Drug-loaded macromolecules and their preparation methods

Abstract

A class of drug-loaded macromolecules and their preparation methods are disclosed. Specifically, the macromolecules are dendritic polymers loaded with drugs and pharmacokinetic modifiers, particularly involving the linking of drugs to dendritic polymers via specific linkers. These macromolecules can be used to regulate drug release rates, specifically through the selection of linkers.

Description

Technical Field

[0001] This disclosure pertains to the pharmaceutical field and relates to dendritic polymers loaded with drugs and pharmacokinetic modifiers, particularly to the linking of drugs to dendritic polymers via specific linkers. Background Technology

[0002] While significant progress has been made in drug research and development, many drugs still fail in clinical trials due to physical properties (such as solubility) that make them difficult to formulate into suitable dosage forms, or due to toxic effects and poor therapeutic indexes caused by high drug concentrations after administration. Other drawbacks include poor absorption, low bioavailability, poor in vivo stability, poor targeting leading to systemic side effects, and the inability to control biodistribution, metabolism, and renal or hepatic clearance after administration. With continuous advancements in drug research, new research areas and promising technological approaches have emerged to advance drug development, such as formulating drug agents in liposomes, micelles, or polymeric micelle formulations, and covalently attaching drug agents to hydrophilic polymer backbones. While these strategies can solubilize active pharmaceutical ingredients and, in some cases, improve bioavailability and targeting, the release of the active pharmaceutical ingredient remains challenging. In some cases, the carrier rapidly degrades, releasing the active pharmaceutical ingredient, before the drug molecule reaches the target organ. In many cases, the release rate of the active pharmaceutical ingredient from the carrier is variable, thus preventing the drug from reaching an effective therapeutic dose in vivo or at the target organ.

[0003] In recent years, dendritic polymers have shown significant progress in biotechnology and pharmaceutical applications (Xiangyang Shi et al., Sci China Mater, 2018, 61(11), 1387–1403). Dendritic polymers are a specific type of polymer with a densely branched structure. They are macromolecules that grow outwards from a core molecule through repeated branching, resulting in a tree-like structure. The core branches to a certain length and then divides into two branches, repeating this process until the structure becomes dense enough to resemble a spherical clump (V Gajbhiye et al., Journal of Pharmacyanid Pharmacology, 2009, 61, 989–1003). They are characterized by a higher concentration of functional groups per unit molecular volume than ordinary polymers. In particular, the unique properties of dendritic polymers, such as their high degree of branching, multivalent nature, spherical structure, and well-defined molecular weight, offer promise for novel scaffolds in drug delivery. Over the past decade, research on the design and synthesis of biocompatible dendritic polymers and their applications in many areas of bioscience, including drug delivery, has received increasing attention.

[0004] Australian company Starpahrma utilizes its developed dendritic polylysine for loading and transporting anticancer drugs, enhancing their pharmacological properties and ensuring they are delivered to the appropriate site in the body at the right time. This method is known as "drug delivery." The trademark was launched. Among them... as well as This is what the company currently utilizes The technology focuses on developing three anticancer drugs that are in the clinical research stage, showing great development prospects.

[0005] CN103796684A discloses a macromolecule that links a drug to a dendritic polymer via a diacid linker, particularly via a diacid linker containing saturated branched or straight-chain C1-C10 chains interrupted by oxygen, nitrogen, or sulfur atoms. Summary of the Invention

[0006] This disclosure provides a macromolecule comprising:

[0007] i) A dendritic polymer D having surface amino groups, wherein at least two distinct end groups are covalently linked to the surface amino groups of the dendritic polymer:

[0008] ii) The first terminal group is a pharmaceutically active agent containing a hydroxyl, amino, or thiol group or residue A thereof;

[0009] iii) The second end group, which is a pharmacokinetic modifier;

[0010] The first end group is covalently linked to the surface amino group of the dendritic polymer via a linker -X1-L-X2-, where X1 is the linker to the pharmaceutical active agent or its residue A, X2 is the linker to the dendritic polymer D, and both X1 and X2 are -C(O)-, and L is C 1-10 Straight-chain or branched alkylene groups, wherein the C 1-10 Straight-chain or branched alkylene groups are selected from deuterium, hydroxyl, C 3-7 cycloalkyl, C 3-7 Cycloalkylene, C 1-6 The alkoxy group is substituted with one or more of the following: alkoxy, haloalkyl, haloalkoxy, halogen, nitro, cyano, acyl, mercapto, thioether, sulfinyl, sulfonyl, -NR1R2, aryl, heteroaryl, and heterocyclic.

[0011] R1 and R2 are each independently selected from hydrogen, hydroxyl, and C. 1-6 Alkyl, cycloalkyl, C 1-6 Alkyl group.

[0012] In some implementations, L is C 1-10Straight-chain or branched alkylene, wherein C 1-10 The straight-chain or branched alkylene groups are substituted with one or more substituents selected from deuterium, halogens, -OR1, -SR1, -NR1R2 and -C(O)R3;

[0013] R1 and R2 are each independently selected from hydrogen, hydroxyl, and C. 1-6 Alkyl, C 3-7 cycloalkyl, C 1-6 Alkoxy and C(O)R4, the C 1-6 Alkyl, C 3-7 cycloalkyl and C 1-6 Alkoxy groups are optionally selected from hydroxyl, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, nitro, cyano, amino and C 1-6 One or more substituents in the alkylamino group are substituted;

[0014] R3 and R4 are each independently selected from hydrogen and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 1-6 Alkyl group.

[0015] In some implementations, R1 is not hydrogen.

[0016] In some implementations, the C 1-10 The straight-chain or branched alkylene group is substituted with one or more substituents selected from -OR1, -SR1, -NR1R2, wherein R1 and R2 are each independently selected from hydrogen, C 1-6 Alkyl, C 3-7 cycloalkyl, C 1-6 Alkoxy and C(O)R4, the C 1-6 Alkyl, C 3-7 cycloalkyl and C 1-6 Alkoxy groups are optionally selected from hydroxyl, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, amino and C 1-6 One or more substituents in the alkylamino group are substituted, and R4 is selected from hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 1-6 Alkyl group.

[0017] In some implementations, the C 1-10The straight-chain or branched alkylene group is substituted with one or more -NR1R2, wherein R1 and R2 are each independently selected from hydrogen, C 1-6 Alkyl, C 3-7 cycloalkyl, C 1-6 Alkoxy and C(O)R4, the C 1-6 Alkyl, C 3-7 cycloalkyl and C 1-6 Alkoxy groups are optionally selected from hydroxyl, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, amino and C 1-6 One or more substituents in the alkylamino group are substituted, and R4 is selected from hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 1-6 Alkyl group.

[0018] In some implementations, the C 1-10 The straight-chain or branched alkylene group is substituted with one or more -NR1R2, wherein R1 and R2 are each independently selected from hydrogen, C 1-6 Alkyl groups and C(O)R4, where R4 is selected from hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 1-6 Alkoxy group; preferably R1 and R2 are each independently selected from hydrogen and C. 1-6 Alkyl group, and R1 and R2 are not both hydrogen.

[0019] In some implementations, the C 1-10 Straight-chain or branched alkylene groups are C 1-6 Straight-chain or branched alkylene groups.

[0020] In some embodiments, in the linker -X1-L-X2-, X1 is -C(O)- and is linked to the pharmaceutical active agent or its residue A; X2 is -C(O)- and is linked to the surface amino group of the dendritic polymer D to form an amide bond.

[0021] In some embodiments, the pharmaceutically active agent or its residue A contains a hydroxyl group and forms an ester bond with X1.

[0022] In some specific embodiments, the macromolecular structure is selected from the following:

[0023]

[0024]

[0025] In some embodiments, selecting the linker of this disclosure can provide the desired drug release rate, such as rapid or slow release.

[0026] In some embodiments, the release rate of the macromolecular pharmaceutical active agent is faster than that of the macromolecular-independent agent, possibly at least twice as fast. In some embodiments, the release rate of the macromolecular pharmaceutical active agent is slower than that of the macromolecular-independent agent, possibly two, three, four, five, six, seven, eight, nine, ten, or more than ten, fifteen, twenty, or thirty times slower. Macromolecules with low release rates are suitable for formulation into drugs that release slowly over long periods, such as 1 week to 3 months, 1 month to 6 months, or more than 6 months. Rapid release is preferably within 0-8 hours, especially within 0-4 hours, particularly within 0-2 hours, and even more particularly within 5-60 minutes, releasing more than 50% of the pharmaceutical active agent. Moderate release is preferably within 1-72 hours, especially within 2-48 hours, releasing more than 50% of the pharmaceutical active agent. The release rate of the pharmaceutical active agent can be controlled by selecting a suitable linker, and the release rate also depends on the characteristics of the pharmaceutical active agent. In some embodiments, the pharmaceutical active agent is linked to the dendritic polymer via the same linker. In other embodiments, the pharmaceutical active agent is linked to the dendritic polymer via two or more linkers, allowing the pharmaceutical active agent to be released from the macromolecule at different release rates.

[0027] In some embodiments, the first end group and the second end group are present in a ratio of 1:2 to 2:1, particularly 1:2, 1:1, or 2:1. In some embodiments, the macromolecule includes a third end group as a blocking group, a pharmaceutical group, or a targeting group. The blocking group may be an acyl group. In some embodiments, the ratio of the first, second, and third end groups is 1:1:1 to 1:2:2, particularly 1:2:1. In some embodiments, at least 50% of the end groups contain either the first or second end group. In specific embodiments, the pharmaceutically active agent is bound to more than 14%, 25%, 27%, 30%, 39%, 44%, or 48% of the surface amino groups. In some embodiments, the pharmacokinetic modifier is bound to more than 15%, 25%, 30%, 33%, or 46% of the surface amino groups.

[0028] The pharmaceutically active agents described in this disclosure may be selected from: anesthetics, antacids, antibodies, anti-infectives, biological products, cardiovascular drugs, contrast agents, diuretics, hematologic agents, immunosuppressants, hormones and analogues, nutritional products, ophthalmic drugs, pain management agents, respiratory drugs, adjuvants, anabolic agents, anti-arthritis drugs, anticonvulsants, antihistamines, anti-inflammatory drugs, anti-ulcer drugs, behavioral modification drugs, oncology drugs, central nervous system drugs, contraceptives, diabetes treatment drugs, fertility drugs, growth promoters, hemostatic agents, immunostimulants, muscle relaxants, obesity treatment agents, osteoporosis drugs, peptides, sedatives and tranquilizers, urethral acidifiers, or vitamins.

[0029] In some implementation schemes, the active pharmaceutical ingredient is an oncology drug, a steroid, an opioid analgesic, a respiratory drug, a central nervous system (CNS) drug, a drug for hypercholesterolemia, an antihypertensive drug, an antibacterial agent, an immunosuppressive drug, an antibiotic, a luteinizing hormone-releasing hormone (LHRH) agonist, an LHRH antagonist, an antiviral drug, an antiretroviral drug, an estrogen receptor modulator, a somatostatin analog, an anti-inflammatory drug, a vitamin D2 analog, a synthetic thyroxine, an antihistamine, an antifungal agent, or a nonsteroidal anti-inflammatory drug (NSAID), preferably an oncology drug.

[0030] In some embodiments, the oncology drugs include taxanes (such as paclitaxel, cabazitaxel, and docetaxel), camptothecin and its analogues (such as irinotecan and topotecan), nucleoside analogues (such as gemcitabine, cladribine, fludarabine, capecitabine, decitabine, azacitidine, clofarabine, and nelabine), and kinase inhibitors (such as dasatinib (sprycel), tamsirolimus, AZD6244, AZD1152, PI-103, R-roscovitine, olomocin, and purvalanol). A) and epothilone B analogues (such as ixaprone), anthrocyclines (such as amrubicin, doxorubicin, epirubicin and pentorubicin), saginoid derivatives (such as trabectecin and lurbinectedin), proteasome inhibitors (such as bortezomib) and other topoisomerase inhibitors, intercalating agents and alkylating agents, microtubule inhibitors (such as eribulin and vinflunine), or structural modifications of these drug molecules.

[0031] In some embodiments, the pharmaceutically active agent is selected from taxanes, camptothecin derivatives, nucleoside analogs, anthracycline drugs, sucralose derivatives, proteasome inhibitors, microtubule inhibitors, BCL-2 inhibitors, and BCL-X. L Inhibitors, selective nuclear export inhibitors, antimetabolites, tyrosine kinase inhibitors, PLK1 inhibitors, CDK4 / 6 inhibitors, BTK inhibitors, nonsteroidal hormone receptor antagonists, and steroids, preferably taxanes, camptothecin derivatives, BCL-2 inhibitors, or BCL-X. L Inhibitors.

[0032] In some embodiments, the pharmaceutically active agent is selected from docetaxel, irinotecan, gemcitabine, capecitabine, decitabine, azacitidine, doxorubicin, epirubicin, trabectedin, rupettedin, bortezomib, eribulin, selinexol, venetoclax, tercitabine, pemetrexed, cabazitaxel, cabozantinib, onvansertib, compounds 2 and 3 below, or structural modifications of these drug molecules.

[0033]

[0034] In some embodiments, the steroids include synthetic steroids (such as testosterone, dihydrotestosterone, and ethinylestradiol) and corticosteroids (such as cortisone, prednisilone, budesonide, triamcinolone, fluticasone, mometasone, ansine, flucinolone, fluocinanide, desonide, cloxacillin, prednisolone, flucloxacillin, dexamethasone, betamethasone, and fluprednidine).

[0035] In some embodiments, the opioid analgesics include morphine, oxymorphone, naloxone, codeine, oxycodone, methylnaltrexone, hydromorphone, buprenorphine, and etorphine. In some embodiments, the respiratory medications include bronchodilators, inhaled steroids, and decongestants, particularly salbutamol, ipratropium bromide, montelukast, and formoterol. In some embodiments, the CNS medications include antipsychotics (such as quetiapine) and antidepressants (such as venlafaxine). In some embodiments, the hypercholesterolemia medications include ezetimibe and statins such as simvastatin, lovastatin, atorvastatin, fluvastatin, pitavastatin, pravastatin, and rosuvastatin. In some embodiments, the antihypertensive medications include losartan, olmesartan, metronidazole, metoprolol, travoprost, and bosentan. In some embodiments, the immunosuppressive drugs include glucocorticoids, cell inhibitors, antibody fragments, antiimmunophiles, interferons, TNF-binding proteins, and particularly calcineurin inhibitors such as tacrolimus, mycophenolic acids and their derivatives such as mycophenolate mofetil and cyclosporine. In some embodiments, the antimicrobial agents include antibiotics such as amoxicillin, meropenem, and clavulanic acid. In some embodiments, the LHRH agonists include goserelin acetate, dilorelin, and leuprorelin. In some embodiments, the LHRH antagonists include cetrorex, ganirex, abalix, and degarelix. In some embodiments, the antiviral drugs include nucleoside analogues such as lamivudine, zidovudine, abacavir, and entecavir, and antiretroviral drugs include protease inhibitors such as atazanavir, lapinavir, and ritonavir. In some embodiments, the estrogen receptor modulators include raloxifene and fulvestrant. In some embodiments, the somatostatin analogue includes octreotide. In some embodiments, the anti-inflammatory drug includes mesalazine and suitable NSAIDs include paracetamol (acetaminophen). In some embodiments, the vitamin D2 analogue includes paricalcitol. In some embodiments, the synthetic thyroxine includes levothyroxine. In some embodiments, the antihistamine includes fexofenadine. In some embodiments, the antifungal agent includes azoles such as voriconazole.

[0036] In some embodiments, the pharmaceutically active agent is eribulin. In some embodiments, the pharmaceutically active agent is docetaxel. In some embodiments, the pharmaceutically active agent is trabectedin. In some embodiments, the pharmaceutically active agent is lurbinectedin.

[0037] In some embodiments, the pharmaceutically active agent is slightly soluble or insoluble in aqueous solution.

[0038] The second end group is a pharmacokinetic modifier that can modify or modulate the pharmacokinetic characteristics of the pharmaceutical active agent or macromolecule, including absorption, distribution, metabolism, and / or excretion. In a specific embodiment, the pharmacokinetic modifier prolongs the plasma half-life of the pharmaceutical active agent, such that the half-life of the pharmaceutical active agent linked to the macromolecule is longer than that of the pharmaceutical active agent alone or on a non-dendritic polymer carrier. Preferably, the half-life of the macromolecule or composition is at least 2 times longer and more preferably at least 10 times longer than that of the pharmaceutical active agent alone or on a non-dendritic polymer carrier.

[0039] The pharmacokinetic modifier may be selected from polyethylene glycol, polyethyloxazoline, polyvinylpyrrolidone, polypropylene glycol, folate, or folate derivatives relating to ligands of cell surface receptors. In some embodiments, the pharmacokinetic modifier is polyethylene glycol. In some embodiments, the polyethylene glycol has a molecular weight in the range of 220 to 5500 Da, for example, it may be 220-2500 Da, 570-2500 Da, 220-1100 Da, 570-1100 Da, 1000-5500 Da, 1000-2500 Da, or 1000-2300 Da. In some embodiments, the pharmacokinetic modifier forms an amide bond with an amino group on the surface of the dendritic polymer.

[0040] Targeting groups are agents that bind to selective biological target cells, organs, or tissues, thereby facilitating the direction of macromolecules toward specific targets in vivo and their accumulation in those target cells, organs, or tissues. Additionally, targeting groups can provide mechanisms for macromolecules to actively enter cells or tissues via receptor-mediated endocytosis. Specific examples include lectins and antibodies, as well as other ligands (including small molecules) targeting cell surface receptors. This interaction can occur through any type of bond or association (including covalent, ionic, hydrogen, and van der Waals forces). Suitable targeting groups include those that bind to cell surface receptors, such as folic acid receptors, adrenergic receptors, growth hormone, luteinizing hormone receptors, estrogen receptors, epidermal growth factor receptors, fibroblast growth factor receptors (e.g., FGFR2), IL-2 receptors, CFTR, and vascular endothelial growth factor (VEGF) receptors.

[0041] In some embodiments, the targeting group is luteinizing hormone-releasing hormone (LHRH) or a derivative thereof bound to the LHRH receptor. In some embodiments, the targeting group is LYP-1, a peptide that targets the lymphatic system of tumors rather than the lymphatic system of normal tissues. In some embodiments, the targeting group may be an RGD peptide. RGD peptides are peptides containing the sequence -Arg-Gly-Asp-, which is a major integrin recognition site in extracellular matrix proteins. In some embodiments, the targeting group may be folic acid. Estrogen can also be used on target cells expressing estrogen receptors.

[0042] In some embodiments, the targeting group may be directly or preferably bound to the dendritic polymer core via a linking group. The linking group may be any divalent group capable of binding to both the functional groups on the core and the functional groups on the targeting group.

[0043] The macromolecules disclosed herein comprise dendritic polymers, wherein the outermost generation of the structural units has surface amino groups. The properties of the dendritic polymer in the macromolecule are not particularly important, provided that it has surface amino groups. For example, the dendritic polymer can be a polylysine, a polylysine analog, a polyamide (PAMAM), a polyethyleneimine (PEI), or a polyether hydroxylamine (PEHAM) dendritic polymer. In some embodiments, the dendritic polymer is polylysine or a polylysine analog. The polylysine or polylysine analog comprises a core and 2-7 generation lysine or lysine analogs, for example, comprising 2nd, 3rd, 4th, 5th, 6th, or 7th generation lysine or lysine analogs.

[0044] In some embodiments, the lysine has the structure shown in Figure 1:

[0045] In some embodiments, the lysine analogue has the structure shown in Figure 2:

[0046] In some embodiments, the lysine analogue has the structure shown in Figure 3: Where a is 1 or 2; b and c are the same or different and are integers from 1 to 4.

[0047] In some embodiments, the lysine analogue has the structure shown in Figure 4: Where a is an integer from 0 to 2; b and c are the same or different and are integers from 2 to 6.

[0048] In some embodiments, the lysine analogue has the structure shown in Figure 5: Where a is an integer from 0 to 5; b and c are the same or different and are integers from 1 to 5.

[0049] In some embodiments, the lysine analogue has the structure shown in Figure 6: Where a is an integer from 0 to 5; b and c are the same or different and are integers from 0 to 5.

[0050] In some embodiments, the lysine analogue has the structure shown in Figure 7: Where a is an integer from 0 to 5; b and c are the same or different and are integers from 1 to 5.

[0051] In some embodiments, the lysine analogue has the structure shown in Figure 8: Where a is an integer from 0 to 5; b, c, and d are the same or different and are integers from 1 to 5.

[0052] In some embodiments, the lysine analogue has the structure shown in Figure 9: Where a is an integer from 0 to 5; b and c are the same or different and are integers from 1 to 5.

[0053] The dendritic polymers described in this disclosure, particularly polylysine or polylysine analogs, may have a core selected from diphenylmethylamine (BHA), diphenylmethylamine of lysine (BHALys), or lysine analogs, or:

[0054] Where a is 1 to 9, preferably an integer from 1 to 5;

[0055] Where a, b, and c can be the same or different and are integers from 1 to 5, and d is an integer from 0 to 100, preferably an integer from 1 to 30;

[0056] Where a and b can be the same or different, and are integers from 0 to 5;

[0057] Where a and c can be the same or different and are integers from 1 to 6, and b is an integer from 0 to 6;

[0058] Where a and d can be the same or different and are integers from 1 to 6, and b and c can be the same or different and are integers from 0 to 6;

[0059] Where a and b are the same or different and are integers from 1 to 5, especially from 1 to 3, especially 1;

[0060] Where a, b, and c are the same or different, and are integers selected from 1 to 6;

[0061] Where a, b, and c are the same or different, and are integers selected from 0 to 6; Where a, b, and c are the same or different, and are integers selected from 0 to 6;

[0062] Where a, b, and c can be the same or different, and are integers from 0 to 6; d, e, and f can be the same or different, and are integers from 1 to 6.

[0063] Where a, b, and c can be the same or different, and are integers from 1 to 6;

[0064] Where a, b, c, and d can be the same or different, and are integers from 0 to 6;

[0065] Where a, b, c, and d can be the same or different, and are integers from 1 to 6; or

[0066] a, b, c, and d can be the same or different and are integers from 0 to 6; e, f, g, and h can be the same or different and are integers from 1 to 6.

[0067] In some embodiments, the macromolecule comprises:

[0068] i) A dendritic polymer D having surface amino groups, wherein at least two distinct end groups are covalently linked to the surface amino groups of the dendritic polymer:

[0069] ii) The first terminal group is a pharmaceutically active agent containing a hydroxyl, amino, or thiol group or residue A thereof;

[0070] iii) The second end group is polyethylene glycol, a pharmacokinetic modifier;

[0071] The first end group is covalently connected to the surface amino group of the dendritic polymer via a linker, wherein the linker is selected from...

[0072]

[0073] The dendritic polymer D is selected from BHALys[Lys]. 16 BHALys[Lys] 32 Or BHALys[Lys] 64 ,

[0074] The polyethylene glycol has a molecular weight in the range of 1000 to 2500 Da.

[0075] This disclosure also relates to a pharmaceutical composition comprising the macromolecules of this disclosure and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition does not contain solubilizing excipients such as polyethoxylated castor oil or polysorbate. In some embodiments, the pharmaceutical composition is administered via transdermal, oral, or injectable routes.

[0076] The macromolecular formulations of this disclosure include those suitable for oral, rectal, topical, nasal, inhalation, aerosol, ocular, or parenteral (including intraperitoneal, intravenous, subcutaneous, or intramuscular) administration. The compositions are readily available in unit dosage forms and can be prepared by any method well known in the pharmaceutical industry. All methods involve the step of associating the macromolecule with a carrier constituting one or more excipients. Typically, the compositions are prepared by associating the macromolecule with a liquid carrier to form a solution or suspension, or optionally by associating the macromolecule with a formulation component suitable for forming a solid, optionally particulate product, and then, if necessary, shaping the product into the desired delivery form. The solid dosage forms of this disclosure, when in particulate form, will typically comprise a particle size range of about 1 nanometer to about 500 micrometers. Typically, for solid dosage forms intended for intravenous administration, the particle diameter range will typically be about 1 nm to about 10 micrometers. The composition may contain the macromolecules disclosed herein, said macromolecules being nanoparticles having a particle diameter of less than 1000 nm, for example, 5 to 1000 nm, particularly 5 to 500 nm, especially 5 to 400 nm (e.g., 5 to 50 nm and particularly 5 to 20 nm). In certain embodiments, the composition contains macromolecules having an average size of 5 to 20 nm. In some embodiments, the macromolecules are polydisperse in the composition, with a PDI between 1.01 and 1.8, particularly between 1.01 and 1.5, and especially between 1.01 and 1.2. In certain embodiments, the macromolecules are monodisperse in the composition. Particularly preferred are sterile, lyophilized compositions reconstituted in an aqueous medium prior to injection.

[0077] In some embodiments, the composition contains macromolecules having an average size of 5 to 20 nm. In some embodiments, the particle size D of the macromolecules is... 90 Or D 50 Less than 1000 nm, for example, 5 to 1000 nm, particularly 5 to 500 nm, especially 5 to 400 nm (e.g., 5 to 50 nm, particularly 5 to 20 nm). In a particular embodiment, the composition contains having D 50 Large molecules ranging from 5 to 20 nm in size.

[0078] The macromolecules disclosed herein can also be used to provide controlled-release and / or sustained-release formulations of pharmaceutically active agents. In sustained-release formulations, the formulation components are selected to release the macromolecule from the formulation over an extended period (e.g., days, weeks, or months). Such formulations include transdermal patches or implantable devices capable of subcutaneous deposition, or via intravenous, subcutaneous, intramuscular, intradural, or intracranial injection. In controlled-release formulations, a diacid linker is selected to release the majority of its pharmaceutically active agent within a given time window. For example, when the time it takes for the majority of the macromolecule to accumulate in a target organ, tissue, or tumor is known, the linker can be selected after the accumulation time has elapsed to release the majority of its pharmaceutically active agent. This allows for the delivery of a high drug load at the site where its action is required at a given time point. Optionally, the linker is selected to release the pharmaceutically active agent at a therapeutic level over an extended period. In some embodiments, the formulation may have multiple controlled-release properties. For example, a formulation may contain a macromolecule in which the drug is linked by different linkers, allowing for a burst release of the drug followed by a slower release at a lower but constant therapeutic level over an extended period. In some embodiments, the formulation may have sustained-release and controlled-release properties. For example, formulation components may be selected to release the macromolecule over an extended period, and linkers may be selected to deliver a constant, low therapeutic level of the active pharmaceutical ingredient. In some embodiments, the active pharmaceutical ingredient is linked to the same molecule by different linkers. In some embodiments, each drug-linker combination is linked to a different macromolecule in the same formulation.

[0079] In some embodiments, the macromolecules in the pharmaceutical composition are formulated to release more than 50% of the pharmaceutically active agent between 5 minutes and 60 minutes. In some embodiments, the macromolecules in the pharmaceutical composition are formulated to release more than 50% of the pharmaceutically active agent between 2 hours and 48 hours. In some embodiments, the macromolecules in the pharmaceutical composition are formulated to release more than 50% of the pharmaceutically active agent between 5 days and 30 days.

[0080] Another aspect of this disclosure provides a method for treating or inhibiting tumor growth, comprising administering an effective amount of the macromolecular or pharmaceutical composition of this disclosure, wherein the pharmaceutically active agent of the first terminal group is an oncology drug. The tumors described in this disclosure are selected from melanoma, brain tumors, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, breast cancer, ovarian cancer, prostate cancer, skin cancer, neuroblastoma, sarcoma, osteochondroma, osteoma, osteosarcoma, seminoma, testicular tumors, uterine cancer, head and neck tumors, multiple myeloma, malignant lymphoma, polycythemia vera, leukemia, thyroid tumors, ureteral tumors, bladder cancer, gallbladder cancer, bile duct cancer, choriocarcinoma, and pediatric tumors (Ewing familial sarcoma, Wilms' sarcoma, rhabdomyosarcoma, angiosarcoma, embryonal testicular cancer, neuroblastoma, retinoblastoma, hepatoblastoma, nephroblastoma, etc.).

[0081] In another aspect of this disclosure, a method for reducing allergies after treatment with an oncology drug is provided, comprising administering a pharmaceutical composition of this disclosure, wherein the composition is substantially free of solubilizing excipients such as Cremophor EL and polysorbate 80.

[0082] In another aspect of this disclosure, a method for reducing the toxicity of an oncology drug or a formulation thereof is provided, comprising administering a macromolecule of this disclosure, wherein the oncology drug is a first end group of the macromolecule. In some embodiments, the reduced toxicity is hematologic toxicity, neurotoxicity, gastrointestinal toxicity, cardiotoxicity, hepatotoxicity, nephrotoxicity, ototoxicity, or encephalopathy.

[0083] In another aspect of this disclosure, a method for reducing side effects associated with an oncology drug or a formulation of an oncology drug is provided, comprising administering a macromolecule of the present disclosure, wherein the oncology drug is a first end group of the macromolecule. In some embodiments, the reduced side effects are selected from neutropenia, leukopenia, thrombocytopenia, myelotoxicity, myelosuppression, neuropathy, fatigue, nonspecific neurocognitive problems, dizziness, encephalopathy, anemia, taste disturbances, dyspnea, constipation, anorexia, nail disorders, fluid retention, weakness, pain, nausea, vomiting, mucositis, alopecia, skin reactions, myalgia, and allergic reactions.

[0084] In some embodiments, the macromolecular or macromolecular pharmaceutical compositions of this disclosure can reduce or eliminate the need for preoperative medications containing agents such as corticosteroids and antihistamines.

[0085] Methods for preparing dendritic polymers are known in the art. For example, macromolecular dendritic polymers can be prepared by divergent or convergent methods or a combination thereof.

[0086] In the divergent approach, each generation of structural units is sequentially added to the core or the previous generation. Surface units having one or two surface amino groups are protected. If one of the amino groups is protected, the free amino group reacts with one of a linker, a linker-pharmaceutical active agent, or a pharmacokinetic modifier. If both amino groups are protected, and they are protected with different protecting groups, one protecting group can be removed without removing the other. One of the amino protecting groups is removed and reacted with one of a linker, a linker-pharmaceutical active agent, or a pharmacokinetic modifier. Once the initial end group has been attached to the dendrite, the other amino protecting group is removed and another first and second end group is added. These groups are attached to the surface amino groups via amide formation as known in the art.

[0087] In the convergence method, each generation of structural units is built upon the previous generation to form a dendritic motif. Before or after the dendritic motif is attached to the core, the first and second end groups can be attached to the surface amino groups as described above.

[0088] In the hybrid approach, each generation of structural units is added to the core or the previous generation of structural units. However, before the final generation is added to the dendrite, the surface amino groups are functionalized with end groups (e.g., first and second end groups, first and third end groups, or second and third end groups). The functionalized final generation is then added to the subsurface layer of the structural unit, connecting the dendritic motif to the core.

[0089] The pharmaceutical active agent reacts with one of the carboxylic acids of the linker via ester formation as known in the art. For example, an activated carboxylic acid is formed, such as using an acyl chloride or anhydride, and reacts with the hydroxyl group of the pharmaceutical active agent. If the pharmaceutical active agent has more than one hydroxyl group, then the other hydroxyl groups can be protected.

[0090] When the targeting reagent is attached to the core, the functional groups on the core can be protected during dendrite formation and then deprotected, and react with the targeting reagent, the linking group, or the targeting reagent-linking group. Optionally, the core can react with the linking group or the targeting reagent-linking group before dendrite formation.

[0091] Suitable protecting groups, their introduction and removal methods are described in Greene & Wuts, Protecting Groups in Organic Synthesis, 3rd Edition, 1999.

[0092] This disclosure also includes various deuterated forms of macromolecules or pharmaceutically acceptable salts thereof, wherein each available hydrogen atom in the macromolecule can be independently replaced by a deuterium atom. Those skilled in the art will know how to synthesize deuterated forms of the macromolecules or pharmaceutically acceptable salts thereof disclosed herein.

[0093] This disclosure also includes isotopically labeled macromolecules in which one or more atoms are replaced by atoms having a mass or mass number different from the most common atomic masses or mass numbers found in nature. Examples of isotopes that can be used for macromolecules in this disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, iodine, and chlorine, for example... 3 H, 11 C 14 C 18 F, 123 I or 125 I.

[0094] The macromolecules disclosed herein may be in the form of pharmaceutically acceptable salts. However, it should be understood that non-pharmaceutically acceptable salts also fall within the scope of this disclosure, as these may be useful as intermediates in the preparation of pharmaceutically acceptable salts or during storage or transport. Suitable pharmaceutically acceptable salts include, but are not limited to, salts of pharmaceutically acceptable inorganic acids (such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, aminosulfonic acid, and hydrobromic acid) or salts of pharmaceutically acceptable organic acids (such as acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, citric acid, lactic acid, mucoic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, salicylic acid, sulfanilic acid, aspartic acid, glutamic acid, ethylenediaminetetraacetic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid, and valeric acid). Basic salts include, but are not limited to, those that form with pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, ammonium, and alkylammonium.

[0095] Unless otherwise stated, the terms used in the specification and claims shall have the following meanings.

[0096] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0097] "Halogenation" refers to the substitution of one or more atoms selected from fluorine, chlorine, bromine, and iodine.

[0098] "Alkyl" refers to a straight-chain or branched saturated hydrocarbon group having 1 to 10 carbon atoms. Where appropriate, alkyl groups may have a specified number of carbon atoms, for example, C10. 1-4 Alkyl groups, including alkyl groups having 1, 2, 3, or 4 carbon atoms in a straight or branched chain arrangement. Examples of suitable alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 4-methylbutyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 5-methylpentyl, 2-ethylbutyl, 3-ethylbutyl, heptyl, octyl, nonyl, and decyl.

[0099] "Alkylene" refers to a straight-chain or branched divalent alkyl group having 1 to 10 carbon atoms.

[0100] The term "alkenyl" includes branched and straight-chain alkenes having 2 to 12 carbon atoms, or alkenes containing aliphatic hydrocarbon groups, or, if a specific number of carbon atoms is specified, that specific number. For example, "C 2-6"Alkenyl" refers to an alkenyl group having 2, 3, 4, 5, or 6 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl, allyl, 1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methylbut-2-enyl, 3-methylbut-1-enyl, 1-pentenyl, 3-pentenyl, and 4-hexenyl.

[0101] The term "alkynyl" includes branched and straight-chain alkynyl groups having 2 to 12 carbon atoms or alkenes containing aliphatic hydrocarbon groups, or, if a specific number of carbon atoms is specified, that specific number. Examples include ethynyl, propynyl (e.g., 1-propynyl, 2-propynyl), 3-butynyl, pentyynyl, hexynyl, and 1-methylpentan-2-ynyl.

[0102] "Alkenyl" and "ynyl" refer to partially unsaturated branched or straight-chain divalent hydrocarbon groups derived from alken or ynyl groups. In some embodiments, such alkenyl groups are optionally substituted. Non-limiting examples of alkenyl groups include vinylene, propenylene, butenylene, pentenylene, hexenylene, heptenylene, octenylene, nonenylene, decenylene, and similar groups; non-limiting examples of ynylene groups include ethynylene, propynylene, butynylene, pentyynylene, hexynylene, and similar groups.

[0103] "Cycloalkyl" refers to a saturated or unsaturated cyclic hydrocarbon. A cycloalkyl ring may include a specified number of carbon atoms. For example, 3- to 8-membered cycloalkyl groups include 3, 4, 5, 6, 7, or 8 carbon atoms. Suitable examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, 1,4-cyclohexadienyl, cycloheptyl, and cyclooctyl.

[0104] "Cycloalkylene" refers to a divalent cyclic hydrocarbon group derived from a cycloalkyl group. For example... wait.

[0105] The term "alkoxy" refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), where alkyl is defined as described above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexoxy. Alkoxy groups can be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from H atom, D atom, halogen, alkyl, alkoxy, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, and heteroaryl substituents.

[0106] "Aryl" means any stable, monocyclic or bicyclic carbon ring with up to seven atoms in each ring, wherein at least one ring is aromatic. Examples of such aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indenyl, biphenyl, or binaphthyl.

[0107] "Heterocyclic alkyl" or "heterocyclic group" refers to a cyclic hydrocarbon in which one to four carbon atoms have been replaced by heteroatoms independently selected from N, N(R), S, S(O), S(O)2, and O. The heterocycle can be saturated or unsaturated. Examples of suitable heterocyclic groups include tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, pyrrolinyl, pyrazolinyl, pyranyl, piperidinyl, pyrazolinyl, dithiocyclopentadienyl, oxathiocyclopentadienyl, dioxacyclohexyl, dioxacyclohexenyl, morpholinyl, and oxazinyl.

[0108] "Heteroaryl" refers to a stable monocyclic or bicyclic ring with up to 7 atoms in each ring, wherein at least one ring is aromatic and at least one ring contains 1 to 4 heteroatoms selected from O, N and S. The heteroaryl groups within this definition include, but are not limited to, acridinel, carbazolyl, cyclophosphinl, quinoxalinl, quinazolinl, pyrazolyl, indolyl, benzotriazolyl, furanyl, thiophene, phenylthio, 3,4-propylenedioxythiophenyl, benzothiophene, benzofuranyl, benzodioxane, benzodioxane, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, imidazolyl, pyrazinyl, pyridinyl, pyrimidinyl, pyrroleyl, tetrahydroquinoline, thiazolyl, isothiazolyl, 1,2,4-triazolyl, 1,2,3-triazolyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl, 1,3,5-triazinyl, 1,2,4-triazinyl, 1,2,4,5-tetraazinyl, and tetrazolyl.

[0109] A "dendritic polymer" is a molecule containing a core and at least one dendritic unit attached to the core. Each dendritic unit consists of structural units with at least one layer or one generation of branches, resulting in an increasing number of branched structures with branches in each generation of structural units. The maximum number of dendritic units attached to the core is limited by the number of functional groups on the core.

[0110] "Structural unit" refers to a molecule with at least three functional groups, one of which is used to connect to the core or the previous generation of the structural unit and at least two functional groups are used to connect to the next generation of the structural unit or to form the surface of a dendritic polymer.

[0111] "Generation" refers to the number of layers of structural units constituting a dendritic motif or dendritic polymer. For example, a first-generation dendritic polymer will have one layer of structural units connected to the core, such as core-[[structural unit]]u, where u is the number of dendritic motifs connected to the core. A second-generation dendritic polymer has two layers of structural units in each dendritic motif connected to the core. When the structural unit has a branch point, the dendritic polymer can be: core[[structural unit][structural unit]2]u. A third-generation dendritic polymer has three layers of structural units in each dendritic motif connected to the core, such as core-[[structural unit][structural unit]2[structural unit]4]u. A sixth-generation dendritic polymer has six layers of structural units connected to the core, such as core-[[structural unit][structural unit]2[structural unit]4[structural unit]8[structural unit]16[structural unit]32]u, etc. The last generation (outermost generation) of structural units provides the surface functionalization of the dendritic polymer and the number of functionalized groups that can be used to bind end groups. For example, in a dendritic polymer with two dendritic units connected to the core (u=2), if each structural unit has a branch point and there are 6 generations, then the outermost generation has 64 structural units and 128 functional groups that can be used to bind end groups.

[0112] "Slightly soluble" refers to a drug or pharmaceutical active agent that has a solubility of 1 mg / mL to 10 mg / mL in water. Drugs with a solubility of less than 1 mg / mL in water are considered insoluble.

[0113] "Solubilizing excipients" are pharmaceutical additives used to dissolve insoluble or slightly soluble pharmaceutical active agents in aqueous formulations. Examples include surfactants such as polyethoxylated castor oil (including Cremophor EL, Cremophor RH40 and Cremophor RH60), D-α-tocopherol-polyethylene glycol 1000 succinate, polysorbate 20, polysorbate 80, solubilol HS15, sorbitan monooleate, poloxamer 407, and Labrasol.

[0114] "Optional" or "optional" means that the event or environment subsequently described may, but does not have to, occur; the description includes the possibility or possibility that the event or environment may or may not occur. For example, "L is optional C interrupted by one or more oxygen, sulfur, or nitrogen atoms." 1-10 "Linear alkylene" means C 1-10 Straight-chain alkylene groups can be interrupted by oxygen, sulfur, or nitrogen atoms, but are not required to be interrupted. This description includes C. 1-10 The case where straight-chain alkylene groups are interrupted by oxygen, sulfur, or nitrogen atoms and C 1-10 The case where the straight-chain alkylene group is not interrupted by oxygen, sulfur, or nitrogen atoms.

[0115] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort.

[0116] In the chemical structure of the compounds described in this disclosure, the bonds... No configuration was specified, i.e., key It can be or Or simultaneously include and Two configurations. In the chemical structure of the compounds described in this disclosure, the bonds... No configuration is specified, meaning it can be Z configuration, E configuration, or both configurations.

[0117] This disclosure covers the compounds described herein, their pharmaceutically acceptable salts, or any isotopically labeled derivatives of their isomers. Atoms capable of being isotopically labeled include, but are not limited to, hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine. They can be labeled with isotopes. 2 H(D), 3 H, 11 C 13 C 14 C 15 N、 18 F, 31 P, 32 P, 35 S, 36 Cl and 125 I, etc. are used instead. Unless otherwise stated, when a position is specifically designated as deuterium (D), the position shall be understood as having a deuterium abundance of at least 3,000 times greater than the natural abundance of deuterium (which is 0.015%) (i.e., at least 45% deuterium incorporation).

[0118] In this disclosure, the terms "pharmaceutical active agent or its residues", "pharmaceutical active agent", and "pharmaceutical active agent or its residue A" are used interchangeably and all refer to molecules or groups that have pharmaceutical activity. Attached Figure Description

[0119] Figure 1 The pharmacokinetic (PK) curve of the drug in rat plasma in test example 1 is shown.

[0120] Figure 2 The plasma PK curve (0-168 hours) of the beagle dogs in Test Example 2.

[0121] Figure 3The plasma PK curve (0-24 hours) of the beagle dogs in Test Example 2.

[0122] Figure 4 This is a growth curve of tumor volume in each group of mice in the subcutaneous transplantation tumor model of human lung cancer cells A549, as shown in Test Example 3.

[0123] Figure 5 The plasma PK curve of the beagle in test example 4.

[0124] Figure 6 To test the efficacy of the compound in Example 6 against subcutaneous xenografts of human acute lymphoblastic leukemia RS4;11 mice. Detailed Implementation

[0125] The present disclosure is further described and explained below with reference to embodiments, but these embodiments are not intended to limit the scope of the present disclosure.

[0126] Experimental methods not specifying specific conditions in the embodiments of this disclosure are generally performed under conventional conditions or as recommended by the raw material or product manufacturer. Reagents not specifying their source are commercially available, conventional reagents.

[0127] The structure of the compound was determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS). Chemical shifts were expressed as 10⁻⁶. -6 (ppm) is given as the unit.

[0128] MS measurements were performed using a Finnigan LCQAd(ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQadvantage MAX).

[0129] The determination by high performance liquid chromatography (HPLC) was performed using an Agilent 1200DAD high performance liquid chromatograph (Sunfire C18150 × 4.6 mm column) and a Waters 2695-2996 high performance liquid chromatograph (Gimini C18150 × 4.6 mm column).

[0130] Thin-layer chromatography (TLC) uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The silica gel plates used in TLC are 0.15mm to 0.2mm in diameter, while those used for TLC separation and purification are 0.4mm to 0.5mm in diameter.

[0131] Column chromatography typically uses 200-300 mesh silica gel from Yantai Huanghai as the carrier.

[0132] The known starting materials disclosed herein can be synthesized using or in accordance with methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organnics, Aldrich Chemical Company, AccelaChemBio Inc, and Darui Chemicals.

[0133] Unless otherwise specified in the examples, the reactions were carried out under an argon or nitrogen atmosphere.

[0134] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0135] A hydrogen atmosphere refers to a reaction flask connected to a hydrogen balloon with a volume of approximately 1L.

[0136] The pressurized hydrogenation reaction was performed using a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0137] The hydrogenation reaction is usually carried out under vacuum, filled with hydrogen gas, and repeated 3 times.

[0138] The microwave reaction was performed using a CEM Discover-S 908860 microwave reactor.

[0139] Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

[0140] Unless otherwise specified in the examples, the reaction temperature is room temperature.

[0141] Room temperature is the optimal reaction temperature, with a range of 20℃ to 30℃.

[0142] Preparation of PBS buffer solution with pH=6.5 in the example: Take 8.5g of KH2PO4, 8.56g of K2HPO4·3H2O, 5.85g of NaCl and 1.5g of EDTA and put them into a bottle, make up to 2L, sonicate to dissolve completely, and shake well to obtain the solution.

[0143] The eluent systems for column chromatography and the developing solvent systems for thin-layer chromatography used to purify the compounds include: A: dichloromethane and isopropanol system, B: dichloromethane and methanol system, and C: petroleum ether and ethyl acetate system. The volume ratio of the solvents is adjusted according to the polarity of the compounds, and small amounts of triethylamine and acidic or basic reagents can also be added for adjustment.

[0144] Some of the compounds disclosed herein were characterized by Q-TOF LC / MS. The Q-TOF LC / MS was performed using an Agilent 6530 Precision Mass Number Quadrupole-Time-of-Flight Mass Spectrometer and an Agilent 1290-Infinity Ultra-High Performance Liquid Chromatography System (Agilent Poroshell 300SB-C8 5 μm, 2.1 × 75 mm column).

[0145] Referring to the synthesis method of patent CN 110312531A, a type of dendritic polymer was synthesized, in which the first amino terminal group is used for connection with the pharmaceutical active agent, and the second terminal group is used for connection with the pharmacokinetic modifier PEG:

[0146]

[0147] The dendritic polymers illustrated in the following examples include the core of a reference dendritic polymer and structural units in the outermost generation of the dendritic polymer. Generations 1 through subsurface are not depicted. Dendritic polymer BHAL y s[Lys] 32 This represents a fifth-generation dendritic polymer with the following formula: BHALys[Lys]2[Lys]4[Lys]8[Lys] 16 [Lys] 32 64 surface amino groups can be used to bind to end groups.

[0148] Dendritic polymer scaffold BHALys[Lys] 32 [α-NH2·TFA] 32 [ε-PEG 570 ] 32 BHALys[Lys] 32 [α-NH2·TFA] 32 [ε-PEG 1100 ] 32 BHALys[Lys] 32 [α-NH2·TFA] 32 [ε-t-PEG 2300 ] 32 BHALys[Lys] 32 [α-4-HSBA] 32 [ε-PEG 1100 ] 32 BHALys[Lys] 32 [α-GILGVP-NH2.TFA] 32 [ε-PEG 1100 ] 32 And BHALys[Lys] 32 [α-GILGVP-NH2·TFA] 32[ε-t-PEG 2300 ] 32 The preparation can be found in Kaminskas et al., J Control. Release (2011) (doi: 10.1016 / j.jconrel.2011.02.005). Dendritic polymer scaffold 4-azidobenzamide-PEG 12 -NEOEOEN[Su(NPN)2][Lys] 16 [NH2·TFA] 32 The preparation of [the substance] can be found in WO08 / 017122.

[0149] 1-A00, 1-B00, and 1-C00 were synthesized according to patent WO2012167309, with docetaxel as the linked pharmaceutical active agent; 2-A00 was synthesized according to patent WO2018154004A, with compound 2 (synthesized according to patent WO2012017251) as the linked pharmaceutical active agent. Dendritic polymer scaffold BHALys[Lys] 32 [α-NH2·TFA] 32 [ε-PEG 1100 ] 32 In this embodiment, it is referred to as dendritic polymer 1, BHALys[Lys]. 32 [α-NH2·TFA] 32 [ε-PEG 2100 ] 32 In the examples, it is referred to as dendritic polymer 1-PEG2K and synthesized according to the method of WO2018154004A:

[0150]

[0151] General Procedure

[0152] General Procedure A

[0153] Connector and drug installation

[0154] At 0 °C, a coupling agent of EDC or DCC (1.2 equivalents) was added to a magnetically stirred solution of carboxylic acid linkers (0.2–0.5 mmol) in DMF or acetonitrile (1–5 mL). The mixture was stirred for 5 min, and then a solution of a mixture of drug (0.4–1 equivalent) and DMAP (0.4–1 equivalent) in solvent (1 mL) was added dropwise. The mixture was kept at 0 °C for 1 h, and then allowed to warm to ambient temperature. The volatiles were then removed under vacuum, and the residue was purified by preparative HPLC (BEH300 WatersXBridge C18, 5 μM, 30 x 150 mm, 40–80% ACN / water (5–40 min), without buffer) to obtain the desired product.

[0155] General Procedure B

[0156] Connector and drug installation

[0157] Add DIPEA (3 equivalents) to a magnetically stirred solution of drug (0.3–1.0 mmol) and acid anhydride (2 equivalents) in DMF (3–5 mL). Stir the mixture overnight at ambient temperature. Then remove volatiles under vacuum and purify the residue by preparative HPLC (BEH300 Waters XBridge C18, 5 μM, 30 x 150 mm, 40–70% ACN / water (5–40 min), without buffer, RT = 34 min). Concentrate the appropriate fraction under vacuum to obtain the desired product.

[0158] General procedure C

[0159] Dendritic polymers with drug-linker loading

[0160] At room temperature, BHALys[Lys] were magnetically stirred. 32 [α-NH2·TFA] 32 [ε-PEG 1100 ] 32 (0.5–1.0 μmol) and DIPEA (1.2 equimolars / amine) were added to a mixture of DMF along with the linker-drug (1.2 equimolars / amine) and PyBOP (1.2 equimolars / amine). After stirring at room temperature for 1.5 hours, volatiles were removed and the residue was purified by SEC (Sephadex, LH2O, MeOH). Appropriate fractions (as determined by HPLC) were combined and concentrated to obtain the desired product.

[0161] General Procedure D

[0162] Click reaction

[0163] To a magnetically stirred solution of dendritic polymer in 1:1 H₂O / t-BuOH (approximately 0.5 mL) (0.5–1.0 mmol), an alkyne reagent (2 equivalents), sodium ascorbate solution (2 equivalents), and CuSO₄ solution (20 mol%) were added. The solution was heated to 80 °C and monitored by HPLC. Additional amounts of sodium ascorbate and CuSO₄ were added as needed to drive the reaction to completion. After the reaction was deemed complete, the reactants were concentrated under vacuum and then purified.

[0164] The macromolecules disclosed herein can be synthesized via routes selected from the following, where PG is a carboxylic acid protecting group:

[0165]

[0166]

[0167] Example 1: Preparation of Compound 3

[0168]

[0169] first step

[0170] (S)-2-(2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxopropyl-1-en-1-yl)pyrrolidine-1-carboxylic acid tert-butyl ester 3c

[0171] 3a (2.49 g, 7.53 mmol, Shaoyuan) was dissolved in tetrahydrofuran (20 mL), cooled to 0 °C in an ice bath, and N,N-diisopropylaminolithium (661 mg, 6.17 mmol) was added dropwise. After stirring at 0 °C for 1 hour, the reaction system was lowered to -78 °C, and 20 mL of tetrahydrofuran solution of (S)-2-formylpyrrolidine-1-carboxylic acid tert-butyl ester 3b (1.00 g, 5.02 mmol, Yaoshi) was added dropwise. After the addition was complete, the dry ice acetone bath was removed, and the reaction was allowed to proceed to room temperature for 12 hours. The reaction was quenched with saturated ammonium chloride (20 mL), extracted with ethyl acetate (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system B to give the title product 3c (1.90 g, yield: 94.1%).

[0172] MS m / z(ESI): 305.0[M+H-100].

[0173] Step 2

[0174] (2S)-2-(2-amino-3-methoxy-3-oxopropyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3d

[0175] 3c (1.90 g, 4.7 mmol) was dissolved in isopropanol (50 mL), and palladium on carbon (380 mg, 10% purity, dry type) was added. The mixture was purged with hydrogen three times and stirred at room temperature for 8 hours. The reaction solution was filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate (20 mL) and methanol (20 mL). The filtrate was concentrated to obtain the crude product 3d (1.28 g), which was directly proceeded to the next step without further purification.

[0176] Step 3

[0177] (S)-2-((S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-methoxy-3-oxopropyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3e

[0178] (S)-2-((R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methoxy-3-oxopropyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3f

[0179] Crude product 3d (1.28 g, 4.7 mmol) was dissolved in 1,4-dioxane (40 mL), and water (10 mL), sodium bicarbonate (1.17 g, 14.1 mmol), and fluorenemethyloxycarbonyl chloride (1.21 g, 4.7 mmol) were added. The mixture was stirred at room temperature for 3 hours. The solution was quenched with water (30 mL), extracted with ethyl acetate (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography using eluent system A to give the title products 3e (1.03 g, 44.2%, peak 1) and 3f (513 mg, 22.1%, peak 2).

[0180] MS m / z (ESI): 495.2 [M+1].

[0181] Peak 1: UPLC analysis: retention time 2.77 min (column: ACQUITY UPLC BEHC18 50*2.1mm, 1.7um; mobile phase: acetonitrile / water / formic acid = 10 / 90 / 0.1 (v / v / v) to 95 / 5 / 0.1 (v / v / v) gradient elution).

[0182] Peak 2: UPLC analysis: retention time 2.69 min (column: ACQUITY UPLC BEHC18 50*2.1mm, 1.7um; mobile phase: acetonitrile / water / formic acid = 50 / 50 / 0.1 (v / v / v) to 95 / 5 / 0.1 (v / v / v) gradient elution).

[0183] Step 4

[0184] (S)-2-((R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-hydroxypropyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3g

[0185] 3f (169 mg, 0.34 mmol) was dissolved in tetrahydrofuran (8 mL), and ethanol (8 mL), sodium borohydride (77 mg, 2.05 mmol), and lithium chloride (94 mg, 2.22 mmol) were added. The mixture was stirred at 28 °C for 2.5 hours, and the reaction was quenched dropwise with saturated ammonium chloride (5 mL). The solution was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (10 mL), washed with water (10 mL), washed with saturated sodium chloride (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system A to give 3 g (125 mg, 78.4%) of the title product.

[0186] MS m / z (ESI): 467.2 [M+1].

[0187] Step 5

[0188] (S)-2-((R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(phenylthio)propyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3h

[0189] 3 g (120 mg, 0.26 mmol) was dissolved in toluene (5 mL), and diphenyl disulfide (168 mg, 0.77 mmol) and tributylphosphine (156 mg, 0.77 mmol) were added. The reaction system was heated to 80 °C and stirred for 12 hours under a nitrogen atmosphere. The solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using eluent system A to give the title product 3 h (135 mg, 93.9%).

[0190] MS m / z (ESI): 559.2 [M+1].

[0191] Step 6

[0192] (S)-2-((R)-2-amino-3-(phenylthio)propyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3i

[0193] 3h (135 mg, 0.24 mmol) was dissolved in dichloromethane (4 mL), and diethylamine (4 mL) was added. The mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure to remove the organic solvent, yielding the title product 3i (81.6 mg). The product was directly proceeded to the next step without purification.

[0194] Step 7

[0195] (S)-2-((R)-4-(phenylthio)-3-((4-aminosulfonyl-2-((trifluoromethyl)sulfonyl)phenyl)amino)butyl)

[0196] 3k Pyrrolidine-1-carboxylic acid tert-butyl ester

[0197] Crude product 3i (81.6 mg, 0.24 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 4-fluoro-3-(trifluoromethylsulfonyl)benzenesulfonamide 3j (82 mg, 0.27 mmol) and N,N-diisopropylethylamine (314 mg, 2.4 mmol) were added. The reaction system was heated to 50 °C and stirred for 12 hours under a nitrogen atmosphere. The reaction was quenched with water (10 mL), extracted with ethyl acetate (5 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system A to give the title product 3k (105 mg, 69.3%).

[0198] MS m / z (ESI): 622.1 [M-1].

[0199] Step 8

[0200] (S)-2-((R)-2-((4-(N-(4-(4-(((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)benzoyl)aminosulfonyl)-2-((trifluoromethyl)sulfonyl)phenyl)amino)-3-(phenylthio)propyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3m

[0201] Dissolve 3k (30 mg, 0.048 mmol) in dichloromethane (3 mL), add 3l (24 mg, 0.058 mmol, synthesized according to the method provided for intermediate 40 on page 79 of the specification of patent "CN103153954 B"), then add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (19 mg, 0.096 mmol) and 4-dimethylaminopyridine (12 mg, 0.096 mmol) in sequence. After the addition is complete, stir at room temperature for 4 hours under nitrogen atmosphere. Then add 3l (12 mg, 0.038 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (19 mg, 0.096 mmol) and 4-dimethylaminopyridine (12 mg, 0.096 mmol) in addition. After the addition is complete, stir at room temperature overnight. The reaction solution was washed with water (5 mL), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by thin-layer chromatography using solvent system A to give the title product 3m (30 mg, yield: 60.7%).

[0202] MS m / z (ESI): 1027.2 [M+1].

[0203] Step 9

[0204] 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-(phenylthio)-3-((S)-pyrrolidine-2-yl)propyl-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide3n

[0205] 3M (30 mg, 0.029 mmol) was dissolved in dichloromethane (2 mL), and a 4M solution of 1,4-dioxane in hydrogen chloride (2 mL) was added. The mixture was stirred at room temperature for 20 minutes, and the organic solvent was removed by concentration under reduced pressure. The residue was dissolved in dichloromethane and then concentrated to dryness under reduced pressure. This process was repeated three times. The resulting residue was purified by high performance liquid chromatography (HPLC) (separation conditions: column: Sharpsil-TPrep C18; mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding fractions were collected and concentrated under reduced pressure to give the title product 3n (25 mg, yield: 92.3%).

[0206] MS m / z (ESI): 927.1 [M+1].

[0207] 1 H NMR(500MHz,CD3OD)δ8.24(d,1H),8.05-8.01(m,1H),7.82(d,2H),7.60(d,1H),7.45-7.39(m,3H),7.37(d,2H),7.34-7.28(m ,3H),7.23(t,2H),7.20-7.14(m,2H),6.80(d,2H),6.72(d,1H),4.42(d,1H),3.97(br,1H),3.84-3.74(m,1H),3.27-3.21(m,2 H),3.19-3.07(m,3H),3.06-2.97(m,1H),2.69-2.61(m,1H),2.55-2.48(m,1H),2.16-2.00(m,5H),1.98-1.89(m,2H),1.88-1 .80(m,1H),1.78-1.69(m,1H),1.63-1.57(m,2H),1.54-1.49(m,1H),1.26-1.20(m,1H),1.15-1.04(m,1H),1.01-0.94(m,1H).

[0208] Step 10

[0209] 4-(4-((R)-(4'-chloro-[1,1'-biphenyl]-2-yl)(hydroxy)methyl)piperidin-1-yl)-N-((4-(((R)-1-((S)-1-(2-hydroxyethyl)pyrrolidine-2-yl)-3-(phenylthio)propyl-2-yl)amino)-3-((trifluoromethyl)sulfonyl)phenyl)sulfonyl)

[0210] Benzamide 3

[0211] 3n (15 mg, 0.016 mmol) was dissolved in acetonitrile (2 mL), and triethylamine (16 mg, 0.16 mmol) and bromoethanol (20 mg, 0.16 mmol) were added sequentially. The reaction system was heated to 70 °C for 12 hours under a nitrogen atmosphere. The product was purified by high performance liquid chromatography (separation conditions: column: Sharpsil-T Prep C18; mobile phase: ammonium bicarbonate, water, acetonitrile). The corresponding fractions were collected and concentrated under reduced pressure to give title product 3 (10 mg, yield: 63.4%).

[0212] MS m / z(ESI): 971.2 [M+1].

[0213] 1 H NMR(500MHz,CD3OD)δ8.25(d,1H),8.06(dd,1H),7.80(d,2H),7.61(dd,1H),7.49-7.35(m,5H),7.35-7.28(m,3H),7.24(d d,2H),7.20-7.14(m,2H),6.79(t,3H),4.42(d,1H),4.02(t,1H),3.84-3.70(m,3H),3.61(d,2H),3.28-3.14(m,2H),3.06( br,1H),2.96(br,1H),2.71-2.59(m,1H),2.56-2.45(m,1H),2.45-2.35(m,1H),2.32-2.22(m,1H),2.10-1.97(m,4H),1.96 -1.86(m,1H),1.85-1.69(m,2H),1.60(t,2H),1.26-1.18(m,1H),1.06-1.08(m,1H),1.01-0.92(m,1H),0.92-1.86(m,1H).

[0214] Example 2: Preparation of compound 3-TMS

[0215]

[0216] first step

[0217] 3n (6 g, 6.47 mmol) was dissolved in DCM (120 mL), and triethylamine (3.22 g, 32.35 mmol) was added. The mixture was cooled to 0 °C in an ice-water bath. Trimethylsilyltrifluoromethanesulfonate (7.2 g, 32.35 mmol) was slowly added dropwise to the reaction solution, and the mixture was stirred in an ice-water bath for 2 h. The reaction was quenched by adding methanol (12 mL) dropwise in an ice-water bath, and the mixture was stirred in an ice-water bath for 10 min. The solvent was removed under reduced pressure to obtain 3016.5 g of crude product, which was then proceeded to the next step without purification.

[0218] MS-ESI: m / z 999.1 [M+1] + .

[0219] Step 2

[0220] 3O (16.5 g, calculated as 6.47 mmol) was dissolved in acetonitrile (90 mL), and triethylamine (7.86 g, 77.64 mmol) and bromoethanol (9.7 g, 77.64 mmol) were added. The mixture was purged with nitrogen three times and heated to 70 °C overnight (approximately 16 h). The solvent was removed under reduced pressure, and EA (120 mL) and water (120 mL) were added. The mixture was stirred and separated. The aqueous phase was extracted with EA (60 mL × 2). The organic phases were combined, washed with water (120 mL), washed with saturated brine (120 mL), dried, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM:CH3OH = 20:1-10:1) to give compound 3-TMS 5.03 g, with a two-step yield of 74.5% and a purity of 95.9%.

[0221] MS-ESI: m / z 1043.05 [M+1] + .

[0222] 1 H NMR(400MHz, CDCl3)δ8.29(s,1H),8.05(s,1H),7.74(d,2H),7.55(t,1H),7.43–7.33(m ,3H),7.29(dd,3H),7.24–7.06(m,6H),6.97(d,1H),6.72(d,2H),6.63(d,1H),4.54(d,1 H),3.98(s,1H),3.82–3.46(m,5H),3.02(d,5H),2.82–2.44(m,4H),2.26(dd,1H),2.07 (d,1H),1.90(dd,3H),1.74(d,2H),1.58–1.41(m,1H),1.40–1.15(m,4H),-0.06(s,9H).

[0223] Example 3: Preparation of compound 1-Z00

[0224]

[0225]

[0226] first step

[0227] DMF (4 mL), methyliminodiacetic acid (218 mg, 1.48 mmol), and DCC (245 mg, 1.18 mmol) were added sequentially to a reaction flask. The mixture was cooled in an ice-water bath. Docetaxel (400 mg, 0.495 mmol) and DMAP (60 mg, 0.495 mmol) were then added, and the mixture was stirred at room temperature for 1.5 h. DCC (102 mg, 0.495 mmol) was added, and the mixture was stirred at room temperature for another 1 h. The reaction was stopped once it was confirmed to be complete. The mixture was filtered, and the filter cake was washed with ethyl acetate (15 mL). The filtrate was concentrated under reduced pressure to remove the ethyl acetate. The remaining mother liquor was purified by HPLC to obtain 86 mg of compound 1-Z01, yield: 19%.

[0228] MS-ESI: m / z 937.4 [M+H] + .

[0229] Step 2

[0230] Under a nitrogen atmosphere, 1-Z01 (1.59 g, 1.70 mmol) and PyBOP (1.70 g, 3.27 mmol) were dissolved in anhydrous DMF (28.0 mL) and stirred until homogeneous. Then, dendritic polymer 1 (2.00 g, 4.25 x 10⁻⁶) was added. -2 Anhydrous DMF (28.0 mL) solution containing 3.27 mmol of DIPEA (0.42 g, 3.27 mmol) and DIPEA (0.42 g, 3.27 mmol) was added dropwise to the above reaction solution, and the reaction was allowed to proceed for 2 hours. The reaction solution was then diluted with acetonitrile (56.0 mL) and filtered through ultrafiltration (10 kDa). The apparatus was used to purify the crude product by ultrafiltration with acetonitrile, yielding 2.70 g of crude product. This crude product was dissolved by vortexing in 150 mL of pure water, filtered through a 0.22 μm membrane, and lyophilized to give 2.40 g of compound 1-Z00, with a yield of 85%.

[0231] 1 1H NMR indicates 25DTX / dendritic structure. The actual molecular weight is approximately 66.4 kDa (30.39% by weight DTX).

[0232] 1H NMR (400MHz, CD3OD) δ (ppm) 8.05-8.24 (m, 64H), 7.11-7.78 (m, 272H), 5.98-6.25 (m, 25H), 5.55-5.71 (m, 29H), 5.19-5.49 (m, 84H), 4.93-5.0 7(s,47H),4.10-4.64(m,162H),3.40-4.08(m,3200H),3.33-3.37(S,9 6H), 3.02-3.23(m,126H), 2.14-2.63(m,271H), 0.99-2.10(m,1109H).

[0233] Example 4: Preparation of compound 1-Z00-J

[0234]

[0235] first step

[0236] Compound 1-Z01-J-1 (7 g, 29.5 mmol) was dissolved in methanol (150 mL), and formaldehyde solution (15 mL) and NaBH(AcO)3 (63 g, 295 mmol) were added. The reaction was carried out at room temperature for 1 hour. After concentration, the solution was purified by column chromatography to give 5.9 g of compound 1-Z01-JP, with a yield of 75%.

[0237] 1 H-NMR (400MHz, DMSO-d6) δ7.36-7.32(m,5H),5.09(s,2H),3.12(t,J=7.2Hz,1H),2.5-2.44(m,2H),2.38(s,6H),1.87-1.84(m,2H). MS-ESI:m / z 266.2[M+H] + .

[0238] Step 2

[0239] Compound 1-Z01-JP (1.44 g, 5.45 mmol) and docetaxel (4 g, 4.96 mmol) were dissolved in dichloromethane (50 mL), and EDCI (1.04 g, 5.45 mmol) and DMAP (665 mg, 5.45 mmol) were added. The mixture was stirred overnight at room temperature. The solution was quenched with water, extracted with dichloromethane, concentrated, and purified by column chromatography to give 5 g of compound 1-Z01-J-Bn in 95% yield.

[0240] MS-ESI: m / z 1055.5 [M+H] + .

[0241] Step 3

[0242] Compound 1-Z01-J-Bn (5.75 g, 5.46 mmol) was dissolved in tetrahydrofuran (60 mL), and Pd / C (600 mg, 10 wt.%) was added. Hydrogen was purged three times, and the reaction was carried out overnight at room temperature. After filtration and concentration, the solution was purified by column chromatography to obtain 3.2 g of compound 1-Z01-J, with a yield of 57%.

[0243] MS-ESI: m / z 965.3 [M+H] + .

[0244] Step 4

[0245] Under a nitrogen atmosphere, compound 1-Z01-J (819 mg, 849.5 μmol) and PyBOP (853 mg, 1640 μmol) were dissolved in anhydrous N,N-dimethylformamide (14 mL). A solution of dendritic polymer 1 (1000 mg, 21.24 μmol) and DIPEA (212 mg, 1640 μmol) in anhydrous N,N-dimethylformamide (14 mL) was added dropwise, and the reaction was allowed to proceed for 2 hours. The reaction solution was then diluted with acetonitrile (28 mL) and filtered through ultrafiltration (10 kDa). The apparatus was used to purify the crude product by ultrafiltration with acetonitrile, yielding 1.42 g of crude product. This crude product was dissolved in 100 mL of pure water, filtered through a 0.22 μm membrane, and lyophilized to give 1.42 g of compound 1-Z00-J, with a yield of 95%.

[0246] 1 1H NMR indicates 25DTX / dendritic structure. The actual molecular weight is approximately 67.1 kDa (30.0% wt% DTX).

[0247] 1 H NMR (400MHz, CD3OD) δ7.15-8.31(m,405H),6.00-6.31(m,25H),5.57-5.73(m,25H),5.20-5.54(m,83H),4.96-5.08(m,35H),4. 10-4.68(m,141H),3.40-4.08(m,3200H),3.33-3.37(s,103H),3.07-3.24(m,92H),2.14-2.69(m,260H),1.00-2.10(m,1183H).

[0248] Example 5: Preparation of compound 1-Z00-K

[0249]

[0250] first step

[0251] Compound 1-Z01-J01 (5.0 g, 21.09 mmol, from Jier Biochemical (Shanghai) Co., Ltd.) was dissolved in 100 mL of MeOH and stirred. Then, an aqueous formaldehyde solution (37% wt, 10 mL) was added. NaBH(AcO)3 (22.3 g, 105.4 mmol) was added in portions under ice bath cooling. After the addition was complete, the reaction mixture was reacted at room temperature for 18 hours. Upon completion of the reaction, the reaction solution was concentrated under reduced pressure. The residue was purified by column chromatography to give 3.2 g of compound 1-Z01-K-2, with a yield of 57%.

[0252] MS-ESI: m / z 266.1 [M+H] + .

[0253] 1 H-NMR: (CDCl3, 400MHz) δ7.36-7.27 (m, 5H), 5.18 (dd, J1=20.0Hz, J2=12.0Hz ,2H), 3.42-3.36(m,2H), 2.48-2.46(m,2H),2.45(s,6H),2.05-2.00(m,2H).

[0254] Step 2

[0255] Under a nitrogen atmosphere, compound 1-Z01-K-2 (1.0 g, 1.23 mmol), docetaxel (1.0 g, 1.23 mmol), EDCI (261 mg, 1.36 mmol), and DMAP (166 mg, 1.36 mmol) were added to a reaction flask, followed by the addition of anhydrous DMF (10 mL). The mixture was stirred overnight at room temperature. Saturated sodium bicarbonate solution and ethyl acetate were added, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to give 1.021 g of compound 1-Z01-K-3, in 71% yield.

[0256] MS-ESI: m / z 1055.4 [M+H] + .

[0257] Step 3

[0258] The starting material 1-Z01-K-3 (1.0 g, 0.947 mmol) and wet palladium on carbon (10% wt, 200 mg) were added to a reaction flask, followed by the addition of tetrahydrofuran (20 mL). The mixture was purged with hydrogen three times and reacted at room temperature for 18 hours. The reaction mixture was filtered through diatomaceous earth, washed with ethyl acetate, and the filtrate was concentrated and dried under vacuum to give 736 mg of compound 1-Z01-K, in 80% yield.

[0259] MS-ESI: m / z 965.3 [M+H] + .

[0260] Step 4

[0261] Under a nitrogen atmosphere, compound 1-Z01-K (162 mg, 0.168 mmol) and PyBOP (168 mg, 0.323 mmol) were dissolved and stirred in anhydrous DMF (6.0 mL) until homogeneous. Then, dendritic polymer 1 (200 mg, 4.2 x 10⁻⁶ mmol) was added. -3 An anhydrous DMF solution of 6.0 mL containing 0.323 mmol of DIPEA (42 mg, 0.323 mmol) and 0.323 mmol of DIFEA (42 mg, 0.323 mmol) was added dropwise to the above reaction solution, and the reaction was allowed to proceed for 2 hours. The reaction solution was then diluted with acetonitrile and filtered through ultrafiltration (10 kDa). The apparatus was used to purify the crude product by ultrafiltration with acetonitrile, yielding 0.215 g of crude product. This crude product was dissolved in pure water (50 mL), filtered through a 0.22 μm membrane, and lyophilized to give 0.21 g of compound 1-Z00-K, with a yield of 76%.

[0262] 1 1H NMR indicates 24DTX / dendritic structure. The actual molecular weight is approximately 66.1 kDa (29.30% wt% DTX).

[0263] 1 H NMR (400MHz, CD3OD) δ (ppm) 8.00-8.23 (m, 55H), 7.11-7.78 (m, 239H), 5.95-6.30 (m, 24H), 5.51-5.75 (m, 24H), 5.15-5.45 (m, 72H), 4.93-5.07 (s,19H),4.10-4.64(m,157H),3.40-4.08(m,3200H),3.33-3.37(s,10 2H), 2.93-3.23(m,167H), 2.12-2.63(m,212H), 0.99-2.10(m,1145H).

[0264] Example 6: Preparation of compound 1-Z00-L

[0265]

[0266] first step

[0267] Compound 1-Z01-J-1 (1.0 g, 4.2 mmol, Shanghai Bide Pharmaceutical) was dissolved in MeOH (20 mL) with stirring, followed by the addition of acetaldehyde aqueous solution (40% wt, 2 mL). NaBH(AcO)3 (4.472 g, 21.0 mmol) was added under ice bath conditions, and the reaction was carried out at room temperature for 20 hours. The mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to give 733 mg of compound 1-Z01-L-1, with a yield of 59.6%.

[0268] MS-ESI: m / z 294.1 [M+H] + .

[0269] 1 H-NMR: (CDCl3, 400MHz) δ7.35~7.28 (m, 5H), 5.10 (dd, J1=18Hz, J2=12.4Hz, 2H), 4.15 (brs 1H), 3.56 (t, J=6.8Hz 1H), 3.27~3.22(m,2H), 3.07-3.02(m,2H), 2.86~2.69(m,2H), 2.05~2.00(m,2H), 1.30(t,J=7.6Hz,6H).

[0270] Step 2

[0271] Compound 1-Z01-L-1 (399 mg, 1.36 mmol), docetaxel (1.0 g, 1.23 mmol), EDCI (261 mg, 1.36 mmol), and DMAP (166 mg, 1.36 mmol) were added to a reaction flask, followed by the addition of anhydrous DMF (10 mL). The mixture was stirred overnight at room temperature. Saturated sodium bicarbonate solution and ethyl acetate were added, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography to give 798 mg of compound 1-Z01-L-2, in 54% yield.

[0272] MS-ESI: m / z 1083.4 [M+H] + .

[0273] Step 3

[0274] Compound 1-Z01-L-2 (790 mg, 0.729 mmol) and wet palladium on carbon (10% wt, 158 mg) were added to a reaction flask, followed by the addition of tetrahydrofuran (16 mL). The mixture was purged with hydrogen three times and stirred at room temperature for 18 hours. The reaction mixture was filtered through diatomaceous earth, washed with ethyl acetate, and the filtrate was concentrated and dried to give 616 mg of compound 1-Z01-L, with a yield of 85%.

[0275] MS-ESI: m / z 993.4 [M+H] + .

[0276] Step 4

[0277] Under a nitrogen atmosphere, compound 1-Z01-L (530 mg, 0.53 mmol) and PyBOP (537 mg, 1.03 mmol) were dissolved in anhydrous DMF (9.0 mL) and stirred until homogeneous. Then, dendritic polymer 1 (629 mg, 13.4 x 10⁻⁶ mmol) was added.-3 An anhydrous DMF solution of 1.03 mmol (1.03 mg) and DIPEA (133 mg, 1.03 mmol) in 9.0 mL was added dropwise to the above reaction solution, and the reaction was allowed to proceed for 2 hours. The reaction solution was then diluted with acetonitrile and filtered through ultrafiltration (10 kDa). The apparatus was used to purify the product by ultrafiltration with acetonitrile, yielding 0.66 g of crude product. This crude product was dissolved in pure water (100 mL), filtered through a 0.22 μm filter membrane, and lyophilized to obtain 0.65 g of compound 1-Z00-L, with a yield of 71%.

[0278] 1 1H NMR indicates 26DTX / dendritic structure. The actual molecular weight is approximately 68.8 kDa (30.51% by weight DTX).

[0279] 1 H NMR (400MHz, CD3OD) δ (ppm) 7.20-8.25 (m, 288H), 5.98-6.30 (m, 26H), 5.52-5.78 (m, 28H), 5.15-5.45 (m, 84H), 4.92-5.08 (s, 31H), 4.10-4.64(m,212H),3.40-4.08(m,3200H),3.33-3.37(S,102H),2.93-3.23(m,172H),2.12-2.63(m,262H),0.99-2.10(m,1186H).

[0280] Example 7: Preparation of compound 1-Z00-M

[0281]

[0282]

[0283] first step

[0284] Compound 1-Z01-J-1 (5.1 g, 21.496 mmol, Shanghai Bide Pharmaceutical) was dissolved in 100 mL of MeOH and stirred. Then, propionaldehyde (6.3 g, 108.47 mmol) was added, and the mixture was cooled in an ice bath before adding NaBH(AcO)3 (22.9 g, 108.02 mmol). After the addition was complete, the reaction mixture was reacted at room temperature for 20 hours. The reaction solution was concentrated under reduced pressure to remove methanol. The residue was mixed with water, stirred, and filtered to remove insoluble matter. The filtrate was purified by column chromatography to give 4.85 g of compound 1-Z01-M-1, yield: 68.7%.

[0285] MS-ESI: m / z 322.2 [M+H] + .

[0286] 1H-NMR (CDCl3, 400MHz) δ7.37-7.26(m,5H), 5.12(s,2H), 3.53(brs 1H),3.04-2.72(m,7H),2.09-1.98(m,2H),1.79-1.67(m,4H),0.97-0.93(m,6H).

[0287] Step 2

[0288] Under a nitrogen atmosphere, compound 1-Z01-M-1 (367 mg, 1.1 mmol), docetaxel (801 g, 0.991 mmol), EDCI (212 mg, 1.1 mmol), and DMAP (136 mg, 1.1 mmol) were added to a reaction flask, followed by the addition of 10 mL of anhydrous DMF. The mixture was stirred overnight at room temperature. Saturated sodium bicarbonate solution and ethyl acetate were added, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography to give 732 mg of compound 1-Z01-M-2, in 66% yield.

[0289] MS-ESI: m / z 1111.4 [M+H] + .

[0290] Step 3

[0291] Compound 1-Z01-M-2 (732 mg, 0.658 mmol) and wet palladium on carbon (10% wt, 133 mg) were added to a reaction flask, followed by tetrahydrofuran (18 mL). The mixture was purged with hydrogen three times and stirred at room temperature for 18 hours. The reaction mixture was filtered through diatomaceous earth, washed with ethyl acetate, and the filtrate was concentrated under reduced pressure to give 669 mg of compound 1-Z01-M, in 99% yield.

[0292] MS-ESI: m / z 1021.4 [M+H] + .

[0293] Step 4

[0294] Under a nitrogen atmosphere, compound 1-Z01-M (500 mg, 0.49 mmol) and PyBOP (490 mg, 0.943 mmol) were dissolved and stirred in anhydrous DMF (7.5 mL) until homogeneous. Then, dendritic polymer 1 (543 mg, 12.25 x 10⁻⁶ mmol) was added. -3 An anhydrous DMF solution of 7.5 mL containing 122 mg (0.943 mmol) and DIPEA (122 mg, 0.943 mmol) was added dropwise to the above reaction solution, and the reaction was allowed to proceed for 2 hours. The reaction solution was then diluted with acetonitrile and filtered through ultrafiltration (10 kDa). The apparatus was used to purify the product by ultrafiltration with acetonitrile, yielding 0.64 g of crude product. This crude product was dissolved in pure water (100 mL), filtered through a 0.22 μm membrane, and lyophilized to obtain 0.63 g of compound 1-Z00-M, with a yield of 83%.

[0295] 1 1H NMR indicates 21DTX / dendritic structure. The actual molecular weight is approximately 61.7 kDa (27.4% wt% DTX).

[0296] 1 H NMR (400MHz, CD3OD) δ (ppm) 8.00-8.22 (m, 45H), 7.09-7.81 (m, 198H), 5.95-6.30 (m, 21H), 5.51-5.75 (m, 20H), 5.13-5.45 (m, 62H), 4.93-5.0 7(s,17H),4.10-4.64(m,175H),3.40-4.08(m,3000H),3.33-3.37(S,9 1H), 2.93-3.23(m,127H), 2.12-2.63(m,217H), 0.79-2.10(m,1396H).

[0297] Example 8: Preparation of compound 1-Z00-Q

[0298]

[0299] first step

[0300] Compound 1-Z01-J-1 (5 g, 21.1 mmol, Shanghai Bide Pharmaceutical) was dissolved in DCM (50 mL). Under a nitrogen atmosphere, triethylamine (6.41 g, 63.3 mmol) was added, and the mixture was cooled in an ice-water bath. Acetic anhydride (2.37 g, 23.2 mmol) was added dropwise, and the reaction was allowed to proceed overnight at room temperature. The reaction solution was washed with water, and the aqueous phases were combined. The pH was adjusted to 1 with concentrated hydrochloric acid in an ice-water bath. The mixture was extracted three times with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 5.8 g of a white solid. A mixture of ethyl acetate and petroleum ether was added and stirred overnight. The mixture was filtered to give 4.9 g of compound 1-Z01-Q-1, yield: 83%. This was used directly in the next step.

[0301] MS-ESI: m / z 280.2 [M+H] + .

[0302] 1H NMR(400MHz,DMSO-d6)δ12.62(br,1H),8.14(d,J=8Hz,1H),7.40-7.31(m,5H),5.09(s,2H) ,4.24-4.18(m,1H),2.48-2.38(m,2H),2.05-2.00(m,1H),1.98-1.77(m,1H),1.97(s,3H).

[0303] Step 2

[0304] Compound 1-Z01-Q-1 (377 mg, 1.350 mmol), docetaxel (1 g, 1.239 mmol), and DMAP (166 mg, 1.363 mmol) were weighed into a reaction flask. Under a nitrogen atmosphere and in an ice-water bath, 10 mL of dry DMF and 261 mg, 1.363 mmol were added. After the addition was complete, the reaction mixture was allowed to react overnight at room temperature. Ethyl acetate was added to the reaction solution, and the organic phase was washed successively with saturated sodium bicarbonate solution and saturated NaCl solution. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography to give 805 mg of compound 1-Z01-Q-2, yield: 60.8%.

[0305] MS-ESI: m / z 1069.5 [M+H] + .

[0306] Step 3

[0307] Compound 1-Z01-Q-2 (1.1 g, 1.029 mmol) was dissolved in 20 mL of THF, and wet palladium on carbon (10% wt, 220 mg) was added. The mixture was purged three times with hydrogen, and then stirred overnight at room temperature under a hydrogen atmosphere. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure and dried under vacuum to give 962 mg of compound 1-Z01-Q, yield: 95.5%.

[0308] MS-ESI: m / z 979.4 [M+H] + .

[0309] Step 4

[0310] Under a nitrogen atmosphere, compound 1-Z01-Q (500 mg, 0.511 mmol) and PyBOP (513 mg, 0.986 mmol) were dissolved and stirred in anhydrous DMF (7.9 mL) until homogeneous. Then, dendritic polymer 1 (567 mg, 12.8 x 10⁻⁶ mmol) was added. -3 An anhydrous DMF solution of 7.9 mL containing 127 mg (1 mmol) and 0.986 mol (127 mg, 0.986 mol) was added dropwise to the above reaction solution, and the reaction was allowed to proceed for 2 hours. The reaction solution was then diluted with acetonitrile and filtered through ultrafiltration (10 kDa). The apparatus was used to purify the product by ultrafiltration with acetonitrile, yielding 0.625 g of crude product. This crude product was dissolved in pure water (100 mL), filtered through a 0.22 μm filter membrane, and lyophilized to give 0.62 g of compound 1-Z00-Q, with a yield of 74%.

[0311] 1 1H NMR indicates 26DTX / dendritic structure. The actual molecular weight is approximately 65.7 kDa (31.96% by weight DTX).

[0312] 1 H NMR (400MHz, CD3OD) δ8.00-8.22(m,55H),7.09-7.81(m,253H),5.95-6.31(m,26H),5.51-5.77(m,28H),5.13-5.45(m,91H),4.92-5.08(s ,41H),4.10-4.64(m,225H),3.40-4.08(m,3000H),3.33-3.37(s,91H),2.93-3.23(m,102H),2.12-2.63(m,310H),0.96-2.10(m,1119H).

[0313] Example 9: Preparation of compound 3-L00

[0314]

[0315] first step

[0316] Compound 1-Z01-J01 (5.0 g, 18.3 mmol) was dissolved in methanol (100 mL), cooled to 0 °C, and then an aqueous formaldehyde solution (10 mL, 40 wt.%) and NaBH(OAc)3 (20.0 g, 91.6 mmol) were added. The mixture was reacted overnight at room temperature. After concentration, the solution was purified by column chromatography to give 3.4 g of compound 1-Z01-J02, yield: 61%.

[0317] MS-ESI: m / z 266.1 [M+H] + .

[0318] Step 2

[0319] Compound 1-Z01-J02 (3.4 g, 12.8 mmol), trimethylsilylethanol (3.0 g, 26.0 mmol), DMAP (780 mg, 6.4 mmol), and N,N-diisopropylethylamine (6.6 g, 51.2 mmol) were dissolved in anhydrous tetrahydrofuran (30 mL). The solution was cooled to 0 °C, and HATU (6.3 g, 16.6 mmol) was added. The reaction was carried out at room temperature for 16 hours. The reaction was quenched with water (50 mL), extracted with ethyl acetate (50 mL × 3), concentrated, and purified by column chromatography to give 4.0 g of compound 1-Z01-J03, yield: 85%.

[0320] MS-ESI: m / z 366.2 [M+H] + .

[0321] Step 3

[0322] Compound 1-Z01-J03 (4.0 g, 10.9 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL), Pd / C (200 mg, 10 wt.%) was added, and the mixture was purged three times with hydrogen. The reaction was carried out at room temperature for 16 hours. After filtration and concentration, 3.6 g of crude product was obtained. Anhydrous tetrahydrofuran (30 mL) was added, and the mixture was stirred for 30 minutes. After filtration, 1.6 g of compound 1-Z01-J-TMS was obtained, with a yield of 45%.

[0323] MS-ESI: m / z 276.1 [M+H] + .

[0324] 1 H NMR (400MHz, CDCl3) δ4.14-4.18(m,2H),3.44-3.47(m,1H),2.87(s,6H),2.66-2.70(m,2H),2.03-2.12(m,2H),0.96-0.99(m,2H),0.04(s,9H).

[0325] Step 4

[0326] Compounds 1-Z01-J-TMS (118 mg, 0.431 mmol), 3-TMS (300 mg, 0.287 mmol), and DMAP (17.5 mg, 0.216 mmol) were dissolved in anhydrous dichloromethane (10 mL), and DCC (88.8 mg, 0.431 mmol) was added. The mixture was reacted at room temperature for 16 hours. After filtration and concentration, the solution was purified by column chromatography to obtain 373 mg of compound 3-L00-1, yield: 98%.

[0327] MS-ESI: m / z 650.8 [1 / 2M+H] + .

[0328] Step 5

[0329] Compound 3-L00-1 (373 mg, 0.288 mmol) was dissolved in anhydrous tetrahydrofuran (3 mL), and TBAF (2.9 mL, 2.88 mmol, 1.0 mol / L THF solution) was added. The reaction was carried out at room temperature for 2 hours. The reaction was quenched with water (10 mL), extracted with ethyl acetate (10 mL × 3), concentrated, and purified by column chromatography to give 238 mg of compound 3-L00-2, yield: 73%.

[0330] MS-ESI: m / z 564.5 [1 / 2M+H] + .

[0331] Step 6

[0332] Under a nitrogen atmosphere, compound 3-L00-2 (0.390 g, 0.346 mmol) and PyBOP (0.216 g, 0.415 mmol) were dissolved in anhydrous N,N-dimethylformamide (5.4 mL), and a solution of dendritic polymer 1-PEG2K (0.535 g, 6.92 μmol, synthesized according to the literature Journal of Controlled Release, 2011, 152, 241-248) and NMM (0.139 g, 1.38 mmol) in anhydrous N,N-dimethylformamide (5.4 mL) was added dropwise. The reaction was carried out at room temperature for 2 hours. The reaction solution was diluted with acetonitrile (11 mL) and then filtered through ultrafiltration (10 kDa). The apparatus was used to purify the crude product by ultrafiltration with acetonitrile (0.55 L, 25 V) to obtain 0.625 g of crude product. This crude product was dissolved in pure water, filtered through a 0.22 μm membrane, and lyophilized to give 0.62 g of compound 3-L00, yield: 89%.

[0333] 1 1H NMR indicates 24 drug molecules / dendritic structures. The actual molecular weight is approximately 100.2 kDa (23.22% by weight, compound 3).

[0334] 1 H NMR(400MHz,CD3OD)δ6.40-8.51(m,498H),4.19-4.68(m,111H),3.40-4.10(m,5900H) ,3.33-3.36(s,106H),2.79-3.20(m,112H),2.22-2.74(m,246H),0.61-1.92(m,643H).

[0335] Example 10: Preparation of compound 3-M00

[0336]

[0337] first step

[0338] Compound 1-Z01-J-1 (7 g, 29.5 mmol) was dissolved in methanol (150 mL), and then an aqueous formaldehyde solution (15 mL, 40 wt.%) and NaBH(OAc)3 (63 g, 295 mmol) were added. The mixture was reacted at room temperature for 1 hour. After concentration, the solution was purified by column chromatography to give 5.9 g of compound 1-Z01-JP, yield: 75%.

[0339] MS-ESI: m / z 266.2 [M+H] + .

[0340] 1 H NMR (400MHz, DMSO-d6) δ7.36-7.32(m,5H),5.09(s,2H),3.12(t,J=7.2Hz,1H),2.5-2.44(m,2H),2.38(s,6H),1.87-1.84(m,2H).

[0341] Step 2

[0342] Compound 1-Z01-JP (4.2 g, 15.8 mmol), trimethylsilylethanol (2.24 g, 19 mmol), DMAP (0.193 g, 1.58 mmol), and N,N-diisopropylethylamine (6.1 g, 47.5 mmol) were dissolved in dichloromethane (42 mL), cooled to 0 °C, and HATU (7.2 g, 19 mmol) was added. The reaction was carried out at room temperature for 16 hours. The reaction was quenched with saturated sodium bicarbonate solution (150 mL), extracted with dichloromethane (50 mL × 3), concentrated, and purified by column chromatography to give 4.9 g of compound 1-Z01-K-1, with a yield of 86%.

[0343] MS-ESI: m / z 366.2 [M+H] + .

[0344] Step 3

[0345] Under a nitrogen atmosphere, compound 1-Z01-K-1 (900 mg, 2.46 mmol) was dissolved in tetrahydrofuran (18 mL), and Pd / C (180 mg, 10 wt.%) was added. The mixture was purged three times with hydrogen, and the reaction was carried out at room temperature for 16 hours. After filtration and concentration, 670 mg of compound 1-Z01-K-TMS was obtained, with a yield of 99%.

[0346] MS-ESI: m / z 276.1 [M+H] + .

[0347] 1H NMR (400MHz, CDCl3) δ4.22 (dd, J=10.0Hz, J=8.4Hz, 2H), 3.39 (t, J=7.2Hz, 1H), 2.65-2.58 (m, 1H), 2.49(s,6H),2.47-2.39(m,1H),2.02(dd,J=12.8Hz,J=6.8Hz,2H),1.04-1.00(m,2H),0.04(s,9H).

[0348] Step 4

[0349] Under a nitrogen atmosphere, compounds 3-TMS (800 mg, 0.767 mmol), 1-Z01-K-TMS (316 mg, 0.767 mmol), and DMAP (46 mg, 0.383 mmol) were dissolved in dichloromethane (16 mL), cooled to 0 °C, and DCC (237 mg, 1.15 mmol) was added. The mixture was reacted at room temperature for 18 hours. After filtration and concentration, the solution was purified by column chromatography to give 1.036 g of compound 3-M00-1, yield: 100%.

[0350] MS-ESI: m / z 650.8 [1 / 2M+H] + .

[0351] Step 5

[0352] Under a nitrogen atmosphere, compound 3-M00-1 (1.0 g, 0.768 mmol) was dissolved in tetrahydrofuran (10 mL), cooled to 0 °C, and TBAF (7.7 mL, 7.68 mmol, 1.0 mol / L THF solution) was added. The reaction was carried out at room temperature for 1 hour. The reaction was quenched with water (30 mL), extracted with ethyl acetate (50 mL × 3), concentrated, and purified by column chromatography to give 670 mg of compound 3-M00-2, yield: 77%.

[0353] MS-ESI: m / z 1128.3 [M+H] + .

[0354] Step 6

[0355] Under a nitrogen atmosphere, compound 3-M00-2 (0.400 g, 0.355 mmol) and PyBOP (0.222 g, 0.426 mmol) were dissolved in anhydrous N,N-dimethylformamide (5.6 mL), and a solution of dendritic polymer 1-PEG2K (0.548 g, 7.10 μmol) and NMM (0.144 g, 1.42 mmol) in anhydrous N,N-dimethylformamide (5.6 mL) was added dropwise. The reaction was carried out at room temperature for 2 hours. The reaction solution was diluted with acetonitrile (11 mL) and then filtered through ultrafiltration (10 kDa). The apparatus was used to purify the crude product by ultrafiltration with acetonitrile (0.55 L, 25 V), yielding 0.614 g of crude product. This crude product was dissolved in pure water, filtered through a 0.22 μm membrane, and lyophilized to give 0.61 g of compound 3-M00, yield: 88%.

[0356] 1 1H NMR indicates 22 drug molecules / dendritic structures. The actual molecular weight is approximately 98.0 kDa (21.77% by weight, compound 3).

[0357] 1 H NMR(400MHz,CD3OD)δ6.40-8.52(m,440H), 4.13-4.70(m,137H),3.40-4.10(m,5900H) ,3.33-3.36(s,109H),2.79-3.25(m,99H),2.22-2.74(m,246H),0.61-1.92(m,622H).

[0358] Example 11: Preparation of compound 3-NO00

[0359]

[0360] first step

[0361] Compound 1-Z01-J01 (10.0 g, 36.6 mmol) was dissolved in methanol (200 mL), cooled to 0 °C, and then acetaldehyde aqueous solution (20 mL, 40 wt.%) was added. NaBH(OAc)3 (40.0 g, 183.2 mmol) was then added, and the mixture was reacted overnight at room temperature. After concentration, the solution was purified by column chromatography to give 5.4 g of compound 1-Z01-L02, yield: 50.4%.

[0362] MS-ESI: m / z 294.1 [M+H] + .

[0363] Step 2

[0364] Compound 1-Z01-L02 (500 mg, 1.7 mmol), trimethylsilylethanol (404 mg, 3.4 mmol), DMAP (104 mg, 0.85 mmol), and N,N-diisopropylethylamine (658 mg, 5.1 mmol) were dissolved in anhydrous tetrahydrofuran (10 mL), cooled to 0 °C, and HATU (840 mg, 2.2 mmol) was added. The reaction was carried out at room temperature for 16 hours. The reaction was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 3), concentrated, and purified by column chromatography to obtain 476 mg of compound 1-Z01-L03, yield: 71%.

[0365] MS-ESI: m / z 394.2 [M+H] + .

[0366] Step 3

[0367] Compound 1-Z01-L03 (476 mg, 1.2 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), Pd / C (50 mg, 10 wt.%) was added, and the mixture was purged with hydrogen three times. The reaction was carried out at room temperature for 16 hours. After filtration and concentration, 366 mg of compound 1-Z01-L-TMS was obtained, yield: 100%.

[0368] MS-ESI: m / z 304.2 [M+H] + .

[0369] 1 H NMR (400MHz, CD3OD) δ4.14-4.18(m,2H),3.58-3.60(m,1H),3.26-3.30(m,2H),3.06-3.13(m, 2H),2.66-2.77(m,2H),2.00-2.06(m,2H),1.33-1.38(m,6H),0.94-0.99(m,2H),0.04(s,9H).

[0370] Step 4

[0371] Compounds 1-Z01-L-TMS (366 mg, 1.21 mmol), 3-TMS (800 mg, 0.767 mmol), and DMAP (47.0 mg, 0.38 mmol) were dissolved in anhydrous dichloromethane (25 mL), and DCC (246.0 mg, 1.2 mmol) was added. The reaction was carried out at room temperature for 16 hours. After filtration and concentration, the solution was purified by column chromatography to give 1.0 g of compound 3-N00-1, yield: 98%.

[0372] MS-ESI: m / z 664.8 [1 / 2M+H] + .

[0373] Step 5

[0374] Compound 3-N00-1 (1.0 g, 0.75 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), and TBAF (7.5 mL, 7.5 mmol, 1.0 mol / L THF solution) was added. The reaction was carried out at room temperature for 2 hours. The reaction was quenched with water (20 mL), extracted with ethyl acetate (50 mL × 3), concentrated, and purified by column chromatography to give 740 mg of compound 3-N00-2, yield: 85%.

[0375] MS-ESI: m / z 578.7 [1 / 2M+H]+ .

[0376] Step 6

[0377] Under a nitrogen atmosphere, compound 3-N00-2 (0.400 g, 0.346 mmol) and PyBOP (0.215 g, 0.414 mmol) were dissolved in anhydrous N,N-dimethylformamide (5.6 mL). A solution of the dendritic polymer 1-PEG2K (0.535 g, 6.90 μmol) and NMM (0.139 g, 1.38 mmol) in anhydrous N,N-dimethylformamide (5.6 mL) was added dropwise, and the reaction was carried out at room temperature for 2 hours. The reaction solution was diluted with acetonitrile (11 mL) and then filtered through ultrafiltration (10 kDa). The apparatus was used to purify the product by ultrafiltration with acetonitrile (0.56 L, 25 V) to obtain 0.631 g of crude product. This crude product was dissolved in pure water, filtered through a 0.22 μm membrane, and lyophilized to give 0.625 g of compound 3-N00, yield: 91%.

[0378] 1 1H NMR indicates 23 drug molecules / dendritic structures. The actual molecular weight is approximately 99.8 kDa (22.36% by weight, compound 3).

[0379] 1 H NMR (400MHz, CD3OD) δ6.40-8.52(m,477H), 4.19-4.68(m,184H), 3.40-4.10(m,5900H) ,3.33-3.36(s,108H),2.79-3.20(m,101H),2.22-2.74(m,158H),0.61-1.92(m,726H).

[0380] Example 12: Preparation of compound 3-O00

[0381]

[0382] first step

[0383] Compound 1-Z01-J01 (5.0 g, 21.1 mmol) was dissolved in methanol (100 mL), cooled to 0 °C, and then propionaldehyde (6.1 g, 105.0 mmol) and NaBH(OAc)3 (31.1 g, 105.0 mmol) were added. The mixture was reacted overnight at room temperature. After concentration, the solution was purified by column chromatography to give 5.0 g of compound 1-Z01-M02, yield: 74%.

[0384] MS-ESI: m / z 322.1 [M+H] + .

[0385] Step 2

[0386] Compound 1-Z01-M02 (890 mg, 2.8 mmol), trimethylsilylethanol (393.3 mg, 3.3 mmol), DMAP (33.8 mg, 0.3 mmol), and N,N-diisopropylethylamine (1.1 g, 9.3 mmol) were dissolved in anhydrous dichloromethane (20 mL), cooled to 0 °C, and HATU (1.3 g, 3.3 mmol) was added. The reaction was carried out at room temperature for 16 hours. The reaction was quenched with water (40 mL), extracted with ethyl acetate (20 mL × 3), concentrated, and purified by column chromatography to give 1 g of compound 1-Z01-M03, yield: 86%.

[0387] MS-ESI: m / z 422.2 [M+H] + .

[0388] Step 3

[0389] Compound 1-Z01-M03 (1 g, 2.4 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL), and Pd / C (200 mg, 10 wt.%) was added. The mixture was purged with hydrogen three times, and the reaction was carried out at room temperature for 16 hours. After filtration and concentration, 766 mg of compound 1-Z01-M-TMS was obtained, yield: 100%.

[0390] MS-ESI: m / z 332.2 [M+H] + .

[0391] 1 H NMR (400MHz, CDCl3) δ4.10-4.14(m,2H),3.50-3.53(m,1H),2.84-3.04(m,4H),2.63 -2.69(m,2H),1.95-2.01(m,2H),1.69-1.77(m,4H),0.90-0.96(m,8H),0.04(s,9H).

[0392] Step 4

[0393] Compounds 1-Z01-M-TMS (381.7 mg, 1.2 mmol), 3-TMS (800 mg, 0.767 mmol), and DMAP (46.08 mg, 0.38 mmol) were dissolved in anhydrous dichloromethane (20 mL), and DCC (237.0 mg, 1.2 mmol) was added. The mixture was reacted at room temperature for 16 hours. After filtration and concentration, the solution was purified by column chromatography to obtain 890 mg of compound 3-O00-1, yield: 86%.

[0394] MS-ESI: m / z 678.8 [1 / 2M+H] + .

[0395] Step 5

[0396] Compound 3-O00-1 (890 mg, 0.66 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), and TBAF (6.6 mL, 6.6 mmol, 1.0 mol / L THF solution) was added. The reaction was carried out at room temperature for 3 hours. The reaction was quenched with water (20 mL), extracted with ethyl acetate (50 mL × 3), concentrated, and purified by column chromatography to give 810 mg of compound 3-O00-2, yield: 100%.

[0397] MS-ESI: m / z 592.8 [1 / 2M+H] + .

[0398] Step 6

[0399] Under a nitrogen atmosphere, compound 3-O00-2 (0.409 g, 0.346 mmol) and PyBOP (0.216 g, 0.415 mmol) were dissolved in anhydrous N,N-dimethylformamide (5 mL), and a solution of dendritic polymer 1-PEG2K (0.52 g, 6.8 μmol) and NMM (0.137 g, 1.36 mmol) in anhydrous N,N-dimethylformamide (5 mL) was added dropwise. The reaction was carried out at room temperature for 2 hours. The reaction solution was diluted with acetonitrile (10 mL) and then filtered through ultrafiltration (10 KD). The apparatus was used to purify the crude product by ultrafiltration with acetonitrile (0.5 L, 25 V) to obtain 0.605 g of crude product. This crude product was dissolved in pure water, filtered through a 0.22 μm membrane, and lyophilized to give 0.60 g of compound 3-O00, yield: 87%.

[0400] 1 1H NMR indicates 24 drug molecules / dendritic structures. The actual molecular weight is approximately 101.5 kDa (22.92% by weight, compound 3).

[0401] 1 H NMR(400MHz,CD3OD)δ6.40-8.51(m,496H), 4.19-4.68(m,126H),3.40-4.10(m,5900H) ,3.33-3.36(s,100H),2.79-3.20(m,136H),2.22-2.74(m,199H),0.61-1.92(m,683H).

[0402] Example 13: Preparation of compound 3-R00

[0403]

[0404] first step

[0405] Compound 1-Z01-J01 (5.01 g, 18.3 mmol) was dissolved in methanol (100 mL), cooled to 0 °C, and acetone (5.5 g, 94.7 mmol) and NaBH(OAc)3 (11.9 g, 56.13 mmol) were added. The reaction was carried out at room temperature for 2 hours. Formaldehyde aqueous solution (14 mL, 40 wt.%) and NaBH(OAc)3 (8.26 g, 38.96 mmol) were added, and the reaction was carried out at room temperature overnight. After concentration, the solution was purified by column chromatography to give 5 g of compound 1-Z01-PO2, yield: 93%.

[0406] MS-ESI: m / z 294.1 [M+H] + .

[0407] Step 2

[0408] Compound 1-Z01-PO2 (2.58 g, 8.8 mmol), trimethylsilylethanol (2 g, 16.9 mmol), DMAP (540 mg, 4.43 mmol), and N,N-diisopropylethylamine (2.98 g, 23.1 mmol) were dissolved in anhydrous tetrahydrofuran (20 mL), and HATU (3.38 g, 8.89 mmol) was added. The reaction was carried out at room temperature for 17 hours. The reaction was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 3), concentrated, and purified by column chromatography to give 3.09 g of compound 1-Z01-PO3, yield: 89%.

[0409] MS-ESI: m / z 394.2 [M+H] + .

[0410] Step 3

[0411] Compound 1-Z01-P03 (3 g, 7.62 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL), and Pd / C (152 mg, 10 wt.%) was added. The mixture was purged three times with hydrogen and reacted at room temperature for 15 hours. After filtration and concentration, 2.32 g of crude 1-Z01-P-TMS was obtained. Methyl tert-butyl ether (20 mL) was added, and the mixture was stirred for 30 minutes. After filtration, 0.92 g of compound 1-Z01-P-TMS was obtained, with a yield of 39%.

[0412] MS-ESI: m / z 304.2 [M+H] + .

[0413] 1HNMR(400MHz,DMSO-d6)δ4.05-4.10(m,2H),3.26-3.30(m,1H),3.12-3.15(m,1H),3.35-3.4 3(m,2H),2.31(s,3H),1.75-1.85(m,2H),1.03-1.07(m,6H),0.89-0.94(m,2H),0.01(s,9H).

[0414] Step 4

[0415] Compounds 1-Z01-P-TMS (379 mg, 1.25 mmol), 3-TMS (805 mg, 0.772 mmol), and DMAP (54.0 mg, 0.44 mmol) were dissolved in anhydrous dichloromethane (16 mL), and DCC (239.0 mg, 1.16 mmol) was added. The mixture was reacted at room temperature for 16 hours. After filtration and concentration, the solution was purified by column chromatography to obtain 863 mg of compound 3-R00-1, yield: 84%.

[0416] MS-ESI: m / z 664.8 [1 / 2M+H] + .

[0417] Step 5

[0418] Compound 3-R00-1 (847 mg, 0.64 mmol) was dissolved in anhydrous tetrahydrofuran (6 mL), and TBAF (6 mL, 6 mmol, 1.0 mol / L THF solution) was added. The reaction was carried out at room temperature for 2 hours. The reaction was quenched with water (20 mL), extracted with ethyl acetate (50 mL × 3), concentrated, and purified by column chromatography to give 702 mg of compound 3-R00-2, yield: 95%.

[0419] MS-ESI: m / z 578.8 [1 / 2M+H] + .

[0420] Step 6

[0421] Under a nitrogen atmosphere, compound 3-R00-2 (0.299 g, 0.259 mmol) and PyBOP (0.162 g, 0.311 mmol) were dissolved in anhydrous N,N-dimethylformamide (4 mL). A solution of the dendritic polymer 1-PEG2K (0.400 g, 5.177 μmol) and NMM (0.209 g, 2.071 mmol) in anhydrous N,N-dimethylformamide (4 mL) was added dropwise, and the reaction was carried out at 38 °C for 3 hours. The reaction solution was diluted with acetonitrile and then filtered through ultrafiltration (10 kDa). The apparatus was used to purify the crude product by ultrafiltration with acetonitrile (0.4 L, 13 V) to obtain 0.470 g of crude product. This crude product was dissolved in pure water, filtered through a 0.22 μm membrane, and lyophilized to give 0.465 g of compound 3-R00, yield: 88%.

[0422] 1 1H NMR indicates 25 drug molecules / dendritic structures. The actual molecular weight is approximately 102.0 kDa (23.76% by weight, compound 3).

[0423] 1 H NMR (400MHz, CD3OD) δ6.40-8.51(m,514H), 4.13-4.72(m,123H), 3.40-4.10(m,5900H) ),3.33-3.36(s,93H),2.79-3.25(m,73H),2.22-2.74(m,206H),0.61-1.92(m,810H).

[0424] Example 14: Preparation of compound 3-S00

[0425]

[0426] first step

[0427] Under a nitrogen atmosphere, compound 1-Z01-J01 (5 g, 18.3 mmol) was dissolved in dichloromethane (50 mL), and triethylamine (5.5 g, 54.9 mmol) was added. The mixture was cooled to 0 °C, and acetic anhydride (2 g, 18.7 mmol) was added dropwise. The reaction was allowed to proceed overnight at room temperature. The mixture was washed with water (20 mL × 2), and the aqueous phases were combined. The pH was adjusted to 1 with concentrated hydrochloric acid, and the mixture was extracted with ethyl acetate (50 mL × 3). After concentration, 4.7 g of compound 1-Z01-Q-02 was obtained, with a yield of 91%.

[0428] MS-ESI: m / z 280.2 [M+H] + .

[0429] Step 2

[0430] Under a nitrogen atmosphere, compound 1-Z01-Q-02 (4.5 g, 16.1 mmol) and trimethylsilylethanol (5.7 g, 48.3 mmol) were dissolved in dichloromethane (5 mL), cooled to 0 °C, and thionyl chloride (1.9 g, 16.1 mmol) was added. The mixture was reacted overnight at room temperature. After concentration, the solution was purified by column chromatography to give 1.6 g of compound 1-Z01-Q-03, yield: 26%.

[0431] MS-ESI: m / z 380.3 [M+H] + .

[0432] 1 H NMR(400MHz, CDCl3)δ7.39-7.28(m,5H),6.20(d,J=7.6Hz,1H),5.14(s,2H),4.61-4.63(m,1H), 4.19-4.04(m,2H),2.21-2.26(m,3H),2.02-1.94(m,4H),1.02-0.90(m,2H),0.05-0.02(m,9H).

[0433] Step 3

[0434] Compound 1-Z01-Q-03 (0.9 g, 2.3 mmol) was dissolved in tetrahydrofuran (10 mL), and wet Pd / C (180 mg, 10 wt.%) was added. The mixture was purged with hydrogen three times and reacted overnight at room temperature. After filtration and concentration, 621 mg of compound 1-Z01-Q-TMS was obtained, yield: 93%.

[0435] MS-ESI: m / z 290.3 [M+H] + .

[0436] 1 H NMR (400MHz, CDCl3) δ6.75 (d, J = 6.9Hz, 1H), 4.54-4.42 (m, 1H), 4.22-4.07 (m, 2H), 2.5 6-2.32(m,2H),2.24-2.12(m,1H),2.09-1.90(m,4H),1.03-0.90(m,2H),0.00(s,9H).

[0437] Step 4

[0438] Under a nitrogen atmosphere, compound 1-Z01-Q-TMS (400 mg, 1.3 mmol) was dissolved in dichloromethane (20 mL), and DMAP (56 mg, 0.46 mmol), 3-TMS (962 mg, 0.92 mmol), and DCC (285 mg, 1.3 mmol) were added. The mixture was reacted overnight at room temperature. Then, 150 mg of 1-Z01-Q-TMS (0.5 mmol) and DCC (143 mg, 0.6 mmol) were added, and the mixture was reacted overnight at room temperature. After filtration and concentration, the mixture was purified by column chromatography to give 560 mg of compound 3-S00-1, yield: 33%.

[0439] MS-ESI: m / z 1314.3 [M+H] + .

[0440] Step 5

[0441] Under a nitrogen atmosphere, compound 3-S00-1 (560 mg, 0.426 mmol) was dissolved in tetrahydrofuran (8 mL), and TBAF (4.3 mL, 4.3 mmol, 1.0 mol / L THF solution) was added. The reaction was allowed to proceed for 2 hours. The mixture was quenched with water (10 mL), extracted with ethyl acetate (50 mL × 3), concentrated, and purified by column chromatography to give 420 mg of compound 3-S00-2, yield: 86%.

[0442] MS-ESI: m / z 1142.3 [M+H] + .

[0443] Step 6

[0444] Under a nitrogen atmosphere, compound 3-SO0-2 (0.074 g, 0.065 mmol) and PyBOP (0.041 g, 0.078 mmol) were dissolved in anhydrous N,N-dimethylformamide (1 mL), and a solution of dendritic polymer 1-PEG2K (0.100 g, 1.30 μmol) and NMM (0.026 g, 0.258 mmol) in anhydrous N,N-dimethylformamide (1 mL) was added dropwise. The reaction was carried out at room temperature for 2 hours. The reaction solution was diluted with acetonitrile and then filtered through ultrafiltration (10 kDa). The apparatus was used to purify the compound 3-SO0 using ultrafiltration with acetonitrile (0.3 L, 25 V), and the crude product was concentrated to obtain 0.124 g. The crude product was dissolved in pure water, filtered through a 0.22 μm membrane, and lyophilized to give 0.12 g of compound 3-SO0, yield: 87%.

[0445] 1 1H NMR indicates 30 drug molecules / dendritic structures. The actual molecular weight is approximately 107.3 kDa (27.12% by weight, compound 3).

[0446] 1 H NMR (400MHz, CD3OD) δ6.40-8.52(m,609H), 4.19-4.68(m,184H), 3.40-4.10(m,5900H) ,3.33-3.36(s,102H),2.79-3.20(m,102H),2.22-2.74(m,146H),0.61-1.92(m,767H).

[0447] Example 15: Preparation of compound 3-T00

[0448]

[0449] first step

[0450] Under a nitrogen atmosphere, compound 3 (1.0 g, 1.02 mmol) was dissolved in dichloromethane (20 mL), and a solution of NMM (322 mg, 3.18 mmol) and N,N-diisopropylethylamine (89 mg, 0.69 mmol) in dichloromethane (5 mL) was added dropwise. Glutaric anhydride (141 mg, 1.23 mmol) was then added, and the reaction was carried out at room temperature for 20 hours. Glutaric anhydride (94 mg, 0.82 mmol) was then added, and the reaction was carried out at room temperature for 3 hours. After concentration, the solution was purified by column chromatography to give 1.030 g of compound 3-T00-1, yield: 92%.

[0451] MS-ESI: m / z 1085.2 [M+H] + .

[0452] Step 2

[0453] Under a nitrogen atmosphere, compound 3-T00-1 (0.352 g, 0.325 mmol) and PyBOP (0.203 g, 0.390 mmol) were dissolved in anhydrous N,N-dimethylformamide (5 mL), and a solution of dendritic polymer 1-PEG2K (0.500 g, 6.50 μmol) and NMM (0.131 g, 1.300 mmol) in anhydrous N,N-dimethylformamide (5 mL) was added dropwise. The reaction was carried out at room temperature for 2 hours. The reaction solution was diluted with acetonitrile and then filtered through ultrafiltration (10 kDa). The apparatus was used to purify the compound 3-T00 by ultrafiltration with acetonitrile (0.5 L, 25 V), and the crude product was concentrated to obtain 0.585 g. The crude product was dissolved in pure water, filtered through a 0.22 μm membrane, and lyophilized to obtain 0.580 g of compound 3-T00, yield: 84%.

[0454] 1 1H NMR indicates 31 drug molecules / dendritic structures. The actual molecular weight is approximately 106.6 kDa (28.19% by weight, compound 3).

[0455] 1 H NMR (400MHz, CD3OD) δ6.40-8.52(m,623H), 4.19-4.67(m,173H), 3.40-4.10(m,5900H) ),3.33-3.36(s,95H),2.75-3.10(m,73H),2.22-2.74(m,217H),0.61-1.92(m,613H).

[0456] Biological evaluation

[0457] Test Example 1: Pharmacokinetic Study of Different Compounds Administered by a Single Intravenous Injection to Rats

[0458] 1. Test sample

[0459] Docetaxel (G1), and the disclosed compounds 1-Z00 (G2), 1-Z00-J (G3), 1-Z00-K (G4), 1-Z00-M (G5), and 1-C00 (G6).

[0460] Sample preparation method:

[0461] (1) Preparation of docetaxel: Take commercially available docetaxel injection (0.5mL:20mg) and dilute it with physiological saline to 4mg / mL;

[0462] (2) Preparation of other test compounds: The compounds were dissolved in DMSO with a final concentration of 5% and then diluted with physiological saline to 4 mg / mL.

[0463] 2. Experimental animals

[0464] Female SD rats, weighing 160-180g, 6 weeks old, SPF grade

[0465] 3. Test methods

[0466] Blood samples were collected at different time points after intravenous administration (dose: 10 mg / kg) (G1 group: 0, 0.0833, 0.5, 1, 4, 8, 24 h; G2-6 group: 0, 0.0833, 1, 4, 8, 24, 48, 96, 168 h). The samples were anticoagulated with EDTAK2, separated, and then the esterase inhibitor DDVP was added. The samples were then frozen at -80℃, and the docetaxel level was measured (N=3).

[0467] Table 1 Grouping and Dosing Regimens

[0468]

[0469] 4. Experimental Results

[0470] As shown in Table 2 and Figure 1 As shown, after intravenous administration, the AUC of the 10 mg / kg 1-Z00-J group, 1-Z00-K group, and 1-Z00-M group was... 0-t All were significantly superior to other groups of drugs, and the plasma clearance rates of the 1-Z00-J and 1-Z00-M groups were even lower, indicating that they had a better sustained-release effect.

[0471] Table 2 PK parameters for each group

[0472]

[0473] 5. Experimental Conclusions

[0474] Compared to conventional docetaxel injection, which exhibits rapid plasma clearance and low exposure, 1-Z00-J, 1-Z00-K, and 1-Z00-M all demonstrate significant sustained-release effects and higher exposure. Compared to 1-C00 and 1-Z00, the compounds disclosed herein significantly increase drug exposure and have lower drug clearance rates.

[0475] Test Example 2: Pharmacokinetic Study of Different Compounds Administered as a Single Intravenous Injection to Beagles

[0476] 1. Test sample

[0477] The disclosed compounds are 1-B00, 1-Z00, 1-Z00-J, and docetaxel.

[0478] Sample preparation method:

[0479] 1-B00, 1-Z00, 1-Z00-J: 84 mg / vial of the test substance, added to an appropriate volume of physiological saline to obtain a drug solution with a concentration of 0.6 mg / mL, for intravenous injection.

[0480] Commercially available docetaxel in 84 mg / vial was dissolved in 5% DMSO + 30% PEG300 + 5% Tween80 + 60% deionized water to obtain a dosage form of 0.6 mg / ml for intravenous administration.

[0481] 2. Experimental animals

[0482] Beagle Level, sourced from Medicipua Reserve Animal Bank: 999M-004.

[0483] 3. Test methods

[0484] Administration method: Weigh the patient before administration and calculate the dosage based on body weight. Administer via slow intravenous injection. Collect approximately 1 mL of blood via the jugular vein or other suitable method. Collect plasma samples at different time points before and after administration to detect the level of free docetaxel in the plasma. For the docetaxel group, samples were collected before administration and at 0.083 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration; for other test drug groups, samples were collected before administration and at 0.083 h, 1 h, 4 h, 8 h, 24 h, 48 h, 72 h, 96 h, 120 h, 144 h, and 168 h after administration.

[0485] Table 3 Dosing Regimen

[0486]

[0487] 4. Test Results

[0488] The pharmacokinetic data for each group are shown in Table 4 below. Figure 2, Figure 3 As shown.

[0489] Table 4 Pharmacokinetic Data

[0490]

[0491] The experimental results show that 1-Z00-J has a higher in vivo exposure and a lower drug clearance rate compared to docetaxel and 1-Z00.

[0492] Test Example 3: Pharmacodynamic and hematologic evaluation of a subcutaneous xenograft tumor model of human lung cancer cells A549 in BALB / c nude mice.

[0493] 1. Test sample

[0494] Dorsey, 1-Z00, 1-Z00-J

[0495] The sample preparation methods are shown in Table 5 below:

[0496] Table 5 Preparation of test and reference solutions

[0497]

[0498] 2. Laboratory animals

[0499] BALB / c nude mice, female, 6-7 weeks old (mice's age at tumor cell inoculation), weighing 16.5–22.6 g, were purchased from Jiangsu GemPharmatech Co., Ltd., Animal Certificate No.: 320727210100243463. Housing conditions: SPF grade.

[0500] 3. Experimental Methods

[0501] 3.1 Cell Culture

[0502] A549 cells were cultured in F12K medium (Gibco) containing 10% fetal bovine serum (cellbank, Chinese Academy of Sciences / Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (cellbank.org.cn, SIBS)). When the cells reached the exponential growth phase, they were collected and resuspended in PBS to a suitable concentration for subcutaneous tumor inoculation in mice.

[0503] 3.2 Animal modeling and randomization

[0504] Female mice were subcutaneously injected with 5×10⁻⁶ cells on the right side. 6 A549 cells (0.1 mL / mouse). Day 0 was defined as the day of inoculation, with an average tumor volume of 142.96 mmHg. 3At day 13 post-cell inoculation, cells were randomly grouped according to tumor size as shown in Table 6 below (administration volume: 10 μL / g):

[0505] Table 6. Experimental Design of Antitumor Effects of Drugs in a Human Lung Cancer Cell A549 Subcutaneous Transplantation Tumor Model

[0506]

[0507] 3.3 Experimental observation and data collection

[0508] Post-tumor inoculation routine monitoring included monitoring tumor growth and the impact of treatment on normal animal behavior, including animal activity, feeding and drinking, weight gain or loss (measured twice weekly), and any abnormalities in the eyes, coat, or other areas. Clinical symptoms observed during the experiment were recorded in the raw data. Tumor volume calculation formula: Tumor volume (mm²) 3 )=1 / 2×(a×b 2 (Where a represents the major axis and b represents the minor axis). Data was collected using StudyDirector™ software (version 3.1.399.19, vendor: Studylog Systems, Inc., S. San Francisco, CA, USA), including measurements of the tumor's major and minor axes and animal weight. Raw data were directly imported into the software after measurement using a balance and vernier calipers; any changes to the data were recorded in the software.

[0509] 3.4 Management of Animal Weight Loss During the Experiment

[0510] When the weight loss of a single animal exceeds 15% (BWL≥15%), the corresponding single animal / group of animals shall be given a withdrawal period until the weight loss recovers to within 15% (BWL<15%).

[0511] When the weight of a single mouse decreases by more than 20%, it is euthanized in accordance with animal welfare regulations.

[0512] 3.5 Efficacy Evaluation Criteria

[0513] The relative tumor proliferation rate, T / C (%), is the percentage of tumor volume or weight in the treatment group and the control group at a certain time point. The calculation formula is: T / C% = TRTV / CRTV × 100% (TRTV: average RTV in the treatment group; CRTV: average RTV in the control group; RTV = Vt / V0, where V0 is the tumor volume of the animal at the time of grouping, and Vt is the tumor volume of the animal after treatment).

[0514] The relative tumor inhibition rate, TGI (%), is calculated as follows: TGI% = (1-T / C) × 100% (T and C are the relative tumor volume (RTV) or tumor weight (TW) of the treatment group and the control group at a specific time point, respectively).

[0515] 3.6 Experimental Endpoint

[0516] After the last administration, plasma was collected at the time points designed in Table 7 (using EDTA anticoagulant tubes (with the esterase inhibitor DDVP added during blood collection)). After the plasma collection was completed at the final time point of the mouse, the mouse was euthanized, the tumor was removed, and it was quick-frozen and preserved.

[0517] Table 7 Sampling Time Points in the Experiment

[0518]

[0519] 3.7 Statistical Analysis

[0520] All experimental results are expressed as mean tumor volume ± SEM (mean standard error). Statistical analysis between different groups was performed at the optimal treatment point (usually after the last dose). Two-way ANOVA was used to compare tumor volume between the treatment group and the control group to determine if there was a significant difference. p < 0.05 was considered statistically significant. All statistical analyses and graphs were performed in the R language environment (version 3.3.1). Unless otherwise specified, all tests were two-tailed tests, and a p-value less than 0.05 was considered statistically significant.

[0521] 4. Experimental Results

[0522] 4.1 Results of the study on the antitumor effect of the drug in a subcutaneous transplantation tumor model of human lung cancer cells A549: The average tumor volume of the vehicle control group mice was 1620.93 mm² on day 21 after the start of drug administration. 3 .

[0523] The mean tumor volume in the docetaxel 20 mg / kg group was 564.93 mm on day 21 after the start of docetaxel administration. 3 The tumor inhibition rate (TGI) was 66.24%, which was statistically significant compared with the control group (p<0.001).

[0524] The mean tumor volume in the 1-Z00 20 mg / kg group was 576.02 mmHg on day 21 after the start of administration. 3 The tumor inhibition rate (TGI) was 64.85%, which was statistically significant compared with the control group (p<0.001).

[0525] In the 1-Z00-J 10 mg / kg group, the mean tumor volume on day 21 after the start of administration was 285.12 mm.3 The tumor inhibition rate (TGI) was 83.02%, which was statistically significant compared with the control group (p<0.001).

[0526] Tumor growth in each treatment group and control group is shown in Tables 8 and 9 below. Figure 4 .

[0527] Table 8. Changes in tumor volume over treatment time in mice of different groups in a human lung cancer cell A549 subcutaneous transplantation tumor model.

[0528]

[0529] Note: Data are expressed as mean ± standard error.

[0530] Table 9. Efficacy analysis of each group in the subcutaneous transplantation tumor model of human lung cancer cells A549

[0531]

[0532] Note: 1. Data is expressed as mean ± standard error;

[0533] 2.T / C%=TRTV / CRTV×100%; TGI%=(1-T / C)×100%.

[0534] 3. P-value: The value was obtained by comparing the tumor volume of the treatment group with that of the control group using a two-way ANOVA.

[0535] 4.2 Results of pharmacodynamic and hematological studies of the drug in a subcutaneous transplantation model of human lung cancer cells A549

[0536] As shown in Table 10 below, compared with the solvent control group, the number of white blood cells, lymphocytes, and monocytes in the blood of mice injected intravenously with 20 mg / kg docetaxel decreased by 2-3 times, demonstrating the significant lymphatic system toxicity of docetaxel. Compared with the intravenous injection of 20 mg / kg docetaxel, the intravenous injection of 1-Z00-J with 10 mg / kg significantly improved the lymphatic system safety.

[0537] Table 10. Efficacy and Hematological Analysis of Drugs in a Subcutaneous Transplantation Tumor Model of Human Lung Cancer Cell A549

[0538]

[0539] Note: Data are expressed as mean ± standard deviation.

[0540] 5. Experimental Conclusions

[0541] Compared to regular docetaxel injection and 1-Z00, 1-Z00-J exhibits significant superior efficacy at half the dose. Furthermore, the molecular structure of 1-Z00-J significantly reduces the hematological toxicities of docetaxel injection, such as lymphopenia and neutropenia.

[0542] Test Example 4: Pharmacokinetic Study of Different Compounds Administered as a Single Intravenous Injection to Beagles

[0543] 1. Test sample

[0544] The disclosed compounds are: 3-L00, 3-M00, 3-N00, 3-O00, 3-R00 and 3-T00.

[0545] Sample preparation method:

[0546] Test substances 3-L00, 3-M00, 3-N00, 3-O00 and 3-R00 (80 mg / vial) were added to a final volume of 5% DMSO and 95% physiological saline, and vortexed to dissolve to obtain a 1.5 mg / ml dosage form.

[0547] 3-T00: 120 mg / vial of the test substance was added to a final volume of 5% DMSO and 95% physiological saline, and vortexed to dissolve to obtain a dosage form of 1.5 mg / ml.

[0548] 2. Experimental animals

[0549] Beagle Level, sourced from Medicipua Reserve Animal Bank: 999M-004.

[0550] 3. Test methods

[0551] Administration method: Weigh the patient before administration and calculate the dosage based on body weight. The administration regimen is shown in Table 11 below. Blood collection time points are 0.083h, 1h, 4h, 8h, 24h, 48h, 72h, and 96h after administration. Blood is collected via the jugular vein or other suitable method at specific time points after administration, with approximately 1mL collected per sample. Place the collected blood samples in EDTA-K2 anticoagulant blood collection tubes. Each group of blood samples requires special treatment with the addition of an esterase inhibitor. Add 1 / 40 of the blood collection volume of 200mM DDVP to the EDTA-K2 anticoagulant blood collection tube (if the blood collection volume is 1mL, add 25μL of 200mM DDVP). After collection, place the blood samples on ice and centrifuge to separate the plasma within 1 hour (centrifugation conditions: centrifugation force 2200g, 10 minutes, 2-8℃). Subsequently, the drug concentration in the plasma is measured, and pharmacokinetic data are calculated. Plasma samples are stored at -70℃ before testing.

[0552] Table 11 Dosing Regimen

[0553]

[0554] 4. Test Results

[0555] The pharmacokinetic (PK) results of 3-L00, 3-M00, 3-N00, 3-O00, 3-R00 and 3-T00 in beagle dogs are shown in the table below. Figure 5 As shown in Table 12.

[0556] Table 12 Pharmacokinetic Data

[0557]

[0558] Experimental results show that the exposure levels (AUC) of the disclosed dendritic polymers 3-L00, 3-M00, 3-N00, 3-O00, and 3-R00 are significantly higher than that of 3-T00, indicating a significant sustained-release effect. The disclosed dendritic polymers, through the combination of dendritic polymers with nitrogen-linked alkyl linkers on their side chains, can promote the release of small molecules compared to all-carbon chain linkers.

[0559] Test Example 5: Effects of a single intravenous injection of compounds 3, 3-M00, and 3-O00 on platelet count in beagle dogs

[0560] 1. Test sample

[0561] The disclosed compounds are: compound 3, 3-M00 and 3-O00.

[0562] Sample preparation method:

[0563] The test compound 3, in a dose of 120 mg / vial, was dissolved by vortexing with 5% DMSO, 5% Tween 80, and 90% physiological saline to obtain a dosage form of 1.5 mg / ml.

[0564] Test substances 3-M00 and 3-O00, in 80 mg / vial form, were added to a final volume of 5% DMSO and 95% physiological saline, and vortexed to dissolve and obtain a dosage form of 1.5 mg / ml.

[0565] 2. Experimental animals

[0566] Beagle Level, sourced from Medicipua Reserve Animal Bank: 999M-004.

[0567] 3. Test methods

[0568] Administration method: Weigh the patient before administration and calculate the dosage based on body weight. The administration regimen is shown in Table 13 below. Administer via slow intravenous injection. Collect approximately 1 mL of blood via the jugular vein or other suitable method. Collect whole blood before administration and 24 hours after administration for hematological testing.

[0569] Table 13 Dosage regimens for compounds 3, 3-M00, and 3-O00

[0570]

[0571] 4. Test Results

[0572] Table 14 Effects of compounds 3, 3-M00, and 3-O00 on platelet (PLT) counts in canines.

[0573]

[0574] The experimental results show that the dendritic polymers 3-M00 and 3-O00 of this disclosure can significantly reduce thrombocytopenia in the target of small molecule compound 3. In this experiment, three beagle dogs in the small molecule compound 3 treatment group were intolerant to a single intravenous injection of 3 mg / kg of compound 3, resulting in the death of two animals. However, the beagle dogs in the dendritic polymer 3-M00 and 3-O00 treatment groups did not die, further demonstrating that the dendritic polymers of this disclosure significantly improve the safety of small molecule compound 3.

[0575] Test Example 6: Efficacy of 3-L00, 3-M00, 3-N00, 3-O00, 3-R00 and 3-T00 on subcutaneous xenografts of human acute lymphoblastic leukemia RS4;11 mice

[0576] 1. Test sample

[0577] 3-L00, 3-M00, 3-N00, 3-O00, 3-R00 and 3-T00.

[0578] Sample preparation method:

[0579] 3-L00, 3-M00, 3-N00, 3-O00, 3-R00 and 3-T00: Add 0.25 mL of DMSO to each vial to dissolve the compound to obtain a working stock solution of 20 mg / mL, then add 4.75 mL of physiological saline and vortex to dissolve to obtain a dosage form of 1 mg / mL.

[0580] 2. Experimental cells and animals

[0581] Human acute lymphoblastic leukemia cells (RS4;11) were purchased from the American Type Culture Collection. RS4;11 cells were cultured in 10-cm dishes under the following conditions: RPMI 1640 medium (Gibco) with 10% fetal bovine serum (Gibco), penicillin, and streptomycin, in an incubator at 37°C with 5% CO2. Cells were passaged twice a week. When the cells reached the exponential growth phase, they were collected, counted, and seeded.

[0582] NOD-Scid mice, 5 weeks old, female, were purchased from Beijing Huafukang Biotech Co., Ltd. Production License Number: SCXK(Beijing)2019-0008; Animal Qualification Certificate Numbers: 110322211100992176, 110322211101069023. Feeding environment: SPF level.

[0583] The use and welfare of the experimental animals in this study were carried out in accordance with the regulations of the "Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC)". The health status and mortality of the animals were monitored daily. Routine examinations included observing the effects of the test substances and drugs on the daily behavior of the animals, such as behavioral activities, body weight changes, and external signs.

[0584] 3. Test methods

[0585] Each mouse was subcutaneously inoculated with 1×10 7 RS4;11 cells. When the tumors grew to 150-200 mm 3 , they were grouped according to tumor volume, and the drug was injected intravenously (IV) once a week (QW) for a total of 4 times. The injection volume was 0.1 mL / 10 g body weight; the dosing doses and dosing regimens are shown in Table 15 below. The experimental index was to investigate the effect of the drug on tumor growth, and the specific index was T / C% or tumor growth inhibition rate (TGI%).

[0586] The tumor diameter was measured twice a week with a vernier caliper. The formula for calculating the tumor volume (V) was:

[0587] V = 1 / 2 × a × b 2 where a and b represent the length and width, respectively.

[0588] T / C(%) = (T - T0) / (C - C0) × 100

[0589] where 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.

[0590] Tumor growth inhibition rate % (TGI%) = 100 - T / C(%).

[0591] Table 15 Dosing regimen

[0592]

[0593] Note: Random grouping, the first dosing time was D0; IV: Intravenous injection.

[0594] 4. Test results

[0595] Table 16 Efficacy of the compound on subcutaneous transplanted tumors in human acute lymphoblastic leukemia RS4;11 mice

[0596]

[0597]

[0598] The experimental results showed that in the subcutaneous xenograft model of human acute lymphoblastic leukemia RS4;11 mice, 3-T00 could not effectively release small molecule compound 3 and had no antitumor activity; while the linkers of 3-L00, 3-M00, 3-N00, 3-O00, and 3-R00 could effectively promote the release of small molecule compound 3 and had significant antitumor activity.

Claims

1. A macromolecule comprising: i) a dendrimer D having surface amino groups, wherein at least two different end groups are covalently attached to the surface amino groups of the dendrimer: ii) a first end group which is a pharmaceutically active agent or a residue thereof A comprising a hydroxyl, an amino or a thiol group; iii) a second end group which is a pharmacokinetic modifier; wherein said first end group is covalently attached to a surface amino group of said dendrimer via a linker -X1-L-X2-, X1is a linker to a pharmaceutically active agent, X2is a linker to the dendrimer, X1and X2are both -C(O)-, and L is C 1-10 straight or branched chain alkylene, said C 1-10 straight or branched chain alkylene is substituted with one or more -NR1R2, wherein R1, R2are each independently selected from the group consisting of hydrogen, C 1-6 alkyl and C(O)R4, R4is selected from the group consisting of C 1-6 alkyl, and R1, R2are not both hydrogen; wherein the pharmacokinetic modifier is selected from polyethylene glycol; the dendrimer D is selected from polylysine.

2. The macromolecule according to claim 1, wherein R1, R2 are each independently selected from the group consisting of hydrogen and C 1-6 alkyl, and R1, R2 are not simultaneously hydrogen.

3. The macromolecule of claim 1, wherein the C 1-10 a straight-chain or branched-chain alkylene group is a C 1-6 a straight-chain or branched-chain alkylene group.

4. The macromolecule according to claim 1, wherein in the linker -X1-L-X2-, X1 is -C(O)-, attached to the pharmaceutically active agent or residue thereof A; X2 is -C(O)-, attached to the surface amino group of the dendrimer D to form an amide bond; the macromolecule structure is as follows:

5. The macromolecule according to claim 1, wherein the pharmaceutically active agent is selected from anesthetics, antacids, antibodies, anti-infectives, biologicals, cardiovascular drugs, contrast media, diuretics, blood supplements, immunosuppressants, hormones, nutraceuticals, ophthalmic drugs, pain management agents, respiratory drugs, adjuvants, anti-arthritic drugs, anti-convulsants, anti-histamines, anti-inflammatory drugs, anti-ulcer drugs, behavior modification drugs, cancer drugs, central nervous system drugs, contraceptives, diabetes management drugs, fertility drugs, growth promoters, hemostatic drugs, immunostimulants, muscle relaxants, obesity management drugs, osteoporosis drugs, peptides, sedatives and tranquilizers, urinary tract acidifiers, or vitamins.

6. The macromolecule according to claim 5, wherein the pharmaceutically active agent is selected from cancer drugs.

7. The macromolecule according to claim 1, wherein the pharmaceutically active agent is selected from docetaxel, irinotecan, gemcitabine, capecitabine, decitabine, azacitidine, doxorubicin, epirubicin, trabectedin, rubitecan, bortezomib, eribulin, selinexor, venetoclax, tesidolumab, pemetrexed, cabazitaxel, cabozantinib, onartamab, the following compounds 2 and 3, 8. The macromolecule according to claim 7, wherein the pharmaceutically active agent is docetaxel, compound 2, compound 3.

9. The macromolecule according to claim 1, wherein the polyethylene glycol has a molecular weight in the range of 220 to 5500 Da.

10. The macromolecule according to claim 9, wherein the polyethylene glycol has a molecular weight in the range of 1000-5500 Da.

11. The macromolecule according to claim 1, wherein the dendritic polymer D is selected from BHALys[Lys]. 16 BHALys[Lys] 32 Or BHALys[Lys] 64 .

12. A pharmaceutical composition comprising the macromolecule according to any one of claims 1 to 11 and a pharmaceutically acceptable carrier.

13. Use of the macromolecule according to any one of claims 6 to 8 in the manufacture of a medicament for the treatment of a tumor.

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