Phospholipid drug conjugate, and preparation method and use therefor

Abstract

Disclosed in the present invention are a phospholipid drug conjugate and a preparation method and use therefor. Disclosed are the phospholipid drug conjugate as shown in formula (I), or a stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate thereof. The phospholipid drug conjugate disclosed by the invention has good water solubility, can self-assemble to form a nanostructure, and has an excellent tumor inhibition effect.

Description

A phospholipid drug conjugate and its preparation method and use

[0001] This application claims the benefit of Chinese Patent Application No. 2023112617069, filed September 27, 2023. This application incorporates the entirety of the aforementioned Chinese Patent Application. Technical Field

[0002] The present invention belongs to the field of drug delivery, and in particular relates to a phospholipid drug conjugate and a preparation method and application thereof. Background Art

[0003] The incidence and mortality of cancer are increasing year by year and have become a major public health problem that threatens human health. In recent years, despite tremendous efforts and significant progress in cancer treatment, treatment results are still limited. Among the three major cancer treatment methods (surgery, chemotherapy, and radiotherapy), chemotherapy plays a vital role in the treatment of various tumors. However, contemporary chemotherapy drugs are mainly administered intravenously, which has some undesirable limitations, including the uncontrollable distribution of drugs in tissues, resulting in cytotoxicity to normal cells, poor drug permeability, poor retention of passive diffusion, and frequent administration that can induce drug resistance, resulting in a narrow therapeutic window and poor drug safety. These factors are all caused by poor drug delivery in the body.

[0004] In order to address these obstacles, a variety of coping strategies have been developed. These include: (1) using nanocarriers to physically package drug molecules, improve drug properties, increase the solubility of poorly soluble drugs, and enhance drug enrichment in lesion tissues to improve drug bioavailability while reducing toxicity. However, nanomedicines packaged by physical interactions have poor in vivo stability and lack active targeting, which affects the delivery and release efficiency in the body. (2) Polymer drug conjugates: Polymer drug conjugates deliver drugs by coupling compounds to similar prodrugs, which can greatly improve the solubility of drugs, slowly release active drug ingredients, and have long-circulating pharmacokinetic characteristics. However, their inherent defects still exist, including cumbersome and complex synthesis procedures, difficulty in controlling drug loading, large molecular weight, poor membrane permeability, no active targeting, and potential systemic toxicity. These shortcomings further limit their clinical development.

[0005] Compared to traditional polymer prodrugs and nanocarriers, small molecule lipids (such as phospholipids, fatty acids, cholesterol, and glycerides) are abundant and offer advantages such as non-toxicity, good biocompatibility, and biodegradability, making them ideal for drug conjugate and nanocarrier design. This invention provides a novel class of phospholipid conjugates that are linked to drugs via responsive linkers and can self-assemble into nanoparticles. These phospholipid conjugates enhance the in vivo stability of nanodrugs and can release drug molecules in response to the lesion environment, thereby achieving excellent therapeutic effects.

[0006] Summary of the Invention

[0007] The technical problem to be solved by the present invention is to overcome the shortcomings of drug conjugates in the prior art. To this end, the present invention provides a phospholipid drug conjugate, a preparation method, and uses thereof. The phospholipid drug conjugate of the present invention has one or more of the following advantages: (1) excellent anti-tumor effect; (2) effective release of bioactive molecules / drugs; (3) nanometer size; and (4) good water solubility.

[0008] The present invention solves the technical problems of the present invention through the following technical solutions:

[0009] The present invention provides a nano-sized phospholipid drug conjugate containing a responsive linker as shown in formula (I).

[0010] or a stereoisomer of the phospholipid drug conjugate, a prodrug thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof,

[0011] in:

[0012] X 0 O or S;

[0013] X 1 O, S or NR 1a ;

[0014] X 2 O, S or NR 2a ;

[0015] L 1 , L 2 for non-existent or connected units;

[0016] M 0 OR 0a SR 0b NR 0c R 0d or T 0 -(LD) n ;

[0017] K 1 P 1 or Y 1 is the branching center, P 1 It is the pharmacokinetic regulation unit, M 1 T 1 -(LD) n ;

[0018] Y 2 It is a branching center;

[0019] M 2 Does not exist or T 2 -(LD) n ;

[0020] M 0 、M 1 and M 2 At least one of them contains an LD fragment;

[0021] Q 2a T 2a -R 1 Or not, Q 2b T 2b -R 2 Or not, Q 2a and Q 2b At least one exists;

[0022] T 0 、T 1 、T 2 Each independently selected from the absence or extension of a moiety;

[0023] T 2a 、T 2b for non-existent or connected units;

[0024] L is L 3 -L s , L 3 It is an environment-responsive connection unit, L s It is present or absent, and when present, it is a self-releasing connecting part;

[0025] D is a bioactive molecular fragment. When the conjugate contains multiple Ds, Ds can be the same or different.

[0026] R 1a and R 2a Each independently selected from hydrogen, C1-C 10 Alkyl, C1~C 10 Alkoxy, C3~C 10 Alkenyl, C3~C 10 Alkynyl, C3~C8 cycloalkyl, C2~C8 heterocycloalkyl, C6~C 10 Aryl, C5~C 10 heteroaryl or amino protecting groups;

[0027] R 0a Each independently selected from hydrogen, C1 to C 10 Alkyl, C1~C 10 Alkoxy, C3~C 10 Alkenyl, C3~C 10Alkynyl, C3~C8 cycloalkyl, C2~C8 heterocycloalkyl, C6~C 10 Aryl, C5~C 10 heteroaryl or hydroxy protecting groups;

[0028] R 0b Selected from hydrogen, C1~C 10 Alkyl, C1~C 10 Alkoxy, C3~C 10 Alkenyl, C3~C 10 Alkynyl, C3~C8 cycloalkyl, C2~C8 heterocycloalkyl, C6~C 10 Aryl, C5~C 10 heteroaryl or thiol protecting groups;

[0029] R 0c and R 0d Each independently selected from hydrogen, C1-C 10 Alkyl, C1~C 10 Alkoxy, C3~C 10 Alkenyl, C3~C 10 Alkynyl, C3~C8 cycloalkyl, C2~C8 heterocycloalkyl, C6~C 10 Aryl, C5~C 10 heteroaryl or amino protecting groups;

[0030] R 1 Selected from saturated or unsaturated C8~C 50 alkyl;

[0031] R 2 Selected from saturated or unsaturated C1~C 50 alkyl;

[0032] n is an integer from 1 to 10. When n>1, LD may be the same or different;

[0033] In the compound represented by formula (I), one or more hydrogen atoms attached to carbon atoms may be replaced by deuterium.

[0034] In the compound represented by formula (I), one or more hydrogen atoms connected to carbon atoms may be replaced by any substituent.

[0035] Phospholipid isomers include compounds with phosphorus as the chiral center, selected from:

[0036] or chiral mixtures thereof.

[0037] In some embodiments, R 1 and R 2 Each independently selected from saturated or unsaturated C1-C 50 Alkyl, cholesterol, vitamin A acid and tocopherol, the C1~C50 Alkyl, cholesterol, vitamin A acid and tocopherol are optionally substituted by one or more selected from hydroxyl, halogen, C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 Substituted by cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 Cycloalkyl and 3-12 membered heterocycloalkyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-6 The cycloalkyl group is substituted with a substituent.

[0038] In some embodiments, the C1 to C 50 The alkyl groups are each independently preferably C8 to C 50 Alkyl, more preferably C 10 ~C 30 Alkyl, more preferably C 10 ~C 20 alkyl.

[0039] In some embodiments, the present invention provides a nano-sized phospholipid drug conjugate containing a responsive linker or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate, characterized in that the phospholipid core in formula (I) One or more of the following conditions are met:

[0040] (1)

[0041] (2)

[0042] (3) where X 2 、X 1 、M 0 At least one atom connected to P is not an oxygen atom.

[0043] In some embodiments, the present invention provides a phospholipid drug conjugate having a nanometer size and containing a responsive linker, or a stereoisomer thereof, a prodrug, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, characterized in that the nanometer size is an average nanoparticle size in the range of 1-500 nanometers; preferably, the average particle size is in the range of 5-100 nanometers.

[0044] In some embodiments, the nanosized phospholipid drug conjugates containing responsive linkers provided by the present invention, or their stereoisomers, prodrugs, pharmaceutically acceptable salts, or pharmaceutically acceptable solvates, meet one or more of the following conditions:

[0045] (1)L 1 , L 2 is absent or present, when present, L 1 , L 2 Each is independently an alkane, a heterocycloalkane, an aromatic hydrocarbon, a heterocycloalkane, a substituted or unsubstituted amino acid or a polypeptide composed thereof, or a combination thereof, wherein L 1 , L 2 The functional group connecting parts at both ends are selected from:

[0046] Among them, R La 、R Lb 、R Lc 、R Ld Each independently selected from hydroxy, C1-C5 alkyl or C1-C5 alkoxy;

[0047] R Le is hydrogen, hydroxy, C1-C5 alkyl or C1-C5 alkoxy;

[0048] L 1a , L 1b Each independently selected from O, NR Lf , S, SS, Or directly connected (not present), R Lf is hydrogen or C1-C5 alkyl;

[0049] (2)T 0 、T 1 、T 2 Each is independent of the other. When present, T 0 、T 1 、T 2 Each is independently an alkane, a heterocycloalkane, an aromatic hydrocarbon, a heterocycloalkane, a substituted or unsubstituted amino acid or a polypeptide composed thereof, or a combination thereof, wherein T 0 、T 1 、T 2 The functional groups at the respective ends are selected from:

[0050] Among them, R La 、R Lb 、R Lc 、R Ld Each independently selected from hydroxy, C1-C5 alkyl or C1-C5 alkoxy;

[0051] R Le is hydrogen, hydroxy, C1-C5 alkyl or C1-C5 alkoxy;

[0052] L 1a , L 1b Each independently selected from O, NR Lf , S, SS, Or directly connected (not present), R Lf is hydrogen or C1-C5 alkyl;

[0053] (3)T 2a 、T 2b is absent or present, when present, T 2a 、T 2b Each is independently an alkane, a heterocycloalkane, an aromatic hydrocarbon, a heterocycloalkane, a substituted or unsubstituted amino acid or a polypeptide composed thereof, or a combination thereof, wherein T 2a 、T 2b The functional groups at the respective ends are selected from:

[0054] Among them, L 1a , L 1b Each independently selected from O, NR Lf , S, SS, Or directly connected (not present), R Lf is hydrogen or C1-C5 alkyl;

[0055] (4)L 3 selected from enzyme-responsive linking units, pH-sensitive linking units, reduction-responsive linking units, reactive oxygen species (ROS) stimulus-responsive linking units, or light-responsive linking units;

[0056] (5)Y 1 、Y 2 is an at least trifunctional branching center selected from trifunctional amino acids, polypeptides or the following structures:

[0057] Among them, R y0 selected from hydrogen, deuterium, halogen, nitro, cyano, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl, or a group containing a primary amine, secondary amine, tertiary amine, hydroxyl, sulfhydryl, carboxyl, ester, amide, boric acid, boric ester, phosphoric acid, sulfonic acid, sulfoxide, aldehyde, or ketone functional group;

[0058] Ry1 Selected from C1-C10 alkyl, C1-C10 alkoxy, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl;

[0059] R y2 selected from hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl, or a group containing a primary amine, secondary amine, tertiary amine, hydroxyl, sulfhydryl, carboxyl, ester, amide, boric acid, boric ester, phosphoric acid, sulfonic acid, sulfoxide, aldehyde, or ketone functional group;

[0060] A ring is C6~C 20 Aryl or C5~C 20 Heteroaryl; the C5~C 20 The heteroatom in the heteroaryl group is O, S or N, and the number of the heteroatoms is one or more. When there are multiple heteroatoms, the heteroatoms are the same or different;

[0061] Ring E is a C2-C8 heterocycloalkyl group; the heteroatom in the heterocycloalkyl group is O, S or N, and the number of the heteroatoms is one or more. When there are multiple heteroatoms, the heteroatoms are the same or different;

[0062] (6)P 1 The pharmacokinetic regulating unit is selected from polyethylene glycol, sugar, oligosaccharide and polysaccharide (such as hyaluronic acid, chondroitin sulfate, chitosan, heparin, tea polysaccharide, dextran, polysialic acid, chitosan, dextran, sodium alginate, cellulose, polysucrose, cyclodextrin), polysarcosine, polyphosphate, polyglycerol, polyvinylpyrrolidone, polyoxazoline, polyamino acid, polyacrylic acid residue, polymethacrylic acid residue, quaternary ammonium salt derivative, carboxyl derivative, sulfonic acid derivative, phosphate derivative, zwitterionic derivative or zwitterionic polymer.

[0063] In some embodiments, L 3 It is an environmentally responsive linker unit selected from enzyme-responsive linkers, pH-responsive linkers, light-responsive linkers and redox-responsive linkers, preferably selected from enzyme-responsive linkers, pH-responsive linkers and redox-responsive linkers, and more preferably an enzyme-responsive linker.

[0064] In some embodiments, the nanosized phospholipid drug conjugate containing a responsive linker provided by the present invention, or its stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate, is characterized in that it satisfies one or more of the following conditions:

[0065] (1) L 3It is an enzyme-responsive linker that can be cleaved by one or more of the following enzymes: secretory phospholipase A2, acid phosphatase, serum alkaline phosphatase, cytochrome P450, sulfatase, prostate-specific antigen, phospholipase A1, phospholipase A2, phospholipase B, phospholipase C, phospholipase D, neutrophil elastase, caspase-3, cathepsin, matrix metalloproteinase, β-glucuronidase, β-galactosidase, nitroreductase, reduced coenzyme II, aminopeptidase N, carboxylesterase, diaphorase, histone deacetylase, asparagine endopeptidase, urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor, collagenase; preferably cleaved by one or more of the following enzymes: caspase-3, cathepsin, matrix metalloproteinase, elastase or β-glucuronidase;

[0066] (2) L 3 A pH-responsive linker comprising one or more of a hydrazone, an imine, an oxime, a carboxylate, a thioester, a sulfate, a sulfonate, an orthoester, a carbonate, a carbamate, a substituted carbamate, a ketal, an acetal, a silyl ether, a phosphate, a borate, a phosphoramide, or a cis-aconitic acid group;

[0067] (3) L 3 It is a photoresponsive linker comprising the following structures: one or more of o-nitrobenzene, coumarin, benzoin, BODIPY or cyanine groups;

[0068] (4) L 3 It is a redox-responsive linker comprising one or more of the following structures: thioketal, phenylboronic acid, phenylboronic acid, oxalate, vinyl ether, thioether, aminoacrylate, disulfide, diselenyl, 2,4-dinitrobenzenesulfonate, 2-azidomethylbenzoate, 4-azidobenzyl, unsaturated acid ester or azobenzene group.

[0069] In some embodiments, the nanosized phospholipid drug conjugate containing a responsive linker provided by the present invention, or its stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate, is characterized in that it satisfies one or more of the following conditions:

[0070] (1)L 3It is an enzyme-responsive linker selected from a short peptide consisting of 2-10 amino acid residues Val, D-Val, Cit, Phe, Lys, Leu, Gly, Ala, Asn, Asp, Arg, Pro, Tyr, Ile, His, Ser, Gln, Glu, and Met, preferably a short peptide consisting of 2-10 amino acids selected from Val, Cit, Phe, Lys, Ala, Leu, Gly, Asn, and Asp; preferably, L 3 Selected from the following structures:

[0071] (2)L s When present, it is selected from the following structures:

[0072] Preferably, L s Selected from the following structures:

[0073] (3)L 3 is a pH-responsive linker selected from the following structures:

[0074] Among them, pm = 0 ~ 4; R p3 、R q1 is selected from H, substituted or unsubstituted C1-C10 alkyl;

[0075] Preferably, L 3 Selected from the following structures:

[0076] (4) L 3 is a light-responsive linker selected from the following structures:

[0077] (5)L 3 is a redox-responsive linker selected from the following structures:

[0078] In some embodiments, L 3 When it is a redox-responsive linker, it comprises the following structure:

[0079] In some embodiments, L 3 When it is a redox-responsive linker, it is selected from the following structures:

[0080] In some embodiments, the present invention provides a phospholipid drug conjugate having nanometer size and containing a responsive linker, or a stereoisomer, a prodrug, a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof, characterized in that: 1 Selected from:

[0081] (1) The polyethylene glycol derivative residue is selected from the following structures:

[0082] wherein pa1 is selected from an integer between 5 and 250, pr1 is selected from an integer between 0 and 8, and R p1 is hydrogen, C1-C10 alkyl, C1-C10 heteroalkyl, C3-C10 cycloalkyl, C3-C10 alkenyl, C3-C10 alkynyl or a hydroxy protecting group;

[0083] Preferably, pa1 is selected from an integer between 5 and 150, pr1 is selected from an integer between 0 and 8, and R p1 is hydrogen, a C1-C10 alkyl group or a hydroxyl protecting group;

[0084] More preferably, pa1 is selected from an integer between 10 and 60, pr1 is selected from an integer between 1 and 2, and R p1 is a C1-C10 alkyl group;

[0085] (2) The polycreatine derivative residue is selected from the following structures:

[0086] Wherein, pb1 is selected from an integer between 5 and 250, R p2 R is selected from hydrogen, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl or an amino protecting group; p3 Selected from OR p3a or NR p3b R p3c , where R p3a R is selected from hydrogen, C1-C10 alkyl, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl or C5-C10 heteroaryl; p3b 、R p3c Each is independently selected from hydrogen, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl, amino protecting group or R p3b and R p3c Together with the nitrogen atoms they are connected to, they form a C3-C10 ring structure;

[0087] Preferably, pb1 is selected from an integer between 5 and 150;

[0088] (3) The quaternary ammonium salt derivative residue is selected from the following structures:

[0089] Among them, pc1 is selected from integers between 0 and 10, A - Selected from R p3 、R p4 、R p5 Each is independently selected from C1-C10 alkyl, C1-C10 alkoxy, C3-C8 cycloalkyl or C2-C8 heterocycloalkyl;

[0090] (4) Zwitterionic derivatives or zwitterionic polymers:

[0091] in,

[0092] pd1 is selected from an integer between 5 and 250;

[0093] R p6 Selected from:

[0094] pc1, pe1 are integers between 0 and 10, A - Selected from R p3 、R p4 、R p5 Each is independently selected from C1-C10 alkyl, C1-C10 alkoxy, C3-C8 cycloalkyl or C2-C8 heterocycloalkyl; L p is absent or a connecting unit selected from alkanes, heterocycloalkanes, aromatic hydrocarbons, heterocycloalkanes or a combination thereof;

[0095] (5) Polysaccharide polymers:

[0096] Among them, pf1, pf2, pg1, pg2, ph1, ph2, pi1, and pi2 are selected from integers between 1 and 250.

[0097] In some embodiments, the polysaccharide polymer is selected from:

[0098] Among them, pf1, pf2, pg1, pg2, ph1, ph2, pi1, pi2, and m are each independently selected from an integer between 1 and 250, preferably an integer between 1 and 50, for example, an integer between 1 and 5, an integer between 4 and 24, or an integer between 5 and 25.

[0099] In some embodiments, pf1 is selected from an integer between 1-250, preferably an integer between 1-50, more preferably an integer between 1-20, such as an integer between 1-5.

[0100] In some embodiments, pf2 is selected from an integer between 1-250, preferably an integer between 1-50, such as an integer between 4-24.

[0101] In some embodiments, m is selected from an integer between 1-250, preferably an integer between 1-50, such as an integer between 5-25.

[0102] In some embodiments, the present invention provides a nanosized phospholipid drug conjugate containing a responsive linker, or a stereoisomer, prodrug, a pharmaceutically acceptable salt, or a pharmaceutically acceptable solvate thereof, characterized in that the bioactive molecule fragment D is selected from: small molecule drugs, peptides, proteins, nucleic acids, and carbohydrates, and specifically can be anesthetics, ophthalmic drugs, central nervous system drugs, pain therapeutics, antiarthritis drugs, anticonvulsants, antihistamines, antiulcer drugs, behavior correction drugs, respiratory drugs, antispasmodics, muscle relaxants, anti-inflammatory drugs, appetite suppressants, growth promoters, hemostatics, immunostimulants, immunomodulators, muscle relaxants, obesity therapeutics, osteoporosis drugs. The invention also includes but is not limited to: drugs, tranquilizers, tranquilizers, anti-migraine drugs, muscle contractants, anti-infectives, anti-rheumatic drugs, antimalarials, antiemetics, bronchodilators, antithrombotic drugs, antihypertensive drugs, cardiovascular drugs, antiarrhythmic drugs, antioxidants, antiasthmatic drugs, diuretics, lipid regulators, antiandrogens, antiparasitic drugs, anticoagulants, antitumor drugs, hypoglycemic drugs, nutritional agents, growth supplements, anti-enteritis drugs, vaccines, antibodies, radionuclides, diagnostic agents and contrast agents; preferably, they are selected from small molecule drugs or peptide drugs; more preferably, they are selected from antitumor drugs, such as cytotoxic drugs, small molecule targeted drugs, hormones, biological response modifiers (such as immunomodulators), tumor adjuvant drugs and radionuclide drugs.

[0103] In some embodiments, the nanosized phospholipid drug conjugate containing a responsive linker provided by the present invention, or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate, has a structure shown in formula (II):

[0104] Among them, X 0 、X 1 , L 1 、P 1 、X 2 , L 2 、Y 2 , Q 2a , Q 2b 、Y 2 , D is as defined in compound formula (I);

[0105] Where T 0 It exists or does not exist. When it exists, the connection point with the P end is selected from where R 01 is hydrogen, C1-C10 alkyl, C1-C10 alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

[0106] L is L 3 -L s , L 3 It is an environment-responsive connection unit, L s It exists or does not exist. When it exists, it is a self-releasing connecting part. When T 0 When L does not exist, 3 It is directly connected to the P end, and the functional group at the connection point is a nitrogen atom.

[0107] In some embodiments, the present invention provides a phospholipid drug conjugate having nanometer size and a responsive linker, or a stereoisomer, a prodrug, a pharmaceutically acceptable salt, or a pharmaceutically acceptable solvate thereof, characterized in that:

[0108] X 0 、X 1 、X 2 is O;

[0109] L 1 , L 2 Selected from:

[0110] For the P-terminal connection, P 1 or Y 2 End connection; l1a is selected from an integer between 0-10, preferably an integer selected from 1-3;

[0111] Y 2 Selected from

[0112] Preferably, selected from

[0113] More preferably, selected from For L 2 end connection;

[0114] P 1 Selected from the group consisting of: repeating unit number pa1, end group R p1 wherein pa1 is selected from an integer between 10 and 60, preferably an integer between 15 and 50, such as 21, 22, 44 or 45; and R p1 is a C1-C6 alkyl group, preferably a C1-C3 alkyl group, such as methyl, ethyl, n-propyl or isopropyl;

[0115] or P 1Selected from the repeating unit number pb1, the terminal group is R p2 Polycreatine derivative residues:

[0116] wherein pb1 is selected from an integer between 10 and 60, preferably an integer between 15 and 50; R p2 is selected from hydrogen, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl, or an amino protecting group;

[0117] Q 2a T 2a -R 1 , Q 2b T 2b -R 2 , T 2a 、T 2b The definition of is as described in compound formula (I), R 1 、R 2 Selected from:

[0118] wherein r1a and r1b are selected from integers of 1-30.

[0119] In some embodiments, X 0 It is O.

[0120] In some embodiments, X 1 It is O.

[0121] In some embodiments, X 2 It is O.

[0122] In some embodiments, M 0 -T 0 -(LD) n , preferably -LD, n is preferably 1.

[0123] In some embodiments, T 0 is absent or present, when present, T 0 Preferably selected from -NHCH2- and The C-terminus is preferably connected to L.

[0124] In some embodiments, when L is When T 0 for The C-terminus is connected to L

[0125] In some embodiments, M 2 In some embodiments, L 1 for Preferably With X 1 connect, With P 1 Connection, l1a is selected from an integer between 0-10, preferably selected from an integer between 0-5, such as 1 or 2.

[0126] In some embodiments, L 2 for With X 1 connect, With P 1 l1a is selected from an integer between 0-10, preferably an integer between 0-5, such as 0 or 2.

[0127] In some embodiments, K 1 P 1 , P 1 The pharmacokinetic regulating unit is selected from the group consisting of polyethylene glycol derivative residues, polycreatine derivative residues, quaternary ammonium salt derivative residues, zwitterion derivatives or zwitterion polymers, and polysaccharide polymers; preferably selected from the group consisting of polyethylene glycol derivative residues, polycreatine derivative residues, quaternary ammonium salt derivative residues and polysaccharide polymers; more preferably, the group consisting of polyethylene glycol derivative residues.

[0128] In some embodiments, the polyethylene glycol derivative residue is preferably

[0129] In some embodiments, pa1 is selected from integers between 5 and 250; preferably selected from integers between 5 and 150, such as 22, 45 or 113; more preferably selected from integers between 10 and 60, such as 45.

[0130] In some embodiments, pr1 is selected from an integer of 0-8, preferably an integer of 0-2, such as 0.

[0131] In some embodiments, R p1 It is hydrogen, C1-C10 alkyl, C1-C10 heteroalkyl, C3-C10 cycloalkyl, C3-C10 alkenyl, C3-C10 alkynyl or a hydroxyl-protecting group, preferably hydrogen, C1-C10 alkyl or a hydroxyl-protecting group, more preferably C1-C10 alkyl.

[0132] In some embodiments, the polycreatine derivative residue is preferably

[0133] In some embodiments, pb1 is selected from integers between 5 and 250, preferably selected from integers between 5 and 150, and more preferably selected from integers between 10 and 50, such as 28.

[0134] In some embodiments, R p2 Selected from hydrogen, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl and amino protecting group, preferably an amino protecting group, such as -C(O)CH3.

[0135] In some embodiments, the quaternary ammonium salt derivative residue is preferably

[0136] In some embodiments, R p3 、R p4 、R p5 Each is independently selected from C1-C10 alkyl, C1-C10 alkoxy, C3-C8 cycloalkyl or C2-C8 heterocycloalkyl, preferably C1-C10 alkyl, such as methyl.

[0137] In some embodiments, the polysaccharide polymer is preferably selected from:

[0138]

[0139] wherein pf1, pf2 and m are each independently selected from an integer between 1 and 250.

[0140] In some embodiments, Y 2 is a branching center, selected from Preferably selected from More preferred For L 2 End connection.

[0141] In some embodiments, T 2a 、T 2b is absent or present, when present, T 2a 、T 2b are each independently selected from -C(O)-.

[0142] In some embodiments, R 1 and R 2 Each independently selected from Preferably selected from wherein r1a and r1b are selected from integers of 1-30, Optionally, one or more selected from hydroxy, halogen, C1-10 Alkyl, C 1-10 Alkoxy, C 3-12 Substituted by cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 Cycloalkyl and 3-12 membered heterocycloalkyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-6 The cycloalkyl group is substituted with a substituent.

[0143] In some embodiments, R 1 and R 2 Each independently preferably selected from More preferably selected from Wherein, r1a and r1b are selected from integers of 1-30. Optionally, one or more selected from hydroxy, halogen, C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 Substituted by cycloalkyl and 3-12 membered heterocycloalkyl, the C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 Cycloalkyl and 3-12 membered heterocycloalkyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-6 The cycloalkyl group is substituted with a substituent.

[0144] In some embodiments, R 1 and R 2 Each independently selected from Preferably selected from In some embodiments, R 1 and R 2 Each independently preferably selected from More preferably

[0145] In some embodiments, Q 2a and Q 2b Each independently selected from Preferably selected from

[0146] In some embodiments, Q 2a and Q 2b Each independently preferably selected from More preferably selected from

[0147] In some embodiments, the present invention provides a nanosized phospholipid drug conjugate containing a responsive linker, or a stereoisomer, prodrug, a pharmaceutically acceptable salt, or a pharmaceutically acceptable solvate thereof, characterized in that the biologically active molecule fragment D is selected from:

[0148] (1) Cytotoxic drugs:

[0149] Camptothecins, a topoisomerase inhibitor represented by formula (III):

[0150] Among them, R x1 is selected from C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 cycloalkyl, C1-C4 deuterated alkyl, C2-C4 alkynyl or C2-C4 alkenyl;

[0151] R x2 、R x3 、R x4 and R x5 are each independently selected from a hydrogen atom, a deuterium atom, a halogen, a cyano group, a hydroxyl group, a thiol group, a sulfone group, a sulfoxide group, a substituted or unsubstituted amino group, a nitro group, an alkynyl group, an alkenyl group, an alkyl group, a haloalkyl group, a cycloalkyl group, an alkoxy group, an alkylthiol group, a heterocycloalkyl group, a deuterated alkyl group, an aryl group, or a heteroaryl group;

[0152] Or, R x1 and R x2 、R x2 and R x3 、R x3 and R x4 、R x4 and R x5 Each together with the carbon atom to which they are commonly attached forms a 5-7 membered cycloalkyl or heterocycloalkyl group;

[0153] X is selected from O or S;

[0154] Z is selected from OH or F;

[0155] n1 is selected from 0 or 1;

[0156] Preferably, it is selected from the following structures:

[0157] Tubulin inhibitors are preferably selected from:

[0158] Nucleic acid transcription, preferably, is selected from:

[0159] Drugs acting on DNA structures are preferably selected from:

[0160] wherein Rt1 is selected from hydrogen, alkyl, alkoxy, heteroalkyl, aryl or heteroaryl;

[0161] (2) Small molecule targeted drugs:

[0162] EGFR inhibitors, such as gefitinib, erlotinib, icotinib, afatinib, or osimertinib;

[0163] HER2 inhibitors, such as lapatinib, pyrotinib, neratinib, or tucatinib;

[0164] ALK inhibitors, such as crizotinib, alectinib, ceritinib, lorlatinib, or brigatinib;

[0165] MEK inhibitors, such as trametinib, cabozantinib, or bimetinib;

[0166] MET inhibitors, such as savotinib, savolitinib, or capmatinib;

[0167] BRAF inhibitors, such as sorafenib, dabrafenib, vemurafenib, vemurafenib, or contrafenib;

[0168] CDK4 / 6 inhibitors, such as palbociclib, ribociclib, abecilib, or

[0169] PARP inhibitors, such as olaparib, niraparib, rucaparib, or talazoparib;

[0170] BTK inhibitors, such as acalabrutinib, ibrutinib, zanubrutinib, tilarutib, or zanubrutinib;

[0171] Protease inhibitors, such as bortezomib, carfilzomib, or ixazomib;

[0172] mTOR inhibitors, such as temsirolimus or everolimus;

[0173] JAK inhibitors, such as ruxolitinib, tofacitinib, upadacitinib, baricitinib, dicolitinib, pilitinib, filgotinib, abrocitinib, paclitinib, or deuteroceletinib;

[0174] PI3K inhibitors, such as idelalisib, panicib, duvelisib, or upalisib;

[0175] VEGFR inhibitors, such as sorafenib, axitinib, apatinib, sunitinib, regorafenib, vandetanib, pazopanib, lenvatinib, cabozantinib, ponatinib, aflibercept, or fruquintinib;

[0176] PDGFR, c-Kit inhibitors, such as imatinib, nilotinib, or afatinib;

[0177] BCL inhibitors, such as venetoclax;

[0178] HDAC inhibitors, such as vorinostat, romidepsin, belinostat, panobinostat, or chidamide;

[0179] FLT3 inhibitors, such as midostaurin, gilteritinib, or quizartinib;

[0180] RET inhibitors, such as sepretib or pralsetinib;

[0181] KRAS inhibitors, such as sotolacizumab or adagracilib;

[0182] IDH inhibitors, such as emdesipine, ilotinib, or olutavirin;

[0183] BCR-ABL inhibitors, such as imatinib, dasatinib, nilotinib, ponatinib, or orebactinib;

[0184] (3) hormonal molecules, such as fulvestrant, abiraterone, prednisone, dexamethasone, bicalutamide, enzalutamide, tamoxifen, toremifene, letrozole, exemestane, goserelin, or leuprorelin;

[0185] (4) Immunotherapy molecules, such as small molecule PD-L1 inhibitors, TLR agonists, stimulator of interferon genes (STING) agonists, or indoleamine-2,3-dioxygenase (IDO) inhibitors;

[0186] (5) Other small molecules and peptides, such as lenalidomide, thalidomide, pomalidomide, ibedomide, octreotide, lanreotide or pasireotide.

[0187] In some embodiments, the drug acting on DNA structure is selected from:

[0188] wherein Rt1 is selected from hydrogen, alkyl, alkoxy, heteroalkyl, aryl or heteroaryl.

[0189] In some embodiments, the bioactive molecule fragment D is preferably selected from cytotoxic drugs, small molecule targeted drugs, hormone molecules, and other small molecules and polypeptides.

[0190] In some embodiments, the cytotoxic drug is preferably selected from the topoisomerase inhibitors camptothecins, tubulin inhibitors and drugs acting on DNA structures as shown in formula (III), more preferably selected from the topoisomerase inhibitors camptothecins and tubulin inhibitors as shown in formula (III);

[0191] The camptothecin class of topoisomerase inhibitors represented by formula (III) is preferably selected from:

[0192]

[0193] The tubulin inhibitors are preferably

[0194] The drug acting on DNA structure is preferably wherein Rt1 is selected from hydrogen, alkyl, alkoxy, heteroalkyl, aryl, and heteroaryl.

[0195] In some embodiments, the small molecule targeted drug is preferably selected from ALK inhibitors, CDK4 / 6 inhibitors and PARP inhibitors; the ALK inhibitor is preferably crizotinib; the CDK4 / 6 inhibitor is preferably selected from palbociclib and The PARP inhibitor is preferably niraparib.

[0196] In some embodiments, the hormone-like molecule is preferably fulvestrant.

[0197] In some embodiments, the other small molecules and polypeptides are preferably octreotide.

[0198] In some embodiments, D is selected from the following structures:

[0199] Preferably selected from

[0200] In some embodiments, L is selected from the following structures (which are connected to D by express):

[0201] Preferably selected from

[0202] In some embodiments, when L is When T 0 It is -NHCH2-, wherein the C-terminus is connected to L.

[0203] In some embodiments, when L is When T 0 for The C-terminus is connected to L.

[0204] In some embodiments, the present invention provides a nanosized phospholipid drug conjugate containing a responsive linker, or a stereoisomer, prodrug, a pharmaceutically acceptable salt, or a pharmaceutically acceptable solvate thereof, characterized in that LD is selected from any one of the following conditions:

[0205] (1) When the drug molecule D is of the following structure, the connection site with L is express:

[0206] L is selected from the following structures (the site where it is connected to the drug D is express):

[0207] (2) When the drug molecule D is of the following structure, the connection site with L is express:

[0208] L is selected from the following structures (the site where it is connected to the drug D is express):

[0209] In some embodiments, the nanosized phospholipid drug conjugate containing a responsive linker provided by the present invention, or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate, has a structure shown in formula (IV):

[0210] in,

[0211] (1) Selected from the following structures:

[0212] wherein x and q are selected from integers between 10 and 60; pf1, pf2, ph1, and ph2 are selected from integers between 1 and 100;

[0213] (2) Selected from the following structures:

[0214] wherein y, z, y1, y2, z1, z2, and z3 are selected from integers of 1 to 20;

[0215] (3) Selected from the following structures:

[0216] In some embodiments, Selected from the following structures:

[0217] In some embodiments, Selected from the following structures:

[0218] In some embodiments, Selected from the following structures:

[0219] In some embodiments, the nanosized phospholipid drug conjugate containing a responsive linker provided by the present invention, or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate, is selected from the following structures:

[0220] In some embodiments, the present invention provides a phospholipid drug conjugate having nanometer size and containing a responsive linker, or a stereoisomer, prodrug, a pharmaceutically acceptable salt, or a pharmaceutically acceptable solvate thereof, or a pharmaceutical composition thereof, for use in a drug for preventing, alleviating, and / or treating abnormal cell proliferation, infection (e.g., viral infection, such as HIV, HBV, etc.), inflammatory disease (e.g., rheumatoid arthritis, etc.), autoimmune disease (e.g., psoriasis, lupus erythematosus, diabetes, etc.), cardiovascular disease (e.g., stroke, myocardial infarction, atherosclerosis, etc.), or neurodegenerative disease (e.g., Alzheimer's disease, Parkinson's disease, etc.), wherein the abnormal cell proliferation disease can be cancer.

[0221] In some embodiments, the present invention provides a phospholipid drug conjugate having nanometer size and containing a responsive linker, or a stereoisomer, prodrug, a pharmaceutically acceptable salt, or a pharmaceutically acceptable solvate thereof, or a pharmaceutical composition thereof, for use in a drug for preventing, alleviating, and / or treating abnormal cell proliferation, infection (e.g., viral infection, such as HIV, HBV, etc.), inflammatory disease (e.g., rheumatoid arthritis, etc.), autoimmune disease (e.g., psoriasis, lupus erythematosus, diabetes, etc.), cardiovascular disease (e.g., stroke, myocardial infarction, atherosclerosis, etc.), eye disease, or neurodegenerative disease (e.g., Alzheimer's disease, Parkinson's disease, etc.), wherein the abnormal cell proliferation disease can be cancer.

[0222] In some embodiments, the present invention provides a phospholipid drug conjugate having a nanometer size and containing a responsive linker, or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate, or the pharmaceutical composition thereof, or its use in a drug for treating cancer, wherein the cancer includes breast cancer, ovarian cancer, brain cancer, prostate cancer, melanoma, nasopharyngeal cancer, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer (such as colon cancer, rectal cancer, etc.), lung cancer (such as small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma, undifferentiated cancer, etc.), kidney cancer, skin cancer, neuroblastoma, sarcoma, liposarcoma, osteochondrocyte sedimentation rate, atherosclerosis, leukemia, necrosis, necrosis factor alpha, leukemia ... tumors, bone cancer, osteosarcoma, seminoma, testicular tumors, uterine tumors (e.g., cervical cancer, endometrial cancer, etc.), head and neck tumors (e.g., laryngeal cancer, pharyngeal cancer, tongue cancer, etc.), multiple myeloma, malignant lymphomas (e.g., reticulum cell sarcoma, palatine lymphosarcoma, Hodgkin lymphoma, mantle cell lymphoma, etc.), polycythemia vera, leukemias (e.g., acute myeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, etc.), thyroid tumors, ureteral tumors, bladder tumors, gallbladder cancer, bile duct cancer, choriocarcinoma, or pediatric tumors (e.g., neuroblastoma, embryonic testicular cancer, retinoblastoma, etc.).

[0223] Unless otherwise stated, the terms used in this invention have the following meanings:

[0224] The amino protecting group, hydroxy protecting group and carboxyl protecting group of the present invention are suitable groups for protecting amino, hydroxy and carboxyl groups known in the art, as shown in the literature ("Protective Groups in Organic Synthesis", 5 Th Various types of protecting groups in Ed.TWGreene&P.GM Wuts).

[0225] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight or branched groups of 1 to 30 carbon atoms. Preferably, the alkyl group has 1 to 10 carbon atoms, more preferably 1 to 8 carbon atoms. Non-limiting examples include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3 The invention also includes but is not limited to: 1,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1,1,2-trimethylpropyl, 1-ethyl-2-methylpropyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 3,3-dimethylpentyl, 3,4-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, methyl, nonyl, decyl, undecyl, dodecyl, and various isomers thereof. The "alkyl" may be substituted or unsubstituted.

[0226] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic substituent comprising 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, and cyclooctyl. Non-limiting examples of polycyclic cycloalkyls include, but are not limited to, spirocyclic, fused, and bridged cycloalkyls. The cycloalkyl group may be substituted or unsubstituted.

[0227] The term "alkenyl" refers to an alkyl group as defined herein consisting of at least two carbon atoms and at least one carbon-carbon double bond, preferably a C2-C10 alkenyl group, more preferably a C2-C6 alkenyl group, such as ethenyl, propenyl, 1-propenyl, etc. The alkenyl group may be substituted or unsubstituted.

[0228] The term "alkynyl" refers to an alkyl group as defined herein consisting of at least two carbon atoms and at least one carbon-carbon triple bond, preferably a C2-C10 alkynyl group, more preferably a C2-C6 alkynyl group, such as ethynyl, 1-propynyl, 2-propynyl, etc. The "alkynyl" group may be substituted or unsubstituted.

[0229] The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent comprising 3 to 20 ring atoms, one or more of which is selected from N, O, Si, B, S(O) m 、P(O) m (wherein m is an integer from 0 to 2) heteroatoms, excluding the ring portion of -OO, -OS- or -SS-, and the remaining ring atoms are carbon. Preferably, the ring atoms are 3 to 12, of which 1 to 4 heteroatoms are contained. Non-limiting examples of monocyclic heterocycloalkyl groups include pyrrolyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, pyranyl, etc. Polycyclic heterocycloalkyl groups include spirocyclic, fused ring and bridged heterocycloalkyl groups. The "heterocycloalkyl" may be substituted or unsubstituted.

[0230] The term "alkoxy" refers to -O-(alkyl) and -O-(cycloalkyl), wherein alkyl and cycloalkyl are as defined herein. Non-limiting examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. The "alkoxy" may be substituted or unsubstituted.

[0231] The term "alkanethiol" refers to -S-(alkyl) and -S-(cycloalkyl), wherein alkyl and cycloalkyl are as defined herein. Non-limiting examples include, but are not limited to, methylmercapto, ethylmercapto, propylmercapto, butylmercapto, cyclopropylmercapto, cyclobutylmercapto, cyclopentylmercapto, cyclohexylmercapto, and the like. The "alkanethiol" may be substituted or unsubstituted.

[0232] The term "aryl" refers to any stable conjugated hydrocarbon ring system of 6 to 18 carbon atoms, preferably 6 to 10 carbon atoms, which may be a monocyclic, bicyclic, tricyclic, or higher-ring aromatic group, such as phenyl, naphthyl, and anthracene. The aryl group may include an aromatic ring with an aryl group on a heterocycloalkyl or cycloalkyl ring. The "aryl" group may be substituted or unsubstituted.

[0233] The term "heteroaryl" refers to an aromatic ring system in which at least one carbon atom in the ring is replaced by a heteroatom selected from N, O, or S. It is preferably a 5- to 7-membered monocyclic structure or a 7- to 12-membered bicyclic structure, and more preferably a 5- to 6-membered heteroaryl group, such as pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, thiazolyl, thienyl, pyrazinyl, triazolyl, tetrazolyl, oxazolyl, indazolyl, etc. The heteroaryl group may include a heteroaryl ring fused with a heteroaryl, heterocycloalkyl, or cycloalkyl ring. The "heteroaryl" group may be substituted or unsubstituted.

[0234] The term "hydroxy" refers to -OH.

[0235] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0236] The term "nitro" refers to -NO2.

[0237] The term "amino" refers to -NH2.

[0238] The term "cyano" refers to -CN.

[0239] The term "carboxylic acid" refers to -C(O)OH.

[0240] The term "mercapto" refers to -SH.

[0241] The term "substituted" refers to that one or more hydrogen atoms in a group are independently replaced by a corresponding number of deuterium or substituents.

[0242] The term "substituent" refers to a molecular group that replaces hydrogen in a compound, and may include, but is not limited to, halogen, alkyl, alkoxy, nitro, cyano, hydroxy, sulfone, sulfoxide, cycloalkyl, heterocycloalkyl, amino, substituted amino, azido, oxo, carboxyl, carboxylate, alkenyl, alkynyl, thiol, alkylthiol, aromatic, heterocyclic aromatic, or groups formed by combinations thereof.

[0243] The term "sulfone" refers to -S(O)2-alkyl, cycloalkyl, aryl or heteroaryl, wherein alkyl, cycloalkyl, aryl and heteroaryl are as defined above.

[0244] The term "sulfoxide" refers to -SO-alkyl, cycloalkyl, aryl or heteroaryl, wherein alkyl, cycloalkyl, aryl and heteroaryl are as defined above.

[0245] The term "carboxylate" refers to -C(O)O-alkyl, aryl or cycloalkyl, wherein alkyl, aryl and cycloalkyl are as defined above.

[0246] The term "amino acid" includes amino acids selected from naturally occurring α-L-amino acids, such as alanine (Ala or A), cysteine ​​(Cys or C), aspartic acid (Asp or D), glutamic acid (Glu or E), phenylalanine (Phe or F), glycine (Gly or G), histidine (His or H), isoleucine (Ile or I), lysine (Lys or K), leucine (Leu or L), methionine (Met or M), asparagine (Asn or N), proline (Pro or P), glutamine (Gln or Q), arginine (Arg or R), serine (Ser or S), threonine (Thr or T), valine (Val or V), tryptophan (Trp or W), tyrosine (Tyr or Y) and citrulline (Cit), as well as D-amino acids, synthetic α, β or γ amino acids, which can be substituted or unsubstituted.

[0247] The term "peptide" or "polypeptide" refers to natural and synthetic peptides and may comprise natural amino acids, unnatural amino acids, or a combination of natural and unnatural amino acids.

[0248] "Pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness of the free base without other toxic side effects. It can be an acidic group, a basic group or an amphoteric group. Non-limiting examples include, but are not limited to: acidic salts include hydrochlorides, hydrobromides, sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, nitrates, acetates, propionates, decanoates, octanoates, formates, acrylates, isobutyl phosphates, ... The pharmaceutically acceptable salts thereof include benzoate, methylbenzoate, phthalate, maleate, methanesulfonate, p-toluenesulfonate, benzenesulfonate, (D, L)-tartaric acid, citrate, maleate, (D, L-) malate, fumarate, stearate, oleate, cinnamate, laurate, glutamate, aspartate, trifluoromethanesulfonate, mandelate, ascorbate, salicylate, etc. When the compound of the present invention contains an acidic group, its pharmaceutically acceptable salt may also include: alkali metal salts (such as sodium salt or potassium salt), alkaline earth metal salts (such as calcium salt or magnesium salt), organic base salts (such as alkyl aromatic aminos, amino acids, etc.).

[0249] "Solvate" refers to an aggregate (or association) formed by one or more solvent molecules and the compound of the present invention. The solvents forming the solvate include, but are not limited to: water, dimethyl sulfoxide, methanol, ethanol, isopropanol, acetic acid, etc.

[0250] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically pharmaceutically acceptable salts or prodrugs, and other chemical components, as well as other components such as physiologically pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, promote absorption of the active ingredient, and thereby exert its biological activity.

[0251] "Prodrug" refers to a compound that can be converted into the biologically active compound of the present invention under physiological conditions (e.g., in vivo) or by solvolysis, and can be understood as a pharmaceutically acceptable metabolic precursor. A prodrug may be inactive or less active than the active parent compound, but can be rapidly converted in vivo to produce the parent compound of the present invention, thereby improving its solubility and certain metabolic properties in animals. Prodrugs include, for example, amino protecting groups, carboxyl protecting groups, and phospholipids.

[0252] "Isomers" refer to stereoisomers, including enantiomers and diastereomers. Cis-trans isomers are one type of diastereomers. The isomers in the compounds of the present invention may be enantiomers, diastereomers, and any mixture thereof, including free or salt-forming forms.

[0253] The term "effective amount" or "therapeutically effective amount" refers to a non-toxic amount of a drug or pharmaceutical agent sufficient to achieve the desired effect. The determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, as well as the specific active substance. The appropriate effective amount in each individual case can be determined by those skilled in the art through routine experimentation.

[0254] Unless otherwise indicated, the abbreviations of any protecting groups, amino acids and other compounds used in the present invention are based on their commonly used and recognized abbreviations or refer to IUPAC-IUBC Commission on Biochemical Nomenclature (see Biochem. 1972, 11, 942-944). BRIEF DESCRIPTION OF THE DRAWINGS

[0255] Figure 1 Nanoparticle diameter diagram of an embodiment of the present invention

[0256] FIG2 Evaluation of the efficacy of Example 1 of the present invention in a human liver cancer HepG2 xenograft mouse tumor model

[0257] FIG3 Evaluation of the efficacy of Examples 5, 7, 15 and 16 of the present invention in a human liver cancer HepG2 xenograft mouse tumor model

[0258] FIG4 Evaluation of the efficacy of Examples 1, 5 and 7 of the present invention in the human gastric cancer NCI-N87 xenograft mouse tumor model

[0259] Figure 5 Body weight changes of Examples 1, 5 and 7 of the present invention in the efficacy evaluation of human gastric cancer NCI-N87 xenograft mouse tumor model

[0260] FIG6 Evaluation of the efficacy of Example 5 and T-Dxd in a human colon cancer cell HT-29 xenograft mouse model

[0261] FIG7 Evaluation of the efficacy of Example 5 in a human fibrosarcoma cell HT-1080 xenograft mouse model

[0262] Figure 8 Pharmacodynamic study of Examples 5 and 46 in an orthotopic U87-MG glioma mouse model DETAILED DESCRIPTION

[0263] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0264] The abbreviations used in this invention are shown in the following table:

[0265] While the invention has been described with respect to specific embodiments thereof, certain modifications and equivalents will be apparent to one of ordinary skill in the art and are intended to be included within the scope of this invention.

[0266] HPLC analysis method for drug molecule content:

[0267] Instrument: Agilent 1260HPLC;

[0268] Chromatographic column: two-way valve, 1m damping tube, column temperature 40 degrees;

[0269] Phase A: methanol, flow rate 0.3 mL / min, detection wavelength 254 nm, isocratic: 0.0-5.0 min, 100%, phase A.

[0270] General HPLC analysis method for compound purity and content:

[0271] Chromatographic conditions:

[0272] Solvent gradient conditions:

[0273] The structures of all compounds of the present invention can be determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). - 6The unit of measurement is ppm. NMR measurements were performed on a Bruker AVANCE-400 spectrometer. The deuterated solvents tested were deuterated chloroform (CDCl3), deuterated methanol (MeOD), and deuterated dimethyl sulfoxide (DMSO-D6), with tetramethylsilane (TMS) as the internal standard.

[0274] Low-resolution mass spectra (MS) were measured on an Agilent 6120 quadruple LCMS mass spectrometer.

[0275] Example 1: Synthesis of Compound 1

[0276] Synthesis of compound 1-1

[0277] To a solution of D-alanine benzyl ester benzenesulfonate (10 g, 28.5 mmol, 1.2 eq) in acetonitrile (160 mL) were added N-Boc glycine (4.2 g, 23.7 mmol, 1.0 eq), DIPEA (3.67 g, 28.5 mmol, 1.2 eq), TCFH (7.99 g, 28.5 mmol, 1.2 eq), and N-methylimidazole (4.9 g, 60.0 mmol, 2.1 eq) at 0°C. The mixture was allowed to return to room temperature and stirred for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure at low temperature. The residue was purified on a silica gel column with PE / EA = 1:1 to afford the crude product. The crude product was further purified on a C18 column (CH3CN:H2O = 5-50%, 0.05% TFA) to afford 8.2 g of a pure white solid.

[0278] MS (ESI), m / z, 281.0 [M-55] + .

[0279] Synthesis of compound 1-2

[0280] To a methanol solution (200 mL) of 1-1 (8.2 g, 24.4 mmol, 1.0 eq) was added 10% Pd / C (800 mg) at 25°C. The mixture was replaced with H2 (balloon) and stirred at room temperature for 2 h under an H2 atmosphere. After completion of the reaction, MeOH (200 mL) was added, the mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure and dried in vacuo to yield 6.1 g of a pure white solid.

[0281] MS (ESI), m / z, 191.0 [M-55] + .

[0282] Synthesis of compounds 1-3

[0283] To a DMF solution of SN38 (3.0 g, 7.65 mmol, 1.0 eq) (120 mL) at 25°C was added TBDPSCl (7.4 g, 26.9 mmol, 3.5 eq) and then Et3N (3.6 g, 35.6 mmol, 4.5 eq) dropwise. The mixture was stirred at room temperature for 3 h. After completion of the reaction, the reaction solution was poured into ice water (400 mL), whereupon a large amount of solid precipitated. The mixture was filtered, the filter cake washed with water, and then pressed dry. The filter cake was dissolved in DCM (200 mL) and washed twice with saturated NaCl (150 mL x 2 to remove residual triethylamine acetate). The mixture was dried over anhydrous Na2SO4, concentrated under reduced pressure, and dried in vacuo to yield 4.3 g of a yellow solid.

[0284] MS (ESI), m / z, 631.20 [M+H] + .

[0285] Synthesis of compounds 1-4

[0286] To a CH2Cl2 solution (80 mL) of 1-3 (3.0 g, 4.76 mmol, 1.0 eq) at 25°C was added 1-2 (2.9 g, 11.9 mmol, 2.5 eq), DCC (2.5 g, 11.9 mmol, 2.5 eq), and DMAP (581 mg, 4.76 mmol, 1.0 eq) in that order. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was filtered, and the filtrate was concentrated under reduced pressure. Purification on a silica gel column (PE / EA = 1:1 as eluent) afforded 2.9 g of the pure product.

[0287] 1 H NMR(400MHz,Chloroform-d)δ8.02(d,J=9.2Hz,1H),7.76(ddd,J=8.0,3.1,1.4Hz,4H),7.50–7.35(m,7H),7 .12–7.04(m,2H),6.68(d,J=7.5Hz,1H),5.65(d,J=17.2Hz,1H),5.38(d,J=17.2Hz,1H),5.10(s,2H),4.79– 4.64(m,1H),3.94(d,J=15.6Hz,1H),3.71(dd,J=16.7,5.8Hz,1H),3.21(s,2H),2.64(q,J=7.6Hz,2H),2.38 –2.08(m,2H),1.52(d,J=8.0Hz,3H),1.19(s,9H),1.17(s,9H),0.96(t,J=7.5Hz,3H),0.89(t,J=8.1Hz,3H).

[0288] Synthesis of compounds 1-5

[0289] To a CH2Cl2 solution (6 mL) of compound 1-4 (2.2 g, 2.60 mmol, 1.0 eq) was added dropwise TFA (6 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated, and the residue was thoroughly concentrated using an oil pump. CH3CN (500 mL) was added and the mixture was further concentrated under reduced pressure to yield 2.1 g of a yellow solid.

[0290] MS (ESI), m / z, 759.20 [M+H] + .

[0291] Synthesis of compounds 1-6

[0292] At 25°C under N₂ protection, DMF (200 μL, 3.71 mmol, 5.0 eq) was added to a CH₂Cl₂ solution (20 mL) of DSPE-mPEG2000 (2.07 g, 0.742 mmol, 1.0 eq). The temperature was lowered to 0°C, and oxalyl chloride (600 μL, 6.68 mmol, 9.0 eq) was added dropwise to the reaction mixture. After the addition was complete, the mixture was stirred at room temperature for 3 h. After completion of the reaction, the mixture was concentrated under N₂ protection to afford 2.0 g of a crude white solid, which was used directly in the next step.

[0293] Synthesis of compounds 1-7

[0294] At room temperature (25°C) and under N2 protection, dissolve the crude product 1-6 (2.0 g, 0.713 mmol, 1.0 eq) in CH2Cl2 (15 mL). Cool to 0°C, and dropwise add a DCM solution (5 mL) of 1-5 (3.05 g, 3.56 mmol, 5.0 eq) and triethylamine (1.5 mL, 10.7 mmol, 15.0 eq) to the reaction solution. After the addition is complete, remove the ice-water bath and stir at room temperature for 2 h. After the reaction is complete, concentrate under reduced pressure, dissolve in DCM (50 mL), and wash three times with saturated NaCl (200 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was first purified by a normal phase column (first with PE / EA = 1:1 to 0:1, then with DCM / MeOH = 20:1 to 10:1 to 6:1), and then by a C18 reverse phase column (THF:H2O = 5-75%, t = 28 min, product peak gradient: 60%). The collected solution was concentrated under reduced pressure at low temperature (<30°C) to obtain 1.6 g of a light yellow pure product.

[0295] Synthesis of compound 1

[0296] To a THF solution of 1-7 (1.67 g, 0.49 mmol, 1.0 eq) (24 mL) at 0°C were added TBAF (980 μL, 0.98 mmol, 2.0 eq) and AcOH (143 mg, 2.45 mmol, 5.0 eq). The mixture was returned to room temperature and stirred for 2 h. The reaction mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed twice with saturated NaCl (150 mL x 2). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was first purified by silica gel column (DCM / MeOH = 9:1) and then by neutral alumina column to obtain compound 1, 770 mg, as a white solid.

[0297] MS (ESI), m / z, 606.9 [DAG-OH] + .693.3[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0298] 1 H NMR (400MHz, CDCl3, ppm): δ (ppm) 8.15-8.04 (m, 1H), 7.51-7.43 (m, 1H), 7.43-7.29 (m, 2H), 7.12 (s, 1H), 5.80-5.5 3(m,2H),5.46-5.34(m,1H),5.26-5.15(m,1H),4.81-4.69(m,1H),4.38-4.27(m,1H),4.26-3.98(m,8H),3.86-3.7 8(m,1H),3.75-3.53(m,203H),3.47-3.44(m,1H),3.43-3.35(m,5H),3.13-3.03(m,2H),2.36-2.25(m,6H),2.21-2 .10(m,2H),2.05-2.00(m,2H),1.64-1.52(m,8H),1.38-1.20(m,72H),0.99(t,J=8.0Hz,3H),0.88(t,J=8.0Hz,6H)

[0299] Example 2: Synthesis of Compound 2

[0300] Synthesis of compound 2-1

[0301] To a DMF solution of N-tert-butyloxycarbonyl-L-alanine (2.5 g, 13.2 mmol, 1.0 eq) in 25 mL of DMF was added cesium carbonate (8.6 g, 26.4 mmol, 2.0 eq) at 0°C. The mixture was stirred at room temperature for 30 min. Benzyl bromide (2.37 g, 13.86 mmol, 1.05 eq) was added dropwise. The mixture was stirred at room temperature for 16 h. After completion of the reaction, the reaction mixture was poured into water (150 mL), extracted with EA (100 mL x 2), washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by reverse phase column chromatography (CH3CN / H2O = 5-70%, 0.05% TFA) to yield 4.1 g of a pure white solid.

[0302] Synthesis of compound 2-2

[0303] At 25°C, 2-1 (4.1 g, 14.5 mmol, 1.0 eq) was dissolved in 4M HCl / dioxane (20 mL) and stirred at room temperature for 3 h. After the reaction was complete, the mixture was concentrated under reduced pressure at low temperature. The mixture was fully dried under vacuum to obtain 3.6 g of pure white solid.

[0304] MS (ESI), m / z, 179.80 [M+H] + .

[0305] Synthesis of compound 2-3

[0306] To a solution of 2-2 (3.18 g, 14.76 mmol, 1.2 eq) in acetonitrile (50 mL) at 0°C were added N-Boc glycine (2.15 g, 12.30 mmol, 1.0 eq), DIPEA (1.9 g, 14.76 mmol, 1.2 eq), TCFH (4.14 g, 14.76 mmol, 1.2 eq), and N-methylimidazole (3.54 g, 43.1 mmol, 3.5 eq). The mixture was allowed to return to room temperature and stirred for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure at low temperature and purified on a silica gel column (PE / EA = 2:1 to 1:1) to afford 3.5 g of a pure white solid.

[0307] MS (ESI), m / z, 280.80 [M-55] + .

[0308] Synthesis of Compound 2-4

[0309] To a solution of 2-3 (3.5 g, 10.4 mmol, 1.0 eq) in methanol (100 mL) was added 10% Pd / C (350 mg) at 25°C. The mixture was replaced with H2 (balloon) and stirred at room temperature for 4 h under an H2 atmosphere. After completion of the reaction, MeOH (200 mL) was added, the mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure and purified on a reverse phase column (CH3CN:H2O = 5-60%, 0.05% TFA) to afford a pure white solid (2.85 g).

[0310] MS (ESI), m / z, 190.80 [M-55] + .

[0311] Synthesis of Compound 2-5

[0312] To a CH2Cl2 solution (20 mL) of 1-3 (2.66 g, 4.22 mmol, 1.0 eq) at 25°C were added 2-4 (2.603 g, 10.54 mmol, 2.5 eq), EDCI (2.42 g, 12.67 mmol, 3.0 eq), and DMAP (1.03 g, 8.44 mmol, 2.0 eq) in that order. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure and purified on a silica gel column (PE / EA = 2:1 to 1:1 as eluent) to obtain 2.25 g of the pure product.

[0313] Synthesis of Compound 2-6

[0314] To a CH2Cl2 solution (20 mL) of 2-5 (2.4 g, 2.79 mmol, 1.0 eq) was added dropwise TFA (20 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the product as a yellow solid (2.6 g).

[0315] MS (ESI), m / z, 759.20 [M-55] + .

[0316] Synthesis of Compound 2-7

[0317] At room temperature (25°C) under N₂ protection, dissolve the crude product 1-6 (800 mg, 0.285 mmol, 1.0 eq) in CH₂Cl₂ (20 mL). Cool to 0°C, and dropwise add a solution of 2-6 (1.22 g, 1.425 mmol, 5.0 eq) in DCM (5 mL) and triethylamine (288 mg, 2.85 mmol, 10 eq) to the reaction mixture. After the additions were complete, remove the ice-water bath and stir at room temperature for 1 h. After completion of the reaction, concentrate under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (150 mL x 3). Combine the organic phases, dry over anhydrous Na₂SO₄, and concentrate under reduced pressure. The crude product was purified by normal phase column chromatography (EA / MeOH = 1:0 to 9:1, then switched to DCM / MeOH = 95:5 to 90:10) to yield 745 mg of crude product. The crude product was then purified by reverse phase column (THF / H2O=50:50), and the obtained crude product was further purified by neutral alumina column to obtain 210 mg of pure product.

[0318] Synthesis of compound 2

[0319] To a THF solution (15 mL) of 2-7 (200 mg, 0.057 mmol, 1.0 eq) was added a pyridine solution of hydrogen fluoride (70%, 80 mg, 0.57 mmol, 10 eq) at 0°C. The mixture was stirred at room temperature for 2 h. A small amount of saturated aqueous NaHCO₃ was added to neutralize the pH. The mixture was concentrated under reduced pressure, and the crude product was purified on a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 110 mg of the pure product.

[0320] MS (ESI), m / z, 607.0 [DAG-OH] + .693.5[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0321] Example 3: Synthesis of Compound 3

[0322] Synthesis of compound 3-1

[0323] To a DMF solution (20 mL) of N-Boc glycine (3.3 g, 22.5 mmol, 1.1 eq) at 0°C were added L-valine benzyl ester (4.2 g, 23.7 mmol, 1.0 eq) and HATU (9.6 g, 31.3 mmol, 1.5 eq). DIPEA (4.4 g, 41.0 mmol, 2.0 eq) was slowly added dropwise. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction solution was poured into water (150 mL) and extracted with EA (100 mL x 2). The mixture was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified on a silica gel column (PE / EA = 1:1) to obtain 4.7 g of a pure white solid.

[0324] MS (ESI), m / z, 364.80 [M+H] + .

[0325] Synthesis of compound 3-2

[0326] To a methanol solution (300 mL) of 3-1 (6.1 g, 15.8 mmol, 1.0 eq) was added 10% Pd / C (600 mg) at 25°C. The mixture was replaced with H2 (balloon) and stirred at room temperature for 4 h. After completion of the reaction, MeOH (200 mL) was added, the mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure and purified on a reverse phase column (CH3CN:H2O = 5-60%) to afford 2.9 g of a pure white solid.

[0327] MS (ESI), m / z, 218.80 [M-55] + .

[0328] Synthesis of compound 3-3

[0329] To a CH2Cl2 solution (25 mL) of 1-3 (1.7 g, 2.68 mmol, 1.0 eq) was added 3-2 (2.2 g, 8.03 mmol, 3.0 eq), DCC (1.7 g, 8.03 mmol, 3.0 eq), and DMAP (327 mg, 2.68 mmol, 1.0 eq) in that order at 25°C. The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The mixture was purified on a silica gel column (PE / EA = 1:1 as eluent) to afford 2.6 g of a yellow solid.

[0330] MS (ESI), m / z, 886.60 [M+H] + .

[0331] Synthesis of compound 3-4

[0332] To a CH2Cl2 solution (15 mL) of 3-3 (2.6 g, 2.93 mmol, 1.0 eq) was added dropwise TFA (15 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain the product as a yellow solid (2.7 g).

[0333] MS (ESI), m / z, 786.60 [M+H] + .

[0334] Synthesis of compound 3-5

[0335] At room temperature (25°C) under N2 protection, dissolve the crude product 1-6 (800 mg, 0.285 mmol, 1.0 eq) in CH2Cl2 (20 mL). Cool to 0°C, and dropwise add a solution of 3-4 (1.26 g, 1.425 mmol, 5.0 eq) in DCM (5 mL) and triethylamine (288 mg, 2.85 mmol, 10 eq) to the reaction mixture. After the addition is complete, remove the ice-water bath and stir at room temperature for 1 h. After completion of the reaction, concentrate under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (150 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was passed through a normal phase column, first using an EA / MeOH = 1:0 to 10:1 elution system, and then switched to an elution system DCM / MeOH = 10:1 to 5:1 to obtain 589 mg of crude product. The crude product was further purified by a neutral alumina column (DCM / MeOH = 98:2 to 97:3) to obtain 277 mg of a light yellow solid.

[0336] Synthesis of compound 3

[0337] To a THF solution (10 mL) of 3-5 (200 mg, 0.056 mmol, 1.0 eq) was added a pyridine solution of hydrogen fluoride (70%, 100 mg, 0.707 mmol, 12.6 eq) at 0°C. The mixture was stirred at room temperature for 2 h. A small amount of saturated aqueous NaHCO₃ was added to neutralize the pH. The mixture was concentrated under reduced pressure and the crude product was passed through a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 130 mg of the pure product.

[0338] MS (ESI), m / z, 607.1 [DAG-OH] + .700.6[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0339] Example 4: Synthesis of Compound 4

[0340] Synthesis of compound 4-1

[0341] To a solution of D-valine benzyl ester (3.21 g, 13.2 mmol, 1.2 eq) in acetonitrile (50 mL) at 0°C were added N-Boc glycine (1.93 g, 11.0 mmol, 1.0 eq), DIPEA (1.7 g, 13.2 mmol, 1.2 eq), TCFH (3.7 g, 13.2 mmol, 1.2 eq), and N-methylimidazole (3.16 g, 38.5 mmol, 3.5 eq). The mixture was allowed to cool to room temperature and stirred for 2 h. After completion of the reaction, the acetonitrile was concentrated under reduced pressure at low temperature. The residue was purified on a silica gel column (PE / EA = 1:1) to afford the crude product. The crude product was further purified on a reverse phase C18 column (CH3CN:H2O = 5-50%) to afford 3.5 g of a pure white solid.

[0342] MS (ESI), m / z, 308.80 [M-55] + .

[0343] Synthesis of compound 4-2

[0344] To a methanol solution (100 mL) of 4-1 (2.6 g, 7.13 mmol, 1.0 eq) was added 10% Pd / C (260 mg) at room temperature (25°C). H₂ replacement (balloon) was performed and the mixture was stirred at room temperature for 4 h. After completion of the reaction, MeOH (200 mL) was added and the mixture was filtered through Celite. The filtrate was concentrated under reduced pressure and purified on a reverse phase column (CH₃CN:H₂O = 5-60%) to afford 2.6 g of a pure white solid.

[0345] MS (ESI), m / z, 218.80 [M-55] + .

[0346] Synthesis of compound 4-3

[0347] To a CH2Cl2 solution (20 mL) of 1-3 (1.9 g, 2.94 mmol, 1.0 eq) at 25°C were added 4-2 (2.01 g, 7.34 mmol, 2.5 eq), EDCI (1.7 g, 8.82 mmol, 3.0 eq), and DMAP (717 mg, 5.88 mmol, 2.0 eq) in that order. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure and purified on a silica gel column (PE / EA = 2:1 to 1:1 as eluent) to obtain 2.25 g of the pure product.

[0348] MS (ESI), m / z, 886.60 [M+H] + .

[0349] Synthesis of compound 4-4

[0350] To a CH2Cl2 solution (15 mL) of 4-3 (2.2 g, 2.60 mmol, 1.0 eq) was added dropwise TFA (15 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the product as a yellow solid (2.2 g).

[0351] MS (ESI), m / z, 786.60 [M+H] + .

[0352] Synthesis of compound 4-5

[0353] At room temperature (25°C) under N₂ protection, dissolve the crude product 1-6 (800 mg, 0.285 mmol, 1.0 eq) in CH₂Cl₂ (20 mL). Cool to 0°C, and dropwise add a solution of 4-4 (1.26 g, 1.425 mmol, 5.0 eq) in DCM (5 mL) and triethylamine (288 mg, 2.85 mmol, 10 eq) to the reaction mixture. After the additions are complete, remove the ice-water bath and stir at room temperature for 1 h. After completion of the reaction, concentrate under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (150 mL x 3). Combine the organic phases, dry over anhydrous Na₂SO₄, and concentrate under reduced pressure. The crude product is then passed through a normal phase column, initially using an EA / MeOH = 1:0 to 9:1 elution system, then switching to a DCM / MeOH = 95:5 to 90:10 elution system to yield 745 mg of crude product. The crude product was purified by reverse phase column (THF / H2O=50:50) and finally purified by neutral alumina column to obtain 280 mg of pure product.

[0354] Synthesis of compound 4

[0355] To a THF solution (15 mL) of 4-5 (210 mg, 0.059 mmol, 1.0 eq) was added a pyridine solution of hydrogen fluoride (70%, 0.59 mmol, 10 eq) at 0°C. The mixture was stirred at room temperature for 2 h. A small amount of saturated sodium bicarbonate solution was added to neutralize the pH. The mixture was concentrated under reduced pressure, and the crude product was purified on a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 120 mg of the pure product.

[0356] MS (ESI), m / z, 606.7 [DAG-OH] + .700.8[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0357] Example 5: Synthesis of Compound 5

[0358] Synthesis of compound 5-1

[0359] To a solution of glycine benzyl ester hydrochloride (6.7 g, 31.7 mmol, 1.2 eq) in acetonitrile (80 mL) at 0°C were added N-tert-butyloxycarbonyl-L-alanine (5.0 g, 26.4 mmol, 1.0 eq), DIPEA (4.1 g, 31.7 mmol, 1.2 eq), TCFH (8.9 g, 31.7 mmol, 1.2 eq), and N-methylimidazole (7.6 g, 92.4 mmol, 3.5 eq). After addition, the mixture was allowed to return to room temperature and stirred for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure at low temperature. The residue was purified on a silica gel column (PE / EA = 1:1) to yield 6.4 g of a white solid.

[0360] MS (ESI), m / z, 280.80 [M-55] + .

[0361] Synthesis of compound 5-2

[0362] To a solution of 5-1 (7.0 g, 20.8 mmol, 1.0 eq) in methanol (200 mL) was added 10% Pd / C (700 mg) at 25°C. H₂ replacement (balloon) was performed and the mixture was stirred at room temperature under H₂ for 2 h. After completion of the reaction, MeOH (200 mL) was added and the mixture was filtered through Celite. The filtrate was concentrated under reduced pressure and dried in vacuo to yield 4.1 g of a white solid.

[0363] MS (ESI), m / z, 190.80 [M-55] + .

[0364] Synthesis of compound 5-3

[0365] To a CH2Cl2 solution (70 mL) of 1-3 (2.1 g, 3.3 mmol, 1.0 eq) was added 5-2 (2.01 g, 8.16 mmol, 2.5 eq), EDCI (1.9 g, 9.9 mmol, 3.0 eq), and DMAP (610 mg, 5.0 mmol, 1.5 eq) in sequence at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure and purified on a silica gel column (PE / EA = 2:1 to 1:1 as eluent) to give 2.2 g of a yellow-green solid.

[0366] MS (ESI), m / z, 859.80 [M+H] + .

[0367] Synthesis of compound 5-4

[0368] To a CH2Cl2 solution (15 mL) of 5-3 (2.5 g, 2.91 mmol, 1.0 eq) was added dropwise TFA (15 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the product as a yellow solid (2.6 g).

[0369] MS (ESI), m / z, 759.80 [M+H] + .

[0370] Synthesis of compound 5-5

[0371] At 25°C under N2 protection, crude 1-6 (800 mg, 0.285 mmol, 1.0 eq) was dissolved in CH2Cl2 (20 mL). The temperature was lowered to 0°C, and a DCM solution (5 mL) of 5-4 (1.22 g, 1.425 mmol, 5.0 eq) and triethylamine (288 mg, 2.85 mmol, 10 eq) were added dropwise to the reaction mixture. After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed three times with water (150 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was then passed through a normal phase column, initially using an EA / MeOH = 1:0-10:1 elution system, then switching to a DCM / MeOH = 15:1-10:1 elution system to obtain 290 mg of the product.

[0372] Synthesis of compound 5

[0373] To a THF solution of 5-5 (440 mg, 0.125 mmol, 1.0 eq) (15 mL) at 0°C were added TBAF (154 μL, 0.154 mmol, 1.23 eq) and acetic acid (38 mg, 0.633 mmol, 5.0 eq). The mixture was returned to room temperature and stirred for 2 h. The reaction mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed three times with water (150 mL x 2). The combined organic phases were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude product was purified on a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 170 mg of the pure product.

[0374] MS (ESI), m / z, 607.0 [DAG-OH] + .693.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0375] 1H NMR(400MHz,Chloroform-d)δ(ppm)8.63(s,1H),8.06(d,J=9.2Hz,1H),7.58–7.33(m,3H),7.21(d,J=2.6Hz,1H), 7.10(ddd,J=18.3,8.5,2.6Hz,1H),5.69(dt,J=17.0,2.6Hz,1H),5.57(d,J=16.1Hz,1H),5.37(d,J=17.1Hz,1H),5 .26–5.05(m,3H),4.40(dd,J=17.9,6.0Hz,1H),4.30–3.74(m,6H),3.71-3.52(s,193H),3.38(s,3H),3.31–3.00(m ,2H),2.38–2.17(m,5H),1.57(q,J=7.1Hz,5H),1.46–1.14(m,116H),1.00(t,J=7.4Hz,4H),0.88(t,J=6.7Hz,9H).

[0376] Example 6: Synthesis of Compound 6

[0377] Synthesis of compound 6-1

[0378] To a solution of glycine benzyl ester hydrochloride (6.7 g, 31.7 mmol, 1.2 eq) in acetonitrile (80 mL) at 0°C were added N-tert-butyloxycarbonyl-D-alanine (5.0 g, 26.4 mmol, 1.0 eq), DIPEA (4.1 g, 31.7 mmol, 1.2 eq), TCFH (8.9 g, 31.7 mmol, 1.2 eq), and N-methylimidazole (7.6 g, 92.4 mmol, 3.5 eq). The mixture was allowed to cool to room temperature and stirred for 2 h. After completion of the reaction, the acetonitrile was concentrated under reduced pressure at low temperature. The residue was purified on a silica gel column (PE / EA = 1:1) to yield 6.4 g of a white solid.

[0379] MS (ESI), m / z, 280.80 [M-55] + .

[0380] Synthesis of compound 6-2

[0381] To a solution of 6-1 (7.0 g, 20.8 mmol, 1.0 eq) in methanol (200 mL) was added 10% Pd / C (700 mg) at 25°C. H₂ replacement (balloon) was performed and the mixture was stirred at room temperature under H₂ for 2 h. After completion of the reaction, MeOH (200 mL) was added and the mixture was filtered through Celite. The filtrate was concentrated under reduced pressure and dried in vacuo to yield 4.1 g of a white solid.

[0382] MS (ESI), m / z, 190.80 [M-55] + .

[0383] Synthesis of compound 6-3

[0384] To a CH2Cl2 solution (70 mL) of 1-3 (2.1 g, 3.3 mmol, 1.0 eq) was added 6-2 (2.01 g, 8.16 mmol, 2.5 eq), EDCI (1.9 g, 9.9 mmol, 3.0 eq), and DMAP (610 mg, 5.0 mmol, 1.5 eq) in sequence at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure and purified on a silica gel column (PE / EA = 2:1 to 1:1 as eluent) to give 2.1 g of a yellow-green solid.

[0385] MS (ESI), m / z, 859.80 [M+H] + .

[0386] Synthesis of compound 6-4

[0387] To a CH2Cl2 solution (15 mL) of 6-3 (2.5 g, 2.91 mmol, 1.0 eq) was added dropwise TFA (15 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the product as a yellow solid (2.9 g).

[0388] MS (ESI), m / z, 759.80 [M+H] + .

[0389] Synthesis of compound 6-5

[0390] At room temperature (25°C) under N₂ protection, dissolve the crude product 1-6 (800 mg, 0.285 mmol, 1.0 eq) in CH₂Cl₂ (20 mL). Cool to 0°C, and dropwise add a DCM solution (5 mL) of 6-4 (1.22 g, 1.425 mmol, 5.0 eq) and triethylamine (288 mg, 2.85 mmol, 10 eq) to the reaction mixture. After the additions are complete, remove the ice-water bath and stir at room temperature for 1 h. After completion of the reaction, concentrate under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (150 mL x 3). Combine the organic phases, dry over anhydrous Na₂SO₄, and concentrate under reduced pressure. The crude product is then passed through a normal phase column, initially using an EA / MeOH = 1:0-10:1 elution system, then switching to a DCM / MeOH = 15:1-10:1 elution system to yield 170 mg of product.

[0391] Synthesis of compound 6

[0392] To a THF solution of 6-5 (170 mg, 0.048 mmol, 1.0 eq) (15 mL) at 0°C was added TBAF (60 μL, 0.060 mmol, 1.25 eq) and acetic acid (15 mg, 0.25 mmol, 5.2 eq). The mixture was returned to room temperature and stirred for 2 h. The reaction mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed three times with water (150 mL x 2). The combined organic phases were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude product was purified on a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 95 mg of the pure product.

[0393] MS (ESI), m / z, 606.9 [DAG-OH] + .693.3[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0394] Example 7: Synthesis of Compound 7

[0395] Synthesis of compound 7-1

[0396] To a DMF solution of glycine benzyl ester hydrochloride (4.2 g, 20 mmol, 1.0 eq) in 20 mL at 0°C was added BOC-glycine (3.5 g, 20 mmol, 1.0 eq) and HATU (11.4 g, 30 mmol, 1.5 eq). DIPEA (7.74 g, 60 mmol, 3.0 eq) was slowly added dropwise. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction solution was poured into water (150 mL) and extracted with EA (100 mL x 2). The mixture was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified on a silica gel column (PE / EA = 1:1) to yield 4.8 g of a pure white solid.

[0397] MS (ESI), m / z, 266.80 [M+H] + .

[0398] Synthesis of compound 7-2

[0399] To a methanol solution (100 mL) of 7-1 (5.0 g, 15.5 mmol, 1.0 eq) was added 10% Pd / C (500 mg) at 25°C. The mixture was replaced with H2 (balloon) and stirred at room temperature for 2 h. After completion of the reaction, MeOH (200 mL) was added, the mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure. The filtrate was purified by reverse phase column chromatography (CH3CN:H2O = 5-70%) to afford 3.0 g of a white solid.

[0400] MS (ESI), m / z, 176.80 [M-55] + .

[0401] Synthesis of compound 7-3

[0402] To a CH2Cl2 solution (20 mL) of 1-3 (3.4 g, 5.39 mmol, 1.0 eq) was added 7-2 (2.5 g, 10.78 mmol, 2.0 eq), EDCI (3.1 g, 16.17 mmol, 3.0 eq), and DMAP (1.32 g, 10.78 mmol, 2.0 eq) in sequence at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure and purified on a silica gel column (PE / EA = 2:1 to 1:1 as eluent) to afford 2.0 g of a yellow-green solid.

[0403] MS (ESI), m / z, 844.60 [M+H] + .

[0404] Synthesis of compound 7-4

[0405] To a solution of 7-3 (2.0 g, 2.36 mmol, 1.0 eq) in CH2Cl2 (10 mL) was added dropwise TFA (10 mL) at 25°C. The mixture was stirred at room temperature for 2 h. Upon completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was pumped dry using an oil pump. Acetonitrile (500 mL x 3) was added and the mixture was concentrated under reduced pressure to remove the residual TFA, yielding 2.0 g of a yellow solid.

[0406] MS (ESI), m / z, 744.60 [M+H] + .

[0407] Synthesis of compound 7-5

[0408] At room temperature (25°C) under N₂ protection, CH₂Cl₂ (20 mL) was added to the crude product 1-6 (800 mg, 0.29 mmol, 1.0 eq) to dissolve it. The temperature was lowered to 0°C, and a DCM solution (5 mL) of 7-4 (1.2 g, 1.43 mmol, 5.0 eq) and triethylamine (288 mg, 2.85 mmol, 10 eq) were added dropwise to the reaction mixture. After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed three times with water (150 mL x 3). The combined organic phases were dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The crude product was then purified on a normal phase column using an EA / MeOH = 1:0-10:1 elution system, then switched to a DCM / MeOH = 15:1-10:1 elution system to obtain the crude product. The crude product was finally purified on a neutral alumina column to obtain 290 mg of pure product.

[0409] Synthesis of compound 7

[0410] To a THF solution (3 mL) of 7-5 (140 mg, 0.042 mmol, 1.0 eq) was added a pyridine solution of hydrogen fluoride (70%, 60 mg, 0.42 mmol, 10 eq) at 0°C. The mixture was stirred at room temperature for 2 h. A small amount of saturated aqueous NaHCO₃ was added to neutralize the pH. The mixture was concentrated under reduced pressure, and the crude product was passed through a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 90 mg of the pure product.

[0411] MS (ESI), m / z, 606.8 [DAG-OH] + .689.8[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0412] 1 H NMR(400MHz,Chloroform-d)δ8.50(s,1H),8.12–8.00(m,1H),7.93(d,J=8.4Hz,1H),7.55–7.32(m,2H) ,7.18(d,J=1.9Hz,1H),7.10(d,J=8.6Hz,1H),5.76–5.53(m,2H),5.37(d,J=17.0Hz,1H),5.26–5.04(m ,3H),4.48–3.75(m,12H),3.72–3.41(m,165H),3.37(s,3H),3.19(dq,J=86.4,7.7,6.5Hz,3H),2.39–2 .09(m,6H),1.58(q,J=9.2,8.4Hz,5H),1.45–1.15(m,91H),0.99(t,J=7.4Hz,3H),0.91–0.82(m,14H).

[0413] Example 8: Synthesis of Compound 8

[0414] Synthesis of compound 8-1

[0415] To a solution of D-alanine benzyl ester benzenesulfonate (10.2 g, 29.07 mmol, 1.1 eq) in acetonitrile (80 mL) at 0°C were added N-Boc-β-alanine (5.0 g, 26.42 mmol, 1.0 eq), DIPEA (4.09 g, 31.7 mmol, 1.2 eq), TCFH (8.9 g, 31.7 mmol, 1.2 eq), and N-methylimidazole (8.3 g, 101.75 mmol, 3.5 eq). The mixture was allowed to cool to room temperature and stirred for 2 h. After completion of the reaction, the acetonitrile was concentrated under reduced pressure at low temperature. The residue was purified on a silica gel column (PE / EA = 1:1) to afford 8.8 g of a white solid.

[0416] MS (ESI), m / z, 295.20 [M-55] + .

[0417] Synthesis of compound 8-2

[0418] To a solution of 8-1 (8.8 g, 25.11 mmol, 1.0 eq) in methanol (100 mL) was added 10% Pd / C (880 mg) at 25°C. H₂ replacement (balloon) was performed and the mixture was stirred at room temperature under H₂ for 2 h. After completion of the reaction, MeOH (200 mL) was added, the mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure and dried in vacuo to yield 6.0 g of a white solid.

[0419] MS (ESI), m / z, 205.20 [M-55] + .

[0420] Synthesis of compound 8-3

[0421] To a CH2Cl2 solution (50 mL) of 1-3 (4.0 g, 6.44 mmol, 1.0 eq) at 25°C were added 8-2 (5.0 g, 19.3 mmol, 3.0 eq), DCC (4.6 g, 22.54 mmol, 3.5 eq), and DMAP (1.2 g, 9.66 mmol, 1.5 eq) in that order. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was filtered, and the filtrate was concentrated under reduced pressure. Purification was performed on a silica gel column (PE / EA = 1:1) as the eluent to obtain 6.0 g of the pure product.

[0422] MS (ESI), m / z, 817.20 [M-55] + .

[0423] Synthesis of compound 8-4

[0424] To a CH2Cl2 solution (10 mL) of 8-3 (2.5 g, 2.86 mmol, 1.0 eq) was added dropwise TFA (10 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the product as a yellow solid (2.5 g).

[0425] MS (ESI), m / z, 773.20 [M+H] + .

[0426] Synthesis of compound 8-5

[0427] At 25°C under nitrogen, dissolve the crude product 1-6 (800 mg, 0.285 mmol, 1.0 eq) in CH2Cl2 (20 mL). Cool to 0°C, and dropwise add a solution of 8-4 (1.24 g, 1.425 mmol, 5.0 eq) in DCM (5 mL) and triethylamine (288 mg, 2.85 mmol, 10 eq) to the reaction mixture. After the additions are complete, remove the ice-water bath and stir at room temperature for 1 h. After completion of the reaction, concentrate under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (150 mL x 3). Combine the organic phases, dry over anhydrous Na2SO4, and concentrate under reduced pressure. The crude product is then passed through a normal phase column, initially using an EA / MeOH = 1:0-10:1 elution system, then switching to a DCM / MeOH = 15:1-13:1-11:1 elution system to obtain 730 mg of crude product, which is directly used in the next step.

[0428] Synthesis of compound 8

[0429] To a THF solution (20 mL) of 8-5 (686 mg, 0.19 mmol, 1.0 eq) was added a pyridine solution of hydrogen fluoride (70%, 315 mg, 2.22 mmol, 12 eq) at 0°C. The mixture was stirred at room temperature for 2 h. A small amount of saturated aqueous NaHCO₃ was added to neutralize the pH to neutral. The mixture was concentrated under reduced pressure, and the crude product was purified on a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 300 mg of the pure product.

[0430] MS (ESI), m / z, 607.0 [DAG-OH] + .697.1[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0431] 1H NMR(400MHz,Chloroform-d)δ7.96(d,J=9.0Hz,1H),7.48–7.31(m,2H),7.19(d,J=14.2Hz,1H),7.03(d,J=7. 3Hz,1H),5.68(dd,J=17.1,3.6Hz,1H),5.40(dd,J=17.1,2.7Hz,1H),5.27–5.04(m,3H),4.60-4.44(m,1H),4. 40–3.90(m,5H),3.87-3.77(m,1H),3.77-3.42(m,219H),3.38(s,3H),3.33-2.90(m,2H),2.72-2.57(m,1H), 2.52–1.93(m,6H),1.68-1.50(m,8H),1.40-1.20(m,79H),0.97(td,J=7.4,5.3Hz,2H),0.88(t,J=6.7Hz,6H).

[0432] Example 9: Synthesis of Compound 9

[0433] Synthesis of compound 9

[0434] At room temperature of 25°C and under N2 protection, CH2Cl2 (20 mL) was added to the crude product of 1-6 (800 mg, 0.29 mmol, 1.0 eq) to dissolve it. The temperature was lowered to 0°C, and a DCM solution (5 mL) of 9-SM (prepared according to the synthesis method of Example 10114A in patent WO2023078464A1) (1.3 g, 1.425 mmol, 5.0 eq) and triethylamine (288 mg, 2.85 mmol, 10 eq) were added dropwise to the reaction solution. After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 1 h. After the reaction was complete, the mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed with water 3 times (150 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was passed through a normal phase column, first using an elution system of EA / MeOH = 1:0-10:1, and then switched to an elution system of DCM / MeOH = 15:1-13:1-11:1 to obtain 660 mg of a white solid.

[0435] MS (ESI), m / z, 607.0 [DAG-OH] + .791.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0436] Example 10: Synthesis of Compound 10

[0437] Synthesis of compounds 10-1 and 10-2

[0438] To a solution of cysteamine hydrochloride (1.50 g, 13.2 mmol, 1.0 eq) in methanol (18 mL) under an ice-water bath and nitrogen atmosphere was added triethylamine (1.85 mL, 26.4 mmol, 2.0 eq) and 2-hydroxyethyl disulfide (1.50 g, 13.2 mmol, 1.0 eq) in dichloromethane. The mixture was stirred at room temperature for 16 h. The solution from 10-1 was cooled to 0°C and di-tert-butyl dicarbonate (4.32 g, 19.8 mmol, 1.5 eq) was added. The mixture was stirred at room temperature for 4 h. LCMS analysis was performed and the reaction was terminated. The reaction solution was directly concentrated and purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain the compound as a colorless oil (0.9 g, 26.9% yield).

[0439] MS (ESI), m / z, 154.0 [M-100] + .

[0440] Synthesis of compound 10-3

[0441] To a CH2Cl2 solution (25 mL) of 10-2 (1.90 g, 7.5 mmol, 1.5 eq) was added Et3N (757 mg, 7.5 mmol, 1.5 eq) and di(p-nitrobenzene) carbonate (2.28 g, 7.5 mmol, 1.5 eq) at 0°C. The reaction was stirred for 2 h. DMAP (610 mg, 5.00 mmol, 1.0 eq) and 1-3 (3.15 g, 5.00 mmol, 1.0 eq) were added to the reaction solution. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction solution was poured into saturated aqueous ammonium chloride and washed thoroughly (150 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, and passed through a silica gel column with PE / EA = 1:1 as the eluent to obtain 2.1 g of the product.

[0442] MS (ESI), m / z, 909.1 [M+H] + .

[0443] Synthesis of compound 10-4

[0444] A formic acid solution (20 mL) of 10-3 (1.7 g, 1.80 mmol, 1.0 eq) was stirred at 0°C for 2 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure and purified on a C18 reverse phase column (CH3CN / H2O = 5-75%) to obtain 1.2 g of a yellow solid.

[0445] MS (ESI), m / z, 809.0 [M+H] + .

[0446] Synthesis of compound 10-5

[0447] At room temperature (25°C) under N₂ protection, dissolve the crude product 1-6 (800 mg, 0.285 mmol, 1.0 eq) in CH₂Cl₂ (20 mL). Cool to 0°C, and dropwise add a solution of 10-4 (1.1 g, 1.425 mmol, 5.0 eq) in DCM (5 mL) and triethylamine (288 mg, 2.85 mmol, 10 eq) to the reaction mixture. After the additions are complete, remove the ice-water bath and stir at room temperature for 1 h. After completion of the reaction, concentrate under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (150 mL x 3). Combine the organic phases, dry over anhydrous Na₂SO₄, and concentrate under reduced pressure. The crude product is then passed through a normal phase column, initially using an EA / MeOH = 1:0-10:1 elution system, then switching to a DCM / MeOH = 15:1-13:1-11:1 elution system to yield 960 mg of a white solid.

[0448] Synthesis of compound 10

[0449] To a THF solution (20 mL) of 10-5 (700 mg, 0.19 mmol, 1.0 eq) was added a pyridine solution of hydrogen fluoride (70%, 320 mg, 2.22 mmol, 12.0 eq) at 0°C. The mixture was stirred at room temperature for 2 h. A small amount of saturated aqueous NaHCO₃ was added to neutralize the pH to neutral. The mixture was concentrated under reduced pressure, and the crude product was passed through a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 370 mg of the pure product.

[0450] MS (ESI), m / z, 606.7 [DAG-OH] + .706.3[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0451] Example 11: Synthesis of Compound 11

[0452] Synthesis of compound 11-1

[0453] To a solution of CA-4 (1.67 g, 5.27 mmol, 1.0 eq) in CH2Cl2 (40 mL) at 25°C were added 7-2 (1.50 g, 6.46 mmol, 1.2 eq), DMAP (1.93 g, 15.7 mmol, 3.0 eq), and EDCI (3.0 g, 15.70 mmol, 3.0 eq) in sequence. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction solution was poured into a saturated aqueous NH4Cl solution and washed thoroughly (150 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, dried in vacuo, and purified on a silica gel column using PE / EA = 1:1 as the eluent to obtain 2.4 g of the pure product.

[0454] MS (ESI), m / z, 431.0 [M-100] + .

[0455] Synthesis of compound 11-2

[0456] To a CH2Cl2 solution (24 mL) of 11-1 (2.1 g, 3.96 mmol, 1.0 eq) was added dropwise TFA (4 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the product as a white solid (2.1 g).

[0457] MS (ESI), m / z, 431.0 [M+H] + .

[0458] Synthesis of compound 11

[0459] At room temperature (25°C) under N₂ protection, dissolve the crude product of 1-6 (755 mg, 0.269 mmol, 1.0 eq) in CH₂Cl₂ (10 mL). Cool to 0°C, and dropwise add a solution of 11-2 (750 mg, 1.38 mmol, 5.0 eq) in DCM (2 mL) and triethylamine (271 mg, 2.69 mmol, 10 eq) to the reaction mixture. After the additions were complete, remove the ice-water bath and stir at room temperature for 2 h. TLC confirmed the reaction was complete. The reaction mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed three times with water (150 mL x 3). The combined organic phases were dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The crude product was then passed through a normal phase column, initially using an EA / MeOH = 1:0-10:1 elution system, then switching to a DCM / MeOH = 15:1-13:1-11:1 elution system to obtain the compound as a white solid (610 mg).

[0460] MS (ESI), m / z, 607.0 [DAG-OH] + .671.0[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0461] 1 H NMR(400MHz,Chloroform-d)δ7.18–7.12(m,1H),7.04–6.98(m,1H),6.87(d,J=8.6Hz,1H),6.49 (d,J=5.4Hz,3H),5.90–5.83(m,1H),5.29–5.21(m,1H),4.37–4.32(m,3H),4.23–4.04(m,8H),3 .84(s,3H),3.81(s,3H),3.71(s,6H),3.68–3.65(m,193H),3.58–3.48(m,3H),3.51–3.42(m,4H ),3.40(s,3H),2.36–2.30(m,4H),1.64–1.59(m,5H),1.32-1.27(m,70H),0.90(t,J=6.7Hz,6H).

[0462] Example 12: Synthesis of Compound 12

[0463] Synthesis of compound 12-1

[0464] To a solution of paclitaxel (3.8 g, 4.45 mmol, 1.0 eq) in CH2Cl2 (60 mL) at 25°C were added 7-2 (1.25 g, 5.35 mmol, 1.2 eq), DMAP (1.63 g, 13.35 mmol, 3.0 eq), and EDCI (2.56 g, 13.35 mmol, 3.0 eq) in sequence. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction solution was poured into saturated aqueous ammonium chloride and washed thoroughly (150 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, dried in vacuo, and purified on a silica gel column using PE / EA = 1:2 to 1:3 as the eluent to obtain 3.0 g of the pure product.

[0465] MS (ESI), m / z, 967.6 [M-100] + .

[0466] Synthesis of compound 12-2

[0467] A formic acid solution (10 mL) of compound 12-1 (2.0 g, 1.87 mmol, 1.0 eq) was stirred at 0°C for 6 h. After completion, the reaction solution was concentrated to 3 mL at 0°C. Tert-butyl methyl ether (100 mL) was added dropwise to slurry the mixture, resulting in the precipitation of a large amount of white solid. The filter cake was filtered and thoroughly vacuum-dried to yield 1.2 g of a pure white solid.

[0468] MS (ESI), m / z, 967.6 [M+H] + .

[0469] Synthesis of compound 12

[0470] At room temperature (25°C) under N2 protection, dissolve the crude product 1-6 (755 mg, 0.27 mmol, 1.0 eq) in CH2Cl2 (10 mL). Cool to 0°C, and dropwise add a solution of 12-2 (1.49 g, 1.38 mmol, 5.0 eq) in DCM (2 mL) and triethylamine (271 mg, 2.69 mmol, 10 eq) to the reaction mixture. After the addition is complete, remove the ice-water bath and stir at room temperature for 2 h. TLC confirms the reaction is complete. The reaction mixture is concentrated under reduced pressure, dissolved in DCM (100 mL), and washed three times with saturated brine (150 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was passed through a normal phase column, first using an elution system of EA / MeOH = 1:0-10:1, and then switched to an elution system of DCM / MeOH = 95:5-93:7-92:8-89:11 to obtain 710 mg of the product.

[0471] MS (ESI), m / z, 606.8 [DAG-OH] + .805.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0472] Example 13: Synthesis of Compound 13

[0473] Synthesis of compound 13-1

[0474] 10-2 (300 mg, 1.18 mmol, 1.0 eq) was dissolved in dichloromethane (20 mL) at room temperature. Triethylamine (120 mg, 1.18 mmol, 1.0 eq) and di(p-nitrobenzene) carbonate (360 mg, 1.18 mmol, 1.0 eq) were added under nitrogen. The mixture was stirred at room temperature for 16 h. 4-Dimethylaminopyridine (144.6 mg, 1.18 mmol, 1.0 eq) and paclitaxel (1010 mg, 1.18 mmol, 1.0 eq) were added. The mixture was stirred at room temperature for 2 h. The reaction was terminated by LCMS analysis. The product was concentrated under reduced pressure and purified on a reverse-phase column (A = TFA (0.1% + H2O), B = acetonitrile)). The product was then lyophilized to afford 13-1 as a white solid (0.8 g, 59.7% yield).

[0475] MS (ESI), m / z, 1133.0 [M+H] + .

[0476] Synthesis of compound 13-2

[0477] Dissolve 13-1 (800 mg, 0.71 mmol, 1.0 eq) in formic acid (10 mL) at room temperature. Stir under nitrogen for 2 h at room temperature. LCMS analysis was performed. Terminate the reaction. Concentrate under reduced pressure, separate by reverse phase separation (A = TFA (0.1% + H2O), B = acetonitrile), and lyophilize to afford 13-2 as a white solid (0.4 g, 54.8% yield).

[0478] Synthesis of compound 13

[0479] At room temperature (25°C) under N₂ protection, dissolve the crude product 1-6 (800 mg, 0.29 mmol, 1.0 eq) in CH₂Cl₂ (20 mL). Cool to 0°C, and dropwise add a DCM solution (5 mL) of 13-2 (1.47 g, 1.43 mmol, 5.0 eq) and triethylamine (288 mg, 2.85 mmol, 10 eq) to the reaction mixture. After the addition is complete, remove the ice-water bath and stir at room temperature for 1 h. After completion of the reaction, concentrate under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (150 mL x 3). Combine the organic phases, dry over anhydrous Na₂SO₄, and concentrate under reduced pressure. The crude product is then passed through a normal phase column, initially using an EA / MeOH = 1:0-10:1 elution system, then switching to a DCM / MeOH = 15:1-13:1-11:1 elution system to yield 0.97 g of a white solid.

[0480] MS (ESI), m / z, 606.8 [DAG-OH] + .821.6[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0481] Example 14: Synthesis of Compound 14

[0482] Synthesis of compound 14-1

[0483] To a THF solution (60 mL) of N-Boc-L-lysine (3.0 g, 12.2 mmol, 1.0 eq) at 25°C was added Fmoc-OSu (4.5 g, 13.4 mmol, 1.1 eq) and DIPEA (1.7 g, 13.4 mmol, 1.1 eq). The mixture was stirred at room temperature for 4 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The crude product was purified on a C18 column with CH3CN:H2O = 5-95% to afford 4.0 g of a pure white solid.

[0484] MS (ESI), m / z, 369.0 [M-100] + .

[0485] Synthesis of compound 14-2

[0486] To a DMF solution of 14-1 (1.0 g, 2.14 mmol, 1.0 eq) (10 mL) were added 9-SM (2.14 g, 2.35 mmol, 1.1 eq), HATU (1.22 g, 3.21 mmol, 1.5 eq), and DIPEA (550 mg, 4.28 mmol, 2.0 eq) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the DMF was concentrated under reduced pressure using an oil pump. The crude product was purified on a C18 column with CH3CN:H2O = 5-95% to obtain 2.9 g of a pure white solid.

[0487] MS (ESI), m / z, 1361.6 [M-100] + .

[0488] Synthesis of compound 14-3

[0489] To a CH2Cl2 solution (5 mL) of 14-2 (1.0 g, 7.34 mmol, 1.0 eq) was added dropwise TFA (5 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to yield 1.0 g of the compound as a white solid.

[0490] MS (ESI), m / z, 1361.60 [M-100] + .

[0491] Synthesis of compound 14-4

[0492] At room temperature (25°C), to a DMF solution (4 mL) of 14-4 (700 mg, 0.5 mmol, 1.0 eq) were added 9-SM2 (prepared according to the synthesis method of Example 10114-1K in patent WO2016046574A1, 1.22 g, 0.60 mmol, 1.2 eq), HATU (342 mg, 0.9 mmol, 1.8 eq), and DIPEA (155 mg, 1.2 mmol, 2.4 eq). After addition, the mixture was stirred at room temperature for 2 h. The reaction solution was poured into water (100 mL) and extracted with EA (70 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified by normal phase column chromatography (DCM / MeOH = 10:1) to obtain 1.2 g of pure product.

[0493] Synthesis of compound 14-5

[0494] To a DMF solution (15 mL) of 14-4 (1.1 g, 0.34 mmol, 1.0 eq) was added dropwise Et3N (3 mL) at 25°C. The mixture was stirred at room temperature for 9 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was thoroughly concentrated using an oil pump. The crude product was purified by normal phase column chromatography (DCM / MeOH / H2O = 75:22:3 to DCM / MeOH / H2O / Et3N = 75:22:3:5) to obtain 900 mg of pure product as a white solid.

[0495] Synthesis of compound 14

[0496] At room temperature (25°C) and under N₂ protection, DMF (11 mg, 0.148 mmol, 0.5 eq) was added to a CH₂Cl₂ solution (10 mL) of DSPC (CAS No. 816-94-4, 280 mg, 0.355 mmol, 1.2 eq). The temperature was lowered to 0°C, and oxalyl chloride (180 mg, 1.42 mmol, 4.8 eq) was added dropwise to the reaction mixture. After the addition was complete, the mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under N₂ protection to afford 290 mg of crude white solid 14-6, which was used directly in the next reaction.

[0497] At room temperature (25°C) and under N2 protection, CH2Cl2 (10 mL) was added to the above crude product to dissolve it. The temperature was lowered to 0°C, and a DCM solution (6 mL) of 14-5 (900 mg, 0.296 mmol, 1.0 eq) and triethylamine (143 mg, 1.42 mmol, 4.8 eq) were added dropwise to the reaction solution. After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure, dissolved in DCM (50 mL), and washed three times with saturated NaCl (200 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified by normal phase column chromatography (DCM / MeOH / H2O = 75:22:3) to obtain the compound, 480 mg.

[0498] MS (ESI), m / z, 607.0 [DAG-OH] + .826.6[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0499] Example 15: Synthesis of Compound 15

[0500] Synthesis of compound 15-1

[0501] To a CH2Cl2 solution (70 mL) of SN38 (2.0 g, 3.17 mmol, 1.0 eq) at 25°C was added Boc-β-alanine (1.5 g, 7.93 mmol, 2.5 eq), EDCI (1.81 g, 9.50 mmol, 3.0 eq), and DMAP (581 mg, 4.76 mmol, 1.5 eq) in that order. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure and purified on a silica gel column (PE / EA = 2:1 to 1:1 as eluent) to afford 2.7 g of a yellow solid.

[0502] MS (ESI), m / z, 801.60 [M+H] + .

[0503] Synthesis of compound 15-2

[0504] To a CH2Cl2 solution (20 mL) of 15-1 (2.3 g, 2.87 mmol, 1.0 eq) was added dropwise TFA (18 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the compound as a yellow solid (2.8 g).

[0505] MS (ESI), m / z, 701.80 [M+H] + .

[0506] Synthesis of compound 15-3

[0507] At 25°C under N2 protection, dissolve the crude product 1-6 (800 mg, 0.285 mmol, 1.0 eq) in CH2Cl2 (20 mL). Cool to 0°C, and dropwise add a solution of 15-2 (1.14 g, 1.425 mmol, 5.0 eq) in DCM (5 mL) and triethylamine (288 mg, 2.85 mmol, 10 eq) to the reaction mixture. After the additions are complete, remove the ice-water bath and stir at room temperature for 1 h. Monitor the reaction by TLC. Upon completion, concentrate under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (150 mL x 3). Combine the organic phases, dry over anhydrous Na2SO4, and concentrate under reduced pressure. The crude product is then passed through a normal phase column, initially using an EA / MeOH = 1:0-10:1 elution system, then switching to a DCM / MeOH = 10:0-10:1-5:1 elution system to yield 745 mg of product.

[0508] Synthesis of compound 15

[0509] To a THF solution (4 mL) of 15-3 (280 mg, 0.0848 mmol, 1.0 eq) at 0°C was added TBAF (100 μL, 0.102 mmol, 1.2 eq) and AcOH (25 mg, 0.424 mmol, 5.0 eq). The mixture was returned to room temperature and stirred for 2 h. The reaction mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed three times with water (150 mL x 2). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified on a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 60 mg of the pure product.

[0510] MS (ESI), m / z, 606.8 [DAG-OH] + .679.3[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0511] 1 H NMR(400MHz,Chloroform-d)δ(ppm)9.20(s,1H),8.04(dd,J=9.1,1.4Hz,1H),7.58–7.46(m,2H),7.14(s,1H),5.69(d d,J=17.0,2.5Hz,1H),5.59(s,1H),5.39(dd,J=17.0,4.2Hz,1H),5.20(d,J=15.2Hz,3H),4.31(dt,J=12.0,4.2Hz,1H ),4.24–3.77(m,5H),3.64(d,J=5.3Hz,171H),3.38(s,7H),3.10(q,J=7.7Hz,2H),2.82–2.59(m,3H),2.37–2.20(m,5 H),1.75–1.53(m,9H),1.51–1.32(m,8H),1.25(d,J=3.2Hz,91H),1.00(dd,J=8.3,6.2Hz,9H),0.88(t,J=6.6Hz,9H).

[0512] Example 16: Synthesis of Compound 16

[0513] Synthesis of compound 16-1

[0514] To a THF solution (100 mL) of N-tert-butyloxycarbonyl-1,3-propanediamine (10 g, 57.4 mmol, 1.05 eq) at 25°C, succinic anhydride (5.48 g, 54.7 mmol, 1.0 eq) and DIPEA (14.1 g, 109.4 mmol, 2.0 eq) were slowly added. The reaction mixture was concentrated and dried under vacuum to yield 17 g of a white solid.

[0515] MS (ESI), m / z, 274.80 [M+H] + .

[0516] Synthesis of compound 16-2

[0517] To a CH2Cl2 solution (70 mL) of 1-3 (2.0 g, 3.17 mmol, 1.0 eq) at 25°C were added 16-2 (2.2 g, 7.93 mmol, 2.5 eq), EDCI (1.81 g, 9.50 mmol, 3.0 eq), and DMAP (581 mg, 4.76 mmol, 1.5 eq) in that order. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA = 2:1 to 1:1 as eluent) to afford 1.6 g of a yellow-green solid.

[0518] MS (ESI), m / z, 886.60 [M+H] + .

[0519] Synthesis of compound 16-3

[0520] To a CH2Cl2 solution (20 mL) of 16-2 (2.3 g, 2.60 mmol, 1.0 eq) was added dropwise TFA (18 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the compound as a yellow solid (2.7 g).

[0521] MS (ESI), m / z, 786.60 [M+H] + .

[0522] Synthesis of compound 16-4

[0523] At room temperature (25°C) under N₂ protection, dissolve the crude product 1-6 (800 mg, 0.29 mmol, 1.0 eq) in CH₂Cl₂ (20 mL). Cool to 0°C, and dropwise add a solution of 16-3 (1.14 g, 1.43 mmol, 5.0 eq) in DCM (5 mL) and triethylamine (288 mg, 2.85 mmol, 10 eq) to the reaction mixture. After the additions were complete, remove the ice-water bath and stir at room temperature for 1 h. TLC indicated completion of the reaction. The product was concentrated, dissolved in DCM (100 mL), and washed three times with water (150 mL x 3). The combined organic phases were dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The crude product was then passed through a normal phase column, initially using an EA / MeOH = 1:0-10:1 elution system, then switching to a DCM / MeOH = 10:0-10:1-5:1 elution system to yield 710 mg of product.

[0524] Synthesis of compound 16

[0525] To a THF solution of 16-4 (590 mg, 0.159 mmol, 1.0 eq) (15 mL) at 0°C was added TBAF (175 μL, 0.175 mmol, 1.1 eq) and AcOH (95 mg, 1.59 mmol, 10 eq). The mixture was returned to room temperature and stirred for 2 h. The reaction mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed three times with water (150 mL x 2). The combined organic phases were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude product was purified on a silica gel column (DCM / MeOH = 15:1 to 13:1 to 11:1) to obtain 710 mg of the pure product.

[0526] MS (ESI), m / z, 606.8 [DAG-OH] + .700.1[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0527] 1H NMR(400MHz,Chloroform-d)δ(ppm)8.84(s,1H),8.08(dd,J=9.2,1.5Hz,1H),7.47(dd,J=9.1,2.6Hz,1H),7.57–7.32(m,2H),7.2 1(d,J=2.7Hz,1H),6.57-6.44(m,1H),5.90–5.56(m,2H),5.37(d,J=17.1Hz,1H),5.27-5.13(m,3H),4.37-4.26(m,1H),4.25–3.77 (m,6H),3.75–3.40(m,214H),3.38-3.30(m,4H),3.29–3.18(m,2H),3.08(d,J=7.9Hz,2H),3.00–2.75(m,4H),2.60-2.45(m,2H),2 .347-2.26(m,5H),1.67-1.45(m,9H),1.39-1.21(m,104H),1.13-1.04(m,1H),0.99(td,J=7.5,1.5Hz,3H),0.88(t,J=6.7Hz,9H).

[0528] Example 17: Synthesis of Compound 17

[0529] Synthesis of compound 17-1

[0530] To a THF solution (30 mL) of exatecan hydrochloride (500 mg, 1.06 mmol, 1.0 eq) at 25°C was added Fmoc-OSu (536 mg, 1.59 mmol, 1.5 eq) and Et3N (160 mg, 1.59 mmol, 1.5 eq). The mixture was stirred at room temperature for 4 h. After completion of the reaction, the mixture was concentrated and purified on a silica gel column (PE / EA = 1:2) to yield 560 mg of a pure white solid.

[0531] MS (ESI), m / z, 657.8 [M+H] + .

[0532] Synthesis of compound 17-2

[0533] To a CH2Cl2 solution (10 mL) of 17-1 (560 mg, 0.85 mmol, 1.0 eq) at 25°C were added 5-2 (630 mg, 2.56 mmol, 3.0 eq), EDCI (571 mg, 2.99 mmol, 3.5 eq), and DMAP (156 mg, 1.28 mmol, 1.5 eq) in that order. The mixture was stirred at room temperature for 3 h. After completion of the reaction, the mixture was washed with saturated ammonium chloride solution. The organic phase was separated and dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified on a silica gel column (PE / EA = 1:2) to afford 630 mg of a solid.

[0534] MS (ESI), m / z, 885.8 [M+H] + .

[0535] Synthesis of compound 17-3

[0536] To a CH2Cl2 solution (4 mL) of 17-2 (630 mg, 0.71 mmol, 1.0 eq) was added dropwise 4M HCl / dioxane (8 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated and dissolved in acetonitrile / methanol (1:1). Further concentration was performed to yield 630 mg of a yellow solid.

[0537] MS (ESI), m / z, 785.6 [M+H] + .

[0538] Synthesis of compound 17-4

[0539] At 25°C under nitrogen, dichloromethane (10 mL) was added to the crude product 1-6 (755 mg, 0.27 mmol, 1.0 eq) to dissolve it. The temperature was lowered to 0°C, and a dichloromethane solution of 17-3 (663 mg, 0.81 mmol, 3.0 eq) was added to the reaction solution, followed by the dropwise addition of triethylamine (271 mg, 2.69 mmol, 10 eq). After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 2 h. TLC confirmed the reaction was complete. The reaction solution was concentrated, dissolved in dichloromethane (100 mL), and washed three times with water (150 mL x 3). The combined organic phases were dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified by normal phase column chromatography (EA / MeOH = 1:0-10:1, followed by DCM / MeOH = 95:5-94:6-10:1) to yield 700 mg of crude product. The crude product was then purified by reverse phase column ((THF / MeOH=3:1):H2O=5-90%) to obtain 440 mg of pure product.

[0540] Synthesis of compound 17

[0541] To a solution of 17-4 (370 mg, 0.10 mmol, 1.0 eq) in DMF (10 mL) was added Et3N (2 mL) at 25°C. The mixture was stirred at room temperature for 4 h. After completion of the reaction, the mixture was concentrated under reduced pressure at room temperature. The crude product was purified by normal phase column chromatography (DCM / MeOH / H2O = 10:1:0 to 75:22:3) to obtain 118 mg of the product. HPLC purity was 95.5%.

[0542] MS (ESI), m / z, 606.8 [DAG-OH] + .704.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0543] 1 H NMR(400MHz,Chloroform-d)δ7.73(d,J=10.5Hz,1H),7.37-7.20(m,1H),5.72-5 .61(m,2H),5.43-5.35(m,2H),5.20-5.18(m,1H),4.46-3.67(m,8H),3.60-3.50 (m,191H),3.40(s,3H),3.36–3.06(m,1H),2.44(s,2H),2.39–2.14(m,8H),1.67 -1.60(m,6H),1.40-1.27(m,77H),1.01(t,J=7.4Hz,3H),0.90(t,J=6.7Hz,6H).

[0544] Example 18: Synthesis of Compound 18

[0545] Synthesis of compound 18-1

[0546] To a THF solution (30 mL) of 18-SM (prepared according to the synthetic method of Example 1 in WO2023030364A1, 600 mg, 1.30 mmol, 1.0 eq) was added Fmoc-OSu (661 mg, 1.96 mmol, 1.5 eq) and Et3N (198 mg, 1.96 mmol, 1.5 eq) at 25°C. The mixture was stirred at room temperature for 4 h. After completion of the reaction, the mixture was purified on a silica gel column (PE / EA = 2:1 to 1:2) to obtain 810 mg of a pure light yellow-green solid.

[0547] MS (ESI), m / z, 645.80 [M+H] + .

[0548] Synthesis of compound 18-2

[0549] To a CH2Cl2 solution (15 mL) of 18-1 (810 mg, 1.26 mmol, 1.0 eq) at 25°C were added 5-2 (927 mg, 3.77 mmol, 3.0 eq), EDCI (856 mg, 4.48 mmol, 3.5 eq), and DMAP (234 mg, 1.92 mmol, 1.5 eq) in that order. The mixture was stirred at room temperature for 3 h. After completion of the reaction, saturated ammonium chloride was added for washing. The organic phase was separated and dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified on a silica gel column (PE / EA = 1:2) to afford 1.2 g of a solid.

[0550] MS (ESI), m / z, 873.8 [M+H] + .

[0551] Synthesis of compound 18-3

[0552] To a CH2Cl2 solution (6 mL) of compound 18-2 (1.1 g, 1.26 mmol, 1.0 eq) was added dropwise TFA (1 mL) at 25°C. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated and dried under vacuum to yield 1.1 g of a yellow solid.

[0553] MS (ESI), m / z, 773.80 [M+H] + .

[0554] Synthesis of compound 18-4

[0555] At room temperature (25°C) under N₂ protection, dissolve the crude product 1-6 (1.51 g, 0.54 mmol, 1.0 eq) in CH₂Cl₂ (20 mL). Cool to 0°C, and add a dichloromethane solution of 18-3 (1.12 g, 1.34 mmol, 2.5 eq) to the reaction mixture, followed by the dropwise addition of triethylamine (543 mg, 5.38 mmol, 10 eq). After the addition is complete, remove the ice-water bath and stir at room temperature for 2 h. TLC confirms the reaction is complete. Concentrate the reaction mixture, dissolve it in dichloromethane (100 mL), and wash with water three times (150 mL x 3). Combine the organic phases, dry over anhydrous Na₂SO₄, and concentrate under reduced pressure. Purify the crude product using a normal phase column (DCM / MeOH = 9:1) to yield 1.37 g of crude product. The crude product was then purified using a reverse phase C8 column (THF / MeOH=3:1):H2O=5-90%) to obtain 1.06 g of pure product.

[0556] Synthesis of compound 18

[0557] To a solution of 18-4 (1.05 g, 0.31 mmol, 1.0 eq) in DMF (10 mL) was added Et3N (2 mL) at 25°C. The mixture was stirred at room temperature for 4 h. After completion of the reaction, the mixture was concentrated under reduced pressure, and the crude product was purified by normal phase column chromatography (DCM / MeOH / H2O = 10:1:0 to 75:22:3) to obtain 118 mg of the product.

[0558] MS (ESI), m / z, 606.8 [DAG-OH] + .701.3[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0559] 1 H NMR(400MHz,Chloroform-d)δ8.11(d,J=8.0Hz,1H),7.84(d,J=10.3Hz,1H), 7.40-7.30(m,1H),5.87–5.63(m,2H),5.53–5.15(m,3H),4.51–3.76(m,9H), 3.74-3.48(m,205H),,3.40(s,3H),2.56(s,2H),2.34–2.24(m,5H),1.68–1. 54(m,8H),1.46–1.19(m,72H),1.01(t,J=7.4Hz,3H),0.90(t,J=6.7Hz,6H).

[0560] Example 19: Synthesis of Compound 19

[0561] Synthesis of compound 19-1

[0562] To a CH₂Cl₂ solution (25 mL) of 10-2 (1.3 g, 5.0 mmol, 1.5 eq) at 0°C, Et₃N (510 mg, 5.0 mmol, 1.5 eq) and di(p-nitrobenzene) carbonate (1.52 g, 5.0 mmol, 1.5 eq) were added sequentially. The reaction was continued with stirring at this temperature for 2 h. DIPEA (494 mg, 3.83 mmol, 3.0 eq) and a solution of exatecan hydrochloride (600 mg, 1.28 mmol, 1.0 eq) in DMF (15 mL) were then added. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was poured into water (100 mL). The mixture was extracted with EA (100 mL x 3). The organic phases were combined, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The crude product was passed through a normal phase column using a PE / EA = 1:1 to 1:2 eluent to yield 720 mg of a light yellow-green solid.

[0563] MS (ESI), m / z, 715.8 [M+H] + .

[0564] Synthesis of compound 19-2

[0565] A solution of 19-1 (700 mg, 0.98 mmol, 1.0 eq) in formic acid (10 mL) was stirred at 0°C for 6 h. After completion, the reaction solution was concentrated to 3 mL at 0°C. MTBE (30 mL) was added dropwise to slurry the mixture, resulting in the precipitation of a large amount of solid. The filter cake was filtered and thoroughly dried under vacuum to yield 720 mg of solid.

[0566] MS (ESI), m / z, 615.8 [M+H] + .

[0567] Synthesis of compound 19

[0568] At 25°C under nitrogen, dissolve the crude product 1-6 (755 mg, 0.269 mmol, 1.0 eq) in CH2Cl2 (15 mL). Cool to 0°C, and sequentially add 19-2 (710 mg, 1.076 mmol, 4.0 eq) and triethylamine (271 mg, 2.69 mmol, 10 eq) to the reaction mixture. Remove the ice-water bath and stir at room temperature for 2 h. TLC confirms the reaction is complete. Concentrate the reaction mixture under pressure, dissolve it in DCM (100 mL), and wash with water three times (150 mL x 3). Combine the organic phases, dry over anhydrous Na2SO4, and concentrate under reduced pressure. Pass the crude product through a normal phase column, initially eluting with EA / MeOH = 1:0-10:1, then switching to DCM / MeOH = 15:1-13:1-11:1, to yield 330 mg of product.

[0569] MS (ESI), m / z, 606.8 [DAG-OH] + .717.0[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0570] 1H NMR(400MHz,Chloroform-d)δ7.67(d,J=10.5Hz,1H),7.62(s,1H),6.70-6.54(m,1H),5.75-5 .71(m,2H),5.50–5.11(m,6H),4.65–4.53(m,1H),4.48–3.75(m,11H),3.73-3.55(m,187H),3 .53-3.46(m,2H),3.40(s,3H),3.35-3.20(m,4H),3.18-3.02(m,4H),2.93-2.81(m,2H),2.43 (s,3H),2.38–2.22(m,9H),1.39-1.20(m,82H),1.07(t,J=7.4Hz,3H),0.90(t,J=6.7Hz,9H).

[0571] Example 20: Synthesis of Compound 20

[0572] Synthesis of compound 20-1

[0573] To a DCM solution of 10-2 (2.53 g, 10.0 mmol, 1.0 eq) (50 mL) at 0°C were added di-p-nitrophenyl carbonate (3.19 g, 10.5 mmol, 1.05 eq) and Et3N (1.11 g, 11 mmol, 1.1 eq). After addition, the mixture was returned to room temperature and stirred for 2 h. Upon completion of the reaction, half of the reaction mixture was used in the next reaction. MS (ESI), m / z, 318.8.

[0574] To a solution of 18-SM (600 mg, 1.30 mmol, 1.0 eq) in DMF (200 mL) at 25°C were added the reaction mixture of the intermediate 10-2 obtained in the previous step, DIPEA (503 mg, 3.9 mmol, 3.0 eq), DMAP (476 mg, 3.90 mmol, 3.0 eq), and pyridine (2 mL). The mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was poured into water (800 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic phases were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude product was purified by a normal phase column using a PE / EA = 1:1-1:2 eluent to yield 1.0 g of crude product. The crude product was further purified by a reverse phase column (CH3CN:H2O = 5-90%) to afford 640 mg of a white solid.

[0575] MS (ESI), m / z, 702.80 [M+H] + .

[0576] Synthesis of compound 20-2

[0577] A formic acid solution (10 mL) of 20-1 (620 mg, 0.88 mmol, 1.0 eq) was stirred at 0°C for 6 h. After completion, the reaction solution was concentrated to 3 mL at 0°C. MTBE (30 mL) was added dropwise to slurry the mixture, resulting in the precipitation of a large amount of white solid. The filter cake was filtered and thoroughly dried under vacuum to yield 670 mg of a solid.

[0578] MS (ESI), m / z, 602.8 [M+H] + .

[0579] Synthesis of compound 20

[0580] At 25°C under nitrogen, dissolve the crude product 1-6 (755 mg, 0.27 mmol, 1.0 eq) in CH2Cl2 (20 mL). Cool to 0°C, and sequentially add 20-2 (696 mg, 1.08 mmol, 5.0 eq) and triethylamine (271 mg, 2.69 mmol, 10 eq) to the reaction mixture. After the addition is complete, remove the ice-water bath and stir at room temperature for 1 h. TLC confirms the reaction is complete. Concentrate the reaction mixture under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (150 mL x 3). Combine the organic phases, dry over anhydrous Na2SO4, and concentrate under reduced pressure. The crude product is then passed through a normal phase column, initially eluting with EA / MeOH = 1:0-10:1, then switching to DCM / MeOH = 15:1-13:1-11:1, to yield 310 mg of product.

[0581] MS (ESI), m / z, 607.0 [DAG-OH] + .714.0[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0582] 1 H NMR(400MHz,Chloroform-d)δ8.15-8.05(m,1H),7.82-7.74(m,1H),7.64(d,J=11.1Hz,1H ),6.44-6.35(m,1H),5.74-5.70(m,1H),5.62–5.23(m,3H),4.45–4.07(m,4H),3.87–3.81 (m,1H),3.75-3.45(m,180H),3.40(s,3H),3.04–2.85(m,4H),2.57(s,3H),2.41-2.27(m, 5H),1.71–1.52(m,7H),1.38–1.21(m,72H),1.06(t,J=7.4Hz,3H),0.90(t,J=6.7Hz,6H).

[0583] Example 21: Synthesis of Compound 21

[0584] Synthesis of compound 21-1

[0585] To a CH2Cl2 solution (15 mL) of 21-SM (prepared according to the synthetic method of Example 40 in patent WO2018202202A1, 680 mg, 1.26 mmol, 1.0 eq) was added 5-2 (927 mg, 3.77 mmol, 3.0 eq), EDCI (856 mg, 4.48 mmol, 3.5 eq) and DMAP (234 mg, 1.92 mmol, 1.5 eq) in sequence at room temperature (25 ° C). The mixture was stirred at room temperature for 3 h. After the reaction was completed, saturated ammonium chloride was added for washing, the liquid was separated, and the organic phase was dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified on a silica gel column (PE / EA = 1:2) to obtain 1.0 g of a solid.

[0586] MS (ESI), m / z, 750.7 [M+H] + .

[0587] Synthesis of compound 21-2

[0588] To a CH2Cl2 solution (6 mL) of compound 21-1 (0.8 g, 1.07 mmol, 1.0 eq) was added dropwise TFA (1 mL) at 25°C. The mixture was stirred at room temperature for 1 h. After completion of the reaction, the reaction mixture was concentrated and dried under vacuum to yield 0.7 g of a yellow solid.

[0589] MS (ESI), m / z, 650.80 [M+H] + .

[0590] Synthesis of compound 21

[0591] At 25°C under N2 protection, crude 1-6 (1.02 g, 0.37 mmol, 1.0 eq) was dissolved in CH2Cl2 (15 mL). The temperature was lowered to 0°C, and a dichloromethane solution of 21-2 (0.7 g, 0.91 mmol, 2.5 eq) was added to the reaction solution, followed by the dropwise addition of triethylamine (370 mg, 3.64 mmol, 10 eq). After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 2 h. TLC confirmed the reaction was complete. The reaction solution was concentrated, dissolved in dichloromethane (80 mL), and washed three times with water (100 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified using a normal phase column (DCM / MeOH = 9:1) and then a reverse phase C8 column (THF / MeOH = 3:1): 5-90% H2O) to yield 0.8 g of pure product.

[0592] MS (ESI), m / z, 606.8 [DAG-OH] + .725.7[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0593] Example 22: Synthesis of Compound 22

[0594] Synthesis of compound 22-1

[0595] To a 20 mL solution of DOPE-PEG2K (1.0 g, 0.36 mmol, 1.0 eq) in CH2Cl2 was added DMF (0.1 mL) in an ice-water bath at 5°C under N2 protection, followed by the dropwise addition of oxalyl chloride (0.5 g, 3.6 mmol, 10.0 eq). After the addition was complete, the mixture was stirred at room temperature for 3 h. After completion of the reaction, the mixture was concentrated under reduced pressure under N2 protection to yield 1.0 g of a crude white solid, which was used directly in the next reaction.

[0596] Synthesis of compound 22

[0597] To a solution of crude product 22-1 (0.5 g, 0.18 mmol, 1.0 eq) in CH2Cl2 (10 mL) in an ice-water bath at 5°C under N2 protection was added dropwise a solution of 1-5 (0.78 g, 0.89 mmol, 5.0 eq) in DCM (5 mL) and triethylamine (180 mg, 1.79 mmol, 15.0 eq). After the addition was complete, the reaction mixture was warmed to room temperature and stirred for 2 h. The reaction mixture was diluted with DCM (30 mL), washed with salt and water, and separated. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to yield 0.6 g of crude oil. The crude product was dissolved in THF (10 mL), and TBAF (1.0 M in THF, 0.5 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 2 h, and the reaction solution was quenched with water. The mixture was extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt, washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was first purified by silica gel column (DCM / MeOH = 9:1) and then by neutral alumina column to obtain compound 22, 250 mg, as a white solid.

[0598] MS (ESI), m / z, 602.6 [DAG-OH] + .693.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0599] General Synthesis Method A of Phospholipid PEG:

[0600] General Synthesis Method of Compound A-1

[0601] Under an ice-water bath and nitrogen, a solution of phosphorus oxychloride (1.0 eq) was added dropwise to a THF solution of alcohol R-OH and triethylamine (1.0 eq), followed by stirring at room temperature for 4 hours. The reaction mixture was cooled in an ice-water bath and a THF solution of N-(tert-butyloxycarbonyl)ethanolamine (1.5 eq) and triethylamine (3.0 eq) was added dropwise. The reaction mixture was stirred for 2 hours and then at room temperature for 12 hours. The mixture was then quenched with water, concentrated, and acidified to pH = 1 by adding 1% HCl. The aqueous phase was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by flash column chromatography (eluted with MeOH / CH2Cl2) to obtain compound A-1.

[0602] General Synthesis Method of Compound A-2

[0603] A-1 was dissolved in dichloromethane and TFA (5 / 1, v / v), the reaction solution was stirred at room temperature for 1-12 hours, concentrated, and the residue was separated and purified by column chromatography (eluted with MeOH / CH2Cl2 system) to obtain compound A-2.

[0604] General Synthesis Method of Compound A-3

[0605] A-2 (1.0 eq) was dissolved in dichloromethane and triethylamine (5.0 eq), and a dichloromethane solution of mPEG-SC (1.0 eq) was added dropwise. The reaction solution was stirred at room temperature for 1-12 h, concentrated, and the residue was separated and purified by column chromatography (eluted with MeOH / CH2Cl2 system) to obtain compound A-3.

[0606] Phospholipid-coupled drug synthesis method B:

[0607] General Synthesis Method of Compound B-1

[0608] To a solution of A-3 (1.0 eq) in CH2Cl2, add DMF (5.0 eq) and then dropwise add oxalyl chloride (10.0 eq) in an ice-water bath at 5°C under N2 protection. After the addition is complete, stir at room temperature for 1-3 hours. After the reaction is complete, concentrate under reduced pressure under N2 protection to obtain crude product B-1, which is used directly in the next step.

[0609] General synthetic methods for compounds

[0610] To a CH2Cl2 solution of crude B-1 (1.0 eq) in an ice-water bath at 5°C under N2 protection was added dropwise a CH2Cl2 solution of an amine drug (3.0-5.0 eq) and triethylamine (10-15.0 eq). After the addition was complete, the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt, washed with water, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (eluted with MeOH / CH2Cl2) to obtain the compound. If necessary, purification was performed by reverse-phase column chromatography (eluted with MeOH / 0.05% TFA in water).

[0611] Example 23: Synthesis of Compound 23

[0612] Synthesis of compound 23-4

[0613] Phospholipid compound 23-3 was synthesized using raw materials 1,3-stearyl glyceryl and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 23-4 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0614] MS (ESI), m / z, 606.8 [DAG-OH] + .753.1[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0615] Synthesis of compound 23

[0616] Compound 23-4 (0.35 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 0.5 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 5 h, and the reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 23, 160 mg, as a white solid with an HPLC purity of 93.2%.

[0617] MS (ESI), m / z, 606.8 [DAG-OH] + .693.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0618] Example 24: Synthesis of Compound 24

[0619] Synthesis of compound 24-2

[0620] Phospholipid compound 24-1 was synthesized using raw materials 1,2-dipalmitoylglycerol and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 24-2 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0621] Synthesis of compound 24

[0622] Compound 24-2 (0.75 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 0.5 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 4 h, and the reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 24, 560 mg, as a white solid with an HPLC purity of 98.6%.

[0623] MS (ESI), m / z, 550.8 [DAG-OH] + .693.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is diglyceride.

[0624] Example 25: Synthesis of Compound 25

[0625] Synthesis of compound 25-2

[0626] Phospholipid compound 25-1 was synthesized using the starting material 25-SM (CAS: 133831-25-1, commercially available) and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 25-2 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0627] Synthesis of compound 25

[0628] Compound 25-2 (0.6 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 1.0 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 6 h, and the reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt, washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was first separated and purified by silica gel column chromatography (DCM / MeOH system) and then separated and purified by reverse phase column chromatography (MeOH / 0.05% TFA water system) to obtain compound 25, 320 mg, as a white solid with an HPLC purity of 89.6%.

[0629] MS (ESI), m / z, 694.5 [DAG-OH] +.693.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0630] Example 26: Synthesis of Compound 26

[0631] Synthesis of compound 26-2

[0632] Phospholipid compound 26-1 was synthesized using the starting materials glycerol diarachidate and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 26-2 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0633] Synthesis of compound 26

[0634] Compound 26-2 (0.50 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 0.5 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 2 h, quenched with water, and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 26, 230 mg, as a light yellow solid with an HPLC purity of 96.3%.

[0635] MS (ESI), m / z, 662.8 [DAG-OH] + .693.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0636] Example 27: Synthesis of Compound 27

[0637] Synthesis of compound 27-2

[0638] Phospholipid compound 27-1 was synthesized using the starting material 27-SM (CAS: 1169768-28-8, commercially available) and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 27-2 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0639] Synthesis of compound 27

[0640] Compound 27-2 (0.30 g) was dissolved in THF solution (5 mL), and TBAF (1.0 M, THF solution, 0.5 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 3 h, and the reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 27, 120 mg, as a white solid with an HPLC purity of 91.0%.

[0641] MS (ESI), m / z, 510.5 [ROH-OH] + .693.4[M-DAG+4Na] 4+ .ROH is an alkyl alcohol fragment.

[0642] Example 28: Synthesis of Compound 28

[0643] Synthesis of compound 28-2

[0644] Phospholipid compound 28-1 was synthesized using the starting materials 28-SM (ethylene glycol monostearate) and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 28-2 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0645] Synthesis of compound 28

[0646] Compound 28-2 (0.8 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 1.0 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 5 h, quenched with water, and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 28, 630 mg, as a white solid with an HPLC purity of 97.2%.

[0647] MS (ESI), m / z, 310.2 [ROH-OH] + .693.4[M-DAG+4Na] 4+ .ROH is stearate alcohol fragment.

[0648] Example 29: Synthesis of Compound 29

[0649] Synthesis of compound 29-2

[0650] Phospholipid compound 29-1 was synthesized using the starting material 29-SM (Synaptamide, commercially available) and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 29-2 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0651] Synthesis of compound 29

[0652] Compound 29-2 (0.53 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 1.0 mL) and acetic acid (0.1 mL) were added. The mixture was stirred at room temperature for 5 h, quenched with water, and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 29, 363 mg, as a white solid with an HPLC purity of 89.8%.

[0653] MS (ESI), m / z, 353.2 [ROH-OH] + .693.4[M-DAG+4Na] 4+ .ROH is an alkylamide alcohol fragment.

[0654] Example 30: Synthesis of Compound 30

[0655] Synthesis of compound 30-2

[0656] Phospholipid compound 30-1 was synthesized using starting material 30-SM (synthesized according to the method of compound 48 in the literature J.Med.Chem.2011,54,13,4378–4387) and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 30-2 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0657] Synthesis of compound 30

[0658] Compound 30-2 (0.45 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 1.0 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 6 h. The reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 30, 260 mg, as a light yellow solid with an HPLC purity of 93.6%.

[0659] MS (ESI), m / z, 325.2 [ROH-OH] +.693.4[M-DAG+4Na] 4+ .ROH is an amide alcohol fragment.

[0660] Example 31: Synthesis of Compound 31

[0661] Synthesis of compound 30-2

[0662] Phospholipid compound 31-1 was synthesized using starting material 31-SM (synthesized according to the method of compound 4a in the literature Chemistry–A European Journal, 4(1), 137-151.) and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 31-2 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0663] Synthesis of compound 31

[0664] Compound 31-2 (1.0 g) was dissolved in THF solution (15 mL), and TBAF (1.0 M, THF solution, 1.2 mL) and acetic acid (0.1 mL) were added. The mixture was stirred at room temperature for 6 h. The reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 31, 870 mg, as a light yellow solid with an HPLC purity of 99.0%.

[0665] MS (ESI), m / z, 455.2 [ROH-OH] + .693.4[M-DAG+4Na] 4+ .ROH is a cholesterol amide alcohol fragment.

[0666] Example 32: Synthesis of Compound 32

[0667] Synthesis of compound 32-2

[0668] Phospholipid compound 32-1 was synthesized using the starting material 32-SM (synthesized according to the method of intermediate (S)-25 in the literature Tetrahedron, 2020, 76(2):130813) and mPEG2000-SC according to the general synthesis method A of phospholipid PEG, and then compound 32-2 was synthesized with intermediate 1-5 according to the general synthesis method B of conjugated drugs.

[0669] Synthesis of compound 32

[0670] Compound 32-2 (0.8 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 1.0 mL) and acetic acid (0.1 mL) were added. The mixture was stirred at room temperature for 12 h. The reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 32, 654 mg, as a white solid with an HPLC purity of 97.7%.

[0671] MS (ESI), m / z, 522.5 [DAG-OH] + .693.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0672] Example 33: Synthesis of Compound 33

[0673] Synthesis of compound 33-2

[0674] Compound 33-2 was synthesized according to the method of compound 16 in J. Org. Chem. 1999, 64, 21, 7727-7737.

[0675] Synthesis of compound 33-3

[0676] 33-2 (0.35 g, 0.46 mmol, 1.0 eq) was dissolved in dichloromethane and triethylamine (0.24 g, 2.32 mmol, 5.0 eq), and a dichloromethane solution of mPEG-SC (1.0 g, 0.46 mmol, 1.0 eq) was added dropwise. The reaction solution was stirred at room temperature for 12 h and directly concentrated. The residue was separated and purified by column chromatography (elution with MeOH / CH2Cl2 system) to give compound 33-3, 850 mg, yield 66%.

[0677] Synthesis of compound 33-4

[0678] Compound 33-4 was synthesized according to the general synthetic method B for conjugated drugs.

[0679] Synthesis of compound 33

[0680] Compound 33-4 (0.64 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 1.0 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 6 h. The reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 33, 520 mg, as a white solid with an HPLC purity of 88.3%.

[0681] MS (ESI), m / z, 606.5 [DAG-OH] + .693.8[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0682] Example 34: Synthesis of Compound 34

[0683] Synthesis of compound 34-1

[0684] Under an ice-water bath and nitrogen protection, p-nitrophenyl chloroformate (0.53 g, 2.67 mmol, 2.0 eq) and DMAP (0.33 g, 2.67 mmol, 2.0 eq) were added to a dichloromethane solution (30 mL) of compound 33-2 (1.0 g, 1.34 mmol, 1.0 eq). The reaction solution was returned to room temperature and stirred overnight. The reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt, washed with water, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM / MeOH elution system) to obtain compound 34-1, 0.6 g, as a light yellow solid.

[0685] Synthesis of compound 34-2

[0686] Under an ice-water bath and nitrogen, mPEG2000-NH2 (1.1 g, 0.55 mmol, 1.0 eq) and DMAP (67 mg, 0.55 mmol, 1.0 eq) were added to a dichloromethane solution (20 mL) of compound 34-1 (0.5 g, 0.55 mmol, 1.0 eq). The reaction mixture was returned to room temperature and stirred for 6 h. The reaction mixture was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM / MeOH elution system) to obtain compound 34-2, 0.9 g, as a waxy solid, in a 60% yield.

[0687] Synthesis of compound 34-3

[0688] Compound 34-2 and intermediate 1-5 were used to synthesize compound 34-3 according to the general synthetic method B for conjugated drugs.

[0689] Synthesis of compound 34

[0690] Compound 34-3 (0.5 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 1.0 mL) and acetic acid (0.1 mL) were added. The mixture was stirred at room temperature for 3 h. The reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 34, 240 mg, as a white solid with an HPLC purity of 99.2%.

[0691] MS (ESI), m / z, 607.0 [DAG-OH] + .693.3[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0692] Example 35: Synthesis of Compound 35

[0693] Synthesis of compound 35-1

[0694] 1,2-Distearoyl-SN-glycero-3-phosphoethanolamine (1.0 g, 1.34 mmol, 1.0 eq) was dissolved in dimethane (20 mL) and triethylamine (0.68 g, 6.68 mmol, 5.0 eq), and a dichloromethane solution (10 mL) of mPEG1000-SC (1.53 g, 1.34 mmol, 1.0 eq) was added dropwise. The reaction solution was stirred at room temperature for 12 h. The reaction solution was directly concentrated and the residue was separated and purified by column chromatography (elution with MeOH / CH2Cl2 system) to give compound 35-1, 2.0 g, a waxy solid, with a yield of 84%.

[0695] Synthesis of compound 35-2

[0696] Compound 35-1 and intermediate 1-5 were used to synthesize compound 35-2 according to the general synthetic method B for conjugated drugs.

[0697] Synthesis of compound 35

[0698] Compound 35-2 (0.76 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 1.0 mL) and acetic acid (0.1 mL) were added. The mixture was stirred at room temperature for 12 h. The reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 35, 530 mg, as a white solid with an HPLC purity of 91.2%.

[0699] MS (ESI), m / z, 607.0 [DAG-OH] + .440.4[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0700] Example 36: Synthesis of Compound 36

[0701] Synthesis of compound 36

[0702] Compound 36 was synthesized from the starting materials 35-SM, mPEG5000-SC and intermediate 1-5 according to the synthesis method of compound 35 to obtain 870 mg of compound 36 as a white solid with an HPLC purity of 95.5%.

[0703] MS (ESI), m / z, 607.0 [DAG-OH] + .1442.0[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0704] Example 37: Synthesis of Compound 37

[0705] Synthesis of compound 37-1

[0706] At room temperature, 35-SM (200 mg, 0.27 mmol, 1.0 eq), benzoic acid (163.1 mg, 1.34 mmol, 5.0 eq), DIPEA (34 mg, 0.27 mmol, 1.0 eq), 3-methyloxazolidine-2,5-dione (Sar-NCA) (0.89 g, 7.8 mmol, 29.0 eq) and DCM (50 mL) were added in sequence. The reaction was stirred at room temperature for 1 hour. Subsequently, DIPEA (172.5 mg, 1.34 mmol, 5.0 eq) and acetic anhydride (68 mg, 0.67 mmol, 2.5 eq) were added in sequence. The reaction was stirred at room temperature for 12 hours. After completion of the reaction, the reaction solution was added dropwise to MTBE to precipitate a white precipitate. Centrifugation was performed to remove the supernatant. The solid was vacuum dried to obtain 1.1 g of a white solid product. 1 H NMR showed a degree of polymerization of about 28.

[0707] Synthesis of compound 37

[0708] Compound 37-1 was subsequently combined with intermediate 1-5 to synthesize compound 37-2 according to the general synthetic method B for conjugated drugs. Compound 37-2 (0.89 g) was dissolved in THF solution (10 mL), and TBAF (1.0 M, THF solution, 1.0 mL) and acetic acid (0.05 mL) were added. The mixture was stirred at room temperature for 12 h. The reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 37, 720 mg, as a white solid with an HPLC purity of 98.6%.

[0709] MS (ESI), m / z, 607.0 [DAG-OH] + .691.2[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0710] Example 38: Synthesis of Compound 38

[0711] Synthesis of compound 38-1

[0712] Under an ice-water bath and nitrogen protection, N-[2-(trimethylsilyl)ethoxycarbonyloxy]succinimide (0.69 g, 2.67 mmol, 2.0 eq) and DIPEA (0.52 g, 4.0 mmol, 3.0 eq) were added to a dichloromethane solution of 35-SM (1.0 g, 1.34 mmol, 1.0 eq) (20 mL). The reaction solution was then returned to room temperature and stirred for 12 h. The reaction solution was quenched with water and extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed with salt, washed with water, dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was separated and purified by silica gel column chromatography (DCM / MeOH system elution) to obtain compound 38-1, 910 mg, as a white solid, in a 77% yield.

[0713] Synthesis of compound 38-2

[0714] The raw material compound 38-1 and intermediate 1-5 were used to synthesize the compound according to the general synthesis method B of the conjugated drug.

[0715] Synthesis of compound 38-3

[0716] Compound 38-2 (1.1 g) was dissolved in THF solution (20 mL), and TBAF (1.0 M, THF solution, 2.0 mL) and acetic acid (0.2 mL) were added. The mixture was stirred at room temperature for 14 h, and the reaction solution was quenched with water and extracted with DCM (100 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography (eluted with DCM / MeOH system) to obtain compound 38-3, 830 mg, as a white solid.

[0717] Synthesis of compound 38

[0718] Oligomeric hyaluronic acid (2K-10K Da, 98.5%, 500 mg), EDC (140 mg), and NHS (85 mg) were dissolved in 5 mL of phosphate buffer (0.1 M, pH 6.2) and stirred at room temperature for 2 h. Excess EDC and NHS were removed by centrifugal ultrafiltration (MWCO 3000) and concentrated to 2 mL of aqueous solution. A DMSO solution (1 mL) of compound 38-3 (180 mg) was added and stirred at room temperature overnight. TBAF (1.0 M in THF, 2.0 mL) and acetic acid (0.2 mL) were added. The mixture was stirred at room temperature for 14 h, then directly centrifuged and ultrafiltered (MWCO 3000) and lyophilized to obtain compound 38 (450 mg) as a white solid with a GPC purity of 98.5%.

[0719] MS (ESI), m / z, 607.0 [DAG-OH] + .1004.3[M-DAG+4Na] 4+.DAG (Diglyceride) is a diglyceride fragment.

[0720] Example 39: Synthesis of Compound 39

[0721] Synthesis of compound 39-1

[0722] Oligomeric hyaluronic acid (2K-10K Da, 98.5%, 100 mg) was dissolved in water (10 mL). NH4OAc (200 mg) and sodium cyanoborohydride (20 mg x 10, added in batches) were added. The solution was heated to 40°C and reacted for 24 hours. The reaction solution was directly centrifuged and ultrafiltered (MWCO 3000) and freeze-dried to obtain 50 mg of hyaluronan-glycosylamine 39-1.

[0723] Synthesis of compound 39-2

[0724] To a THF solution (10 mL) of compound 38-3 (0.5 g, 0.4 mmol, 1.0 eq) was added succinic anhydride (200 mg, 2.0 mmol, 5.0 eq) and DIPEA (260 mg, 2.0 mmol, 5.0 eq). The reaction mixture was stirred at room temperature for 12 h. The mixture was concentrated, and the crude product was purified by silica gel column chromatography (eluting with DCM / MeOH) to afford 430 mg of compound 39-2 as a white solid.

[0725] Synthesis of compound 39

[0726] To a DMSO solution of compound 39-2 (40 mg) was added EDC (30 mg), NHS (20 mg), and DIPEA (20 mg). The mixture was stirred at room temperature for 2 h. Then, 2 mL of phosphate buffer (0.1 M, pH 6.2) containing compound 39-1 was added and the mixture was stirred at room temperature for 12 h. The reaction mixture was directly purified by centrifugal ultrafiltration (MWCO 3000) and freeze-dried to afford compound 39 (36 mg) as a white solid with a GPC purity of 95.6%.

[0727] MS (ESI), m / z, 607.0 [DAG-OH] + .1030.0[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0728] Example 40: Synthesis of Compound 40

[0729] Synthesis of compound 40-1

[0730] To a solution of fulvestrant (0.5 g, 0.83 mmol, 1.0 eq) in CH2Cl2 (10 mL) at room temperature under nitrogen was added 5-2 (0.31 g, 1.24 mmol, 1.5 eq), EDCI (0.24 g, 1.24 mmol, 1.5 eq), and DMAP (0.15 mg, 1.24 mmol, 1.5 eq) in sequence. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated and purified on a silica gel column (PE / EA as eluent) to afford 0.6 g of a solid in an 87% yield.

[0731] MS (ESI), m / z, 835.4 [M+H] + .

[0732] Synthesis of compound 40-2

[0733] To a solution of 40-1 (0.6 g) in CH2Cl2 (10 mL) was added dropwise TFA (2 mL). The reaction was stirred at room temperature for 2 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure to afford the product as a yellow solid (0.55 g) (TFA salt).

[0734] MS (ESI), m / z, 735.4 [M+H] + .

[0735] Synthesis of compound 40

[0736] At room temperature (25°C) under N2 protection, dissolve the crude product 1-6 (200 mg, 0.23 mmol, 1.0 eq) in CH2Cl2 (20 mL). Cool to 0°C, and dropwise add a solution of 40-2 (0.55 g, 0.69 mmol, 3.0 eq) in DCM (5 mL) and triethylamine (120 mg, 1.15 mmol, 5.0 eq) to the reaction mixture. After the additions are complete, remove the ice-water bath and stir at room temperature for 2 h. After completion of the reaction, concentrate under reduced pressure, dissolve in DCM (50 mL), and wash three times with water (50 mL x 3). Dry the organic phase over anhydrous Na2SO4, filter, and concentrate. Purify the crude product by column chromatography (eluting with DCM and MeOH) to yield 320 mg of the product.

[0737] MS (ESI), m / z, 607.0 [DAG-OH] + .747.1[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0738] Example 41: Synthesis of Compound 41

[0739] Synthesis of compound 41-1

[0740] To a solution of fulvestrant (0.5 g, 0.83 mmol, 1.0 eq) in CH2Cl2 (10 mL) at room temperature under nitrogen was added (tert-butoxycarbonyl)-L-valyl-L-alanine (0.36 g, 1.24 mmol, 1.5 eq), EDCI (0.24 g, 1.24 mmol, 1.5 eq), and DMAP (0.15 mg, 1.24 mmol, 1.5 eq). The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated and purified on a silica gel column (PE / EA as eluent) to afford 0.58 g of a solid in 80% yield.

[0741] MS (ESI), m / z, 877.5 [M+H] + .

[0742] Synthesis of compound 41-2

[0743] To a solution of 41-1 (0.5 g) in CH2Cl2 (10 mL) was added dropwise TFA (2 mL). The reaction was stirred at room temperature for 2 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain the product as a yellow solid (0.5 g) (TFA salt).

[0744] MS (ESI), m / z, 735.4 [M+H] + .

[0745] Synthesis of compound 41

[0746] At room temperature (25°C) under N2 protection, CH2Cl2 (20 mL) was added to the crude product 1-6 (200 mg, 0.23 mmol, 1.0 eq) to dissolve it. The temperature was lowered to 0°C, and a DCM solution (5 mL) of 41-2 (0.53 g, 0.69 mmol, 3.0 eq) and triethylamine (120 mg, 1.15 mmol, 5.0 eq) were added dropwise to the reaction solution. After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure, dissolved in DCM (50 mL), and washed three times with water (50 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by column chromatography (eluting with DCM and MeOH) to obtain 260 mg of the product with an HPLC purity of 99.2%.

[0747] MS (ESI), m / z, 607.0 [DAG-OH] + .757.6[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0748] Example 42: Synthesis of Compound 42

[0749] Synthesis of compound 42-1

[0750] At room temperature and under nitrogen protection, temozolomide 42-SM1 (0.5 g, 2.6 mmol, 1.0 eq) was added to a CH2Cl2 solution (20 mL) of temozolomide 42-SM1 (prepared according to the method of intermediate 10 in patent WO2015104684A1) (1.0 g, 3.8 mmol, 1.5 eq), HATU (1.5 g, 3.8 mmol, 1.5 eq) and DIPEA (1.0 g, 7.7 mmol, 3.0 eq). Stir at room temperature for 2 h. After the reaction is complete, the reaction solution is quenched with water and extracted with dichloromethane (50 mL x 3). The organic phases are combined, washed with salt, washed with water, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue is separated and purified by column chromatography (DCM and methanol as eluents) to obtain 0.8 g of solid with a yield of 70%.

[0751] MS (ESI), m / z, 446.1 [M+H] + .

[0752] Synthesis of compound 42-2

[0753] To a CH2Cl2 solution (5 mL) of 42-1 (0.8 g) was added diethylamine (2 mL). The reaction was stirred at room temperature for 1 h. After completion of the reaction, the residue was concentrated and purified on a reverse phase column (water and acetonitrile) and lyophilized to obtain the product 42-2, 0.3 g, in a 75% yield.

[0754] MS (ESI), m / z, 224.1 [M+H] + .

[0755] Synthesis of compound 42-3

[0756] At room temperature (25°C) under N₂ protection, to a CH₂Cl₂ solution (10 mL) of 42-2 (300 mg, 1.35 mmol, 1.0 eq) was added N-Fmoc-glycyl-glycine (0.7 g, 2.0 mmol, 1.5 eq), EDCI (0.5 g, 2.69 mmol, 2.0 eq), and DIPEA (0.52 g, 4.0 mmolm, 3.0 eq) and stirred for 2 h. The reaction solution was quenched with water and extracted with DCM (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (DCM / MeOH system) to obtain compound 42-3, 650 mg, in a yield of 86%.

[0757] MS (ESI), m / z, 560.2 [M+H] + .

[0758] Synthesis of compound 42-4

[0759] To a CH2Cl2 solution (5 mL) of 42-3 (0.6 g) was added diethylamine (2 mL). The reaction was stirred at room temperature for 1 h. After completion of the reaction, the residue was concentrated and purified on a reverse phase column (water and acetonitrile system) and lyophilized to obtain the product 42-4, 0.3 g, in an 83% yield.

[0760] MS (ESI), m / z, 338.1 [M+H] + .

[0761] Synthesis of compound 42

[0762] At room temperature (25°C) under N2 protection, CH2Cl2 (20 mL) was added to the crude product 1-6 (200 mg, 0.23 mmol, 1.0 eq) to dissolve it. The temperature was lowered to 0°C, and a DCM solution (5 mL) of 42-4 (0.23 g, 0.69 mmol, 3.0 eq) and triethylamine (120 mg, 1.15 mmol, 5.0 eq) were added dropwise to the reaction solution. After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure, dissolved in DCM (50 mL), and washed three times with water (50 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by column chromatography (eluting with DCM and MeOH) to obtain 180 mg of the product with an HPLC purity of 93.6%.

[0763] MS (ESI), m / z, 607.0 [DAG-OH] + .647.8[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0764] Example 43: Synthesis of Compound 43

[0765] Synthesis of compound 43-1

[0766] To a DMF solution (10 mL) of palbociclib (0.5 g, 1.1 mmol, 1.0 eq) was added Fmoc-Val-Ala-PAB-PNP (0.92 g, 1.34 mmol, 1.2 eq) and DIPEA (0.43 g, 3.36 mmol, 3.0 eq). The mixture was stirred at room temperature for 12 h. The reaction mixture was quenched with water and extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (DCM and methanol as eluents) to afford 0.95 g of a solid in an 86% yield.

[0767] MS (ESI), m / z, 989.5 [M+H]+ .

[0768] Synthesis of compound 43-2

[0769] To a DMF solution (10 mL) of 43-1 (0.9 g) was added diethylamine (3 mL). The reaction was stirred at room temperature for 2 h. The reaction solution was directly concentrated, and the residue was purified by reverse phase column (0.05% TFA in water and methanol) and lyophilized to obtain the product 43-2, 0.6 g, in a yield of 86%.

[0770] MS (ESI), m / z, 767.4 [M+H] + .

[0771] Synthesis of compound 43

[0772] At room temperature (25°C) under N2 protection, dissolve the crude product 1-6 (200 mg, 0.23 mmol, 1.0 eq) in CH2Cl2 (20 mL). Cool to 0°C, and dropwise add a solution of 43-2 (0.53 g, 0.69 mmol, 3.0 eq) in DCM (5 mL) and triethylamine (120 mg, 1.15 mmol, 5.0 eq) to the reaction mixture. After the additions are complete, remove the ice-water bath and stir at room temperature for 2 h. After the reaction is complete, concentrate under reduced pressure, dissolve in DCM (100 mL), and wash three times with water (50 mL x 3). The organic phase is dried over anhydrous Na2SO4, filtered, and concentrated. The crude product is purified by column chromatography (eluting with DCM and MeOH) to yield 310 mg of product with an HPLC purity of 97.2%.

[0773] MS (ESI), m / z, 607.0 [DAG-OH] + .755.1[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0774] Example 44: Synthesis of Compound 44

[0775] Synthesis of compound 44-1

[0776] To a DMF solution (10 mL) of crizotinib (0.50 g, 1.1 mmol, 1.0 eq) was added Fmoc-Val-Ala-PAB-PNP (0.92 g, 1.34 mmol, 1.2 eq) and DIPEA (0.43 g, 3.36 mmol, 3.0 eq). The mixture was stirred at room temperature for 6 h. The reaction mixture was quenched with water and extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed with salt, washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (DCM and methanol as eluents) to give 1.0 g of a solid in a 90% yield.

[0777] MS (ESI), m / z, 991.3 [M+H] + .

[0778] Synthesis of compound 44-2

[0779] To a DMF solution (10 mL) of 44-1 (1.0 g) was added diethylamine (5 mL). The reaction was stirred at room temperature for 2 h. The reaction solution was directly concentrated, and the residue was purified by reverse phase column (0.05% TFA in water and methanol) and lyophilized to obtain the product 44-2, 0.7 g, in a 90% yield.

[0780] MS (ESI), m / z, 769.2 [M+H] + .

[0781] Synthesis of compound 44

[0782] At 25°C under N2 protection, crude 1-6 (200 mg, 0.23 mmol, 1.0 eq) was dissolved in CH2Cl2 (20 mL). The temperature was lowered to 0°C, and a DCM solution (5 mL) of 44-2 (0.54 g, 0.70 mmol, 3.0 eq) and triethylamine (120 mg, 1.15 mmol, 5.0 eq) were added dropwise to the reaction mixture. After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 5 h. After the reaction was complete, the mixture was concentrated under reduced pressure, dissolved in DCM (100 mL), and washed three times with water (50 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by column chromatography (eluting with DCM and MeOH) to obtain 296 mg of product with an HPLC purity of 88.6%.

[0783] MS (ESI), m / z, 607.0 [DAG-OH] + .755.6[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0784] Example 45: Synthesis of Compound 45

[0785] Synthesis of compound 45-1

[0786] To a DMF solution (10 mL) of niraparib (0.35 g, 1.1 mmol, 1.0 eq) was added Fmoc-Val-Ala-PAB-PNP (0.92 g, 1.34 mmol, 1.2 eq) and DIPEA (0.43 g, 3.36 mmol, 3.0 eq). The mixture was stirred at room temperature for 6 h. The reaction mixture was quenched with water and extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed with salt and water, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (DCM and methanol as eluents) to obtain 0.75 g of a solid in an 80% yield.

[0787] MS (ESI), m / z, 862.4 [M+H] + .

[0788] Synthesis of compound 45-2

[0789] To a DMF solution (5 mL) of 45-1 (0.75 g) was added diethylamine (2 mL). The reaction was stirred at room temperature for 4 h. The reaction solution was directly concentrated, and the residue was purified by reverse phase column (0.05% TFA in water and methanol) and lyophilized to obtain the product 45-2, 0.5 g, in a 90% yield.

[0790] MS (ESI), m / z, 640.3 [M+H] + .

[0791] Synthesis of compound 45

[0792] At room temperature (25°C) under N2 protection, dissolve the crude product 1-6 (200 mg, 0.23 mmol, 1.0 eq) in CH2Cl2 (20 mL). Cool to 0°C, and dropwise add a solution of 45-2 (0.45 g, 0.70 mmol, 3.0 eq) in DCM (5 mL) and triethylamine (120 mg, 1.15 mmol, 5.0 eq) to the reaction mixture. After the additions are complete, remove the ice-water bath and stir at room temperature for 5 h. After completion of the reaction, concentrate under reduced pressure, dissolve in DCM (80 mL), and wash three times with water (50 mL x 3). The organic phase is dried over anhydrous Na2SO4, filtered, and concentrated. The crude product is purified by column chromatography (eluting with DCM and MeOH) to yield 160 mg of product with an HPLC purity of 99.2%.

[0793] MS (ESI), m / z, 607.0 [DAG-OH] + .723.3[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0794] Example 46: Synthesis of Compound 46

[0795] Synthesis of compound 46-1

[0796] To a solution of paclitaxel (2.0 g, 2.34 mmol, 1.0 eq) in CH2Cl2 (60 mL) at 25°C were added 5-2 (0.69 g, 2.81 mmol, 1.2 eq), DMAP (0.86 g, 7.03 mmol, 3.0 eq), and EDCI (1.34 g, 7.03 mmol, 3.0 eq) in sequence. The mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction solution was poured into saturated aqueous ammonium chloride and washed thoroughly (150 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, dried in vacuo, and purified on a silica gel column (PE / EA = 1:2 to 1:3 as eluent) to obtain 2.0 g of the pure product in a yield of 79%.

[0797] MS (ESI), m / z, 982.4 [M-100] + .

[0798] Synthesis of compound 46-2

[0799] A formic acid solution (10 mL) of compound 46-1 (2.0 g, 1.85 mmol, 1.0 eq) was stirred at 0°C for 6 h. After completion, the reaction solution was concentrated to 3 mL at 0°C and tert-butyl methyl ether (100 mL) was added dropwise to precipitate a large amount of white solid. The filter cake was filtered and thoroughly dried under vacuum to yield 1.5 g of a pure white solid.

[0800] MS (ESI), m / z, 982.4 [M+H] + .

[0801] Synthesis of compound 46

[0802] At room temperature (25°C) under N2 protection, dissolve the crude product 1-6 (755 mg, 0.27 mmol, 1.0 eq) in CH2Cl2 (10 mL). Cool to 0°C, and dropwise add a solution of 46-2 (1.36 g, 1.38 mmol, 5.0 eq) in DCM (2 mL) and triethylamine (271 mg, 2.69 mmol, 10 eq) to the reaction mixture. After the addition is complete, remove the ice-water bath and stir at room temperature for 2 h. TLC confirms the reaction is complete. The reaction mixture is concentrated under reduced pressure, dissolved in DCM (100 mL), and then washed three times with saturated brine (150 mL x 3). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude product was passed through a normal phase column, first using an EA / MeOH = 1:0-10:1 elution system, and then switched to an elution system DCM / MeOH = 95:5-93:7-92:8-89:11 to obtain 650 mg of the product.

[0803] MS (ESI), m / z, 607.0 [DAG-OH] + .808.8[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0804] Example 47: Synthesis of Compound 47

[0805] Synthesis of compound 47-1

[0806] At room temperature (25°C) and under nitrogen protection, DIPEA (1.14 g, 8.82 mmol, 3.0 eq) was added to a solution of octreotide (3.0 g, 2.94 mmol, 1.0 eq) in DMF (60 mL). The reaction solution was cooled to -40°C, and a solution of tert-butyl N-succinimidyl carbonate (633 mg, 2.94 mmol, 1.0 eq) in DMF (10 mL) was slowly added dropwise. The reaction mixture was stirred at -40°C for 0.5 hours, then naturally warmed to room temperature and stirred for another 2 hours. The reaction was completed by LCMS. The reaction solution was concentrated and purified on a silica gel column (eluted with DCM / MeOH system) to obtain product 47-1 as a white solid, 2.8 g, in 85% yield.

[0807] MS (ESI), m / z, 1118.40 [M+H] + .

[0808] Synthesis of compound 47-2

[0809] To a DMF solution (10 mL) of 47-1 (1.5 g, 1.34 mmol, 1.0 eq) were added Fmoc-Val-Ala-PAB-PNP (0.9 g, 1.34 mmol, 1.0 eq) and DIPEA (0.43 g, 3.36 mmol, 3.0 eq). The mixture was stirred at room temperature for 4 h. The reaction mixture was quenched with water and concentrated. The residue was directly purified by reverse-phase column chromatography (eluted with 0.05% TFA, H₂O, and methanol). The product peak was collected, concentrated, and lyophilized to obtain the solid product 47-2 (1.3 g, 58% yield).

[0810] MS (ESI), m / z, 1660.5 [M+H] + .

[0811] Synthesis of compound 47-3

[0812] To a DMF solution (10 mL) of 47-2 (1.2 g) was added diethylamine (3 mL). The reaction was stirred at room temperature for 2 h. The reaction solution was directly concentrated, and the residue was purified on a reverse phase column (0.05% TFA in water and methanol). The product peak was collected, concentrated, and lyophilized to obtain a solid product 47-3, 0.84 g, in an 81% yield.

[0813] MS (ESI), m / z, 1438.6 [M+H] + .

[0814] Synthesis of compound 47-4

[0815] At room temperature (25°C) under N2 protection, CH2Cl2 (10 mL) was added to the crude product 1-6 (300 mg, 0.11 mmol, 1.0 eq) to dissolve it. The temperature was lowered to 0°C, and a DCM / DMF solution (2 / 0.5 mL) of 47-3 (0.77 g, 0.54 mmol, 5.0 eq) and triethylamine (108 mg, 1.07 mmol, 10.0 eq) were added dropwise to the reaction solution. After the addition was complete, the ice-water bath was removed and the product was stirred at room temperature for 12 h. The reaction solution was directly concentrated, and the crude product was first purified by normal phase column chromatography (DCM / MeOH system) and then by reverse phase column chromatography (0.05% TFA in water and methanol system). The product peak was collected, concentrated, and lyophilized to obtain a solid product 47-4 (180 mg).

[0816] MS (ESI), m / z, 607.0 [DAG-OH] + .923.0[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0817] Synthesis of compound 47

[0818] To a solution of 47-4 (100 mg) in CH2Cl2 (5 mL) was added trifluoroacetic acid (1 mL) in an ice-water bath at 5°C under N2 protection. The reaction solution was warmed to room temperature and stirred for 1 h. The reaction was monitored by HPLC for completion. The product was concentrated at low temperature (5°C), dissolved in DMF (1 mL), and purified by Prep-HPLC (0.05% TFA in water and acetonitrile system). The product peak was collected and lyophilized to obtain a solid product 47, 54 mg.

[0819] MS (ESI), m / z, 607.0 [DAG-OH] + .897.8[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0820] Example 48: Synthesis of Compound 48

[0821] Synthesis of compound 48-1

[0822] Oxaliplatin (2.0 g, 5.03 mmol) was added to water (50 mL), followed by 30% hydrogen peroxide (50 mL). The mixture was heated at 50°C for 1 h. The reaction solution was concentrated, and 50 mL of ethanol was added to precipitate a solid, which was filtered and dried under vacuum to afford compound 48-1 as a white solid (2.0 g, 92% yield).

[0823] MS (ESI), m / z, 395.8 [M+H-2H2O] + .

[0824] Synthesis of compound 48-2

[0825] 48-1 (1.0 g, 2.32 mmol, 1.0 eq) and Boc-Gly-Gly-Osu (0.76 g, 2.32 mmol, 1.0 eq) were added to DMSO (10 mL), and the mixture was heated to 60°C for 12 h. The reaction solution was added dropwise to 100 mL of glacial MTBE, and the solid was collected by centrifugation and dried in vacuo to obtain the crude product compound 48-2, 1.2 g, with a yield of 80%.

[0826] MS (ESI), m / z, 590.2 [M-55] + .

[0827] Synthesis of compound 48-3

[0828] Trifluoroacetic acid (5 mL) was added to a DCM solution (10 mL) of compound 48-2 (1.0 g). The reaction solution was stirred at room temperature for 1 h, concentrated, washed twice with MTBE, and the solid was dried in vacuo to obtain compound 48-3 (TFA salt), 0.7 g, yield 78%.

[0829] MS (ESI), m / z, 546.2 [M+H] + .

[0830] Synthesis of compound 48

[0831] At room temperature (25°C) and under N2 protection, CH2Cl2 (10 mL) was added to the crude product 1-6 (500 mg, 0.18 mmol, 1.0 eq) to dissolve it. The temperature was lowered to 0°C, and a DCM solution (2 mL) of 48-3 (0.49 g, 0.89 mmol, 5.0 eq) and triethylamine (180 mg, 1.78 mmol, 10.0 eq) were added dropwise to the reaction solution. After the addition was complete, the ice-water bath was removed and the mixture was stirred at room temperature for 6 h. The reaction solution was concentrated to 5 mL and added dropwise to a MTBE solution (100 mL). After centrifugation, the solid was collected to obtain 460 mg of the crude product, which was further purified by Sephadex LH-20 column.

[0832] MS (ESI), m / z, 607.0 [DAG-OH] + .699.8[M-DAG+4Na] 4+ .DAG (Diglyceride) is a diglyceride fragment.

[0833] Evaluation Example 1: Drug Release Study

[0834] PBS release studies:

[0835] Experimental Method: At room temperature, weigh 2.0 mg of phospholipid conjugate and dissolve it in 20 mL of distilled water. Use a pipette to transfer 1.0 mL of the solution to a 50 mL volumetric flask and dilute to 50 mL with PBS buffer (pH 5.0). Place the solution in a 37°C water bath in a constant-temperature oscillator. Sample 0.5 mL at regular intervals for HPLC analysis of drug release.

[0836] Experimental results:

[0837] Table 1 Drug release test of the compounds of the present invention in PBS

[0838] Protease release studies:

[0839] Papain activation: Dissolve 10 mg of papain in 1.0 mL of 10 mM PBS (pH 5.0), add 1.5 mL of 30 mM DTT / 15 mM EDTA solution, shake at 37°C for 30 minutes, and cool to room temperature.

[0840] At room temperature, 4 mg of phospholipid conjugates of Examples 5, 9, 15, and 16 were accurately weighed and dissolved in 10 mM sodium acetate buffer 5.0 (1.0 mL). 0.5 mL of each was added to 0.1 mL of the activated papain solution described above. The mixture was shaken in a 37°C water bath for 6 h and 24 h, respectively. At the time points, 1.4 mL of 0.05% TFA acetonitrile was added, the volume was adjusted to 2.0 mL, filtered, and analyzed by HPLC. The experimental results showed that under these conditions, the SN38 release rates of compound 5 were 25% and 65% after 6 h and 24 h, respectively. Under these conditions, the SN38 release rates of compound 9 were 45% and 95% after 6 h and 24 h, respectively. The release rates of compounds 15 and 16 were <1%.

[0841] DTT Release Studies:

[0842] At room temperature, 2 mg of each of the phospholipid conjugates of Examples 10, 15, and 16 were accurately weighed and dissolved in 10 mM phosphate buffer 7.0 (0.5 mL). 0.5 mL of 20 mM DTT solution was added (final DTT concentration was 10 mM). The mixture was shaken in a 37°C water bath for 6 h. 1.4 mL of 0.05% TFA acetonitrile was added, the volume was adjusted to 2.0 mL, and the mixture was filtered and analyzed by HPLC. The experimental results showed that under these conditions, the SN38 release rate of compound 10 was 100%, while the release rates of compounds 15 and 16 were <1%.

[0843] The results in Table 1 show that Example compounds 1, 5, and 7 containing responsive linkers can release the drug SN38 under PBS pH = 5.0 and 37°C, while compounds 15 and 16 without responsive linkers hardly release the drug under these conditions. Furthermore, protease-responsive compound 9 and reduction-responsive compound 10 can both release the drug SN38 under their respective responsive release conditions, while compounds 15 and 16 without responsive linkers also do not release the drug.

[0844] Evaluation Example 2: Particle Size Test of Example 1

[0845] Sample preparation: Weigh about 10 mg of the compound of Example 1, add 1 mL of the above-mentioned distilled water to dissolve, filter through a 0.22 μm aqueous filter membrane, and set aside.

[0846] Sample testing: Slowly transfer the sample solution to a cuvette using a dropper for testing. The testing instrument is a Zeta sizer Pro Blue nanoparticle sizer.

[0847] Test results: The nanoparticle size data of the compound of the present invention are shown in Figure 1 and Table 2.

[0848] Table 2 Nanoparticle size data of the compounds of the present invention

[0849] The results in Table 2 show that the average particle size of the compounds of the present invention is in the range of 10 - 200 nM, and the nano size varies according to different drugs and linking methods.

[0850] Evaluation Example 3: Pharmacodynamic study of Example 1 on a human hepatocellular carcinoma HepG2 xenograft mouse model

[0851] Experimental animals and inoculation method:

[0852] BALB / c nude mice, female, 4 - 5 weeks old, weighing 20 - 25 grams. After arrival, they were housed in the experimental environment for one week. HepG2 tumor blocks were subcutaneously inoculated on the right back of the nude mice, and grouping and dosing started when the tumor grew to ~170 mm 3 , with 6 mice in each group.

[0853] Preparation of test samples:

[0854] Weigh an appropriate amount of the compound, add a certain volume of normal saline, shake to dissolve, and filter through a 0.2 μM filter membrane for standby.

[0855] Administration of test drugs:

[0856] The dosing dose and dosing regimen are shown in Table 2. The tumor volume under the skin of the nude mice was measured 2 - 3 times a week (the formula for calculating the tumor volume is: V = 0.5a × b 2 , where a and b represent the major and minor axes of the tumor respectively), the mouse weight was measured, and the data were recorded.

[0857] Table 3 Note: The dosing volume is 10 mg / mL.

[0858] Analysis and evaluation:

[0859] Experimental evaluation index: Evaluation was carried out using the tumor growth inhibition rate TGI (%) or the relative tumor proliferation rate T / C (%). Among them, T is the experimental group and C is the control group.

[0860] Calculation of the relative tumor proliferation rate T / C (%): If T > T0, T / C (%) = (T - T0) / (C - C0) × 100%; if T < T0, T / C (%) = (T - T0) / T0 × 100%. Here, T and C are the tumor volumes at the end of the experiment; T0 and C0 are the tumor volumes at the start of the experiment.

[0861] Calculation of the tumor growth inhibition rate TGI (%): TGI (%) = (1 - T / C) × 100%.

[0862] Evaluation criteria: T / C (%)>40 (i.e. TGI (%) <60%) is invalid; T / C (%)≤40 (i.e. TGI (%) ≥60%) is effective, and after statistical processing, P<0.05 is effective.

[0863] Results of efficacy experiments:

[0864] The tumor inhibitory effect of Example 1 on the human liver cancer HepG2 tumor model is shown in FIG2 .

[0865] The results showed that Example 1 of the present invention had a very strong inhibitory effect on the tumor growth of HepG2 cell nude mouse model, with a tumor inhibition rate of 99%, and the tumor almost completely disappeared.

[0866] Evaluation Example 4: Pharmacodynamic Study of Examples 5, 7, 15 and 16 on Human Hepatocellular Carcinoma HepG2 Xenograft Mouse Model Experimental Animals and Inoculation Methods:

[0867] Female BALB / c nude mice, 4-5 weeks old, weighing 20-25 g, were housed in the experimental environment for one week upon arrival. HepG2 tumors were subcutaneously inoculated on the right back of the nude mice. The tumors grew to ~170 mm. 3 The mice were divided into groups and dosed at 4 ℃ and 8 ℃, with 6 mice in each group.

[0868] Preparation of test samples:

[0869] Weigh an appropriate amount of compound, add a certain volume of physiological saline, shake to dissolve, filter with a 0.2 μM filter membrane, and set aside.

[0870] Administration of test drugs:

[0871] The dosage and administration schedule are shown in Table 3. The subcutaneous tumor volume of nude mice was measured 2-3 times a week (the calculation formula of tumor volume is: V = 0.5a × b 2 , a and b represent the long diameter and short diameter of the tumor, respectively), weigh the mice, and record the data.

[0872] Table 4 Note: The dosage volume is 10 mg / mL.

[0873] Analysis and evaluation:

[0874] The analysis and evaluation indexes, calculation of the relative tumor proliferation rate T / C (%), and calculation and evaluation criteria of the tumor growth inhibition rate TGI (%) were the same as those in Example 3.

[0875] Results of efficacy experiments:

[0876] The tumor inhibitory effects of Examples 5, 7, 15 and 16 on the human liver cancer HepG2 tumor model are shown in FIG3 .

[0877] The results showed that Example compounds 5 and 7 containing environmentally sensitive releasable linkers had a very strong inhibitory effect on tumor growth in the HepG2 cell nude mouse model. Among them, Examples 5 and 7 could completely regress the tumor, with tumor inhibition rates reaching 99%. Compounds 15 and 16 that did not contain sensitive linkers had very poor tumor inhibition effects (tumor inhibition rates were 0% and 6%, respectively). It can be seen that whether or not a compound contains a sensitive linker has a great influence on its efficacy.

[0878] Evaluation Example 5: Pharmacodynamic Study of Examples 1, 5, and 7 in Human Gastric Cancer Cell NCI-N87 Xenograft Mouse Model

[0879] Experimental animals and inoculation methods:

[0880] Female BALB / c nude mice, 4-5 weeks old, weighing 20-25 g, were housed in the experimental environment for one week upon arrival. NCI-N87 cancer cells were subcutaneously inoculated on the right back of the nude mice, and tumors grew to ~180 mm. 3 The mice were divided into groups and dosed at 4 ℃ and 8 ℃, with 6 mice in each group.

[0881] Preparation of test samples:

[0882] Weigh an appropriate amount of compound, add a certain volume of physiological saline, shake to dissolve, filter with a 0.2 μM filter membrane, and set aside.

[0883] Administration of test drugs:

[0884] The dosage and administration schedule are shown in Table 2. The subcutaneous tumor volume of nude mice was measured 2-3 times a week (the calculation formula of tumor volume is: V = 0.5a × b 2 , a and b represent the long diameter and short diameter of the tumor, respectively), weigh the mice, and record the data.

[0885] Table 5 Note: The dosage volume is 10 mg / mL.

[0886] Analysis and evaluation:

[0887] The analysis and evaluation indexes, calculation of the relative tumor proliferation rate T / C (%), and calculation and evaluation criteria of the tumor growth inhibition rate TGI (%) were the same as those in Example 3.

[0888] Results of efficacy experiments:

[0889] The tumor inhibitory effects of Examples 1, 5 and 7 on the human gastric cancer NCI-N87 tumor model are shown in FIG4 .

[0890] The changes in animal body weight during the efficacy evaluation of Examples 1, 5 and 7 are shown in FIG5 .

[0891] The results demonstrated that compounds of Examples 1, 5, and 7 of the present invention exhibited highly potent inhibitory effects on the human gastric cancer NCI-N87 tumor model, with the 10 mg / kg dose of Examples 5 and 7 achieving tumor volume inhibition rates of 86% and 77%, respectively. At the end of the experiment, the weight gain of the animals in each group during the dosing period was not significantly different from that in the control group.

[0892] Evaluation Example 6: Pharmacodynamic Study of Example 5 and T-Dxd in Human Colon Cancer Cell HT-29 Xenograft Mouse Model

[0893] Experimental animals and inoculation methods:

[0894] Female BALB / c nude mice, 4-5 weeks old, weighing 20-25 g, were housed in the experimental environment for one week upon arrival. HT-29 cancer cells were subcutaneously inoculated on the right back of the nude mice, and the tumors grew to ~180 mm. 3 The mice were divided into groups and dosed at 4 ℃ and 8 ℃, with 6 mice in each group.

[0895] Preparation of test samples:

[0896] Weigh an appropriate amount of the compound of this example, add a certain volume of physiological saline, shake to dissolve, filter through a 0.2 μM filter membrane, and set aside.

[0897] Administration of test drugs:

[0898] The dosage and administration schedule are shown in Table 2. The subcutaneous tumor volume of nude mice was measured 2-3 times a week (the calculation formula of tumor volume is: V = 0.5a × b 2 , a and b represent the long diameter and short diameter of the tumor, respectively), weigh the mice, and record the data.

[0899] Table 6 Note: The dosage volume is 10 mg / mL.

[0900] Analysis and evaluation:

[0901] The analysis and evaluation indexes, calculation of the relative tumor proliferation rate T / C (%), and calculation and evaluation criteria of the tumor growth inhibition rate TGI (%) were the same as those in Example 3.

[0902] Results of efficacy experiments:

[0903] The tumor inhibitory effects of Example 5 and T-Dxd on the human colon cancer cell HT-29 tumor model are shown in FIG6 .

[0904] The results showed that the inhibitory effect of the compound Example 5 on human colon cancer cell HT-29 was significantly better than that of the ADC drug T-Dxd, with the tumor inhibition rates of the 10 mg / kg dose group being 77% and 49%, respectively.

[0905] Evaluation Example 7: Pharmacodynamic Study of Example 5 in Human Fibrosarcoma Cell HT-1080 Xenograft Mouse Model

[0906] Experimental animals and inoculation methods:

[0907] Female BALB / c nude mice, 4-5 weeks old, weighing 20-25 g, were housed in the experimental environment for one week upon arrival. HT-1080 cancer cells were subcutaneously inoculated on the right back of the nude mice until the tumors grew to ~400 mm. 3 The mice were divided into groups and dosed at 4 ℃ and 8 ℃, with 6 mice in each group.

[0908] Preparation of test samples:

[0909] Weigh an appropriate amount of the compound of this example, add a certain volume of physiological saline, shake to dissolve, filter through a 0.2 μM filter membrane, and set aside.

[0910] Administration of test drugs:

[0911] The dosage and administration schedule are shown in Table 2. The subcutaneous tumor volume of nude mice was measured 2-3 times a week (the calculation formula of tumor volume is: V = 0.5a × b 2 , a and b represent the long diameter and short diameter of the tumor, respectively), weigh the mice, and record the data.

[0912] Table 7 Note: The dosage volume is 10 mg / mL.

[0913] Analysis and evaluation:

[0914] The analysis and evaluation indexes, calculation of the relative tumor proliferation rate T / C (%), and calculation and evaluation criteria of the tumor growth inhibition rate TGI (%) were the same as those in Example 3.

[0915] Results of efficacy experiments:

[0916] The tumor inhibitory effect of Example 5 on the human fibrosarcoma cell HT-1080 tumor model is shown in FIG7 .

[0917] The results showed that the compound of Example 5 of the present invention had a significant tumor-inhibiting effect on human fibrosarcoma cells HT-1080.

[0918] Evaluation Example 8: Pharmacodynamic Study of Examples 5 and 46 in an Orthotopic U87-MG Glioma Mouse Model

[0919] Experimental animals and inoculation methods:

[0920] BALB / c nude mice, female, 6-8 weeks old, weighing 18-20 g, were housed in the experimental environment for one week after arrival. 72 mL of U87MG-Luc2 cell suspension at 100 cells / mL was pipetted until uniform and placed on ice. After removing the hair at the inoculation site, the mouse inoculation site was disinfected with iodine cotton balls. The animal was fixed on a mouse brain stereotaxic instrument and 2 μL of cell suspension was inoculated into the mouse brain with a microsyringe. 5×10 4 On day 8 after inoculation, animals were weighed and live imaging was performed. Based on body weight and fluorescence imaging data, U87MG-Luc2 model animals were randomly divided into groups of 8. The day of grouping was considered experimental day 0, and drug administration was performed after grouping.

[0921] Preparation of test samples:

[0922] Weigh an appropriate amount of the compound of the present invention, add a certain volume of physiological saline, shake to dissolve, filter through a 0.2 μM filter membrane, and set aside.

[0923] Administration of test drugs:

[0924] The dosage and administration schedule are shown in Table 8. The subcutaneous tumor volume of nude mice was measured 2-3 times a week (the calculation formula of tumor volume is: V = 0.5a × b 2 , a and b represent the long diameter and short diameter of the tumor, respectively), weigh the mice, and record the data.

[0925] Table 8 Note: The dosage volume is 10 mg / mL.

[0926] Analysis and evaluation:

[0927] Measurement Indicators: Mice were weighed twice weekly after the start of dosing. In vivo imaging and quantitative analysis (TGI%) of the fluorescent cell line model were performed twice weekly. Prior to in vivo imaging, animals were intraperitoneally injected with D-luciferin at a concentration of 15 mg / mL in a 0.2 mL volume. In vivo imaging was performed 10 minutes after D-luciferin injection.

[0928] Animals were examined daily and the time of death was recorded. If an animal's health deteriorated or became moribund (animals showed significant weight loss, with a weight loss greater than 20%), could not eat or drink normally, or had difficulty moving or became paralyzed, the animal was considered dead and immediately euthanized with CO2. The mean survival time (MST) of each group was calculated.

[0929] Results of efficacy experiments:

[0930] The tumor inhibitory effects and survival curves of Example 5 and Example 46 on the U87-MG brain glioma orthotopic mouse model are shown in FIG8 .

[0931] The results showed that Examples 5 and 46 of the present invention had very strong tumor inhibitory effects in the U87-MG brain glioma orthotopic mouse model, with significant differences compared to the blank control group, and the survival periods were increased by 15 days and 30 days, respectively. This indicates that the conjugated drug Examples 5 and 46 of the present invention can break through the blood-brain barrier, carry drugs and accumulate in brain tumor tissue, and have potential application value in the field of brain tumor treatment.

[0932] Although the above describes specific embodiments of the present invention, it should be understood by those skilled in the art that these are merely illustrative and that various changes or modifications may be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the scope of protection of the present invention is defined by the appended claims.

Claims

1. A phospholipid drug conjugate as shown in formula (I), or a stereoisomer of the phospholipid drug conjugate, a prodrug thereof, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof, in: X 0 O or S; X 1 O, S or NR 1a ; X 2 O, S or NR 2a ; L 1 , L 2 for non-existent or connected units; M 0 OR 0a , SR 0b NR 0c R 0d or T 0 -(LD) n ; K 1 P 1 or Y 1 is the branching center, P 1 M is the pharmacokinetic regulation unit. 1 T 1 -(LD) n ; Y 2 It is the branching center; M 2 Does not exist or T 2 -(LD) n ; M 0 、M 1 and M 2 At least one of them contains an LD fragment; Q 2a T 2a -R 1 Or not exist, Q 2b T 2b -R 2 Or not exist, Q 2a and Q 2b At least one exists; T 0 , T 1 , T 2 Each independently selected from absence or extension; T 2a 、T 2b for non-existent or connected units; L is L 3 -L s , L 3 is an environmental responsive linker, wherein the environmental responsive linker is selected from an enzyme responsive linker, a pH responsive linker, a light responsive linker, and a redox responsive linker; L s It is present or absent, and when present, it is a self-releasing connecting part; D is a bioactive molecular fragment. When the conjugate contains multiple Ds, Ds may be the same or different. R 1a and R 2a Each independently selected from hydrogen, C1-C 10 Alkyl, C1~C 10 Alkoxy, C3~C 10 Alkenyl, C3~C 10 Alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C 10 Aryl, C5~C 10 Heteroaryl or amino protecting groups; R 0a Each independently selected from hydrogen, C1-C 10 Alkyl, C1~C 10 Alkoxy, C3~C 10 Alkenyl, C3~C 10 Alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C 10 Aryl, C5~C 10 Heteroaryl or hydroxy protecting groups; R 0b Selected from hydrogen, C1~C 10 Alkyl, C1~C 10 Alkoxy, C3~C 10 Alkenyl, C3~C 10 Alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C 10 Aryl, C5~C 10 Heteroaryl or thiol protecting groups; R 0c and R 0d Each independently selected from hydrogen, C1-C 10 Alkyl, C1~C 10 Alkoxy, C3~C 10 Alkenyl, C3~C 10 Alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C 10 Aryl, C5~C 10 Heteroaryl or amino protecting groups; R 1 and R 2 Each independently selected from saturated or unsaturated C1-C 50 Alkyl, cholesterol, retinoic acid and tocopherol, the C1~C 50 Alkyl, cholesterol, retinoic acid and tocopherol are optionally substituted by one or more selected from hydroxyl, halogen, C 1-10 Alkyl, C 1-10 alkyl Oxygen, C 3-12 The substituents of cycloalkyl and 3-12 membered heterocycloalkyl are substituted, the C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 Cycloalkyl and 3-12 membered heterocycloalkyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-6 Substitution of cycloalkyl groups n is an integer from 1 to 10. When n>1, LD may be the same or different; One or more of the hydrogen atoms connected to carbon atoms in the compound represented by formula (I) may be replaced by deuterium; In the compound represented by formula (I), one or more hydrogen atoms connected to carbon atoms may be replaced by any substituent; Phospholipid isomers include compounds with phosphine as chiral center, selected from: or a chiral mixture thereof.

2. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claim 1, characterized in that: The phospholipid core in formula (I) Meet one or more of the following conditions: (1) (2) (3) Where X 2 , X 1 、M 0 At least one atom connected to P is not an oxygen atom.

3. The phospholipid drug conjugate according to claim 1-2, or its stereoisomers, prodrugs, pharmaceutically acceptable salts or pharmaceutically acceptable solvates, characterized in that: The phospholipid drug conjugate has a nanometer size; preferably, the nanometer size is an average nanometer particle size range of 1-500 nanometers; more preferably, the average particle size range is 5-100 nanometers.

4. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claims 1-3, characterized in that: Meet one or more of the following conditions: (1)L 1 , L 2 is absent or present. When present, L 1 , L 2 Each is independently an alkane, a heterocycloalkane, an aromatic hydrocarbon, a heterocycloalkane, a substituted or unsubstituted amino acid or a polypeptide composed thereof, or a combination thereof, wherein L 1 , L 2 The functional group connecting parts at both ends are selected from: Among them, R La , R Lb , R Lc , R Ld Each independently selected from hydroxyl, C1-C5 alkyl or C1-C5 alkoxy; R Le is hydrogen, hydroxy, C1-C5 alkyl or C1-C5 alkoxy; L 1a , L 1b Each independently selected from O, NR Lf , S, SS, Or directly connected (not present), R Lf is hydrogen or C1-C5 alkyl; (2)T 0 、T 1 、T 2 Each is independent of being absent or present. When present, T 0 、T 1 、T 2 Each is independently an alkane, a heterocycloalkane, an aromatic hydrocarbon, a heterocycloalkane, a substituted or unsubstituted amino acid or a polypeptide composed thereof, or a combination thereof, wherein T 0 、T 1 、T 2 The functional groups at the two ends of each connection are selected from: Among them, R La , R Lb , R Lc , R Ld Each independently selected from hydroxyl, C1-C5 alkyl or C1-C5 alkoxy; R Le is hydrogen, hydroxy, C1-C5 alkyl or C1-C5 alkoxy; L 1a , L 1b Each independently selected from O, NR Lf , S, SS, Or directly connected (not present), R Lf is hydrogen or C1-C5 alkyl; (3)T 2a , T 2b is absent or present. When present, T 2a , T 2b Each is independently an alkane, a heterocycloalkane, an aromatic hydrocarbon, a heterocycloalkane, a substituted or unsubstituted amino acid or a polypeptide composed thereof, or a combination thereof, wherein T 2a , T 2b Connection points at both ends The functional group is selected from: Among them, L 1a , L 1b Each independently selected from O, NR Lf , S, SS, Or directly connected (not present), R Lf is hydrogen or C1-C5 alkyl; (4)L 3 is selected from enzyme-responsive linkers, pH-responsive linkers, light-responsive linkers and redox-responsive linkers, preferably selected from enzyme-responsive linkers, pH-responsive linkers and redox-responsive linkers, more preferably enzyme-responsive linkers; (5)Y 1 , Y 2 is at least a trifunctional branching center selected from a trifunctional amino acid, a polypeptide or the following structures: Among them, R y0 A group selected from hydrogen, deuterium, halogen, nitro, cyano, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl, or a group containing a primary amine, secondary amine, tertiary amine, hydroxyl, sulfhydryl, carboxyl, ester, amide, boric acid, boric ester, phosphoric acid, sulfonic acid, sulfoxide, aldehyde, or keto functional group; R y1 Selected from C1-C10 alkyl, C1-C10 alkoxy, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl; R y2 A group selected from hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl, or a group containing a primary amine, secondary amine, tertiary amine, hydroxyl, sulfhydryl, carboxyl, ester, amide, boric acid, boric ester, phosphoric acid, sulfonic acid, sulfoxide, aldehyde, or keto functional group; Ring A is C6~C 20 Aryl or C5~C 20 Heteroaryl; the C5~C 20 The heteroatom in the heteroaryl group is O, S or N, and the number of the heteroatoms is one or more. When there are multiple heteroatoms, the heteroatoms are the same or different; The E ring is a C2-C8 heterocycloalkyl group; the heteroatom in the heterocycloalkyl group is O, S or N, and the number of the heteroatoms is one or more. When there are multiple heteroatoms, the heteroatoms are the same or different; (6)P 1 The pharmacokinetic regulating unit is selected from polyethylene glycol, sugar, oligosaccharide and polysaccharide (such as hyaluronic acid, cartilage sulfate The invention can be used in combination with any of the following: oligosaccharides, ...

5. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claims 1-4, characterized in that: Meet one or more of the following conditions: (1) When the L 3 It is an enzyme-responsive linker that can be cut by one or more of the following enzymes: secretory phospholipase A2, acid phosphatase, serum alkaline phosphatase, cytochrome P450, sulfatase, prostate-specific antigen, phospholipase A1, phospholipase A2, phospholipase B, phospholipase C, phospholipase D, neutrophil elastase, cysteine ​​protease-3, cathepsin, matrix metalloproteinase, β-glucuronidase, β-galactosidase, nitroreductase, reduced coenzyme II, aminopeptidase N, carboxylesterase, diaphorase, histone deacetylase, asparagine endopeptidase, urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor, collagenase; preferably, it is cut by one or more of the following enzymes: cysteine ​​protease-3, cathepsin, matrix metalloproteinase, elastase or β-glucuronidase; (2) When the L 3 is a pH-responsive linker comprising the following structures: one or more of a hydrazone, an imine, an oxime, a carboxylate, a thioester, a sulfate, a sulfonate, an orthoester, a carbonate, a carbamate, a substituted carbamate, a ketal, an acetal, a silyl ether, a phosphate, a borate, a phosphoramide or a cis-aconitic acid group; (3) When the L 3 is a photoresponsive linker comprising the following structures: one or more of o-nitrobenzene, coumarin, benzoin, BODIPY or cyanine groups; (4) When the L 3 It is a redox-responsive linker, which contains the following structures: one or more of thioketal, phenylboronic acid, phenylboronic acid, oxalate, vinyl ether, thioether, aminoacrylate, disulfide, diselenyl, 2,4-dinitrobenzenesulfonate, 2-azidomethylbenzoate, 4-azidobenzyl, unsaturated acid ester or azobenzene group.

6. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claims 1-5, characterized in that: Meet one or more of the following conditions: (1) When L 3 is an enzyme-responsive linker selected from a short peptide consisting of 2-10 amino acid residues Val, D-Val, Cit, Phe, Lys, Leu, Gly, Ala, Asn, Asp, Arg, Pro, Tyr, Ile, His, Ser, Gln, Glu, Met, preferably a short peptide consisting of 2-10 amino acids selected from Val, Cit, Phe, Lys, Ala, Leu, Gly, Asn, Asp; preferably, L 3 Selected from the following structures: (2) When L s When present, it is selected from the following structures: Preferably, L s Selected from the following structures: (3) When L 3 is a pH-responsive linker selected from the following structures: Among them, pm = 0 ~ 4; R p3 , R q1 Selected from H, substituted or unsubstituted C1-C10 alkyl; Preferably, L 3 Select from the following structures: (4) When the L 3 is a light-responsive linker selected from the following structures: (5) When L 3 It is a redox-responsive linker, which comprises the following structure:

7. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claims 1-4, characterized in that: P1 is selected from: (1) The polyethylene glycol derivative residue is selected from the following structures: wherein pa1 is selected from an integer between 5 and 250, pr1 is selected from an integer between 0 and 8, and R p1 is hydrogen, C1-C10 alkyl, C1-C10 heteroalkyl, C3-C10 cycloalkyl, C3-C10 alkenyl, C3-C10 alkynyl or a hydroxy protecting group; Preferably, pa1 is selected from an integer between 5 and 150, pr1 is selected from an integer between 0 and 8, and R p1 is hydrogen, C1-C10 alkyl or hydroxyl protecting group; More preferably, pa1 is selected from an integer between 10 and 60, pr1 is selected from an integer between 1 and 2, and R p1 is a C1-C10 alkyl group; (2) The polycreatine derivative residue is selected from the following structures: Wherein, pb1 is selected from an integer between 5 and 250, R p2 R is selected from hydrogen, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl or an amine protecting group; p3 Selected from OR p3a or NR p3b R p3c , where R p3a is selected from hydrogen, C1-C10 alkyl, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl or C5-C10 heteroaryl; R p3b , R p3c Each is independently selected from hydrogen, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl, amino protecting group or R p3b and R p3c The nitrogen atom they are connected to together forms a C3-C10 ring structure; Preferably, pb1 is selected from an integer between 5 and 150; (3) The quaternary ammonium salt derivative residue is selected from the following structures: Among them, pc1 is selected from integers between 0 and 10, A - Selected from R p3 , R p4 , R p5 Each is independently selected from C1-C10 alkyl, C1-C10 alkoxy, C3-C8 cycloalkyl or C2-C8 heterocycloalkyl; (4) Zwitterionic derivatives or zwitterionic polymers: in, pd1 is selected from an integer between 5 and 250; R p6 Selected from: pc1, pe1 are integers between 0 and 10, A - Selected from R p3 , R p4 , R p5 Each is independently selected from C1-C10 alkyl, C1-C10 alkoxy, C3-C8 cycloalkyl or C2-C8 heterocycloalkyl; L p is absent or a connecting unit selected from alkanes, heterocycloalkanes, aromatic hydrocarbons, heterocycloalkanes or a combination thereof; (5) Polysaccharide polymers: Among them, pf1, pf2, pg1, pg2, ph1, ph2, pi1, pi2 and m are selected from integers between 1-250, preferably integers between 1-50, for example, integers between 1-5, integers between 4-24, or integers between 5-25.

8. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claims 1-4, characterized in that: The bioactive molecule fragment D is selected from: small molecule drugs, peptides, proteins, nucleic acids, and carbohydrates, and specifically can be anesthetics, ophthalmic drugs, central nervous system drugs, pain therapeutic agents, antiarthritis drugs, anticonvulsants, antihistamines, antiulcer drugs, behavior correction drugs, respiratory drugs, antispasmodics, muscle relaxants, anti-inflammatory drugs, appetite suppressants, growth promoters, hemostatics, immunostimulants, immunomodulators, muscle relaxants, obesity therapeutic agents, osteoporosis drugs, tranquilizers, tranquilizers, migraine drugs, muscle contraction drugs, anti-infective drugs, anti-rheumatic drugs, antimalarial drugs, Antiemetics, bronchodilators, antithrombotics, antihypertensives, cardiovascular drugs, antiarrhythmics, antioxidants, antiasthmatics, diuretics, lipid regulators, antiandrogens, antiparasitics, anticoagulants, antitumor drugs, hypoglycemics, nutritional agents, growth supplements, anti-enteritis agents, vaccines, antibodies, radionuclides, diagnostic agents and contrast agents; preferably, selected from small molecule drugs or peptide drugs; more preferably, selected from antitumor drugs, such as cytotoxic drugs, small molecule targeted drugs, hormones, biological response modifiers (such as immunomodulators), tumor adjuvant drugs and radionuclide drugs.

9. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claims 1-8, characterized in that: The phospholipid drug conjugate has a structure shown in formula (II): Among them, X 0 , X 1 , L 1 , P 1 , X 2 , L 2 , Y 2 , Q 2a , Q 2b , Y 2 , D as described in claims 1-7; Where T 0 It is present or absent. When present, the connection point with the P terminal is selected from Where R 01 is hydrogen, C1-C10 alkyl, C1-C10 alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl; L is L 3 -L s , L 3 is an environmentally responsive connection unit, L s It exists or does not exist. When it exists, it is a self-releasing connecting part; when T 0 When it does not exist, L 3 It is directly connected to the P end, and the functional group at the connection point is a nitrogen atom.

10. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claim 9, characterized in that: X 0 , X 1 , X 2 is O; L 1 , L 2 Select from absent or present, and when present, from: For the P terminal connection, P 1 or Y 2 End connection; l1a is selected from an integer between 0-10, preferably an integer between 1-5; Y 2 Selected from, Preferably, Y 2 Selected from More preferably, Y 2 Selected from For L 2 End connection; P 1 Selected from the group consisting of pa1 repeating units and R p1 A polyethylene glycol derivative residue, wherein pa1 is selected from an integer between 10 and 60, preferably an integer between 15 and 50, such as 21, 22, 44 or 45; R p1 is a C1-C6 alkyl group, preferably a C1-C3 alkyl group, such as methyl, ethyl, n-propyl or isopropyl; or P 1 Selected from the group consisting of pb1 repeating units and R p2 Polycreatine derivative residues: wherein pb1 is selected from an integer between 10 and 60, preferably an integer between 15 and 50; R p2 is selected from hydrogen, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl or an amine protecting group; Q 2a T 2a -R 1 , Q 2b T 2b -R 2 , T 2a 、T 2b As defined in claim 1, R 1 , R 2 Selected from: Wherein, r1a and r1b are selected from integers of 1-30.

11. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claim 1-3 or 8, characterized in that: The biologically active molecule fragment D is selected from: (1) Cytotoxic drugs: Camptothecins, a topoisomerase inhibitor represented by formula (III): Among them, R x1 is selected from C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 cycloalkyl, C1-C4 deuterated alkyl, C2-C4 alkynyl or C2-C4 alkenyl; R x2 , R x3 , R x4 and R x5 Each is independently selected from a hydrogen atom, a deuterium atom, a halogen, a cyano group, a hydroxyl group, a thiol group, a sulfone group, a sulfoxide group, a substituted or unsubstituted amino group, a nitro group, an alkynyl group, an alkenyl group, an alkyl group, a haloalkyl group, a cycloalkyl group, an alkoxy group, an alkylthiol group, a heterocycloalkyl group, a deuterated alkyl group, an aryl group or a heteroaryl group; Or, R x1 and R x2 , R x2 and R x3 , R x3 and R x4 , R x4 and R x5 Each together with the carbon atom to which they are commonly attached forms a 5-7 membered cycloalkyl or heterocycloalkyl group; X is selected from O or S; Z is selected from OH or F; n1 is selected from 0 or 1; Preferably, it is selected from the following structures: Tubulin inhibitors are preferably selected from: Nucleic acid transcription, preferably, is selected from: Drugs acting on DNA structures are preferably selected from: Among them, R t1 is selected from hydrogen, alkyl, alkoxy, heteroalkyl, aryl or heteroaryl; (2) Small molecule targeted drugs: EGFR inhibitors, such as gefitinib, erlotinib, icotinib, afatinib, or osimertinib; HER2 inhibitors, such as lapatinib, pyrotinib, neratinib, or tucatinib; ALK inhibitors, such as crizotinib, alectinib, ceritinib, lorlatinib, or brigatinib; MEK inhibitors, such as trametinib, cabozantinib, or bimetinib; MET inhibitors, such as savotinib, savolitinib, or capmatinib; BRAF inhibitors, such as sorafenib, dabrafenib, vemurafenib, vemurafenib, or contrafenib; CDK4 / 6 inhibitors, such as palbociclib, ribociclib, abemaciclib, or PARP inhibitors, such as olaparib, niraparib, rucaparib, or talazoparib; BTK inhibitors, such as acalabrutinib, ibrutinib, zanubrutinib, tirabrutinib, or zanubrutinib; Protease inhibitors, such as bortezomib, carfilzomib, or ixazomib; mTOR inhibitors, such as temsirolimus or everolimus; JAK inhibitors, such as ruxolitinib, tofacitinib, upadacitinib, baricitinib, digoxinib, piricitinib, filgotinib, abrocitinib, paclitinib, or deuterocelexinib; PI3K inhibitors, such as idelalisib, panisib, duvelisib, or upalisib; VEGFR inhibitors, such as sorafenib, axitinib, apatinib, sunitinib, regorafenib, vandetanib, pazopanib, lenvatinib, cabozantinib, ponatinib, aflibercept, or fruquintinib; PDGFR, c-Kit inhibitors, such as imatinib, nilotinib, or afatinib; BCL inhibitors, such as venetoclax; HDAC inhibitors, such as vorinostat, romidepsin, belinostat, panobinostat, or chidamide; FLT3 inhibitors, such as midostaurin, gilteritinib, or quizartinib; RET inhibitors, such as sepretib or pralsetinib; KRAS inhibitors, such as sotolacizumab or adagracilib; IDH inhibitors, such as emdesipine, ilotinib, or olutacinib; BCR-ABL inhibitors, such as imatinib, dasatinib, nilotinib, ponatinib, or orebactinib; (3) hormonal molecules, such as fulvestrant, abiraterone, prednisone, dexamethasone, bicalutamide, enzalutamide, tamoxifen, toremifene, letrozole, exemestane, goserelin, or leuprolide; (4) Immunotherapy molecules, such as small molecule PD-L1 inhibitors, TLR agonists, stimulator of interferon genes (STING) agonists, or indoleamine-2,3-dioxygenase (IDO) inhibitors; (5) Other small molecules and peptides, such as lenalidomide, thalidomide, pomalidomide, ibedomide, octreotide, lanreotide or pasireotide.

12. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claims 1-11, characterized in that: LD is selected from any one of the following conditions: (1) When the drug molecule D is of the following structure, the connection site with L is express: L is selected from the following structures, and its connection site with drug D is express: (2) When the drug molecule D is of the following structure, the connection site with L is express: L is selected from the following structures, and its connection site with drug D is express:

13. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claims 9 to 12, characterized in that: The phospholipid drug conjugate has a structure shown in formula (IV): in, (1) Selected from the following structures: Wherein, x and q are selected from integers between 10 and 60; pf1, pf2, ph1, ph2, pi are selected from integers between 1 and 100; (2) Selected from the following structures: Wherein, y, z, y1, y2, z1, z2, z3 are selected from integers of 1-20; (3) Selected from the following structures:

14. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claim 1, characterized in that: It meets one or more of the following conditions: (1)X 0 is O; (2)X 1 is O; (3)X 2 is O; (4)M 0 -T 0 -(LD) n , preferably -LD, n is preferably 1; (5)T 0 is absent or present. When present, T 0 Preferably selected from -NHCH2- and wherein the C-terminus is preferably connected to L; (6)M 2 for not existing; (7)L 1 for Preferably With X 1 connect, With P 1 connection, l1a is selected from an integer between 0 and 10, preferably an integer between 0 and 5, such as 1 or 2; (8)L 2 for With X 1 connect, With P 1 Connection; l1a is selected from an integer between 0-10, preferably an integer between 0-5, such as 0 or 2; (9)K 1 P 1 , P 1 The pharmacokinetic regulating part unit is selected from the group consisting of polyethylene glycol derivative residues, polycreatine derivative residues, quaternary ammonium salt derivative residues, zwitterion derivatives or zwitterion polymers, and polysaccharide polymers, preferably selected from the group consisting of polyethylene glycol derivative residues, polycreatine derivative residues, quaternary ammonium salt derivative residues and polysaccharide polymers, more preferably polyethylene glycol derivative residues; the polyethylene glycol derivative residue is preferably pa1 is selected from an integer between 5 and 250, preferably selected from an integer between 5 and 150 (e.g., 22, 45 or 113), more preferably selected from an integer between 10 and 60 (e.g., 45); pr1 is selected from an integer between 0 and 8, preferably selected from an integer between 0 and 2, such as 0; R p1 is hydrogen, C1-C10 alkyl, C1-C10 heteroalkyl, C3-C10 cycloalkyl, C3-C10 alkenyl, C3-C10 alkynyl or a hydroxyl protecting group, preferably hydrogen, C1-C10 alkyl or a hydroxyl protecting group, more preferably C1-C10 alkyl; the polycreatine derivative residue is preferably pb1 is selected from an integer between 5 and 250, preferably selected from an integer between 5 and 150, and more preferably selected from an integer between 10 and 50, for example, 28; R p2 is selected from hydrogen, C1-C10 alkyl, C1-C10 alkoxy, C3-C10 alkenyl, C3-C10 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C10 aryl, C5-C10 heteroaryl and amine protecting group, preferably amine protecting group, such as -C(O)CH3; the quaternary ammonium salt derivative residue is preferably R p3 , R p4 , R p5 Each is independently selected from C1-C10 alkyl, C1-C10 alkoxy, C3-C8 cycloalkyl or C2-C8 heterocycloalkyl, preferably C1-C10 alkyl, such as methyl; the polysaccharide polymer is preferably selected from: wherein pf1, pf2 and m are each independently selected from an integer between 1 and 250, preferably an integer between 1 and 50, such as an integer between 1 and 5, an integer between 4 and 24, or an integer between 5 and 25; (10)Y 2 is a branching center, selected from Preferably selected from More preferred For L 2 End connection; (11)T 2a 、T 2b is absent or present. When present, T 2a 、T 2b are each independently selected from -C(O)-; (12)R 1 and R 2 In the above, C1~C 50 The alkyl groups are each independently C8 to C 50 Alkyl, more preferably C 10 ~C 30 Alkyl, more preferably C 10 ~C 20 Alkyl; preferably, R 1 and R 2 Each independently selected from Preferably selected from wherein r1a and r1b are selected from integers of 1 to 30, Optionally, one or more selected from hydroxy, halogen, C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 The substituents of cycloalkyl and 3-12 membered heterocycloalkyl are substituted, the C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 Cycloalkyl and 3-12 membered heterocycloalkyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-6 Substitution of cycloalkyl groups; Or, R 1 and R 2 Each independently preferably selected from More preferably selected from Wherein, r1a and r1b are selected from integers of 1-30. Optionally, one or more selected from hydroxy, halogen, C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 The substituents of cycloalkyl and 3-12 membered heterocycloalkyl are substituted, the C 1-10 Alkyl, C 1-10 Alkoxy, C 3-12 Cycloalkyl and 3-12 membered heterocycloalkyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-6 Substitution of cycloalkyl groups; (13) The bioactive molecule fragment D is preferably selected from cytotoxic drugs, small molecule targeted drugs, hormone molecules, and other small molecules and polypeptides; The cytotoxic drugs are preferably selected from the topoisomerase inhibitors camptothecins, microtubule inhibitors and drugs acting on DNA structures as shown in formula (III), and more preferably selected from the topoisomerase inhibitors camptothecins and microtubule inhibitors as shown in formula (III); The camptothecin class of topoisomerase inhibitors represented by formula (III) is preferably selected from: The microtubule inhibitors are preferably The drugs acting on DNA structure are preferably selected from wherein Rt1 is selected from hydrogen, alkyl, alkoxy, heteroalkyl, aryl and heteroaryl; The small molecule targeted drugs are preferably selected from ALK inhibitors, CDK4 / 6 inhibitors and PARP inhibitors; the ALK inhibitor is preferably crizotinib; the CDK4 / 6 inhibitor is preferably selected from palbociclib and The PARP inhibitor is preferably niraparib; The hormone molecule is preferably fulvestrant; The other small molecules and polypeptides are preferably octreotide.

15. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claim 1, characterized in that: It meets one or more of the following conditions: (1)R 1 and R 2 Each independently selected from Preferably selected from Or, R 1 and R 2 Each independently preferably selected from More preferably (2)Q 2a and Q 2b Each independently selected from Preferably selected from Or, Q 2a and Q 2b Each independently preferably selected from More preferably selected from (3) D is selected from the following structures: (4) L is selected from the following structures (the connection site with D is express): When L is When T 0 Preferably -NHCH2-, wherein the C-terminus is connected to L; when L is When T 0 Preferably The C-terminus is connected to L; when L is other fragments, T 0 Preferably, it is absent.

16. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claim 1, characterized in that: It meets one or more of the following conditions: (1) Selected from the following structures: (2) Selected from the following structures: (3) Selected from the following structures:

17. The phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate according to claims 1-16, characterized in that: The phospholipid drug conjugate is selected from the following structures:

18. Use of the phospholipid drug conjugate or its stereoisomer, prodrug, pharmaceutically acceptable salt or pharmaceutically acceptable solvate as described in claims 1-17, or the pharmaceutical composition in a drug for preventing, alleviating and / or treating abnormal cell proliferation, infection (e.g., viral infection, such as HIV, HBV, etc.), inflammatory disease (e.g., rheumatoid arthritis, etc.), autoimmune disease (e.g., psoriasis, lupus erythematosus, diabetes, etc.), cardiovascular disease (e.g., stroke, myocardial infarction, atherosclerosis, etc.), eye disease or neurodegenerative disease (e.g., Alzheimer's disease, Parkinson's disease, etc.), wherein the abnormal cell proliferation disease can be cancer; the cancer preferably includes breast cancer, ovarian cancer, brain cancer, prostate cancer, melanoma, nasopharyngeal cancer, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer (e.g., colon cancer, pancreatic cancer, colorectal cancer, etc.), cerebral infarction, encephalopathy, leukemia, leukemia, necrosis ... Intestinal cancer, rectal cancer, etc.), lung cancer (such as small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma, poorly differentiated carcinoma, etc.), kidney cancer, skin cancer, neuroblastoma, sarcoma, liposarcoma, osteochondroma, bone cancer, osteosarcoma, seminoma, testicular tumor, uterine tumor (such as cervical cancer, endometrial cancer, etc.), head and neck tumor (such as laryngeal cancer, pharyngeal cancer, tongue cancer, etc.), multiple myeloma, malignant lymphoma (such as reticular cell sarcoma, palatine lymphosarcoma, Hodgkin lymphoma, mantle cell lymphoma, etc.), polycythemia vera, leukemia (such as acute myeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, etc.), thyroid tumor, ureter tumor, bladder tumor, gallbladder cancer, bile duct cancer, choriocarcinoma or pediatric tumor (such as neuroblastoma, embryonic testicular cancer, retinoblastoma, etc.).

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