Drug conjugates for the treatment of eye disorders

Abstract

The present invention relates to drug conjugates comprising hyaluronic acid (HA) hydrogel microspheres or pharmaceutically acceptable salts thereof, pharmaceutical compositions and methods of using such conjugates for the treatment of ocular disorders, and methods of making the conjugates.

Description

[Technical Field]

[0001] The present invention relates to drug conjugates comprising hyaluronic acid (HA) hydrogel microspheres or pharmaceutically acceptable salts thereof, pharmaceutical compositions and methods of using such conjugates for the treatment of ocular disorders, and methods of making the conjugates. [Background technology]

[0002] A major cause of blindness is the inadequate treatment of certain ocular diseases. A major limitation is the lack of suitable options for introducing drugs or therapeutic agents into the eye and maintaining them in the eye at therapeutically effective concentrations for the required period of time. Systemic administration is not an ideal solution because, in many cases, unacceptably high levels of systemic administration are required to achieve effective intraocular concentrations, accompanied by an increased incidence of unacceptable side effects. Simple eye drops or ocular application are often not an acceptable alternative because the drug may be rapidly washed out by lacrimal action or decline from the eye into the systemic circulation. Topical eye drop therapy is limited by poor absorption, the need for frequent and / or chronic administration over a period of days to years, rapid aqueous humor turnover, and the production and movement of tear film and other contributors that may effectively remove the therapeutic agent long before therapy is completed or an adequate dose has been delivered.

[0003] Intraocular injection has the advantage of providing improved bioavailability to the target location (e.g., the retina) in the eye compared to other delivery mechanisms, such as topical delivery. However, intraocular injection also has drawbacks and can cause a variety of different complications. For example, intravitreal injection can result in the delivery of undesirably high concentrations of the therapeutic agent to the target location or other locations, especially if the therapeutic agent is relatively soluble. Furthermore, intraocular injection is very uncomfortable for the patient. Furthermore, because intraocular injection itself can cause complications such as endophthalmitis and retinal detachment, it is highly desirable to have as long a period between injections as possible while maintaining therapeutic levels of the drug in the eye.

[0004] In addition to the above, the therapeutic agent delivered by intravitreal injection can often be dispersed quickly in the eye after injection, so that the duration of action can be insufficient.This insufficient duration can require more frequent injection, which is particularly undesirable.For example, Ranibizumab and Pegaptanib are administered to patients by intraocular injection every 4 weeks and 6 weeks, respectively, which is a very unpleasant experience for patients. Summary of the Invention [Problem to be solved by the invention]

[0005] Thus, there is growing recognition that the field of ophthalmology would benefit from longer-lasting formulations that would benefit patient care and ocular health by providing long-term delivery of therapeutic agents to the eye while minimizing problems associated with patient compliance with prescribed therapeutic medical regimens.

[0006] Expression of vascular endothelial growth factor (VEGF), a signaling protein produced by cells that stimulates vasculogenesis and angiogenesis, plays an important role in various ocular conditions, including macular degeneration and certain forms of retinopathies.

[0007] Various medications are on the market to treat such eye conditions, including ranibizumab, aflibercept, and pegaptanib, which are administered to patients via intraocular injection every four and eight weeks.

[0008] In view of the above, there is a need to provide a dosage form that at least partially overcomes these drawbacks. [Means for solving the problem]

[0009] For this purpose, a drug conjugate or a pharmaceutically acceptable salt thereof comprising a hyaluronic acid (HA) hydrogel microsphere comprising crosslinked HA chains to which a plurality of drug moieties are covalently and reversibly conjugated, or a pharmaceutically acceptable salt thereof, wherein said drug conjugate comprises each of the following units:

[0010] [ka] [In the formula, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each R a1 -H, C 1~10 independently selected from the group consisting of alkyl, ammonium ion, tetrabutylammonium ion, cetyltrimethylammonium ion, alkali metal ion, and alkaline earth metal ion; Each-R a2 are independently -H or C 1~10 is alkyl, each -X- and -Y- is independently a carbonyl group or is absent; each -X'-, -Y'- is independently a spacer moiety or absent; Each -D can independently 1 a VEGF-neutralizing drug moiety covalently and reversibly conjugated to Each-L 1- is independently a reversible linker moiety; Each-L 2 - is independently a spacer moiety or absent; Each-L 3 -, -L 4 -, -L 5 - is independently a linking moiety or absent; each -BA is independently a blocking agent] or a pharmaceutically acceptable salt thereof, [Brief explanation of the drawings]

[0011] [Figure 1] FIG. 1 shows a schematic of an embodiment of a mechanism for precipitating and isolating a polymer. [Figure 2] FIG. 2 shows a schematic of another embodiment of a mechanism for precipitating and isolating a polymer. [Figure 3] FIG. 3 shows a schematic representation of yet another embodiment of a mechanism for precipitating and isolating a polymer. DETAILED DESCRIPTION OF THE INVENTION

[0012] Within the meaning of the present invention, the terms are used as follows:

[0013] As used herein, the term "about" in conjunction with a numerical value is used to indicate a range of the numerical value plus or minus 20% of the numerical value, in certain embodiments, 15% of the numerical value, in certain embodiments, 10% of the numerical value, and in certain embodiments, 5% of the numerical value, inclusive. For example, the phrase "about 200" is used to mean a range of 200 + / - 20% (inclusive), i.e., 160 to 240 (inclusive), in certain embodiments, 200 + / - 15% (inclusive), i.e., 170 to 230 (inclusive), in certain embodiments, 200 + / - 10% (inclusive), i.e., 180 to 220 (inclusive), and in certain embodiments, 200 + / - 5% (inclusive), i.e., 190 to 210 (inclusive).

[0014] As used herein, the term "polysaccharide," also known as "glycan," refers to a compound consisting of glycosidically linked monosaccharide moieties. Typically, the term is used for compounds consisting of multiple glycosidically linked monosaccharide moieties, e.g., more than 10 monosaccharide moieties.

[0015] As used herein, the term "functionalized hyaluronic acid" refers to any hyaluronic acid derivative that can result from chemical or enzymatic functionalization or modification of natural hyaluronic acid. In particular, the term refers to any hyaluronic acid derivative that can result from chemical modification or functionalization at the carboxylic acid group.

[0016] As used herein, the term "microsphere" refers to a micron-scale particle that is typically composed of a solid or semi-solid material and is substantially spherical. Typically, the average diameter of the microspheres of the present invention ranges from about 1 μm to about 1000 μm, such as from about 10 μm to about 500 μm, or from about 50 μm to about 500 μm, as determined by microscopy, such as flow microscopy, or laser diffraction, or any other suitable method.

[0017] As used herein, the term "dispersed phase" refers to a phase containing particles or droplets of any size and nature that are distributed throughout or dispersed within a continuous phase. The diameter of the droplets within the dispersed phase can range from about 1 μm to about 5000 μm, such as from about 10 μm to about 1000 μm, or such as from about 50 μm to about 500 μm. In the dispersed phase found in emulsions of the present invention, the average diameter of the droplets in the dispersed phase is typically in the range of 1 μm to about 1000 μm, such as from 10 μm to about 500 μm, or such as from about 50 μm to about 500 μm.

[0018] As used herein, the term "continuous phase" or "continuous phase solution" refers to a fluid phase in which solid or fluid particles or droplets are distributed.

[0019] As used herein, the term "emulsion" refers to a fluid system in which droplets of one liquid are dispersed in another liquid in which it is not soluble or miscible. An emulsion is called an oil / water (o / w) emulsion when the dispersed phase is organic and the continuous phase is water or an aqueous solution, or a water / oil (w / o) emulsion when the dispersed phase is water or an aqueous solution and the continuous phase is an organic liquid.

[0020] As used herein, the term "suspension polymerization" refers to a polymerization process in which a polymer, such as a hydrogel, is formed in monomer or monomer-solvent droplets in a continuous phase that is insoluble in both the monomer and the polymer being formed. As the monomer is converted to polymer, the droplets transform into sticky, viscous monomer and / or polymer particles, which gradually become spherical, solid polymer particles or microspheres. In the context of the present invention, it is understood that the aforementioned monomers correspond to the first and second functionalized HA, and the polymer that is formed corresponds to the HA hydrogel.

[0021] As used herein, the term "drug" refers to a substance used in the treatment, cure, prevention, or diagnosis of disease, or otherwise used to improve the physical or mental well-being of a patient. When a drug is conjugated to another moiety, the portion of the resulting product that is derived from the drug is called the "drug moiety."

[0022] As used herein, the term "VEGF-neutralizing drug (moiety)" refers to a drug (moiety) that exerts its pharmaceutical effect by neutralizing the action of vascular endothelial growth factor (VEGF). The action of VEGF can be neutralized by a drug that binds to a VEGF receptor or by a drug that binds to VEGF itself, thus blocking or reducing the effective binding of VEGF to its receptor. Alternatively, the neutralizing effect can be achieved by inhibiting or interfering with the expression and production of VEGF or by interfering with VEGF signaling.

[0023] As used herein, the term "anti-VEGF antibody" refers to an antibody capable of binding to VEGF with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent or drug in targeting VEGF. In one embodiment, the extent of binding of an anti-VEGF antibody to an unrelated, non-VEGF protein is less than about 10% of the binding of the antibody to VEGF, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, an antibody that binds to VEGF has an affinity of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., ≦10 -8 M or less, e.g. 10 -8 M~10 -13 M or for example 10 -9 M~10 -13 In certain embodiments, the anti-VEGF antibody binds to an epitope of VEGF that is conserved among VEGFs from different species.

[0024] As used herein, the term "antibody" is used in the broadest sense and encompasses a variety of antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired antigen-binding activity.

[0025] As used herein, the term "angiogenesis" refers to the process by which new blood vessels form from pre-existing vessels. Disorders associated with pathological angiogenesis can be treated by the drug conjugates, pharmaceutical compositions, and methods of the invention. These diseases include both non-neoplastic disorders and cell proliferation disorders. Non-neoplastic disorders include, but are not limited to, ocular conditions (e.g., retinopathies, such as proliferative diabetic retinopathy, choroidal neovascularization (CNV), age-related macular degeneration (AMD), diabetic and other ischemia-related retinopathies, diabetic macular edema (DME), pathological myopia, von Hippel-Lindau disease, ocular histoplasmosis, retinal vein occlusions (central (CRVO) and branch (BRV)). O) morphology), corneal neovascularization, retinal neovascularization, retinopathy of prematurity (ROP), familial exudative vitreoretinopathy (FEVR), Coats' disease, Norrie's disease, osteoporosis-pseudoglioma syndrome (OPPG), subconjunctival hemorrhage, rubeosis, ocular neovascular diseases, neovascular glaucoma and hypertensive retinopathy, autoimmune diseases, unwanted or abnormal hypertrophy, arthritis, psoriatic arthritis, psoriatic plaques, sarcoidosis, atherosclerosis, Atherosclerosis, atherosclerotic plaques, arteriosclerosis, vascular restenosis, arteriovenous malformations, meningiomas, hemangiomas, angiofibromas, thyroid hyperplasia, corneal and other tissue transplants, pulmonary inflammation, acute lung injury, sepsis, primary pulmonary hypertension, malignant pulmonary effusion, cerebral edema, synovial inflammation, pannus formation in RA, myositis ossificans, hypertrophic bone formation, osteoarthritis, refractory ascites, polycystic ovarian disease, endometriosis, third-space fluid disorders, chronic asthma, uterine fibroids, preterm labor, chronic inflammation such as IBD, inflammatory kidney disease, diseases occurring after transplantation, renal allograft rejection, nephrotic syndrome, unwanted or abnormal tissue mass growth, hemophilic joints, hypertrophic scars, inhibition of hair growth, Osler-Weber syndrome, pyogenic granulomas, retrolental fibroplasia, scleroderma, trachoma, vascular adhesions, synovitis, dermatitis, preeclampsia, ascites, pericardial effusion, and pleural effusion.

[0026] As used herein, the term "ocular disorder" includes any ocular disorder or condition associated with pathological angiogenesis. Ocular disorders may be characterized by altered or uncontrolled growth and / or invasion of new blood vessels into structures of ocular tissues, such as the retina or cornea.

[0027] As used herein, the term "administered by injection" or "injectability" refers to a combination of factors such as a certain force applied to the plunger of a syringe containing a drug conjugate described herein that is swellable in a liquid at a certain concentration (w / v) and a certain temperature, a needle of a given bore diameter connected to the outlet of such a syringe, and the time required to expel a certain volume of drug conjugate from the syringe through the needle.

[0028] As used herein, the term "primary or secondary amine-containing moiety of drug DH" refers to a portion of a drug that includes at least one primary or secondary amine functional group, which drug may optionally have one or more further functional groups including one or more additional primary and / or secondary amine functional groups.

[0029] As used herein, the term "moiety" refers to a portion of a molecule that lacks one or more atoms compared to the corresponding reagent. For example, when a reagent of the formula "HXH" reacts with another reagent and becomes part of a reaction product, the corresponding portion of the reaction product has the structure "HX-" or "-X-", where each "-" indicates a bond to another moiety. Thus, the drug moiety is released as a drug from the reversible linkage.

[0030] When an arrangement or chemical structure of atoms is provided that connects two moieties or interrupts one moiety, it is understood that the arrangement or chemical structure may be connected to the two moieties in either orientation, unless expressly stated otherwise. For example, the moiety "-C(O)N(R x )-" is "-C(O)N(R x )-" or "-N(R x It may be linked to two moieties as "C(O)-" or may interrupt one moiety.

[0031] As used herein, the term "protecting group moiety" refers to a moiety that reversibly connects to a functional group, preventing the functional group from reacting with, for example, another functional group. Suitable alcohol (-OH) protecting groups include, for example, acetyl, benzoyl, benzyl, β-methoxyethoxymethyl ether, dimethoxytrityl, methoxymethyl ether, methoxytrityl, p-methoxybenzyl ether, methylthiomethyl ether, pivaloyl, tetrahydropyranyl, trityl, trimethylsilyl, tert-butyldimethylsilyl, tri-isopropylsilyloxymethyl, triisopropylsilyl ether, methyl ether, and ethoxyethyl ether. Suitable carbonyl protecting groups include, for example, acetals and ketals, acylals, and dithianes. Suitable carboxylic acid protecting groups include, for example, methyl esters, benzyl esters, tert-butyl esters, 2,6-dimethylphenol, 2,6-diisopropylphenol, 2,6-di-tert-butylphenol, silyl esters, orthoesters, and oxazolines. Suitable phosphate protecting groups are, for example, 2-cyanoethyl and methyl.

[0032] As used herein, the term "amine protecting group moiety" refers to a moiety used for reversible protection of an amine functionality during a chemical reaction process, preventing the amine from reacting, for example, with another functional group.

[0033] As used herein, the term "reducing agent" refers to a chemical compound or element that loses or donates electrons to an electron acceptor, such as an oxidizing agent, in a redox chemical reaction.

[0034] As used herein, the term "oxidizing agent" refers to a chemical compound that is capable of oxidizing other chemical compounds.

[0035] As used herein, the term "reagent" means a chemical compound that contains at least one functional group for reaction with a functional group of another chemical compound or drug. It is understood that a drug that contains a functional group is also a reagent.

[0036] It will be appreciated by those skilled in the art that the drug conjugates of the present invention, or pharmaceutically acceptable salts thereof, are prodrugs. As used herein, the term "prodrug" refers to a compound represented by the formula -L 1 -L 2 A prodrug refers to a drug moiety that is reversibly and covalently conjugated to hyaluronic acid via a -moiety. A prodrug refers to a drug moiety that is reversibly and covalently attached to hyaluronic acid via a -D or -D moiety. + in the form of its corresponding drug DH or D. In other words, the prodrug contains at least one -L 1 -L 2 -moiety. Such prodrugs or conjugates release the previously conjugated drug moiety in the form of the free or unmodified drug.

[0037] As used herein, the term "reversible linkage" or "biodegradable linkage" refers to a linkage that is cleavable under physiological conditions, i.e., aqueous buffer at pH 7.4 and 37°C, in the absence of enzymes, with a half-life ranging from 1 hour to 6 months, e.g., 10 hours to 4 months, e.g., 1 day to 3 months, 2 days to 2 months, or 3 days to 1 month. However, it is understood that a reversible linkage may also be cleavable under other conditions, e.g., at a different pH or a different temperature, with the test to determine reversibility being performed under the above physiological conditions (aqueous buffer, pH 7.4, 37°C). Thus, a "stable linkage" is a linkage that has a half-life under physiological conditions of greater than 6 months.

[0038] As used herein, the term "C 1~4 "Alkyl," alone or in combination, means a straight or branched chain alkyl moiety having 1 to 4 carbon atoms. When present at the terminal end of a molecule, it is 1~4 Examples of alkyl are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. 1~4 When linked by an alkyl, such C 1~4Examples of alkyl groups are -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -CH(C2H5)-, and -C(CH3)2-. 1~4 Each hydrogen of an alkyl carbon may be optionally replaced by a substituent as defined below. 1~4 The alkyl may be interrupted by one or more moieties as defined below.

[0039] As used herein, the term "C 1~6 "Alkyl," alone or in combination, means a straight-chain or branched alkyl moiety having 1 to 6 carbon atoms. When present at the terminal end of a molecule, straight-chain and branched C 1~6 Examples of alkyl groups are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethylpropyl. 1~6 When linked by an alkyl group, such C 1~6 Examples of alkyl groups are -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -CH(C2H5)- and -C(CH3)2-. 1~6 Each hydrogen atom of the carbon may be optionally replaced by a substituent as defined below. 1~6 The alkyl may be interrupted by one or more moieties as defined below.

[0040] Therefore, "C 1~10 Alkyl," "C 1~20 Alkyl," "C 8~24 Alkyl" or "C 1~50 "Alkyl" means an alkyl chain having 1 to 10, 1 to 20, 8 to 24, or 1 to 50 carbon atoms, respectively; C 1~10 , C 1~20 , C 8~24 or C 1~50Each hydrogen atom of the carbon may be optionally replaced by a substituent as defined below. 1~10 Alkyl, C 1~20 Alkyl, C 8~24 Alkyl or C 1~50 The alkyl may be interrupted by one or more moieties as defined below.

[0041] As used herein, the term "C 2~6 "Alkenyl," alone or in combination, means a straight or branched hydrocarbon moiety containing at least one carbon-carbon double bond, having 2 to 6 carbon atoms. When present at the end of the molecule, examples are -CH=CH2, -CH=CH-CH3, -CH2-CH=CH2, -CH=CHCH2-CH3, and -CH=CH-CH=CH2. When two portions of the molecule are C 2~6 When linked by an alkenyl group, such C 2~6 An example of an alkenyl is -CH=CH-. 2~6 Each hydrogen atom of the alkenyl moiety may be optionally replaced by a substituent as defined below. 2~6 The alkenyl may be interrupted by one or more moieties as defined below.

[0042] Hence the term "C 2~10 alkenyl," "C 2~20 alkenyl" or "C 2~50 The term "alkenyl," alone or in combination, means a straight-chain or branched hydrocarbon moiety containing at least one carbon-carbon double bond, having 2 to 10, 2 to 20, or 2 to 50 carbon atoms, respectively. 2~10 Alkenyl, C 2~20 Alkenyl or C 2~50 Each hydrogen atom of the alkenyl group may be optionally replaced by a substituent as defined below. 2~10 Alkenyl, C 2~20 Alkenyl or C 2~50 The alkenyl may be interrupted by one or more moieties as defined below.

[0043] As used herein, the term "C 2~6 "Alkynyl," alone or in combination, means a straight-chain or branched hydrocarbon moiety containing at least one carbon-carbon triple bond, having 2 to 6 carbon atoms. When present at the end of the molecule, examples are -C≡CH, -CH2-C≡CH, -CH2-CH2-C≡CH, and -CH2-C≡C-CH3. When two parts of the molecule are connected by an alkynyl group, an example is -C≡C-. C 2~6 Each hydrogen atom of the alkynyl group may be optionally replaced by a substituent as defined below. Optionally, one or more double bonds may be present. Optionally, C 2~6 The alkynyl may be interrupted by one or more moieties as defined below.

[0044] Therefore, as used herein, the term "C 2~10 alkynyl", "C 2~20 alkynyl" and "C 2~50 The term "alkynyl," alone or in combination, means a straight-chain or branched hydrocarbon moiety containing at least one carbon-carbon triple bond, having 2 to 10, 2 to 20, or 2 to 50 carbon atoms, respectively. 2~10 Alkynyl, C 2~20 Alkynyl or C 2~50 Each hydrogen atom of the alkynyl group may be optionally replaced by a substituent as defined below. Optionally, one or more double bonds may be present. Optionally, C 2~10 Alkynyl, C 2~20 Alkynyl or C 2~50 The alkynyl may be interrupted by one or more moieties as defined below.

[0045] As mentioned above, C 1~4 Alkyl, C 1~6 Alkyl, C 1~10 Alkyl, C 1~20 Alkyl, C 1~50 Alkyl, C 8~24 Alkyl, C 2~6 Alkenyl, C2~10 Alkenyl, C 2~20 Alkenyl, C 2~50 Alkenyl, C 2~6 Alkynyl, C 2~10 Alkynyl, C 2~20 Alkenyl or C 2~50 The alkynyl may be optionally interrupted by one or more moieties, which, in certain embodiments, are

[0046] [ka] [In the formula, The dashed line indicates the bond to the remainder of the moiety or reagent; -R and -R a are independently selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethylpropyl. and wherein these moieties and linkages are optionally further substituted.

[0047] As used herein, the term "C 3~10 "Cycloalkyl" means a cyclic alkyl chain having 3 to 10 carbon atoms, which may be saturated or unsaturated, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl. 3~10 Each hydrogen atom of a cycloalkyl carbon may be replaced by a substituent as defined below. 3~10 "Cycloalkyl" also includes bridged bicycles such as norbornane or norbornene.

[0048] As used herein, the term "8- to 30-membered carbopolycyclyl" or "8- to 30-membered carbopolycycle" refers to a cyclic moiety (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic rings) having 8 to 30 ring atoms, where two adjacent rings share at least one ring atom, and which may contain up to a maximum number of double bonds. In certain embodiments, an 8- to 30-membered carbopolycyclyl refers to a cyclic moiety with 2, 3, 4, or 5 rings. In certain embodiments, an 8- to 30-membered carbopolycyclyl refers to a cyclic moiety with 2, 3, or 4 rings.

[0049] As used herein, the term "3- to 10-membered heterocyclyl" or "3- to 10-membered heterocycle" refers to a ring (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic ring) having 3, 4, 5, 6, 7, 8, 9, or 10 ring atoms and which may contain up to a maximum number of double bonds, in which at least one ring atom, and up to four ring atoms, is replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and the ring is linked to the remainder of the molecule by a carbon or nitrogen atom. Examples of 3- to 10-membered heterocycles include, but are not limited to, aziridine, oxirane, thiirane, azirine, oxirene, thiylene, azetidine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazole, isothiazolidine, thiadiazole, thiadiazoline, tetrahydrofuran, tetrahydrothiophene, pyrrolidine, imidazolidine, pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, thiadiazolidine, sulfolane, pyran, dihydropyran, tetrahydropyran, imidazolidine, pyridine, pyridazine, pyrazine, pyrimidine, piperazine, piperidine, morpholine, tetrazole, triazole, triazolidine, tetrazolidine, diazepane, azepine, and homopiperazine. Each hydrogen atom of the 3- to 10-membered heterocyclyl or 3- to 10-membered heterocyclic group may be replaced by a substituent as defined below.

[0050] As used herein, the term "8- to 11-membered heterobicyclyl" or "8- to 11-membered heterobicycle" means a bicyclic heterocyclic moiety (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic ring) having 8 to 11 ring atoms and which may contain up to a maximum number of double bonds, in which at least one ring atom is shared by both rings, and at least one ring atom, up to a maximum of six ring atoms, is replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and the rings are linked to the remainder of the molecule by a carbon or nitrogen atom. Examples of 8- to 11-membered heterobicycles are indole, indoline, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzisothiazole, benzimidazole, benzimidazoline, quinoline, quinazoline, dihydroquinazoline, quinoline, dihydroquinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, decahydroisoquinoline, tetrahydroisoquinoline, dihydroisoquinoline, benzazepine, purine, and pteridine. The term 8- to 11-membered heterobicycle also includes two-ring spiro structures such as 1,4-dioxa-8-azaspiro[4.5]decane, or bridged heterocycles such as 8-aza-bicyclo[3.2.1]octane. Each hydrogen atom of an 8- to 11-membered heterobicyclyl or an 8- to 11-membered heterobicycle carbon may be replaced by a substituent as defined below.

[0051] Similarly, the term "8- to 30-membered heteropolycyclyl" or "8- to 30-membered heteropolycycle" refers to a heterocyclic moiety (fully saturated, partially saturated, or unsaturated, aromatic or non-aromatic rings) of more than two rings, in certain embodiments 3, 4, or 5 rings, having 8 to 30 ring atoms and which may contain up to a maximum number of double bonds, wherein two adjacent rings share at least one ring atom, at least one ring atom, and up to 10 ring atoms, is replaced by a heteroatom selected from the group of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and the ring is linked to the remainder of the molecule by a carbon or nitrogen atom.

[0052] structure:

[0053] [ka] Regarding the part, "Pair-R x / -R y together with the atoms to which they are attached, C 3~10 The phrase "forms a cycloalkyl, 3- to 10-membered heterocyclyl, or 8- to 11-membered heterobicyclyl" is intended to mean a -R x and -R y is understood to mean forming the following structure:

[0054] [ka] [Wherein R is C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, or 8- to 11-membered heterobicyclyl.

[0055] structure:

[0056] [ka] Regarding the part, "Pair-R x / -R ytogether with the atoms to which they are attached form a ring -A-" is an -R x and -R y is also understood to mean forming the following structure:

[0057] [ka]

[0058] As used herein, the term "π electron pair donating heteroaromatic N-containing moiety" refers to a heteroaromatic N-containing moiety that is an N-containing moiety, such as -D and -L. 1 - refers to a moiety that yields a drug DH after cleavage of the linkage between the drug moiety -D and, similarly, the corresponding DH, contains at least one, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, heteroaromatic nitrogen atom that donates a π-electron pair to the aromatic π-system. Examples of chemical structures containing such heteroaromatic nitrogen atoms that donate a π-electron pair to the aromatic π-system include, but are not limited to, pyrrole, pyrazole, imidazole, isoindazole, indole, indazole, purine, tetrazole, triazole, and carbazole. For example, in the imidazole ring below, the heteroaromatic nitrogen that donates a π-electron pair to the aromatic π-system is marked with a "#":

[0059] [ka]

[0060] The π-electron pair-donating heteroaromatic nitrogen atom does not include a heteroaromatic nitrogen atom that donates only one electron (i.e., not a pair of π electrons) to the aromatic π system, such as the nitrogen marked with "§" in the imidazole ring structure described above. The drug DH may exist in one or more tautomeric forms, for example, in which one hydrogen atom migrates between at least two heteroaromatic nitrogen atoms. In all such cases, the linker moiety is covalently and reversibly bound at the heteroaromatic nitrogen that donates the π-electron pair to the aromatic π system.

[0061] As used herein, the term "excipient" refers to a diluent, adjuvant, or vehicle with which a therapeutic agent, eg, a drug conjugate or pharmaceutical composition, is administered.

[0062] As used herein, the term "free from" a drug refers to the drug in its unmodified, pharmacologically active form, for example, after release from a conjugate.

[0063] As used herein, the term "functional group" means a group of atoms that can react with another group of atoms. Exemplary functional groups are carboxylic acid, primary amine, secondary amine, tertiary amine, maleimide, thiol, sulfonic acid, carbonate, carbamate, hydroxyl, aldehyde, ketone, hydrazine, isocyanate, isothiocyanate, phosphoric acid, phosphonic acid, haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine, disulfide, sulfonamide, sulfate, vinyl sulfone, vinyl ketone, diazoalkane, oxirane, and aziridine.

[0064] As used herein, the term "halogen" means fluoro, chloro, bromo, or iodo. In certain embodiments, halogen is fluoro or chloro.

[0065] As used herein, the term "interrupted" means that a moiety is inserted between two carbon atoms, or, if the insertion is at one of the ends of the moiety, between a carbon or heteroatom and a hydrogen atom, and in certain embodiments, between a carbon and a hydrogen atom.

[0066] As used herein, the term "pharmaceutically acceptable" means a substance that does not cause harm when administered to a patient, and preferably means approved by a regulatory body, such as the EMA (Europe) and / or the FDA (USA) and / or any other national regulatory body, for use in animals, preferably for use in humans.

[0067] As used herein, the term "pharmaceutically acceptable salt thereof" refers to a salt that retains the biological effectiveness or properties of a compound, typically without being biologically or otherwise undesirable. In certain embodiments, the compound is capable of forming acid and / or base salts due to the presence of amino and / or carboxyl functional groups or groups similar thereto. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids, such as acetate, aspartate, benzoate, besylate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, camphorsulfonate, chloride / hydrochloride, chlortheophylline, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucuronate, hippurate, hydroiodide / iodide, isethionate, lactate, lactobionate, etc. Examples of suitable salts include acetic acid salts, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methylsulfate, naphthoate, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, polygalacturonate, propionate, stearate, succinate, subsalicylate, tartrate, tosylate, and trifluoroacetate. Inorganic acids from which salts can be derived include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Organic acids from which salts can be derived include acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, and sulfosalicylic acid. Pharmaceutically acceptable base addition salts can be formed using inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from Groups I to XII of the periodic table. In certain embodiments, salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium, potassium, sodium, calcium, and magnesium salts.Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines such as naturally occurring substituted amines, cyclic amines, basic ion exchange resins, etc. Certain organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine, or tromethamine. Pharmaceutically acceptable salts can be synthesized from a basic or acidic moiety in the parent compound by conventional chemical methods.

[0068] As used herein, the term "peptide," as used herein, refers to a chain of at least two and up to and including 50 amino acid monomer moieties (which may also be referred to as "amino acid residues") linked by peptide (amide) linkages. The amino acid monomers may be selected from the group consisting of proteinogenic and non-proteinogenic amino acids and may be D- or L-amino acids. The term "peptide" also includes peptidomimetics, such as peptoids, beta-peptides, cyclic peptides, and depsipeptides, and encompasses such peptidomimetic chains having up to and including 50 monomer moieties.

[0069] As used herein, the term "protein" refers to a chain of more than 50 amino acid monomer moieties (which may also be referred to as "amino acid residues") linked by peptide linkages, preferably 12,000 or fewer amino acid monomers, e.g., 10,000 or fewer amino acid monomer moieties, 8,000 or fewer amino acid monomer moieties, 5,000 or fewer amino acid monomer moieties, or 2,000 or fewer amino acid monomer moieties linked by peptide linkages.

[0070] As used herein, the term "small molecule drug" refers to a drug that is an organic compound having a molecular weight of less than 1000 Da, for example, less than 900 Da or less than 800 Da. It is understood that nucleobase-based drug moieties, such as adenine or guanine analogs, can also be a type of small molecule drug.

[0071] As used herein, the term "medium-molecule drug" refers to a drug that is an organic compound that is not a peptide, not a protein, and has a molecular weight in the range of 1 kDa to 7.5 kDa (inclusive).

[0072] As used herein, the term "polymer" refers to a molecule comprising repeating structural units, i.e., monomers, connected by chemical bonds in a linear, cyclic, branched, cross-linked, or dendrimeric fashion, or a combination thereof, and may be of synthetic or biological origin, or a combination of both. The monomers may be the same (in which case the polymer is a homopolymer) or different (in which case the polymer is a heteropolymer). Heteropolymers may also be referred to as "copolymers," and include, for example, alternating copolymers in which different types of monomers alternate, periodic copolymers in which different types of monomers are arranged in a repeating sequence, statistical copolymers in which different types of monomers are arranged randomly, block copolymers in which different homopolymeric blocks consisting of only one type of monomer are covalently linked, and gradient copolymers in which the composition of different monomers gradually changes along the polymer chain. It is understood that a polymer may also contain one or more other moieties, such as one or more functional groups. Similarly, it is understood that a peptide or protein is a polymer even though the side chains of individual amino acid residues may differ. It is understood that covalently crosslinked polymers, such as hydrogels, may not provide a meaningful molecular weight range.

[0073] As used herein, the term "hydrogel" refers to a hydrophilic or amphiphilic polymer network composed of homopolymers or copolymers, which is insoluble due to the presence of hydrophobic interactions, hydrogen bonds, ionic interactions, and / or covalent chemical crosslinks. The crosslinks provide the network structure and physical integrity.

[0074] As used herein, the term "weight average molecular weight" or "M w " refers to the statistical average molecular weight of all molecules, expressed in units of g / mol, taking into account the weight of each molecule in determining its contribution to the molecular weight average. The higher the molecular weight of a given molecule, the more likely it is to have an M w The weight average molecular weight can be calculated by techniques known in the art that are sensitive to molecular size, such as static light scattering, small angle neutron scattering, X-ray scattering, and sedimentation velocity.

[0075] As used herein, the term "spacer" or "spacer moiety" refers to a moiety suitable for connecting two moieties.

[0076] As used herein, the term "substituted" means that one or more -H atoms of a molecule or moiety have been replaced with a different atom or group of atoms, called "substituents."

[0077] As used herein, the term "substituent" includes, in certain embodiments, halogen, -CN, -C(O)OR x1 , -OR x1 , -C(O)R x1 , -C(O)N(R x1 )(R x1a ), -S(O)2N(R x1 )(R x1a ), -S(O)N(R x1 )(R x1a ), -S(O)R x1 , -S(O)R x1 , -N(R x1 )S(O)2N(R x1a )(R x1b ), -SR x1 , -N(R x1 )(R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R x1a , -N(R x1)S(O)R x1a , -N(R x1 )C(O)OR x1a , -N(R x1 )C(O)N(R x1a )(R x1b ), -OC(O)N(R x1 )(R x1a ), -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be one or more -R x2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(R x3 )C(O)N(R x3a )- and -OC(O)N(R x3 )-, -R x1 , -R x1a , -R x1b -H, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50Alkynyl may be one or more -R x2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(R x3 )C(O)N(R x3a )- and -OC(O)N(R x3 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R x2 and optionally independently substituted with Each-R x2 is halogen, -CN, oxo(=O), -C(O)OR x4 , -OR x4 , -C(O)R x4 , -C(O)N(R x4 )(R x4a ), -S(O)2N(R x4 )(R x4a ), -S(O)N(R x4 )(R x4a ), -S(O)R x4 , -S(O)R x4 , -N(R x4 )S(O)2N(R x4a )(R x4b ), -SR x4 , -N(R x4 )(R x4a), -NO2, -OC(O)R x4 , -N(R x4 )C(O)R x4a , -N(R x4 )S(O)2R x4a , -N(R x4 )S(O)R x4a , -N(R x4 )C(O)OR x4a , -N(R x4 )C(O)N(R x4a )(R x4b ), -OC(O)N(R x4 )(R x4a ) and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R x3 , -R x3a , -R x4 , -R x4a , -R x4b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different.

[0078] In certain embodiments, the term "substituents" includes halogen, -CN, -C(O)OR x1 , -OR x1 , -C(O)R x1 , -C(O)N(R x1 )(R x1a ), -S(O)2N(R x1 )(R x1a ), -S(O)N(R x1 )(R x1a ), -S(O)R x1 , -S(O)R x1 , -N(R x1 )S(O)2N(R x1 )(R x1a ), -SR x1 , -N(R x1 )(R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a, -N(R x1 )S(O)2R x1a , -N(R x1 )S(O)R x1a , -N(R x1 )C(O)OR x1a , -N(R x1 )C(O)N(R x1 )(R x1a ), -OC(O)N(R x1 )(R x1a ), -T 0 , C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl, -T 0 , C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 Alkynyl may be one or more -R x2 is optionally replaced by C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(R x3 )C(O)N(R x3a )- and -OC(O)N(R x3 )-, Each-R x1 , -R x1a , -R x1b , -R x3 , -R x3a -H, halogen, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl; Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R x2 and optionally independently substituted with Each-R x2 is halogen, -CN, oxo(=O), -C(O)OR x4 , -OR x4 , -C(O)R x4 , -C(O)N(R x4 )(R x4a ), -S(O)2N(R x4 )(R x4a ), -S(O)N(R x4 )(R x4a ), -S(O)R x4 , -S(O)R x4 , -N(R x4 )S(O)2N(R x4a )(R x4b ), -SR x4 , -N(R x4 )(R x4a ), -NO2, -OC(O)R x4 , -N(R x4 )C(O)R x4a , -N(R x4 )S(O)2R x4a , -N(R x4 )S(O)R x4a , -N(R x4 )C(O)OR x4a , -N(R x4 )C(O)N(R x4a )(R x4b ), -OC(O)N(R x4 )(R x4a ) and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R x4 , -R x4a , -R x4b -H, halogen, C 1~6Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl.

[0079] In certain embodiments, the term "substituents" includes halogen, -CN, -C(O)OR x1 , -OR x1 , -C(O)R x1 , -C(O)N(R x1 )(R x1a ), -S(O)2N(R x1 )(R x1a ), -S(O)N(R x1 )(R x1a ), -S(O)R x1 , -S(O)R x1 , -N(R x1 )S(O)2N(R x1a )(R x1b ), -SR x1 , -N(R x1 )(R x1a ), -NO2, -OC(O)R x1 , -N(R x1 )C(O)R x1a , -N(R x1 )S(O)2R x1a , -N(R x1 )S(O)R x1a , -N(R x1 )C(O)OR x1a , -N(R x1 )C(O)N(R x1a )(R x1b ), -OC(O)N(R x1 )(R x1a ), -T 0 , C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, -T 0 , C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 Alkynyl may be one or more -R x2 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R x3 )-, -S(O)2N(R x3 )-, -S(O)N(R x3 )-, -S(O)2-, -S(O)-, -N(R x3 )S(O)2N(R x3a )-, -S-, -N(R x3 )-, -OC(OR x3 )(R x3a )-, -N(R x3 )C(O)N(R x3a )- and -OC(O)N(R x3 )-, Each-R x1 , -R x1a , -R x1b , -R x2 , -R x3 , -R x3a -H, halogen, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl; Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R x2 are independently and optionally substituted with

[0080] In certain embodiments, up to six -H atoms of an optionally substituted molecule are independently replaced by substituents, e.g., five -H atoms are independently replaced by substituents, four -H atoms are independently replaced by substituents, three -H atoms are independently replaced by substituents, two -H atoms are independently replaced by substituents, or one -H atom is replaced by a substituent.

[0081] As used herein, the term "therapeutically effective amount" means an amount sufficient to cure, alleviate or partially arrest the clinical symptoms of a given disease and its complications. Effective amounts for each purpose will depend on the severity of the disease and the weight and general state of the subject.

[0082] As used herein, the term "subject" refers to an animal. Typically, an animal is a mammal. The term "subject" also refers to, for example, primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, etc. In certain embodiments, the subject is a primate. In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human primate.

[0083] As used herein, a subject is "in need of" or "in need thereof" a treatment if the subject would benefit biologically, medically, or in quality of life from the treatment.

[0084] As used herein, the term "water-insoluble" refers to a compound in which less than 1 gram can be dissolved in 1 liter of water at 20° C. to form a homogeneous solution. Accordingly, the term "water-soluble" refers to a compound in which 1 gram or more can be dissolved in 1 liter of water at 20° C. to form a homogeneous solution.

[0085] As used herein, the term "buffer" or "buffering agent" refers to a chemical compound that maintains the pH of a solution within a desired range.

[0086] As used herein, the term "emulsifying agent" or "emulsifier" refers to a chemical compound, such as a surface-active ingredient, that adsorbs to the newly formed interface between the dispersed and continuous phase solutions during emulsion formation, allowing the solutions to mix and protecting the newly formed droplets from immediate reagglomeration.

[0087] As used herein, the term "pH adjuster" refers to a chemical compound used to alter the pH of the droplets within the emulsion, e.g., the emulsion of step (a), to initiate and / or accelerate the cross-linking reaction between the first functionalized HA and the second functionalized HA.

[0088] As used herein, the term "blocking reagent" refers to a chemical compound used to cap unreacted functional groups such as -FG1 or -FG2.

[0089] As used herein, the term "flow system" or "continuous flow system" refers to a system in which a process, such as polymer precipitation, is carried out in a continuously flowing stream rather than in a batch production.

[0090] As used herein, the term "mechanism for precipitating and isolating a polymer" refers to equipment or a device that includes a flow system connected to a collection assembly.

[0091] As used herein, the term "poor solvent" refers to a solvent in which a polymer, such as functionalized HA, is insoluble. The term "insoluble" in relation to a polymer means that less than 1 gram of the polymer can be dissolved in 1 liter of the solvent at room temperature (room temperature can range from 17°C to 30°C, e.g., 17°C to 25°C) to form a homogeneous solution.

[0092] As used herein, "screen scroll centrifuge" refers to a filtering centrifuge that separates solids and liquids from a solid-liquid mixture. In a typical screen scroll centrifuge, the basic principle is that the incoming feed material is separated into two products: liquid and solid.

[0093] In general, the terms "comprise(s)" or "comprising" also encompass "consist(s) of" or "consisting of".

[0094] The drug conjugate of the present invention or a pharmaceutically acceptable salt thereof has a Z in the range of about 50% to about 98%. 1 , Z in the range of about 0.1% to about 20% 2 , Z in the range of about 0.1% to about 20% 3 and Z in the range of about 0.1% to about 10% 4 may include:

[0095] In certain embodiments, the drug conjugate or pharmaceutically acceptable salt thereof has a Z in the range of about 75% to about 98%. 1 , Z in the range of about 0.1% to about 10% 2 , Z in the range of about 0.1% to about 10% 3 and Z in the range of about 0.1% to about 5% 4 Includes.

[0096] In certain embodiments, the drug conjugate or pharmaceutically acceptable salt thereof has a Z in the range of about 78% to about 96%. 1 , Z in the range of about 2% to about 10% 2 , Z in the range of about 1% to about 7% 3 and Z in the range of about 0.5% to about 5% 4 Includes.

[0097] In certain embodiments, the drug conjugate or pharmaceutically acceptable salt thereof has a Z in the range of about 90.5% to about 94.8%. 1 , Z in the range of about 2.7% to about 6.4% 2 , Z in the range of about 1.1% to about 2.1% 3 and Z in the range of about 0.8% to about 1.9% 4 Includes.

[0098] Preferably, the drug conjugate or pharmaceutically acceptable salt thereof has a Z of about 92.9%. 1 , about 4.3% Z 2 , about 1.5% Z 3 and about 1.3% Z 4 Includes.

[0099] It is understood that the percentages provided above are calculated based on the total number of units present in the drug conjugate.

[0100] It will be understood by those skilled in the art that the drug conjugates or pharmaceutically acceptable salts thereof of the present invention may contain small amounts of other HA units that may result from various intramolecular reactions, such as HA units in which -D acts as a crosslinker between two functionalized HA chains. Furthermore, the drug conjugates or pharmaceutically acceptable salts thereof may contain small amounts of HA units that have undergone degradation or other modifications.

[0101] In certain embodiments, all moieties-D of a drug conjugate or a pharmaceutically acceptable salt thereof are identical, i.e., have the same chemical structure. In such cases, all moieties-D of the drug conjugate are derived from the same type of drug molecule.

[0102] In certain embodiments, the drug conjugates of the present invention, or pharmaceutically acceptable salts thereof, comprise different moieties-D, i.e., moieties-D with different chemical structures. These different structures are derived from different types of drug molecules. In certain embodiments, the drug conjugates of the present invention comprise two different types of moieties-D. In certain embodiments, the drug conjugates of the present invention comprise three different types of moieties-D. In certain embodiments, the drug conjugates of the present invention comprise four different types of moieties-D. In certain embodiments, the drug conjugates of the present invention comprise five different types of moieties-D.

[0103] When the drug conjugate of the invention comprises more than one type of -D, all moieties -D are the same type of -L 1 - or a different type of -L 1 -, i.e., a first type of -D can be conjugated to a first type of -L 1 -, and the second type of -D can be conjugated to a second type of -L 1 -, etc. Different types of -L 1The use of -L in certain embodiments allows for different release rates for the different types of -D, e.g., faster release for the first type of -D, medium release for the second type of -D, and slow release for the third type of -D. Thus, in certain embodiments, the drug conjugates of the present invention comprise one type of -L 1 In certain embodiments, the drug conjugates of the invention comprise two types of -L 1 In certain embodiments, the drug conjugates of the invention comprise three types of -L 1 In certain embodiments, the drug conjugates of the invention comprise four types of -L 1 -Includes.

[0104] In certain embodiments, the drug conjugate of the invention comprises one -D and one -L 1 In certain embodiments, the drug conjugates of the invention comprise two -D and two -L 1 In certain embodiments, the drug conjugates of the invention comprise three -D and three -L 1 In certain embodiments, the drug conjugates of the invention comprise four -D and four -L 1 -Includes.

[0105] In certain embodiments, all moieties -L of the drug conjugate 1 - have the same structure. In certain embodiments, the drug conjugate comprises two or more different types of -L 1 -, for example 2, 3, 4 or 5 different types of -L 1 -Contains two or more different types of -L 1 - may be conjugated to the same or different types of -D. Different types of -L 1 -, for example, by using a moiety of the first group -L 1 -L of the second group, which has a long half-life 1It is possible to release the same or different types of drugs DH or -D from the conjugates of the invention with different release half-lives, for example when combined with -.

[0106] In certain embodiments, the VEGF-neutralizing drug moiety (-D) is selected from the group consisting of a soluble VEGF-neutralizing drug moiety, a VEGF-downregulating drug moiety, and a VEGFR-inhibiting drug moiety.

[0107] Exemplary soluble VEGF-neutralizing drug moieties may be selected from the group consisting of anti-VEGF antibodies and anti-VEGF antibody fragments, e.g., bispecific antibodies, fab, scFv, nanobodies, VHHs, anti-VEGF antibody fusion proteins; anti-VEGF antibody-like ligand binding agents, e.g., DARPins, aptamers, adnectins, affibodies, affimers, anticalins, avimers, finomers, knottins, Kunitz domains, abdurins, affitins, anticalins, bicyclic peptides, monobodies; soluble VEGF multivalent binding agents, e.g., diabodies, minibodies, dutafab, crossmab, DARTs, TRIDENTs, DVD-Ig, heterodimeric Fvs, TandAbs, XmAbs, pentamers, hexamers, hexabodies, and soluble VEGF receptor decoy drug moieties that prevent VEGF from binding to its cognate receptor.

[0108] Exemplary soluble VEGF downregulating drug moieties may be selected from the group consisting of shRNA, siRNA, miRNA, aptamers, anti-miRNA, mRNA, antisense RNA, antisense oligonucleotides, plasmids, ribozymes and small molecule drug moieties.

[0109] In certain embodiments, the siRNA drug moiety is selected from the group consisting of Bevasiranib and AGN-745.

[0110] Exemplary VEGFR inhibitor drug moieties may be selected from the group consisting of VEGFR receptor and tyrosine kinase inhibitor drug moieties.

[0111] Exemplary tyrosine kinase inhibitor drug moieties may be selected from the group consisting of vatalanib, linifanib, axitinib, regorafenib, nintedanib, lenvatinib, cabozantinib, anlotinib, and SAR131675.

[0112] In certain embodiments, the VEGFR receptor inhibitor drug moiety is ramucirumab.

[0113] In certain embodiments, the VEGF-neutralizing drug moiety (-D) is an anti-VEGF antibody drug moiety, e.g., an anti-VEGF antibody drug moiety selected from the group consisting of ranibizumab, bevacizumab, brolucizumab, pegaptanib, aflibercept, faricimab, KH902, abicipar pegol, ESBA1008, OPT-302, BI 836880, IBI-302, conbercept, and KSI-301 and / or fragments thereof.

[0114] The CDRs found in the variable regions of antibodies can be defined by the Kabat or IMGT method (Martin, ACR (1996) Accessing the Kabat Antibody Sequence Database by Computer PROTEINS: Structure. Function and Genetics 25: 130-133; Johnson, G. and Wu, TT (2004) The Kabat Database and a Bioinformatics Example. Methods in Molecular Biology 248: 11-25; MacCallum, RM, Martin, ACR and Thornton, JT (1996) Antibody-antigen interactions: Contact analysis and binding site topography. J. Mol. Biol. 262, 732-745; Lefranc, M.-P., Pommie, C., Ruiz, M., Giudicelli, V., Foulquier, E., Truong, L., Thouvenin-Contet, V. and Lefranc, G. (2003) IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains. Dev. Comp. Immunol. 27: 55-77; and Lefranc, M.-P. (2005) IMGT, the international ImMunoGeneTics information System(R). Nucleic Acids Res. 33: D593-D597; Lefranc, M.-P. et al. (2009) IMGT(R), the international ImMunoGeneTics information System(R). Nucleic Acids Res. 37: D1006-D1012).The specificity of an antibody can be defined or described by its set of CDRs, which can be defined by several methods, including the Kabat method or the IMGT® method.

[0115] In certain embodiments, the light chain of the anti-VEGF antibody drug moiety comprises the following three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3): CDR-L1: SASQDISNYLN (SEQ ID NO: 1) CDR-L2: FTSSLHS (SEQ ID NO: 2) CDR-L3: QQYSTVPWT (SEQ ID NO: 3).

[0116] In certain embodiments, the light chain of the anti-VEGF antibody drug moiety comprises the following three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3): CDR-H1: GYDFTHYGMN (SEQ ID NO: 4) CDR-H2: WINTYTGEPTYAADFKR (SEQ ID NO: 5) CDR-H3: YPYYYGTSHWYFDV (SEQ ID NO: 6).

[0117] In certain embodiments, the anti-VEGF antibody drug moiety comprises the following three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) and three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3): CDR-L1: SASQDISNYLN (SEQ ID NO: 1) CDR-L2: FTSSLHS (SEQ ID NO: 2) CDR-L3: QQYSTVPWT (SEQ ID NO: 3) CDR-H1: GYDFTHYGMN (SEQ ID NO: 4) CDR-H2: WINTYTGEPTYAADFKR (SEQ ID NO: 5) CDR-H3: YPYYYGTSHWYFDV (SEQ ID NO: 6).

[0118] In certain embodiments, the light chain of the anti-VEGF antibody drug moiety comprises the following three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) based on Kabat numbering: CDR-L1 (Kabat): SASQDISNYLN (SEQ ID NO: 1) CDR-L2 (Kabat): FTSSLHS (SEQ ID NO: 2) CDR-L3 (Kabat): QQYSTVPWT (SEQ ID NO: 3).

[0119] In certain embodiments, the light chain of the anti-VEGF antibody drug moiety comprises the following three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) based on Kabat numbering: CDR-H1 (Kabat): HYGMN (SEQ ID NO: 7) CDR-H2 (Kabat): WINTYTGEPTYAADFKR (SEQ ID NO: 5) CDR-H3 (Kabat): YPYYYGTSHWYFDV (SEQ ID NO: 6).

[0120] In certain embodiments, the anti-VEGF antibody drug moiety comprises the following three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) and three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) based on Kabat numbering: CDR-L1 (Kabat): SASQDISNYLN (SEQ ID NO: 1) CDR-L2 (Kabat): FTSSLHS (SEQ ID NO: 2) CDR-L3 (Kabat): QQYSTVPWT (SEQ ID NO: 3) CDR-H1 (Kabat): HYGMN (SEQ ID NO: 7) CDR-H2 (Kabat): WINTYTGEPTYAADFKR (SEQ ID NO: 5) CDR-H3 (Kabat): YPYYYGTSHWYFDV (SEQ ID NO: 6).

[0121] In certain embodiments, the light chain of the anti-VEGF antibody drug moiety comprises the following three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) based on IMGT numbering: CDR-L1 (IMGT): QDISNY (SEQ ID NO: 8) CDR-L2(IMGT):FTS CDR-L3(IMGT):QQYSTVPWT (SEQ ID NO:9).

[0122] In certain embodiments, the light chain of the anti-VEGF antibody drug moiety comprises the following three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) based on IMGT numbering: CDR-H1 (IMGT): GYDFTHYG (SEQ ID NO: 10) CDR-H2 (IMGT): INTYTGEP (SEQ ID NO: 11) CDR-H3(IMGT): AKYPYYYGTSHWYFDV (SEQ ID NO: 12).

[0123] In certain embodiments, the anti-VEGF antibody drug moiety comprises the following three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) and three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) based on IMGT numbering: CDR-L1 (IMGT): QDISNY (SEQ ID NO: 8) CDR-L2(IMGT):FTS CDR-L3 (IMGT): QQYSTVPWT (SEQ ID NO: 9) CDR-H1 (IMGT): GYDFTHYG (SEQ ID NO: 10) CDR-H2 (IMGT): INTYTGEP (SEQ ID NO: 11) CDR-H3(IMGT): AKYPYYYGTSHWYFDV (SEQ ID NO: 12).

[0124] In certain embodiments, the anti-VEGF antibody drug moiety comprises the light chain variable domain sequence of SEQ ID NO: 13: DIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKRTV (SEQ ID NO: 13).

[0125] In certain embodiments, the anti-VEGF antibody drug moiety comprises the heavy chain variable domain sequence of SEQ ID NO: 14: EVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTL (SEQ ID NO: 14).

[0126] In certain embodiments, the anti-VEGF antibody drug moiety comprises the light chain variable domain sequence of SEQ ID NO:13 and the heavy chain variable domain sequence of SEQ ID NO:14.

[0127] In certain embodiments, the anti-VEGF antibody drug moiety comprises the light chain of SEQ ID NO: 15: DIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 15).

[0128] In certain embodiments, the anti-VEGF antibody drug moiety comprises a heavy chain of SEQ ID NO: 16: EVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHL (SEQ ID NO: 16).

[0129] Preferably, the anti-VEGF antibody drug moiety is a ranibizumab moiety. Ranibizumab comprises a light chain of SEQ ID NO:15 and a heavy chain of SEQ ID NO:16.

[0130] Any reference herein to a biological drug, i.e., a drug manufactured in, extracted from, or semi-synthesized from a biological source, such as a protein drug, also extends to biosimilar forms of said drug. More specifically, any reference to ranibizumab also extends to its biosimilars, such as Xlucane™, Biooviz™, Razumab™, RanizRel™, and Cimerli™.

[0131] In certain embodiments, each -X'- and -Y'- is independently -T'-, C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl, and C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be one or more -R y1 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y2 )-, -S(O)2N(R y2 )-, -S(O)N(R y2 )-, -S(O)2-, -S(O)-, -N(R y2 )S(O)2N(R y2a )-, -S-, -N(R y2 )-, -OC(OR y2 )(R y2a )-, -N(R y2 )C(O)N(R y2a )- and -OC(O)N(R y2 )-, Each -T'- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; each -T'- is the same or different and is selected from the group consisting of one or more -R y1 and optionally independently substituted with Each-R y1 is halogen, -CN, oxo(=O), -COOR y3 , -OR y3 , -C(O)R y3 , -C(O)N(R y3 R y3a ), -S(O)2N(R y3 R y3a ), -S(O)N(R y3 R y3a ), -S(O)R y3 , -S(O)R y3 , -N(R y3 )S(O)2N(R y3a R y3b ), -SR y3 , -N(R y3 R y3a ), -NO2, -OC(O)R y3 , -N(R y3 )C(O)R y3a , -N(R y3 )S(O)2R y3a , -N(R y3 )S(O)R y3a , -N(R y3 )C(O)OR y3a , -N(R y3 )C(O)N(R y3a R y3b ), -OC(O)N(R y3 R y3a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R y2 , -R y2a , -R y3 , -R y3a , -R y3b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6The alkyl is optionally substituted with one or more halogens which may be the same or different.

[0132] In certain embodiments, each -X'- and -Y'- is independently -T'-, C 1~25 Alkyl, C 2~25 Alkenyl, and C 2~25 alkynyl, and C 1~25 Alkyl, C 2~25 Alkenyl and C 2~25 Alkynyl may be one or more -R y1 is optionally replaced by C 1~25 Alkyl, C 2~25 Alkenyl, and C 2~25 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y2 )-, -S(O)2N(R y2 )-, -S(O)N(R y2 )-, -S(O)2-, -S(O)-, -N(R y2 )S(O)2N(R y2a )-, -S-, -N(R y2 )-, -OC(OR y2 )(R y2a )-, -N(R y2 )C(O)N(R y2a )- and -OC(O)N(R y2 )-, Each -T'- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; each -T'- is the same or different and is selected from the group consisting of one or more -R y1 and optionally independently substituted with Each-R y1 is halogen, -CN, oxo(=O), -COOR y3 , -OR y3 , -C(O)R y3 , -C(O)N(R y3 Ry3a ), -S(O)2N(R y3 R y3a ), -S(O)N(R y3 R y3a ), -S(O)R y3 , -S(O)R y3 , -N(R y3 )S(O)2N(R y3a R y3b ), -SR y3 , -N(R y3 R y3a ), -NO2, -OC(O)R y3 , -N(R y3 )C(O)R y3a , -N(R y3 )S(O)2R y3a , -N(R y3 )S(O)R y3a , -N(R y3 )C(O)OR y3a , -N(R y3 )C(O)N(R y3a R y3b ), -OC(O)N(R y3 R y3a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R y2 , -R y2a , -R y3 , -R y3a , -R y3b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different.

[0133] In certain embodiments, each -X'- and -Y'- is independently -T'-, C 1~18 Alkyl, C 2~18 Alkenyl, and C 2~18 alkynyl, and C 1~18 Alkyl, C 2~18 Alkenyl and C 2~18Alkynyl may be one or more -R y1 is optionally replaced by C 1~18 Alkyl, C 2~18 Alkenyl, and C 2~18 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y2 )-, -S(O)2N(R y2 )-, -S(O)N(R y2 )-, -S(O)2-, -S(O)-, -N(R y2 )S(O)2N(R y2a )-, -S-, -N(R y2 )-, -OC(OR y2 )(R y2a )-, -N(R y2 )C(O)N(R y2a )- and -OC(O)N(R y2 )-, Each -T'- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; each -T'- is the same or different and is selected from the group consisting of one or more -R y1 and optionally independently substituted with Each-R y1 is halogen, -CN, oxo(=O), -COOR y3 , -OR y3 , -C(O)R y3 , -C(O)N(R y3 R y3a ), -S(O)2N(R y3 R y3a ), -S(O)N(R y3 R y3a ), -S(O)R y3 , -S(O)R y3 , -N(R y3 )S(O)2N(R y3a R y3b ), -SR y3 , -N(R y3 R y3a ), -NO2, -OC(O)Ry3 , -N(R y3 )C(O)R y3a , -N(R y3 )S(O)2R y3a , -N(R y3 )S(O)R y3a , -N(R y3 )C(O)OR y3a , -N(R y3 )C(O)N(R y3a R y3b ), -OC(O)N(R y3 R y3a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R y2 , -R y2a , -R y3 , -R y3a , -R y3b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different.

[0134] In certain embodiments, each -X'- and -Y'- is independently -T'-, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl, and C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 Alkynyl may be one or more -R y1 is optionally replaced by C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y2 )-, -S(O)2N(R y2 )-, -S(O)N(R y2 )-, -S(O)2-, -S(O)-, -N(R y2 )S(O)2N(R y2a)-, -S-, -N(R y2 )-, -OC(OR y2 )(R y2a )-, -N(R y2 )C(O)N(R y2a )- and -OC(O)N(R y2 )-, Each -T'- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; each -T'- is the same or different and is selected from the group consisting of one or more -R y1 and optionally independently substituted with Each-R y1 is halogen, -CN, oxo(=O), -COOR y3 , -OR y3 , -C(O)R y3 , -C(O)N(R y3 R y3a ), -S(O)2N(R y3 R y3a ), -S(O)N(R y3 R y3a ), -S(O)R y3 , -S(O)R y3 , -N(R y3 )S(O)2N(R y3a R y3b ), -SR y3 , -N(R y3 R y3a ), -NO2, -OC(O)R y3 , -N(R y3 )C(O)R y3a , -N(R y3 )S(O)2R y3a , -N(R y3 )S(O)R y3a , -N(R y3 )C(O)OR y3a , -N(R y3 )C(O)N(R y3a R y3b ), -OC(O)N(R y3 R y3a ), and C1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R y2 , -R y2a , -R y3 , -R y3a , -R y3b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different.

[0135] In certain embodiments, each -X'- and -Y'- is independently -T'-, C 1~5 Alkyl, C 2~5 Alkenyl, and C 2~5 alkynyl, and C 1~5 Alkyl, C 2~5 Alkenyl and C 2~5 Alkynyl may be one or more -R y1 is optionally replaced by C 1~5 Alkyl, C 2~5 Alkenyl, and C 2~5 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y2 )-, -S(O)2N(R y2 )-, -S(O)N(R y2 )-, -S(O)2-, -S(O)-, -N(R y2 )S(O)2N(R y2a )-, -S-, -N(R y2 )-, -OC(OR y2 )(R y2a )-, -N(R y2 )C(O)N(R y2a )- and -OC(O)N(R y2 )-, Each -T'- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; each -T'- is the same or different and is selected from the group consisting of one or more -R y1 and optionally independently substituted with Each-R y1 is halogen, -CN, oxo(=O), -COOR y3 , -OR y3 , -C(O)R y3 , -C(O)N(R y3 R y3a ), -S(O)2N(R y3 R y3a ), -S(O)N(R y3 R y3a ), -S(O)R y3 , -S(O)R y3 , -N(R y3 )S(O)2N(R y3a R y3b ), -SR y3 , -N(R y3 R y3a ), -NO2, -OC(O)R y3 , -N(R y3 )C(O)R y3a , -N(R y3 )S(O)2R y3a , -N(R y3 )S(O)R y3a , -N(R y3 )C(O)OR y3a , -N(R y3 )C(O)N(R y3a R y3b ), -OC(O)N(R y3 R y3a ), and C 1~4 alkyl, independently selected from the group consisting of C 1~4 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R y2 , -R y2a , -R y3 , -R y3a , -R y3b -H and C 1~4 alkyl, independently selected from the group consisting of C 1~4The alkyl is optionally substituted with one or more halogens which may be the same or different.

[0136] In certain embodiments, -X'- is a group of formula (x0):

[0137] [ka] [In the formula, The unmarked dashed line indicates a bond to -X-, and the dashed line with an asterisk indicates a bond to -FG1 or -L. 3 - indicates a bond to v0 is selected from the group consisting of 0 and 1; -X 1 -, -X 4 - is independently C 1~10 This C is an alkyl 1~10 Alkyl is -O-, -T-, -N(R y1 )-, -C(O)O- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1~10 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a Optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -R y2a is H and C 1~4 independently selected from the group consisting of alkyl, -X 2 - is -N(R 1 )—, —O—, —S—, and —Se—; =X 3 is =O, =N(R 1 ) and =S; -X 5 -C 1~20 This C is an alkyl 1~20 Alkyl is -O-, -C(O)O-, -T-, -N(R y1 )- and -N(R y1)C(O)—, wherein the C 1~20 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a Optionally substituted with one or more groups independently selected from -R 1 -H, C 1~5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 2 and optionally independently substituted with -R 2 is halogen, -CN, oxo, -C(O)OR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 )(R 3a ), -S(O)2N(R 3 )(R 3a ), -S(O)N(R 3 )(R 3a ), -S(O)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a )(R 3b ), -SR 3 , -N(R 3 )(R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 3a , -N(R 3 )S(O)R 3a , -N(R 3 )C(O)OR 3a , -N(R 3 )C(O)N(R 3a )(R 3b ), -OC(O)N(R3 )(R 3a ) and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 3 , -R 3a and -R 3b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. It is of the type.

[0138] In certain embodiments, v0 in formula (x0) is 0. In certain embodiments, v0 in formula (x0) is 1.

[0139] In certain embodiments, -X'- is a group represented by formula (x1):

[0140] [ka] [In the formula, The unmarked dashed line indicates a bond to -X-, and the dashed line with an asterisk indicates a bond to -FG1 or -L. 3 - indicates a bond to b0 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; -X 1 -, -X 4 - is independently C 1~5 This C is an alkyl 1~5 Alkyl is -O-, -T-, -N(R y1 )- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1~5 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a ) optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -Ry2a -H and C 1~4 independently selected from the group consisting of alkyl, -X 2 - is -N(R 1 )—, —O—, —S—, and —Se—; =X 3 is =O, =N(R 1 ) and =S; -X 6 -C 1~10 This C is an alkyl 1~10 Alkyl is -O-, -C(O)O-, -T-, -N(R y1 )- and -N(R y1 )C(O)—, wherein the C 1~10 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a Optionally substituted with one or more groups independently selected from -R 1 -H, C 1~5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 2 and optionally independently substituted with -R 2 is halogen, -CN, oxo, -C(O)OR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 )(R 3a ), -S(O)2N(R 3 )(R 3a ), -S(O)N(R 3 )(R 3a ), -S(O)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R3a )(R 3b ), -SR 3 , -N(R 3 )(R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 3a , -N(R 3 )S(O)R 3a , -N(R 3 )C(O)OR 3a , -N(R 3 )C(O)N(R 3a )(R 3b ), -OC(O)N(R 3 )(R 3a ) and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 3 , -R 3a and -R 3b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. It is of the type.

[0141] In certain embodiments, -X'- is a group represented by formula (x2):

[0142] [ka] [In the formula, The unmarked dashed line indicates a bond to -X-, and the dashed line with an asterisk indicates a bond to -FG1 or -L. 3 - indicates a bond to b0 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; -X 1 -, -X 4 -, -X 7 -, -X 8 - is independently C 1~5This C is an alkyl 1~5 Alkyl is -O-, -T-, -N(R y1 )- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1~5 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a ) optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -R y2a is H and C 1~4 independently selected from the group consisting of alkyl, -R 1 , -R 5 , -R 10 -H, C 1~5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 6 and optionally independently substituted with -R 6 is halogen, -CN, oxo, -C(O)OR 7 , -OR 7 , -C(O)R 7 , -C(O)N(R 7 )(R 7a ), -S(O)2N(R 7 )(R 7a ), -S(O)N(R 7 )(R 7a ), -S(O)R 7 , -S(O)R 7 , -N(R 7 )S(O)2N(R 7a )(R 7b ), -SR 7 , -N(R 7 )(R 7a ), -NO2, -OC(O)R 7 , -N(R 7 )C(O)R7a , -N(R 7 )S(O)2R 7a , -N(R 7 )S(O)R 7a , -N(R 7 )C(O)OR 7a , -N(R 7 )C(O)N(R 7a )(R 7b ), -OC(O)N(R 7 )(R 7a ) and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 7 , -R 7a and -R 7b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. It is of the type.

[0143] In certain embodiments, -X'- is a group represented by formula (x3):

[0144] [ka] [In the formula, The unmarked dashed line indicates a bond to -X-, and the dashed line with an asterisk indicates a bond to -FG1 or -L. 3 - indicates a bond to b0 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; -X 1 -, -X 4 - is independently C 1~5 This C is an alkyl 1~5 Alkyl is -O-, -T-, -N(R y1 )- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1~5 The alkyl chain is -OH, -T, -NH(R y1) and -C(O)N(R y2 R y2a ) optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -R y2a is H and C 1~4 independently selected from the group consisting of alkyl, -R 1 , -R 5 , -R 10 -H, C 1~5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 6 and optionally independently substituted with -R 6 is halogen, -CN, oxo, -C(O)OR 7 , -OR 7 , -C(O)R 7 , -C(O)N(R 7 )(R 7a ), -S(O)2N(R 7 )(R 7a ), -S(O)N(R 7 )(R 7a ), -S(O)R 7 , -S(O)R 7 , -N(R 7 )S(O)2N(R 7a )(R 7b ), -SR 7 , -N(R 7 )(R 7a ), -NO2, -OC(O)R 7 , -N(R 7 )C(O)R 7a , -N(R 7 )S(O)2R 7a , -N(R 7 )S(O)R 7a , -N(R 7 )C(O)OR 7a , -N(R 7 )C(O)N(R 7a)(R 7b ), -OC(O)N(R 7 )(R 7a ) and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 7 , -R 7a and -R 7b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. It is of the type.

[0145] In certain embodiments, b0 in formula (x1), (x2), or (x3) is 1. In certain embodiments, b0 in formula (x1), (x2), or (x3) is 2. In certain embodiments, b0 in formula (x1), (x2), or (x3) is 3. In certain embodiments, b0 in formula (x1), (x2), or (x3) is 4. In certain embodiments, b0 in formula (x1), (x2), or (x3) is 5. In certain embodiments, b0 in formula (x1), (x2), or (x3) is 6. In certain embodiments, b0 in formula (x1), (x2), or (x3) is 7. In certain embodiments, b0 in formula (x1), (x2), or (x3) is 8. In certain embodiments, b0 in formula (x1), (x2), or (x3) is 9. In certain embodiments, b0 in formula (x1), (x2), or (x3) is 10.

[0146] In certain embodiments, -X'- is a group represented by formula (x4):

[0147] [ka] [In the formula, The unmarked dashed line indicates a bond to -X-, and the dashed line with an asterisk indicates a bond to -FG1 or -L. 3 -, and c0 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. It is of the type.

[0148] In certain embodiments, c0 in formula (x4) is 1. In certain embodiments, c0 in formula (x4) is 2. In certain embodiments, c0 in formula (x4) is 3. In certain embodiments, c0 in formula (x4) is 4. In certain embodiments, c0 in formula (x4) is 5. In certain embodiments, c0 in formula (x4) is 6. In certain embodiments, c0 in formula (x4) is 7. In certain embodiments, c0 in formula (x4) is 8. In certain embodiments, c0 in formula (x4) is 9. In certain embodiments, c0 in formula (x4) is 10.

[0149] In certain embodiments, -Y'- is a group of formula (y0):

[0150] [ka] [In the formula, The unmarked dashed lines indicate bonds to -Y-, and the dashed lines with stars indicate bonds to -FG2, -L 3 -, -L 4 -or-L 5 - indicates a bond to -Y 1 -, -Y 4 - is independently C 1~10 This C is an alkyl 1~10 Alkyl is -O-, -T-, -N(R y1 )-, -C(O)O- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1~10 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a ) optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -R y2a is H and C 1~4 independently selected from the group consisting of alkyl, -Y2 - is -N(R 2 )—, —O—, —S—, and —Se—; =Y 3 is =O, =N(R 2 ) and =S; -R 1 , -R 2 -H, C 1~5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 3 and optionally independently substituted with -R 3 is halogen, -CN, oxo, -C(O)OR 4 , -OR 4 , -C(O)R 4 , -C(O)N(R 4 )(R 4a ), -S(O)2N(R 4 )(R 4a ), -S(O)N(R 4 )(R 4a ), -S(O)R 4 , -S(O)R 4 , -N(R 4 )S(O)2N(R 4a )(R 4b ), -SR 4 , -N(R 4 )(R 4a ), -NO2, -OC(O)R 4 , -N(R 4 )C(O)R 4a , -N(R 4 )S(O)2R 4a , -N(R 4 )S(O)R 4a , -N(R 4 )C(O)OR 4a , -N(R 4 )C(O)N(R 4a )(R 4b ), -OC(O)N(R 4 )(R4a ) and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 4 , -R 4a and -R 4b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. It is of the type.

[0151] In certain embodiments, -X'- is of formula (y0), where the unmarked dashed line indicates the bond to -X- and the dashed line with an asterisk indicates the bond to -FG1.

[0152] In certain embodiments, -Y in formula (y0) 2 - is -N(R 2 In certain embodiments, -Y in formula (y0) is 2 - is -O-. In certain embodiments, -Y in formula (y0) 2 - is -S-. In certain embodiments, -Y in formula (y0) 2 - is -Se-.

[0153] In certain embodiments, =Y in formula (y0) 3 In certain embodiments, the =Y in formula (y0) is ═O. 3 is =N(R 2 In certain embodiments, the =Y 3 is =S.

[0154] In certain embodiments, -Y'- is a group represented by the formula (y1):

[0155] [ka] [In the formula, The unmarked dashed lines indicate bonds to -Y-, and the dashed lines with stars indicate bonds to -FG2, -L3 -, -L 4 -or-L 5 - indicates a bond to -Y 1 -, -Y 4 - is independently C 1~10 This C is an alkyl 1~10 Alkyl is -O-, -T-, -N(R y1 )-, -C(O)O- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1~10 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a ) optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -R y2a is H and C 1~4 independently selected from the group consisting of alkyl, -Y 2 - is -N(R 2 )—, —O—, —S—, and —Se—; -R 1 , -R 2 -H, C 1~5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 3 and optionally independently substituted with -R 3 is halogen, -CN, oxo, -C(O)OR 4 , -OR 4 , -C(O)R 4 , -C(O)N(R 4 )(R 4a ), -S(O)2N(R 4 )(R 4a ), -S(O)N(R 4 )(R 4a ), -S(O)R4 , -S(O)R 4 , -N(R 4 )S(O)2N(R 4a )(R 4b ), -SR 4 , -N(R 4 )(R 4a ), -NO2, -OC(O)R 4 , -N(R 4 )C(O)R 4a , -N(R 4 )S(O)2R 4a , -N(R 4 )S(O)R 4a , -N(R 4 )C(O)OR 4a , -N(R 4 )C(O)N(R 4a )(R 4b ), -OC(O)N(R 4 )(R 4a ) and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 4 , -R 4a and -R 4b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. It is of the type.

[0156] In certain embodiments, -X'- is of formula (y1), where the unmarked dashed line indicates the bond to -X- and the dashed line with an asterisk indicates the bond to -FG1.

[0157] In certain embodiments, -Y'- is a group represented by formula (y2):

[0158] [ka] [In the formula, The unmarked dashed lines indicate bonds to -Y-, and the dashed lines with stars indicate bonds to -FG2, -L3 -, -L 4 -or-L 5 - indicates a bond to -R 1 , -R 5 is independently selected from the group consisting of —H, methyl, ethyl, propyl, and isopropyl; a1, a2 are independently selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, preferably from the group consisting of 1, 2, 3, 4, 5, 6, 7 and 8, more preferably from the group consisting of 1, 2, 3, 4, 5 and 6, or even more preferably from the group consisting of 1, 2 and 3. It is of the type.

[0159] In certain embodiments, -X'- is of formula (y2), where the unmarked dashed line indicates the bond to -X- and the dashed line with an asterisk indicates the bond to -FG1.

[0160] In certain embodiments, -Y'- is a group represented by formula (y3):

[0161] [ka] [In the formula, The unmarked dashed lines indicate bonds to -Y-, and the dashed lines with stars indicate bonds to -FG2, -L 3 -, -L 4 -or-L 5 - indicates a bond to -R 1 , -R 5 are independently selected from the group consisting of -H, methyl, ethyl, propyl, and isopropyl. It is of the type.

[0162] In certain embodiments, -X'- is of formula (y3), where the unmarked dashed line indicates the bond to -X- and the dashed line with an asterisk indicates the bond to -FG1 or -L 3 - indicates a bond to

[0163] In certain embodiments, -Y'- is a group represented by formula (y4):

[0164] [ka] [In the formula, The unmarked dashed lines indicate bonds to -Y-, and the dashed lines with stars indicate bonds to -FG2, -L 3 -, -L 4 -or-L 5 - indicates a bond to It is of the type.

[0165] In certain embodiments, -X'- is of formula (y4), where the unmarked dashed line indicates the bond to -X- and the dashed line with an asterisk indicates the bond to -FG1 or -L 3 - indicates a bond to

[0166] In certain embodiments, -L 1 - is attached to the lysine residue of -D. In certain embodiments, -L 1 - is attached to the cysteine ​​residue of -D. In certain embodiments, -L 1 - is attached to the histidine residue of -D. In certain embodiments, -L 1 - is attached to the tryptophan residue of -D. In certain embodiments, -L 1 - is attached to the serine residue of -D. In certain embodiments, -L 1 - is attached to the threonine residue of -D. In certain embodiments, -L 1 - is attached to the tyrosine residue of -D. In certain embodiments, -L 1 - is attached to the aspartic acid residue of -D. In certain embodiments, -L 1 - is attached to the glutamic acid residue of -D. In certain embodiments, -L 1 - is attached to the arginine residue of -D.

[0167] In certain embodiments, at least one moiety -L 1 - is attached to the amino acid residue of -D and one or more additional moieties -L 1- is attached to the modifying moiety present in -D.

[0168] Part-L 1 - may be connected to -D through any type of linkage, provided that this linkage is reversible. 1 - is connected to -D through a linkage selected from the group consisting of amide, ester, carbamate, acetal, aminal, imine, oxime, hydrazone, disulfide, and acylguanidine. 1 - is connected to -D through a linkage selected from the group consisting of amide, ester, carbamate, and acylguanidine. These linkages may not themselves be reversible, but reversibility is provided by -L 1 It is understood that this may be the effect of certain groups of atoms or moieties present in -.

[0169] In certain embodiments, -L 1 - is connected to -D through an amide linkage. In certain embodiments, -L 1 - is connected to -D through an ester linkage. In certain embodiments, -L 1 - is connected to -D through a carbamate linkage. In certain embodiments, -L 1 - is connected to -D through an acylguanidine.

[0170] In certain embodiments, -L 1 - is connected to -D via the nitrogen of the amine functional group of the side chain of the lysine residue of -D. 1 - is connected to -D through the nitrogen of the amine functional group of the side chain of the lysine residue of -D, for example, -D, and -L 1 The linkage formed between - is an amide.

[0171] In certain embodiments, -L 1- has a structure as disclosed in WO 2009 / 095479 A2, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 - is a group represented by the formula (I):

[0172] [ka] [In the formula, The dashed line indicates the attachment of -D to the nitrogen atom by forming an amide bond; -X- is -C(R 4 R 4a )-, -N(R 4 )-, -O-, -C(R 4 R 4a )-C(R 5 R 5a )-, -C(R 5 R 5a )-C(R 4 R 4a )-, -C(R 4 R 4a )-N(R 6 )-, -N(R 6 )-C(R 4 R 4a )-, -C(R 4 R 4a )-O-, -OC(R 4 R 4a )- or -C(R 7 R 7a )- and X 1 is C or S(O), -X 2 - is -C(R 8 R 8a )- or -C(R 8 R 8a )-C(R 9 R 9a )- and =X 3 is =O, =S, or =N-CN, -R 1 , -R 1a , -R 2 , -R 2a , -R 4 , -R4a , -R 5 , -R 5a , -R 6 , -R 8 , -R 8a , -R 9 , -R 9a -H, and C 1~6 independently selected from the group consisting of alkyl, -R 3 , -R 3a -H, and C 1~6 alkyl, with the proviso that -R 3 , -R 3a If one or both of are other than -H, they are sp 3 provided that it is connected to a bonded N through a hybridized carbon atom, -R 7 is -N(R 10 R 10a ), or -NR 10 -(C=O)-R 11 and -R 7a , -R 10 , -R 10a , -R 11 are, independently of each other, -H or C 1~10 is alkyl, Optionally, Pair-R 1a / -R 4a , -R 1a / -R 5a , -R 1a / -R 7a , -R 4a / -R 5a , -R 8a / -R 9a one or more of the following forms a chemical bond; Optionally, Pair-R 1 / -R 1a , -R 2 / -R 2a , -R 4 / -R 4a , -R 5 / -R 5a , -R 8 / -R 8a , -R 9 / -R 9aOne or more of the following, together with the atoms to which they are attached, form a C 3~10 forming a cycloalkyl or a 3- to 10-membered heterocyclyl; Optionally, Pair-R 1 / -R 4 , -R 1 / -R 5 , -R 1 / -R 6 , -R 1 / -R 7a , -R 4 / -R 5 , -R 4 / -R 6 , -R 8 / -R 9 , -R 2 / -R 3 together with the atoms to which they are attached form ring A, Optionally, -R 3 / -R 3a together with the nitrogen atom to which they are attached form a 3- to 10-membered heterocyclic ring, Ring A is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl. It is of Each-L 1 -Ha-L 2 - and optionally further substituted, provided that the hydrogens marked with an asterisk in formula (I) are not replaced by a substituent.

[0173] In certain embodiments, -L 1 - is of formula (I), where the dashed line indicates the attachment of -D to the nitrogen of the amine of the lysine side chain. 1 - is of formula (I) and the dashed line indicates the bond to the nitrogen of the N-terminal amine of -D.

[0174] In certain embodiments, -L of formula (I) 1 - is not further substituted.

[0175] -R in formula (I) 3 / -R 3a However, when they are combined with the nitrogen atom to form a 3- to 10-membered heterocycle, the atom directly bonded to the nitrogen atom is sp 3 It is understood that only 3- to 10-membered heterocyclic rings can be formed that are hybridized carbon atoms. 3 / -R 3a The 3- to 10-membered heterocycle formed by these together with the nitrogen atom to which they are attached has the following structure:

[0176] [ka] [In the formula, The dashed line indicates -L 1 - indicates the bond to the remainder of the ring contains 3 to 10 atoms, including at least one nitrogen; R # and R ## sp 3 represents a hybridized carbon atom].

[0177] It is also understood that the 3- to 10-membered heterocycle may be further substituted.

[0178] -R in formula (I) 3 / -R 3a Exemplary embodiments of suitable 3- to 10-membered heterocycles formed by, together with the nitrogen atom to which they are attached, are:

[0179] [ka] [In the formula, The dashed line indicates the bond to the rest of the molecule; -R is -H and C 1~6 alkyl].

[0180] -L in formula (I) 1- may be optionally further substituted. In general, as long as the principle of cleavage is not affected, i.e., the hydrogen marked with an asterisk in formula (I) is not replaced, and the moiety of formula (I)

[0181] [ka] The nitrogen remains part of a primary, secondary, or tertiary amine, i.e., -R 3 and -R 3a are each independently -H or sp 3 Any substituent can be used as long as it is connected to -N through a hybridized carbon atom.

[0182] In certain embodiments, -R of formula (I) 1 or -R 1a -L 2 In certain embodiments, -R of formula (I) is substituted with -. 2 or -R 2a -L 2 In certain embodiments, -R of formula (I) is substituted with -. 3 or -R 3a -L 2 In certain embodiments, -R of formula (I) is substituted with -. 4 -L 2 In certain embodiments, -R of formula (I) is substituted with -. 5 or -R 5a -L 2 In certain embodiments, -R of formula (I) is substituted with -. 6 -L 2 In certain embodiments, -R of formula (I) is substituted with -. 7 or -R 7a -L 2 In certain embodiments, -R of formula (I) is substituted with -. 8 or -R 8a -L 2 In certain embodiments, -R of formula (I) is substituted with -. 9 or -R 9a -L 2- is preferably substituted with -R in formula (I). 11 -L 2 - is replaced by

[0183] In certain embodiments, -L 1 has a structure as disclosed in WO 2018 / 193408 A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 - is a group represented by the formula (II):

[0184] [ka] [In the formula, The dashed line indicates the bond to -D, which is a primary amine, secondary amine, or ring nitrogen atom of an azaheteroaryl ring, and -R 1 is —H or C1-C4 alkyl, -R 1a is —H or C1-C4 alkyl, or —CR 1 R 1a combine to form a C3-C6 cycloalk-1,1-diyl, -R 2 is independently selected at each occurrence from C1-C4 alkyl or oxo, or two -R 2 groups combine to form a fused C3-C6 cycloalkyl or spiro C3-C6 cycloalk-1,1-diyl group; a is 0, 1, 2, 3, or 4; -R 3 is —H or C1-C4 alkyl, -R 3a is —H or C1-C4 alkyl, or —CR 3 R 3a combine to form a C3-C6 cycloalk-1,1-diyl, -Y is -C(O)R 4 , -C(O)OR 4 , -C(O)NHR 4 , -C(O)NR 5 R 6 , -SiR5 R 6 R 7 , or -CR 12 R 12a OR 13 and -R 12 is —H or C1-C4 alkyl, -R 12a is —H or C1-C4 alkyl, or —CR 12 R 12a combine to form a C3-C6 cycloalk-1,1-diyl, -R 13 is C1-C4 alkyl or -CHR 12 OR 13 combine to form a 5-, 6-, or 7-membered cyclic ether, -R 4 is C1-C8 alkyl or C3-C7 cycloalkyl, wherein the cycloalkyl is optionally substituted with 0, 1, or 2 independently selected C1-C4 alkyl groups, wherein the alkyl is optionally substituted with C1-C4 alkoxy; -R 5 and -R 6 are each independently selected from C1-C4 alkyl and C3-C6 cycloalkyl; -R 7 is C1-C8 alkyl, C3-C7 cycloalkyl, C1-C8 alkoxy, C3-C7 cycloalkyloxy, heterocycloalkyloxy, or -(OCHR 3 CH2) b O-C1-C4 alkyl, where heterocycloalkyloxy is a 4-7 membered saturated heterocyclic ring having one heteroatom selected from N, O, and S and optionally substituted with 0, 1, or 2 independently selected C1-C4 alkyl groups; b is an integer ranging from 1 to 10; -Z, -CH-L 2 - or -NL 2 -is] It is of -L 1 - is optionally further substituted.

[0185] In certain embodiments, -L 1 is a group of formula (Iia):

[0186] [ka] [In the formula, The dashed line with an asterisk indicates the attachment of -D to the nitrogen atom by forming an amide bond; Unmarked dashed lines are -L 2 - indicates a bond to -R 4 is -CH3, -CH2-O-CH3, -CH2CH3, or -CH(CH3)2] It is of the type.

[0187] In certain embodiments, -L 1 - is of formula (Iia), and -L 2 is a group represented by the formula (Iia'):

[0188] [ka] [In the formula, Unmarked dashed lines are -L 1 - indicates a bond to The dashed line with the star indicates -L 4 - indicates a bond to It is of the type.

[0189] In certain embodiments, -L 1 has a structure as disclosed in WO 2016 / 020373 A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 - is a group represented by the formula (III):

[0190] [ka] [In the formula, The dashed line indicates the attachment of -D to a primary or secondary amine or hydroxyl by forming an amide or ester linkage, respectively; -R 1 , -R 1a , -R 2 , -R 2a , -R 3 and -R 3a -H, -C(R 8 R 8a R 8b ), -C(=O)R 8 , -C≡N, -C(=NR 8 )R 8a , -CR 8 (=CR 8a R 8b ), -C≡CR 8 and -T, -R 4 , -R 5 and -R 5a -H, -C(R 9 R 9a R 9b ) and -T, a1 and a2 are independently 0 or 1; Each-R 6 , -R 6a , -R 7 , -R 7a , -R 8 , -R 8a , -R 8b , -R 9 , -R 9a , -R 9b -H, halogen, -CN, -COOR 10 , -OR 10 , -C(O)R 10 , -C(O)N(R 10 R 10a ), -S(O)2N(R 10 R 10a ), -S(O)N(R 10 R 10a ), -S(O)R 10 , -S(O)R 10 , -N(R 10 )S(O)2N(R 10a R 10b), -SR 10 , -N(R 10 R 10a ), -NO2, -OC(O)R 10 , -N(R 10 )C(O)R 10a , -N(R 10 )S(O)2R 10a , -N(R 10 )S(O)R 10a , -N(R 10 )C(O)OR 10a , -N(R 10 )C(O)N(R 10a R 10b ), -OC(O)N(R 10 R 10a ), -T, C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl, -T, C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 Alkynyl may be one or more -R 11 is optionally replaced by C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 12 )-, -S(O)2N(R 12 )-, -S(O)N(R 12 )-, -S(O)2-, -S(O)-, -N(R 12 )S(O)2N(R 12a )-, -S-, -N(R 12 )-, -OC(OR 12 )(R 12a )-, -N(R 12 )C(O)N(R 12a )- and -OC(O)N(R 12 )-, Each-R 10 , -R 10a , -R 10b -H, -T, C 1~20 Alkyl, C 2~20 Alkenyl, and C2~20 alkynyl, -T, C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 Alkynyl may be one or more -R 11 is optionally replaced by C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 12 )-, -S(O)2N(R 12 )-, -S(O)N(R 12 )-, -S(O)2-, -S(O)-, -N(R 12 )S(O)2N(R 12a )-, -S-, -N(R 12 )-, -OC(OR 12 )(R 12a )-, -N(R 12 )C(O)N(R 12a )- and -OC(O)N(R 12 )-, Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 each T is independently selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and 11 and optionally independently substituted with Each-R 11 is halogen, -CN, oxo(=O), -COOR 13 , -OR 13 , -C(O)R 13 , -C(O)N(R 13 R 13a ), -S(O)2N(R 13 R 13a ), -S(O)N(R 13 R 13a ), -S(O)R 13 , -S(O)R 13 , -N(R 13 )S(O)2N(R 13a R 13b ), -SR13 , -N(R 13 R 13a ), -NO2, -OC(O)R 13 , -N(R 13 )C(O)R 13a , -N(R 13 )S(O)2R 13a , -N(R 13 )S(O)R 13a , -N(R 13 )C(O)OR 13a , -N(R 13 )C(O)N(R 13a R 13b ), -OC(O)N(R 13 R 13a ), and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R 12 , -R 12a , -R 13 , -R 13a , -R 13b -H, and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Optionally, Pair-R 1 / -R 1a , -R 2 / -R 2a , -R 3 / -R 3a , -R 6 / -R 6a , -R 7 / -R 7a One or more of the following, together with the atoms to which they are attached, form a C 3~10 forming a cycloalkyl or a 3- to 10-membered heterocyclyl; Optionally, Pair-R 1 / -R 2 , -R 1 / -R 3 , -R 1 / -R 4 , -R 1 / -R 5 , -R 1 / -R6 , -R 1 / -R 7 , -R 2 / -R 3 , -R 2 / -R 4 , -R 2 / -R 5 , -R 2 / -R 6 , -R 2 / -R 7 , -R 3 / -R 4 , -R 3 / -R 5 , -R 3 / -R 6 , -R 3 / -R 7 , -R 4 / -R 5 , -R 4 / -R 6 , -R 4 / -R 7 , -R 5 / -R 6 , -R 5 / -R 7 , -R 6 / -R 7 together with the atoms to which they are attached form ring A, A is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl. It is of -L 1 -Ha-L 2 - and -L 1 - is optionally further substituted.

[0191] -L in formula (III) 1 The optional further substituents of - are preferably as described above.

[0192] In certain embodiments, -L of formula (III) 1 - is not further substituted.

[0193] In certain embodiments, -L 1 has a structure as disclosed in EP1536334B1, WO 2009 / 009712 A1, WO 2008 / 034122 A1, WO 2009 / 143412 A2, WO 2011 / 082368 A2, and US8618124B2, which are incorporated herein by reference.

[0194] In certain embodiments, -L 1 - has a structure as disclosed in US8946405B2 and US8754190B2, which are incorporated herein by reference in their entireties. Thus, in certain embodiments, -L 1 - is a group represented by the formula (IV):

[0195] [ka] [In the formula, The dashed line indicates a bond to -D through a functional group of -D selected from the group consisting of -OH, -SH, and -NH; m is 0 or 1; -R 1 and -R 2 At least one or both of are -CN, -NO2, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkenyl, optionally substituted alkynyl, -C(O)R 3 , -S(O)R 3 , -S(O)2R 3 , and -SR 4 are independently selected from the group consisting of: -R 1 and -R 2 is selected from the group consisting of: -H, optionally substituted alkyl, optionally substituted arylalkyl, and optionally substituted heteroarylalkyl; -R 3is -H, optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -OR 9 and -N(R 9 )2, -R 4 is selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Each-R 5 is independently selected from the group consisting of -H, optionally substituted alkyl, optionally substituted alkenylalkyl, optionally substituted alkynylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; -R 9 is selected from the group consisting of —H and optionally substituted alkyl; -Y- is absent and -X- is -O- or -S-; or -Y- is -N(Q)CH2- and -X- is -O-; Q is selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Optionally, -R 1 and -R 2 may be taken together to form a 3- to 8-membered ring, Optionally, both -R 9 together with the nitrogen to which they are attached form a heterocyclic ring] It is of -L 1 -Ha-L 2 - and -L 1- is optionally further substituted.

[0196] In the context of formula (IV) only, the terms used have the following meanings:

[0197] The term "alkyl," as used herein, includes straight-chain, branched, or cyclic saturated hydrocarbon groups of 1 to 8 carbons, or, in some embodiments, 1 to 6 or 1 to 4 carbon atoms.

[0198] The term "alkoxy" includes alkyl groups linked to oxygen, including methoxy, ethoxy, isopropoxy, cyclopropoxy, cyclobutoxy, and the like.

[0199] The term "alkenyl" includes non-aromatic unsaturated hydrocarbons containing a carbon-carbon double bond.

[0200] The term "alkynyl" includes non-aromatic unsaturated hydrocarbons containing a carbon-carbon triple bond.

[0201] The term "aryl" includes aromatic hydrocarbon groups of 6 to 18 carbons, preferably 6 to 10 carbons, including groups such as phenyl, naphthyl, and anthracenyl. The term "heteroaryl" includes aromatic rings of 3 to 15 carbons and containing at least one N, O, or S atom, preferably 3 to 7 carbons and containing at least one N, O, or S atom, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl, and the like.

[0202] In some cases, the alkenyl, alkynyl, aryl, or heteroaryl moiety may be attached to the remainder of the molecule through an alkylene linkage. In these situations, the substituent is referred to as alkenylalkyl, alkynylalkyl, arylalkyl, or heteroarylalkyl, indicating that an alkylene moiety is located between the alkenyl, alkynyl, aryl, or heteroaryl moiety and the molecule to which the alkenyl, alkynyl, aryl, or heteroaryl is attached.

[0203] The term "halogen" includes bromo, fluoro, chloro and iodo.

[0204] The term "heterocyclic ring" refers to a 4- to 8-membered aromatic or non-aromatic ring containing 3 to 7 carbon atoms and at least one N, O, or S atom. Examples are piperidinyl, piperazinyl, tetrahydropyranyl, pyrrolidine, and tetrahydrofuranyl, as well as the exemplary groups provided above for the term "heteroaryl."

[0205] When the ring system is optionally substituted, suitable substituents are selected from the group consisting of alkyl, alkenyl, alkynyl, or additional rings, each of which is optionally further substituted. Optional substituents for any of the groups, including those described above, include halo, nitro, cyano, -OR, -SR, -NR2, -OCOR, -NRCOR, -COOR, -CONR2, -SOR, -S2R, -SONR2, -SON2NR2, where each R is independently alkyl, alkenyl, alkynyl, aryl, or heteroaryl, or two R groups, together with the atoms to which they are attached, form a ring.

[0206] In certain embodiments, -L 1 - has a structure as disclosed in WO2013 / 036857A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, -L 1 - is a compound of formula (V):

[0207] [ka] [In the formula, The dashed line indicates the bond to -D through the amine functionality of -D; -R 1 is an optionally substituted C1-C6 straight chain, branched, or cyclic alkyl; an optionally substituted aryl; an optionally substituted heteroaryl; an alkoxy; and -NR 5 2, -R 2 is selected from the group consisting of -H, optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl; -R 3 is selected from the group consisting of -H, optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl; -R 4 is selected from the group consisting of -H, optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl; Each-R 5 are independently selected from the group consisting of -H, optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl, or when taken together, two -R 5 can be cycloalkyl or cycloheteroalkyl] It is of -L 1 -Ha-L 2 - and -L 1 - is optionally further substituted.

[0208] In the context of formula (V) only, the terms used have the following meanings:

[0209] "Alkyl," "alkenyl," and "alkynyl" include straight-chain, branched, or cyclic hydrocarbon groups of 1 to 8 carbons, or 1 to 6 carbons, or 1 to 4 carbons, where alkyl is a saturated hydrocarbon, alkenyl contains one or more carbon-carbon double bonds, and alkynyl contains one or more carbon-carbon triple bonds. Unless otherwise specified, they contain 1 to 6 C atoms.

[0210] "Aryl" includes aromatic hydrocarbon groups of 6 to 18 carbons, preferably 6 to 10 carbons, including groups such as phenyl, naphthyl, and anthracene. "Heteroaryl" includes aromatic rings of 3 to 15 carbons and containing at least one N, O, or S atom, preferably 3 to 7 carbons and containing at least one N, O, or S atom, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl, and the like.

[0211] The term "substituted" refers to an alkyl, alkenyl, alkynyl, aryl, or heteroaryl group containing one or more substituents in place of one or more hydrogen atoms. Substituents generally include halogen (including F, Cl, Br, and I), lower alkyl (including straight-chain, branched, and cyclic), lower haloalkyl (including fluoroalkyl, chloroalkyl, bromoalkyl, and iodoalkyl), OH, lower alkoxy (including straight-chain, branched, and cyclic), SH, lower alkylthio (including straight-chain, branched, and cyclic), amino, alkylamino, dialkylamino, silyl (including alkylsilyl, alkoxysilyl, and arylsilyl), nitro, cyano, carbonyl, carboxylic acid, carboxylic acid ester, carboxylic acid amide, aminocarbonyl, aminoacyl, carbamate, urea, may be selected from thiocarbamates, thioureas, ketones, sulfones, sulfonamides, aryls (including phenyl, naphthyl, and anthracenyl), heteroaryls (including 5-membered heteroaryls such as pyrrole, imidazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, thiadiazole, triazole, oxadiazole, and tetrazole, 6-membered heteroaryls such as pyridine, pyrimidine, pyrazine, and fused heteroaryls such as benzofuran, benzothiophene, benzoxazole, benzimidazole, indole, benzothiazole, benzisoxazole, and benzisothiazole).

[0212] In certain embodiments, -L 1 - has the structure as disclosed in US7585837B2, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, -L 1 is a group represented by the formula (VI):

[0213] [ka] [In the formula, The dashed line indicates the bond to -D through the amine functionality of -D; R 1 and R 2 is hydrogen, alkyl, alkoxy, alkoxyalkyl, aryl, alkaryl, aralkyl, halogen, nitro, -SO3H, -SO2NHR 5 , amino, ammonium, carboxyl, PO3H2, and OPO3H2; R 3 , R 4 , and R 5 are independently selected from the group consisting of hydrogen, alkyl, and aryl. It is of -L 1 -Ha-L 2 - and -L 1 - is optionally further substituted.

[0214] Suitable substituents of formula (VI) are alkyl (e.g., C 1~6 alkyl), alkenyl (e.g., C 2~6 alkenyl), alkynyl (e.g., C 2~6 alkynyl), aryl (e.g., phenyl), heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl (e.g., aromatic 4- to 7-membered heterocycle), or halogen moiety.

[0215] In the context of formula (VI) only, the terms used have the following meanings:

[0216] The terms "alkyl," "alkoxy," "alkoxyalkyl," "aryl," "alkaryl," and "aralkyl" refer to alkyl radicals of 1 to 8, preferably 1 to 4, carbon atoms, such as methyl, ethyl, propyl, isopropyl, and butyl, and aryl radicals of 6 to 10 carbon atoms, such as phenyl and naphthyl. The term "halogen" includes bromo, fluoro, chloro, and iodo.

[0217] In certain embodiments, -L 1- has the structure as disclosed in WO2002 / 089789A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, -L 1 - is a group represented by the formula (VII):

[0218] [ka] [In the formula, The dashed line indicates the bond to -D through the amine functionality of -D; Y1 and Y2 are independently O, S, or NR 7 and R 2 , R 3 , R 4 , R 5 , R 6 and R 7 is hydrogen, C 1~6 Alkyl, C 3~12 Branched alkyl, C 3~8 Cycloalkyl, C 1~6 Substituted alkyl, C 3~8 Substituted cycloalkyl, aryl, substituted aryl, aralkyl, C 1~6 Heteroalkyl, substituted C 1~6 Heteroalkyl, C 1~6 Alkoxy, phenoxy, and C 1~6 independently selected from the group consisting of heteroalkoxy; Ar is a moiety that, when included in formula (VII), forms a polysubstituted aromatic hydrocarbon or polysubstituted heterocyclic group; X is a chemical bond, a moiety that is actively transported into a target cell, a hydrophobic moiety, or a combination thereof; y is 0 or 1. It is of -L 1 -Ha-L 2 - and -L 1 - is optionally further substituted.

[0219] In the context of formula (VII) only, the terms used have the following meanings:

[0220] The term "alkyl" includes, for example, alkoxy, C 3~8 Linear, branched, substituted C, including cycloalkyl or substituted cycloalkyl 1~12 It is understood to include alkyl.

[0221] The term "substituted" shall be understood to include the addition or replacement of one or more atoms contained in a functional group or compound with one or more different atoms.

[0222] Substituted alkyl includes carboxyalkyl, aminoalkyl, dialkylamino, hydroxyalkyl, and mercaptoalkyl, substituted cycloalkyl includes moieties such as 4-chlorocyclohexyl, aryl includes moieties such as naphthyl, substituted aryl includes moieties such as 3-bromo-phenyl, aralkyl includes moieties such as toluyl, heteroalkyl includes moieties such as ethylthiophene, substituted heteroalkyl includes moieties such as 3-methoxythiophene, alkoxy includes moieties such as methoxy, and phenoxy includes moieties such as 3-nitrophenoxy. Halo- is understood to include fluoro, chloro, iodo, and bromo.

[0223] In certain embodiments, -L 1 - is a partial structure of formula (VIII):

[0224] [ka] [In the formula, The dashed line with an asterisk indicates the attachment of -D to the nitrogen atom by forming an amide bond; Unmarked dashed lines are -L 1 - indicates the bond to the rest of the group] Including, -L 1 -Ha-L 2 - and -L 1 - is optionally further substituted.

[0225] In certain embodiments, -L1 - is of formula (VIII), where the dashed line with an asterisk indicates the attachment of -D to the nitrogen atom of the amine of the lysine side chain. 1 - is of formula (VIII) and the dashed line with an asterisk indicates the bond to the nitrogen of the N-terminal amine of -D.

[0226] In certain embodiments, -L of formula (VIII) 1 - is one part - L 2 In certain embodiments, -L of formula (VIII) is substituted with -. 1 - is not further substituted.

[0227] In certain embodiments, -L 1 - is a partial structure of formula (IX):

[0228] [ka] [In the formula, The dashed line with an asterisk indicates the attachment of -D to the nitrogen atom by forming a carbamate bond; Unmarked dashed lines are -L 1 - indicates the bond to the rest of the group] Including, -L 1 -Ha-L 2 - and -L 1 - is optionally further substituted.

[0229] In certain embodiments, -L 1 - is of formula (IX) and the dashed line with an asterisk indicates the attachment of -D to the nitrogen atom of the amine of the lysine side chain.

[0230] In certain embodiments, -L 1 - is of formula (IX) and the dashed line with an asterisk indicates the bond to the nitrogen atom of the N-terminal amine of -D.

[0231] In certain embodiments, -L of formula (IX) 1- is not further substituted.

[0232] In certain embodiments, -L 1 - is a group represented by the formula (IX-a):

[0233] [ka] [In the formula, The dashed line with an asterisk indicates the bond of -D to the nitrogen atom, and the unmarked dashed line indicates the bond of -L 2 - indicates a bond to n is 0, 1, 2, 3, or 4; =Y1 is selected from the group consisting of =O and =S; -Y2- is selected from the group consisting of -O- and -S-; -Y3- is selected from the group consisting of -O- and -S-; -Y4- is -O-, -NR 5 - and -C(R 6 R 6a )-, =Y5 is selected from the group consisting of =O and =S; -R 3 , -R 5 , -R 6 , -R 6a are independently selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethylpropyl; -R 4 is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethylpropyl; -W- is C 3~10Cycloalkyl, 8-30 membered carbopolycyclyl, 3-10 membered heterocyclyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 C optionally interrupted by one or more groups selected from the group consisting of 1~20 is selected from the group consisting of alkyl, -Nu is -N(R 7 R 7a ), -N(R 7 OH), -N(R 7 )-N(R 7a R 7b ), -S(R 7 ), -COOH,

[0234] [ka] is a nucleophile selected from the group consisting of -Ar- is

[0235] [ka] (In the formula, The dashed line indicates -L 1 - indicates the bond to the remainder of -Z 1 - is -O-, -S- and -N(R 7 )-, -Z 2 - is -N(R 7 )-is) is selected from the group consisting of -R 7 , -R 7a , -R 7b -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl] It is of -L 1 - is optionally further substituted.

[0236] In certain embodiments, -L1 - is of formula (IX-a) and the dashed line with an asterisk indicates the attachment of -D to the nitrogen atom of the amine of the lysine side chain.

[0237] In certain embodiments, -L 1 - is of formula (IX-a) and the dashed line with an asterisk indicates the bond to the nitrogen of the N-terminal amine of -D.

[0238] In certain embodiments, -L of formula (IX-a) 1 - is not further substituted.

[0239] In certain embodiments, -L 1 - is a group represented by formula (IX-b):

[0240] [ka] [In the formula, The dashed line with an asterisk indicates the bond of -D to the nitrogen atom, and the unmarked dashed line indicates the bond of -L 2 - indicates a bond to n is 0, 1, 2, 3, or 4; =Y1 is selected from the group consisting of =O and =S; -Y2- is selected from the group consisting of -O- and -S-; -Y3- is selected from the group consisting of -O- and -S-; -Y4- is -O-, -NR 5 - and -C(R 6 R 6a )-, =Y5 is selected from the group consisting of =O and =S; -R 2 , -R 3 , -R 5 , -R 6 , -R 6aare independently selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethylpropyl; -R 4 is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethylpropyl; -W- is C 3~10 Cycloalkyl, 8-30 membered carbopolycyclyl, 3-10 membered heterocyclyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 C optionally interrupted by one or more groups selected from the group consisting of 1~20 is selected from the group consisting of alkyl, -Nu is -N(R 7 R 7a ), -N(R 7 OH), -N(R 7 )-N(R 7a R 7b ), -S(R 7 ), -COOH,

[0241] [ka] is a nucleophile selected from the group consisting of -Ar- is

[0242] [ka] (In the formula, The dashed line indicates -L 1 - indicates the bond to the remainder of -Z 1- is -O-, -S- and -N(R 7 )-, -Z 2 - is -N(R 7 )-is) is selected from the group consisting of -R 7 , -R 7a , -R 7b -H, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl] It is of -L 1 - is optionally further substituted.

[0243] In certain embodiments, -L 1 - is of formula (IX-b), and the dashed line with an asterisk indicates the attachment of -D to the nitrogen atom of the amine of the lysine side chain.

[0244] In certain embodiments, -L 1 - is of formula (IX-b), and the dashed line with an asterisk indicates the bond to the nitrogen atom of the N-terminal amine of -D.

[0245] In certain embodiments, -L in formula (IX-b) 1 - is not further substituted.

[0246] In certain embodiments, the =Y 1 is =O.

[0247] In certain embodiments, -Y in formula (IX-a) and (IX-b) 2 - is -O-.

[0248] In certain embodiments, -Y in formula (IX-a) and (IX-b) 3 - is -O-.

[0249] In certain embodiments, -Y in formula (IX-a) and (IX-b) 4 -, -NR 5 -It is.

[0250] In certain embodiments, the =Y 5 is =O.

[0251] In certain embodiments, n in formulas (IX-a) and (IX-b) is 0 or 1. In certain embodiments, n in formulas (IX-a) and (IX-b) is 0. In certain embodiments, n in formulas (IX-a) and (IX-b) is 1.

[0252] In certain embodiments, -R of formula (IX-b) 2 is selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. 2 is selected from the group consisting of -H, methyl, ethyl, n-propyl, and isopropyl. In certain embodiments, -R 2 is selected from -H, methyl, and ethyl. In certain embodiments, -R in formula (IX-b) 2 is -H.

[0253] In certain embodiments, -R of formula (IX-a) and (IX-b) 3 is selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 3 is selected from the group consisting of -H, methyl, ethyl, n-propyl, and isopropyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 3 is selected from -H, methyl, and ethyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 3 is -H.

[0254] In certain embodiments, each -R of formula (IX-a) and (IX-b) 4 is independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 4 is selected from the group consisting of methyl, ethyl, n-propyl, and isopropyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 4 is selected from methyl and ethyl.

[0255] In certain embodiments, -R of formula (IX-a) and (IX-b) 5 is selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 5 is selected from the group consisting of -H, methyl, ethyl, n-propyl, and isopropyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 5 is selected from methyl and ethyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 5 is methyl.

[0256] In certain embodiments, -R of formula (IX-a) and (IX-b) 6 and -R 6a is independently selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 6 and -R 6a is independently selected from the group consisting of -H, methyl, ethyl, n-propyl, and isopropyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 6 and -R 6a is independently selected from —H, methyl, and ethyl. In certain embodiments, —R6 and -R 6a are both -H.

[0257] In certain embodiments, Ar in formula (IX-a) and (IX-b) is phenyl. In certain embodiments, Ar in formula (IX-a) and (IX-b) is

[0258] [ka] wherein the dashed line indicates the bond of the moieties of formula (IX-a) and (IX-b) to the rest of the moieties. is.

[0259] In certain embodiments, W in formulas (IX-a) and (IX-b) is C 3~10 Cycloalkyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 C optionally interrupted by )- 1~20 In certain embodiments, W in formula (IX-a) and (IX-b) is C 3~10 Cycloalkyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 C optionally interrupted by )- 1~10 In certain embodiments, W in formula (IX-a) and (IX-b) is C 3~10 Cycloalkyl, -C(O)-, -C(O)N(R 7 )-, -O-, -S- and -N(R 7 C optionally interrupted by )- 1~6 In certain embodiments, W in formulas (IX-a) and (IX-b) is

[0260] [ka] [In the formula, The dashed line indicates the bond to the remainder of the moiety of formula (IX-a) or (IX-b), respectively. is.

[0261] In certain embodiments, -Nu in formulas (IX-a) and (IX-b) is -N(R 7 R 7a )

[0262] In certain embodiments, -R of formula (IX-a) and (IX-b) 7 , -R 7a and -R 7b are independently selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 7 , -R 7a and -R 7b are independently selected from —H, methyl, ethyl, n-propyl, and isopropyl. In certain embodiments, —R 7 , -R 7a and -R 7b are independently selected from methyl or ethyl. In certain embodiments, -R in formula (IX-a) and (IX-b) 7 , -R 7a and -R 7b are all methyl.

[0263] In certain embodiments, -L 1 - is a group represented by formula (IX-c):

[0264] [ka] [In the formula, The dashed line with an asterisk indicates the bond of -D to the nitrogen atom, Unmarked dashed lines are -L 2 - indicates a bond to s1 is an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. It is of the type.

[0265] In certain embodiments, -L 1- is of formula (IX-c), and the dashed line with an asterisk indicates the attachment of -D to the nitrogen atom of the amine of the lysine side chain.

[0266] In certain embodiments, -L 1 - is of formula (IX-c), and the dashed line with an asterisk indicates the bond to the nitrogen of the N-terminal amine of -D.

[0267] In certain embodiments, s1 in formula (IX-c) is an integer selected from the group consisting of 1, 2, 3, 4, and 5. In certain embodiments, s1 in formula (IX-c) is 1. In certain embodiments, s1 in formula (IX-c) is 2. In certain embodiments, s1 in formula (IX-c) is 3. In certain embodiments, s1 in formula (IX-c) is 4. In certain embodiments, s1 in formula (IX-c) is 5.

[0268] In certain embodiments, -L 1 - is a group represented by the formula (IX-d):

[0269] [ka] [In the formula, The dashed line with an asterisk indicates the bond of -D to the nitrogen atom, Unmarked dashed lines are -L 2 - indicates a bond to It is of the type.

[0270] In certain embodiments, -L 1 - is of formula (IX-d), where the dashed line with an asterisk indicates the attachment of -D to the nitrogen atom of the amine of the lysine side chain.

[0271] In certain embodiments, -L 1 - is of formula (IX-d), where the dashed line with an asterisk indicates the bond to the nitrogen atom of the N-terminal amine of -D.

[0272] In certain embodiments, -L 1- has a structure as disclosed in WO 2020 / 206358 A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 - is a compound of the formula (X):

[0273] [ka] [In the formula, The unmarked dashed line indicates the bond to -D, The dashed line with the star indicates -L 2 - indicates a bond to n is an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6; -R 1 and -R 2 are independently an electron-withdrawing group, alkyl, or -H, and -R 1 or -R 2 at least one of is an electron-withdrawing group, Each-R 4 are independently C1-C3 alkyl, or two -R 4 form a 3- to 6-membered ring together with the carbon atoms to which they are attached, When -D is a drug moiety attached through an amine, -Y- is absent, or when -D is a drug moiety attached through a phenol, alcohol, thiol, thiophenol, imidazole, or non-basic amine, -Y- is replaced by -N(R 6 )CH2- and -R 6 is an optionally substituted C1-C6 alkyl, an optionally substituted aryl, or an optionally substituted heteroaryl. It is of the type.

[0274] In certain embodiments, n in formula (X) is an integer selected from 1, 2, 3, 4, 5, and 6. In certain embodiments, n in formula (X) is an integer selected from 1, 2, and 3. In certain embodiments, n in formula (X) is an integer from 0, 1, 2, and 3. In certain embodiments, n in formula (X) is 1. In certain embodiments, n in formula (X) is 2. In certain embodiments, n in formula (X) is 3.

[0275] In certain embodiments, -R of formula (X) 1 and -R 2 The electron-withdrawing group is selected from the group consisting of -CN; -NO2; optionally substituted aryl; optionally substituted heteroaryl; optionally substituted alkenyl; optionally substituted alkynyl; -COR 3 , -SOR 3 , or -SO2R 3 (-R 3 is -H, optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -OR 8 or -NR 8 2, and each -R 8 are independently —H or optionally substituted alkyl, or both —R 8 groups taken together with the nitrogen to which they are attached form a heterocyclic ring); or -SR 9 (-R 9 is selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl).

[0276] In certain embodiments, -R of formula (X) 1 and -R 2 In certain embodiments, the electron-withdrawing group in formula (X) is —CN. 1 and -R 2In certain embodiments, the electron-withdrawing group of formula (X) is —NO. 1 and -R 2 is an optionally substituted aryl containing 6 to 10 carbon atoms. 1 and -R 2 is an optionally substituted phenyl, naphthyl, or anthracenyl. 1 and -R 2 is an optionally substituted heteroaryl containing 3 to 7 carbons and at least one N, O, or S atom. 1 and -R 2 is an optionally substituted pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, or indenyl. 1 and -R 2 is an optionally substituted alkenyl containing 2 to 20 carbon atoms. In certain embodiments, the -R of formula (X) 1 and -R 2 is an optionally substituted alkynyl containing 2 to 20 carbon atoms. 1 and -R 2 The electron-withdrawing group is -COR 3 , -SOR 3 , or -SO2R 3 (-R 3 is -H, optionally substituted alkyl containing 1 to 20 carbon atoms, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -OR 8 or -NR 8 2, and each -R 8are independently -H or optionally substituted alkyl containing 1 to 20 carbon atoms, or both -R 8 groups, taken together with the nitrogen to which they are attached, form a heterocyclic ring. In certain embodiments, -R of formula (X) 1 and -R 2 The electron-withdrawing group is -SR 9 and -R 9 is an optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl containing 1 to 20 carbon atoms.

[0277] In certain embodiments, -R of formula (X) 1 or -R 2 At least one of the following is -CN, -SOR 3 or -SO2R 3 In certain embodiments, -R of formula (X) 1 and -R 2 At least one of the groups is -CN or -SO2R 3 In certain embodiments, -R of formula (X) 1 and -R 2 At least one of the groups is -CN or -SO2R 3 and -R 3 is optionally substituted alkyl, optionally substituted aryl, or -NR 8 2. In certain embodiments, -R of formula (X) 1 and -R 2 At least one of is -CN, -SO2N(CH3)2, -SO2CH3, phenyl substituted with -SO2, phenyl substituted with -SO2 and -Cl, -SON2N(CH2CH2)2O, -SO2CH(CH3)2, -SON2N(CH3)(CH2CH3), or -SON2N(CH2CH2OCH3)2.

[0278] In certain embodiments, each —R of formula (X) 4are independently C1-C3 alkyl. In certain embodiments, both -R 4 is methyl.

[0279] In certain embodiments, -Y- in formula (X) is absent. In certain embodiments, -Y- in formula (X) is -N(R 6 )CH2-.

[0280] In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is -CN and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is -SO2N(CH3)2 and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is SO2CH3, -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is -SO2N(CH2CH2)2CHCH3, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is phenyl substituted with -SO2, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is phenyl substituted with -SO2 and -Cl, -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1- is of formula (X), n is 1, and -R 1 is -SO2N(CH2CH2)2O, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is -SO2CH(CH3)2 and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is -SO2N(CH3)(CH2CH3), and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is -SO2N(CH2CH2OCH3)2, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 1, and -R 1 is phenyl substituted with -SO2 and -CH3, and -R 2 is -H and -R 4 is -CH3.

[0281] In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is -CN and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is -SO2N(CH3)2 and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is SO2CH3, -R 2 is -H and -R4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is -SO2N(CH2CH2)2CHCH3, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is phenyl substituted with -SO2, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is phenyl substituted with -SO2 and -Cl, -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is -SO2N(CH2CH2)2O, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is -SO2CH(CH3)2 and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is -SO2N(CH3)(CH2CH3), and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1 is -SO2N(CH2CH2OCH3)2, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 2, and -R 1is phenyl substituted with -SO2 and -CH3, and -R 2 is -H and -R 4 is -CH3.

[0282] In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is -CN and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is -SO2N(CH3)2 and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is SO2CH3, -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is -SO2N(CH2CH2)2CHCH3, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is phenyl substituted with -SO2, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is phenyl substituted with -SO2 and -Cl, -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is -SO2N(CH2CH2)2O, and -R 2 is -H and -R 4is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is -SO2CH(CH3)2 and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is -SO2N(CH3)(CH2CH3), and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is -SO2N(CH2CH2OCH3)2, and -R 2 is -H and -R 4 is -CH3. In certain embodiments, -L 1 - is of formula (X), n is 3, and -R 1 is phenyl substituted with -SO2 and -CH3, and -R 2 is -H and -R 4 is -CH3.

[0283] In the context of formula (X) only, the terms used have the following meanings:

[0284] The term "alkyl" refers to a straight-chain, branched, or cyclic saturated hydrocarbon group of 1 to 20, 1 to 12, 1 to 8, 1 to 6, or 1 to 4 carbon atoms. In certain embodiments, an alkyl is straight-chain or branched. Examples of straight-chain or branched alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and n-decyl. In certain embodiments, an alkyl is cyclic. Examples of cyclic alkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentadienyl, and cyclohexyl.

[0285] The term "alkoxy" refers to an alkyl group attached to an oxygen, including methoxy, ethoxy, isopropoxy, cyclopropoxy, and cyclobutoxy.

[0286] The term "alkenyl" refers to a non-aromatic unsaturated hydrocarbon having a carbon-carbon double bond and 2 to 20, 2 to 12, 2 to 8, 2 to 6, or 2 to 4 carbon atoms.

[0287] The term "alkynyl" refers to a non-aromatic unsaturated hydrocarbon having a carbon-carbon triple bond and 2 to 20, 2 to 12, 2 to 8, 2 to 6, or 2 to 4 carbon atoms.

[0288] The term "aryl" refers to an aromatic hydrocarbon group of 6 to 18 carbons, preferably 6 to 10 carbons, including groups such as phenyl, naphthyl, and anthracenyl. The term "heteroaryl" refers to an aromatic ring containing 3 to 15 carbons and at least one N, O, or S atom, preferably 3 to 7 carbons and at least one N, O, or S atom, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, and indenyl.

[0289] In certain embodiments, the alkenyl, alkynyl, aryl, or heteroaryl moiety may be attached to the remainder of the molecule through an alkyl linkage. In these situations, the substituent is referred to as alkenylalkyl, alkynylalkyl, arylalkyl, or heteroarylalkyl, indicating that an alkylene moiety is located between the alkenyl, alkynyl, aryl, or heteroaryl moiety and the molecule to which the alkenyl, alkynyl, aryl, or heteroaryl is attached.

[0290] The term "halogen" or "halo" refers to bromo, fluoro, chloro and iodo.

[0291] The term "heterocyclic ring" or "heterocyclyl" refers to a 3- to 15-membered aromatic or non-aromatic ring containing at least one N, O, or S atom. Examples include piperidinyl, piperazinyl, tetrahydropyranyl, pyrrolidine, and tetrahydrofuranyl, as well as the exemplary groups provided above for the term "heteroaryl." In certain embodiments, the heterocyclic ring or heterocyclyl is non-aromatic. In certain embodiments, the heterocyclic ring or heterocyclyl is aromatic.

[0292] The term "optionally substituted" refers to a group that may be unsubstituted or substituted with one or more (e.g., 1, 2, 3, 4, or 5) substituents, which may be the same or different. Examples of substituents include alkyl, alkenyl, alkynyl, halogen, -CN, -OR aa , -SR aa , -NR aa R bb , -NO2, -C=NH(OR aa ), -C(O)R aa , -OC(O)R aa , -C(O)OR aa , -C(O)NR aa R bb , -OC(O)NR aa R bb , -NR aa C(O)R bb , -NR aa C(O)OR bb , -S(O)R aa , -S(O)2R aa , -NR aa S(O)R bb , -C(O)NR aa S(O)R bb , -NR aa S(O)2R bb , -C(O)NR aa S(O)2R bb , -S(O)NR aa R bb , -S(O)NR aa R bb , -P(O)(OR aa )(OR bb), heterocyclyl, heteroaryl, or aryl, wherein alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl, and aryl are each independently selected from -R cc is optionally replaced by -R aa and -R bb are each independently -H, alkyl, alkenyl, alkynyl, heterocyclyl, heteroaryl, or aryl, or -R aa and -R bb together with the nitrogen atom to which they are attached form a heterocyclyl, which is optionally substituted by alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkoxy, or -CN; and each -R cc is independently alkyl, alkenyl, alkynyl, halogen, heterocyclyl, heteroaryl, aryl, -CN, or -NO2.

[0293] In certain embodiments, -L 1 - has a structure as disclosed in WO 2021 / 136808 A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 is a group represented by the formula (XI):

[0294] [ka] [In the formula, The dashed line indicates the attachment of -D to the nitrogen of a primary or secondary amine; v is selected from the group consisting of 0 or 1; -X 1 - is -C(R 8 )(R 8a )-, -N(R 9 )- and -O-; =X 2 is =O and =N(R 10 ) selected from the group consisting of -X 3 is selected from the group consisting of —O, —S, and —Se; each p is independently selected from the group consisting of 0 or 1, provided that at most one p is 0; -R 6 , -R 6a , -R 10 -H, -C(R 11 )(R 11a )(R 11b ) and -T; -R 9 is -C(R 11 )(R 11a )(R 11b ) and -T; -R 1 , -R 1a , -R 2 , -R 2a , -R 3 , -R 3a , -R 4 , -R 4a , -R 5 , -R 5a , -R 7 , -R 8 -R 8a , -R 11 , -R 11a and -R 11b -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, independently selected from the group consisting of C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl may be one or more -R 13 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 14 )-, -S(O)2N(R 14 )-, -S(O)N(R 14 )-, -S(O)2-, -S(O)-, -N(R 14 )S(O)2N(R 14a )-, -S-, -N(R 14 )-, -OC(OR 14 )(R 14a )-, -N(R 14 )C(O)N(R 14a )- and -OC(O)N(R 14 )-, -R 12 , -R 12a , -R 12b -H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl may be one or more -R 13 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 14 )-, -S(O)2N(R 14 )-, -S(O)N(R 14 )-, -S(O)2-, -S(O)-, -N(R 14 )S(O)2N(R14a )-, -S-, -N(R 14 )-, -OC(OR 14 )(R 14a )-, -N(R 14 )C(O)N(R 14a )- and -OC(O)N(R 14 )-, Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 13 and optionally independently substituted with -R 13 is halogen, -CN, oxo, -C(O)OR 15 , -OR 15 , -C(O)R 15 , -C(O)N(R 15 )(R 15a ), -S(O)2N(R 15 )(R 15a ), -S(O)N(R 15 )(R 15a ), -S(O)R 15 , -S(O)R 15 , -N(R 15 )S(O)2N(R 15a )(R 15b ), -SR 15 , -N(R 15 )(R 15a ), -NO2, -OC(O)R 15 , -N(R 15 )C(O)R 15a , -N(R 15 )S(O)2R 15a , -N(R 15 )S(O)R 15a , -N(R 15 )C(O)OR 15a , -N(R 15 )C(O)N(R 15a )(R 15b ), -OC(O)N(R 15 )(R 15a ) and C 1~6alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 14 , -R 14a , -R 15 , -R 15a and -R 15b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Optionally, Pair-R 1 / -R 1a , -R 2 / -R 2a , -R 3 / -R 3a , -R 4 / -R 4a , -R 5 / -R 5a or -R 8 / -R 8a One or more of the following, together with the atoms to which they are attached, form a C 3~10 forming a cycloalkyl, a 3- to 10-membered heterocyclyl, or an 8- to 11-membered heterobicyclyl; Optionally, Pair-R 1 / -R 2 , -R 1 / -R 8 , -R 1 / -R 9 , -R 2 / -R 9 or -R 2 / -R 10 one or more of, taken together with the atoms to which they are attached, form a ring -A-; -A- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; Optionally, Pair-R 3 / -R 6 , -R 4 / -R 6 , -R 5 / -R 6 , -R6 / -R 6a or -R 6 / -R 7 one or more of, together with the atoms to which they are attached, form a ring -A'-; -A'- is selected from the group consisting of 3- to 10-membered heterocyclyl and 8- to 11-membered heterobicyclyl. It is of -L 1 -Ha-L 2 - and optionally further substituted, with the proviso that the hydrogen marked with an asterisk in formula (XI) is not replaced by a substituent.

[0295] In certain embodiments, the dashed line in formula (XI) indicates the bond of -D to the nitrogen atom of the primary amine. In certain embodiments, the dashed line in formula (XI) indicates the bond of -D to the nitrogen atom of the secondary amine.

[0296] In certain embodiments, -X in formula (XI) 3 is —O. In certain embodiments, —X in formula (XI) 3 In certain embodiments, -X in formula (XI) is -S. 3 is -Se.

[0297] In certain embodiments, -R of formula (XI) 6 is —H. In certain embodiments, —R of formula (XI) 6 is -C(R 11 )(R 11a )(R 11b In certain embodiments, -R of formula (XI) 6 is -T.

[0298] In certain embodiments, -R of formula (XI) 6a is —H. In certain embodiments, —R of formula (XI) 6a is -C(R 11 )(R 11a )(R 11b In certain embodiments, -R of formula (XI)6a is -T.

[0299] In certain embodiments, -R of formula (XI) 6 and -R 6a Both are -H.

[0300] In certain embodiments, v in formula (XI) is 0. In certain embodiments, v in formula (XI) is 1.

[0301] In certain embodiments, -X in formula (XI) 1 - is -C(R 8 )(R 8a In certain embodiments, -X in formula (XI) 1 - is -N(R 9 In certain embodiments, -X in formula (XI) 1 - is -O-.

[0302] In certain embodiments, the =X of formula (XI) 2 is ═O. In certain embodiments, ═X in formula (XI) 2 is =N(R 10 )

[0303] In certain embodiments, -R of formula (XI) 9 is -C(R 11 )(R 11a )(R 11b In certain embodiments, -R of formula (XI) 9 is -T.

[0304] In certain embodiments, -R of formula (XI) 10 is —H. In certain embodiments, —R of formula (XI) 10 is -C(R 11 )(R 11a )(R 11b In certain embodiments, -R of formula (XI) 10 is -T.

[0305] In certain embodiments, -R of formula (XI)1 -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 1 is —H. In certain embodiments, —R of formula (XI) 1 is halogen. In certain embodiments, —R of formula (XI) 1 is -T. In certain embodiments, -R of formula (XI) 1 is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 1 is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 1 is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 1 is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0306] In certain embodiments, -R of formula (XI) 1a -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 1a is —H. In certain embodiments, —R of formula (XI) 1a is halogen. In certain embodiments, —R of formula (XI) 1a is -T. In certain embodiments, -R of formula (XI) 1a is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 1a is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 1a is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 1ais selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0307] In certain embodiments, -R of formula (XI) 2 -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 2 is —H. In certain embodiments, —R of formula (XI) 2 is halogen. In certain embodiments, —R of formula (XI) 2 is -T. In certain embodiments, -R of formula (XI) 2 is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 2 is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 2 is C2~6 In certain embodiments, -R of formula (XI) is alkynyl. 2 is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0308] In certain embodiments, -R of formula (XI) 2a -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 2a is —H. In certain embodiments, —R of formula (XI) 2a is halogen. In certain embodiments, —R of formula (XI) 2a is -T. In certain embodiments, -R of formula (XI) 2a is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 2a is C2~6 In certain embodiments, -R of formula (XI) is alkenyl. 2a is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 2a is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0309] In certain embodiments, -R of formula (XI) 3 -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 3 is —H. In certain embodiments, —R of formula (XI) 3 is halogen. In certain embodiments, —R of formula (XI) 3 is -T. In certain embodiments, -R of formula (XI) 3 is C1~6 In certain embodiments, -R of formula (XI) is alkyl. 3 is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 3 is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 3 is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0310] In certain embodiments, -R of formula (XI) 3a -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 3a is —H. In certain embodiments, —R of formula (XI) 3a is halogen. In certain embodiments, —R of formula (XI)3a is -T. In certain embodiments, -R of formula (XI) 3a is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 3a is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 3a is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 3a is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0311] In certain embodiments, -R of formula (XI) 4 -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 4is —H. In certain embodiments, —R of formula (XI) 4 is halogen. In certain embodiments, —R of formula (XI) 4 is -T. In certain embodiments, -R of formula (XI) 4 is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 4 is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 4 is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 4 is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0312] In certain embodiments, -R of formula (XI) 4a -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 4a is —H. In certain embodiments, —R of formula (XI) 4a is halogen. In certain embodiments, —R of formula (XI) 4a is -T. In certain embodiments, -R of formula (XI) 4a is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 4a is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 4a is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 4a is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0313] In certain embodiments, -R of formula (XI) 5 -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 5 is —H. In certain embodiments, —R of formula (XI) 5 is halogen. In certain embodiments, —R of formula (XI) 5 is -T. In certain embodiments, -R of formula (XI) 5 is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 5 is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 5 is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 5 is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0314] In certain embodiments, -R of formula (XI) 5a -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 5a is —H. In certain embodiments, —R of formula (XI) 5a is halogen. In certain embodiments, —R of formula (XI) 5a is -T. In certain embodiments, -R of formula (XI) 5a is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 5a is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 5a is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 5a is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0315] In certain embodiments, -R of formula (XI) 7 -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 7 is —H. In certain embodiments, —R of formula (XI) 7 is halogen. In certain embodiments, —R of formula (XI) 7 is -T. In certain embodiments, -R of formula (XI) 7 is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 7 is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 7 is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 7 is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0316] In certain embodiments, -R of formula (XI) 8 -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 8 is —H. In certain embodiments, —R of formula (XI) 8 is halogen. In certain embodiments, —R of formula (XI) 8 is -T. In certain embodiments, -R of formula (XI) 8 is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 8 is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 8 is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 8 is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0317] In certain embodiments, -R of formula (XI) 8a -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 8a is —H. In certain embodiments, —R of formula (XI) 8a is halogen. In certain embodiments, —R of formula (XI) 8a is -T. In certain embodiments, -R of formula (XI) 8a is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 8a is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 8a is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 8a is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0318] In certain embodiments, -R of formula (XI) 11 -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a ), -S(O)2N(R 12 )(R 12a ), -S(O)N(R12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 11 is —H. In certain embodiments, —R of formula (XI) 11 is halogen. In certain embodiments, —R of formula (XI) 11 is -T. In certain embodiments, -R of formula (XI) 11 is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 11 is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 11 is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 11 is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0319] In certain embodiments, -R of formula (XI) 11a -H, halogen, -CN, -C(O)OR 12 , -OR 12 , -C(O)R 12 , -C(O)N(R 12 )(R 12a), -S(O)2N(R 12 )(R 12a ), -S(O)N(R 12 )(R 12a ), -S(O)R 12 , -S(O)R 12 , -N(R 12 )S(O)2N(R 12a )(R 12b ), -SR 12 , -NO2, -N(R 12 )C(O)OR 12a , -N(R 12 )C(O)N(R 12a )(R 12b ), -OC(O)N(R 12 )(R 12a ), -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 11a is —H. In certain embodiments, —R of formula (XI) 11a is halogen. In certain embodiments, —R of formula (XI) 11a is -T. In certain embodiments, -R of formula (XI) 11a is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 11a is C 2~6 In certain embodiments, - in formula (XI) is C 2~6 In certain embodiments, -R of formula (XI) is alkynyl. 11a is selected from the group consisting of -H, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, and 1-ethylpropyl.

[0320] In certain embodiments, -R of formula (XI) 12 -H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 12 is —H. In certain embodiments, —R of formula (XI) 12 is -T. In certain embodiments, -R of formula (XI) 12 is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 12 is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 12 is C 2~6 It is alkynyl.

[0321] In certain embodiments, -R of formula (XI) 12a -H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 12a is —H. In certain embodiments, —R of formula (XI) 12a is -T. In certain embodiments, -R of formula (XI) 12a is C 1~6 In certain embodiments, -R of formula (XI) is alkyl. 12a is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 12a is C 2~6 It is alkynyl.

[0322] In certain embodiments, -R of formula (XI) 12b -H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R of formula (XI) is selected from the group consisting of alkynyl. 12b is —H. In certain embodiments, —R of formula (XI) 12b is -T. In certain embodiments, -R of formula (XI) 12b is C 1~6 In certain embodiments, -R of formula (XI) is alkyl.12b is C 2~6 In certain embodiments, -R of formula (XI) is alkenyl. 12b is C 2~6 It is alkynyl.

[0323] In certain embodiments, T in formula (XI) is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 In certain embodiments, T in formula (XI) is selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl. In certain embodiments, T in formula (XI) is phenyl. In certain embodiments, T in formula (XI) is naphthyl. In certain embodiments, T in formula (XI) is indenyl. In certain embodiments, T in formula (XI) is indanyl. In certain embodiments, T in formula (XI) is tetralinyl. In certain embodiments, T in formula (XI) is tetralinyl. In certain embodiments, T in formula (XI) is C 3~10 In certain embodiments, T of formula (XI) is 3-10 membered heterocyclyl. In certain embodiments, T of formula (XI) is 8-11 membered heterobicyclyl.

[0324] In certain embodiments, T of formula (XI) is one or more —R 13 is replaced by .

[0325] In certain embodiments, T of formula (XI) is one —R of formula (XI). 13 is replaced by .

[0326] In certain embodiments, T in formula (XI) is -R 13 is not replaced by

[0327] In certain embodiments, -R of formula (XI) 13 is halogen, -CN, oxo, -C(O)OR 15 , -OR 15 , -C(O)R 15 , -C(O)N(R 15 )(R15a ), -S(O)2N(R 15 )(R 15a ), -S(O)N(R 15 )(R 15a ), -S(O)R 15 , -S(O)R 15 , -N(R 15 )S(O)2N(R 15a )(R 15b ), -SR 15 , -N(R 15 )(R 15a ), -NO2, -OC(O)R 15 , -N(R 15 )C(O)R 15a , -N(R 15 )S(O)2R 15a , -N(R 15 )S(O)R 15a , -N(R 15 )C(O)OR 15a , -N(R 15 )C(O)N(R 15a )(R 15b ), -OC(O)N(R 15 )(R 15a ) and C 1~6 In certain embodiments, the -R of formula (XI) is selected from the group consisting of alkyl. 13 is halogen. In certain embodiments, —R of formula (XI) 13 In certain embodiments, -R of formula (XI) is -CN. 13 In certain embodiments, -R of formula (XI) is oxo. 13 is -C(O)OR 15 In certain embodiments, -R of formula (XI) 13 -OR 15 In certain embodiments, -R of formula (XI) 13 is -C(O)R 15 In certain embodiments, -R of formula (XI) 13 is -C(O)N(R 15 )(R 15a In certain embodiments, -R of formula (XI) 13 is -S(O)2N(R 15 )(R 15aIn certain embodiments, -R of formula (XI) 13 is -S(O)N(R 15 )(R 15a In certain embodiments, -R of formula (XI) 13 is -S(O)2R 15 In certain embodiments, -R of formula (XI) 13 is -S(O)R 15 In certain embodiments, -R of formula (XI) 13 is -N(R 15 )S(O)2N(R 15a )(R 15b In certain embodiments, -R of formula (XI) 13 -SR 15 In certain embodiments, -R of formula (XI) 13 is -N(R 15 )(R 15a In certain embodiments, -R of formula (XI) 13 In certain embodiments, -R of formula (XI) is -NO. 13 is -OC(O)R 15 In certain embodiments, -R of formula (XI) 13 is -N(R 15 )C(O)R 15a In certain embodiments, -R of formula (XI) 13 is -N(R 15 )S(O)2R 15a In certain embodiments, -R of formula (XI) 13 is -N(R 15 )S(O)R 15a In certain embodiments, -R of formula (XI) 13 is -N(R 15 )C(O)OR 15a In certain embodiments, -R of formula (XI) 13 is -N(R 15 )C(O)N(R 15a )(R 15b In certain embodiments, -R of formula (XI) 13 is -OC(O)N(R 15 )(R 15aIn certain embodiments, -R of formula (XI) 13 is C 1~6 It is alkyl.

[0328] In certain embodiments, -R of formula (XI) 14 -H and C 1~6 In certain embodiments, the -R of formula (XI) is selected from the group consisting of alkyl. 14 is —H. In certain embodiments, —R of formula (XI) 14 is C 1~6 It is alkyl.

[0329] In certain embodiments, -R of formula (XI) 14a -H and C 1~6 In certain embodiments, the -R of formula (XI) is selected from the group consisting of alkyl. 14a is —H. In certain embodiments, —R of formula (XI) 14a is C 1~6 It is alkyl.

[0330] In certain embodiments, -R of formula (XI) 15 -H and C 1~6 In certain embodiments, the -R of formula (XI) is selected from the group consisting of alkyl. 15 is —H. In certain embodiments, —R of formula (XI) 15 is C 1~6 It is alkyl.

[0331] In certain embodiments, -R of formula (XI) 15a -H and C 1~6 In certain embodiments, the -R of formula (XI) is selected from the group consisting of alkyl. 15a is —H. In certain embodiments, —R of formula (XI) 15a is C 1~6 It is alkyl.

[0332] In certain embodiments, -R of formula (XI) 15b -H and C 1~6In certain embodiments, the -R of formula (XI) is selected from the group consisting of alkyl. 15b is —H. In certain embodiments, —R of formula (XI) 15b is C 1~6 It is alkyl.

[0333] In certain embodiments, -R of formula (XI) 1 and -R 1a together with the atoms to which they are attached, C 3~10 In certain embodiments, -R of formula (XI) forms a cycloalkyl. 1 and -R 1a together with the atoms to which they are attached form a 3- to 10-membered heterocyclyl. In certain embodiments, -R in formula (XI) 1 and -R 1a together with the atoms to which they are attached form an 8- to 11-membered heterobicyclyl.

[0334] In certain embodiments, -R of formula (XI) 2 and -R 2a together with the atoms to which they are attached, C 3~10 In certain embodiments, -R of formula (XI) forms a cycloalkyl. 2 and -R 2a together with the atoms to which they are attached form a 3- to 10-membered heterocyclyl. In certain embodiments, -R in formula (XI) 2 and -R 2a together with the atoms to which they are attached form an 8- to 11-membered heterobicyclyl.

[0335] In certain embodiments, -R of formula (XI) 3 and -R 3a together with the atoms to which they are attached, C 3~10 In certain embodiments, -R of formula (XI) forms a cycloalkyl. 3 and -R 3a together with the atoms to which they are attached form a 3- to 10-membered heterocyclyl. In certain embodiments, -R in formula (XI)3 and -R 3a together with the atoms to which they are attached form an 8- to 11-membered heterobicyclyl.

[0336] In certain embodiments, -R of formula (XI) 4 and -R 4a together with the atoms to which they are attached, C 3~10 In certain embodiments, -R of formula (XI) forms a cycloalkyl. 4 and -R 4a together with the atoms to which they are attached form a 3- to 10-membered heterocyclyl. In certain embodiments, -R in formula (XI) 4 and -R 4a together with the atoms to which they are attached form an 8- to 11-membered heterobicyclyl.

[0337] In certain embodiments, -R of formula (XI) 5 and -R 5a together with the atoms to which they are attached, C 3~10 In certain embodiments, -R of formula (XI) forms a cycloalkyl. 5 and -R 5a together with the atoms to which they are attached form a 3- to 10-membered heterocyclyl. In certain embodiments, -R in formula (XI) 5 and -R 5a together with the atoms to which they are attached form an 8- to 11-membered heterobicyclyl.

[0338] In certain embodiments, -R of formula (XI) 8 and -R 8a together with the atoms to which they are attached, C 3~10 In certain embodiments, -R of formula (XI) forms a cycloalkyl. 8 and -R 8a together with the atoms to which they are attached form a 3- to 10-membered heterocyclyl. In certain embodiments, -R in formula (XI) 8 and -R 8atogether with the atoms to which they are attached form an 8- to 11-membered heterobicyclyl.

[0339] In certain embodiments, -R of formula (XI) 1 and -R 2 together with the atoms to which they are attached form a ring -A- of formula (XI).

[0340] In certain embodiments, -R of formula (XI) 1 and -R 8 together with the atoms to which they are attached form a ring -A- of formula (XI).

[0341] In certain embodiments, -R of formula (XI) 1 and -R 9 together with the atoms to which they are attached form a ring -A- of formula (XI).

[0342] In certain embodiments, -R of formula (XI) 2 and -R 9 together with the atoms to which they are attached form a ring -A- of formula (XI).

[0343] In certain embodiments, -R of formula (XI) 2 and -R 10 together with the atoms to which they are attached form a ring -A- of formula (XI).

[0344] In certain embodiments, -A- in formula (XI) is phenyl. In certain embodiments, -A- in formula (XI) is naphthyl. In certain embodiments, -A- in formula (XI) is indenyl. In certain embodiments, -A- in formula (XI) is indanyl. In certain embodiments, -A- in formula (XI) is tetralinyl. In certain embodiments, -A- in formula (XI) is C 3~10 In certain embodiments, -A- in formula (XI) is 3- to 10-membered heterocyclyl. In certain embodiments, -A- in formula (XI) is 8- to 11-membered heterobicyclyl.

[0345] In certain embodiments, -R of formula (XI) 3 and -R 6 together with the atoms to which they are attached form a ring -A'- of formula (XI).

[0346] In certain embodiments, -R of formula (XI) 4 and -R 6 together with the atoms to which they are attached form a ring -A'- of formula (XI).

[0347] In certain embodiments, -R of formula (XI) 5 and -R 6 together with the atoms to which they are attached form a ring -A'- of formula (XI).

[0348] In certain embodiments, -R of formula (XI) 6 and -R 6a together with the atoms to which they are attached form a ring -A'- of formula (XI).

[0349] In certain embodiments, -R of formula (XI) 6 and -R 7 together with the atoms to which they are attached form a ring -A'- of formula (XI).

[0350] In certain embodiments, -A'- in formula (XI) is 3-10 membered heterocyclyl. In certain embodiments, -A'- in formula (XI) is 8-11 membered heterobicyclyl.

[0351] In certain embodiments, -L 1 is a group of formula (XIa):

[0352] [ka] [In the formula, The dashed line indicates the attachment of -D to the nitrogen of a primary or secondary amine; -R1 , -R 1a , -R 2 , -R 2a , -R 3 , -R 3a , -R 5 , -R 5a , -R 6 and -R 6a is used as defined in formula (XI). It is of -L 1 -Ha-L 2 - and optionally further substituted, with the proviso that the hydrogen marked with an asterisk in formula (XIa) is not replaced by a substituent.

[0353] In certain embodiments, the dashed line in Formula (XIa) indicates the bond of -D to the nitrogen atom of the primary amine. In certain embodiments, the dashed line in Formula (XIa) indicates the bond of -D to the nitrogen atom of the secondary amine.

[0354] In certain embodiments, -R 1 is —H. In certain embodiments, —R 1a is —H. In certain embodiments, —R 2 is —H. In certain embodiments, —R 2a is —H. In certain embodiments, —R 3 is —H. In certain embodiments, —R 3a is —H. In certain embodiments, —R 5 is —H. In certain embodiments, —R 5a is —H. In certain embodiments, —R 6 is —H. In certain embodiments, —R 6a is -H.

[0355] In certain embodiments, -L of formula (XIa) 1 - is not further substituted.

[0356] In certain embodiments, -R 1is -H, and this -H is -L 2 In certain embodiments, -R 1a is -H, and this -H is -L 2 In certain embodiments, -R 2 is -H, and this -H is -L 2 In certain embodiments, -R 2a is -H, and this -H is -L 2 In certain embodiments, -R 3 is -H, and this -H is -L 2 In certain embodiments, -R 3a is -H, and this -H is -L 2 In certain embodiments, -R 5 is -H, and this -H is -L 2 In certain embodiments, -R 5a is -H, and this -H is -L 2 In certain embodiments, -R 6 is -H, and this -H is -L 2 In certain embodiments, -R 6a is -H, and this -H is -L 2 - is replaced by

[0357] In certain embodiments, -L 1 is a group of formula (XIb):

[0358] [ka] [In the formula, The dashed line indicates the attachment of -D to the nitrogen of a primary or secondary amine. It is of -L 1 -Ha-L 2 - and optionally further substituted, with the proviso that the hydrogen marked with an asterisk in formula (XIb) is not replaced by a substituent.

[0359] In certain embodiments, the dashed line in Formula (XIb) indicates the bond of -D to the nitrogen atom of the primary amine. In certain embodiments, the dashed line in Formula (XIb) indicates the bond of -D to the nitrogen atom of the secondary amine.

[0360] In certain embodiments, -L of formula (XIb) 1 - is not further substituted.

[0361] In certain embodiments, -L 1 - is a group represented by the formula (XIc):

[0362] [ka] [In the formula, The unmarked dashed line indicates the attachment of -D to the nitrogen of a primary or secondary amine; The dashed line with # means -L 2 - indicates a bond to It is of the type.

[0363] In certain embodiments, the unmarked dashed line in Formula (XIc) indicates the bond of -D to the nitrogen atom of the primary amine. In certain embodiments, the unmarked dashed line in Formula (XIc) indicates the bond of -D to the nitrogen atom of the secondary amine.

[0364] In certain embodiments, -L 1 - has a structure as disclosed in WO 2020 / 254603 A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 - is a group represented by the formula (XII):

[0365] [ka] [In the formula, The dashed line indicates the bond of -D to the π electron pair donating heteroaromatic N; n is an integer selected from the group consisting of 0, 1, 2, 3, and 4; =X 1 are =O, =S and =N(R 4 ) selected from the group consisting of -X 2 - is -O-, -S-, -N(R 5 )- and -C(R 6 )(R 6a )-, -X 3 -teeth,

[0366] [ka] -C(R 10 )(R 10a )-, -C(R 11 )(R 11a )-C(R 12 )(R 12a )—, —O—, and —C(O)—; -R 1 , -R 1a , -R 6 , -R 6a , -R 10 , -R 10a , -R 11 , -R 11a , -R 12 , -R 12a , and -R 2 and -R 2a are -H, -C(O)OH, halogen, -CN, -OH, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, independently selected from the group consisting of C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl may be one or more -R 13 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 14 )-, -S(O)2N(R 14 )-, -S(O)N(R 14)-, -S(O)2-, -S(O)-, -N(R 14 )S(O)2N(R 14a )-, -S-, -N(R 14 )-, -OC(OR 14 )(R 14a )-, -N(R 14 )C(O)N(R 14a )- and -OC(O)N(R 14 )-, -R 3 , -R 4 , -R 5 , -R 7 , -R 8 and -R 9 -H, -T, -CN, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, independently selected from the group consisting of C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl may be one or more -R 13 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 14 )-, -S(O)2N(R 14 )-, -S(O)N(R 14 )-, -S(O)2-, -S(O)-, -N(R 14 )S(O)2N(R 14a )-, -S-, -N(R 14 )-, -OC(OR 14 )(R 14a )-, -N(R 14 )C(O)N(R 14a )- and -OC(O)N(R 14 )-, Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 13 and optionally independently substituted with -R 13 -H, -NO2, -OCH3, -CN, -N(R 14 )(R 14a ), -OH, -C(O)OH and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 14 and -R 14a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Optionally, Pair-R 1 / -R 1a , -R 2 / -R 2a , two adjacent -R 2 , -R 6 / -R 6a , -R 10 / -R 10a , -R 11 / -R 11a and -R 12 / -R 12a One or more of the following, together with the atoms to which they are attached, form a C 3~10 forming a cycloalkyl, a 3- to 10-membered heterocyclyl, or an 8- to 11-membered heterobicyclyl; Optionally, Pair-R 1 / -R 2 , -R 1 / -R 5 , -R 1 / -R 6 , -R 1 / -R 9 , -R 1 / -R 10 , -R 3 / -R 6a , -R 4 / -R 5 , -R 4 / -R6 , -R 5 / -R 10 and -R 6 / -R 10 one or more of, taken together with the atoms to which they are attached, form a ring -A-; -A- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; Optionally, -R 1 and adjacent -R 2 forms a carbon-carbon double bond, with the proviso that n is selected from the group consisting of 1, 2, 3 and 4; Optionally, two adjacent -R 2 forms a carbon-carbon double bond, with the proviso that n is selected from the group consisting of 2, 3, and 4; However, -X 2 - is -N(R 5 )-, then -X 3 -teeth,

[0367] [ka] and the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk in formula (XII) is 5, 6 or 7 atoms, and, if present, -R 1 and -R 2 Between or between two adjacent -R 2 and the carbon-carbon double bond formed between them is in a cis configuration. It is of Each-L 1 -Ha-L 2 - and optionally further substituted.

[0368] It is understood that the "N" in the phrase "π electron pair donating heteroaromatic N" refers to nitrogen.

[0369] Two adjacent -R in formula (XII) 2It is understood that can only be present if n is at least 2.

[0370] The phrase "the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk" refers to the total number of atoms in the shortest distance between the nitrogen atom marked with an asterisk and the carbon atom, and is understood to include the nitrogen atom marked with an asterisk and the carbon atom. For example, in the following structure, n is 1 and the distance between the nitrogen atom marked with an asterisk and the carbon atom is 5:

[0371] [ka] In the following structure, n is 2 and -R 1 and -R 1a forms a cyclohexyl, and the distance between the starred nitrogen and the starred carbon is 6:

[0372] [ka]

[0373] In certain embodiments, the =X of formula (XII) 1 is ═O. In certain embodiments, ═X in formula (XII) 1 is =S. In certain embodiments, =X in formula (XII) 1 is =N(R 4 )

[0374] In certain embodiments, -X in formula (XII) 2 In certain embodiments, -X in formula (XII) 2 - is -S-. In certain embodiments, -X in formula (XII) 2 - is -N(R 5 In certain embodiments, -X in formula (XII) is 2 - is -C(R 6 )(R 6a )-.

[0375] In certain embodiments, -X in formula (XII)3 -teeth,

[0376] [ka] is.

[0377] In certain embodiments, -X in formula (XII) 3 -teeth,

[0378] [ka] is.

[0379] In certain embodiments, -X in formula (XII) 3 -teeth,

[0380] [ka] is.

[0381] In certain embodiments, -X in formula (XII) 3 - is -C(R 10 )(R 10a In certain embodiments, -X in formula (XII) is 3 - is -C(R 11 )(R 11a )-C(R 12 )(R 12a In certain embodiments, -X in formula (XII) is 3 In certain embodiments, -X in formula (XII) 3 - is -C(O)-.

[0382] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0383] [ka] and the distance between the starred nitrogen atom and the starred carbon atom in formula (XII) is 5 atoms.

[0384] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0385] [ka] and the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk in formula (XII) is 6 atoms.

[0386] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0387] [ka] and the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk in formula (XII) is 7 atoms.

[0388] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0389] [ka] and the distance between the starred nitrogen atom and the starred carbon atom in formula (XII) is 5 atoms.

[0390] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0391] [ka] and the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk in formula (XII) is 6 atoms.

[0392] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0393] [ka] and the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk in formula (XII) is 7 atoms.

[0394] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0395] [ka] and the distance between the starred nitrogen atom and the starred carbon atom in formula (XII) is 5 atoms.

[0396] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0397] [ka] and the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk in formula (XII) is 6 atoms.

[0398] In certain embodiments, -X in formula (XII) 2 -is-N(R5 )-, and -X in formula (XII) 3 -teeth,

[0399] [ka] and the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk in formula (XII) is 7 atoms.

[0400] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0401] [ka] and the distance between the starred nitrogen atom and the starred carbon atom in formula (XII) is 5 atoms.

[0402] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0403] [ka] and the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk in formula (XII) is 6 atoms.

[0404] In certain embodiments, -X in formula (XII) 2 -is-N(R 5 )-, and -X in formula (XII) 3 -teeth,

[0405] [ka] and the distance between the nitrogen atom marked with an asterisk and the carbon atom marked with an asterisk in formula (XII) is 7 atoms.

[0406] In certain embodiments, the =X of formula (XII) 1 is ═O, and -X in formula (XII) 2 -is-C(R 6 )(R 6a )-, and -X in formula (XII) 3 -teeth,

[0407] [ka] and -R 3 does not contain amines.

[0408] In certain embodiments, -R of formula (XII) 1 , -R 1a , -R 6 , -R 6a , -R 10 , -R 10a , -R 11 , -R 11a , -R 12 , -R 12a and -R of formula (XII) 2 and -R 2a are -H, -C(O)OH, halogen, -CN, -OH, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl.

[0409] In certain embodiments, -L 1 - has a structure as disclosed in WO 2020 / 254602 A1, which is incorporated herein by reference in its entirety. 1 -L 1 - when bonded to an electron-donating heteroaromatic N + moiety or a quaternary ammonium cation and when linked, the moiety -D + Thus, in certain embodiments, -L 1 - is a group represented by the formula (XII):

[0410] [ka] [In the formula, The dashed line is -D + N + indicates a bond to t is selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6; -A- is a monocyclic or bicyclic ring selected from the group consisting of aryl and heteroaryl, provided that -A- is connected via a carbon atom to -Y and -C(R 1 )(R 1a )-, provided that the monocyclic or bicyclic aryl and heteroaryl are connected to one or more -R 2 is optionally replaced by -R 1 , -R 1a and each -R 2 are -H, -C(O)OH, -halogen, -NO2, -CN, -OH, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, independently selected from the group consisting of C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl may be one or more -R 3 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 4 )-, -S(O)2N(R 4 )-, -S(O)N(R 4 )-, -S(O)2-, -S(O)-, -N(R 4 )S(O)2N(R 4a )-, -S-, -N(R 4 )-, -OC(OR 4 )(R 4a )-, -N(R 4 )C(O)N(R 4a )- and -OC(O)N(R 4 )-, Each -T- is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; each -T- is the same or different, and is selected from the group consisting of one or more -R 3 and optionally independently substituted with -R 3 -H, -NO2, -OCH3, -CN, -N(R 4 )(R 4a ), -OH, -C(O)OH and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 4 and -R 4a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -Y is

[0411] [ka] (In the formula, The dashed line with an asterisk indicates the bond to -A-; -Nu is a nucleophile, -Y 1 - is -O-, -C(R 10 )(R 10a )-, -N(R 11 )- and -S-; =Y 2 are =O, =S and =N(R 12 ) selected from the group consisting of -Y 3 - is -O-, -S- and -N(R 13 )-, -E- is C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 alkynyl and -Q-; 1~6 Alkyl, C2~6 Alkenyl, C 2~6 Alkynyl may be one or more -R 14 is optionally replaced by -R 5 , -R 6 , each -R 7 , -R 8 , -R 9 , -R 10 , -R 10a , -R 11 , -R 12 and -R 13 is C 1~20 Alkyl, C 2~20 Alkenyl, C 2~20 independently selected from the group consisting of alkynyl and -Q; 1~20 Alkyl, C 2~20 Alkenyl and C 2~20 Alkynyl may be one or more -R 14 is optionally replaced by C 1~20 Alkyl, C 2~20 Alkenyl and C 2~20 Alkynyl is -Q-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 15 )-, -S(O)2N(R 15 )-, -S(O)N(R 15 )-, -S(O)2-, -S(O)-, -N(R 15 )S(O)2N(R 15a )-, -S-, -N(R 15 )-, -OC(OR 15 )R 15a -, -N(R 15 )C(O)N(R 15a )- and -OC(O)N(R 15 )-, Each Q is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; each Q is the same or different and is selected from the group consisting of one or more -R 14 and optionally independently substituted with -R 14 , -R15 and -R 15a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. selected from the group consisting of It is of Each-L 1 -Ha-L 2 - and optionally further substituted.

[0412] In certain embodiments, -D + is an electron-donating heteroaromatic N + It is understood that D may include both an electron donating heteroaromatic N- and a quaternary ammonium cation, and similarly, the corresponding D may include both an electron donating heteroaromatic N- and a tertiary amine. 1 -D when conjugated to - + and -L 1 It is also understood that - forms a quaternary ammonium cation, for which a counteranion may be present. Examples of counteranions include, but are not limited to, chloride, bromide, acetate, bicarbonate, sulfate, bisulfate, nitrate, carbonate, alkylsulfonate, arylsulfonate, and phosphate.

[0413] -L of formula (XIII) 1 The optional further substituents of - are as described elsewhere herein. In certain embodiments, -L of formula (XIII) 1 - is not further substituted.

[0414] Such drug moieties -D + is at least one, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 electron-donating heteroaromatic N +or a quaternary ammonium cation, and similarly, the corresponding released drug D contains at least one, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, electron-donating heteroaromatic N or tertiary amine. The heteroaromatic nitrogen atom that donates electrons to the aromatic π system, i.e., N + Examples of chemical structures containing N include, but are not limited to, pyridine, pyridazine, pyrimidine, quinoline, quinazoline, quinoxaline, pyrazole, imidazole, isoindazole, indazole, purine, tetrazole, triazole, and triazine. For example, in the imidazole ring below, the heteroaromatic nitrogen that donates one electron to the aromatic pi-system is marked with a "§":

[0415] [ka]

[0416] Such electron-donating heteroaromatic nitrogen atoms do not include heteroaromatic nitrogen atoms that donate one electron pair (i.e., not one electron) to the aromatic π-system, such as the nitrogen marked with a "#" in the imidazole ring structure described above. Drug D may exist in one or more tautomeric forms, for example, in which one hydrogen atom migrates between at least two heteroaromatic nitrogen atoms. In all such cases, the linker moiety is covalently and reversibly attached at the heteroaromatic nitrogen that donates an electron to the aromatic π-system.

[0417] As used herein, the term "monocyclic or bicyclic aryl" means an aromatic hydrocarbon ring system that may be monocyclic or bicyclic, where a monocyclic aryl ring consists of at least 5 ring carbon atoms and may contain up to 10 ring carbon atoms, and a bicyclic aryl ring consists of at least 8 ring carbon atoms and may contain up to 12 ring carbon atoms. Each hydrogen atom of a monocyclic or bicyclic aryl may be replaced by a substituent as defined below.

[0418] As used herein, the term "monocyclic or bicyclic heteroaryl" refers to a monocyclic aromatic ring system which may contain 2 to 6 ring carbon atoms and 1 to 3 ring heteroatoms, or a bicyclic aromatic ring system which may contain 3 to 9 ring carbon atoms and 1 to 5 ring heteroatoms, such as nitrogen, oxygen, and sulfur. Examples of monocyclic or bicyclic heteroaryl groups include, but are not limited to, benzofuranyl, benzothiophenyl, furanyl, imidazolyl, indolyl, azaindolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, tetrazinyl, tetrazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, quinolinyl, quinazolinyl, quinoxalinyl, triazolyl, thiazolyl, and thiophenyl. Each hydrogen atom of a monocyclic or bicyclic heteroaryl may be replaced by a substituent as defined below.

[0419] As used herein, the term "nucleophile" refers to a reagent or functional group that forms a bond with a reaction partner, i.e., an electrophile, by donating electrons to bond the two together.

[0420] In certain embodiments, t in Formula (XIII) is 0. In certain embodiments, t in Formula (XIII) is 1. In certain embodiments, t in Formula (XIII) is 2. In certain embodiments, t in Formula (XIII) is 3. In certain embodiments, t in Formula (XIII) is 4. In certain embodiments, t in Formula (XIII) is 5. In certain embodiments, t in Formula (XIII) is 6.

[0421] In certain embodiments, -A- in formula (XIII) is a ring selected from the group consisting of monocyclic or bicyclic aryl and heteroaryl, provided that -A- is connected through a carbon atom to -Y and -C(R 1 )(R 1a In certain embodiments, -A- of formula (XIII) is connected to one or more -R of formula (XIII), which may be the same or different.2 In certain embodiments, -A- of formula (XIII) is substituted with -R of formula (XIII). 2 In certain embodiments, -A- in formula (XIII) is not substituted with

[0422] [ka] wherein each V is independently selected from the group consisting of O, S, and N. is selected from the group consisting of:

[0423] In certain embodiments, -R of formula (XIII) 1 , -R 1a and each -R 2 are -H, -C(O)OH, -halogen, -CN, -NO2, -OH, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R in formula (XIII) is independently selected from the group consisting of alkynyl. 1 , -R 1a and each -R 2 -H, -C(O)OH, -CN, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R in formula (XIII) is independently selected from the group consisting of alkynyl. 1 is —H. In certain embodiments, —R of formula (XIII) 1 is —C(O)OH. In certain embodiments, —R of formula (XIII) 1 In certain embodiments, -R of formula (XIII) is -halogen. 1 is -F. In certain embodiments, -R of formula (XIII) 1 In certain embodiments, -R in formula (XIII) is -CN. 1 In certain embodiments, -R of formula (XIII) is -NO. 1 In certain embodiments, -R of formula (XIII) is -OH. 1 is C 1~6In certain embodiments, -R of formula (XIII) is alkyl. 1 is C 2~6 In certain embodiments, -R of formula (XIII) is alkenyl. 1 is C 2~6 In certain embodiments, -R of formula (XIII) is alkynyl. 1a is —H. In certain embodiments, —R of formula (XIII) 1a is —C(O)OH. In certain embodiments, —R of formula (XIII) 1a In certain embodiments, -R of formula (XIII) is -halogen. 1a is -F. In certain embodiments, -R of formula (XIII) 1a In certain embodiments, -R in formula (XIII) is -CN. 1a In certain embodiments, -R of formula (XIII) is -NO. 1a In certain embodiments, -R of formula (XIII) is -OH. 1a is C 1~6 In certain embodiments, -R of formula (XIII) is alkyl. 1a is C 2~6 In certain embodiments, -R of formula (XIII) is alkenyl. 1a is C 2~6 It is alkynyl.

[0424] In certain embodiments, -R of formula (XIII) 2 are -H, -C(O)OH, -halogen, -CN, -NO2, -OH, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 In certain embodiments, -R in formula (XIII) is independently selected from the group consisting of alkynyl. 2 Each of -R in formula (XIII) is -H. 2 Each of -R in formula (XIII) is -C(O)OH. 2 In certain embodiments, each of -R in formula (XIII) is -halogen. 2Each of -R in formula (XIII) is -F. 2 In certain embodiments, each of -R in formula (XIII) is -CN. 2 In certain embodiments, each of -R in formula (XIII) is -NO. 2 In certain embodiments, each of -R in formula (XIII) is -OH. 2 Each of the 1~6 In certain embodiments, -R of formula (XIII) is alkyl. 2 Each of the 2~6 In certain embodiments, -R of formula (XIII) is alkenyl. 2 Each of the 2~6 It is alkynyl.

[0425] In certain embodiments, T in formula (XIII) is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 In certain embodiments, T in formula (XIII) is selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl. In certain embodiments, T in formula (XIII) is phenyl. In certain embodiments, T in formula (XIII) is naphthyl. In certain embodiments, T in formula (XIII) is indenyl. In certain embodiments, T in formula (XIII) is indanyl. In certain embodiments, T in formula (XIII) is tetralinyl. In certain embodiments, T in formula (XIII) is C 3~10 In certain embodiments, T of formula (XIII) is 3-10 membered heterocyclyl. In certain embodiments, T of formula (XIII) is 8-11 membered heterobicyclyl.

[0426] In certain embodiments, T of formula (XIII) is one or more —R 3 In certain embodiments, T of formula (XIII) is substituted with one -R of formula (XIII). 3 In certain embodiments, T of formula (XIII) is substituted with -R of formula (XIII).3 is not replaced by

[0427] In certain embodiments, -R of formula (XIII) 3 -H, -NO2, -OCH3, -CN, -N(R 4 )(R 4a ), -OH, -C(O)OH and C 1~6 In certain embodiments, the -R of formula (XIII) is selected from the group consisting of alkyl. 3 is —H. In certain embodiments, —R of formula (XIII) 3 In certain embodiments, -R of formula (XIII) is -NO. 3 In certain embodiments, -R in formula (XIII) is -OCH. 3 In certain embodiments, -R in formula (XIII) is -CN. 3 is -N(R 4 )(R 4a In certain embodiments, -R of formula (XIII) 3 In certain embodiments, -R of formula (XII) is -OH. 3 is —C(O)OH. In certain embodiments, —R of formula (XIII) 3 is C 1~6 In certain embodiments, -R of formula (XIII) is alkyl. 4 and -R 4a -H and C 1~6 In certain embodiments, -R of formula (XIII) is independently selected from the group consisting of alkyl. 4 is -H. In certain embodiments, -R 4 is C 1~6 In certain embodiments, -R of formula (XIII) is alkyl. 4a is —H. In certain embodiments, —R of formula (XIII) 4a is C 1~6 It is alkyl.

[0428] In certain embodiments, -Y in formula (XIII) is

[0429] [ka] TIFF2026501742000073.tif24163[where, -Nu, -E-, -Y 1 -, =Y 2 , -Y 3 -, -R 5 , -R 7 , -R 8 and -R 9 is defined as above] is selected from the group consisting of:

[0430] In certain embodiments, -L 1 - has a structure as disclosed in WO 2020 / 254606 A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the moiety -L 1 is a group represented by the formula (XIV):

[0431] [ka] [In the formula, The dashed line with the star indicates -L 2 - indicates a bond to The unmarked dashed line indicates the bond of -D to the π-electron pair donating heteroaromatic N; -Y- is -N(R 3 )—, —O—, and —S—; -R 1 , -R 2 and -R 3 -H, -T, C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 alkynyl, independently selected from the group consisting of C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6 Alkynyl may be one or more -R 4 is optionally replaced by C 1~6 Alkyl, C 2~6 Alkenyl and C 2~6Alkynyl is -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R 5 )-, -S(O)2N(R 5 )-, -S(O)N(R 5 )-, -S(O)2-, -S(O)-, -N(R 5 )S(O)2N(R 5a )-, -S-, -N(R 5 )-, -OC(OR 5 )(R 5a )-, -N(R 5 )C(O)N(R 5a )- and -OC(O)N(R 5 )-, Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 4 and optionally independently substituted with -R 4 , -R 5 and -R 5a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. It is of Each-L 1 -Ha-L 2 - and optionally further substituted.

[0432] In certain embodiments, -L 2 - does not exist.

[0433] In certain embodiments, -L 2 - is a spacer portion.

[0434] In certain embodiments, -L 2 -, -L 2' -or-L 2''- stands for -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O)2N(R y1 )-, -S(O)N(R y1 )-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-, -N(R y1 )C(O)N(R y1a )-, -OC(O)N(R y1 )-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl, -T'-, C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl may be one or more -R y2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-, -S(O)2N(R y3 )-, -S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-, -N(R y3 )C(O)N(R y3a )- and -OC(O)N(R y3 )-, -R y1 and -R y1a -H, -T', C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 alkynyl, -T', C 1~50 Alkyl, C2~50 Alkenyl and C 2~50 Alkynyl may be one or more -R y2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl and C 2~50 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 )-, -S(O)2N(R y4 )-, -S(O)N(R y4 )-, -S(O)2-, -S(O)-, -N(R y4 )S(O)2N(R y4a )-, -S-, -N(R y4 )-, -OC(OR y4 )(R y4a )-, -N(R y4 )C(O)N(R y4a )- and -OC(O)N(R y4 )-, Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; each T' is the same or different and is selected from the group consisting of one or more -R y2 and optionally independently substituted with Each-R y2 is halogen, -CN, oxo(=O), -C(O)OR y5 , -OR y5 , -C(O)R y5 , -C(O)N(R y5 )(R y5a ), -S(O)2N(R y5 )(R y5a ), -S(O)N(R y5 )(R y5a ), -S(O)R y5 , -S(O)R y5 , -N(R y5 )S(O)2N(R y5 )(R y5a ), -SR y5 , -N(Ry5 )(R y5a ), -NO2, -OC(O)R y5 , -N(R y5 )C(O)R y5a , -N(R y5 )S(O)2R y5a , -N(R y5 )S(O)R y5a , -N(R y5 )C(O)OR y5a , -N(R y5 )C(O)N(R y5 )(R y5a ), -OC(O)N(R y5 )(R y5a ), and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R y3 , -R y3a , -R y4 , -R y4a , -R y5 , -R y5a and -R y5b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different.

[0435] In certain embodiments, -L 2 -, -L 2' -or-L 2'' - stands for -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O)2N(R y1 )-, -S(O)N(R y1 )-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-, -N(R y1 )C(O)N(R y1a )-, -OC(O)N(R y1 )-, C 1~50Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl, -T'-, C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 Alkynyl may be one or more -R y2 is optionally replaced by C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-, -S(O)2N(R y3 )-, -S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-, -N(R y3 )C(O)N(R y3a )- and -OC(O)N(R y3 )-, -R y1 and -R y1a -H, -T', C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl, -T', C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 Alkynyl may be one or more -R y2 is optionally replaced by C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 )-, -S(O)2N(R y4 )-, -S(O)N(R y4 )-, -S(O)2-, -S(O)-, -N(R y4 )S(O)2N(R y4a)-, -S-, -N(R y4 )-, -OC(OR y4 )(R y4a )-, -N(R y4 )C(O)N(R y4a )- and -OC(O)N(R y4 )-, Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; each T' is the same or different and is selected from the group consisting of one or more -R y2 and optionally independently substituted with -R y2 is halogen, -CN, oxo(=O), -C(O)OR y5 , -OR y5 , -C(O)R y5 , -C(O)N(R y5 )(R y5a ), -S(O)2N(R y5 )(R y5a ), -S(O)N(R y5 )(R y5a ), -S(O)R y5 , -S(O)R y5 , -N(R y5 )S(O)2N(R y5a )(R y5b ), -SR y5 , -N(R y5 )(R y5a ), -NO2, -OC(O)R y5 , -N(R y5 )C(O)R y5a , -N(R y5 )S(O)2R y5a , -N(R y5 )S(O)R y5a , -N(R y5 )C(O)OR y5a , -N(R y5 )C(O)N(R y5a )(R y5b ), -OC(O)N(R y5 )(R y5a) and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R y3 , -R y3a , -R y4 , -R y4a , -R y5 , -R y5a and -R y5b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different.

[0436] In certain embodiments, -L 2 -, -L 2' -or-L 2'' - stands for -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O)2N(R y1 )-, -S(O)N(R y1 )-, -S(O)2-, -S(O)-, -N(R y1 )S(O)2N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-, -N(R y1 )C(O)N(R y1a )-, -OC(O)N(R y1 )-, C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl, -T'-, C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R y2 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-, -S(O)2N(R y3)-, -S(O)N(R y3 )-, -S(O)2-, -S(O)-, -N(R y3 )S(O)2N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-, -N(R y3 )C(O)N(R y3a )- and -OC(O)N(R y3 )-, -R y1 and -R y1a -H, -T', C 1~10 Alkyl, C 2~10 Alkenyl and C 2~10 alkynyl; Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 independently selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; Each-R y2 is a halogen, and C 1~6 alkyl, and each -R y3 , -R y3a , -R y4 , -R y4a , -R y5 , -R y5a and -R y5b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different.

[0437] In certain embodiments, -L 2 -, -L 2' -or-L 2'' -C 1~20 alkyl chains, which are -O-, -T'- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from the group consisting of1~20 The alkyl chains are -OH, -T' and -C(O)N(R y6 R y6a ) optionally substituted with one or more groups independently selected from the group consisting of -R y1 , -R y6 , -R y6a is H and C 1~4 alkyl, and T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 It is selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl.

[0438] In certain embodiments, -L 2 -, -L 2' -or-L 2'' - has a molecular weight in the range of 14 g / mol to 750 g / mol.

[0439] In certain embodiments, -L 2 -, -L 2' -or-L 2'' -teeth,

[0440] [ka] [In the formula, -L 2 - Regarding the dashed line, -L 1 - and -L 4 - indicates a bond to -R and -R a are independently selected from the group consisting of -H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, and 3,3-dimethylpropyl. The moiety comprises a moiety selected from the group consisting of:

[0441] In certain embodiments, -L 2- is the following part

[0442] [ka] Includes.

[0443] In certain embodiments, -L 2 -, -L 2' -or-L 2'' - has a chain length of 1 to 20 atoms.

[0444] Preferably, -L 2 - is the formula (l1):

[0445] [ka] [In the formula, the unmarked dashed line represents -L 4 - indicates a bond to -, while a dashed line with an asterisk indicates a bond to -L 1 - indicates a bond to It is of the type.

[0446] Preferably, -L 2 -L 1 - is the formula (s1):

[0447] [ka] [Wherein the formula, the dashed line represents -L 4 - indicates the bond to -, a dashed line with an asterisk indicates the bond to -D, -L 4 - and -D are defined as elsewhere herein. It is of the type.

[0448] Preferably, -L 2 -L 1 - is the formula (s2):

[0449] [ka] [Wherein the formula, the dashed line represents -L 4- indicates the bond to -, a dashed line with an asterisk indicates the bond to -D, -L 4 - and -D are defined as elsewhere herein. It is of the type.

[0450] Preferably, the attachment of -D to (s1) or (s2) occurs by forming an amide bond.

[0451] In certain embodiments, each -L 3 -, -L 4 -or-L 5 -teeth,

[0452] [ka] TIFF2026501742000081.tif222163TIFF2026501742000082.tif222163TIFF2026501742000083.tif222163TI FF2026501742000084.tif210164TIFF2026501742000085.tif224163TIFF2026501742000086.tif61164[In the expression, -L 3 For -, one dashed line indicates the bond to -X'- and the other dashed line indicates the bond to -Y'-, and -L 4 -, one dashed line is -L 2 The other dashed line represents the bond to -Y'-, and -L 5 For -, one dashed line indicates the bond to -Y'- and the other dashed line indicates the bond to -BA; -Y-, -O-, -S-, -NR 05 -, -CR 05 R 05a - selected from the group consisting of Each-R 04 , -R 04a , -R 04b , -R 04c , -R 05 and -R 05a are halogens, -H, -CN, -T 0 , C 1~50Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 06 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R0 7 )-, -S(O)2N(R 07 )-, -S(O)N(R 07 )-, -S(O)2-, -S(O)-, -N(R 07 )S(O)2N(R 07a )-, -S-, -N(R 07 )-, -OC(OR 07 )(R 07a )-, -N(R 07 )C(O)N(R 07a )- and -OC(O)N(R 07 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 06 and optionally independently substituted with Each-R 06 , -R 07 and -R 07a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. are independently selected from the group consisting of:

[0453] In certain embodiments, all moieties -L 3 In certain embodiments, all moieties -L 4 In certain embodiments, all moieties -L 5 In certain embodiments, all moieties -L 3 - has the same structure and all moieties -L 4 - has the same structure and all moieties -L 5 - have the same structure, which may be the same or different between different moieties. In certain embodiments, all moieties -L 3 -, All Parts-L 4 - and all parts - L 5 - have the same chemical structure.

[0454] Exemplary blocking agents include:

[0455] [ka] [Wherein the formula, the dashed line represents -L 5 - indicates a bond to may be selected from the group consisting of:

[0456] In certain embodiments, the blocking agent is of formula (b01) or (b02). In certain embodiments, the blocking agent is of formula (b01). In certain embodiments, the blocking agent is of formula (b02). In certain embodiments, the blocking agent is of formula (b03).

[0457] In certain embodiments, the drug conjugate or pharmaceutically acceptable salt thereof comprises an HA hydrogel microsphere comprising crosslinked HA chains to which a plurality of drug moieties are covalently and reversibly conjugated, or a pharmaceutically acceptable salt thereof, wherein the drug conjugate comprises each of the following units:

[0458] [ka] TIFF2026501742000089.tif85165[in the formula, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each R a1 , -R a2 , -X'-, -Y'-, -D, -L 1 -, -L 2 - are used as defined elsewhere herein] Contains multiple The drug conjugate has a Z in the range of about 50% to about 98% 1 , Z in the range of about 0.1% to about 20% 2 -i, Z in the range of about 0.1% to about 20% 3 -i and Z in the range of about 0.1% to about 10% 4 Includes -i.

[0459] In certain embodiments, the drug conjugate or pharmaceutically acceptable salt thereof comprises an HA hydrogel microsphere comprising crosslinked HA chains to which a plurality of drug moieties are covalently and reversibly conjugated, or a pharmaceutically acceptable salt thereof, wherein the drug conjugate comprises each of the following units:

[0460] [ka] [In the formula, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each R a1 , -R a2 , -L 2 - is used as defined elsewhere herein; Each -X'- has the formula (x0):

[0461] [ka] (In the formula, The unmarked dashed line indicates the bond to the carbonyl group, the dashed line with an asterisk indicates the bond to the sulfur atom, v0 is selected from the group consisting of 0 and 1; -X 1 -, -X 4 - is independently C 1~10 This C is an alkyl 1~10 Alkyl is -O-, -T-, -N(R y1 )-, -C(O)O- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1~10 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a Optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -R y2a -H and C 1~4 independently selected from the group consisting of alkyl, -X 2 - is -N(R 1 )—, —O—, —S—, and —Se—; =X 3 is =O, =N(R 1 ) and =S; -X 5 -C 1~20 This C is an alkyl 1~20 Alkyl is -O-, -C(O)O-, -T-, -N(R y1 )- and -N(R y1 )C(O)—, wherein the C 1~20 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a Optionally substituted with one or more groups independently selected from -R1 -H, C 1~5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 2 and optionally independently substituted with -R 2 is halogen, -CN, oxo, -C(O)OR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 )(R 3a ), -S(O)2N(R 3 )(R 3a ), -S(O)N(R 3 )(R 3a ), -S(O)R 3 , -S(O)R 3 , -N(R 3 )S(O)2N(R 3a )(R 3b ), -SR 3 , -N(R 3 )(R 3a ), -NO2, -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O)2R 3a , -N(R 3 )S(O)R 3a , -N(R 3 )C(O)OR 3a , -N(R 3 )C(O)N(R 3a )(R 3b ), -OC(O)N(R 3 )(R 3a ) and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 3 , -R 3a and -R 3b-H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. It is of Each -Y'- has the formula (y0):

[0462] [ka] (In the formula, The unmarked dashed line indicates the bond to the carbonyl group, and the dashed line with an asterisk indicates the bond to the nitrogen atom of the thiosuccinimide ring; -Y 1 -, -Y 4 - is independently C 1~10 This C is an alkyl 1~10 Alkyl is -O-, -T-, -N(R y1 )-, -C(O)O- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1~10 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a ) optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -R y2a is H and C 1~4 independently selected from the group consisting of alkyl, -Y 2 - is -N(R 2 )—, —O—, —S—, and —Se—; =Y 3 is =O, =N(R 2 ) and =S; -R 1 , -R 2 -H, C 1~5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from the group consisting of one or more -R 3 and optionally independently substituted with -R 3 is halogen, -CN, oxo, -C(O)OR 4 , -OR 4 , -C(O)R 4 , -C(O)N(R 4 )(R 4a ), -S(O)2N(R 4 )(R 4a ), -S(O)N(R 4 )(R 4a ), -S(O)R 4 , -S(O)R 4 , -N(R 4 )S(O)2N(R 4a )(R 4b ), -SR 4 , -N(R 4 )(R 4a ), -NO2, -OC(O)R 4 , -N(R 4 )C(O)R 4a , -N(R 4 )S(O)2R 4a , -N(R 4 )S(O)R 4a , -N(R 4 )C(O)OR 4a , -N(R 4 )C(O)N(R 4a )(R 4b ), -OC(O)N(R 4 )(R 4a ) and C 1~6 alkyl, C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 4 , -R 4a and -R 4b -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. It is of Each-L 1 - is a linker moiety of formula (I):

[0463] [ka] (In the formula, The dashed line indicates the attachment of -D to the nitrogen atom by forming an amide bond; -X- is -C(R 4 R 4a )-, -N(R 4 )-, -O-, -C(R 4 R 4a )-C(R 5 R 5a )-, -C(R 5 R 5a )-C(R 4 R 4a )-, -C(R 4 R 4a )-N(R 6 )-, -N(R 6 )-C(R 4 R 4a )-, -C(R 4 R 4a )-O-, -OC(R 4 R 4a )- or -C(R 7 R 7a )- and X 1 is C or S(O), -X 2 - is -C(R 8 R 8a )- or -C(R 8 R 8a )-C(R 9 R 9a )- and =X 3 is =O, =S, or =N-CN, -R 1 , -R 1a , -R 2 , -R 2a , -R 4 , -R 4a , -R 5 , -R 5a , -R 6 , -R8 , -R 8a , -R 9 , -R 9a -H, and C 1~6 independently selected from the group consisting of alkyl, -R 3 , -R 3a -H, and C 1~6 alkyl, with the proviso that -R 3 , -R 3a If one or both of are other than -H, they are sp 3 provided that it is connected to a bonded N through a hybridized carbon atom, -R 7 is -N(R 10 R 10a ), or -NR 10 -(C=O)-R 11 and -R 7a , -R 10 , -R 10a , -R 11 are, independently of each other, -H or C 1~10 is alkyl, Optionally, Pair-R 1a / -R 4a , -R 1a / -R 5a , -R 1a / -R 7a , -R 4a / -R 5a , -R 8a / -R 9a one or more of the following forms a chemical bond; Optionally, Pair-R 1 / -R 1a , -R 2 / -R 2a , -R 4 / -R 4a , -R 5 / -R 5a , -R 8 / -R 8a , -R 9 / -R 9a One or more of the following, together with the atoms to which they are attached, form a C 3~10 forming a cycloalkyl or a 3- to 10-membered heterocyclyl; Optionally, Pair-R 1 / -R 4 , -R 1 / -R 5 , -R 1 / -R 6 , -R 1 / -R 7a , -R 4 / -R 5 , -R 4 / -R 6 , -R 8 / -R 9 , -R 2 / -R 3 together with the atoms to which they are attached form ring A, Optionally, -R 3 / -R 3a together with the nitrogen atom to which they are attached form a 3- to 10-membered heterocyclic ring, Ring A is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl and Each-L 1 -Ha-L 2 -, with the proviso that the hydrogens marked with an asterisk in formula (I) are not replaced by a substituent. Contains multiple The drug conjugate has a Z in the range of about 50% to about 98% 1 , Z in the range of about 0.1% to about 20% 2 -i, Z in the range of about 0.1% to about 20% 3 -i and Z in the range of about 0.1% to about 10% 4 Includes -i.

[0464] Preferably, in formula (I), the -R 11 Ha-L 2 - is replaced by

[0465] In particular, the drug conjugates have a Z in the range of about 86% to about 96%. 1 , Z in the range of about 0.1% to about 12.9%2 -i, Z in the range of about 0.34% to about 3.57% 3 -i and Z in the range of about 0.1% to about 9.5% 4 Includes -i.

[0466] In certain embodiments, the drug conjugate or pharmaceutically acceptable salt thereof comprises an HA hydrogel microsphere comprising crosslinked HA chains to which a plurality of drug moieties are covalently and reversibly conjugated, or a pharmaceutically acceptable salt thereof, wherein the drug conjugate comprises each of the following units:

[0467] [ka] [In the formula, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each R a1 is H or an alkali metal ion, e.g., Na + and Each-R a2 is -H, Each -D is a ranibizumab moiety, Each -X'- has the formula (x4):

[0468] [ka] (wherein the unmarked dashed line indicates the bond to the carbonyl group, the dashed line with an asterisk indicates the bond to the sulfur atom, and c0 is 7.) It is of Each -Y'- has the formula (y4):

[0469] [ka] (wherein the unmarked dashed line indicates the bond to the carbonyl group and the dashed line marked with an asterisk indicates the bond to the nitrogen atom of the thiosuccinimide ring). It is of Each-L 2 -L 1 - is the formula (s1):

[0470] [ka] where the dashed line indicates the bond to the sulfur atom and the dashed line with an asterisk indicates the bond of -D to the nitrogen atom by forming an amide bond. [It is of the] Contains multiple The drug conjugates have a Z in the range of about 78% to about 96%. 1 , Z in the range of about 2% to about 10% 2 -i, Z in the range of about 1% to about 7% 3 -i and Z in the range of about 0.5% to about 5% 4 Includes -i.

[0471] Preferably, the drug conjugate, or a pharmaceutically acceptable salt thereof, comprises an HA hydrogel microsphere, or a pharmaceutically acceptable salt thereof, comprising crosslinked HA chains to which a plurality of drug moieties are covalently and reversibly conjugated, wherein the drug conjugate comprises each of the following units:

[0472] [ka] [In the formula, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each R a1 is H or an alkali metal ion, e.g., Na + and Each-R a2 is -H, Each -D is a ranibizumab moiety, Each -X'- has the formula (x4):

[0473] [ka] (wherein the unmarked dashed line indicates the bond to the carbonyl group, the dashed line with an asterisk indicates the bond to the sulfur atom, and c0 is 7.) It is of Each -Y'- has the formula (y4):

[0474] [ka] (wherein the unmarked dashed line indicates the bond to the carbonyl group and the dashed line marked with an asterisk indicates the bond to the nitrogen atom of the thiosuccinimide ring). It is of Each-L 2 -L 1 - is the formula (s2):

[0475] [ka] where the dashed line indicates the bond to the sulfur atom and the dashed line with an asterisk indicates the bond of -D to the nitrogen atom by forming an amide bond. [It is of the] Contains multiple The drug conjugate had a Z of approximately 92.9% 1 , about 4.3% Z 2 -i, about 1.5% Z 3 -i and approximately 1.3% Z 4 Includes -i.

[0476] Suitably, the drug conjugate depicted above has a Z of 91.82%. 1 , 5.29% Z 2 -i, 1.5% Z 3 -i and 1.39% Z 4 Includes -i.

[0477] Another aspect of the present invention is a pharmaceutical composition comprising a drug conjugate of the present invention, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0478] The present invention also relates to the drug conjugate of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for use as a pharmaceutical.

[0479] The present invention also relates to a drug conjugate of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in reducing or inhibiting angiogenesis in a subject having a disorder associated with pathological angiogenesis.

[0480] Another aspect of the present invention is a drug conjugate, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention for use in the treatment of a disorder associated with pathological angiogenesis.

[0481] In another aspect, the present invention relates to a drug conjugate or a pharmaceutically acceptable salt thereof for use in treating a disorder associated with pathological angiogenesis, such as an ocular disorder, wherein a single intraocular administration of the drug conjugate or the pharmaceutically acceptable salt thereof provides an intravitreal therapeutically effective amount of a VEGF-neutralizing drug for at least six months.

[0482] The present invention also relates to a drug conjugate or a pharmaceutically acceptable salt thereof for use in treating a disorder associated with pathological angiogenesis, such as an ocular disorder, wherein a single intraocular administration of the drug conjugate or the pharmaceutically acceptable salt thereof provides an intravitreal therapeutically effective amount of ranibizumab of at least 0.7 μg / ml for at least 6 months.

[0483] Disorders associated with pathological angiogenesis include age-related macular degeneration (AMD), macular degeneration, macular edema, diabetic macular edema (DME), retinopathy, diabetic retinopathy (DR), other ischemia-related retinopathies, retinopathy of prematurity (ROP), retinal vein occlusion (RVO), CNV, corneal neovascularization, diseases associated with corneal neovascularization, retinal neovascularization, diseases associated with retinal / choroidal neovascularization, pathological myopia, von Hippel-Lindau disease, ocular histoplasmosis, familial exudative vitreoretinopathy (FEVR), Coats' disease, Norrie's disease, and osteoporosis. The eye disorder may be an eye disorder selected from the group consisting of pseudoglioma syndrome (OPPG), subconjunctival hemorrhage, rubeosis, ocular neovascular disease, neovascular glaucoma, retinitis pigmentosa (RP), hypertensive retinopathy, retinal angiomatous proliferation, macular telangiectasia, iris neovascularization, intraocular neovascularization, retinal degeneration, cystoid macular edema (CME), vasculitis, papilledema, retinitis, conjunctivitis, Leber's congenital amaurosis, uveitis, choroiditis, ocular histoplasmosis, blepharitis, dry eye, traumatic eye injury, and Sjogren's disease.

[0484] Exemplary diseases associated with corneal neovascularization may be selected from the group consisting of epidemic keratoconjunctivitis, vitamin A deficiency, contact lens overwear, atopic keratitis, superior limbal keratitis, pterygium, keratitis sicca, Sjogren's syndrome, acne rosacea, phlyctenosis, syphilis, mycobacterial infection, lipid degeneration, chemical burns, bacterial ulcers, fungal ulcers, herpes simplex infection, herpes zoster infection, protozoal infection, Kaposi's sarcoma, Mooren's ulcer, Therrien's peripheral corneal dystrophy, marginal keratolysis, rheumatoid arthritis, systemic lupus, polyarteritis nodosa, trauma, Wegener's sarcoidosis, scleritis, Stevens-Johnson syndrome, pemphigoid, radial keratotomy, and corneal graft rejection.

[0485] Exemplary diseases associated with retinal / choroidal neovascularization may be selected from the group consisting of diabetic retinopathy, macular degeneration, sickle cell anemia, sarcoid, syphilis, pseudoxanthoma elasticum, Paget's disease, venous occlusion, arterial occlusion, carotid occlusive disease, chronic uveitis / vitreous inflammation, mycobacterial infections, Lyme disease, systemic lupus erythematosus, retinopathy of prematurity, retinitis pigmentosa, retinal edema, Eales' disease, Behcet's disease, infections causing retinitis or choroiditis, presumed ocular histoplasmosis, Best's disease, myopia, optic pit, Stargardt's disease, pars planitis, retinal detachment, hyperviscosity syndrome, toxoplasmosis, trauma, and post-laser complications.

[0486] In certain embodiments, the ocular disorder is selected from the group consisting of AMD, DME, DR, and RVO.

[0487] Suitably, the eye disorder is AMD, such as wet AMD.

[0488] The drug conjugate or pharmaceutically acceptable salt thereof or pharmaceutical composition of the present invention may be administered intravitreally, for example, by intravitreal injection, by eye drops, intramuscularly, topically, subconjunctivally, intravesicularly, intraocularly, intraorbitally, by injection, by implantation or by infusion.

[0489] The drug conjugate of the present invention or a pharmaceutically acceptable salt thereof may be administered by intraocular administration, for example, by intraocular administration into the vitreous of a subject, every 6 months, every 9 months, or every 12 months.

[0490] Preferably, the drug conjugate of the present invention, or a pharmaceutically acceptable salt thereof, is administered every six months by intraocular administration into the vitreous of the subject.

[0491] The present invention also relates to the drug conjugate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention for use in the manufacture of a medicament for the treatment of wet AMD.

[0492] Another aspect of the present invention relates to a kit of parts comprising a drug conjugate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention.

[0493] The present invention also relates to a pre-filled syringe comprising the drug conjugate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention.

[0494] Another aspect of the present invention relates to a pharmaceutical composition comprising a combination of a drug conjugate of the present invention, or a pharmaceutically acceptable salt thereof, and at least one additional drug, for use in treating an ocular disorder.

[0495] The present invention also relates to a pharmaceutical composition comprising a drug conjugate of the present invention, or a pharmaceutically acceptable salt thereof, for use in combination therapy of an ocular disorder with at least one additional drug.

[0496] In certain embodiments, at least one additional drug is a drug in free form. In certain embodiments, at least one additional drug is in the form of a stable conjugate. In certain embodiments, at least one additional drug is in the form of a controlled release compound.

[0497] In certain embodiments, the at least one additional drug is selected from the group consisting of proteins, polypeptides, antibodies, anti-angiogenic agents, cytokines, cytokine antagonists, corticosteroids, and analgesics.

[0498] In certain embodiments, the at least one additional drug is selected from the group consisting of a complement pathway inhibitor, a Tie2 pathway stimulator, a BCL-xL inhibitor, an integrin receptor / integrin antagonist, a Rho kinase inhibitor, a human protein tyrosine phosphatase beta inhibitor, a fibroblast growth factor inhibitor, a chemokine receptor type 3 antagonist, a connexin 43 inhibitor, a plasma kallikrein inhibitor, a Ref-1 inhibitor, a matrix metalloproteinase inhibitor, an MBL-related serine protease inhibitor, an NLRP3 inflammasome, an HtrA1 inhibitor, a mitochondrial targeting agent, a vitamin A substitute, a steroid, an immunosuppressant, a prostaglandin compound, a beta blocker, an alpha adrenergic agonist, a carbonic anhydrase inhibitor, a miotic or cholinergic agent, and epinephrine.

[0499] Exemplary complement pathway inhibitors may be selected from the group consisting of C3 inhibitors, such as APL-1, APL-2, compstatin and its analogs, IBI-302 or KNP-301; C5 inhibitors, such as Zimura or PAS-Nomacopan; CFB inhibitors, such as RG-6299; C1q inhibitors, such as ANX007; CFH, such as GEM-103 or AVTS-001; CFD, such as Danicopan (ALXN2040); bispecific C3b and CD59 inhibitors, such as KNP-302; CD59 inhibitors, such as JNJ-1887 and CFI, such as GT-005.

[0500] Exemplary Tie2 pathway stimulators include Ang1 mimetics such as AV-001 (Vasculotide), EG-Mirotin, AP-102, AXT-107 (Gersizangitide), Man-01, Man-11, PMC-402, or UBB-2000 (UBX-2050) and Ang2 inhibitors such as Nesvacumab, Zansecimab (LY-3127804), PF-4856884, MEDI- 3617, Faricimab, BI836880, CVX-241, Dutafab anti-VEGF / ang2, Vanucizumab, ASKG-712, RG-6120, trebananib, Xanteximab, ABP-201 / 200, DR-30121, PMC-401, PMC-404, RBD-5078, RO-101, RO-634LQ-016, LQ-017, AMG-780, AT-066, Atu-111 or CVX-060.

[0501] In certain embodiments, the BCL-xL inhibitor is UBX1325.

[0502] Exemplary integrin receptor / integrin antagonists may be selected from the group consisting of luminate, SF0166, AXT-107 (gelcizangitide) and THR687.

[0503] Exemplary Rho kinase inhibitors may be selected from the group consisting of Ripasudil (K-115) and Netarsudil (AR-13503).

[0504] In certain embodiments, the human protein tyrosine phosphatase beta inhibitor is Razuprotafib (AKB-9778).

[0505] In certain embodiments, the fibroblast growth factor inhibitor is X-82.

[0506] Exemplary chemokine receptor type 3 antagonists may be selected from the group consisting of GW766994X and GW782415X.

[0507] In certain embodiments, the connexin 43 inhibitor is HCB1019.

[0508] Exemplary plasma kallikrein inhibitors may be selected from the group consisting of KVD001 and THR-149.

[0509] In certain embodiments, the Ref-1 inhibitor is APX3330.

[0510] In certain embodiments, the matrix metalloproteinase inhibitor is AG3340.

[0511] Exemplary MBL-associated serine protease inhibitors may be selected from the group consisting of narsoplimab and OMS906.

[0512] In certain embodiments, the NLRP3 inflammasome is Xiflam.

[0513] In certain embodiments, the HtrA1 inhibitor has an IC-500.

[0514] Exemplary mitochondrial targeting agents may be selected from the group consisting of Elamipretide, Visomitin, and MC-16.

[0515] In one particular embodiment, the vitamin A substitute is ALK-001.

[0516] Exemplary steroids may be selected from the group consisting of dexamethasone, fluocinolone acetonide, and prednisolone.

[0517] Exemplary immunosuppressants include antimetabolites, such as azathioprine, methotrexate, or mycophenolate; calcineurin inhibitors, such as cyclosporine or tacrolimus; alkylating agents, such as cyclophosphamide or chlorambucil; monoclonal antibodies; antibody fusion proteins or fragments, such as Fab, F(ab')2, or ScFv, including adalimumab, infliximab, etanercept, certolizumab, canakinumab, gevokizumab, daclizumab, tocilizumab, sarilimumab, ab), KSI-501, EBI-301, secukinumab, rituximab, alemtuzumab, interferon alpha 2a or 2b or interferon beta, and JAK inhibitors such as ruxolitinib, tofacitinib, oclacitinib, filgotinib, baricitinib, peficitinib, upadacitinib, fedratinib, abrocitinib, pacritinib, deuclavacitinib, cerdulatinib, gandotinib, lestaurtinib or momelotinib.

[0518] Exemplary prostaglandin compounds may be selected from the group consisting of latanoprost, bimatoprost, and travoprost.

[0519] Exemplary beta blockers may be selected from the group consisting of timolol, betaxolol, and metipranolol.

[0520] Exemplary alpha adrenergic agonists may be selected from the group consisting of apraclonidine and brimonidine.

[0521] Exemplary carbonic anhydrase inhibitors may be selected from the group consisting of dorzolamide and brinzolamide.

[0522] Exemplary miotics or cholinergic agents may be selected from the group consisting of pilocarpine and carbachol.

[0523] In certain embodiments, the epinephrine is dipivefrin.

[0524] In certain embodiments, the drug conjugate of the present invention, or a pharmaceutically acceptable salt thereof, and at least one additional drug are formulated for simultaneous administration.

[0525] In certain embodiments, the drug conjugate of the present invention, or a pharmaceutically acceptable salt thereof, and at least one additional drug are formulated for separate administration.

[0526] The present invention also refers to a method for treating an eye disorder, comprising the step of administering a therapeutic amount of the drug conjugate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention to a subject in need of treatment for an eye disorder.

[0527] The present invention also relates to a method of treating an ocular disorder, comprising administering to a subject in need of treatment for the ocular disorder a therapeutic amount of a pharmaceutical composition comprising a combination of a drug conjugate or a pharmaceutically acceptable salt thereof and at least one additional drug.

[0528] The present invention also relates to a method comprising the step of administering to a subject in need thereof a therapeutic amount of a pharmaceutical composition comprising the drug conjugate of the present invention or a pharmaceutically acceptable salt thereof, wherein said composition can be used in combination therapy with at least one additional drug.

[0529] Administration can be by intraocular administration, for example, by intraocular injection into the vitreous of the subject.

[0530] Exemplary ocular disorders include age-related macular degeneration (AMD), macular degeneration, macular edema, diabetic macular edema (DME), retinopathy, diabetic retinopathy (DR), other ischemia-related retinopathies, retinopathy of prematurity (ROP), retinal vein occlusion (RVO), CNV, corneal neovascularization, diseases associated with corneal neovascularization, retinal neovascularization, uveitis macular edema, branch retinal vein occlusion (BRVO), central retinal vein occlusion (CRVO), submacular hemorrhage, polypoidal choroidal vasculopathy (PCV), retinal capillary microaneurysm, retinal artery occlusion (RAO), branch retinal artery occlusion (BRAO), central retinal artery occlusion (CRAO), subfoveal hemorrhage, subretinal hemorrhage, radiation retinopathy, exudative retinal detachment, Eales' disease, neovascular macular telangiectasia, ischemic retinal vasculitis, diseases associated with retinal or choroidal neovascularization, pathologic myopia, von Hippel-Lindau disease, ocular histoplasmosis, familial exudative vitreoretinopathy (FEVR), Coats' disease, Norrie's disease, osteoporosis-pseudoglioma syndrome (OPPG), subconjunctival hemorrhage, rubeosis, ocular neovascular diseases, neovascular glaucoma, retinitis pigmentosa (RP), hypertensive retinopathy, retinal angiomatous proliferation, macular telangiectasia, iris neovascularization, intraocular neovascularization, retinal degeneration, cystoid macular edema (CME), vasculitis, papilledema, retinitis, conjunctivitis, Leber's congenital amaurosis, uveitis, choroiditis, ocular histoplasmosis, blepharitis, dry eye, traumatic eye injury, and Sjogren's disease.

[0531] Eye disorders also include age-related macular degeneration (AMD), macular degeneration, macular edema, diabetic macular edema (DME), retinopathy, diabetic retinopathy (DR), other ischemia-related retinopathies, retinopathy of prematurity (ROP), retinal vein occlusion (RVO), CNV, corneal neovascularization, diseases associated with corneal neovascularization, retinal neovascularization, diseases associated with retinal or choroidal neovascularization, pathological myopia, von Hippel-Lindau disease, ocular histoplasmosis, familial exudative vitreoretinopathy (FEVR), Coats disease, Norrie disease, bone The ocular neovascular disease may be selected from the group consisting of opioid pseudoglioma syndrome (OPPG), subconjunctival hemorrhage, rubeosis, ocular neovascular disease, neovascular glaucoma, retinitis pigmentosa (RP), hypertensive retinopathy, retinal angiomatous proliferation, macular telangiectasia, iris neovascularization, intraocular neovascularization, retinal degeneration, cystoid macular edema (CME), vasculitis, papilledema, retinitis, conjunctivitis, Leber's congenital amaurosis, uveitis, choroiditis, ocular histoplasmosis, blepharitis, dry eye, traumatic eye injury, and Sjogren's disease.

[0532] In certain embodiments, the ocular disorder is selected from the group consisting of AMD, DME, DR, and RVO.

[0533] Suitably, the eye disorder is AMD, such as wet AMD.

[0534] Another aspect of the present invention relates to a method for preparing a drug conjugate or a pharmaceutically acceptable salt thereof, comprising hydrogel HA microspheres or a pharmaceutically acceptable salt thereof comprising cross-linked HA chains. The hydrogel HA microspheres or a pharmaceutically acceptable salt thereof can be advantageously used as a drug carrier because they are physiologically tolerable.

[0535] Accordingly, the present invention also provides a method for preparing a drug conjugate or a pharmaceutically acceptable salt thereof, comprising: (a) mixing solution A with solution B to form an emulsion, wherein solution A comprises a first functionalized HA modified with one or more -FG1 and optionally further functional groups, and a second functionalized HA modified with one or more -FG2 and optionally further functional groups, -FG1 and -FG2 being different functional moieties, and -FG1 on the first functionalized HA reacts with -FG2 on the second functionalized HA to form multiple crosslinks resulting in the formation of hydrogel HA microspheres; (b) optionally adding a pH adjuster to the emulsion of step (a); (c) collecting the resulting hydrogel HA microspheres of step (a) or (b); (d) providing the hydrogel HA microspheres or pharmaceutically acceptable salts thereof of step (c), wherein the hydrogel contains one or more unreacted -FG1 or -FG2; (e) Monoconjugate Reagent DL 1 -L 2 -FG3, bisconjugate reagent FG3-L 2 -L 1 -DL 1 -L 2 -FG3, or a Tris conjugate reagent of formula (t):

[0536] [ka] [Wherein -D independently represents -L 1 a VEGF-neutralizing drug moiety covalently and reversibly conjugated to Each-L 1 - is independently a reversible linker moiety; Each-L 2 - is independently a spacer moiety or absent; Each -FG3 is independently a functional group that reacts with -FG1 or with -FG2. preparing a (f) mixing the hydrogel HA microspheres of step (d) with the monoconjugate, bisconjugate, or trisconjugate reagent of step (e); (g) optionally, mixing the drug conjugate or pharmaceutically acceptable salt thereof of step (f) with a blocking reagent; and (h) collecting the drug conjugate or pharmaceutically acceptable salt thereof of step (f) or (g). The present invention relates to a method, comprising:

[0537] The present invention also relates to the above method, wherein step (b) is not optional and the first and second functionalized HA are not modified with additional functional groups.

[0538] Throughout this specification, -L 3 It is understood that - is the linkage resulting from the reaction of -FG1 with -FG2.

[0539] The polymerization in the method of the present invention occurs by suspension polymerization. In step (a), solutions A and B form an emulsion, and after sufficient mixing time, solution A becomes the dispersed phase, while solution B becomes the continuous phase. The dispersed phase should be immiscible with the continuous phase. Also, in step (a), both functionalized HAs are mainly or exclusively linear HA chains.

[0540] In step (a), the mixing of solution A and solution B to form the emulsion may require vigorous stirring, pressure or other forces, which may be achieved by stirring, for example by stirring using a pitched blade stirrer in combination with a baffle; by shaking, for example by shaking in a container such as a Falcon tube (sealed tube); by using a rotor or stator; by using an ultrasonic device; by using a static mixer, for example by using a packed bead column or a flow plate; by using a membrane with defined pores, for example by using cross-flow membrane emulsification techniques (in which a material in liquid form to be formed into microparticles is forced through a membrane containing microscale pores to form a flowing solution of the dispersed phase) or stirred-cell membrane emulsification; by using a tubular membrane with a surface made of hydrophobic plastic, such as that disclosed in WO 2022 / 198052 A2, the entirety of which is incorporated herein by reference; by using a microfluidic droplet generator; or by spray polymerization in air, for example by using an ultrasonic atomizer.

[0541] Suitably, the mixing in step (a) is achieved by stirring, for example by stirring using a pitched blade stirrer in combination with a baffle, or by using a microfluidic droplet generator.

[0542] In certain embodiments, all -FG1's are the same and all -FG2's are the same.

[0543] Suitably, the first and second functionalised HA of the method of the invention are not optionally modified with further functional groups.

[0544] The method of the present invention optionally further comprises the step of size fractionating the resulting hydrogel microspheres of step (c) to obtain microspheres having a particular particle size distribution.

[0545] It will also be recognized by those skilled in the art that, throughout this specification, there may be optional washing or purification steps between any steps of the methods of the present invention. Specifically, the methods may further include optional washing or purification steps of the hydrogel or drug conjugate obtained in step (c), (f), or (g).

[0546] In certain embodiments, the method of the present invention comprises: (a) mixing solution A with solution B to form an emulsion, wherein solution A comprises a first functionalized HA modified with one or more -FG1 and optionally further functional groups, and a second functionalized HA modified with one or more -FG2 and optionally further functional groups, -FG1 and -FG2 being different functional moieties, and -FG1 on the first functionalized HA reacts with -FG2 on the second functionalized HA to form multiple crosslinks resulting in the formation of hydrogel HA microspheres; (b) optionally adding a pH adjuster to the emulsion of step (a); (c) collecting the resulting hydrogel HA microspheres of step (a) or (b); (d) optionally, size fractionating the resulting hydrogel HA microspheres of step (a), (b), or (c) to obtain microspheres having a particular particle size distribution; (e) optionally washing the microspheres obtained in step (a), (b), (c) or (d); (f) optionally, collecting the hydrogel HA microspheres of step (d) or (e); (g) providing the hydrogel HA microspheres or pharmaceutically acceptable salts thereof of step (c) or (f), wherein the hydrogel contains one or more unreacted -FG1 or -FG2; (h) Monoconjugate Reagent DL 1 -L 2 -FG3, bisconjugate reagent FG3-L 2 -L 1 -DL1 -L 2 -FG3, or a Tris conjugate reagent of formula (t):

[0547] [ka] [In the formula, -D, each -L 1 - and -L 2 - is used as described elsewhere herein; Each -FG3 is independently a functional group that reacts with -FG1 or with -FG2. preparing a (i) mixing the hydrogel HA microspheres of step (g) with the monoconjugate, bisconjugate or trisconjugate reagent of step (h); (j) optionally, mixing the drug conjugate of step (i) or a pharmaceutically acceptable salt thereof with a blocking reagent; and (k) collecting the drug conjugate or pharmaceutically acceptable salt thereof of step (i) or (j). Includes.

[0548] In certain embodiments, the molecular weights of the first and second functionalized HA are independently in the range of about 80 kDa to about 250 kDa, e.g., about 90 kDa to about 200 kDa, or about 100 kDa to about 150 kDa. Preferably, the molecular weights of the first and second functionalized HA are in the range of 100 kDa to 150 kDa. It is understood that the HA may be polydisperse and contain polymer chains of unequal lengths; therefore, molecular weight is not a single value; i.e., the HA exists as a distribution of chain lengths and molecular weights. For example, if the molecular weight of the first or second functionalized HA is 125 kDa, the HA may contain polymer HA chains in the range of about 30 kDa to about 400 kDa.

[0549] In certain embodiments, in step (d), the microspheres obtained have a diameter in the range of about 1 μm to about 1000 μm, such as about 50 μm to about 900 μm, such as about 100 μm to about 700 μm, such as about 200 μm to about 500 μm, or such as about 50 μm to about 500 μm, as determined by flow microscopy or other similar methods known in the art. In certain embodiments, in step (d), the microspheres obtained have a diameter in the range of 1 μm to 1000 μm, such as 50 μm to 900 μm, such as 100 μm to 700 μm, such as 200 μm to 500 μm, or such as 50 μm to 500 μm, as determined by flow microscopy. In certain embodiments, in step (d), the microspheres obtained have a diameter in the range of 50 μm to 500 μm, as determined by flow microscopy. In certain embodiments, in step (d), the resulting microspheres have diameters in the range of 100 μm to 200 μm as determined by flow microscopy.

[0550] Preferably, in step (d), the obtained microspheres have a diameter of ≧50 μm as determined by flow microscopy. 10 Value and d≦900 μm 90 value, e.g., d ≥ 100 μm 10 Value and d≦700 μm 90 Value or d ≥ 200 μm 10 Value and d≦500 μm 90 In certain embodiments, in step (d), the resulting microspheres have a d value of ≧100 μm. 10 Value and d≦200 μm 90 Preferably, for these measurements, the microspheres are stored in succinate buffer.

[0551] As used herein, the parameter d 10 The value represents the point in the size distribution below which 10% of the total amount of material in the sample is contained. 90The value is the size below which 90% of the mass of the substance is contained. The swelling of HA microspheres can be affected by the buffer in which the microspheres are stored during the measurement, and / or by the pH, osmolality, and ionic strength, and therefore this is d 10 value and d 90 It is understood that this may affect the values.

[0552] In certain embodiments, the method of the present invention comprises: (a) mixing solution A with solution B to form an emulsion, wherein solution A comprises a first functionalized HA modified with one or more -FG1 and optionally further functional groups, and a second functionalized HA modified with one or more -FG2 and optionally further functional groups, -FG1 and -FG2 being different functional moieties, and -FG1 on the first functionalized HA reacts with -FG2 on the second functionalized HA to form multiple crosslinks resulting in the formation of hydrogel HA microspheres; (b) adding a pH adjuster to the emulsion of step (a); (c) collecting the resulting hydrogel HA microspheres of step (b); (d) size fractionating the resulting hydrogel HA microspheres of step (c) to obtain microspheres having a specific particle size distribution; (e) optionally washing the microspheres obtained in step (c) or (d); (f) harvesting the hydrogel HA microspheres of step (d) or (e); (g) providing the hydrogel HA microspheres or pharmaceutically acceptable salts thereof of step (f), wherein the hydrogel contains one or more unreacted -FG1 or -FG2; (h) Monoconjugate Reagent DL 1 -L 2 -FG3, bisconjugate reagent FG3-L 2 -L 1 -DL 1 -L2 -FG3, or a Tris conjugate reagent of formula (t):

[0553] [ka] [In the formula, -D, each -L 1 - and -L 2 - is used as described elsewhere herein; Each -FG3 is independently a functional group that reacts with -FG1 or with -FG2. preparing a (i) mixing the hydrogel HA microspheres of step (g) with the monoconjugate, bisconjugate or trisconjugate reagent of step (h); (j) optionally, mixing the drug conjugate of step (i) or a pharmaceutically acceptable salt thereof with a blocking reagent; and (k) collecting the drug conjugate or pharmaceutically acceptable salt thereof of step (i) or (j). Includes.

[0554] The present invention also relates to a drug conjugate or a pharmaceutically acceptable salt thereof obtainable by the method of the present invention.

[0555] Those skilled in the art will recognize that the drug conjugates of the present invention, or pharmaceutically acceptable salts thereof, may also contain one or more unreacted -FG1 or -FG2 groups. A blocking reagent is used to avoid undesired reactions between these groups and other functional groups on the drug moiety or compound that may be found where the drug conjugate is administered.

[0556] Exemplary blocking reagents include:

[0557] [ka] may be selected from the group consisting of:

[0558] More specifically, to avoid undesired reactions between unreacted maleimides on HA hydrogel microspheres and nucleophilic compounds such as thiols, particularly glutathione, or functional groups on drug moieties such as amines, the unreacted maleimides are reacted with a blocking reagent. Advantageously, the blocking reagent of formula (r01) inhibits the retro-Michael and exchange reactions of the formed thiosuccinimides in the presence of other thiol-containing compounds at physiological pH and temperature. This is particularly beneficial for drug conjugates or pharmaceutically acceptable salts thereof administered to tissues or organs where glutathione is naturally found, such as the eye.

[0559] In certain embodiments, the reagent in step (e) is a monoconjugate reagent DL 1 -L 2 In certain embodiments, the reagent in step (e) is the bisconjugate reagent FG3-L 2 -L 1 -DL 1 -L 2 -FG3. In certain embodiments, the reagent in step (e) is a tris-conjugate reagent of formula (t).

[0560] Monoconjugate Reagent DL 1 -L 2 -FG3 can be obtained by any method known in the art. This is followed by purification by tagging the resulting drug-reversible prodrug linker conjugate with a purification tag to form a mixture. Monoconjugate Reagent DL 1 -L 2 -FG3 is purified from the mixture by chromatographic separation, and the purification tag is then added to the tagged DL 1 -L 2 -Removed from FG3 and purified DL 1 -L 2 - Form FG3.

[0561] As used herein, a "purification tag" refers to a moiety that, when conjugated to a second moiety, confers physical and / or chemical property(ies) that are not present in said second moiety without the tag moiety, and which different physical and / or chemical property(ies) allow for the purification of such a conjugate.

[0562] Purification tags within the scope of the present disclosure are described in WO 2015 / 052155 A1, which is incorporated herein by reference in its entirety. Thus, in certain embodiments, the purification tag comprises a moiety of formula (ax):

[0563] [ka] [wherein the dashed line represents DL via -FG3] 1 -L 2 -FG3, -R 1 , -R 1a , -R 1b , -R 2 , -R 2a , -R 2b , -R 3 , -R 3a , -R 3b , -R 4 , -R 4a , -R 4b are, independently of each other, H or methyl, each m is, independently of the other, 1, 2, 3, 4, 5, 6, 7, or 8; each n is, independently of the other, 1, 2, 3, 4, 5, 6, 7, or 8; each x is, independently of the other, 1, 2, 3, 4, 5, 6, 7, or 8; each y is, independently of the other, 0, 1, 2, 3, 4, 5, 6, 7, or 8; Includes.

[0564] In certain embodiments, the reagent in step (e) is a monoconjugate reagent DL of formula (m1): 1 -L 2 -FG3:

[0565] [ka] wherein -D is used as described elsewhere herein. is.

[0566] As used herein, and throughout the specification, for convenience, most structures do not show the stereochemistry and therefore represent all possible stereoisomers.

[0567] In certain embodiments, the reagent in step (e) is a monoconjugate reagent DL of formula (m'1): 1 -L 2 -FG3:

[0568] [ka] wherein -D is used as described elsewhere herein. is.

[0569] Surprisingly, it has been found that the drug conjugates of the present invention, or pharmaceutically acceptable salts thereof, use HA microspheres as carriers, which are effectively crosslinked, i.e., synthesized from linear, functionalized HA chains with a low degree of substitution. Furthermore, it has been observed that the size of the microspheres significantly improves the injectability of drug conjugates containing hydrogel HA microspheres compared to the injectability of coherent gels, such as the coherent gel disclosed in WO 2018 / 175788 A1. Another advantage of using HA hydrogel microspheres as carriers for drug conjugates is that the microspheres can be washed before administration, thus easily removing soluble by-products. On the other hand, the coherent gel disclosed in WO 2018 / 175788 A1 is obtained by polymerization in a syringe and then injected directly into the eye, and therefore cannot undergo a washing step. Furthermore, the coherent gels disclosed in WO 2018 / 175788 A1 tend to harden while being filled into syringes, which makes the syringe filling process more difficult. In contrast, the HA hydrogel microspheres used as carriers in the present invention can be stored after synthesis and then easily filled into final containers or further conjugated to drug moieties at a later time.

[0570] Preferably, solution A in step (a) comprises first and second functionalized HA and a solvent in which said functionalized HA can be dissolved.

[0571] In certain embodiments, solution A in step (a) comprises first and second functionalized HA and dimethyl sulfoxide, DMF, DMA, or a mixture thereof. In certain embodiments, solution A in step (a) comprises first and second functionalized HA and dimethyl sulfoxide.

[0572] In certain embodiments, solution A in step (a) comprises first and second functionalized HA, dimethyl sulfoxide, and water. In certain embodiments, solution A in step (a) comprises first and second functionalized HA, dimethyl sulfoxide, and a buffer.

[0573] In certain embodiments, solution A in step (a) is an aqueous solution and comprises the first and second functionalized HA and a buffering agent.

[0574] In general, it will be apparent to one skilled in the art that the term "buffer" can refer to one buffer or a mixture of two or more buffers.

[0575] Exemplary buffering agents include N-(2-acetamido)-2-aminoethanesulfonic acid (ACES), acetate, 2,2′,2″-nitrilotriacetic acid (ADA), adipate, alanine, ammonium, 2-amino-2-methyl-1-propanol (AMP), 2-amino-2-methyl-1,3-propanediol (AMPD), N-(1,1-dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid (AMPSO), arginine, ascorbate, aspartate, benzoate, N,N-bis(2-hydroxyethyl)-2-azapropanesulfonic acid, and the like. Aminoethanesulfonic acid (BES), bicarbonate, N,N-bis(2-hydroxyethyl)glycine, bis-(2-hydroxyethyl)amino-tris(hydroxymethyl)-methane, 1,3-bis(tris(hydroxymethyl)methylamino)propane, borate, 4-(cyclohexylamino)butane-1-sulfonic acid (CABS), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), 3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid (CAPSO), carbonate, N-cyclohexyl-2-aminoethanesulfonic acid Acid (CHES), Citrate, Diethanolamine, 3-(N,N-Bis[2-hydroxyethyl]amino)-2-hydroxypropanesulfonic acid (DIPSO), Edetate, Ethanolamine, Ethylenediamine, Formate, Fumarate, Gluconate, Glutamate, Glycine, Glycylglycine, Guanidine, N-(2-Hydroxyethyl)piperazine-N'-(4-butanesulfonic acid) (HEPBS), 4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 3-[4-(2-hydroxyethyl)piperazine-1- [yl]propane-1-sulfonic acid (HEPPS), N-(2-hydroxyethyl)piperazine-N'-(propanesulfonic acid) (HEPPSO), histidine, hydrazine, imidazole, lactate, lysine, malate, maleate, 2-(N-morpholino)ethanesulfonic acid (MES), metaphosphate, methylamine, 4-(4-morpholinyl)butanesulfonic acid (MOBS), 3-(N-morpholino)propanesulfonic acid (MOPS), 2-hydroxy-3-morpholinopropanesulfonic acid (MOPSO), pentetate, phosphate, piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES), piperazine, piperidine, piperazine-N,N'-bis(2-hydroxypropanesulfonic acid) (POPSO), propionate, pyridine, pyrophosphate, pyruvate, sorbate, succinate, N-tris(hydroxymethyl)methyl-4-aminobutanesulfonic acid (TABS), ([tris(hydroxymethyl)methylamino]propanesulfonic acid (TAPS), 2-hydroxy-3-[tris(hydroxymethyl)methylamino]-1-propanesulfonic acid (TAPSO), tartrate, taurine, 2-{[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino}ethane-1-sulfonic acid (TES), tricine, triethanolamine, tromethamine, and α-ketoglutarate.

[0576] It will be apparent to one skilled in the art that the corresponding conjugate acids, bases or salts of the buffering agents, as well as mixtures thereof, are also included.

[0577] In certain embodiments, the aqueous solution comprises first and second functionalized HA and a buffer, such as a buffer selected from the group consisting of citrate and histidine or a mixture thereof. In certain embodiments, the buffer comprises a mixture of citrate and histidine. In certain embodiments, the buffer consists of a mixture of citrate and histidine.

[0578] As defined herein, the term "histidine" is intended to encompass both D-histidine and L-histidine, as well as mixtures thereof. In certain embodiments, the term "histidine" refers to L-histidine. In certain embodiments, the term "histidine" refers to D-histidine. In certain embodiments, the term "histidine" refers to a mixture of L-histidine and D-histidine.

[0579] In certain embodiments, the buffering agent is L-histidine.

[0580] In certain embodiments, the buffer consists of a mixture of citrate, L-histidine, and sodium chloride.

[0581] In one particular embodiment, the aqueous solution comprises first and second functionalized HA, citrate, and sodium chloride.

[0582] In certain embodiments, the aqueous solution consists of first and second functionalized HA, citrate, and sodium chloride. In certain embodiments, the aqueous solution comprises first and second functionalized HA, histidine, and sodium chloride.

[0583] In certain embodiments, the aqueous solution comprises the first and second functionalized HA and an emulsifier, while solution B comprises a solvent.

[0584] The buffering agent may generally be added in an amount of about 0.01 mM to about 500 mM. In certain embodiments, the buffering agent has a concentration ranging from about 0.5 mM to about 350 mM. In certain embodiments, the buffering agent has a concentration ranging from about 1 mM to about 250 mM. In certain embodiments, the buffering agent has a concentration ranging from about 5 mM to 100 mM. In certain embodiments, the buffering agent has a concentration of about 100 mM. In certain embodiments, the buffering agent has a concentration of 100 mM. In certain embodiments, the buffering agent has a concentration of about 5 mM. In certain embodiments, the buffering agent has a concentration of 5 mM.

[0585] Preferably, solution B in step (a) comprises an emulsifier and a solvent.

[0586] In general, it will be apparent to those skilled in the art that the term "solvent" can refer to one solvent or a mixture of two or more solvents, and that the term "emulsifier" can refer to one emulsifier or a mixture of two or more emulsifiers.

[0587] Exemplary emulsifiers are sorbitan esters, such as sorbitan monolaurate (Span® 20), sorbitan monooleate (Span® 80), sorbitan monopalmitate (Span® 40), sorbitan monostearate (Span® 60), sorbitan sesquioleate (Span® 83), sorbitan trioleate (Span® 85) or sorbitan tristearate (Span® 65); PEG-30 dipolyhydroxystearate (Cithrol™ DPHS); polyglyceryl-3 diisostearate; mixtures of sorbitan oleate with copolymer esters of hydroxystearic acid and ethylene glycol (Hypermer™ 1083); polyoxyethylene sorbitan monooleate (Polysorbate 80, Tween® 80 and Tween® 80R); alcohols such as propanol, butanol, pentanol, hexanol, heptanol or octanol; alkyl and arylamine salts, such as primary, quaternary, secondary or tertiary amine salts; alkyl dimethyl betaine; alkyl ethoxylate sulfate; alkyl phenyl polyoxyethylene ethers, such as Octoxynol 9, Triton X-100, Igepal™ or Nonidet® P40; alkyl phosphates, such as mono- or di-alkyl phosphates; alkyl polyoxyethylene ethers, such as oct ... Ethylene ethers, such as laureth-4, laureth-9, laureth-23, ceteth-2, ceteth-10, ceteth-20, ceteareth-6, ceteareth-20, ceteareth-25, steareth-2, steareth-10, steareth-20, oleth-2, oleth-10, oleth-20, deceth-10 or trideceth-10; alkyl sulfates, such as sodium dodecyl sulfate (SDS); alkyl xanthates; bile salts, such as sodium cholate or sodium deoxycholate;Cationic lipids, such as cetyltrimethylammonium bromide, cetyltrimethylammonium chloride, dioctadecyldimethylammonium bromide, dioctadecyldimethylammonium chloride, 1,2-diacyl-3-trimethylammonium propane, 1,2-diacyl-3-dimethylammonium propane, [2,3-bis(oleoyl)propyl]trimethylammonium chloride or [N-(N-dimethylaminoethane)-carbamoyl]cholesterol, dioleoyl; dialkyl sulfosuccinates Salts, such as Aerosol OT; ethylenediaminetetrakis(ethoxylate-block-propoxylate) tetrols, such as Tetronic 304, Tetronic 904, Tetronic 90R4 or Tetronic 1304; fatty acids, such as palmitic acid, oleic acid, lauric acid, myristic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, palmitoleic acid, linoleic acid, linolenic acid or arachidonic acid and their salts, such as the sodium or potassium salts; glycosides, such as octyl glucoside or dodecyl maltoside; linear and branched alkyl benzene sulfonates; poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol), such as poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer 188 (Pluronic® F68 ), Poloxamer 212, Poloxamer 215, Poloxamer 217, Poloxamer 231, Poloxamer 234, Poloxamer 235, Poloxamer 237, Poloxamer 238, Poloxamer 282, Poloxamer 284, Poloxamer 288, Poloxamer 331, Poloxamer 333, Poloxamer 334, Poloxamer 335, Poloxamer 338, Poloxamer 401, Poloxamer 402, Poloxamer 403, Poloxamer 407, Poloxamer 105 benzoate or Poloxamer 182 dibenzoate;Polyoxyethylene sorbitan esters, such as polyethyleneoxy(40)-sorbitol hexaoleate, polyoxyethylene sorbitan monolaurate (Polysorbate 20, Tween® 20 and Tween® 21), polyoxyethylene sorbitan monopalmitate (Polysorbate 40, Tween® 40), polyoxyethylene sorbitan monostearate (Polysorbate 60, Tween® 60 and Tween® 61), polyoxyethylene sorbitan trioleate (Polysorbate 85, Tween® 85) or polyoxyethylene sorbitan tristearate (Polysorbate 65, Tween® 65); polyvinyl alcohol; polyvinylpyrrolidone; starch and derivatives thereof, and mixtures thereof.

[0588] In certain embodiments, the emulsifier is selected from the group consisting of sorbitan monolaurate (Span® 20), sorbitan monooleate (Span® 80), sorbitan monopalmitate (Span® 40), sorbitan monostearate (Span® 60), sorbitan sesquioleate (Span® 83), sorbitan trioleate (Span® 85), and sorbitan tristearate (Span® 65).

[0589] In certain embodiments, the emulsifier is selected from the group consisting of sorbitan monooleate (Span® 80), PEG-30 dipolyhydroxystearate (Cithrol™ DPHS), polyglyceryl-3 diisostearate, and a mixture of sorbitan oleate with a copolymer ester of hydroxystearic acid and ethylene glycol (Hypermer™ 1083).

[0590] In certain embodiments, the emulsifier is selected from the group consisting of sorbitan monooleate, PEG-30 dipolyhydroxystearate, polyglyceryl-3 diisostearate, and a mixture of sorbitan oleate with a copolymer ester of hydroxystearic acid and ethylene glycol.

[0591] In certain embodiments, the emulsifier is PEG-30 dipolyhydroxystearate. In certain embodiments, the emulsifier is polyglyceryl-3 diisostearate. In certain embodiments, the emulsifier is a mixture of sorbitan oleate and a copolymer ester of hydroxystearic acid and ethylene glycol.

[0592] Advantageously, the emulsifier is sorbitan monooleate.

[0593] The emulsifier may be added in an amount of about 0.01% (w / w) to about 15% (w / w). In certain embodiments, the emulsifier is added in an amount of about 0.01% (w / w) to about 10% (w / w). In certain embodiments, the emulsifier is added in an amount of about 0.1% (w / w) to about 7% (w / w). In certain embodiments, the emulsifier is added in an amount of about 1% (w / w) to about 5% (w / w). In certain embodiments, the emulsifier is added in an amount of about 1.5% (w / w) to about 3.0% (w / w).

[0594] In certain embodiments, the emulsifier is added in an amount of about 1.5% (w / w). In certain embodiments, the emulsifier is added in an amount of 1.5% (w / w). In certain embodiments, the emulsifier is added in an amount of about 0.5% (w / w). In certain embodiments, the emulsifier is added in an amount of 0.5% (w / w). In certain embodiments, the emulsifier is added in an amount of about 0.25% (w / w). In certain embodiments, the emulsifier is added in an amount of 0.25% (w / w).

[0595] In certain embodiments, the solvent can be any solvent that is immiscible with the dispersed phase.

[0596] In certain embodiments, the solvent is selected from the group consisting of polar solvents, non-polar solvents, fluorocarbons, and ionic liquids.

[0597] In certain embodiments, the solvent is a hydrocarbon, such as 3-carene, benzene, cumene, cycloheptane, cyclohexane, decane, dodecane, ethylbenzene, hemellitene, heptane, hexane, isodurene, limonene, mesitylene, m-xylene, n-butylbenzene, n-propylbenzene, nonane, octane, o-xylene, p-cymene, pentadecane, pentane, pinane, pinene, p-menthane, planite, pseudocumene, p- Selected from the group consisting of xylene, styrene, tetradecane, toluene, tridecane or undecane; siloxanes such as cyclomethicone, decamethylcyclopentasiloxane, hexamethyldisiloxane, octamethyltrisiloxane or liquid polysiloxanes (silicone oils), and esters such as acetyltributyl citrate, castor oil, ethyl laurate, glyceryl trioleate, liquid triglycerides, triacetin, tributyrin and triethyl citrate.

[0598] In certain embodiments, the solvent is selected from the group consisting of 3-carene, benzene, cumene, cycloheptane, cyclohexane, decane, dodecane, ethylbenzene, hemelitene, heptane, hexane, isodurene, limonene, mesitylene, m-xylene, n-butylbenzene, n-propylbenzene, nonane, octane, o-xylene, p-cymene, pentadecane, pentane, pinane, pinene, p-menthane, planitene, pseudocumene, p-xylene, styrene, tetradecane, toluene, tridecane, and undecane.

[0599] In certain embodiments, the solvent is selected from the group consisting of acetyl tributyl citrate, castor oil, ethyl laurate, glyceryl trioleate, liquid triglycerides, triacetin, tributyrin, and triethyl citrate.

[0600] In certain embodiments, the solvent is heptane or tetradecane. In certain embodiments, the solvent is heptane. In certain embodiments, the solvent is tetradecane.

[0601] In certain embodiments, solution B in step (a) comprises sorbitan monooleate and heptane. In certain embodiments, solution B in step (a) consists of sorbitan monooleate and heptane. In certain embodiments, solution B in step (a) comprises PEG-30 dipolyhydroxystearate and heptane.

[0602] In certain embodiments, solution B in step (a) comprises sorbitan monooleate and tetradecane. In certain embodiments, solution B in step (a) consists of sorbitan monooleate and tetradecane. In certain embodiments, solution B in step (a) comprises PEG-30 dipolyhydroxystearate and tetradecane.

[0603] In certain embodiments, solution B in step (a) comprises Hypermer™ 1083 and heptane. In certain embodiments, solution B in step (a) comprises a mixture of sorbitan oleate and a copolymer ester of hydroxystearic acid and ethylene glycol, and heptane.

[0604] In certain embodiments, solution B in step (a) comprises polyglyceryl-3 diisostearate and heptane.

[0605] Step (a) may be carried out at a temperature ranging from about 0°C to about 150°C, for example from about 4°C to about 80°C, for a time sufficient to allow the functionalized HA to react, for example from at least about 10 seconds to at least about 12 hours, for example from at least 30 minutes to at least about 12 hours.

[0606] In certain embodiments, the emulsion in step (a) is allowed to stand at room temperature for about 5 minutes. In certain embodiments, the emulsion in step (a) is allowed to stand at room temperature for about 12 hours. It is understood that room temperature can range from 17°C to 30°C, for example, from 17°C to 25°C.

[0607] Preferably, the pH adjuster initiates and / or accelerates the cross-linking reaction between the first functionalized HA and the second functionalized HA.

[0608] The pH adjuster may be soluble in both solution A and solution B of step (a), which provides the advantage of using controlled reaction conditions for the synthesis of the hydrogel HA microspheres of the present invention.

[0609] The pH adjuster may be an acid or a base.

[0610] In certain embodiments, the base is an aprotic, non-nucleophilic amine that is soluble in both the dispersed and continuous phases.

[0611] Exemplary bases are N,N,N',N'-tetramethylethylenediamine (TMEDA), 1,4-dimethylpiperazine, 4-methylmorpholine, 4-ethylmorpholine, 1,4-diazabicyclo[2.2.2]octane, 1,1,4,7,10,10-hexamethyltriethylenetetramine, 1,4,7-trimethyl-1,4,7-triazacyclononane, tris[2-(dimethylamino)ethyl]amine, triethylamine, and hexamethylenetetramine.

[0612] In certain embodiments, the base is selected from the group consisting of N,N,N',N'-tetramethylethylenediamine (TMEDA), 1,4-dimethylpiperazine, 4-methylmorpholine, 4-ethylmorpholine, 1,4-diazabicyclo[2.2.2]octane, 1,1,4,7,10,10-hexamethyltriethylenetetramine, 1,4,7-trimethyl-1,4,7-triazacyclononane, tris[2-(dimethylamino)ethyl]amine, 1,8-diazabicyclo[5.4.0]undec-7-ene, 1,5-diazabicyclo[4.3.0]non-5-ene, and hexamethylenetetramine. In certain embodiments, the base is N,N,N',N'-tetramethylethylenediamine (TMEDA).

[0613] In certain embodiments, prior to step (c), the suspension of step (b) is diluted with a solution containing a buffer, for example, a buffer having a pH of 4 or less. In certain embodiments, the solution contains succinic acid, sodium sulfate, and ethylenediaminetetraacetic acid (EDTA). In certain embodiments, the solution contains succinic acid, ethylenediaminetetraacetic acid (EDTA), and isopropanol. In certain embodiments, the solution contains sodium chloride.

[0614] In certain embodiments, in step (b), a pH adjuster is added to the emulsion of step (a), which is then incubated at a temperature ranging from 4 to 50° C., such as from 10 to 40° C., such as from 20 to 30° C. In certain embodiments, in step (b), the incubation is carried out at about 25° C., such as at 25° C.

[0615] In certain embodiments, in step (b), the pH adjuster increases the pH of the emulsion to about 4. In certain embodiments, in step (a), the pH of the emulsion, i.e., the pH before addition of the pH adjuster, is at least 1, for example about 2. It is understood that in step (a), the pH of the emulsion is measured in the aqueous phase.

[0616] Suitably, in step (d), size fractionation may be carried out by sieving, such as by wet sieving, for example by using a vibrating sieve separator, stirred cell filtration, cross-flow filtration, sedimentation, or centrifugation.

[0617] Advantageously, size fractionation is carried out by wet sieving.

[0618] In certain embodiments, in step (d), the obtained hydrogel HA microspheres are wet sieved in a solvent that allows the dispersed phase particles to swell.

[0619] In certain embodiments, in step (d), the obtained hydrogel HA microspheres are wet-sieved in a solvent, such as a solvent containing a buffer and optionally a water-miscible organic solvent, such as a polar solvent. Exemplary solvents may be selected from the group consisting of ethanol, methanol, isopropanol, acetonitrile, dioxane, dimethylformamide, dimethyl sulfoxide, tert-butanol, dimethylacetamide, and N-methylpyrrolidone.

[0620] In certain embodiments, in step (e), the microspheres are washed with a solution containing a buffer, such as succinic acid. In certain embodiments, the solution contains succinic acid, sodium chloride, ethylenediaminetetraacetic acid, and polyoxyethylenesorbitan monolaurate. In certain embodiments, in step (e), the microspheres are washed with a solution containing succinic acid, sodium chloride, histidine, and poloxamer (Pluronic™ F-68).

[0621] Chemical modification of HA with functional groups confers functionality to the HA. Those skilled in the art will recognize that a single functionalized HA may have more than one reactive functional group, e.g., -FG1 or -FG2.

[0622] In certain embodiments, -FG1, -FG2, and -FG3 are

[0623] [ka] TIFF2026501742000110.tif220163TIFF2026501742000111.tif220163TIFF2026501742000112.tif131163, wherein the dashed line indicates the bond to the first or second functionalized HA, for example to the variable -X'- or -Y'-, Each-R 08 , -R 08a and -R 08b are halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 09 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 010 )-, -S(O)2N(R 010 )-, -S(O)N(R 010 )-, -S(O)2-, -S(O)-, -N(R 010 )S(O)2N(R 010a )-, -S-, -N(R 010 )-, -OC(OR 010 )(R 010a )-, -N(R 010 )C(O)N(R 010a )- and -OC(O)N(R 010 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 09 and optionally independently substituted with Each-R 09 , -R 010 and -R 010a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each -Y 01 are independently selected from the group consisting of -F, -Cl, -Br, and -I; each n is independently 1, 2, 3, or 4; Each -Y 02 and -Y 02a are independently selected from the group consisting of -H and -Br; Each -Y 03 and -Y 03a -F, -Cl, -Br, -I, -OR, -NR 011 R 011a and -SR 011 are independently selected from the group consisting of Each -Y 04 -, -O-, -S-, -NR 011 -, -CR 011 R 011a - selected independently from Each-R 011 and -R 011a are halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 012 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 013 )-, -S(O)2N(R 013 )-, -S(O)N(R 013 )-, -S(O)2-, -S(O)-, -N(R 013 )S(O)2N(R 013a )-, -S-, -N(R 013 )-, -OC(OR 013 )(R 013a )-, -N(R 013 )C(O)N(R 013a )- and -OC(O)N(R 013 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 012 and optionally independently substituted with Each-R 12 , -R 013 and -R 013a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. are independently selected from the group consisting of:

[0624] In certain embodiments, -FG1, -FG2, and -FG3 are

[0625] [ka] TIFF2026501742000114.tif137163, wherein the dashed line indicates the bond to the first or second functionalized HA, for example to the variable -X'- or -Y'-, Each-R 08 , -R 08a and -R 08bare halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 09 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 010 )-, -S(O)2N(R 010 )-, -S(O)N(R 010 )-, -S(O)2-, -S(O)-, -N(R 010 )S(O)2N(R 010a )-, -S-, -N(R 010 )-, -OC(OR 010 )(R 010a )-, -N(R 010 )C(O)N(R 010a )- and -OC(O)N(R 010 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 09 and optionally independently substituted with Each-R 09 , -R 010 and -R 010a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each -Y 01are independently selected from the group consisting of -F, -Cl, -Br, and -I; each n is independently 1, 2, 3, or 4; Each -Y 02 and -Y 02a are independently selected from the group consisting of -H and -Br; Each -Y 03 and -Y 03a -F, -Cl, -Br, -I, -OR, -NR 011 R 011a and -SR 011 are independently selected from the group consisting of Each -Y 04 -, -O-, -S-, -NR 011 -, -CR 011 R 011a - selected independently from Each-R 011 and -R 011a are halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 012 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 013 )-, -S(O)2N(R 013 )-, -S(O)N(R 013 )-, -S(O)2-, -S(O)-, -N(R 013 )S(O)2N(R 013a )-, -S-, -N(R 013 )-, -OC(OR 013 )(R 013a )-, -N(R 013 )C(O)N(R 013a )- and -OC(O)N(R 013)-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 012 and optionally independently substituted with Each-R 12 , -R 013 and -R 013a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. are independently selected from the group consisting of:

[0626] In certain embodiments, the pH adjuster increases the pH of the emulsion of step (a), and -FG1, -FG2, and -FG3 are

[0627] [ka] TIFF2026501742000116.tif23163, wherein the dashed line indicates the bond to the first or second functionalized HA, for example to the variable -X'- or -Y'-, Each-R 08 and -R 08a are halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 09 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 010 )-, -S(O)2N(R 010 )-, -S(O)N(R 010 )-, -S(O)2-, -S(O)-, -N(R 010 )S(O)2N(R 010a )-, -S-, -N(R 010 )-, -OC(OR 010 )(R 010a )-, -N(R 010 )C(O)N(R 010a )- and -OC(O)N(R 010 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 09 and optionally independently substituted with Each-R 09 , -R 010 and -R 010a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each -Y 01 are independently selected from the group consisting of -F, -Cl, -Br, and -I; each n is independently 1, 2, 3, or 4; Each -Y 02 and -Y 02a are independently selected from the group consisting of -H and -Br; Each -Y 03 and -Y 03a -F, -Cl, -Br, -I, -OR, -NR 011 R 011a and -SR 011 are independently selected from the group consisting of Each-R011 and -R 011a are halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 012 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 013 )-, -S(O)2N(R 013 )-, -S(O)N(R 013 )-, -S(O)2-, -S(O)-, -N(R 013 )S(O)2N(R 013a )-, -S-, -N(R 013 )-, -OC(OR 013 )(R 013a )-, -N(R 013 )C(O)N(R 013a )- and -OC(O)N(R 013 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 012 and optionally independently substituted with Each-R 012 , -R 013 and -R 013a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. are independently selected from the group consisting of:

[0628] In certain embodiments, the pH adjuster decreases the pH of the emulsion of step (a), and -FG1 and -FG2 are

[0629] [ka] wherein the dashed line indicates the bond to the first or second functionalized HA, for example to the variable -X'- or -Y'-, Each-R 08 and -R 08a are halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 09 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 010 )-, -S(O)2N(R 010 )-, -S(O)N(R 010 )-, -S(O)2-, -S(O)-, -N(R 010 )S(O)2N(R 010a )-, -S-, -N(R 010 )-, -OC(OR 010 )(R 010a )-, -N(R 010 )C(O)N(R 010a )- and -OC(O)N(R 010 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 09 and optionally independently substituted with Each-R 09 , -R 010 and -R 010a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. are independently selected from the group consisting of:

[0630] In certain embodiments, -FG1 is

[0631] [ka] wherein the dashed line indicates the bond to the first functionalized HA, e.g., to the variable -X'-; -Y 01 are independently selected from the group consisting of -F, -Cl, -Br, and -I; Each-R 08 and -R 08a are halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 09 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 010 )-, -S(O)2N(R 010 )-, -S(O)N(R 010)-, -S(O)2-, -S(O)-, -N(R 010 )S(O)2N(R 010a )-, -S-, -N(R 010 )-, -OC(OR 010 )(R 010a )-, -N(R 010 )C(O)N(R 010a )- and -OC(O)N(R 010 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 09 and optionally independently substituted with Each-R 09 , -R 010 and -R 010a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. are independently selected from the group consisting of -FG2 is

[0632] [ka] wherein the dashed line indicates the bond to the second functionalized HA, e.g., to the variable -Y'-; Each -Y 02 and -Y 02a are independently selected from the group consisting of -H and -Br. are independently selected from the group consisting of provided that when -FG1 is of formula (y-56), -FG2 is of formula (y-57) or (y-86); when -FG1 is of formula (y-1), -FG2 is of formula (y-16) or (y-47); when -FG1 is of formula (y-44), -FG2 is of formula (y-16) or (y-47); when -FG1 is of formula (y-6), -FG2 is of formula (y-9); when -FG1 is of formula (y-49), -FG2 is of formula (y-85); when -FG1 is of formula (y-44), -FG2 is of formula (y-47); or when -FG1 is of formula (y-39), -FG2 is of formula (y-56).

[0633] In certain embodiments, the pH adjuster increases the pH of the emulsion of step (a), and -FG1

[0634] [ka] wherein the dashed line indicates the bond to the first functionalized HA, e.g., to the variable -X'-; -Y 01 are independently selected from the group consisting of -F, -Cl, -Br, and -I; Each-R 08 and -R 08a are halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 09 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 010)-, -S(O)2N(R 010 )-, -S(O)N(R 010 )-, -S(O)2-, -S(O)-, -N(R 010 )S(O)2N(R 010a )-, -S-, -N(R 010 )-, -OC(OR 010 )(R 010a )-, -N(R 010 )C(O)N(R 010a )- and -OC(O)N(R 010 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 09 and optionally independently substituted with Each-R 09 , -R 010 and -R 010a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. are independently selected from the group consisting of -FG2 is

[0635] [ka] wherein the dashed line indicates the bond to the second functionalized HA, e.g., to the variable -Y'-; Each -Y 02 and -Y 02a are independently selected from the group consisting of -H and -Br. are independently selected from the group consisting of provided that when -FG1 is of formula (y-56), -FG2 is of formula (y-57) or (y-86); when -FG1 is of formula (y-1), -FG2 is of formula (y-16); when -FG1 is of formula (y-44), -FG2 is of formula (y-16); or when -FG1 is of formula (y-39), -FG2 is of formula (y-56).

[0636] In certain embodiments, the pH adjuster decreases the pH of the emulsion of step (a), and -FG1

[0637] [ka] wherein the dashed line indicates the bond to the first functionalized HA, e.g., to the variable -X'-; Each-R 08 , -R 08a are halogens, -H, -CN, -T 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 alkynyl; 0 , C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl may be one or more -R 09 is optionally replaced by C 1~50 Alkyl, C 2~50 Alkenyl, and C 2~50 Alkynyl is -T 0 -, -C(O)O-, -O-, -C(O)-, -C(O)N(R 010 )-, -S(O)2N(R 010 )-, -S(O)N(R 010 )-, -S(O)2-, -S(O)-, -N(R 010 )S(O)2N(R 010a )-, -S-, -N(R 010 )-, -OC(OR 010 )(R 010a )-, -N(R 010 )C(O)N(R010a )- and -OC(O)N(R 010 )-, Each T 0 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3~10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; 0 is one or more -R 09 and optionally independently substituted with Each-R 09 , -R 010 and -R 010a -H and C 1~6 alkyl, independently selected from the group consisting of C 1~6 The alkyl is optionally substituted with one or more halogens which may be the same or different. are independently selected from the group consisting of -FG2 is

[0638] [ka] where the dashed line indicates the bond to the second functionalized HA, e.g., to the variable -Y'-. are independently selected from the group consisting of However, when -FG1 is of formula (y-6), -FG2 is of formula (y-9), and when -FG1 is of formula (y-44), -FG2 is of formula (y-47).

[0639] In certain embodiments, -FG1 is

[0640] [ka] where the dashed line indicates the bond to the first functionalized HA, e.g., to the variable -X'-. and -FG2 is

[0641] [ka] where the dashed line indicates the bond to the second functionalized HA, e.g., to the variable -Y'-. and the pH adjuster increases the pH of the emulsion of step (a) from about 1 to about 9, such as from about 1 to about 5.5, or such as from about 2 to about 4, and each -Y 02 and -Y 02a is independently selected from the group consisting of -H and -Br.

[0642] In certain embodiments, -FG1 is

[0643] [ka] where the dashed line indicates the bond to the first functionalized HA, e.g., to the variable -X'-. and -FG2 is

[0644] [ka] where the dashed line indicates the bond to the second functionalized HA, e.g., to the variable -Y'-. and the pH adjuster increases the pH of the emulsion of step (a) from about 1 to about 9, such as from about 1 to about 5.5, or such as from about 2 to about 4; -Y 02 and -Y 02a Both are -H.

[0645] In certain embodiments, -FG1 is

[0646] [ka] where the dashed line indicates the bond to the first functionalized HA, e.g., to the variable -X'-. and -FG2 is

[0647] [ka] where the dashed line indicates the bond to the second functionalized HA, e.g., to the variable -Y'-. and the pH adjuster increases the pH of the emulsion of step (a) from 1 to 5.5, and each -Y 02 and -Y 02a is independently selected from the group consisting of -H and -Br.

[0648] In certain embodiments, -FG1 is

[0649] [ka] where the dashed line indicates the bond to the first functionalized HA, e.g., to the variable -X'-. and -FG2 is

[0650] [ka] where the dashed line indicates the bond to the second functionalized HA, e.g., to the variable -Y'. and the pH adjuster increases the pH of the emulsion of step (a) from 1 to 5.5, and -Y 02 and -Y 02a Both are -H.

[0651] In certain embodiments, -FG1 is

[0652] [ka] where the dashed line indicates the bond to the first functionalized HA, e.g., to the variable -X'-. and -FG2 is

[0653] [ka] where the dashed line indicates the bond to the second functionalized HA, e.g., to the variable -Y'-. and the pH adjuster increases the pH of the emulsion of step (a) from 2 to 4, and each -Y 02 and -Y 02a is independently selected from the group consisting of -H and -Br.

[0654] In certain embodiments, -FG1 is

[0655] [ka] where the dashed line indicates the bond to the first functionalized HA, e.g., to the variable -X'-. and -FG2 is

[0656] [ka] where the dashed line indicates the bond to the second functionalized HA, e.g., to the variable -Y'-. and the pH adjuster increases the pH of the emulsion of step (a) from 2 to 4, and -Y 02 and -Y 02a Both are -H.

[0657] In certain embodiments, -FG3 has formula (y-56) or (y-57):

[0658] [ka] [Wherein the formula, the dashed line represents -L 2 - indicates a bond to -Y 02 and -Y 02a are independently selected from the group consisting of -H and -Br. Either:

[0659] Preferably, -FG3 is

[0660] [ka] [Wherein the formula, the dashed line represents -L2 - indicates a bond to is.

[0661] In certain embodiments, the method of the present invention comprises: (a) mixing solution A with solution B to form an emulsion, wherein solution A comprises a first functionalized HA modified with one or more -FG1 and optionally further functional groups, and a second functionalized HA modified with one or more -FG2 and optionally further functional groups, -FG1 and -FG2 being different functional moieties, and -FG1 on the first functionalized HA reacts with -FG2 on the second functionalized HA to form multiple crosslinks resulting in the formation of hydrogel HA microspheres; The first functionalized HA is linearly connected to the following Z 1 Units and Z 5 Each of the units:

[0662] [ka] Contains multiple The second functionalized HA is linearly connected to the following Z 1 Units and Z 6 Each of the units:

[0663] [ka] Contains multiple During the ceremony, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each R a1 -H, C 1~10 independently selected from the group consisting of alkyl, ammonium ion, tetrabutylammonium ion, cetyltrimethylammonium ion, alkali metal ion, and alkaline earth metal ion; Each-R a2are independently -H or C 1~10 is alkyl, each -FG1, -FG2 is defined as elsewhere herein; each -X- and -Y- is independently a carbonyl group or is absent; each -X'-, -Y'- is independently a spacer moiety or is absent; (b) optionally adding a pH adjuster to the emulsion of step (a); (c) collecting the resulting hydrogel HA microspheres of step (a) or (b), wherein the hydrogel is 3 unit:

[0664] [ka] [In the formula, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each-R a2 , -X-, -Y-, -X'- and -Y'- are defined as in step (a); Each-L 3 - is independently a linking moiety or is absent] including multiple steps, (d) optionally, size fractionating the resulting hydrogel HA microspheres of step (a), (b), or (c) to obtain microspheres having a particular particle size distribution; (e) optionally washing the hydrogel HA microspheres obtained in step (a), (b), (c) or (d); (f) optionally, collecting the hydrogel HA microspheres of step (d) or (e); (g) providing the hydrogel HA microspheres or pharmaceutically acceptable salts thereof of step (c) or (f), wherein the hydrogel contains one o...

Claims

1. 1. A drug conjugate, or a pharmaceutically acceptable salt thereof, comprising a hyaluronic acid (HA) hydrogel microsphere, or a pharmaceutically acceptable salt thereof, comprising crosslinked HA chains to which a plurality of drug moieties are covalently and reversibly conjugated, said drug conjugate comprising each of the following units: 【Chemistry 1】 [In the formula, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each R a1 -H, C 1～10 independently selected from the group consisting of alkyl, ammonium ion, tetrabutylammonium ion, cetyltrimethylammonium ion, alkali metal ion, and alkaline earth metal ion; Each-R a2 are independently -H or C 1～10 is alkyl, each -X- and -Y- is independently a carbonyl group or is absent; each -X'-, -Y'- is independently a spacer moiety or absent; Each -D can independently 1 a VEGF-neutralizing drug moiety covalently and reversibly conjugated to Each-L 1 - is independently a reversible linker moiety; Each-L 2 - is independently a spacer moiety or absent; Each-L 3 -, -L 4 -, -L 5 - is independently a linking moiety or absent; each -BA is independently a blocking agent] A drug conjugate or a pharmaceutically acceptable salt thereof, comprising a plurality of

2. The drug conjugate has a Z in the range of about 50% to about 98% 1 , Z in the range of about 0.1% to about 20% 2 , Z in the range of about 0.1% to about 20% 3 and Z in the range of about 0.1% to about 10% 4 2. The drug conjugate of claim 1, or a pharmaceutically acceptable salt thereof, comprising:

3. The drug conjugate has a Z in the range of about 75% to about 98% 1 , Z in the range of about 0.1% to about 10% 2 , Z in the range of about 0.1% to about 10% 3 and Z in the range of about 0.1% to about 5% 4 3. The drug conjugate of claim 1 or 2, or a pharmaceutically acceptable salt thereof, comprising:

4. The drug conjugate has a Z in the range of about 78% to about 96% 1 , Z in the range of about 2% to about 10% 2 , Z in the range of about 1% to about 7% 3 and Z in the range of about 0.5% to about 5% 4 The drug conjugate according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, comprising:

5. 5. The drug conjugate or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein each -D is a ranibizumab moiety.

6. -BA, 【Chemistry 2】 [Wherein the formula, the dashed line represents -L 5 - indicates a bond to The drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, which is selected from the group consisting of:

7. -L 3 -, -L 4 - and -L 5 - is the formula (y): 【Transformation 3】 [In the formula, -L 3 For -, the dashed line with an asterisk indicates the bond to -Y'-, the unmarked dashed line indicates the bond to -X'-, and -L 4 -, the dashed line with an asterisk indicates the bond to -Y'-, and the unmarked dashed line indicates the bond to -L 2 - indicates a bond to -L 5 -, the dashed line with an asterisk indicates the bond to -Y'-, and the unmarked dashed line indicates the bond to -BA] The drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6,

8. The drug conjugate comprises each of the following units: 【Chemistry 4】 [In the formula, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each R a1 -H, C 1～10 independently selected from the group consisting of alkyl, ammonium ion, tetrabutylammonium ion, cetyltrimethylammonium ion, alkali metal ion, and alkaline earth metal ion; Each-R a2 are independently -H or C 1～10 is alkyl, each -X'-, -Y'- is independently a spacer moiety or absent; Each -D is a ranibizumab moiety, Each-L 1 - is independently a reversible linker moiety; Each-L 2 - is independently a spacer moiety or absent] The drug conjugate or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, comprising a plurality of

9. Each R a1 is H or an alkali metal ion, -R a2 The drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, wherein is -H.

10. The drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, wherein each of -X- and -Y- is a carbonyl group.

11. Each of -X'- and -Y'- is independently -T'-, C 1～50 Alkyl, C 2～50 Alkenyl, and C 2～50 alkynyl, and C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 The alkynyl may be one or more of the same or different -R y1 is optionally replaced by C 1～50 Alkyl, C 2～50 Alkenyl, and C 2～50 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y2 )-, -S(O) 2 N(R y2 )-, -S(O)N(R y2 )-, -S(O) 2 -, -S(O)-, -N(R y2 )S(O) 2 N(R y2a )-, -S-, -N(R y2 )-, -OC(OR y2 )(R y2a )-, -N(R y2 )C(O)N(R y2a )- and -OC(O)N(R y2 )-, Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3～10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; and each T' is the same or different and is selected from the group consisting of one or more -R y1 and optionally independently substituted with Each-R y1 But halogen, -CN, oxo(=O), -COOR y3 , -OR y3 , -C(O)R y3 , -C(O)N(R y3 R y3a ), -S(O) 2 N(R y3 R y3a ), -S(O)N(R y3 R y3a ), -S(O) 2 R y3 , -S(O)R y3 , -N(R y3 )S(O) 2 N(R y3a R y3b ), -SR y3 , -N(R y3 R y3a ), -NO 2 , -OC(O)R y3 , -N(R y3 )C(O)R y3a , -N(R y3 )S(O) 2 R y3a , -N(R y3 )S(O)R y3a , -N(R y3 )C(O)OR y3a , -N(R y3 )C(O)N(R y3a R y3b ), -OC(O)N(R y3 R y3a ), and C 1～6 alkyl, independently selected from the group consisting of C 1～6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R y2 , -R y2a , -R y3 , -R y3a , -R y3b But -H and C 1～6 alkyl, independently selected from the group consisting of C 1～6 11. The drug conjugate of any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein the alkyl is optionally substituted with one or more halogens which are the same or different.

12. Each -X'- has the formula (x0): 【Transformation 5】 [In the formula, The unmarked dashed line indicates a bond to -X-, and the dashed line with an asterisk indicates a bond to -L 3 - indicates a bond to v 0 is selected from the group consisting of 0 and 1; -X 1 -, -X 4 - is independently C 1～10 This C is an alkyl 1～10 Alkyl is -O-, -T-, -N(R y1 )-, -C(O)O- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1～10 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a Optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -R y2a -H and C 1～4 independently selected from the group consisting of alkyl, -X 2 - is -N(R 1 )—, —O—, —S—, and —Se—; =X 3 is =O, =N(R 1 ) and =S; -X 5 -C 1～20 This C is an alkyl 1～20 Alkyl is -O-, -C(O)O-, -T-, -N(R y1 )- and -N(R y1 )C(O)—, wherein the C 1～20 The alkyl chain is -OH, -T, -N(R y1 )- and -C(O)N(R y2 R y2a Optionally substituted with one or more groups independently selected from -R 1 -H, C 1～5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3～10 and each T is independently selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from one or more -R 2 and optionally independently substituted with -R 2 is halogen, -CN, oxo, -C(O)OR 3 , -OR 3 , -C(O)R 3 , -C(O)N(R 3 )(R 3a ), -S(O) 2 N(R 3 )(R 3a ), -S(O)N(R 3 )(R 3a ), -S(O) 2 R 3 , -S(O)R 3 , -N(R 3 )S(O) 2 N(R 3a )(R 3b ), -SR 3 , -N(R 3 )(R 3a ), -NO 2 , -OC(O)R 3 , -N(R 3 )C(O)R 3a , -N(R 3 )S(O) 2 R 3a , -N(R 3 )S(O)R 3a , -N(R 3 )C(O)OR 3a , -N(R 3 )C(O)N(R 3a )(R 3b ), -OC(O)N(R 3 )(R 3a ) and C 1～6 alkyl, C 1～6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 3 , -R 3a and -R 3b -H and C 1～6 alkyl, independently selected from the group consisting of C 1～6 The alkyl is optionally substituted with one or more halogens which may be the same or different. The drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11,

13. Each -Y'- has the formula (y0): 【Transformation 6】 [In the formula, The unmarked dashed line indicates a bond to -Y-, and the dashed line with an asterisk indicates a bond to -L 3 -, -L 4 -or-L 5 - indicates a bond to -Y 1 -, -Y 4 - is independently C 1～10 This C is an alkyl 1～10 Alkyl is -O-, -T-, -N(R y1 )-, -C(O)O- and -C(O)N(R y1 )-, wherein C is optionally interrupted by one or more groups independently selected from 1～10 The alkyl chain is -OH, -T, -NH(R y1 ) and -C(O)N(R y2 R y2a ) optionally substituted with one or more groups independently selected from -R y1 , -R y2 , -R y2a is H and C 1～4 independently selected from the group consisting of alkyl, -Y 2 - is -N(R 2 )—, —O—, —S—, and —Se—; =Y 3 is =O, =N(R 2 ) and =S; -R 1 , -R 2 -H, C 1～5 independently selected from the group consisting of alkyl and -T; Each T is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3～10 and each T is independently selected from the group consisting of cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl, and each T is the same or different, and is selected from one or more -R 3 and optionally independently substituted with -R 3 is halogen, -CN, oxo, -C(O)OR 4 , -OR 4 , -C(O)R 4 , -C(O)N(R 4 )(R 4a ), -S(O) 2 N(R 4 )(R 4a ), -S(O)N(R 4 )(R 4a ), -S(O) 2 R 4 , -S(O)R 4 , -N(R 4 )S(O) 2 N(R 4a )(R 4b ), -SR 4 , -N(R 4 )(R 4a ), -NO 2 , -OC(O)R 4 , -N(R 4 )C(O)R 4a , -N(R 4 )S(O) 2 R 4a , -N(R 4 )S(O)R 4a , -N(R 4 )C(O)OR 4a , -N(R 4 )C(O)N(R 4a )(R 4b ), -OC(O)N(R 4 )(R 4a ) and C 1～6 alkyl, C 1～6 the alkyl is optionally substituted with one or more halogens which may be the same or different; -R 4 , -R 4a and -R 4b -H and C 1～6 alkyl, independently selected from the group consisting of C 1～6 The alkyl is optionally substituted with one or more halogens which may be the same or different. The drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein

14. Each-L 1 is of formula (I): 【Transformation 7】 [In the formula, The dashed line indicates the attachment of -D to the nitrogen atom by forming an amide bond; -X- is -C(R 4 R 4a )-, -N(R 4 )-, -O-, -C(R 4 R 4a )-C(R 5 R 5a )-, -C(R 5 R 5a )-C(R 4 R 4a )-, -C(R 4 R 4a )-N(R 6 )-, -N(R 6 )-C(R 4 R 4a )-, -C(R 4 R 4a )-O-, -OC(R 4 R 4a )- or -C(R 7 R 7a )- and X 1 is C or S(O), -X 2 - is -C(R 8 R 8a )- or -C(R 8 R 8a )-C(R 9 R 9a )- and =X 3 is =O, =S, or =N-CN, -R 1 , -R 1a , -R 2 , -R 2a , -R 4 , -R 4a , -R 5 , -R 5a , -R 6 , -R 8 , -R 8a , -R 9 , -R 9a -H, and C 1～6 independently selected from the group consisting of alkyl, -R 3 , -R 3a -H, and C 1～6 alkyl, with the proviso that -R 3 , -R 3a If one or both of are other than -H, they are sp 3 provided that it is connected to a bonded N through a hybridized carbon atom, -R 7 is -N(R 10 R 10a ), or -NR 10 -(C=O)-R 11 and -R 7a , -R 10 , -R 10a , -R 11 are, independently of each other, -H or C 1～10 is alkyl, Optionally, Pair-R 1a / -R 4a , -R 1a / -R 5a , -R 1a / -R 7a , -R 4a / -R 5a , -R 8a / -R 9a one or more of the following forms a chemical bond; Optionally, Pair-R 1 / -R 1a , -R 2 / -R 2a , -R 4 / -R 4a , -R 5 / -R 5a , -R 8 / -R 8a , -R 9 / -R 9a One or more of the following, together with the atoms to which they are attached, form a C 3～10 forming a cycloalkyl or a 3- to 10-membered heterocyclyl; Optionally, Pair-R 1 / -R 4 , -R 1 / -R 5 , -R 1 / -R 6 , -R 1 / -R 7a , -R 4 / -R 5 , -R 4 / -R 6 , -R 8 / -R 9 , -R 2 / -R 3 together with the atoms to which they are attached form ring A, Optionally, -R 3 / -R 3a together with the nitrogen atom to which they are attached form a 3- to 10-membered heterocyclic ring, Ring A is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3～10 cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl. It is of Each-L 1 -Ga-L 2 - and optionally further substituted, with the proviso that the hydrogen marked with an asterisk in formula (I) is not replaced by a substituent, or a pharmaceutically acceptable salt thereof.

15. Each-L 1 15. The drug conjugate of claim 14, or a pharmaceutically acceptable salt thereof, wherein - is not optionally further substituted.

16. -X- is -C(R 7 R 7a )- and -R 7 Ga-NR 10 -(C=O)-R 11 and -R 7a The drug conjugate or a pharmaceutically acceptable salt thereof according to claim 14 or 15, wherein is -H.

17. Each-L 2 - is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y1 )-, -S(O) 2 N(R y1 )-, -S(O)N(R y1 )-, -S(O) 2 -, -S(O)-, -N(R y1 )S(O) 2 N(R y1a )-, -S-, -N(R y1 )-, -OC(OR y1 )(R y1a )-, -N(R y1 )C(O)N(R y1a )-, -OC(O)N(R y1 )-, C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 alkynyl, -T'-, C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 The alkynyl may be one or more of the same or different -R y2 is optionally replaced by C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y3 )-, -S(O) 2 N(R y3 )-, -S(O)N(R y3 )-, -S(O) 2 -, -S(O)-, -N(R y3 )S(O) 2 N(R y3a )-, -S-, -N(R y3 )-, -OC(OR y3 )(R y3a )-, -N(R y3 )C(O)N(R y3a )- and -OC(O)N(R y3 )-, -R y1 and -R y1a But, -H, -T', C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 alkynyl, -T', C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 The alkynyl may be one or more of the same or different -R y2 is optionally replaced by C 1～50 Alkyl, C 2～50 Alkenyl and C 2～50 Alkynyl is -T'-, -C(O)O-, -O-, -C(O)-, -C(O)N(R y4 )-, -S(O) 2 N(R y4 )-, -S(O)N(R y4 )-, -S(O) 2 -, -S(O)-, -N(R y4 )S(O) 2 N(R y4a )-, -S-, -N(R y4 )-, -OC(OR y4 )(R y4a )-, -N(R y4 )C(O)N(R y4a )- and -OC(O)N(R y4 )-, Each T' is phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3～10 cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; and each T' is the same or different and is selected from the group consisting of one or more -R y2 and optionally independently substituted with Each-R y2 But halogen, -CN, oxo(=O), -C(O)OR y5 , -OR y5 , -C(O)R y5 , -C(O)N(R y5 )(R y5a ), -S(O) 2 N(R y5 )(R y5a ), -S(O)N(R y5 )(R y5a ), -S(O) 2 R y5 , -S(O)R y5 , -N(R y5 )S(O) 2 N(R y5 )(R y5a ), -SR y5 , -N(R y5 )(R y5a ), -NO 2 , -OC(O)R y5 , -N(R y5 )C(O)R y5a , -N(R y5 )S(O) 2 R y5a , -N(R y5 )S(O)R y5a , -N(R y5 )C(O)OR y5a , -N(R y5 )C(O)N(R y5 )(R y5a ), -OC(O)N(R y5 )(R y5a ), and C 1～6 alkyl, independently selected from the group consisting of C 1～6 the alkyl is optionally substituted with one or more halogens which may be the same or different; Each-R y3 , -R y3a , -R y4 , -R y4a , -R y5 , -R y5a and -R y5b But -H and C 1～6 alkyl, independently selected from the group consisting of C 1～6 17. The drug conjugate of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, wherein the alkyl is optionally substituted with one or more halogens which are the same or different.

18. Each-L 2 -L 1 - is the formula (s1): 【Transformation 8】 [Wherein the formula, the dashed line represents -L 4 The dashed line with an asterisk indicates the bond to the ranibizumab moiety, -D. The drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 17,

19. The drug conjugate comprises each of the following units: 【Chemistry 9】 【change】 [In the formula, An unmarked dashed line indicates a point of attachment to the adjacent unit or to a hydrogen atom at the dashed line marked with a #; The dashed lines marked with # indicate the point of attachment to the adjacent unit or to the hydroxyl group at the unmarked dashed line; Each R a1 is H or an alkali metal ion, Each-R a2 is -H, Each -D is a ranibizumab moiety, Each -X'- has the formula (x4): 【Chemistry 10】 (wherein the unmarked dashed line indicates the bond to the carbonyl group, the dashed line marked with an asterisk indicates the bond to the sulfur atom, and c 0 is 7) It is of Each -Y'- has the formula (y4): 【Chemistry 11】 (wherein the unmarked dashed line indicates the bond to the carbonyl group and the dashed line marked with an asterisk indicates the bond to the nitrogen atom of the thiosuccinimide ring). It is of Each-L 2 -L 1 - is the formula (s1) or (s2): 【Chemistry 12】 where the dashed line indicates the bond to the sulfur atom and the dashed line with an asterisk indicates the bond to the ranibizumab moiety, -D. [It is of the] The drug conjugate or pharmaceutically acceptable salt thereof according to any one of claims 1 to 18, comprising a plurality of

20. Each-L 2 -L 1 is of formula (s2) or (s1), and the drug conjugate has a Z of about 92.9% 1 , about 4.3% Z 2 -i, about 1.5% Z 3 -i and approximately 1.3% Z 4 20. The drug conjugate of claim 19, or a pharmaceutically acceptable salt thereof, comprising -i.

21. Each -L 2 -L 1 -が、Formula (s2): 【Chemistry 13】 where the dashed line indicates the bond to the sulfur atom and the dashed line with an asterisk indicates the bond to the ranibizumab moiety, -D. and the drug conjugate has a Z of 92.9%. 1 , 4.3% Z 2 -i, 1.5% Z 3 -i and 1.3% Z 4 21. The drug conjugate or a pharmaceutically acceptable salt thereof of claim 19 or 20, comprising -i.

22. A pharmaceutical composition comprising the drug conjugate of any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

23. The drug conjugate according to any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 22, for use as a pharmaceutical.

24. The drug conjugate of any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 22, for use in reducing or inhibiting angiogenesis in a subject having a disorder associated with pathological angiogenesis.

25. The drug conjugate of any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 22, for use in treating a disorder associated with pathological angiogenesis.

26. The drug conjugate or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition for use according to any one of claims 23 to 25, wherein the disorder associated with pathological angiogenesis is an eye disorder.

27. The ocular disorder is age-related macular degeneration (AMD), macular degeneration, macular edema, diabetic macular edema (DME), retinopathy, diabetic retinopathy (DR), other ischemia-related retinopathies, retinopathy of prematurity (ROP), retinal vein occlusion (RVO), CNV, corneal neovascularization, diseases related to corneal neovascularization, retinal neovascularization, uveitis macular edema, branch retinal vein occlusion (BRVO), central retinal vein occlusion (CRVO), submacular hemorrhage, polypoidal choroidal vasculopathy (PCV), retinal capillary microaneurysm, retinal artery occlusion (RAO), branch retinal artery occlusion (BRAO), central retinal artery occlusion (CRAO), subfoveal hemorrhage, subretinal hemorrhage, radiation retinopathy, exudative retinal detachment, Eales' disease, neovascular macular telangiectasia, ischemic retinal vasculitis, diseases related to retinal or choroidal neovascularization 27. The drug conjugate or pharmaceutically acceptable salt thereof, or pharmaceutical composition for use according to claim 26, wherein the drug conjugate or pharmaceutically acceptable salt thereof is selected from the group consisting of ocular histoplasmosis, pathologic myopia, von Hippel-Lindau disease, ocular histoplasmosis, familial exudative vitreoretinopathy (FEVR), Coats' disease, Norrie's disease, osteoporosis-pseudoglioma syndrome (OPPG), subconjunctival hemorrhage, rubeosis, ocular neovascular diseases, neovascular glaucoma, retinitis pigmentosa (RP), hypertensive retinopathy, retinal angiomatous proliferation, macular telangiectasia, iris neovascularization, intraocular neovascularization, retinal degeneration, cystoid macular edema (CME), vasculitis, papilledema, retinitis, conjunctivitis, Leber's congenital amaurosis, uveitis, choroiditis, ocular histoplasmosis, blepharitis, dry eye, traumatic eye injury, and Sjogren's disease.

28. Eye disorders include age-related macular degeneration (AMD), macular degeneration, macular edema, diabetic macular edema (DME), retinopathy, diabetic retinopathy (DR), other ischemia-related retinopathies, retinopathy of prematurity (ROP), retinal vein occlusion (RVO), CNV, corneal neovascularization, diseases related to corneal neovascularization, retinal neovascularization, diseases related to retinal / choroidal neovascularization, pathological myopia, von Hippel-Lindau disease, ocular histoplasmosis, familial exudative vitreoretinopathy (FEVR), Coats disease, Norrie disease, osteoporosis-pseudoglioma syndrome (OPPG), subconjunctival hemorrhage, and leukemia.

28. The drug conjugate or pharmaceutically acceptable salt thereof, or pharmaceutical composition for use according to claim 26 or 27, wherein the drug conjugate or pharmaceutically acceptable salt thereof is selected from the group consisting of baeosis, ocular neovascular diseases, neovascular glaucoma, retinitis pigmentosa (RP), hypertensive retinopathy, retinal angiomatous proliferation, macular telangiectasia, iris neovascularization, intraocular neovascularization, retinal degeneration, cystoid macular edema (CME), vasculitis, papilledema, retinitis, conjunctivitis, Leber's congenital amaurosis, uveitis, choroiditis, ocular histoplasmosis, blepharitis, dry eye, traumatic eye injury, and Sjogren's disease.

29. The drug conjugate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for use according to claims 26 to 28, wherein the ocular disorder is selected from the group consisting of AMD, DME, DR and RVO.

30. The drug conjugate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for use according to any one of claims 26 to 29, wherein the eye disorder is AMD.

31. The drug conjugate or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition for use according to any one of claims 26 to 29, wherein the AMD is wet AMD.

32. The drug conjugate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for use according to any one of claims 23 to 31, wherein the drug conjugate is administered by intraocular administration.

33. The drug conjugate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for use according to any one of claims 23 to 32, wherein the drug conjugate is administered by intraocular administration every six months.

34. The drug conjugate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition for use according to any one of claims 23 to 32, wherein the drug conjugate is administered by intraocular administration every 12 months.

35. The drug conjugate or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition for use according to any one of claims 23 to 34, wherein the drug conjugate or the pharmaceutical composition is administered by intraocular injection into the vitreous of the subject.

36. A method for preparing the drug conjugate of any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, comprising: (a) mixing solution A with solution B to form an emulsion, wherein solution A contains one or more -FG 1 and a first functionalized HA, which is optionally modified with a further functional group, and one or more -FG 2 and a second functionalized HA optionally modified with a further functional group, -FG 1 and -FG 2 are different functional group moieties, and -FG on the first functionalized HA 1 The second functionalized HA is -FG. 2 to form multiple crosslinks resulting in the formation of hydrogel HA microspheres; (b) optionally adding a pH adjuster to the emulsion of step (a); (c) collecting the resulting hydrogel HA microspheres of step (a) or (b); (d) providing the hydrogel HA microspheres or pharmaceutically acceptable salts thereof of step (c), wherein the hydrogel is free of one or more unreacted -FG 1 or -FG 2 including, steps, (e) Monoconjugate Reagent DL 1 -L 2 -FG 3 , Bisconjugate Reagent FG 3 -L 2 -L 1 -DL 1 -L 2 -FG 3 or a Tris conjugate reagent of formula (t): 【Chemistry 14】 [Wherein -D independently represents -L 1 a VEGF-neutralizing drug moiety covalently and reversibly conjugated to Each-L 1 - is independently a reversible linker moiety; Each-L 2 - is independently a spacer moiety or absent; Each-FG 3 are independently -FG 1 and -FG 2 is a functional group that reacts with preparing a (f) mixing the hydrogel HA microspheres of step (d) with the monoconjugate, bisconjugate, or trisconjugate reagent of step (e); (g) optionally, mixing the drug conjugate or pharmaceutically acceptable salt thereof of step (f) with a blocking reagent; and (h) collecting the drug conjugate or pharmaceutically acceptable salt thereof of step (f) or (g). A method comprising:

37. 37. The method of claim 36, wherein in step (a), the hydrogel HA microspheres are formed by suspension polymerization.

38. 38. The method of claim 36 or 37, wherein in step (d) the obtained microspheres have a diameter in the range of from 1 μm to 1000 μm, such as from 50 μm to 900 μm, such as from 100 μm to 700 μm, such as from 50 μm to 500 μm or such as from 200 μm to 500 μm, as determined by flow microscopy.

39. 39. The method of any one of claims 36 to 38, wherein in step (d), the obtained microspheres have a diameter in the range of 100 μm to 200 μm as determined by flow microscopy.

40. The reagent in step (e) is a monoconjugate reagent DL 1 -L 2 -FG 3 The method according to any one of claims 36 to 39, wherein

41. The blocking reagent in step (g) 【Chemistry 15】 The method of any one of claims 36 to 40, selected from the group consisting of:

42. The blocking reagent in step (g) 【Chemistry 16】 The method according to any one of claims 36 to 41, wherein

43. -FG 3 The method of any one of claims 36 to 42, wherein is -SH.

44. Monoconjugate Reagent DL 1 -L 2 -FG 3 But, equation (m1): 【Chemistry 17】 wherein -D is ranibizumab. The method according to any one of claims 36 to 43, wherein

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