C57H104N2O10

[0434] Molecular weight: 977.46

[0435] LC-MS: m / z 978 [M+H]

[0436] 1H NMR (DMSO-d6): δ (ppm) 5.14 (t, 1H, J = 9.55 Hz), 4.47 (m, 2H), 4.29 (t, 1H, J = 8.96 Hz), 4.18 (m, 2H), 3.83 (d, 2H, J = 5.67 Hz), 3.57 (m, 4H), 2.97 (m, 2H), 2.69 (d, 2H, J = 4.23 Hz), 2.50 (m, 4H), 2.35 (m, 6H), 2.05 (m, 2H) 1.64 (m, 6H), 1.26 - 1.50 (m, 58H), 0.90 (m, 12H).

[0437] Synthesis of Compound 47 in Example 50

[0438]

[0439] The yellow compound 47 was prepared by a synthetic method similar to that of Examples 1 and 3

[0440] Molecular formula: C58H107N3O9

[0441] Molecular weight: 990.51

[0442] LC-MS: m / z 991 [M+H]

[0443] 1H NMR (DMSO-d6): δ (ppm) 5.14 (t, 1H, J = 9.55 Hz), 4.47 (m, 2H), 4.29 (t, 1H, J = 8.96 Hz), 4.18 (m, 2H), 3.83 (d, 2H, J = 5.67 Hz), 2.97 (m, 2H), 2.69 (d, 2H, J = 4.23 Hz), 2.35 (m, 6H), 2.29 (m, 8H), 2.14 (s, 3H), 2.05 (m, 2H), 1.64 (m, 6H), 1.26 - 1.50 (m, 58H), 0.90 (m, 12H).

[0444] Synthesis of Compound 48 in Example 51

[0445]

[0446] A yellow compound 48 was prepared by a synthetic method similar to that of Examples 1 and 3

[0447] Molecular formula: C57H106N2O11

[0448] Molecular weight: 995.48

[0449] LC-MS: m / z 996 [M+H]

[0450] 1H NMR (DMSO-d6): δ (ppm) 5.14 (t, 1H, J = 9.55 Hz), 4.47 (m, 2H), 4.29 (t, 1H, J = 8.96 Hz), 4.18 (m, 2H), 4.16 (s, 2H), 3.83 (d, 2H, J = 5.67 Hz), 3.42 (m, 4H), 2.97 (m, 2H), 2.69 (d, 2H, J = 4.23 Hz), 2.57 (m, 4H), 2.35 (m, 6H), 2.05 (m, 2H), 1.64 (m, 6H), 1.26 - 1.50 (m, 58H), 0.90 (m, 12H).

[0451] Synthesis of Compound 49 in Example 52

[0452]

[0453] A yellow compound 49 was prepared by a synthetic method similar to that of Examples 1 and 3

[0454] Molecular formula: C58H106N2O10

[0455] Molecular weight: 991.49

[0456] LC-MS: m / z 936 [M+H]

[0457] 1H NMR (DMSO-d6): δ (ppm) 5.14 (t, 1H, J = 9.55 Hz), 4.47 (m, 2H), 4.29 (t, 1H, J = 8.96 Hz), 4.18 (m, 2H), 4.49 (s, 2H), 3.83 (d, 2H, J = 5.67 Hz), 3.60 (m, 1H), 2.97 (m, 2H), 2.69 (d, 2H, J = 4.23 Hz), 2.57 (m, 4H), 2.35 (m, 6H), 2.05 (m, 2H), 1.77 (m, 4H), 1.64 (m, 6H), 1.26 - 1.50 (m, 58H), 0.90 (m, 12H).

[0458] Synthesis of Compound 50 in Example 53

[0459]

[0460] The yellow Compound 50 was prepared by a synthetic method similar to that of Examples 1 and 3

[0461] Molecular formula: C44H83N3O5

[0462] Molecular weight: 734.27

[0463] LC-MS: m / z 735 [M+H]

[0464] 1H NMR (DMSO-d6): δ (ppm) 5.14 (t, 1H, J = 9.55 Hz), 4.29 (t, 1H, J = 8.96 Hz), 4.18 (m, 2H), 3.83 (d, 2H, J = 5.67 Hz), 2.97 (m, 2H), 2.69 (d, 2H, J = 4.23 Hz), 2.29 (m, 8H), 2.27 (m, 1H), 2.14 (m, 3H), 2.13 (m, 1H), 1.60 (m, 4H), 1.26 - 1.50 (m, 48H), 0.90 (m, 12H).

[0465] Synthesis of Compound 51 in Example 54

[0466]

[0467] The yellow Compound 51 was prepared by a synthetic method similar to that of Example 1

[0468] Molecular formula: C43H82N2O7

[0469] Molecular weight: 739.24

[0470] LC-MS: m / z 7940 [M+H]

[0471] 1H NMR (DMSO-d6): δ (ppm) 5.14 (t, 1H, J = 9.55 Hz), 4.29 (t, 1H, J = 8.96 Hz), 4.18 (m, 2H), 4.16 (s, 2H), 3.83 (d, 2H, J = 5.67 Hz), 3.42 (m, 2H), 2.97 (m, 2H), 2.69 (d, 2H, J = 4.23 Hz), 2.57 (m, 4H),, 2.27 (m, 1H), 2.13 (m, 1H), 1.60 (m, 4H), 1.26 - 1.50 (m, 48H), 0.90 (m, 12H).

[0472] Synthesis of Compound 52 in Example 55

[0473]

[0474] The yellow Compound 52 was prepared by a synthesis method similar to that of Example 1

[0475] Molecular formula: C44H82N2O6

[0476] Molecular weight: 735.26

[0477] LC-MS: m / z 736 [M+H]

[0478] 1H NMR (DMSO-d6): δ (ppm) 5.14 (t, 1H, J = 9.55 Hz), 4.49 (s, 1H), 4.29 (t, 1H, J = 8.96 Hz), 4.18 (m, 2H), 3.83 (d, 2H, J = 5.67 Hz), 3.60 (m, 1H), 2.97 (m, 2H), 2.69 (d, 2H, J = 4.23 Hz), 2.51 (m, 4H),, 2.27 (m, 1H), 2.13 (m, 1H), 1.77 (m, 4H), 1.60 (m, 4H), 1.26 - 1.50 (m, 48H), 0.90 (m, 12H).

[0479] Synthesis of Compound 53 in Example 56

[0480]

[0481] The yellow Compound 53 was prepared by a synthesis method similar to that of Examples 1 and 2

[0482] Molecular formula: C55H107N3O8

[0483] Molecular weight: 938.47

[0484] LC-MS: m / z 995 [M+H]

[0485] 1H NMR (DMSO-d6): δ (ppm) 8.01 (s, 1H), 4.16 (s, 2H), 4.13 (m, 4H), 3.42 (m, 2H), 3.39 (t, 1H, J = 3.2 Hz), 3.35 (m, 2H), 3.30 (m, 3H), 2.69 (d, 2H, J = 4.23 Hz), 2.57 (m, 4H), 2.43 (m, 4H), 2.13 (m, 2H), 1.60 (m, 12H), 1.26 - 1.50 (m, 52H), 0.90 (m, 12H).

[0486] Synthesis of Compound 54 in Example 57

[0487]

[0488] A yellow compound 54 was prepared by a synthetic method similar to that of Examples 1 and 2

[0489] Molecular formula: C55H104N2O8

[0490] Molecular weight: 921.44

[0491] LC-MS: m / z 922 [M+H]

[0492] 1H NMR (DMSO-d6): δ (ppm) 4.18 (m, 2H), 4.13 (m, 4H), 3.57 (m, 4H), 3.39 (t, 1H, J = 3.2 Hz), 3.35 (m, 2H), 3.18 (t, 1H, J = 3.51 Hz), 2.69 (d, 2H, J = 4.23 Hz), 2.50 (m, 4H), 2.43 (m, 2H), 2.13 (m, 2H), 1.60 (m, 12H), 1.26 - 1.50 (m, 52H), 0.90 (m, 12H).

[0493] Synthesis of Compound 55 in Example 58

[0494]

[0495] A yellow compound 55 was prepared by a synthetic method similar to that of Examples 1 and 2

[0496] Molecular formula: C56H107N3O7

[0497] Molecular weight: 934.49

[0498] LC-MS: m / z 935 [M+H]

[0499] 1H NMR (DMSO-d6): δ (ppm) 4.18 (m, 2H), 4.13 (m, 4H), 3.57 (m, 4H), 3.39 (t, 1H, J = 3.2 Hz), 3.35 (m, 2H), 3.18 (t, 1H, J = 3.51 Hz), 2.97 (m, 2H), 2.69 (d, 2H, J = 4.23 Hz), 2.29 (m, 8H), 2.14 (s, 3H), 2.13 (m, 2H), 1.60 (m, 12H), 1.26 - 1.50 (m, 52H), 0.90 (m, 12H).

[0500] Synthesis of Compound 56 in Example 59

[0501]

[0502] The yellow compound 56 was prepared by a synthetic method similar to that of Examples 1 and 2

[0503] Molecular formula: C56H106N2O8

[0504] Molecular weight: 935.47

[0505] LC-MS: m / z 936 [M+H]

[0506] 1H NMR (DMSO-d6): δ (ppm) 4.49 (s, 1H) 4.18 (m, 2H), 4.13 (m, 4H),, 3.60 (m, 1H), 3.39 (t, 1H, J = 3.2 Hz), 3.35 (m, 2H), 3.18 (t, 1H, J = 3.51 Hz), 2.97 (m, 2H), 2.69 (d, 2H, J = 4.23 Hz), 2.52 (m, 4H), 2.13 (m, 2H), 1.77 (m, 4H), 1.60 (m, 12H), 1.26 - 1.50 (m, 52H), 0.90 (m, 12H).

[0507] Synthesis of Compound 57 in Example 60

[0508]

[0509] The yellow compound 57 was prepared by a synthetic method similar to that of Examples 1 and 2

[0510] Molecular formula: C57H106N2O9

[0511] Molecular weight: 963.48

[0512] LC-MS: m / z 964 [M+H]

[0513] 1H NMR (DMSO-d6): δ (ppm), 4.76 (d, 1H, J = 8.22 Hz), 4.49 (s, 1H), 4.47 (m, 1H), 4.18 (m, 2H), 4.13 (m, 2H), 3.60 (m, 1H), 3.39 (t, 1H, J = 3.2 Hz), 3.35 (m, 2H), 3.18 (t, 1H, J = 3.51 Hz), 2.97 (m, 2H), 2.71 (d, 2H, J = 4.23 Hz), 2.52 (m, 4H), 2.32 (m, 4H), 2.13 (m, 1H), 1.77 (m, 4H), 1.64 (m, 4H), 1.60 (m, 6H), 1.26 - 1.50 (m, 50H), 0.90 (m, 12H).

[0514] Synthesis of Compound 58 in Example 61

[0515]

[0516] A yellow compound 58 was prepared by a synthesis method similar to that of Examples 1 and 2

[0517] Molecular formula: C57H106N3O8

[0518] Molecular weight: 963.48

[0519] LC-MS: m / z 936 [M+H]

[0520] 1H NMR (DMSO-d6): δ (ppm), 4.76 (d, 1H, J = 8.22 Hz),, 4.47 (m, 1H), 4.18 (m, 2H), 4.13 (m, 2H), 3.39 (t, 1H, J = 3.2 Hz), 3.35 (m, 2H), 3.18 (t, 1H, J = 3.51 Hz), 2.97 (m, 2H), 2.71 (d, 2H, J = 4.23 Hz), 2.32 (m, 4H), 2.29 (m, 8H), 2.14 (s, 3H), 2.13 (m, 1H), 1.64 (m, 4H), 1.60 (m, 6H), 1.26 - 1.50 (m, 50H), 0.90 (m, 12H).

[0521] Synthesis of Compound 59 in Example 62

[0522]

[0523] A yellow compound 59 was prepared by a synthesis method similar to that of Examples 1 and 2

[0524] Molecular formula: C56H104N2O9

[0525] Molecular weight: 949.45

[0526] LC-MS: m / z 950 [M+H]

[0527] 1H NMR (DMSO-d6): δ (ppm), 4.76 (d, 1H, J = 8.22 Hz),, 4.47 (m, 1H), 4.18 (m, 2H), 4.13 (m, 2H), 3.57 (m, 4H), 3.39 (t, 1H, J = 3.2 Hz), 3.35 (m, 2H), 3.18 (t, 1H, J = 3.51 Hz), 2.97 (m, 2H), 2.71 (d, 2H, J = 4.23 Hz), 2.50 (m, 4H), 2.32 (m, 4H), 2.14 (s, 3H), 2.13 (m, 1H), 1.64 (m, 4H), 1.60 (m, 6H), 1.26 - 1.50 (m, 50H), 0.90 (m, 12H).

[0528] Synthesis of Compound 60 in Example 63

[0529]

[0530] The yellow compound 60 was prepared by a synthetic method similar to that of Examples 1 and 2

[0531] Molecular formula: C52H107N3O9

[0532] Molecular weight: 966.54

[0533] LC-MS: m / z 967 [M+H]

[0534] 1H NMR (DMSO-d6): δ (ppm), 4.76 (d, 1H, J = 8.22 Hz),, 4.47 (m, 1H), 4.16 (s, 2H), 4.13 (m, 2H), 3.42 (m, 2H), 3.39 (t, 1H, J = 3.2 Hz), 3.35 (m, 2H), 3.30 (m, 2H), 3.18 (t, 1H, J = 3.51 Hz), 2.71 (d, 2H, J = 4.23 Hz), 2.46 (m, 2H), 2.32 (m, 4H), 2.14 (s, 3H), 2.13 (m, 1H), 1.64 (m, 4H), 1.60 (m, 6H), 1.26 - 1.50 (m, 50H), 0.90 (m, 12H).

[0535] Example 64

[0536] In Vivo Evaluation of Luciferase mRNA Using Lipid Nanoparticle Compositions

[0537] 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 mRNA to 0.15 mg / mL in 10 mL to 50 mL of citrate buffer (pH = 4). Using a syringe pump, mix the ethanol solution of lipid with the aqueous solution of 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 determined 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.

[0538] Conduct studies 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 Council. Systemically administer different doses of mRNA lipid nanoparticles via tail vein injection and euthanize the animals at specific time points after administration (e.g., 5 hours). Collect the liver and spleen in pre-weighed tubes, determine the weight, immediately snap-freeze in liquid nitrogen, and store at -80 °C until used for analysis.

[0539] For the liver, approximately 50 mg was cut for analysis in 2 mL FastPrep tubes (MP Biomedicals, Solon OH). 1 / 4" ceramic beads (MP Biomedicals) were 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 at 2 x 6.0 m / s for 15 seconds using a FastPrep24 instrument (MP Biomedicals). The homogenate was incubated for 5 minutes at room temperature, 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 assayed was determined by 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 luciferase, a standard curve was generated using QuantiLum recombinant luciferase (Promega).

[0540] FLuc mRNA (L - 6107) from Trilink Biotechnologies will express 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.

[0541] Example 65

[0542] Determination of the pKa of the formulated lipid

[0543] 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 analysis based on the fluorescence of 2-(p-toluidino)-6-naphthalene sulfonic acid (TNS). As described in Example 64, 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 gives half-maximal fluorescence intensity.

[0544] Example 66

[0545] The efficacy of lipid nanoparticle formulations containing various cationic lipids was determined using a rodent model of in vivo luciferase mRNA expression.

[0546] For comparison purposes, these lipids were also used to formulate lipid nanoparticles containing FLuc mRNA (L-6107) using an ordered mixing method as described in Example 64. 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, average PEG molecular weight of 2000). As described in Example 64, 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 64. The results of Examples 64 and 65 are shown in Table 2.

[0547] Table 2 Comparison of lipids showing activity with mRNA

[0548]

[0549]

[0550]

[0551]

[0552]

[0553]

[0554]

[0555]

[0556]

[0557]

[0558]

[0559]

[0560]

[0561]

[0562]

[0563]

[0564] 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-described 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.

[0565] The above-described embodiments only represent several implementation manners of the present invention, and their descriptions are relatively specific and detailed, but they 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 29 ; Compound 32: ; Compound 33 ; Compound 35 ; Compound 37 ; Compound 39: ; Compound 40 ; Compound 41: ; Compound 42 ; Compound 45 ; Compound 46: ; Compound 47: .

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 drug or a vaccine for treating a disease selected from cancer, proliferative diseases, neurodegenerative diseases, or metabolic diseases.

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