Novel GalNAc targeted delivery fragment and preparation and application thereof

Abstract

The invention discloses a GalNAc delivery molecule shown in the following formula (I), or a stereoisomer, an enantiomer or a pharmaceutically acceptable salt thereof, wherein the definition of each group is described in the specification. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular, provides a novel GalNAc targeted delivery fragment and its preparation and application. Background Art

[0002] Hepatitis B virus (HBV) belongs to the Hepaciviridae family of small, enveloped, primarily hepatotropic viruses with a partially double-stranded DNA genome of approximately 3.2 kb in size. HBV infection remains a major public health problem.

[0003] Current treatments are limited to two classes of drugs, which are nucleoside (nucleotide) analogs (lamivudine, adefovir, tenofovir, telbivudine, and entecavir) and interferon alpha (INF-α, both non-PEGylated and PEGylated). Although both therapies can reduce HBV DNA and normalize liver enzymes, neither therapy can provide a high level of clinical cure, which is defined by the loss of HBV surface antigen (HBsAg) with or without seroconversion. Interferon-based therapies are poorly tolerated and are only effective against certain viral genotypes. Although nucleoside (nucleotide) analogs often require prolonged or potentially lifelong treatment, and some have resistance, low efficacy, and tolerability issues. Therefore, there is still a huge medical need to discover and develop effective and safe anti-HBV drugs with novel mechanisms of action to improve disease cure rates. (Marcellin, Patrick, et al. The Lancet 381.9865 (2013): 468-475; Tang, Lydia SY, et al. Jama 319.17 (2018): 1802-1813.)

[0004] GalNAc-mediated liver-targeted delivery of small interfering RNA drugs (GalNAc-siRNA) can inhibit the expression of target genes at the basic level of mRNA translation into protein, and has recently been used to treat a variety of liver-related diseases. GalNAc-siRNA molecules have the characteristics of strong efficacy, high liver targeting specificity, and good safety, so they have unique advantages in the field of anti-hepatitis B, but current experimental drugs cannot effectively reduce HBsAg content and cannot achieve a high level of clinical cure rate. Therefore, it is necessary to optimize this type of molecule, focusing on the structural modification of the targeted delivery fragment containing GalNAc to improve delivery efficiency and drug properties. Summary of the invention

[0005] The object of the present invention is to provide a GalNAc-siRNA molecule and a nucleic acid delivery pharmaceutical composition prepared using the same.

[0006] In the first aspect of the present invention, there is provided a GalNAc delivery molecule as shown in the following formula (I), or a stereoisomer, enantiomer, or a pharmaceutically acceptable salt thereof:

[0007]

[0008] in,

[0009] Ax is a substituted or unsubstituted 4-12-membered saturated or partially unsaturated nitrogen-containing heterocyclic group, or a substituted or unsubstituted 4-12-membered nitrogen-containing heteroaryl group;

[0010] R1 is selected from the group consisting of H, DMTr, phosphate, phosphorothioate, phosphorothioate-siRNA conjugate, or phosphate-siRNA conjugate;

[0011] Linker is a divalent linking group;

[0012] Q is selected from the group consisting of -NH-C(O)-, -C(O)-;

[0013] r is 0, 1, 2, 3, 4, 5 or 6;

[0014] R2 is a GalNAc target head portion and has a structure selected from the group consisting of:

[0015] Wherein, L1 and L2 are each independently a trivalent linking group; L3, L4, L5, and L6 are divalent linking groups, and R3, R4, and R5 are each independently a substituted or unsubstituted sugar group, preferably a group formed by a substituted or unsubstituted N-acetylgalactosamine molecule;

[0016] Selected from the group consisting of: substituted or unsubstituted C 4-15 a saturated or partially unsaturated carbon ring group, a substituted or unsubstituted 4-12 membered saturated or partially unsaturated nitrogen-containing heterocyclic group, a substituted or unsubstituted C 6-18 Aryl, substituted or unsubstituted 5-18 membered heteroaryl;

[0017] The substitution refers to the substitution of one or more hydrogen atoms on the group by a substituent selected from the following group: 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, 3- to 12-membered heterocyclic group, C 3-8 Aryl, 5- to 7-membered heteroaryl, halogen, hydroxyl, carboxyl (-COOH), C 1-8 Aldehyde, C 2-10 Acyl, C 2-10 Ester group, amino group, C 1-8 Alkoxy, C 1-10or two substituents located on adjacent ring atoms may together with the connected ring atoms form a group selected from the following group: a 5-7 membered carbon ring or heterocyclic ring, a benzene ring, or a 5-7 membered heteroaromatic ring.

[0018] In another preferred embodiment, Having a structure selected from the group consisting of:

[0019]

[0020] Wherein, R' is selected from the following group: H or C 1-8 alkyl;

[0021] p is 1, 2, 3, 4, 5 or 6.

[0022] In another preferred embodiment, the siRNA is selected from the following group: siRNA that inhibits the expression of hepatitis B virus (HBV) gene, siRNA that inhibits the expression of apolipoprotein C3 (APOC3) gene, siRNA that inhibits the expression of PCSK9 gene, siRNA that inhibits the expression of CCR4 gene, and siRNA that inhibits the expression of thyroxine transporter (TTR) gene.

[0023] In some embodiments, the Linker has a structure selected from the following group:

[0024] C4-C20 alkylene,

[0025] wherein n, n3 and m are each independently selected from the following group: 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12;

[0026] n1 and n2 are each independently selected from the following group: 0 or 1;

[0027] Z is selected from the group consisting of a single bond, substituted or unsubstituted C 1-8 Alkylene, substituted or unsubstituted 7-12 membered fused bicyclic ring, substituted or unsubstituted 7-15 membered spirocyclic ring, substituted or unsubstituted 5-15 membered bridged ring, -NH-substituted or unsubstituted 7-12 membered fused bicyclic ring, -NH-substituted or unsubstituted 7-15 membered spirocyclic ring, -NH-substituted or unsubstituted 5-15 membered bridged ring, -NH-substituted or unsubstituted 7-12 membered fused bicyclic ring-NH-, -NH-substituted or unsubstituted 7-15 membered spirocyclic ring-NH-, -NH-substituted or unsubstituted 5-15 membered bridged ring-NH-; wherein the ring atoms of the fused bicyclic ring, spirocyclic ring and bridged ring may be optionally carbon atoms or heteroatoms;

[0028] The substitution refers to the substitution of one or more hydrogen atoms on the group by a substituent selected from the following group: 1-8 Alkyl, C 2-8 Alkenyl, C2-8 Alkynyl, C 3-8 Cycloalkyl, 3- to 12-membered heterocyclic group, C 6-10 Aryl, 5- to 10-membered heteroaryl, halogen, hydroxyl, carboxyl (-COOH), C 1-8 Aldehyde, C 2-10 Acyl, C 2-10 Ester group, amino group, C 1-8 Alkoxy, C 1-10 Sulfonyl; or two substituents located on adjacent ring atoms may together with the connected ring atoms form a group selected from the following group: a 5-7 membered carbon ring or heterocyclic ring, a benzene ring, or a 5-7 membered heteroaromatic ring;

[0029] Alternatively, Z has a structure selected from the group consisting of:

[0030]

[0031] wherein s1, s2, s, s4, s5 and s6 are each independently selected from the following group: 0, 1 or 2;

[0032] s7 is selected from the group consisting of 1, 2, 3, 4, 5 or 6;

[0033] Each of M1, M2, M3 and M4 is independently selected from the group consisting of CHR, C(R)R, C(O), O, S, NR;

[0034] M5 and M6 are each independently selected from the following group: (CHR) s6 、C(R)R、C(O)、(CHR) s6 -C(O), O, S, NR;

[0035] R is selected from the following group: H, halogen, methyl; or two R located on adjacent reducing atoms and the carbon atoms to which they are connected together constitute a substituted or unsubstituted C 3-8 Saturated or partially unsaturated carbocyclic ring, 4-8 membered saturated or partially unsaturated heterocyclic ring, C 6-10 Aromatic ring or 5-12 membered heteroaromatic ring.

[0036] In some embodiments, the Z has a structure selected from the following group:

[0037]

[0038] In some embodiments, L1, L2 and L3 are each independently a structure shown in the following formula:

[0039]

[0040] The L3, L4, L5 and L6 are each independently a structure shown in the following formula:

[0041]

[0042] Among them, R 11 Selected from the following group: H or C 1-4 alkyl;

[0043] Each u, v and w is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11; Glu is an unmodified or modified 5-6-membered sugar group;

[0044] X1 is selected from the following group: substituted or unsubstituted C 4-10 a saturated or partially unsaturated carbocyclic group, a substituted or unsubstituted 4-10 membered saturated or partially unsaturated heterocyclic group (preferably a nitrogen-containing heterocyclic group), a substituted or unsubstituted C 6-10 Phenyl, or substituted or unsubstituted 5-10 membered heteroaryl;

[0045] X2 is selected from the following groups: -NH-(C1-C6 alkyl)-NH-, -NH-(C3-C8 cycloalkyl)-NH-, -NH-(4-10 membered heterocyclyl)-NH-, substituted or unsubstituted C 4-10 a saturated or partially unsaturated carbocyclic group, a substituted or unsubstituted 4-14 membered saturated or partially unsaturated heterocyclic group (preferably a nitrogen-containing heterocyclic group), a substituted or unsubstituted C 6-10 Phenyl, or substituted or unsubstituted 5-10 membered heteroaryl;

[0046] The carbocyclic group and heterocyclic group may be a single ring, a condensed ring, a bridged ring or a spiro ring.

[0047] In some embodiments, the X1 is a substituted or unsubstituted structure selected from the following group:

[0048]

[0049] wherein s1, s2, s, s4, s5 and s6 are each independently selected from the following group: 0, 1 or 2;

[0050] s7 is selected from the group consisting of 1, 2, 3, 4, 5 or 6;

[0051] Each of M1, M2, M3 and M4 is independently selected from the group consisting of CHR, C(R)R, C(O), O, S, NR;

[0052] M5 and M6 are each independently selected from the following group: (CHR) s6 、C(R)R、C(O)、(CHR) s6 -C(O), O, S, NR;

[0053] R is selected from the following group: H, halogen, methyl; or two R located on adjacent reducing atoms and the carbon atom to which they are connected together form a substituted or unsubstituted 4-8 membered carbocyclic or heterocyclic ring (including saturated, unsaturated or aromatic ring).

[0054] In another preferred embodiment, the X2 is a substituted or unsubstituted structure selected from the following group:

[0055]

[0056] In some embodiments, the sugar group has a structure as shown in Formula III below:

[0057]

[0058] Among them, the R 15 Selected from the group consisting of -NH(C2-C6 acyl), -NH(halogenated C2-C6 acyl), -NH(C2-C6 sulfonyl), -NH(halogenated C2-C6 sulfonyl);

[0059] R 12 , R 13 and R 14 Each is independently selected from the following group: H or C1-C6 acyl.

[0060] In some embodiments, the R 15 Selected from the group consisting of: -NH(C2-C4 acyl); R 12 , R 13 and R 14 Each is independently selected from the following group: H or C1-C4 acyl.

[0061] In some embodiments, the R 15 is -NHAc; R 12 , R 13 and R 14 Each is independently selected from the group consisting of H or Ac.

[0062] In some embodiments, the is a substituted or unsubstituted structure selected from the following group: a substituted or unsubstituted 9-18 membered nitrogen heterocycle, a substituted or unsubstituted C 4-12 Monocycloalkyl, substituted or unsubstituted C6-15 bridged cycloalkyl.

[0063] In another preferred embodiment, the is a substituted or unsubstituted structure selected from the following group:

[0064] In some embodiments, the GalNAc delivery molecule has a structure selected from the group consisting of:

[0065]

[0066]

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080] In some embodiments, the GalNAc delivery molecule has a structure selected from the group consisting of:

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089] In the second aspect of the present invention, a pharmaceutical composition is provided, characterized in that the pharmaceutical composition comprises: one or more of the compound of formula I as described in the first aspect of the present invention, its pharmaceutically acceptable salt, racemate, R-isomer, S-isomer or mixture thereof, and one or more pharmaceutically acceptable carriers, excipients, adjuvants, auxiliary materials and / or diluents.

[0090] In some embodiments, the composition is used to prepare a pharmaceutical composition for treating or preventing tumors or infections caused by viruses.

[0091] In some embodiments, the invention is used to prepare a medicament for treating and / or preventing a disease or condition selected from the following groups: a disease or condition caused by hepatitis B virus, a disease or condition caused by abnormal expression of ANGPTL3 gene, a disease or condition caused by abnormal expression of TTR gene, a disease or condition caused by abnormal expression of hepatocyte gene, or other liver disease or condition.

[0092] In the third aspect of the present invention, there is provided a GalNAc delivery molecule linker as shown in formula (Ia), or a stereoisomer, enantiomer, or a pharmaceutically acceptable salt thereof:

[0093]

[0094] The part shown in formula (Ia) has a structure selected from the group consisting of

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104] in, It is the site connected to R1.

[0105] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0106] Based on long-term and in-depth research, the inventors have prepared a novel type of GalNAc targeted drug delivery fragment with a novel structure. The drug or drug composition composed of the GalNAc targeted delivery fragment has good targeted delivery effect, good efficacy and higher safety. Based on the above findings, the inventors have completed the present invention.

[0107] the term

[0108] In the present invention, the halogen is F, Cl, Br or I.

[0109] In the present invention, unless otherwise specified, the terms used have the general meanings well known to those skilled in the art.

[0110] In the present invention, the term "C1-C6 alkyl" refers to a straight or branched alkyl group having 1 to 6 carbon atoms, including but not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexyl, etc.; preferably ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl.

[0111] In the present invention, the term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, isopropoxy, butoxy and the like.

[0112] In the present invention, the term "C3-C7 cycloalkyl" refers to a cyclic alkyl group having 3 to 7 carbon atoms in the ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. The terms "C5-C6 cycloalkyl" and "C3-C6 cycloalkyl" have similar meanings.

[0113] In the present invention, the term "aromatic ring" or "aryl group" has the same meaning, preferably "aryl group" is "C6-C 12 Aryl" or "C6-C 10 The term "C6-C 12 "Aryl" refers to an aromatic ring group having 6 to 12 carbon atoms without heteroatoms in the ring, such as phenyl, naphthyl, etc. The term "C6-C 10 "Aryl" has a similar meaning.

[0114] In the present invention, the term "aromatic heterocycle" or "heteroaryl" has the same meaning and refers to a heteroaromatic group containing one to multiple heteroatoms. The heteroatoms referred to here include oxygen, sulfur and nitrogen. For example, furanyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring can be fused to an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. The heteroaryl group can be optionally substituted or unsubstituted.

[0115] In the present invention, the term "3-9 membered carbocyclic group" refers to a saturated or unsaturated (non-aromatic ring, including monocyclic, cyclic, spirocyclic, bridged ring, etc.) 3-9 membered cyclic group, whose ring skeleton structure only includes carbon atoms, such as cyclopentyl, cyclohexyl, etc.

[0116] In the present invention, the term "3-9 membered heterocyclic group" refers to a 3-9 membered cyclic group containing 1 to 3 heteroatoms selected from oxygen, sulfur and nitrogen, which is saturated or unsaturated (non-aromatic ring, including monocyclic, cyclic, spirocyclic, bridged, etc.), such as dioxolanyl, etc. The term "3-7 membered heterocyclic group" has a similar meaning.

[0117] In the present invention, the term "substituted" refers to one or more hydrogen atoms on a specific group being replaced by a specific substituent. The specific substituent is a substituent described above or a substituent appearing in the embodiments. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable position of the group, and the substituent may be the same or different at each position. A cyclic substituent, such as a heterocycloalkyl, may be connected to another ring, such as a cycloalkyl, to form a spirobicyclic system, for example, the two rings have a common carbon atom. It will be understood by those skilled in the art that the combinations of substituents contemplated by the present invention are those that are stable or chemically feasible. The substituents are, for example (but not limited to): C 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, 3- to 12-membered heterocyclic group, aryl, heteroaryl, halogen, hydroxyl, carboxyl (-COOH), C 1-8 Aldehyde, C 2-10 Acyl, C 2-10 Ester group, amino group, alkoxy group, C 1-10 Sulfonyl, etc.

[0118] Unless otherwise stated, where substituents are designated by their conventional chemical formula (written from left to right), they equivalently include chemically identical substituents resulting from the structure written from right to left, e.g., -NH-C(O)- is intended to include -C(O)-NH-, and -COO- is intended to include -OCO-.

[0119] Pharmaceutical composition containing active ingredients

[0120] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention and a pharmaceutically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 0.01-99.99% by weight of the compound of the present invention / dose, and more preferably, contains 0.1-99.9% by weight of the compound of the present invention / dose. Preferably, the "one dose" is a capsule or tablet.

[0121] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0122] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral administration, parenteral administration (intravenous administration, intramuscular administration or subcutaneous administration).

[0123] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0124] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0125] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0126] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0127] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0128] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0129] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds (eg, anti-HBV agents).

[0130] When administered in combination, the pharmaceutical composition also includes one or more (2, 3, 4, or more) other pharmaceutically acceptable compounds. One or more (2, 3, 4, or more) of the other pharmaceutically acceptable compounds can be used simultaneously, separately or sequentially with the compound of the present invention to prevent and / or treat related diseases or conditions.

[0131] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage when applied is a pharmaceutically effective dosage. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health status, which are all within the skill of a skilled physician.

[0132] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples that do not specify specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0133] In the following examples, the structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of (ppm) is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD) as the measuring solvent, and tetramethylsilane (TMS) as the internal standard.

[0134] SHIMADZU LC system (chromatographic column: CSH TM Prep-C18, 19*150mm, liquid handler LH-40, pump LC-20AP, detector SPD-20A, system controller CBM-20A, solvent system: acetonitrile and 0.05% trifluoroacetic acid aqueous solution).

[0135] LC / MS spectra of the compounds were obtained using LC / MS (Agilent Technologies 1200 Series). LC / MS conditions were as follows (run time was 10 minutes):

[0136] Acidic conditions: A: 0.05% trifluoroacetic acid in water; B: 0.05% trifluoroacetic acid in acetonitrile;

[0137] Alkaline conditions: A: 0.05% NH3·H2O aqueous solution; B: acetonitrile

[0138] Neutral conditions: A: 10 mM NH4OAC in water; B: acetonitrile

[0139] Unless otherwise specified, in the following examples, intermediates and final compounds were purified by silica gel column chromatography or by Xselect ○ R CSH TM Prep-C18(5μm, OBD TM19*150mm) column or XBridgeTM PrepPhenyl (5μm, OBD TM 30*100 mm) was purified by preparative HPLC on a reverse phase column.

[0140] Silica gel column chromatography generally uses Yantai Huanghai Silica Gel 200-300 mesh silica gel as the carrier.

[0141] The CombiFlash rapid preparation instrument used was Combiflash Rf200 (TELEDYNE ISCO).

[0142] Thin layer chromatography (TLC) silica gel plates use Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The specifications of silica gel plates used in thin layer chromatography detection products are 0.15mm~0.2mm, and the specifications used in thin layer chromatography separation and purification products are 0.4mm~0.5mm.

[0143] The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, Darui Chemicals and other companies.

[0144] Abbreviations:

[0145] TMSOTf: trimethylsilyl trifluoromethanesulfonate; NaIO4: sodium periodate; RuCl3: ruthenium trichloride; BnBr: benzyl bromide; K2CO3: potassium carbonate; DIEPA (DIEA): N,N-diisopropylethylamine; TFA: trifluoroacetic acid; DCM: dichloromethane; TEA: triethylamine; HATU: benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate; CbzCl: benzyl chloroformate; NaOH: sodium hydroxide; DMSO: dimethyl sulfoxide; HOBt: 1-hydroxybenzotriazole; EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; DMF: N,N-dimethylformamide; tert-butyl acrylate: tert-butyl acrylate; Formic acid: formic acid; DMTr: 4,4'-dimethoxytrityl; Ac: acetyl; Boc: tert-butyloxycarbonyl; Bn: benzyl; Cbz: benzyloxycarbonyl

[0146] Preparation Example Preparation of Compound A1

[0147]

[0148] Step 1: Preparation of compound A1-1

[0149] 4A molecular sieve (30 g) and TMSOTf (18.8 g, 84.8 mmol) were slowly added to a dichloromethane (300 ml) solution of compound Int.1 (30 g, 77.1 mmol); after the addition, the reaction mixture was stirred at 55°C for 3 hours. Then the reaction mixture was neutralized with triethylamine, poured into 500 ml of water, and diluted and extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow oil compound A1-1 (22.8 g). MS: 330.2 (M+H) + .

[0150] Step 2: Preparation of Compound A1-2

[0151] Under nitrogen protection, 4A molecular sieve (10 g) and compound Int.02 (3.7 g, 15.2 mmol) were slowly added to a dichloroethane (100 ml) solution of compound A1-1 (11 g, 30.4 mmol). After the addition, the reaction mixture was stirred at 25°C for 30 minutes; then TMSOTf (3.7 g, 15.2 mmol) was slowly added to the above reaction mixture under stirring at 0 degrees, and the reaction mixture was stirred at 25°C for 16 hours. The reaction mixture was filtered, the filter cake was washed with dichloromethane, the filtrate was neutralized with sodium bicarbonate solution and extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a yellow oil compound A1-2 (7 g, 48.8%). MS: 430.2 (M+H) + .

[0152] Step 3: Preparation of Compound A1-3

[0153] Under nitrogen protection, a mixed solution of NaIO4 (13.0 g, 60.6 mmol) and compound A1-3 (6.5 g, 15.2 mmol) in dichloromethane (30 ml), acetonitrile (30 ml) and water (45 ml) was stirred at 0°C for 30 minutes; then RuCl3 3H2O (95 mg, 0.45 mmol) was slowly added to the above reaction mixture under stirring at 0 degrees, and the reaction mixture was stirred at 25°C for 16 hours. The reaction mixture was diluted with 100 ml of water and sodium bicarbonate was added to adjust the pH value to 7.5, and extracted with dichloromethane. The aqueous phase was adjusted to pH 3 with citric acid and extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a white foam solid compound A1-3 (3.2 g, 45%). MS: 448.2 (M+H) + . 1H NMR (400MHz, DMSO-d6) δ11.98(s,1H),7.81(d,J=9.2Hz,1H),5.21(d,J=3.2Hz,1H),4.96(dd,J=11.2,3.4Hz,1H),4.49(d,J=8.4Hz,1H),4.02(s, 3H),3.87(d,J=10.0Hz,1H),3.78–3.37(m,2H),2.20(t,J=6.8Hz,2H),2. 10(s,3H),2.00(s,3H),1.89(s,3H),1.77(s,3H),1.49(d,J=2.8Hz,4H).

[0154] Step 4: Preparation of Compound A1

[0155] Under nitrogen protection at 0 degrees, compound Int.3 (10.3 g, 36.8 mmol) was slowly added to a solution of compound A1-3 (11.0 g, 24.6 mmol) and DIEA (6.4 g, 49.2 mmol) in dichloromethane (100 ml), and then the reaction mixture was stirred at 25°C for 3 hours. The reaction mixture was concentrated to obtain a crude product, which was purified to obtain a yellow oil compound A1 (11.2 g, 74.2%). MS: 614.2 (M+H) + . 1 H NMR(400MHz, DMSO-d6)δ7.83(d,J=9.2Hz,1H),5.22(d,J=3.2Hz,1H),4.96(dd, J=11.2,3.4Hz,1H),4.51(d,J=8.4Hz,1H),4.03(s,3H),3.90(dd,J=20.0,9.0H z,1H),3.62(dtd,J=15.8,10.2,6.0Hz,2H),2.79(t,J=7.4Hz,2H),2.11(s,3H) ,2.00(s,3H),1.89(s,3H),1.76(s,3H),1.73–1.64(m,2H),1.63–1.54(m,2H).

[0156] Preparation of Example Compound A2 (L96-deAc)

[0157]

[0158]

[0159] Step 1: Preparation of compound A2-1

[0160] Synthesis of Compound A2-1 Referring to Compound A1, Compound A2-1 was prepared by using Compound Int.2 instead of Compound A1-3. MS: 509.1 (M+Na) + .

[0161] Step 2: Preparation of compound A2-2

[0162] Under nitrogen protection, a reaction mixture of compound A3 (400 mg, 0.223 mmol), compound A2-1 (163 mg, 0.334 mmol) and DIEA (144 mg, 1.115 mmol) in acetonitrile (20 ml) was stirred at 20 degrees for 40 hours; the reaction mixture was then concentrated to obtain a crude product, which was purified to obtain a white solid compound A2-2 (270 mg, 57.8%). MS: 1049.3 (1 / 2M+H) + .

[0163] Step 3: Preparation of compound A2-3

[0164] Compound A2-2 (270 mg, 0.129 mmol) was dissolved in tetrahydrofuran (10 ml), palladium carbon (14 mg) was added to the reaction mixture under stirring, and then the reaction mixture was stirred under hydrogen pressure at room temperature for 16 hours; the reaction mixture was filtered and concentrated to obtain compound A2-3 (210 mg, 81%). MS: 1003.6 (1 / 2M+H) + .

[0165] Step 4: Preparation of Compound A2-4

[0166] Under nitrogen protection and stirring at 0 degrees, compound Int.3 (56.8 mg, 0.209 mmol) was slowly added to a solution of compound A2-3 (210 mg, 0.105 mmol) and DIEA (67.6 mg, 0.523 mmol) in acetonitrile (10 ml), and then the reaction mixture was stirred at 20°C for 15 minutes. The reaction mixture was then diluted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound A2-4 (220 mg, 97%). MS: 1087.2 (1 / 2M+H) + .

[0167] Step 5: Preparation of Compound A2-5

[0168] The reaction mixture of compound A2-4 (220 mg, 0.101 mmol), compound Int.7 (63.7 mg, 0.152 mmol) and DIEA (66 mg, 0.506 mmol) in acetonitrile (10 ml) was stirred at 20 degrees for 15 minutes; then the reaction mixture was diluted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was separated and purified to obtain compound A2-5 (120 mg, 49.2%). MS: 1053.7 (1 / 2 [M-DMTr] + H) + .

[0169] Step 6: Preparation of Compound A2

[0170] The reaction mixture of compound A2-5 (120 mg, 0.05 mmol) and 7M ammonia in methanol (6 ml) was stirred at 20 degrees for 4 hours; the reaction mixture was then concentrated to obtain a crude product, which was separated and purified to obtain compound A2 (30 mg, 29.7%). MS: 864.7 (1 / 2 [M-DMTr] + H) + . 1 H NMR (400MHz, DMSO-d6) δ7.88(t,J=5.7Hz,3H),7.78(t,J=5.7Hz,3H),7.66( d,J=9.0Hz,3H),7.36–7.29(m,4H),7.21(td,J=8.8,3.0Hz,5H),7.03(s,1H) ,6.91–6.87(m,4H),5.01(d,J=4.0Hz,1H),4.65–4.56(m,6H),4.51(d,J=4. 3Hz,3H),4.23(d,J=8.4Hz,3H),4.15(d,J=11.9Hz,1H),3.75(s,6H),3.73–3 .63(m,9H),3.54(td,J=11.7,11.3,5.6Hz,21H),3.44(ddd,J=9.9,6.2,2.9 Hz,3H),3.31(q,J=6.2,5.1Hz,4H),3.17(s,1H),3.05(p,J=6.4Hz,15H),2.3 0(t,J=6.4Hz,6H),2.22(t,J=7.4Hz,2H),2.05(q,J=11.3,9.2Hz,10H),1.82 (s, 9H), 1.52 (dd, J = 12.7, 6.6 Hz, 16H), 1.43 (t, J = 7.2 Hz, 6H), 1.23 (s, 12H).

[0171] Preparation Example Preparation of Compound A3

[0172]

[0173] Step 1: Preparation of compound A3-1

[0174] Tert-butyl acrylate (1060 g, 8260 mmol) and NaOH (5M, 19.8 ml) were slowly added to the DMSO (100 ml) reaction solution of compound Int.6 (100 g, 825.5 mmol) in sequence. After the addition, the reaction mixture was stirred at 20°C for 28 hours; then the reaction mixture was extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a light yellow oil compound A3-1 (74 g, 17.7%). MS: 506.4 (M+H) + .

[0175] Step 2: Preparation of compound A3-2

[0176] Under stirring conditions at 0 degrees, CbzCl (22.7 g, 133.6 mmol) was slowly added to a dichloromethane (100 ml) reaction solution of compound A3-1 (15 g, 29.7 mmol) and TEA (9.0 g, 89.1 mmol). After the addition, the reaction mixture was stirred at 20°C for 12 hours; then the reaction mixture was diluted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a yellow oil compound A3-2 (18.8 g). 1 H NMR (400MHz, DMSO-d6) δ7.36–7.30(m,6H),4.97(s,2H),3.54(t,J=6.4Hz,6H),3.51(s,6H),2.38(t,J=6.0Hz,6H),1.39(s,27H).

[0177] Step 3: Preparation of compound A3-3

[0178] The reaction mixture of compound A3-2 (18.8 g) and formic acid (200 ml) was stirred at 20°C for 12 hours; then the reaction mixture was concentrated to obtain a yellow oil compound A3-3 (16.5 g). 1H NMR (400 MHz, DMSO) δ12.12 (s, 3H), 7.40–7.32 (m, 5H), 5.17 (s, 1H), 4.99 (s, 2H), 3.57 (t, J=6.4 Hz, 6H), 3.50 (s, 6H), 2.42 (t, J=6.4 Hz, 6H).

[0179] Step 4: Preparation of Compound A3-4

[0180] Under nitrogen protection and stirring at room temperature, HOBt (13.2 g, 97.6 mmol) and EDCI (18.7 g, 97.6 mmol) were slowly added to a DMF (120 ml) solution of compound A3-3 (11.5 g, 24.4 mmol), N-tert-butyloxycarbonyl-1,3-propylenediamine (17.0 g, 97.6 mmol) and DIEA (25 g, 193.6 mmol). After the addition, the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with ethyl acetate and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a yellow oil compound A3-4 (16 g). 1 HNMR (400MHz, CDCl3) δ7.37–7.28(m,5H),6.83(s,3H),5.55(s,1H),5.16(t,J=6.4Hz,3H),5.03(s,2H),3.68(t,J=6. 0Hz, 6H), 3.65 (s, 6H), 3.27–3.24 (m, 6H), 3.14–3.10 (m, 6H), 2.40 (t, J = 5.6Hz, 6H), 1.70–1.47 (m, 6H), 1.43 (s, 27H).

[0181] Step 5: Preparation of Compound A3-5

[0182] A reaction mixture of compound A3-4 (16 g) in dichloromethane (100 ml) and trifluoroacetic acid (20 ml) was stirred at 20°C for 12 hours; the reaction mixture was then concentrated to obtain a yellow oil compound A3-5 (20 g), which was used directly in the next step without purification.

[0183] Step 6: Preparation of Compound A3-6

[0184] Under stirring at 0 degrees and nitrogen protection, compound A1 (5 g) in dichloromethane (20 ml) was slowly added to the reaction mixture of compound A3-5 (5 g) and DIEA (2.8 g) in acetonitrile (20 ml). After the addition, the reaction mixture was stirred at room temperature for 12 hours; then the reaction mixture was diluted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified to give a white solid compound A3-6 (3.85 g). 1H NMR(400MHz,DMSO-d6)δ7.82-7.79(m,6H),7.71(t,J=5.6Hz,3H),7.37-7.27(m,5H),6.51(s,1H),5 .21(d,J=7.6Hz,3H),4.98-4.95(m,5H),4.48(d,J=7.6Hz,3H),4.04-4.00(m,9H),3.90-3.83(m,3H ),3.72-3.67(m,3H),3.56-3.48(m,12H),3.43-3.37(m,3H),3.05-2.99(m,12H),2.27(t,J=6.4Hz, 6H), 2.10 (s, 9H), 2.04 (t, J = 7.6Hz, 6H), 1.99 (s, 9H), 1.89 (s, 9H), 1.77 (s, 9H), 1.51–1.45 (m, 18H).

[0185] Step 7: Preparation of Compound A3

[0186] Compound A3-6 (4.5 g) was dissolved in methanol (25 ml), palladium carbon (1 g) was added to the reaction mixture under stirring, and then the reaction mixture was stirred under hydrogen pressure at room temperature for 1.5 hours; the reaction mixture was filtered and concentrated to obtain white solid compound A3 (4.2 g).

[0187] Preparation Example Preparation of Compound A4

[0188]

[0189] The synthesis of compound A4 was prepared by referring to compound A1, using compound A1-1 as the starting material to obtain compound A4. MS: 628.4 (M+H) + .

[0190] Preparation Example Preparation of Compound A5

[0191]

[0192] The synthesis of compound A5 was prepared by referring to compound A7-1, using compound A1-1 as the starting material to obtain compound A5. MS: 419.4 (M+H) + .

[0193] Preparation Example Preparation of Compound A6

[0194]

[0195] The synthesis of compound A6 was prepared by referring to compound A8, using compound A1-1 as the starting material to obtain compound A6. MS: 479.4 (M+H) + .

[0196] Preparation Example Preparation of Compound A7

[0197]

[0198] The synthesis of compound A7-1 was prepared by referring to compound A8 using compound A1-1 as the starting material. MS: 552.4 (M+H) + .

[0199] The synthesis of compound A7 was prepared by referring to compound A1, and compound A7-1 was used instead of compound A1-3 to obtain compound A7. MS: 718.1 (M+H) + .

[0200] Preparation Example Preparation of Compound A8

[0201]

[0202] The synthesis of compound A8-1 was prepared by referring to compound A1-2, and compound A8-1 was prepared by using compound A1-1 as a starting material. MS: 549.6 (M+H) + .

[0203] The synthesis of compound A8 was prepared by referring to compound A3, and compound A8-1 was used instead of compound A3-6 to obtain compound A8. MS: 523.4 (M+H) + .

[0204] Preparation Example Preparation of Compound A14-4

[0205]

[0206] The synthesis of compound A14-4 was prepared by referring to compound A1, using compound A1-1 as the starting material to obtain compound A14-4. MS: 670.4 (M+H) + .

[0207] Preparation of Example Compound A10

[0208]

[0209] Step 1: Preparation of compound A10-1

[0210] Under stirring conditions at room temperature, saturated sodium bicarbonate solution (30 ml) and CbzCl (5.4 g, 32 mmol) were added to a solution of (3-aminopropyl)carbamic acid tert-butyl ester (5 g, 29 mmol) in dichloromethane (10 ml). After the addition, the reaction mixture was stirred at 20°C for 1 hour; then the reaction mixture was extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a white solid (3-tert-butoxycarbonylamino-propyl)-carbamic acid benzyl ester (7 g). MS: 309.4 (M+H) + .

[0211] Under stirring conditions at 0 degrees, trifluoroacetic acid (10 ml) was slowly added to a solution of (3-tert-butoxycarbonylamino-propyl)-carbamic acid benzyl ester (3 g, 9.7 mmol) in dichloromethane (10 ml), and then the reaction mixture was stirred at 25°C for 3 hours; then the reaction mixture was concentrated to obtain compound A10-1 (5 g) trifluoroacetate. MS: 209.2 (M+H) + .

[0212] Step 2: Preparation of compound A10-2

[0213] Under stirring conditions at 20 degrees, TEA (5.8 g, 58 mmol) and glutaric anhydride (1.25 g, 12.5 mmol) were slowly added to the dichloromethane (10 ml) reaction solution of the trifluoroacetate salt of compound A10-1 (5 g, 9.6 mmol). After the addition, the reaction mixture was stirred at 20°C for 16 hours; then the reaction mixture was diluted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a white solid compound A10-2 (2.5 g). MS: 323.2 (M+H) + .

[0214] Step 3: Preparation of compound A10-3

[0215] Under stirring conditions at 20 degrees, TEA (2.35 g, 23.3 mmol) and intermediate Int.3 (2.6 g, 9.3 mmol) were slowly added to the dichloromethane (10 ml) reaction solution of compound A10-2 (2.5 g, 7.76 mmol). After the addition, the reaction mixture was stirred at 20°C for 2 hours; then the reaction mixture was diluted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a white solid compound A10-3 (2.5 g). MS: 489.2 (M+H) + .

[0216] Step 4: Preparation of compound A10-4

[0217] Under nitrogen protection, a reaction mixture of compound A3 (1 g, 0.558 mmol), compound A10-3 (410 mg, 0.839 mmol) and TEA (0.17 g, 1.67 mmol) in DMF (5 ml) was stirred at 35 degrees for 16 hours; the reaction mixture was then diluted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a white solid compound A10-4 (0.9 g). MS: 1096.7 (1 / 2M+H) + .

[0218] Step 5: Preparation of Compound A10-5

[0219] Compound A10-4 (900 mg, 0.43 mmol) was dissolved in methanol (20 ml), palladium carbon (600 mg) was added to the reaction mixture under stirring, and then the reaction mixture was stirred under hydrogen pressure at room temperature for 0.5 hours; the reaction mixture was filtered and concentrated to obtain compound A10-5 (750 mg), which was directly used in the next step without purification. MS: 982.8 (1 / 2M+H) + .

[0220] Step 6: Preparation of Compound A10-6

[0221] Under nitrogen protection at 20 degrees, TEA (93 mg, 0.93 mmol) and Cy5-NHS-ester (100 mg, 0.135 mmol) were slowly added to the DMF (2 ml) reaction mixture of compound A10-5 (344 mg, 0.176 mmol). After the addition, the reaction mixture was stirred at room temperature for 16 hours; then the reaction mixture was diluted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product compound A10-6 (120 mg). MS: 1295.0 (1 / 2M+H) + .

[0222] Step 7: Preparation of Compound A10

[0223] The reaction mixture of compound A10-6 (120 mg, 0.046 mmol) and 7M ammonia in methanol (6 ml) was stirred at 20 degrees for 2 hours; the reaction mixture was then concentrated to obtain a crude product, which was separated and purified to obtain compound A10 (22.1 mg). MS: 1105.9 (1 / 2 [M] + H) + . 1H NMR (400MHz, DMSO-d6) δ8.33(t,J=13.2Hz,2H),7.79(s,2H),7.67(d,J=5.1Hz,2H),7.33(dd,J=8.3, 2.7Hz,2H),6.59(t,J=12.3Hz,1H),6.29(t,J=14.2Hz,2H),4.23(d,J=8.4Hz,3H),4.09(s,2H),3.73–

[0224] 3.66(m,6H),3.65(d,J=3.0Hz,3H),3.60(s,3H),3.54(d,J=2.9Hz,6H),3.49( dd,J=11.1,6.4Hz,11H),3.42(dd,J=10.5,3.1Hz,3H),3.38–3.33(m,3H),3.31 (t,J=6.3Hz,3H),3.06–2.96(m,17H),2.28(t,J=6.1Hz,6H),2.05(t,J=7.2Hz ,12H),1.81(s,9H),1.69(s,12H),1.56–1.37(m,24H),1.28(d,J=26.7Hz,4H).

[0225] Preparation of Example Compound A11

[0226]

[0227] Step 1: Preparation of Compound A11-1 A reaction mixture of sodium bicarbonate (0.837 g, 9.96 mmol), CbzCl (1.87 g, 10.96 mmol) and 4-(2-aminoethyl)phenol hydrochloride (1.73 g, 9.96 mmol) in tetrahydrofuran (80 ml) and water (80 ml) was stirred at 20°C for 16 hours; the reaction mixture was then poured into ice water and the pH value was adjusted to 4 with 5% citric acid, then diluted and extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a yellow solid compound A11-1 (2 g, 74%). MS: 273.2 (M+H) + .

[0228] Step 2: Preparation of Compound A11-2 Potassium carbonate (3.06 g, 22.1 mmol), (3-bromopropyl) carbamic acid tert-butyl ester (2.282 g, 9.58 mmol) and Compound A11-1 (2 g, 7.37 mmol) in DMF (80 ml) were stirred at 70°C for 4 hours; the reaction mixture was then poured into ice water and the pH value was adjusted to 4 with 5% citric acid, then diluted and extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a yellow solid Compound A11-2 (2 g, 63%). MS: 429.5 (M+H) + .

[0229] Step 3: Preparation of Compound A11-5

[0230] Synthesis of Compound A11-5 Referring to Compound A3, Compound A11-5 was prepared by using Compound A11-2 instead of Compound A3-4.

[0231] Step 4: Preparation of Compound A11-6

[0232] Synthesis of Compound A11-6 Referring to Compound A11-2, Compound A11-6 was prepared. MS: 474.5 (M+H) + .

[0233] Step 5: Preparation of Compound A11-7

[0234] Synthesis of Compound A11-7 Referring to Compound A11-2, Compound A11-7 was prepared by using Compound A11-6 instead of Compound A1-3. MS: 639.5 (M+H) + .

[0235] Step 6: Preparation of Compound A11-9

[0236] Synthesis of Compound A11-9 Referring to Compound A11-5, Compound A11-9 was prepared by using Compound A11-7 instead of Compound A1. MS: 659.5 (M+H) + .

[0237] Step 7: Preparation of Compound A11-10

[0238] Synthesis of Compound A11-10 Referring to Compound A2-2, Compound A11-10 was prepared by using Compound A11-9 instead of Compound A3. MS: 659.5 (M+H) + .

[0239]

[0240]

[0241] Step 8: Preparation of Compound A11-11

[0242] Synthesis of Compound A11-11 Referring to Compound A11-4, Compound A11-11 was prepared by using Compound A11-10 instead of Compound A11-2. MS: 1620.2 (M+H) + .

[0243] Step 9: Preparation of Compound A11-14

[0244] Synthesis of Compound A11-14 Referring to Compound A11-9, Compound A11-11 was used instead of Compound A11-6 to prepare Compound A11-14. MS: 2136.2 (M+H) + .

[0245] Step 10: Preparation of Compound A11

[0246] Synthesis of Compound A11 Referring to Compound A12, Compound A11 was prepared by using Compound A11-14 instead of Compound A12-7. MS: 2159.2 (M+H) + . 1H NMR (400MHz, DMSO-d6) δ8.07(s,1H),7.94(d,J=7.5Hz,2H),7.86(d,J=5.9Hz ,3H),7.65(d,J=9.0Hz,3H),7.33(dd,J=8.1,6.1Hz,4H),7.21(dd,J=8.8,2.7 Hz,5H),7.12(t,J=9.0Hz,5H),7.03(d,J=8.2Hz,1H),6.94–6.75(m,9H),5.0 4–4.88(m,1H),4.67–4.56(m,6H),4.50(d,J=4.3Hz,3H),4.41(d,J=9.1Hz,2H ),4.24(d,J=8.4Hz,3H),4.19–4.09(m,1H),3.93(d,J=6.3Hz,6H),3.81–3.6 3(m,16H),3.63–3.40(m,10H),3.35–3.28(m,10H),3.19(d,J=6.8Hz,8H),3.0 0(d,J=8.5Hz,2H),2.89(d,J=12.3Hz,1H),2.76–2.57(m,3H),2.22(t,J=7.3 Hz,1H),2.12–1.96(m,10H),1.82(s,17H),1.62–1.40(m,15H),1.17(s,10H).

[0247] Preparation of Example Compound A12

[0248]

[0249]

[0250]

[0251] Step 1: Preparation of compound A12-1

[0252] A DMF (10 ml) solution of intermediate Int.12 (3 g, 6.359 mmol), TEA (3.21 g, 31.7 mmol), HATU (7.24 g, 19.04 mmol) and intermediate Int.2 (2.24 g, 7.62 mmol) was stirred at 20°C for 1 hour. The reaction mixture was then poured into ice water and diluted with dichloromethane for extraction. The organic phase was washed with 5% citric acid solution and saturated brine in turn, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow oily compound A12-1 (4.8 g, 97.5%). MS: 775.7 (M+H) + .

[0253] Step 2: Preparation of compound A12-2

[0254] A solution of trifluoroacetic acid (4.41 g, 38.7 mmol) and compound A12-1 (1 g, 1.29 mmol) in dichloromethane (10 ml) was stirred at 20°C for 1 hour. The reaction mixture was then poured into ice water and diluted with dichloromethane for extraction. The organic phase was washed with 5% citric acid solution and saturated brine in sequence, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified to obtain yellow oily compound A12-2 (0.6 g, 98%). MS: 475.6 (M+H) + .

[0255] Step 3: Preparation of compound A12-3

[0256] A reaction solution of TEA (6.4 g, 63.2 mmol) and glutaric anhydride (4.33 g, 37.9 mmol) and compound A12-2 (5 g, 9.6 mmol) in dichloroethane (10 ml) was stirred at 50°C for 16 hours; the reaction mixture was then concentrated to obtain a crude product, which was purified to obtain a yellow oily compound A12-3 (8.5 g, 99%). MS: 817.7 (M+H) + .

[0257] Step 4: Preparation of Compound A12-4 A DMF (10 ml) reaction solution of (3-aminopropyl)carbamic acid tert-butyl ester (3.07 g, 17.63 mmol), DIEA (1.898 g, 14.69 mmol), Compound A12-3 (4 g, 2.94 mmol) and HATU (4.47 g, 11.75 mmol) was stirred at 20°C for 40 hours; the reaction mixture was then concentrated to obtain a crude product, which was purified to obtain a yellow oily compound A12-4 (630 mg, 16.7%). MS: 1286.1 (M+H) + .

[0258] Step 5: Preparation of Compound A12-5

[0259] A solution of trifluoroacetic acid (1.676 g, 14.7 mmol) and compound A12-4 (630 mg, 0.49 mmol) in dichloromethane (10 ml) was stirred at 20°C for 40 hours. The reaction mixture was then concentrated to obtain a crude product, which was purified to obtain a yellow oily compound A12-5 (0.48 g, 99.9%). MS: 329.6 (M / 3+H) + .

[0260] Step 6: Preparation of Compound A12-6

[0261] The reaction mixture of compound A1 (1.195 g, 1.949 mmol), DIEA (630 mg, 4.87 mmol) and compound A12-5 (480 mg, 0.487 mmol) in dichloromethane (10 ml) was stirred at room temperature for 40 hours; the reaction mixture was then filtered and concentrated to obtain a crude product, which was purified to obtain a white solid compound A12-6 (0.77 g, 69.5%). MS: 1138.0 (M / 2+H) + .

[0262] Step 7: Preparation of Compound A12-7

[0263] Compound A12-6 (600 mg, 0.264 mmol) was dissolved in tetrahydrofuran (20 ml), palladium carbon (28 mg) was added to the reaction mixture under stirring, and then the reaction mixture was stirred under hydrogen pressure at room temperature for 40 hours; the reaction mixture was filtered and concentrated to obtain a crude product, which was purified to obtain a white solid compound A12-7 (0.52 g, 90%). MS: 1092.9 (M / 2+H) + .

[0264] Step 8: Preparation of Compound A12-8

[0265] Under nitrogen protection at 0 degrees, compound Int.3 (133 mg, 0.476 mmol) was slowly added to a solution of compound A12-7 (520 mg, 0.238 mmol) and DIEA (154 mg, 1.19 mmol) in acetonitrile (10 ml), and the reaction mixture was stirred at 20°C for 0.25 hours. The reaction mixture was then diluted and extracted with dichloromethane, and the organic phase was washed with 5% citric acid solution and saturated brine in turn, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude yellow oil compound A12-8 (560 mg, 100%). MS: 1175.9 (M / 2+H) + .

[0266] Step 9: Preparation of Compound A12-9

[0267] The reaction mixture of compound A12-8 (560 mg, 0.238 mmol), compound Int.7 (150 mg, 0.358 mmol) and DIEA (154 mg, 1.192 mmol) in acetonitrile (10 ml) was stirred at 20 degrees for 15 minutes; the reaction mixture was then concentrated to obtain a crude product, which was separated and purified to obtain compound A12-9 (310 mg, 50.3%). MS: 1142.4 (1 / 2 [M-DMTr] + H) + .

[0268] Step 10: Preparation of Compound A12

[0269] The reaction mixture of compound A12-9 (110 mg, 0.043 mmol) and 7M ammonia in methanol (10 ml) was stirred at 20 degrees for 4 hours; the reaction mixture was then concentrated to obtain a crude product, which was separated and purified to obtain a white solid compound A12 (45 mg, 47.9%). MS: 953.2 (1 / 2 [M-DMTr] + H) + . 1 H NMR (400MHz, DMSO-d6) δ7.81(d,J=18.1Hz,6H),7.68(d,J=9.1Hz,3H),7.36–7.30(m,4H),7.22(ddd,J=8.8,6.7,2.6Hz,5H),6.90(ddd,J=8.9,6.0,2 .4Hz,4H),5.04(d,J=4.1Hz,1H),4.65(t,J=5.7Hz,3H),4.60(d,J=6.3Hz, 3H), 4.52 (d, J = 4.4Hz, 3H), 4.24 (d, J = 8.4Hz, 3H), 4.16 (d, J = 9.4Hz, 1H), 3. 77–3.66(m,16H),3.54(tt,J=11.1,5.3Hz,22H),3.34(dt,J=21.3,5.4Hz, 12H),3.19(dd,J=9.0,5.0Hz,1H),3.08–2.99(m,14H),2.35(s,8H),2.23(t ,J=7.5Hz,2H),2.13(s,6H),2.06(d,J=7.2Hz,6H),1.82(s,9H),1.76(d,J =9.0Hz, 6H), 1.53 (d, J = 11.9Hz, 16H), 1.45 (d, J = 7.1Hz, 6H), 1.26 (s, 12H).

[0270] Preparation of Example Compound A14

[0271]

[0272] Step 1: Preparation of compound A14-1

[0273] Synthesis of Compound A14-1 Referring to Compound A12-9, Compound A14-1 was prepared by using (S)-2,3-bis((tert-butyloxycarbonyl)amino)propionic acid. MS: 491.4 (M+H) + .

[0274] Step 2: Preparation of Compound A14-5

[0275] Synthesis of Compound A14 Referring to Compound A12, Compound A14 was prepared by using Compound A14-1 instead of Compound A12-3. MS: 776.6 (1 / 2 [M-DMTr] + H) + . 1 H NMR (400MHz, DMSO-d6) δ8.22(d,J=7.6Hz,0H),8.03(d,J=6.0Hz,0H),7.92(d,J=6.0Hz,1H),7.83(t,J=5.9Hz,1H),7.63(d,J= 9.0Hz, 4H), 7.32 (q, J=7.7Hz, 4H), 7.22 (td, J=8.8, 2.9Hz, 5H), 6.90 (dt, J=8.5, 3.6Hz, 4H), 4.96 (dd, J=31.0, 4.1Hz, 1H), 4.6 9–4.53(m,6H),4.53–4.38(m,3H),4.38-4.08(m,5H),3.86–3.64(m,16H),3.64-3.42(m,12H),3.34–3.15(m,7H),3.13-2.93( m,3H),2.30-2.14(m,3H),2.13-1.92(m,5H),1.87(d,J=7.9Hz,1H),1.82(s,9H),1.60-1.35(m,17H),1.25(d,J=6.8Hz,41H).

[0276] Preparation of Example Compound A16

[0277]

[0278] Step 1: Preparation of compound A16-1

[0279] Under stirring at 40 degrees, thionyl chloride (15.24 g, 128 mmol) was slowly added to a dichloromethane (20 ml) reaction solution of adamantane-1,3,5,7-tetracarboxylic acid (2 g, 6.4 mmol) and DMF (0.468 g, 6.4 mmol). After the addition, the reaction mixture was stirred at 40°C for 40 hours. The reaction mixture was then concentrated to obtain a crude product, compound A16-1 (2.47 g, 99%). MS: 369.2 (M+H) + .

[0280] Step 2: Preparation of compound A16-2

[0281] A reaction solution of 11-aminoundecanoic acid benzyl ester hydrochloride (2.429 g, 7.04 mmol) and compound A16-1 (0.92 g, 2.383 mmol) in dichloromethane (20 ml) was stirred at 20°C for 16 hours. The reaction mixture was then concentrated to obtain a crude product, which was purified to obtain compound A16-2 (310 mg, 22%). MS: 585.5 (M+H) + .

[0282] Step 3: Preparation of Compound A16

[0283] Synthesis of Compound A16 Referring to Compound A12, Compound A16 was prepared by using Compound A16-2 instead of Compound A12-3. MS: 837.5 (1 / 2[M-DMTr]+H) + . 1 H NMR (500MHz, DMSO-d6) δ7.76(t,J=5.7Hz,3H),7.64(d,J=9.1Hz,3H),7.56(t,J=5.8Hz,3H),7.51(t,J=5.3Hz,1H),7.30(t,J=8.8Hz,4H),7.19(ddd ,J=12.0,8.5,3.5Hz,5H),6.87(dt,J=8.6,4.7Hz,4H),4.95(d,J=43.3Hz ,4H),4.60(s,6H),4.49(s,3H),4.38(s,1H),4.34–4.26(m,1H),4.21(d,J =8.4Hz,3H),4.12(d,J=15.6Hz,2H),3.83–3.61(m,16H),3.60–3.42(m,1 4H),3.28(t,J=6.1Hz,6H),3.16(dd,J=8.8,4.9Hz,2H),3.02(dq,J=20.2, 6.5Hz,15H),2.21(d,J=7.4Hz,2H),2.04(t,J=7.4Hz,7H),1.93(t,J=9.2H z,1H),1.79(s,9H),1.72(s,12H),1.56-1.32(m,21H),1.29–1.06(m,8H).

[0284] Preparation of Example Compound A17

[0285]

[0286] The synthesis of compound A17 was prepared by referring to compound A12, and compound A16-2 was used instead of compound A12-3 to obtain compound A17. MS: 866.1 (1 / 2 [M-DMTr] + H)+ . 1 H NMR (400MHz, DMSO-d6) δ7.60(d,J=9.0Hz,2H),7.55(t,J=5.8Hz,3H),7.48(s,1H),7.35–7.25(m,4H),7.19(td,J=8.8,2 .7Hz,5H),6.87(ddd,J=8.9,5.9,2.5Hz,4H),4.50(s,3H),4.38(d,J=6.2Hz,1H),4.28(d,J=8.4Hz,2H),4.15(d,J=8.2H z,1H),3.81–3.66(m,12H),3.64(d,J=3.0Hz,3H),3.61–3.45(m,46H),3.45–3.37(m,20H),3.23-3.23(m,12H),3.07-2. 93(m,5H),2.25-2.15(m,1H),2.01(dt,J=12.1,5.4Hz,1H),1.86(s,1H),1.80(s,8H),1.71(s,12H),1.52-1.12(m,15H).

[0287] Preparation of Example Compound A18

[0288]

[0289]

[0290] Step 1: Preparation of compound A18-1

[0291] A solution of compound A2-1 (3.01 g, 6.19 mmol), 1,4,7-triazolidine (200 mg, 1.55 mmol) and DIEA (800 mg, 6.19 mmol) in acetonitrile (20 ml) was stirred at 20 degrees for 12 hours. The reaction mixture was poured into ice water, and the pH value was adjusted to 4 with 5% citric acid solution, and then extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a white oily compound A18-1 (1.2 g, 74.8%). MS: 1037.2 (M+H) + .

[0292] Step 2: Preparation of compound A18-2

[0293] Potassium hydroxide (0.13 g, 2.316 mmol) was added to a tetrahydrofuran (5 ml) solution of compound A18-1 (1.2 g, 1.158 mmol) and benzyl alcohol (3.16 g, 29.2 mmol); the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was diluted and extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain a yellow oil compound A18-2 (310 mg, 74.8%). MS: 857.5 (M+H) + .

[0294] Step 3: Preparation of Compound A18

[0295] Synthesis of Compound A18 Referring to Compound A12, Compound A18 was prepared by using Compound A18-2. MS: 1831.3 (1 / 2 [M-DMTr] + H) + . 1 H NMR (400MHz, DMSO-d6) δ7.86(t,J=5.2Hz,5H),7.76(t,J=5.4Hz,5H),7.65(d,J=9.0Hz,5H),7.35–7.31(m,3H),7.2 4–7.20(m,4H),7.02(s,1H),6.91–6.87(m,4H),5.06–4.96(m,1H),4.94–4.86(m,1H),4.62–4.56(m,12H),4.50(d,J =4.3Hz,5H),4.24(d,J=8.4Hz,5H),3.77–3.66(m,25H),3.62–3.42(m,52H),3.40–3.36(m,7H),3.33–3.23(m,14H), 3.12–3.00(m,27H),2.34–2.23(m,18H),2.11–2.01(m,18H),1.82(s,18H),1.59–1.39(m,51H),1.32–1.19(m,37H).

[0296] Preparation of Example Compound A19

[0297]

[0298]

[0299] Step 1: Preparation of compound A19-2

[0300] The synthesis of compound A19-2 was prepared by referring to compound A3 by using 2-amino-2-methylpropane-1,3-diol instead of 2-amino-2-(hydroxymethyl)propane-1,3-diol. MS: 611.3 (M / 2+H) +

[0301] Step 2: Preparation of Compound A19

[0302] Synthesis of Compound A19 Referring to Compound A18, Compound A19 was prepared by using Compound A19-2 instead of Compound A3. MS: 923.3 (1 / 3 [M-DMTr] + H) + . 1 H NMR (400MHz, DMSO-d6) δ7.86(t,J=5.7Hz,2H),7.76(t,J=5.6Hz,4H),7.65(d,J=9.1Hz,4H),7.38–7.31(m,6H),7.27–7 .19(m,5H),7.13(s,1H),6.93–6.86(m,4H),5.04(s,1H),4.60(s,6H),4.50(s,4H),4.25(d,J=8.4Hz,4H),3.77–3.65(m ,18H),3.61–3.42(m,53H),3.18(t,J=6.1Hz,4H),3.06(d,J=7.7Hz,16H),2.70(p,J=1.9Hz,2H),2.38–2.34(m,2H),2.3 5–2.20(m,16H),2.06(q,J=8.4,7.8Hz,12H),1.82(s,12H),1.51(dd,J=20.6,13.4Hz,34H),1.25(s,36H),1.18(s,6H).

[0303] Preparation of Example Compound A20

[0304]

[0305] Step 1: Preparation of compound A20-1

[0306] Under nitrogen protection at -78 degrees, ethyl trifluoroacetate (19.43 g, 137 mmol) was slowly added dropwise to a methanol (300 ml) solution of N, N'-bis (2-aminoethyl) ethylenediamine (20 g, 137 mmol); after the addition, the reaction mixture was stirred and reacted for 45 minutes at -78 degrees, and the reaction solution was heated to 0 degrees and continued to react for 1 hour. A methanol (30 ml) solution of di-tert-butyl dicarbonate (112 g, 513 mmol) was slowly added dropwise to the above reaction solution under stirring at 0 degrees; and the reaction mixture was stirred and reacted at 20°C for 20 hours. Then the reaction mixture was concentrated to obtain a crude product, which was dissolved in dichloromethane, and the organic phase was washed with 10% sodium bicarbonate solution, washed with saturated brine, dried with anhydrous sodium sulfate, filtered, concentrated, and then treated with methyl tert-butyl ether to precipitate a white solid, which was filtered to obtain a white solid compound A20-1 (32 g, 43%). MS:543.1(M+H) + .

[0307] Step 2: Preparation of compound A20-2

[0308] A mixture of compound A20-2 (14 g, 25.8 mmol), potassium carbonate (7.13 g, 51.6 mmol) in methanol (50 ml) and water (50 ml) was refluxed and stirred for 4 hours; the reaction mixture was then concentrated and diluted with dichloromethane for extraction, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was separated and purified to obtain compound A20-2 (3.5 g, 30%). MS: 447.1 (M+H) + . 1 H NMR (400MHz, DMSO-d6) δ6.85 (s, 1H), 3.26 (s, 4H), 3.16 (d, J = 6.6Hz, 2H), 3. 11(s,2H),3.05(d,J=5.9Hz,2H),2.64(t,J=6.9Hz,2H),1.47–1.33(m,27H).

[0309] Step 3: Preparation of Compound A20-5

[0310] Synthesis of Compound A20-5 Reference Compound A12-2 was used to prepare Compound A20-5. MS: 789.1 (M+H) + .

[0311] Step 4: Preparation of Compound A20

[0312] The synthesis of compound A20 was prepared by referring to compound A12, and compound A20-5 was used instead of compound A12-5 to obtain compound A20. MS: 854.6 (1 / 2 [M-DMTr] + H) + . 1 H NMR (400MHz, DMSO-d6) δ7.98(s,1H),7.75(d,J=5.4Hz,3H),7.65(d,J=9.0Hz,3H),7.33(t,J=4.5Hz,4H),7.21( dd,J=8.8,2.6Hz,5H),6.94–6.83(m,4H),4.96(dd,J=32.3,4.0Hz,1H),4.64–4.55(m,6H),4.50(d,J=4.2Hz,3H) ,4.25(d,J=8.4Hz,4H),3.80–3.65(m,16H),3.63–3.42(m,12H),3.32(t,J=6.3Hz,10H),3.24–3.11(m,5H),3.0 2(s,8H),2.23(s,6H),2.06(t,J=7.4Hz,12H),1.83(s,9H),1.45(ddt,J=36.3,14.4,7.4Hz,31H),1.24(s,18H).

[0313] Preparation of Example Compound A23

[0314]

[0315] The synthesis of compound A23 was prepared by referring to compound A12, and compound A12-3 was used as the starting material to obtain compound A23. MS: 825.6 (1 / 2 [M-DMTr] + H) + . 1 H NMR (400MHz, DMSO-d6) δ7.77(s,3H),7.63(d,J=9.5Hz,3H),7.33(d,J=6.7Hz,4H),7.20(t,J=4 .8Hz,5H),6.89(s,4H),5.00(s,1H),4.64–4.54(m,6H),4.48(d,J=4.3Hz,3H),4.23(d,J=8.4H z,3H),4.22(s,1H),3.70(d,J=36.7Hz,18H),3.53(d,J=6.6Hz,25H),3.03(d,J=6.7Hz,12H),2 .34(s,10H),2.22(s,2H),2.11(s,6H),1.81(s,9H),1.73(s,6H),1.43(s,18H),1.25(s,12H).

[0316] Preparation of Example Compound A24

[0317]

[0318] The synthesis of compound A24 was prepared by referring to compound A12, and compound A12-3 was used as the starting material to obtain compound A24. MS: 915.7 (1 / 2 [M-DMTr] + H) + . 1 H NMR (400MHz, DMSO-d6) δ7.89(d,J=6.3Hz,1H),7.63(d,J=9.0Hz,3H),7.32(t,J=7.0Hz,4H),7.21(td,J=8.9,3.2Hz, 5H),6.91–6.86(m,4H),5.01(s,1H),4.93(s,1H),4.62(s,3H),4.58(d,J=6.4Hz,4H),4.53–4.48(m,4H),4.41(s,1H) ,4.30(d,J=8.4Hz,4H),4.17(d,J=9.3Hz,2H),3.82–3.64(m,25H),3.54–3.48(m,45H),3.20(q,J=6.0Hz,6H),3.04– 2.96(m,2H),2.43–1.98(m,16H),1.87(t,J=13.9Hz,2H),1.82(s,9H),1.78–1.68(m,6H),1.49(s,3H),1.25(s,12H).

[0319] Preparation of Example Compound A25

[0320]

[0321] The synthesis of compound A25 was prepared by referring to compound A12 and using compound A16-2 as the starting material. MS: 879.7 (1 / 2 [M-DMTr] + H) + . 1H NMR (400MHz, DMSO-d6) δ7.71(t,J=5.7Hz,3H),7.63(d,J=9.1Hz,3H),7.49(d,J=6.1Hz,4H),7.32(q,J=7.8,7.4Hz,4H),7.26–7. 17(m,5H),6.93–6.84(m,4H),4.99–4.90(m,4H),4.62–4.53(m,6H),4.47(d,J=4.3Hz,3H),4.24(d,J=8.4Hz,3H),4.16–4.14(m,1 H),3.75(s,6H),3.73–3.65(m,8H),3.54(m,7H),3.48–3.41(m,4H),3.39–3.37(m,4H),3.31–3.29(m,5H),3.18–3.11(m,1H),3.0 2–3.01(m,16H),2.24–2.20(m,1H),2.07–2.03(m,7H),1.82(s,9H),1.73–1.70(m,11H),1.51–1.37(m,29H),1.24–1.19(m,18H).

[0322] Preparation of Example Compound A27

[0323]

[0324] The synthesis of compound A27 was prepared by referring to compound A18. MS: 1008.6 (1 / 2 [M-DMTr] + H) + . 1H NMR (400MHz, DMSO-d6) δ8.06(d,J=8.0Hz,1H),7.61(d,J=9.2Hz,1H),7.29(t,J=7.7Hz,3H),7.24–7.09(m,12H),6.96(t, J=7.3Hz,1H),6.90-6.80(m,4H),5.00-4.85(m,1H),4.65–4.50(m,4H),4.47(d,J=4.3Hz,3H),4.44-4.31(m,4H),4.24(d ,J=8.4Hz,1H),4.17-3.96(m,2H),3.83–3.67(m,12H),3.64(s,2H),3.52(tt,J=10.3,4.9Hz,3H),3.11-2.92(m,12H),2. 2-2.05(m,7H),2.05-1.90(m,7H),1.78(s,12H),1.75(s,3H),1.72–1.48(m,28H),1.47-1.33(m,2H),1.29-1.06(m,14H).

[0325] Preparation of Example Compound A28

[0326]

[0327] Synthesis of Compound A28-1 Referring to Compound A8, Compound A28-1 was prepared by using Compound A1-1 as a starting material.

[0328] The synthesis of compound A28 was prepared by referring to compound A27. MS: 932.2 (1 / 2 [M-DMTr] + H) + . 1H NMR (400MHz, DMSO-d6) δ7.66–7.54(m,6H),7.52–7.49(m,1H),7.36–7.28(m,4H),7.25–7.19(m,5H),6.92–6.87(m,4H),4 .99–4.91(m,1H),4.66–4.54(m,6H),4.51(d,J=4.4Hz,3H),4.42–4.39(m,1H),4.31(d,J=8.4Hz,3H),4.16–4.14(m,1H), 3.85–3.63(m,17H),3.62–3.44(m,55H),3.46–3.39(m,8H),3.32–3.30(m,5H),3.24–3.21(m,7H),3.03–3.29(m,4H),2.2 4–2.20(m,2H),2.04–2.01(m,2H),1.88–1.85(m,2H),1.82(s,9H),1.73(s,11H),1.49–1.39(m,4H),1.24–1.20(m,12H).

[0329] Preparation of Example Compound A29

[0330]

[0331]

[0332] The synthesis of compound A29-2 was prepared by referring to compound A7 using compound A1-1 as the starting material to obtain compound A29-2. MS: 660.4 (M+H) + .

[0333] The synthesis of compound A29 was prepared by referring to compound A27. MS: 767.2 (1 / 2 [M-DMTr] + H) + . 1H NMR(400MHz, DMSO-d6)δ7.61(d,J=9.0Hz,3H),7.36–7.28(m,4H),7.24–7.19(m,5H),6.92–6.87(m,4H),4.98–4.97(m ,1H),4.63–4.52(m,6H),4.48(d,J=4.3Hz,3H),4.42–4.40(m,1H),4.32(d,J=8.5Hz,3H),4.20(s,7H),3.80–3.78(m,4 H),4.76(s,7H),3.75–3.65(m,7H),3.63–3.50(m,42H),3.48–3.43(m,5H),3.33–3.31(m,3H),3.20–3.17(m,1H),3.02 –2.99(m,2H),2.32–2.30(m,2H),2.24–2.21(s,2H),1.88–1.85(m,1H),1.82(s,9H),1.51–1.47(m,4H),1.27(s,12H).

[0334] Preparation of Example Compound A30

[0335]

[0336] The synthesis of compound A30 was prepared by referring to compound A12. MS: 826.2 (1 / 2 [M-DMTr] + H) + . 1 H NMR(400MHz, DMSO-d6)δ7.71(s,4H),7.62(d,J=9.0Hz,3H),7.39–7.34(m,4H),7.24–7.19(m,5H),6.91– 6.87(m,4H),4.60–4.51(m,7H),4.46(d,J=4.3Hz,3H),4.41(s,1H),4.24(d,J=8.4Hz,3H),3.75(s,7H), 3.73–3.64(m,9H),3.63–3.40(m,13H),3.38–3.36(m,4H),3.31–3.29(m,4H),3.21–3.18(m,5H),3.07–2 .94(m,8H),2.30–2.20(m,6H),2.07–2.01(m,10H),1.82(s,9H),1.57–1.33(m,30H),1.25–1.22(m,21H).

[0337] Preparation of Example Compound A32

[0338]

[0339] Synthesis of Compound A32 Compound A32 was prepared by referring to Compound A30. MS: 797.5 (1 / 2 [M-DMTr] + H) + . 1 H NMR(400MHz, DMSO-d6)δ7.70(s,3H),7.62(d,J=9.1Hz,3H),7.53(s,3H),7.42–7.29(m,4H),7.28–7.16(m,5H),6.91– 6.85(m,4H),4.97(s,1H),4.60–4.51(m,3H),4.45(s,2H),4.23(d,J=8.4Hz,3H),3.79–3.62(m,12H),3.60–3.49(m,4 H),3.42(s,15H),3.11–2.86(m,18H),2.70–2.66(m,1H),2.34(d,J=2.1Hz,1H),2.20(d,J=7.1Hz,2H),2.03(d,J=19. 2Hz, 6H), 1.81 (d, J = 2.1Hz, 5H), 1.45 (d, J = 27.2Hz, 12H), 1.37 (s, 9H), 1.25 (d, J = 5.4Hz, 16H), 0.86 (d, J = 7.2Hz, 3H).

[0340] Preparation of Example Compound A35

[0341]

[0342] Synthesis of Compound A35 Compound A35 was prepared by referring to Compound A32. MS: 879.5 (1 / 2 [M-DMTr] + H) + . 1H NMR(400MHz, DMSO-d6)δ7.75(t,J=5.6Hz,3H),7.61–7.59(m,3H),7.51–7.46(m,4H),7.35–7.29(m,4H),7.24–7.19(m,5H),6.91–6.87(m,4H), 4.98–4.97(m,1H),4.57(t,J=5.6Hz,3H),4.52(d,J=6.3Hz,3H),4.45(d,J=4.3Hz,3H),4.42–4.40(m,1H),4.25(d,J=8.4Hz,3H),4.17–4.12(m ,1H),3.75(s,7H),3.73–3.64(m,9H),3.62–3.51(m,8H),3.48–3.42(m ,4H),3.39–3.35(m,4H),3.31–3.26(m,3H),3.20–3.17(m,1H),3.05–3. 01(m,16H),2.04(t,J=7.4Hz,7H),1.88–1.85(m,2H),1.82(s,9H),1.7 3(s,11H),1.54–1.43(m,14H),1.40–1.35(m,15H),1.27–1.23(m,18H).

[0343] Preparation of Example Compound A39

[0344]

[0345] Synthesis of Compound A39-1 Referring to Compound A7-1, Compound A39-1 was prepared by using Compound A1-1 as a starting material.

[0346] The synthesis of compound A39 was prepared by referring to compound A32. MS: 815.5 (1 / 2 [M-DMTr] + H) + . 1H NMR(400MHz, DMSO-d6)δ7.60(d,J=9.0Hz,3H),7.45(t,J=5.5Hz,4H),7.36–7.28(m,4H),7.28–7.18 (m,5H),6.92–6.86(m,4H),4.97(d,J=4.0Hz,1H),4.61–4.49(m,6H),4.44(d,J=4.3Hz,3H),4.25(d ,J=8.4Hz,4H),3.79–3.64(m,15H),3.63–3.41(m,10H),3.40–3.34(m,5H),3.30(d,J=6.0Hz,5H),3 .10–2.97(m,9H),2.28–1.86(m,4H),1.81(s,9H),1.72(s,11H),1.53–1.34(m,16H),1.25(s,42H).

[0347] Preparation of Example Compound A40

[0348]

[0349] Synthesis of Compound A40-1 Compound A40-1 was prepared by referring to Compound A1.

[0350] Compound A40 was prepared by referring to compound A32. MS: 879.8

[0351] (1 / 2[M-DMTr]+H) + . 1H NMR(400MHz, DMSO-d6)δ7.71(t,J=5.6Hz,3H),7.63(d,J=9.0Hz,3H),7.48( d,J=6.2Hz,3H),7.35–7.30(m,4H),7.21(td,J=8.8,2.7Hz,5H),6.89(dt,J =8.8,3.4Hz,4H),4.98(d,J=4.0Hz,1H),4.62–4.53(m,6H),4.47(d,J=4.3H z,3H),4.39(d,J=9.1Hz,1H),4.24(d,J=8.4Hz,3H),4.20–4.09(m,1H),3.75 (s,6H),3.70(d,J=9.3Hz,3H),3.68–3.63(m,6H),3.53(tt,J=10.9,5.2Hz, 8H),3.43(ddd,J=17.1,6.6,3.2Hz,7H),3.02(q,J=7.0Hz,18H),2.72–2.68 (m,1H),2.35(t,J=1.9Hz,1H),2.22(s,2H),2.08(t,J=7.5Hz,6H),1.83(s, 9H), 1.75–1.67 (m, 15H), 1.49 (s, 2H), 1.38 (d, J = 7.1Hz, 14H), 1.24 (s, 27H).

[0352] Preparation of Example Compound A41

[0353]

[0354] The synthesis of compound A41 was prepared by referring to compound A32. MS: 825.9 (1 / 2 [M-DMTr] + H) + . 1H NMR (400MHz, DMSO-d6) δ7.60(d,J=9.0Hz,3H),7.55(s,3H),7.33(t,J=7.3Hz,5H),7.25–7.22(m,3H),7.20(dd,J=8.8,2.5Hz,4H),6.88(dd,J=8 .7,3.3Hz,4H),4.98(d,J=4.0Hz,1H),4.61–4.53(m,6H),4.49(d,J=4.3 Hz,3H),4.41(s,1H),4.31(d,J=8.4Hz,3H),4.16(s,1H),3.79(d,J=5.1 Hz,3H),3.75(s,9H),3.71(t,J=9.3Hz,3H),3.66(t,J=3.8Hz,3H),3.60 –3.49(m,43H),3.45(s,9H),3.21–3.14(m,6H),3.04–2.95(m,6H),2.72 –2.67(m,1H),2.35(s,1H),2.22(t,J=7.5Hz,2H),2.07–1.99(m,2H),1. 82(d,J=1.6Hz,9H),1.49(s,3H),1.37(d,J=18.2Hz,4H),1.24(s,14H).

[0355] Preparation of Example Compound A42

[0356]

[0357] Synthesis of Compound A42-1 Referring to Compound A7-1, Compound A42-1 was prepared by using Compound A1-1 as a starting material.

[0358] The synthesis of compound A42 was prepared by referring to compound A40. MS: 818.2 (1 / 2 [M-DMTr] + H) + . 1H NMR(400MHz, DMSO-d6)δ7.61(d,J=9.1Hz,3H),7.51–7.48(m,4H),7.38–7.29(m,4H),7.23–7.19(m,5 H),6.92–6.86(m,4H),4.61–4.51(m,7H),4.46(d,J=4.3Hz,3H),4.41–4.39(m,1H),4.24(d,J=8.4Hz, 3H),4.21–4.11(m,1H),3.75(s,8H),3.74–3.64(m,9H),3.62–3.41(m,13H),3.39–3.36(m,4H),3.33– 3.26(m,7H),3.12–2.96(m,10H),1.81(s,9H),1.73(s,12H),1.57–1.35(m,31H),1.29–1.24(m,12H).

[0359] Preparation of Example Compound A43

[0360]

[0361] The synthesis of compound A43 was prepared by referring to compound A42. MS: 879.4 (1 / 2 [M-DMTr] + H) + . 1H NMR (400MHz, DMSO-d6) δ7.72(t,J=5.7Hz,3H),7.62(d,J=9.0Hz,3H),7.50(t,J=5.9Hz,4H),7.36–7.28(m,4H),7.21(td,J=9.0,4.1Hz,5H),6. 92–6.87(m,4H),4.95(dd,J=32.7,4.2Hz,1H),4.64–4.53(m,6H),4.48( d,J=4.3Hz,3H),4.42(d,J=4.7Hz,1H),4.34–4.09(m,4H),3.79–3.64(m ,16H),3.55(ddh,J=16.3,10.9,5.2Hz,7H),3.47–3.41(m,3H),3.40–3 .36(m,2H),3.33–3.29(m,3H),3.27–3.16(m,1H),3.03(q,J=6.3Hz,16H ), 2.22(t,J=7.4Hz,1H),2.05(t,J=7.4Hz,7H),1.86(d,J=8.3Hz,1H),1.81(s,9H),1.72(s,11H),1.53–1.35(m,29H),1.22(d,J=12.9Hz,19H).

[0362] Example: Aminogalacin compound III-LA16-CPG attached to a solid support

[0363] The aminogalactose compound connected to the solid phase carrier disclosed in the present invention is synthesized by methods well known to those skilled in the art or by methods described in detail in the prior art. For example, WO2015006740A2 describes in detail the preparation method of the aminogalactose compound connected to the solid phase carrier.

[0364]

[0365] Compound III-LA16-1; MS: 1076.7 (1 / 2 [M-DMTr] + H) + . 1H NMR(400MHz, DMSO-d6)δ7.82(d,J=9.2Hz,2H),7.79–7.69(m,3H),7.55(s,3H),7.37–7.25(m,4H),7.20(ddt,J=9.0,5.9,2.3Hz,5H),6.87( ddd,J=8.8,5.4,2.4Hz,4H),5.21(d,J=3.4Hz,3H),4.97(dd,J=11.2,3.4Hz,2H),4.49(d,J=8.4Hz,2H),4.05–3.97(m,9H),3.87(dt,J=11. 3,8.9Hz,4H),3.73(s,12H),3.55(d,J=11.0Hz,4H),3.40(s,5H),3.09–2.91(m,17H),2.48–2.38(m,3H),2.21(s,2H),2.10(s,9H),2.04(t ,J=7.2Hz,7H),1.99(s,9H),1.89(s,9H),1.77(s,10H),1.72(s,11H),1.48(dt,J=15.3,7.1Hz,22H),1.21(s,12H),0.96(t,J=7.1Hz,9H).

[0366] Example: Aminoglucosyl compound III-LA20-CPG attached to a solid support

[0367] The synthesis of the aminogalactose compound III-LA20-CPG linked to the solid phase support was based on the synthesis of the aminogalactose compound III-LA16-CPG linked to the solid phase support.

[0368]

[0369] Compound III-LA20-1; MS: 1093.8 (1 / 2 [M-DMTr] + H) + . 1H NMR (400MHz, DMSO-d6) δ7.97(s,1H),7.84(d,J=9.2Hz,3H),7.72(d,J=5.7Hz,2H),7.32(d,J=11.9Hz,4H),7.22(t,J=8.0Hz,5H),6.88(d,J= 7.3Hz,3H),5.37(s,0H),5.23(d,J=3.3Hz,3H),4.99(dd,J=11.4,3.4Hz,2H),4.51(d,J=8.4Hz,2H),4.22(s,1H),4.05(d,J=7.9Hz,10H),3.8 9(q,J=9.6Hz,5H),3.75(s,24H),3.42(d,J=7.9Hz,7H),3.34(s,4H),3.19(s,4H),3.02(d,J=7.3Hz,8H),2.53(d,J=2.9Hz,4H),2.25(t,J=6 .8Hz,7H),2.12(s,10H),2.09–1.97(m,21H),1.91(s,9H),1.79(s,9H),1.57–1.33(m,30H),1.25(d,J=14.4Hz,18H),0.97(t,J=7.0Hz,11H).

[0370] Example: Aminoglucosyl compound III-LA30-CPG attached to a solid support

[0371] The synthesis of the aminogalactose compound III-LA30-CPG linked to the solid phase support was based on the synthesis of the aminogalactose compound III-LA16-CPG linked to the solid phase support.

[0372]

[0373] Compound III-LA30-1; MS: 1065.3 (1 / 2 [M-DMTr] + H) + . 1H NMR(400MHz, DMSO-d6)δ7.82(d,J=9.2Hz,3H),7.71(s,3H),7.38–7.24(m,4H),7.19(ddd,J=9.1,5.9,3.2Hz,5H),6.91–6.80(m ,4H),5.21(d,J=3.4Hz,3H),4.97(dd,J=11.2,3.4Hz,3H),4.49(d,J=8.4Hz,3H),4.02(q,J=4.1Hz,9H),3.87(dt,J=11.3,8.9Hz ,4H),3.80–3.66(m,12H),3.30(s,3H),3.18(s,5H),2.99(d,J=7.1Hz,7H),2.46(dd,J=12.2,5.1Hz,11H),2.21(s,7H),2.10(s, 9H), 2.07–2.00 (m, 8H), 1.99 (s, 10H), 1.89 (s, 9H), 1.77 (s, 9H), 1.40 (d, J = 42.0Hz, 29H), 1.23 (s, 21H), 0.95 (t, J = 7.1Hz, 11H).

[0374] Example Synthesis of siRNA conjugated with aminogalactose clusters

[0375] The siRNA used for the test, siRNA targeting mouse TTR gene mRNA (Nucleic Acids Res. 2020 Dec 2; 48(21): 11827-11844) is as follows: an aminogalactose molecule cluster M is covalently linked to the 3' end of the SS chain,

[0376] SS strand (5'-3'): a·a·caguGuUCUugcucuauaaL

[0377] AS chain (5'-3'):u·U·auaGaGCaagaAcAcuguu·u·u

[0378] Capital letters and lowercase letters represent monomers modified with 2′-deoxy-2′-fluoro (2′-F) and 2′-O-methyl (2′-OMe), respectively. · represents thio, and L represents N-acetylgalactosamine ligand.

[0379] The siRNA conjugates disclosed herein are synthesized by methods well known to those skilled in the art or by methods described in detail in the prior art. For example, WO2015006740A2 describes in detail the preparation methods of various siRNA conjugates.

[0380] Synthesize aminogalactose cluster-conjugated siRNA, and the siRNA used in the experiment targets mouse TTR mRNA.

[0381]

[0382]

[0383] Table X. siRNA numbers and sequences for evaluation of targeting ligand activity

[0384]

[0385] Among them, S-L96 was purchased from Jima Gene; C, G, A, U or c, g, a, u represent the base composition of nucleotides. The uppercase and lowercase letters represent monomers modified with 2′-deoxy-2′-fluoro (2′-F) and 2′-O-methyl (2′-OMe), respectively. · represents thio, and L represents N-acetylgalactosamine ligand.

[0386] After the preparation of the siRNA conjugate disclosed above is completed, it is freeze-dried into solid powder using standard means and stored for later use. When in use, it can be redissolved into a solution of desired concentration using, for example, water for injection, physiological saline, phosphate buffer or phosphate buffer.

[0387] (I) Principle of ASGPR-GalNAc TM affinity detection test based on monkey primary cells: The test compound and GalNAc-Cy5 compete with the ASGPR receptor on the surface of monkey primary cells (PCMH, purchased from Miaoshun Biotechnology Co., Ltd.). The fluorescence intensity of Cy5 detected by flow cytometry can reflect the affinity of the test compound to ASGPR. The weaker the Cy5 fluorescence intensity, the stronger the affinity of the test compound to ASGPR, and vice versa.

[0388] Experimental process: Monkey primary cells were plated in 96-well plates at 40,000 / well and incubated with GalNAc-Cy5 (final concentration 100 nM) and the test compound (final concentration starting from 1 μM, 10-fold dilution for initial screening) in a 37°C, 5% CO2 incubator for 4 hours. The cells were then collected and the fluorescence intensity of Cy5 in the cells was analyzed by flow cytometry.

[0389] Conclusion: The results of affinity detection experiments show that the affinity of most of the compounds currently tested, such as A12, A16, A17, A18, A19, A20, A23, A24, A25, A27, A30, A32, A35, A39, A40, A42, and A43, to ASGPR shows activity comparable to or slightly superior to that of the L96-DeAc molecule.

[0390] (II) Inhibition of mRNA expression by siRNA conjugated with aminogalactose clusters in vivo C57 / BL6 mice aged 6 to 8 weeks were randomly divided into 3 groups, 3 mice in each group, and 1 group without any treatment. The positive control group (S-L96) and drug treatment group (S-LA20, S-LA16, S-LA30) were injected subcutaneously with GalNac-siRNA at a dose of 0.3 mg / kg, 1 mg / kg and 3 mg / kg. Blood was collected from the eye sockets to separate serum before administration (day 0), 3 days after administration (day 3), 7 days after administration (day 7), 14 days after administration (day 14) and 21 days after administration (day 21), and mice were killed on day 21 to obtain liver samples. The TTR levels in mouse serum and liver samples were detected using the Mouse Prealbumin ELISA Kit (cat#ALPCO(41-PALMS-E01)), and the results are as follows:

[0391] Table 1. Detection of TTR protein levels in mouse serum and liver samples

[0392]

[0393]

[0394]

[0395] Conclusion: The in vivo efficacy test data showed that S-LA20, S-LA16, and S-LA30 exhibited excellent mTTR gene expression inhibition efficiency, and the activity levels at 0.3mpk, 1mpk, and 3mpk were comparable to or slightly superior to the control group S-L96.

[0396] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A GalNAc delivery molecule as shown in the following formula (I), or a stereoisomer, enantiomer, or a pharmaceutically acceptable salt thereof: in, for R1 is selected from the group consisting of H, DMTr, phosphate, phosphorothioate, phosphorothioate-siRNA conjugate, or phosphate-siRNA conjugate; Linker has a structure selected from the group consisting of C4-C20 alkylene, wherein n, n3 and m are each independently selected from the following group: 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; n1 and n2 are each independently selected from the following group: 0 or 1; Z is selected from the following group: substituted or unsubstituted C 1-8 Alkylene, substituted or unsubstituted 7-12 membered fused bicyclic ring, substituted or unsubstituted 7-15 membered spirocyclic ring, substituted or unsubstituted 5-15 membered bridged ring, -NH-substituted or unsubstituted 7-12 membered fused bicyclic ring, -NH-substituted or unsubstituted 7-15 membered spirocyclic ring, -NH-substituted or unsubstituted 5-15 membered bridged ring, -NH-substituted or unsubstituted 7-12 membered fused bicyclic ring-NH-, -NH-substituted or unsubstituted 7-15 membered spirocyclic ring-NH-, -NH-substituted or unsubstituted 5-15 membered bridged ring-NH-; wherein the ring atoms of the fused bicyclic ring, spirocyclic ring and bridged ring may be optionally carbon atoms or heteroatoms; The substitution refers to the substitution of one or more hydrogen atoms on the group by a substituent selected from the following group: 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, 3- to 12-membered heterocyclic group, C 6-10 Aryl, 5- to 10-membered heteroaryl, halogen, hydroxyl, carboxyl (-COOH), C 1-8 Aldehyde, C 2-10 Acyl, C 2-10 Ester group, amino group, C 1-8 Alkoxy, C 1-10 Sulfonyl; or two substituents located on adjacent ring atoms may together with the connected ring atoms form a group selected from the following group: a 5-7 membered carbon ring or heterocyclic ring, a benzene ring, or a 5-7 membered heteroaromatic ring; Q is selected from the group consisting of a chemical bond, -NH-C(O)-, -C(O)-; r is 0, 1, 2, 3, 4, 5 or 6; R2 is a GalNAc target head portion and has a structure selected from the group consisting of: Wherein, L1 and L2 are independently structures as shown below: L3, L4, L5, and L6 are each independently a structure shown in the following formula: Among them, R 11 Selected from the following group: H or C 1-4 alkyl; Each u, v and w is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11; Glu is an unmodified or modified 5-6-membered sugar group; X1 is selected from the following group: substituted or unsubstituted C 4-10 a saturated or partially unsaturated carbocyclic group, a substituted or unsubstituted 4-10 membered saturated or partially unsaturated heterocyclic group (preferably a nitrogen-containing heterocyclic group), a substituted or unsubstituted C 6-10 Phenyl, or substituted or unsubstituted 5-10 membered heteroaryl; X2 is selected from the following groups: -NH-(C1-C6 alkyl)-NH-, -NH-(C3-C8 cycloalkyl)-NH-, -NH-(4-10 membered heterocyclyl)-NH-, substituted or unsubstituted C 4-10 a saturated or partially unsaturated carbocyclic group, a substituted or unsubstituted 4-14 membered saturated or partially unsaturated heterocyclic group (preferably a nitrogen-containing heterocyclic group), a substituted or unsubstituted C 6-10 Phenyl, or substituted or unsubstituted 5-10 membered heteroaryl; Wherein, the carbocyclic group and heterocyclic group may be a single ring, a condensed ring, a bridged ring or a spiro ring; and R3, R4 and R5 are each independently a substituted or unsubstituted sugar group, preferably a group formed by a substituted or unsubstituted N-acetylgalactosamine molecule; Selected from the group consisting of: substituted or unsubstituted C 4-15 a saturated or partially unsaturated carbon ring group, a substituted or unsubstituted 4-12 membered saturated or partially unsaturated nitrogen-containing heterocyclic group, a substituted or unsubstituted C 6-18 Aryl, substituted or unsubstituted 5-18 membered heteroaryl; The substitution refers to the substitution of one or more hydrogen atoms on the group by a substituent selected from the following group: 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, 3- to 12-membered heterocyclic group, C 3-8 Aryl, 5- to 7-membered heteroaryl, halogen, hydroxyl, carboxyl (-COOH), C 1-8 Aldehyde, C 2-10 Acyl, C 2-10 Ester group, amino group, C 1-8 Alkoxy, C 1-10 or two substituents located on adjacent ring atoms may together with the connected ring atoms form a group selected from the following group: a 5-7 membered carbon ring or heterocyclic ring, a benzene ring, or a 5-7 membered heteroaromatic ring.

2. The GalNAc delivery molecule according to claim 1, or a stereoisomer, an enantiomer, or a pharmaceutically acceptable salt thereof, wherein: The linker is a C4-C20 alkylene group.

3. The GalNAc delivery molecule according to claim 1, or a stereoisomer, an enantiomer, or a pharmaceutically acceptable salt thereof, wherein: The L1, L2 and L3 are each independently a structure shown in the following formula: The L3, L4, L5 and L6 are each independently a structure shown in the following formula:

4. The GalNAc delivery molecule according to claim 1, or a stereoisomer, an enantiomer, or a pharmaceutically acceptable salt thereof, wherein: The sugar group has a structure shown in the following formula III: Among them, the R 15 Selected from the group consisting of -NH(C2-C6 acyl), -NH(halogenated C2-C6 acyl), -NH(C2-C6 sulfonyl), -NH(halogenated C2-C6 sulfonyl); R 12 , R 13 and R 14 Each is independently selected from the following group: H or C1-C6 acyl.

5. The GalNAc delivery molecule according to claim 4, or a stereoisomer, an enantiomer, or a pharmaceutically acceptable salt thereof, wherein: The R 15 Selected from the group consisting of: -NH(C2-C4 acyl); R 12 , R 13 and R 14 Each is independently selected from the following group: H or C1-C4 acyl.

6. The GalNAc delivery molecule according to claim 4, or a stereoisomer, an enantiomer, or a pharmaceutically acceptable salt thereof, wherein: The R 15 is -NHAc; R 12 , R 13 and R 14 Each is independently selected from the group consisting of H or Ac.

7. The GalNAc delivery molecule according to claim 1, or a stereoisomer, an enantiomer, or a pharmaceutically acceptable salt thereof, wherein: The is a substituted or unsubstituted structure selected from the following group: a substituted or unsubstituted 9-18 membered nitrogen heterocycle, a substituted or unsubstituted C 4-12 Monocycloalkyl, substituted or unsubstituted C6-15 bridged cycloalkyl.

8. The GalNAc delivery molecule according to claim 1, or a stereoisomer, an enantiomer, or a pharmaceutically acceptable salt thereof, wherein: The is a substituted or unsubstituted structure selected from the following group:

9. The GalNAc delivery molecule according to any one of claims 1 to 8, or a stereoisomer, enantiomer, or a pharmaceutically acceptable salt thereof, characterized in that: The GalNAc delivery molecule has a structure selected from the group consisting of:

10. The GalNAc delivery molecule according to any one of claims 1 to 8, or a stereoisomer, enantiomer, or a pharmaceutically acceptable salt thereof, characterized in that: The GalNAc delivery molecule has a structure selected from the group consisting of:

11. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: one or more of the compound of formula I according to any one of claims 1 to 10, its pharmaceutically acceptable salt, racemate, R-isomer, S-isomer or mixture thereof, and one or more pharmaceutically acceptable carriers, excipients, adjuvants, auxiliary materials and / or diluents.

12. The use of a compound of formula I as claimed in any one of claims 1 to 10, its pharmaceutically acceptable salt, racemate, R-isomer, S-isomer or mixture thereof, characterized in that: Used for preparing a pharmaceutical composition for treating or preventing tumors or infections caused by viruses.

13. Use of a compound of formula (I) according to any one of claims 1 to 10 for preparing a medicament for treating and / or preventing a disease or condition selected from the group consisting of a disease or condition caused by hepatitis B virus, a disease or condition caused by abnormal expression of ANGPTL3 gene, a disease or condition caused by abnormal expression of TTR gene, a disease or condition caused by abnormal expression of hepatocyte genes, or other liver diseases or conditions.

14. A GalNAc delivery molecule linker as shown in formula (Ia), or a stereoisomer, enantiomer, or a pharmaceutically acceptable salt thereof: The part shown in formula (Ia) has a structure selected from the group consisting of in, It is the site connected to R1.

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