Drug conjugate, pharmaceutical composition and method of treating hepatitis

Abstract

To provide a drug conjugate that is more effective than conventional drugs in the treatment of hepatitis B.SOLUTION: A drug conjugate includes a structure shown by the following formula: Z-(linker-[R]m)n. In the formula, Z is a drug compound, R is a sugar, and m and n are independently an integer from 1 to 6. The drug compound Z is a hepatitis virus targeting drug, a hepatitis B virus (HBV) drug, an inhibitor of apoptosis protein (IAP) antagonist, a multidrug resistance (MDR) inhibitor, or analogues, precursors, prodrugs, or derivatives thereof.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to drug conjugates, particularly pharmaceutical preparations containing such drug conjugates, and methods of using the drug conjugates or pharmaceutical compositions to treat Hepatitis B, Hepatitis C, viral hepatitis, or other viral infections. [Background technology]

[0002] Tenofovir and Entecavir are the first-line drugs used to treat Hepatitis B. However, these types of drugs (nucleoside / nucleotide analogues) need to be taken regularly and continuously to suppress the levels of the virus in the body. If the drugs are stopped inappropriately or if problems with the immune system occur, they usually cause a very severe viral rebound and exacerbation of severe acute hepatitis. Therefore, other treatments may be preferable.

[0003] Inhibitor of apoptosis proteins (IAPs) are a family of anti-apoptotic proteins that play a key role in evading apoptosis, and can inhibit apoptotic signaling pathways and promote survival. In addition to their use in cancer therapy, many IAP inhibitors (or IAP antagonists) are considered for the treatment of hepatitis B. For example, clinical trials of birinapant (a synthetic small molecule) in the treatment of hepatitis B are underway. However, side effects such as cranial nerve palsy (or Bell's palsy) have been observed in clinical trials when birinapant is combined with hepatitis B virus (HBV) drugs such as tenofovir and entecavir. Another IAP inhibitor, APG-1387, has been recognized for its potential in the treatment of patients with chronic hepatitis B, but further clinical trials are still ongoing. [ka]

[0004] Further research and analysis is needed to fully understand the potential of IAP inhibitors and liver targeting of carbohydrates for the treatment of hepatitis B, hepatitis C, or hepatitis-related diseases. Summary of the Invention [Problem to be solved by the invention]

[0005] In the present invention, the technical problem of the present invention is to provide a drug conjugate that is more effective than conventional drugs in treating hepatitis B. Furthermore, another problem of the present invention is to provide a pharmaceutical composition containing such a drug conjugate, and a method for treating hepatitis using the drug conjugate or the pharmaceutical composition. [Means for solving the problem]

[0006] In some embodiments of the invention, the drug conjugate has the structure shown in formula (I): Z-(Linker-[R] m ) n Formula (I) In the formula (I), Z is a drug compound, R is a sugar, and m and n are independently integers from 1 to 6.

[0007] In some embodiments, drug compound Z is a hepatitis virus targeted drug, a hepatitis B virus (HBV) drug, a hepatitis C virus (HCV) drug, an inhibitor of apoptosis protein (IAP) antagonist, a multidrug resistance (MDR) inhibitor, or an analog, precursor, prodrug, or derivative thereof.

[0008] In some embodiments, drug compound Z is selected from the group consisting of tenofovir, tenofovir diisoproxil, tenofovir alafenamide, entecavir, telbivudine, adefovir, adefovir dipivoxil, lamivudine, interferon-alpha-2A, interferon-alpha-2B, birinapant, monomeric birinapant, zebinapant, LCL161, GDC-0152, GDC-0917, CUDC-427, APG-1387, DeBio-1143, sergantolimod, BI- 82, glecaprevir, pibrentasvir, ombitasvir, paritaprevir, ritonavir, sovosbuvir, velapatasvir, ledipasvir, voxilaprevir, daclatasvir, asunaprevir, telaprevir, elbasvir, grazoprevir, ribavarin, telaprevir, EDP-239, alisporivir, filibirubir, melicitabine, danoprevir, velpatasvir, zosuquidar, or an analogue, precursor, prodrug, or derivative thereof.

[0009] In some embodiments, the sugar R is selected from the group consisting of a monosaccharide, a disaccharide, a trisaccharide, a tetrasaccharide, an oligosaccharide, a polysaccharide, or derivatives thereof.

[0010] In some embodiments, the linker is selected from the group consisting of substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, aryl alkyl, aryl alkenyl, aryl alkynyl, heteroaryl alkyl, heteroaryl alkenyl, heteroaryl alkynyl, heterocyclyl alkyl, heterocyclyl alkenyl, heterocyclyl alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylaryl alkyl, alkylaryl alkenyl, alkylaryl alkynyl, alkenylaryl alkyl, alkenylaryl alkynyl, alkynylaryl alkyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkylheteroaryl alkyl, alkylheteroaryl alkenyl, alkylheteroaryl a and wherein one or more methylenes are selected from the group consisting of O, S, S(O), SO, N(R), S ... 8 ), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, wherein R 8 is hydrogen, acyl, aliphatic or substituted aliphatic.

[0011] In some embodiments, the linker is any one selected from the group consisting of formula (LX1) to formula (LX30). [ka] JPEG2025009924000004.jpg241168JPEG2025009924000005.jpg252167JPEG2025009924000006.jpg255163JPEG2025009924000007.jpg255156JPEG2025009924000008.jpg212169JPEG2025009924000009.jpg255169JPEG2025009924000010.jpg157170JPEG2025009924000011.jpg151167In the formulas (LX1) to (LX30), Z is the position where the linker is bonded to the drug, and R is the position where the linker is bonded to the sugar.

[0012] In some embodiments, the sugar R is selected from the group consisting of a monosaccharide, a disaccharide, and a trisaccharide, which are independently unsubstituted or substituted with an acetyl or N-acetyl group, and wherein the drug Z is selected from the group consisting of entecavir, adefovir, telbivudine, lamivudine, tenofovir, GDC-0152, DeBio-1143, LCL161, birinapant, or analogs, precursors, prodrugs, or derivatives thereof.

[0013] In some embodiments, in the drug conjugate, the drug compound Z is attached to the linker to form a carbamate bond (>NC(=O)-O-), an amide bond (RC(=O)-N<), a carbonate bond (RO-C(=O)-OR'), or an ester bond (RC(=O)-OR').

[0014] In some embodiments, the linker comprises at least a triazole.

[0015] In some embodiments, the structure shown in formula (I) is further represented by formula (II): Z-(Linker-[R] m )2 formula (II) In the formula (II), Z is a drug compound, R is a sugar, and m is independently an integer of 1 to 3.

[0016] In some embodiments, the structure shown in formula (I) is further represented by any one of formulas (IA1)-(IA4). [ka]

[0017] In some embodiments, the structure shown in formula (I) is further represented by any one of formulas (IB1)-(IB11). [ka] JPEG2025009924000014.jpg204170JPEG2025009924000015.jpg241169JPEG2025009924000016.jpg227167JPEG2025009924000017.jpg215164

[0018] In some embodiments, the structure shown in formula (I) is further represented by any one of formulas (IC1)-(IC11). [ka] JPEG2025009924000019.jpg251169JPEG2025009924000020.jpg172170

[0019] In some embodiments, the structure shown in formula (I) is further represented by any one of formulas (ID1)-(ID5). [ka] JPEG2025009924000022.jpg60170

[0020] In some embodiments, the structure shown in formula (I) is further represented by formula (IE1). [ka]

[0021] In some embodiments, the structure shown in formula (I) is further represented by formula (IF1) or formula (IF2). [ka]

[0022] In some embodiments, the structure shown in formula (I) is further represented by formula (IG1): [ka]

[0023] In some embodiments, the structure shown in formula (I) is further represented by any one of formulas (IH1) through (IH3). [ka]

[0024] In some embodiments, the structure shown in formula (I) is further represented by formula (IJ1) or formula (IJ2). [ka]

[0025] In some embodiments of the present invention, the pharmaceutical composition comprises an active ingredient comprising a drug conjugate having the structure shown in formula (I). Z-(Linker-[R] m ) n Formula (I) In the formula (I), Z is a drug compound, R is a sugar, and m and n are independently integers from 1 to 6; At least one or more pharma- ceutically acceptable excipients.

[0026] In some embodiments, the drug conjugate comprises a first drug conjugate represented by Formula (I), where drug compound Z is a first drug compound X selected from the group consisting of tenofovir, tenofovir diisoproxil, tenofovir alafenamide, entecavir, telbivudine, adefovir, adefovir dipivoxil, lamivudine, interferon-alpha-2A, interferon-alpha-2B, sergantolimod, BI-82, and zosquidar, or analogs, precursors, prodrugs, derivatives thereof, or any HBV drug.

[0027] In some embodiments, the first drug conjugate represented by Formula (I) is further represented by any one of Formulas (IA1)-(IA4), any one of Formulas (IC1)-(IC11), or Formula (IF1), or Formula (IF2), or Formula (IG1). [ka] JPEG2025009924000029.jpg249169JPEG2025009924000030.jpg195169JPEG2025009924000031.jpg219169JPEG2025009924000032.jpg144170

[0028] In some embodiments, the drug conjugate further comprises a second drug conjugate represented by formula (I), wherein drug compound Z is a second drug compound Y selected from the group consisting of birinapant, monomeric birinapant, zebinapant, LCL161, GDC-0152, GDC-0917, CUDC-427, APG-1387, DeBio-1143, or analogs, precursors, prodrugs, derivatives thereof, or any IAP inhibitor.

[0029] In some embodiments, the second drug conjugate represented by Formula (I) is further represented by any one of Formulas (IB1)-(IB11), or any one of Formulas (ID1)-(ID5), any one of Formulas (IH1)-(IH3), or Formula (IJ1) or Formula (IJ2). [ka] JPEG2025009924000034.jpg195167JPEG2025009924000035.jpg244167JPEG2025009924000036.jpg228169JPEG20250099240 00037.jpg214166JPEG2025009924000038.jpg228169JPEG2025009924000039.jpg237170JPEG2025009924000040.jpg129169

[0030] In some embodiments, the first drug conjugate is a compound represented by Formula (IA1) and the second drug conjugate is a compound represented by Formula (IB1). [ka]

[0031] In some embodiments, the first drug conjugate and the second drug conjugate are each administered at a dose of 0.1 mg / kg to 100 mg / kg.

[0032] In some embodiments, the sugar R is selected from the group consisting of a monosaccharide, a disaccharide, a trisaccharide, a tetrasaccharide, an oligosaccharide, a polysaccharide, or derivatives thereof.

[0033] In some embodiments, the linker is selected from the group consisting of substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, aryl alkyl, aryl alkenyl, aryl alkynyl, heteroaryl alkyl, heteroaryl alkenyl, heteroaryl alkynyl, heterocyclyl alkyl, heterocyclyl alkenyl, heterocyclyl alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylaryl alkyl, alkylaryl alkenyl, alkylaryl alkynyl, alkenylaryl alkyl, alkenylaryl alkynyl, alkynylaryl alkyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkylheteroaryl alkyl, alkylheteroaryl alkenyl, alkylheteroaryl a and wherein one or more methylenes are selected from the group consisting of O, S, S(O), SO, N(R), S ... 8 ), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, wherein R 8 is hydrogen, acyl, aliphatic or substituted aliphatic.

[0034] In some embodiments, the pharmaceutical composition is prepared into a tablet, capsule, granule, powder, solution, syrup, spray, injection or inhalant.

[0035] In some embodiments, the at least one or more pharma- ceutically acceptable excipients are selected from the group consisting of fillers, bulking agents, binders, mixing agents, surfactants, emulsifiers, dispersing agents, antifoaming agents, lubricants, non-adherents, mixing agents, coating materials, glidants, anti-adherents, diluents, dyes, pigments, dispersing agents, wetting agents, and combinations thereof.

[0036] According to some embodiments, a method for treating hepatitis (e.g., hepatitis B, hepatitis C, viral hepatitis, etc.) is described, comprising administering to a patient with hepatitis a therapeutically effective amount of a drug conjugate.

[0037] According to some embodiments, a method for treating hepatitis is described that comprises administering a pharmaceutical composition to a patient suffering from hepatitis. Effect of the Invention

[0038] According to the above embodiment, the drug conjugate or pharmaceutical composition of the present invention is more effective than conventional drugs in treating hepatitis B. For example, the drug conjugate or pharmaceutical composition can further reduce hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) levels and HBV DNA levels while improving hepatitis B surface antibody (anti-HBsAg) levels compared to conventional hepatitis B treatment methods. From preliminary evaluation, the inventors have also found that modifying HCV drugs to form drug conjugates with similar linkers and sugars may also be promising for providing a more effective treatment for hepatitis C.

[0039] In order to make the foregoing more easily understandable, several embodiments accompanied with drawings are described in detail below. [Brief description of the drawings]

[0040] The accompanying drawings are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification. The drawings illustrate exemplary embodiments of the invention and, together with the description, serve to explain the principles of the invention. [Figure 1]1 is a scheme showing the synthesis of a drug conjugate of formula (IA1). [Diagram 2] 1 is a scheme showing the synthesis of a drug conjugate of formula (IB1). [Diagram 3] 1 is a scheme showing the synthesis of a drug conjugate of formula (IC1). [Figure 4] 1 is a scheme showing the synthesis of a drug conjugate of formula (ID1). [Diagram 5] 1 is a scheme showing the synthesis of a drug conjugate of formula (IE1). [Figure 6] 1 is a scheme showing the synthesis of a drug conjugate of formula (IF1). [Figure 7] 1 is a scheme showing the synthesis of a drug conjugate of formula (IG1). [Figure 8] 1 is a scheme showing the synthesis of a drug conjugate of formula (IH1). [Figure 9] 1 is a scheme showing the synthesis of a drug conjugate of formula (IJ1). [Figure 10] FIG. 10 is a graph showing HBsAg levels from in vitro cell line experiments for different entecavir-based Hepatitis B treatment groups. [Figure 11] 1 is a graph showing HBsAg levels from in vitro cell line experiments for different birinapant-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 12] FIG. 1 is a graph showing HBsAg levels from in vitro cell line experiments for different tenofovir-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 13] 1 is a graph showing HBsAg levels from in vitro cell line experiments for different LCL161-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 14] 1 is a graph showing HBsAg levels from in vitro cell line experiments for different telbivudine-based or adefovir-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 15]1 is a graph showing HBsAg and HBeAg levels from in vitro cell line experiments for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 16A] 1 is a graph showing HBsAg and HBeAg levels from in vitro cell line experiments for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 16B] 1 is a graph showing Hepatitis B virus (HBV) DNA levels from in vitro cell line experiments for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 17A] 1 is a graph showing HBsAg levels from animal studies for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 17B] 1 is a graph showing HBsAg levels from animal studies for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 18A] 1 is a graph showing HBeAg levels from an animal study for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 18B] 1 is a graph showing HBeAg levels from an animal study for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 19A] 1 is a graph showing HBV DNA levels from animal studies for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 19B] 1 is a graph showing HBV DNA levels from animal studies for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 20] 1 shows the observation of hair loss in C3H mice after administration of different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 21] 1 is a graph showing anti-HBsAg levels from an animal study for different Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 22A]1 is a graph showing HBsAg levels from animal studies for entecavir-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 22B] 1 is a graph showing HBsAg levels from animal studies for entecavir-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 23A] 1 is a graph showing HBeAg levels from an animal study for entecavir-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 23B] 1 is a graph showing HBeAg levels from an animal study for entecavir-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 24A] 1 is a graph showing HBV DNA levels from animal studies for entecavir-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 24B] 1 is a graph showing HBV DNA levels from animal studies for entecavir-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 25A] 1 is a graph showing HBsAg levels from animal studies for birinapant-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 25B] 1 is a graph showing HBsAg levels from animal studies for birinapant-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 26A] 1 is a graph showing HBeAg levels from animal studies for birinapant-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 26B] 1 is a graph showing HBeAg levels from animal studies for birinapant-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 27A] 1 is a graph showing HBV DNA levels from animal studies for birinapant-based Hepatitis B treatment groups according to some embodiments of the present invention. [Figure 27B]1 is a graph showing HBV DNA levels from animal studies for birinapant-based Hepatitis B treatment groups according to some embodiments of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0041] The present invention describes a drug conjugate having the structure shown in formula (I). Z-(Linker-[R] m ) n Formula (I) In the formula (I), Z is a drug compound, R is a sugar, and m and n are independently integers from 1 to 6.

[0042] In exemplary embodiments, the drug compound Z is a hepatitis virus targeted drug, a hepatitis B virus (HBV) drug, a hepatitis C virus (HCV) drug, an inhibitor of apoptosis protein (IAP) antagonist, a multidrug resistance (MDR) inhibitor, or an analog, precursor, prodrug, or derivative thereof. In exemplary embodiments, the drug conjugate is used to treat hepatitis B, so the drug compound Z used is selected from an HBV drug or an IAP antagonist. In some other embodiments, the drug conjugate is used to treat hepatitis C. In some other embodiments, the drug conjugate is used to treat human immunodeficiency virus (HIV). However, the present invention is not so limited. In some alternative embodiments, when the drug conjugate is used to treat other diseases, other types of drugs may be used to form the drug conjugate shown in formula (I).

[0043] In some embodiments, when drug compound Z is an HBV drug or an IAP antagonist, drug compound Z is selected from the group consisting of tenofovir, tenofovir diisoproxil, tenofovir alafenamide, entecavir, telbivudine, adefovir, adefovir dipivoxil, lamivudine, interferon-alpha-2A, interferon-alpha-2B, birinapant, monomeric birinapant, zebinapant, LCL161, GDC-0152, GDC-0917, CUDC-427, APG-1387, sergantolimod, BI-82, and zosquidar, or analogs, precursors, prodrugs, or derivatives thereof. For example, tenofovir, tenofovir diisoproxil, tenofovir alafenamide, entecavir, telbivudine, adefovir, adefovir dipivoxil, lamivudine, interferon-α-2A, interferon-α-2B, sergantolimod, BI-82, and zosuquidar are HBV drugs, and birinapant, monomeric birinapant, zebinapant, LCL161, GDC-0152, GDC-0917, CUDC-427, and APG-1387 are IAP antagonists. However, the present invention is not limited thereto, and other HBV drugs, IAP antagonists, or analogs, precursors, and derivatives thereof known in the art may be exemplified.

[0044] In some embodiments, when drug compound Z is an HCV drug, drug compound Z is selected from the group consisting of glecaprevir, pibrentasvir, ombitasvir, paritaprevir, ritonavir, sovosbuvir, velapatasvir, ledipasvir, voxilaprevir, daclatasvir, asunaprevir, telaprevir, elbasvir, grazoprevir, ribavarin, telaprevir, EDP-239, alisporivir, filibirubir, melicitabine, danoprevir, and velpatasvir, or analogs, precursors, prodrugs, or derivatives thereof.

[0045] HBV drugs (or their analogues, precursors, or derivatives): [ka]

[0046] IAP antagonists (or their analogs, precursors, derivatives): [ka]

[0047] In some embodiments, the sugar R is selected from the group consisting of a monosaccharide, a disaccharide, a trisaccharide, a tetrasaccharide, an oligosaccharide, a polysaccharide, or derivatives thereof. The term "monosaccharide" may include allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, D-fucitol, L-fucitol, fucosamine, fucose, fuculose, galactosamine, D-galactosaminitol, N-acetylgalactosamine, galactose, glucosamine, N-acetylglucosamine, glucosaminitol, glucose, glucose-6-phosphate, guloseglyceraldehyde, L-glycero-D-mannose-heptose, glycerol, glycerone, gulose, idose, lyxose, mannosamine, mannose, mannose-6-phosphate, psicose, quinovose, quinovosamine, rhamnitol, rhamnosamine, rhamnose, ribose, ribulose, sedoheptulose, sorbose, tagatose, talose, tartaric acid, threose, xylose, and xylulose. Monosaccharides may be in the D- or L-configuration. Monosaccharides may further be deoxysugars (where the alcoholic hydroxy group is replaced with hydrogen), aminosugars (where the alcoholic hydroxy group is replaced with an amino group), thiosugars (where the alcoholic hydroxy group is replaced with a thiol, or the C=O is replaced with a C=S, or the ring oxygen is replaced with sulfur), selenosugars, tellurosugars, azasugars (where a ring carbon is replaced with nitrogen), iminosugars (where a ring oxygen is replaced with nitrogen), phosphanosugars (where a ring oxygen is replaced with phosphorus), phosphasugars (where a ring carbon is replaced with phosphorus), C-substituted monosaccharides (where a hydrogen on a non-terminal carbon atom is replaced with carbon), unsaturated monosaccharides, alditols (where a carbonyl group is replaced with a CHOH group), aldonic acids (where an aldehyde group is replaced with a carboxy group), ketoaldonic acids, uronic acids, aldaric acids, and the like. Amino sugars include amino monosaccharides, preferably galactosamine, glucosamine, mannosamine, fucosamine, quinovosamine, neuraminic acid, muramic acid, lactosediamine, acosamine, bacillosamine, daunosamine, desosamine, forosamine, galosamine, kanosamine, kansosamine, mycaminose, mycosamine, perosamine, pneumosamine, purpurosamine, rhodosamine.In some embodiments, the monosaccharides etc. may be further substituted.

[0048] In some embodiments, in Formula (I), the linker is selected from the group consisting of substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, aryl alkyl, aryl alkenyl, aryl alkynyl, heteroaryl alkyl, heteroaryl alkenyl, heteroaryl alkynyl, heterocyclyl alkyl, heterocyclyl alkenyl, heterocyclyl alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylaryl alkyl, alkylaryl alkenyl, alkylaryl alkynyl, alkenylaryl alkyl, alkenylaryl alkynyl, alkynylaryl alkyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkylheteroaryl alkyl, alkylheteroaryl alkenyl ... wherein one or more methylenes are selected from the group consisting of O, S, S(O), SO, N(R), S ... 8 ), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, wherein R 8 is hydrogen, acyl, aliphatic or substituted aliphatic.

[0049] In some embodiments, one, two or more linkers and one, two or more sugars R may be attached to the drug compound Z (e.g., n is 1-6). Furthermore, one, two or more sugars R may be attached to a linker (e.g., m is 1-6). That is, the drug compound Z may be monovalent, divalent, trivalent or polyvalent. When two or more sugars R and two or more linkers are used in the drug conjugate, the sugars R and the linkers may be independently the same or different. Although an exemplary embodiment shows one linker attached to one sugar R, the invention is not limited thereto. In some alternative embodiments, two or more sugars R or two or more drug compounds Z may be attached to one linker. That is, the linker may have a branched structure. In certain embodiments, in the drug conjugate, the drug compound Z is attached to the linker to form a carbamate bond (>NC(=O)-O-). Here, the carbamate bond may include a portion of the drug compound Z and a portion of the linker. In certain embodiments, the linker includes at least a triazole. In some other embodiments, in the drug conjugate, the drug compound Z is bonded to the linker to form a carbamate bond (>NC(=O)-O-), an amide bond (RC(=O)-N<), a carbonate bond (RO-C(=O)-OR'), or an ester bond (RC(=O)-OR').

[0050] In some embodiments, the linker of formula (I) is L 1 (L 2 ) p group, where L group is attached to the drug compound Z and L 2 The base is L 1 The L group is attached to the sugar R, where p is an integer from 1 to 4. 1 The group is L 1The group may comprise a carbamate, amide, ester, amidine, carboximidate, sulfonate, sulfate, sulfonamide, thioether, ether, imine, imide, thioamide, carbonate, thiocarbamide, carbamide, triazole, amine, oxime, thiocarbonate, thioester, or thiocarbamate group attached to the drug compound Z. 2 The group is a single bond, an alkyl group having 1 to 50 carbon atoms, an alkoxy group having 1 to 50 carbon atoms, an alkenyl group having 2 to 50 carbon atoms, or an alkenyloxy group having 2 to 50 carbon atoms, and in the above groups, the alkyl group, the alkoxy group, the alkenyl group, and the alkenyloxy group may be linear or branched, at least one -CH2- may be substituted with -CO-, -COO-, -OCO-, -S-, -O-, or -NH-, and at least one -CH2CH2- may be substituted with -CH=CH- or -C≡C-, and the above-mentioned -CH2-, -CH=, or At least one hydrogen in -NH- may be substituted with halogen, trifluoromethyl, alkyl having 1 to 20 carbons, alkoxy having 1 to 20 carbons, or alkanoyl having 1 to 20 carbons, and at least one -O- described above may be substituted with a phosphate, phosphonate, phosphonate, phosphinate, phosphine oxide, phosphoamide, phosphoramidate, phosphite, phosphonite, phosphine, aminophosphine, phosphoramidite, phosphonamidite, phosphonamide, phosphinamide, or phosphorodiamidite group.

[0051] In certain embodiments, L 1 The group is a carbamate, amide, carbonate, or ester; L 2The group is an alkyl group having 1 to 20 carbons, an alkoxy group having 1 to 20 carbons, an alkenyl group having 2 to 20 carbons, or an alkenyloxy group having 2 to 20 carbons. In the above groups, the alkyl group, the alkoxy group, the alkenyl group, and the alkenyloxy group may be linear or branched, and at least one -CH2- group may be substituted with -CO-, -COO-, -OCO-, -S-, -O-, or -NH-, and at least one -CH2CH2- group may be substituted with -CH=CH-.

[0052] In some embodiments, the linker has the structure: [ka] JPEG2025009924000045.jpg233170JPEG2025009924000046.jpg252168JPEG2025009924000047.jpg252161JPEG2025009924000048.jpg242168 JPEG2025009924000049.jpg192170JPEG2025009924000050.jpg243169JPEG2025009924000051.jpg156170JPEG2025009924000052.jpg151168

[0053] In the above linkers (LX1) to (LX30), Z is the position where the linker is attached to the drug, and R is the position where the linker is attached to the sugar. In some embodiments, the sugar R may be attached to the linker via any free hydroxyl group / free leaving group on the sugar R. That is, the position where the linker is attached to the sugar is not particularly limited, so long as the linker is covalently attached to the sugar R. Similarly, the linker may be attached to the drug compound Z via any free leaving group on the drug compound Z. In some other embodiments, the linker may be omitted. That is, the drug compound Z may be directly attached to the sugar R.

[0054] In some embodiments, the structure shown in formula (I) is further represented by formula (II): Z-(Linker-[R] m )2 formula (II) In the formula (II), Z is a drug compound, R is a sugar, and m is independently an integer of 1 to 3.

[0055] That is, in formula (II), there are two sugar chains attached to drug Z via a linker, and the sugars of the two sugar chains are selected from monosaccharides, disaccharides, or trisaccharides.

[0056] In some embodiments, the structure shown in formula (I) is an entecavir-based compound further represented by any one of formulas (IA1)-(IA3). [ka]

[0057] In some other embodiments, the structure of formula (I) is an entecavir-based compound further represented by any one of formulas (IA4)-(IA13). [ka] JPEG2025009924000055.jpg197169JPEG2025009924000056.jpg84167

[0058] In some embodiments, the structure shown in formula (I) is a birinapant compound further represented by any one of formulas (IB1)-(IB5). [ka] JPEG2025009924000058.jpg172137JPEG2025009924000059.jpg200165JPEG2025009924000060.jpg110169

[0059] In some other embodiments, the structure of formula (I) is a birinapant compound further represented by any one of formulas (IB6)-(IB19). [ka] JPEG2025009924000062.jpg219169JPEG2025009924000063.jpg247168JPEG2025009924000064.jpg239169JPEG2025009924000065.jpg204168

[0060] In some embodiments, the structure of formula (I) is a tenofovir compound further represented by any one of formulas (IC1)-(IC11). [ka] JPEG2025009924000067.jpg248170JPEG2025009924000068.jpg173169

[0061] In some embodiments, the structure shown in formula (I) is an LCL161-based compound further represented by any one of formulas (ID1)-(ID4). [ka]

[0062] In some other embodiments, the structure of formula (I) is an LCL161-based compound further represented by any one of formulas (ID5) and (ID6). [ka]

[0063] In some embodiments, the structure shown in formula (I) is a telbivudine compound further represented by formula (IE1). [ka]

[0064] In some embodiments, the structure of formula (I) is an adefovir compound further represented by formula (IF1): [ka]

[0065] In some other embodiments, the structure of formula (I) is an adefovir compound further represented by formula (IF2). [ka]

[0066] In some embodiments, the structure shown in formula (I) is a lamivudine-based compound further represented by any one of formulas (IG1) and (IG2). [ka]

[0067] In some embodiments, the structure shown in formula (I) is a GDC-0152-based compound represented by any one of formulas (IH1)-(IH4). [ka]

[0068] In some embodiments, the structure shown in formula (I) is a DeBio-1143-based compound represented by any one of formulas (IJ1) and (IJ2). [ka]

[0069] In some other embodiments, the pharmaceutical composition comprises an active ingredient (active pharmaceutical ingredient) and at least one or more pharma- ceutically acceptable excipients. For example, the active ingredient comprises a drug conjugate having the structure shown in formula (I) above. In some embodiments, the drug conjugate comprises a first drug conjugate represented by formula (I). In formula (I), drug compound Z is a first drug compound X selected from the group consisting of tenofovir, tenofovir diisoproxil, tenofovir alafenamide, entecavir, telbivudine, adefovir, adefovir dipivoxil, lamivudine, interferon-α-2A, interferon-α-2B, sergantolimod, BI-82, and zosquidar, or analogs, precursors, prodrugs, and derivatives thereof. For example, the first drug conjugate represented by formula (I) may further be represented by any of formulas (IA1)-(IA3), any of formulas (IC1)-(IC11), formula (IE1), or formula (IF1) above. In some embodiments, the drug conjugate comprises a second drug conjugate represented by formula (I). In formula (I), drug compound Z is a second drug compound Y selected from the group consisting of birinapant, monomeric birinapant, zebinapant, LCL161, GDC-0152, GDC-0917, CUDC-427, APG-1387, or analogs, precursors, prodrugs, and derivatives thereof. For example, the second drug conjugate represented by formula (I) is further represented by any of formulas (IB1) to (IB5) or any of formulas (ID1) to (ID4) above.

[0070] In some embodiments, a pharmaceutical composition can effectively improve the treatment of Hepatitis B by combining a drug conjugate comprising a first drug compound X (HBV drug) and a second drug compound Y (IAP antagonist). In an exemplary embodiment, the first drug conjugate comprising the first drug compound X is a compound represented by formula (IA1), and the second drug conjugate comprising the second drug compound Y is a compound represented by formula (IB1). [ka]

[0071] In an exemplary embodiment, the first drug conjugate and the second drug conjugate are each administered at a dose of 0.1 mg / kg to 100 mg / kg. In some embodiments, the pharmaceutical composition is prepared into a tablet, capsule, granule, powder, solution, syrup, spray, injection, or inhalant. In some embodiments, the pharma- ceutical composition includes a pharma- ceutical composition comprising a pharma- ceutical composition selected from the group consisting of a filler, a bulking agent, a binder, a mixing agent, a surfactant, an emulsifier, a dispersing agent, an antifoaming agent, a lubricant, a non-adhesive agent, a mixing agent, a coating material, a glidant, an anti-adherent, a diluent, a dye, a pigment, a dispersing agent, a wetting agent, and combinations thereof. For example, the excipients may be used alone or in combination.

[0072] In some other embodiments, a method for treating hepatitis (such as hepatitis B, hepatitis C, viral hepatitis, etc.) is described. For example, the method includes administering a therapeutically effective amount of a drug conjugate to a patient with hepatitis (such as hepatitis B, hepatitis C, viral hepatitis, etc.) or administering a pharmaceutical composition to a patient with hepatitis (such as hepatitis B, hepatitis C, viral hepatitis, etc.). By administering the drug conjugate or pharmaceutical composition of the present invention, hepatitis B surface antibody (anti-HBsAg) levels can be further improved while further reducing hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) levels, and / or hepatitis B virus (HBV) DNA levels compared to conventional hepatitis B treatment methods. Similarly, modifying HCV drugs to form drug conjugates with similar linkers and sugars may also hold promise for providing more effective treatment of hepatitis C.

[0073] Working Example To demonstrate that the drug conjugate or pharmaceutical composition of the present invention is superior to conventional methods in treating hepatitis, such as hepatitis B, the following experiment was performed.

[0074] Synthesis Example The drug conjugates used in the examples were synthesized according to the following examples.

[0075] Synthesis Example A1: Synthesis of Drug Conjugate of Formula (IA1)

[0076] The drug conjugate of formula (IA1) was synthesized according to the scheme shown in Figure 1. Specifically, a solution of (3R,4R,5S,6R)-6-(hydroxymethyl)-5-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2,3,4-triol (20.00 g, 55.51 mmol) and sodium acetate (9.11 g, 111.02 mmol) in AcO (60 mL, 634.71 mmol) was refluxed at 90 °C for 2 h. The mixture was diluted with NaHCO 3( (aqueous) and extracted with EtOAc. The combined organics were dried over MgSO4 and concentrated. The crude product compound 3 was used in the next step without purification.

[0077] To a solution of compound 3 ((3R,4S,5R,6R)-6-(acetoxymethyl)-5-(((2S,3R,4S,5S,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2,3,4-triyl triacetate) (8.89 g, 13.09 mmol), compound 1 (6-azidohexan-1-ol) (7.91 g, 55.23 mmol) and 4 Å MS (1.00 g) in DCM (70 mL) was added BF3OEt2 (6.6 mL, 52.38 mmol) at 0° C. and stirred for 15 minutes, then allowed to warm to room temperature over 12 hours. The mixture was quenched with NaHCO3 (aq) and extracted with DCM. The combined organics were dried over MgSO4 and concentrated. The residue was purified by silica gel column chromatography using EtOAc:hexane (1:2) as eluent to give compound 4 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-azidohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (3.34 g, 4.46 mmol, 34% yield).

[0078] To a solution of compound 4 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-azidohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (500 mg, 0.66 mmol) in THF (5 mL) was added a solution of PMe3 in THF (1.32 mL, 1.32 mmol) at 0 °C. After warming to room temperature, the reaction mixture was stirred for 12 h. The mixture was concentrated under reduced pressure and the solvent was removed by evaporation. The crude product compound 5 was used in the next step without purification.

[0079] To a solution of compound 54 ((1S,2R,4S)-4-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)-2-(hydroxymethyl)-3-methylenecyclopentyl acetate) (195 mg, 0.61 mmol) in THF (3 mL) and DMF (1 mL) was added a solution of NMM (338 μL, 2.42 mmol) and 4-nitrophenyl chloroformate (244 mg, 1.21 mmol) in THF (2 mL) at 0 °C. The reaction mixture was then stirred at room temperature for 12 h. To the reaction mixture was added a solution of compound 5 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-aminohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (365 mg, 0.50 mmol) in THF (3 mL), and the mixture was further stirred at room temperature for 12 hours. The mixture was concentrated and purified by silica gel column chromatography using EtOAc:hexane (1:1) as eluent to give the product compound 89 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-6-((6-(((((1R,3S,5S)-5-acetoxy-3-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)-2-methylenecyclopentyl)methoxy)carbonyl)amino)hexyl)oxy)-2-(acetoxymethyl)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (193 mg, 0.17 mmol, 34% yield).

[0080] To a solution of compound 89 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-6-((6-(((((1R,3S,5S)-5-acetoxy-3-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)-2-methylenecyclopentyl)methoxy)carbonyl)amino)hexyl)oxy)-2-(acetoxymethyl)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (193 mg, 0.17 mmol) in MeOH (2 mL) was added NaOMe (15 mg, 0.28 mmol) at 0 °C. After warming to room temperature, the reaction mixture was stirred for 12 h. The mixture was neutralized with Amberlite IR-120(H) to pH 7. The solution was filtered and the solvent was removed. The resulting material was purified by reverse phase silica gel column chromatography to give the product compound 90 (((1R,3S,5S)-3-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)-5-hydroxy-2-methylenecyclopentyl)methyl(6-(((2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexyl)carbamate) (40 mg, 0.05 mmol, 30% yield). Compound 90 corresponds to the structure of formula (IA1).

[0081] NMR analysis of compound 90 is as follows: 1H-NMR (400 MHz, CD3OD): δ7.74(s, 1H), 5.54-5.48(m, 1H), 5.29-5.27(m, 1H), 4.57(s, 1H), 4.37-4.26(m, 6H), 3.92-3.67(m, 8H), 3.60-3.46(m, 8H), 3.42-3.37(m, 1H), 3.26-3.21(m, 1H), 3.14-3.09(m, 2H), 2.88-2.83(m, 1H), 2.54-2.45(m, 1H), 2.26-2.19(m, 1H), 1.65-1.34(m, 10H). NMR results confirmed that the structure of formula (IA1) was obtained. ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (IA1).

[0082] Synthesis Examples A2 to A4: Synthesis of Drug Conjugates of Formulae (IA2) to (IA4)

[0083] In Synthesis Examples A2 to A4, the drug conjugates of formula (IA2) to (IA4) were synthesized using the same organic synthesis techniques as those described in Synthesis Example A1, and the details of the synthesis are omitted here. NMR and ESI-MS analyses were performed to prove that the drug conjugates of formula (IA2) to (IA4) were successfully obtained.

[0084] The NMR analysis of the resulting drug complex of formula (IA2) is as follows: 1 H-NMR (400 MHz, DO): δ 7.96-7.80 (br, 2H), 5.53-4.97 (br, 7H), 4.54-3.99 (m, 10H), 3.97-3.37 (m, 28H), 3.31-2.97 (m, 10H), 2.87-2.77 (m, 3H), 2.39-1.76 (m, 10H), 1.59-1.1 (m, 20H). NMR confirmed that the structure of formula (IA2) was obtained. ESI-MS also confirmed the presence of peaks corresponding to the molecular weight of the structure of formula (IA2).

[0085] The NMR analysis of the resulting drug complex of formula (IA3) is as follows: 1H-NMR (400 MHz, CD3OD): δ8.07(s, 1H), 7.74(s, 1H), 5.52-5.48(m, 1H), 5.28-5.27(m, 2H), 5.25-5.24(m, 1H), 4.79-4.78(m, 1H), 4.49-4.28(m, 8H), 3.92-3.68(m, 7H), 3.62-3.48(m, 7H), 3.43-3.39(m, 1H), 3.27-3.23(m, 1H), 2.91-2.88(m, 1H), 2.47-2.40(m, 1H), 2.27-2.21(m, NMR confirmed that the structure of formula (IA3) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IA3).

[0086] The NMR analysis of the resulting drug complex of formula (IA4) is as follows: 1 H-NMR (400 MHz, CD3OD): δ7.75(s, 1H), 7.72(s, 1H), 5.52-5.48(m, 1H), 5.27-5.26(m, 1H), 4.38-4.34(m, 7H), 4.28-4.26(m ,1H), 3.91-3.68(m, 7H), 3.60-3.47(m, 7H), 3.41-3.37(m, 1H), 3.26-3.21(m, 1H), 2.90-2.85(m, 1H), 2.77-2.73(m, 2H), 2.53-2.46(m, 1H), 2.45-2.41(m, NMR results confirmed that the structure of formula (IA4) was obtained. ESI-MS also confirmed peaks corresponding to the molecular weight of the structure of formula (IA4).

[0087] Synthesis Example B1: Synthesis of a drug conjugate of formula (IB1)

[0088] The drug conjugate of formula (IB1) was synthesized according to the scheme shown in Figure 2. Specifically, ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-(benzyloxy)-6-oxohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (600 mg, 0.71 mmol) in MeOH (5 mL) was added with 10% palladium on carbon (60 mg, 0.1 w%) and stirred under H2 for 3 h. The mixture was filtered through Celite to remove palladium on carbon, concentrated under reduced pressure and evaporated to obtain the crude product compound (compound 24). The crude product compound 24 was used in the next step without purification.

[0089] To a solution of compound 80 (dibenzyl 5,5′-((6,6′-difluoro-1H,1′H-[2,2′-biindole]-3,3′-diyl)bis(methylene))(3S,3′S,5R,5′R)-bis(3-acetoxypyrrolidine-1-carboxylate)) (300 mg, 0.34 mmol) in THF (30 mL) was added palladium on carbon 10 wt.% (30 mg). The reaction mixture was stirred under hydrogen at room temperature for 2 hours. After the reaction was completed, the resulting material was filtered and concentrated. The desired product was obtained as a yellow solid ((3S,3′S,5R,5′R)-((6,6′-difluoro-1H,1′H-[2,2′-biindole]-3,3′-diyl)bis(methylene))bis(pyrrolidine-5,3-diyl)diacetate) (equal yield). The resulting compound is designated as Compound 81.

[0090] To a solution of compound 81 (132 mg, 0.24 mmol) and compound 24 (6-(((2R,3R,4S,5R,6R)-3,4-diacetoxy-6-(acetoxymethyl)-5-(((2S,3R,4S,5S,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexanoic acid) (533 mg, 0.71 mmol) in DCM (10 mL) was added HATU (280 mg, 0.74 mmol) and DIPEA (800 μL, 5.87 mmol) and stirred at room temperature for 12 h. The mixture was purified by silica gel column chromatography to give a compound designated as compound 133 (450 mg, 0.22 mmol, 93% yield).

[0091] To a solution of compound 133 (450 mg, 0.21 mmol) in MeOH (6 mL) and DCM (6 mL) was added NaOMe (32 mg, 0.60 mmol) at 0° C. After warming to room temperature, the reaction mixture was stirred for 1 h. The mixture was quenched with Amberlite IR-120 (H) and filtered off. The resulting material was purified by reverse phase silica gel column chromatography to give the product compound 134 (1,1'-((3S,3'S,5R,5'R)-((6,6'-difluoro-1H,1'H-[2,2'-biindole]-3,3'-diyl)bis(methylene))bis(3-hydroxypyrrolidine-5,1-diyl))bis(6-(((2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexan-1-one)) (200 mg, 0.15 mmol, 67% yield). Compound 134 corresponds to the structure of formula (IB1).

[0092] The NMR analysis of compound 134 (formula (IB1)) is as follows: 1H-NMR (400 MHz, d-DMSO): δ12.11(s, 1H), 7.86-7.82(m, 2H), 7.41-7.38(m, 2H), 6.96-6.90(m, 2H), 5.64(s, 2H), 5.11-5.08(m, 3H), 4.83-4.79(m, 4H), 4.66-4.63(m, 4H), 4.56-4.49(m, 8H), 4.25-4.17(m, 5H), 3.84-3.38(m, 32H), 3.31-3.22(m, 10H), 2.47-2.41(m, 4H), 1.95-1.83(m, NMR results confirmed that the structure of formula (IB1) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IB1).

[0093] Synthesis Examples B2 to B5: Synthesis of Drug Conjugates of Formulae (IB2) to (IB5)

[0094] In Synthesis Examples B2 to B11, the drug conjugates of formula (IB2) to (IB11) were synthesized using the same organic synthesis techniques as those described in Synthesis Example B1, and the details of the synthesis are omitted here. NMR and ESI-MS analyses were performed to prove that the drug conjugates of formula (IB2) to (IB11) were successfully obtained.

[0095] The NMR analysis of the resulting drug complex of formula (IB2) is as follows: 1H-NMR (400 MHz, CD3OD): δ8.21(s, 2H), 7.99-7.89(m, 2H), 7.14-7.10(m, 2H), 6.94-6.90(m, 2H), 5.16-5.07(m, 4H), 4.91-4.88(m, 8H), 4.65-4.53(m, 4H), 4.48-4.35(m, 4H), 4.29-4.16(m, 4H), 3.96-3.39(m, 16H), 2.93-2.62(m, 10H), 2.48-2.26(m, 6H), 2.16-1.99 (m, 4H), 1.91-1.54(m, NMR results confirmed that the structure of formula (IB2) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IB2).

[0096] The NMR analysis of the resulting drug complex of formula (IB3) is as follows: 1 H-NMR (400 MHz, D6-DMSO): δ11.30(s, 2H), 8.22-8.05(m, 3H), 7.90-7.81(m, 2H), 7.36-7.28(m, 1H), 7.14-7.05(m, 2H), 6.97-6.80(m, 2H), 4.96-4.88(m, 2H), 4.76-4.58(m, 8H), 4.56-4.33(m, 6H), 4.18-4.03(m, 2H), 3.71-3.61(m, 2H), 3.58-3.48(m, 3H), 3.48-3.34(m, 7H), 3.31-3.15(m, NMR results confirmed that the structure of formula (IB3) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IB3).

[0097] The NMR analysis of the resulting drug complex of formula (IB4) is as follows: 1H-NMR (400 MHz, D6-DMSO): δ11.30(s, 2H), 8.24-8.12(m, 3H), 7.90-7.80(m, 2H), 7.37-7.30(m, 1H), 7.14-7.05(m, 2H), 6.97-6.81(m, 2H), 5.20-5.05(m, 4H), 4.94-4.76(m, 6H), 4.71-4.56(m, 10H), 4.55-4.41(m, 4H), 4.40-4.28(m, 4H), 4.24-4.02(m, 4H), 3.84-3.73(m, NMR confirmed that the structure of formula (IB4) was obtained. ESI-MS also confirmed the presence of peaks corresponding to the molecular weight of the structure of formula (IB4).

[0098] The NMR analysis of the resulting drug complex of formula (IB5) is as follows: 1 H-NMR (400 MHz, DMSO): δ13.87(s, 2H), 8.21(s, 2H), 7.58-7.52(m, 4H), 7.08-7.05(m, 2H), 6.89-6.84(m, 2H), 5.11-5.07(m, 4H), 4.67-4.65(m, 4H), 4.30-4.10(m, 12H), 3.75-3.23(m, 42H), 3.05-2.93(m, 10H), 1.80-1.70(m, 4H), 1.51-1.47(m, 4H), 1.35-1.30(m, 4H), 1.26-1.20(m, 4H). NMR results confirmed that the structure of formula (IB5) was obtained. ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (IB5).

[0099] The NMR analysis of the resulting drug complex of formula (IB6) is as follows: 1H-NMR (400 MHz, CD3OD): δ7.86-7.81(m, 2H), 7.45-7.41(m, 2H), 6.88-6.81(m, 2H), 4.60-4.53(m, 6H), 4.47-4.42(m, 6H), 4.36-4.33(m, 4H), 4.24-4.21(m, 2H), 4.08-4.00(m, 2H), 3.84-3.66(m, 20H), 3.59-3.45(m, 20H), 3.24-3.19(m, 2H), 7.45-7.41(m, 2H), 2.30-2.25(m, NMR confirmed that the structure of formula (IB6) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IB6).

[0100] The NMR analysis of the resulting drug complex of formula (IB7) is as follows: 1 H-NMR (400 MHz, CD3OD): δ7.75-7.71(m, 1H), 7.54-7.51(m, 1H), 7.08 (s, 1H), 7.07-6.99(m, 1H), 6.85-6.75(m, 1H), 4.53-4.26(m, 5H), 3.96-3.68(m, 6H), 3.60-3.36(m, 7H), 3.27-3.00(m, 4H), 2.39-2.34(m, 2H), 2.19-2.12(m, 1H), 2.00-1.87(m, 2H), 1.75-1.66(m, 2H), 1.54-1.47(m, 2H). NMR results confirmed that the structure of formula (IB7) was obtained. ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (IB7).

[0101] The NMR analysis of the resulting drug complex of formula (IB8) is as follows: 1H-NMR (400 MHz, CDCl3): δ8.74-8.72(m, 2H), 8.44-8.41(m, 2H), 7.47-7.43(m, 2H), 5.37-5.28(m, 2H), 5.03-4.95(m, 4H), 4.33-4.23(m, 2H), 4.07-4.03(m, 2H), 3.99-3.93(m, 2H), 3.72-3.65(m, 2H), 3.46-3.38(m, 2H), 2.92-2.85(m, 4H), 2.28-2.22 (m, 2H), 2.09-2.01(m, 24H), 1.72-1.50 (m, 28H). NMR confirmed that the structure of formula (IB8) was obtained. ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (IB8).

[0102] The NMR analysis of the resulting drug complex of formula (IB9) is as follows: 1 H-NMR (400 MHz, CD3OD): δ 7.86-7.80 (m, 2H), 7.35-7.31 (m, 2H), 6.87-6.80 (m, 2H), 4.56-4.52 (m, 2H), 4.45-.4.40 (m, 2H), 3.88-3.37 (m, 22H), 3.31-3.17 (m, 8H), 2.49-2.40 (m, 2H), 2.11-1.52 (m, 20H), 1.32-1.24 (m, 2H). MR results confirmed that the structure of formula (IB9) was obtained. ESI-MS also confirmed peaks corresponding to the molecular weight of the structure of formula (IB9).

[0103] The NMR analysis of the resulting drug complex of formula (IB10) is as follows: 1H-NMR (400 MHz, CD3OD): δ7.57-7.52(m, 2H), 7.44-7.41(m, 2H), 6.91-6.85(m, 2H), 7.57-7.52(m, 2H), 5.40-5.28(m, 4H), 5.03-4.94(m, 4H), 4.33-4.23(m, 8H), 4.12-3.78(m, 6H), 3.82-3.61(m, 2H), 3.50-3.26(m, 2H), 2.25-2.00(m, 24H), 1.87-1.62(m, 16H), 1.48-1.46(m, 10H). The NMR results confirmed that the structure of formula (IB10) was obtained. ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (IB10).

[0104] The NMR analysis of the resulting drug complex of formula (IB11) is as follows: 1 H-NMR (400 MHz, CD3OD): δ8.04-8.00 (m, 2H), 7.86-7.80(m, 1H), 7.38-7.31(m, 1H), 7.12-7.08(m, 2H), 6.88-6.84(m, 2H), 5.04-4.91(m, 4H), 4.73-4.42(m, 16H), 4.30-4.19(m, 2H), 3.81-3.37(m, 16H), 8.04-8.00(m, 2H), 3.22-2.91(m, 6H), 2.05-1.86(m, 4H), 1.71-1.24(m, 6H). The NMR results confirmed that the structure of formula (IB11) was obtained. ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (IB11).

[0105] Synthesis Example C1: Synthesis of a drug conjugate of formula (IC1)

[0106] The drug conjugate of formula (IC1) was synthesized according to the scheme shown in Figure 3. Specifically, to a solution of (R)-1-(6-amino-9H-purin-9-yl)propan-2-ol (300.00 mg, 1.55 mmol) in dry DMF (7 mL), 4-nitrophenyl chloroformate (350.71 mg, 1.74 mmol) and DIPEA (301.15 mg, 2.33 mmol) were added. The reaction mixture was stirred at room temperature for 18 h. The reaction mixture was diluted with EtOAc and washed with 10% LiCl (aq), H2O and NH4Cl (aq). The combined organic layers were dried over MgSO4, filtered and concentrated. The resulting crude product compound 67 was used in the next step without purification.

[0107] To a solution of compound 67 ((R)-1-(6-amino-9H-purin-9-yl)propan-2-yl(4-nitrophenyl)carbonate) (1.86 g, 5.18 mmol) in dry THF (30 mL), propargylamine (712.74 mg, 12.94 mmol) and DIPEA (669.52 mg, 5.18 mmol) were added and stirred at room temperature for 18 h. After removing the solvent, the resulting material was purified by silica gel column chromatography (0-5% MeOH in EtOAc as eluent) to give compound 68 ((R)-1-(6-amino-9H-purin-9-yl)propan-2-ylprop-2-yn-1-ylcarbamate) (707.00 mg, 2.58 mmol, 49.8%).

[0108] To a solution of compound 68 ((R)-1-(6-amino-9H-purin-9-yl)propan-2-ylprop-2-yn-1-ylcarbamate) (174.44 mg, 0.64 mmol) and compound 15 ((2R,3S,4S,5S,6R)-2-((6-azidohexyl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol) (161.83 mg, 0.53 mmol) in HO (2.5 mL) was added a solution of L-ascorbic acid (36.99 mg, 0.21 mmol) and CuSO45HO (26.25 mg, 0.11 mmol) in HO (2.5 mL). The reaction mixture was stirred at room temperature for 1-2 h. The resulting material was purified by C18 column chromatography (0-12% ACN in HO as eluent) to give compound 105 ((R)-1-(6-amino-9H-purin-9-yl)propan-2-yl ((1-(6-(((2R,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxylmethyl)tetrahydro-2H-pyran-2-yl)oxy)hexyl)-1H-1,2,3-triazol-4-yl)methyl)carbamate) (152.30 mg, 0.26 mmol, 49.1% yield). Compound 105 corresponds to the structure of formula (IC1).

[0109] The NMR analysis of compound 105 (formula (IC1)) is as follows: 1 H-NMR (400 MHz, DO): δ 8.12 (s, 1H), 8.10 (s, 1H), 7.50 (s, 1H), 5.17-5.14 (m, 1H), 4.38-4.03 (m, 7H), 3.90-3.43 (m, 8H), 1.83-1.74 (m, 2H), 1.49-1.16 (m, 9H). NMR confirmed that the structure of formula (IC1) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IC1).

[0110] Synthesis Examples C2 to C11: Synthesis of Drug Conjugates of Formulae (IC2) to (IC11)

[0111] In Synthesis Examples C2 to C11, the drug conjugates of formula (IC2) to (IC11) were synthesized using the same organic synthesis techniques as those described in Synthesis Example C1, and the details of the synthesis are omitted here. NMR and ESI-MS analyses were performed to prove that the drug conjugates of formula (IC2) to (IC11) were successfully obtained.

[0112] The NMR analysis of the resulting drug complex of formula (IC2) is as follows: 1 H-NMR (400 MHz, D2O): δ8.20(s, 1H), 8.15(s, 1H), 7.56(s, 1H), 5.23-5.19(m, 1H), 4.47-4.09(m, 7H), 3.97-3.86(m, 2H), 3.78-3.74(m, 1H), 3.67-3.61(m, 1H), 3.55-3.40(m, 3H), 3.31-3.27(m, 1H), 1.89-1.80(m, 2H), 1.58-1.21(m, 9H), 1 H-NMR (400 MHz, d6-DMSO): δ8.13(s, 1H), 8.06(s, 1H), 7.77(s, 1H), 7.65(t, 1H), 7.19(s, 2H), 5.08-5.00(m, 1H), 4.95-4.88(m, 3H), 4.48-4.46(m, 1H), 4.32-4.26(m, 4H), 4.16-4.08(m, 3H), 3.76-3.64(m, 2H), 3.15-3.00(m, 4H), 2.94-2.90(m, 1H), 1.77(p, 2H), 1.48(p, 2H), 1.32(p, 2H), 1.22(p, 2H), 1.13(d, 3H). NMR confirmed that the structure of formula (IC2) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IC2).

[0113] The NMR analysis of the resulting drug complex of formula (IC3) is as follows: 1H-NMR (400 MHz, d6-DMSO): δ8.13(s, 1H), 8.03(s, 1H), 7.17(s, 2H), 7.08(t, 1H), 5.12-5.10(m, 2H), 5.04-4.98(m, 1H), 4.81(s, br, 1H), 4.66(s, br, 2H), 4.57-4.52(m, 2H), 4.30-4.16(m, 4H), 3.77-3.73(m, 2H), 3.61-3.40(m, 8H), 3.03-2.97(m, 1H), 2.88(q, 2H), 1.51-1.46(m, 2H), 1.34-1.21(m, 6H), 1.12(d, 3H). NMR confirmed that the structure of formula (IC3) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IC3).

[0114] The NMR analysis of the resulting drug complex of formula (IC4) is as follows: 1 H-NMR (400 MHz, DO): δ 8.15 (s, 1H), 8.12 (s, 1H), 7.51 (s, 1H), 5.19-5.15 (m, 1H), 4.47-3.93 (m, 10H), 3.87-3.53 (m, 11H), 3.32-3.27 (m, 1H), 1.85-1.74 (m, 2H), 1.56-1.52 (m, 2H), 1.37-1.18 (m, 7H). NMR confirmed that the structure of formula (IC4) was obtained. ESI-MS also confirmed peaks corresponding to the molecular weight of the structure of formula (IC4).

[0115] The NMR analysis of the resulting drug complex of formula (IC5) is as follows: 1H-NMR (400 MHz, DO): δ 8.19 (s, 1H), 8.15 (s, 1H), 7.55 (s, 1H), 5.22-5.19 (m, 1H), 4.47-4.09 (m, 7H), 3.97-3.76 (m, 4H), 3.72-3.60 (m, 3H), 3.56-3.52 (m, 1H), 1.88-1.73 (m, 2H), 1.60-1.56 (m, 2H), 1.40-1.21 (m, 7H). NMR confirmed that the structure of formula (IC5) was obtained. ESI-MS also confirmed peaks corresponding to the molecular weight of the structure of formula (IC5).

[0116] The NMR analysis of the resulting drug complex of formula (IC6) is as follows: 1 H-NMR (400 MHz, D2O): δ8.08 (s, 1H), 8.05(s, 1H), 7.54(s, 1H), 5.14-5.05(m, 1H), 4.43-4.17(m, 9H), 4.16-3.97(m, 3H), 3.95-3.83(m, 5H), 3.82-3.63(m, 12H), 3.63-3.44(m, 10H), 3.28-3.19(m, 2H), 3.18-2.97(m, 5H), 2.23-2.16(m, 4H), 2.15-1.91(m, 4H), 1.91-1.78(m, NMR confirmed that the structure of formula (IC6) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IC6).

[0117] The NMR analysis of the resulting drug complex of formula (IC7) is as follows: 1H-NMR (400 MHz, DO): δ 8.11 (s, 1H), 8.07 (s, 1H), 7.58 (s, 1H), 5.14-5.05 (m, 1H), 4.44-4.00 (m, 15H), 3.97-3.42 (m, 47H), 3.28-3.18 (m, 3H), 3.14-2.99 (m, 7H), 2.45-2.34 (m, 7H), 2.18-1.99 (m, 5H), 1.58-1.13 (m, 32H). NMR confirmed that the structure of formula (IC7) was obtained. ESI-MS also confirmed peaks corresponding to the molecular weight of the structure of formula (IC7).

[0118] The NMR analysis of the resulting drug complex of formula (IC8) is as follows: 1 H-NMR (400 MHz, DO): δ 8.16(s, 1H), 8.11(s, 1H), 7.51(s, 1H), 5.41-5.38(m, 2H), 5.19-5.16(m, 1H), 4.43-4.17(m, 6H), 4.08-3.40(m, 22H), 3.30-3.25(m, 1H), 1.85-1.75(m, 2H), 1.55-1.52(m, 2H), 1.37-1.16(m, 7H). NMR confirmed that the structure of formula (IC8) was obtained. ESI-MS also confirmed peaks corresponding to the molecular weight of the structure of formula (IC8).

[0119] The NMR analysis of the resulting drug complex of formula (IC9) is as follows: 1 H-NMR (400 MHz, DO): δ 8.15 (s, 1H), 8.12 (s, 1H), 7.50 (s, 1H), 5.18-5.14 (m, 1H), 4.46-4.04 (m, 7H), 3.93-3.42 (m, 8H), 1.97 (s, 3H), 1.84-1.74 (m, 2H), 1.44-1.14 (m, 9H). NMR confirmed that the structure of formula (IC9) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IC9).

[0120] The NMR analysis of the resulting drug complex of formula (IC10) is as follows:1 H-NMR (400 MHz, DO): δ 8.18-7.98 (m, 2H), 7.71-7.43 (m, 1H), 7.28-7.16 (m, 2H), 7.11-6.95 (m, 2H), 5.58-5.39 (m, 1H), 5.19-4.92 (m, 2H), 4.51-4.47 (d, 1H, J = 7.8 Hz), 4.41-3.91 (m, 5H), 3.90-3.65 (m, 10H), 3.65-3.53 (m, 2H), 1.37-1.27 (d, 3H, J = 6.3 Hz). NMR results confirmed the structure of formula (IC10). ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (IC10).

[0121] The NMR analysis of the resulting drug complex of formula (IC11) is as follows: 1 H-NMR (400 MHz, DO): δ 8.14-7.97 (m, 2H), 7.03-6.79 (m, 5H), 5.19-5.06 (m, 1H), 5.06-4.99 (d, 1H, J = 7.8), 4.45-4.15 (m, 3H), 4.08-3.61 (m, 11H), 3.61-3.44 (m, 3H), 1.33-1.24 (d, 3H, J = 6.2 Hz). NMR confirmed that the structure of formula (IC11) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IC11).

[0122] Synthesis Example D1: Synthesis of a drug conjugate of formula (ID1)

[0123] The drug conjugate of formula (ID1) was synthesized according to the scheme shown in Figure 4. Specifically, to a solution of compound 3 ((3R,4S,5R,6R)-6-(acetoxymethyl)-5-(((2S,3R,4S,5S,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2,3,4-triyl triacetate) (6.87 g, 10.1 mmol), compound 22 (benzyl 6-hydroxyhexanoate) (4.50 g, 20.02 mmol) and 4 Å MS (1.00 g) in DCM (70 mL) was added BFOEt (5.1 mL, 4.05 mmol) at 0°C, stirred for 15 minutes, and then warmed to room temperature over 12 hours. The mixture was quenched with NaHCO3(aq) and extracted with DCM. The combined organics were dried over MgSO4 and concentrated. The residue was purified by silica gel column chromatography using EtOAc:Hexane (1:2) as eluent to give compound 23 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-(benzyloxy)-6-oxohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (2.52 g, 3.00 mmol, 30% yield).

[0124] To a solution of compound 23 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-(benzyloxy)-6-oxohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (600 mg, 0.71 mmol) in MeOH (5 mL) was added 10% palladium on carbon (60 mg, 0.1 w%) and stirred under H2 for 3 h. The mixture was filtered through Celite to remove palladium on carbon, concentrated under reduced pressure and the solvent was removed by evaporation. The crude product compound 24 was used in the next step without purification.

[0125] Compound 24 (6-(((2R,3R,4S,5R,6R)-3,4-diacetoxy-6-(acetoxymethyl)-5-(((2S,3R,4S,5S,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexanoic acid) (448 mg, 0.60 mmol) and (S)-(4-fluorophenyl)(2-(pyrrolidin-2-yl)thiazol-4-yl)methanoneine (200 mg, 0.64 mmol) in ACN (10 mL) were added with HATU (450 mg, 1.20 mmol) and NMM (350 μL, 3.19 mmol) and stirred at room temperature for 12 hours. The mixture was concentrated and purified by silica gel column chromatography to give the product compound 123 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-6-oxohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (435 mg, 0.43 mmol, 72% yield).

[0126] To a solution of compound 123 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-6-oxohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (435 mg, 0.43 mmol) in MeOH (20 mL), NaOMe (330 mg, 6.11 mmol) was added at 0° C. After the addition, the reaction mixture was allowed to warm to room temperature and stirred for 1 h. The reaction mixture was then neutralized with Amberlite IR-120(H) until pH 7. The solution was then filtered and the solvent was removed. The resulting material was purified by reverse phase silica gel column chromatography to give the product compound 124 (6-(((2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-1-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)hexan-1-one) (200 mg, 0.28 mmol, 65% yield). Compound 124 corresponds to the structure of formula (ID1).

[0127] The NMR analysis of compound 105 (formula (ID1)) is as follows: 1H-NMR (400 MHz, CD3OD): δ 8.34-8.32 (m, 1H), 8.26-8.23 (m, 2H), 7.28-7.24 (m, 2H), 5.54-5.45 (m, 2H), 4.37-4.21 (m, 3H), 3.92-3.66 (m, 10H), 3.60-3.37 (m, 8H), 3.25-3.13 (m, 2H), 2.51-2.31 (m, 4H), 2.21-2.04 (m, 2H), 1.70-1.64 (m, 4H), 1.53-1.47 (m, 2H). NMR confirmed that the structure of formula (ID1) was obtained. ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (ID1).

[0128] Synthesis Examples D2 to D5: Synthesis of Drug Conjugates of Formulae (ID2) to (ID5)

[0129] In Synthesis Examples D2 to D5, the drug conjugates of formulae (ID2) to (ID5) were synthesized using the same organic synthesis techniques as those described in Synthesis Example D1, and the details of the synthesis are omitted here. NMR and ESI-MS analyses were performed to prove that the drug conjugates of formulae (ID2) to (ID5) were successfully obtained.

[0130] The NMR analysis of the resulting drug complex of formula (ID2) is as follows: 1H-NMR (400 MHz, D2O): δ8.22(s, 1H), 7.89-7.78(m, 2H), 7.24-7.15(m, 2H), 5.26-5.15(m, 1H), 4.90-4.81 (m, 1H), 4.488 (d, J=7.24, 1H), 4.387(d, J=8, 1H), 4.19-4.09(m, 2H), 4.04-3.95 (m, H), 3.94-3.81(m, 3H), 3.79-3.42(m, 11H), 3.32-3.23(m, 1H), 2.51-2.30(m, 1H), 2.12-1.89(m, NMR results confirmed that the structure of formula (ID2) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (ID2).

[0131] The NMR analysis of the resulting drug complex of formula (ID3) is as follows: 1 H-NMR (400 MHz, D2O): δ8.20(s, 1H), 7.95-7.91(m, 2H), 7.78(s, 1H), 7.35-7.31(m, 2H), 5.40-5.35(m, 2H), 5.27-5.24(m, 2H), 4.93-4.90(m, 1H), 4.41-4.39(m, 2H), 4.25-4.22(m, 2H), 3.98-3.41(m, 29H), δ3.30-3.25(m, 1H), 2.54-2.45(m, 1H), 2.12-2.02(m, 4H), 1.87-1.83(m, NMR confirmed that the structure of formula (ID3) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (ID3).

[0132] The NMR analysis of the resulting drug complex of formula (ID4) is as follows: 1H-NMR (400 MHz, D2O): δ8.20(s, 1H), 8.15(s, 1H), 7.98(s, 1H), 7.96-7.83(m, 3H), 7.78(s, 1H), 7.27(m, 2H), 5.31-5.16(m, 3H), 4.90-4.84(m, 1H), 4.42-4.34(m, 6H), 4.28-4.20(m, 2H), 4.12-4.03(m, 2H), 3.92-3.83(m, 5H), 3.82-3.62(m, 7H), 3.61-3.44(m, 10H), 3.25-3.18(m, NMR results confirmed that the structure of formula (ID4) was obtained. ESI-MS also confirmed peaks corresponding to the molecular weight of the structure of formula (ID4).

[0133] The NMR analysis of the resulting drug complex of formula (ID5) is as follows: 1 H-NMR (400 MHz, DO): δ 8.25 (s, 1H), 8.05-8.01 (m, 2H), 7.30-7.25 (m, 2H), 5.44-5.41 (m, 1H), 4.90-4.89 (m, 1H), 4.46-4.44 (m, 1H), 3.94-3.53 (m, 18H), 8.34-8.32 (m, 1H), 2.43-2.06 (m, 7H), 1.64-1.09 (m, 18H). NMR confirmed that the structure of formula (ID5) was obtained. ESI-MS also confirmed the presence of peaks corresponding to the molecular weight of the structure of formula (ID5).

[0134] Synthesis Example E1: Synthesis of a drug conjugate of formula (IE1)

[0135] The drug conjugate of formula (IE1) was synthesized according to the scheme shown in Figure 5. Specifically, a solution of 1-((2S,4R,5S)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (300.37 mg, 1.24 mmol) and imidazole (168.83 mg, 2.48 mmol) in dry DMF (0.6 mL) was added to a solution of TBSCl (186.89 mg, 1.24 mmol) in dry DMF (0.6 mL) at 0 °C and stirred for 30 min. It was then warmed to room temperature and stirred for 18 h. The reaction mixture was diluted with EtOAc and washed with 10% LiCl (aq) and NaHCO3 (aq). The combined organic layers were dried over MgSO4, filtered and concentrated to give compound 60 (1-((2S,4R,5S)-5-(((tert-butyldimethylsilyl)oxy)methyl)-4-hydroxytetrahydrofuran-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione) (377.10 mg, 1.06 mmol, 85.4% yield).

[0136] To a solution of compound 60 (1-((2S,4R,5S)-5-(((tert-butyldimethylsilyl)oxy)methyl)-4-hydroxytetrahydrofuran-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione) (950.00 mg, 2.66 mmol) in dry DCM (26 mL) was added DMAP (422.12 mg, 3.46 mmol) and Ac2O (407.34 mg, 3.99 mmol). The mixture was stirred at room temperature for 6 h. After removal of the solvent, the resulting material was dissolved in EtOAc and washed with NH4Cl(aq). The combined organic layers were dried over MgSO4, filtered and concentrated to give compound 61 ((2S,3R,5S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl acetate) (1060.09 mg, 2.66 mmol, yield equiv.).

[0137] To a solution of compound 61 ((2S,3R,5S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl acetate) (530.05 mg, 1.33 mmol) in THF (9 mL), a 1M THF solution (2 mL) of TBAF was added at 0° C. and stirred for 1.5 hours. After removal of the solvent, the resulting material was purified by silica gel column chromatography (40% hexane in acetone as eluent) to give compound 62 ((2S,3R,5S)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl acetate) (266.40 mg, 0.94 mmol, 70.6%).

[0138] To a solution of compound 62 ((2S,3R,5S)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl acetate) (310.00 mg, 1.09 mmol) in dry DCM (7 mL) was added 4-nitrophenyl chloroformate (330.56 mg, 1.64 mmol) and DIPEA (351.56 mg, 2.72 mmol). The reaction mixture was stirred at room temperature for 2 h. After removal of the solvent, the resulting material was dissolved in EtOAc and washed with H2O and NH4Cl (aq). The combined organic layers were dried over MgSO4, filtered and concentrated. The resulting crude product compound 63 was used in the next step without purification.

[0139] To a solution of compound 63 ((2S,3R,5S)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-((((4-nitrophenoxy)carbonyl)oxy)methyl)tetrahydrofuran-3-yl acetate) (489.80 mg, 1.09 mmol) in dry THF (7 mL) was added compound 5 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-aminohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (1.16 g, 1.58 (351.56 mg, 2.72 mmol) and DIPEA (351.56 mg, 2.72 mmol) were added. The reaction mixture was heated to 70° C. and stirred for 18 h. After removal of the solvent, the mixture was dissolved in EtOAc and washed with H2O and NH4Cl (aq). The combined organic layers were dried over MgSO4, filtered and concentrated. The resulting material was purified by silica gel column chromatography (14.3% hexanes in EtOAc as eluent) to give compound 97 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-6-((6-(((((2S,3R,5S)-3-acetoxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl)methoxy)carbonyl)amino)hexyl)oxy)-2-(acetoxymethyl)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (474.60 mg, 0.45 mmol, 41.3%).

[0140] To a solution of compound 97 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-6-((6-(((((2S,3R,5S)-3-acetoxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl)methoxy)carbonyl)amino)hexyl)oxy)-2-(acetoxymethyl)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (210.00 mg, 0.20 mmol) in MeOH (2 mL), NaOMe (27.01 mg, 0.50 mmol) was added and stirred at room temperature for 2 hours. The reaction mixture was then neutralized with resin until the pH was 7. The solution was then filtered and the solvent was removed. The resulting material was purified by C18 column chromatography (0-20% MeOH in HO as eluent) to give compound 98 (((2S,3R,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl)methyl (6-(((2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-(((2S,3R,4S,-5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexyl)carbamate) (51.30 mg, 0.07 mmol, 35.0% yield). Compound 98 corresponds to the structure of formula (IE1).

[0141] The NMR analysis results of compound 98 (formula (IE1)) are as follows: 1H-NMR (400 MHz, DO): δ 7.51 (s, 1H), 6.28-6.24 (t, 1H), 4.53-4.41 (m, 4H), 4.29-4.17 (m, 2H), 3.99-3.54 (m, 13H), 3.33-3.29 (m, 1H), 3.12-3.09 (t, 2H), 2.46-2.31 (m, 2H), 1.89 (s, 3H), 1.60-1.32 (m, 8H). NMR confirmed that the structure of formula (IE1) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IE1).

[0142] Synthesis Example F1: Synthesis of a drug conjugate of formula (IF1)

[0143] The drug conjugate of formula (IF1) was synthesized according to the scheme shown in FIG. 6. Specifically, to a solution of 2-(6-amino-9H-purin-9-yl)ethan-1-ol (500.00 mg, 2.79 mmol) in dry DMF (5.5 mL), 4-nitrophenyl chloroformate (1.52 g, 7.54 mmol) and DIPEA (1.08 g, 8.37 mmol) were added. The reaction mixture was stirred at room temperature under N2 for 18 h, resulting in a yellow solution containing a precipitate. The reaction mixture was then diluted with H2O, and the resulting precipitate was collected as a pale yellow solid (compound 58). The pale yellow solid was then dissolved in dry THF (18 mL), and propargylamine (384.46 mg, 6.98 mmol) and DIPEA (360.61 mg, 2.79 mmol) were added. The reaction mixture was stirred at room temperature under N2 for 4 h, resulting in an orange solution containing a precipitate. The solvent was removed under reduced pressure and the precipitate was washed with EtOAc and acetone to give compound 59 (2-(6-amino-9H-purin-9-yl)ethylprop-2-yn-1-ylcarbamate) (314.90 mg, 1.21 mmol, 43.4% yield).

[0144] To a solution of compound 59 (2-(6-amino-9H-purin-9-yl)ethylprop-2-yn-1-ylcarbamate) (83.28 mg, 0.32 mmol) and compound 6 ((2S,3R,4S,5R,6R)-2-(((2R,3S,4R,5R,6R)-6-((6-azidohexyl)oxy)-4,5-dihydroxy-2-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol) (122.90 mg, 0.26 mmol) in MeOH (2 mL) was added a green solution of L-ascorbic acid (18.65 mg, 0.11 mmol) and CuSO4·5H2O (13.25 mg, 0.05 mmol) in HO (2 mL). The reaction mixture was stirred at room temperature for 1-2 h to give a yellow-green solution. MeOH was removed and the resulting material was purified by C18 column chromatography (0-15% ACN in HO as eluent) to give compound 96 (2-(6-amino-9H-purin-9-yl)ethyl ((1-(6-(((2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxylmethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexyl)-1H-1,2,-3-triazol-4-yl)methyl)carbamate) (70.70 mg, 0.10 mmol, 38.5% yield). Compound 96 corresponds to the structure of formula (IF1).

[0145] The NMR analysis results of compound 96 (formula (IF1)) are as follows: 1H-NMR (400 MHz, DO): δ 8.19 (s, 1H), 8.17 (s, 1H), 7.66 (s, 1H), 4.53-4.44 (m, 7H), 4.28-4.23 (m, 3H), 4.02-3.97 (m, 2H), 3.91-3.57 (m, 12H), 3.36-3.32 (m, 1H), 1.90-1.79 (m, 2H), 1.61-1.57 (m, 2H), 1.35-1.18 (m, 4H). NMR confirmed that the structure of formula (IF1) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IF1).

[0146] Synthesis Example F2: Synthesis of a drug conjugate of formula (IF2)

[0147] In Synthesis Example F2, the drug conjugate of formula (IF2) was synthesized using the same organic synthesis techniques as those described in Synthesis Example F1, and the details of the synthesis are omitted here. NMR and ESI-MS analyses were performed to prove that the drug conjugate of formula (IF2) was successfully obtained.

[0148] The NMR analysis of the drug complex of formula (IF2) is as follows: 1 H-NMR (400 MHz, DO): δ 8.23(s, 1H), 8.16(s, 1H), 4.48-4.46(m, 5H), 4.00-3.54(m, 15H), 2.88-2.85(m, 2H), 1.58-1.55(m, 2H), 1.27-1.07(m, 6H). NMR confirmed that the structure of formula (IF2) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IF2).

[0149] Synthesis Example G1: Synthesis of drug conjugate of formula (IG1)

[0150] The drug conjugate of formula (IG1) was synthesized according to the scheme shown in Figure 7. Specifically, to a solution of lamivudine (2.00 g, 8.72 mmol) in ACN (90 mL), Ac2O (1 mL, 8.99 mmol) was added and stirred at room temperature for 12 h. The reaction mixture was then diluted with HO and the resulting precipitate was collected. The crude product, compound 64 (N-(1-((2R,5S)-2-(hydroxymethyl)-1,3-oxathiolan-5-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide) (2.03 g, crude yield 85%), was used in the next step without purification.

[0151] To a solution of compound 64 (N-(1-((2R,5S)-2-(hydroxymethyl)-1,3-oxathiolan-5-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide) (1.00 mg, 3.69 mmol) in THF (40 mL), CDI (1.20 g, 7.37 mmol) was added and stirred at 50° C. for 3 h. Then, prop-2-yn-1-amine (0.90 mL, 14.74 mmol) was added and stirred at 50° C. for 12 h. The solvent was removed under reduced pressure. The resulting material was purified by silica gel column chromatography using EtOAc:MeOH (5%) as the eluent to give the product compound 65 (((2R,5S)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-1,3-oxathiolan-2-yl)methylprop-2-yn-1-ylcarbamate) (0.72 g, 2.04 mmol, 55% yield).

[0152] To a solution of compound 65 (((2R,5S)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-1,3-oxathiolan-2-yl)methylprop-2-yn-1-ylcarbamate) (260 mg, 0.72 mmol) in MeOH (5 mL) was added NaOMe (40 mg, 0.72 mmol) at 0° C. After the addition, the reaction mixture was allowed to warm to room temperature and stirred for 15 min. The reaction mixture was then neutralized with Amberlite IR-120(H) until pH 7. The solution was then filtered and the solvent was removed. The crude product compound 66 was used in the next step without purification.

[0153] A solution of compound 66 (((2R,5S)-5-(4-amino-2-oxopyrimidin-1(2H)-yl)-1,3-oxathiolan-2-yl)methylprop-2-yn-1-ylcarbamate) (223 mg, 0.72 mmol) and compound 6 ((2S,3R,4S,5R,6R)-2-(((2R,3S,4R,5R,6R)-6-((6-azidohexyl)oxy)-4,5-dihydroxy-2-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol) (280 mg, 0.60 mmol) in THF (3 mL) was added with CuSO4·H2O (30 mg, 0.12 mmol) and ascorbic acid (42 mg, 0.24 mmol). A solution of 1 mmol) in H2O (3 mL) was added and stirred at room temperature for 3 h. The mixture was concentrated under reduced pressure to remove THF. The resulting material was purified by reverse-phase silica gel column chromatography using ACN:HO (15%) as eluent to give the product compound 99 (((2R,5S)-5-(4-amino-2-oxopyrimidin-1(2H)-yl)-1,3-oxathiolan-2-yl)methyl ((1-(6-(((2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexyl)-1H-1,2,3-triazol-4-yl)methyl)carbamate) (300 mg, 0.39 mmol, 64% yield). Compound 99 corresponds to the structure of formula (IG1).

[0154] The NMR analysis of compound 99 (formula (IG1)) is as follows: 1H-NMR (400 MHz, CD3OD): δ7.85-7.83(m, 1H), 7.82 (s, 1H), 6.29-6.27(m, 1H), 5.88-5.87(m, 1H), 5.40-5.39(m, 1H), 4.58(s, 1H), 4.54-4.34(m, 7H), 4.29-4.27(m, 1H), 3.91-3.68(m, 7H), 3.60-3.47(m, 7H), 3.41-3.38(m, 1H), 3.26-3.22(m, 1H), 3.14-3.10(m, 1H), 1.90-1.83(m, NMR results confirmed that the structure of formula (IG1) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IG1).

[0155] Synthesis Example H1: Synthesis of a drug conjugate of formula (IH1)

[0156] The drug conjugate of formula (IH1) was synthesized according to the scheme shown in Figure 8. Specifically, TFA (3 mL, 39.18 mmol) was added to a solution of tert-butyl ((S)-1-cyclohexyl-2-oxo-2-((S)-2-((4-phenyl-1,2,3-thiadiazol-5-yl)carbamoyl)pyrrolidin-1-yl)ethyl)carbamate (500 mg, 0.97 mmol) in DCM (3 mL) and stirred at room temperature. After the reaction was completed, the solvent and TFA were removed under reduced pressure, and the crude product compound 70 ((S)-1-((S)-2-amino-2-cyclohexylacetyl)-N-(4-phenyl-1,2,3-thiadiazol-5-yl)pyrrolidine-2-carboxamide trifluoroacetate) (equivalent yield) was used in the next step without purification.

[0157] A solution of compound 24 (6-(((2R,3R,4S,5R,6R)-3,4-diacetoxy-6-(acetoxymethyl)-5-(((2S,3R,4S,5S,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexanoic acid) (720 mg, 0.96 mmol) and compound 70 ((S)-1-((S)-2-amino-2-cyclohexylacetyl)-N-(4-phenyl-1,2,3-thiadiazol-5-yl)pyrrolidine-2-carboxamide trifluoroacetate) (512 mg, 0.97 mmol) in ACN (10 mL) was added with HATU (370 mg, 0.97 mmol) and NMM (0.5 mL, 4.54 mmol). The mixture was added with 2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-(((S)-1-cyclohexyl-2-oxo-2-((S)-2-((4-phenyl-1,2,3-thiadiazol-5-yl)carbamoyl)pyrrolidin-1-yl)ethyl)amino)-6-oxohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (550 mg, 0.48 mmol, yield 50%).

[0158] To a solution of compound 113 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-(((S)-1-cyclohexyl-2-oxo-2-((S)-2-((4-phenyl-1,2,3-thiadiazol-5-yl)carbamoyl)pyrrolidin-1-yl)ethyl)amino)-6-oxohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (550 mg, 0.48 mmol) in MeOH (10 mL), NaOMe (350 mg, 6.48 mmol) was added at 0° C. After warming to room temperature, the reaction mixture was stirred until the reaction was complete. The mixture was quenched with Amberlite IR-120(H) and filtered off. The resulting material was purified by reverse phase silica gel column chromatography to give the product compound 114 ((S)-1-((S)-2-cyclohexyl-2-(6-(((2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexanamido)acetyl)-N-(4-phenyl-1,2,3-thiadiazol-5-yl)pyrrolidine-2-carboxamide) (180 mg, 0.21 mmol, 44% yield). Compound 114 corresponds to the structure of formula (IH1).

[0159] The NMR analysis result of compound 114 (formula (IH1)) is as follows: 1H-NMR (400 MHz, CD3OD): δ7.76-7.74(m, 2H), 7.61-7.58(m, 2H), 7.54-7.50(m, 1H), 4.58(s, 1H), 4.42-4.26(m, 4H), 4.04-3.99(m, 1H), 3.91-3.67(m, 9H), 3.60-3.46 (m, 7H), 3.40-3.36(m, 1H), 3.25-3.20(m, 1H), 2.25-2.04(m, 7H), 1.85-1.58(m, 10H), 1.44-1.36(m, 2H), 1.24-1.16(m, 2H), 1.10-0.97 (m, 2H). NMR confirmed that the structure of formula (IH1) was obtained. ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (IH1).

[0160] Synthesis Examples H2-H3: Synthesis of Drug Conjugates of Formulae (IH2)-(IH3)

[0161] In Synthesis Examples H2-H3, the drug conjugates of formula (IH2)-formula (IH3) were synthesized using the same organic synthesis techniques as described in Synthesis Example H1, and the details of the synthesis are omitted here. NMR and ESI-MS analyses were performed to prove that the drug conjugates of formula (IH2)-formula (IH3) were successfully obtained.

[0162] The NMR analysis of the drug complex of formula (IH2) is as follows: 1 H-NMR (400 MHz, CD3OD): δ 7.75-7.72 (m, 2H), 7.62-7.57 (m, 2H), 7.55-7.51 (m, 1H), 4.52 (s, 4H), 4.37-4.34 (m, 1H), 4.29-4.27 (m, 1H), 3.91-3.37 (m, 15H), 3.25-3.21 (m, 1H), 2.43-2.39 (m, 2H), 2.21-2.01 (m, 4H), 1.67-1.60 (m, 4H), 1.47-1.39 (m, 2H). NMR results confirmed that the structure of formula (IH2) was obtained. ESI-MS also confirmed a peak corresponding to the molecular weight of the structure of formula (IH2).

[0163] The NMR analysis of the drug complex of formula (IH3) is as follows: 1 H-NMR (400 MHz, D6-DMSO): δ8.16-8.04(m, 1H), 7.95-7.79(br, 2H), 7.62-7.29(br, 3H), 5.22-5.14(m, 1H), 5.09(d, 1H, J=3.6), 4.91-4.80(m, 1H), 4.70-4.57(m, 4H), 2.11-1.77(br, 4H), 4.50(d, 1H, J=4.4), 4.45-4.23(m, 4H), 4.23-4.17(m, 1H), 4.15-3.97(m, 2H), 3.86-3.75(m, NMR results confirmed that the structure of formula (IH3) was obtained. ESI-MS also confirmed the presence of peaks corresponding to the molecular weight of the structure of formula (IH3).

[0164] Synthesis Example J1: Synthesis of a drug conjugate of formula (IJ1)

[0165] The drug conjugate of formula (IJ1) was synthesized according to the scheme shown in Figure 9. Specifically, TFA (3 mL, 39.18 mmol) was added to a solution of tert-butyl ((5S,10aR)-8-(benzhydrylcarbamoyl)-3-(3-methylbutanoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazocin-5-yl)carbamate (200 mg, 0.35 mmol) in DCM (3 mL) and stirred at room temperature. After the reaction was completed, the solvent and TFA were removed under reduced pressure. The crude product compound 74 ((5S,10aR)-5-amino-N-benzhydryl-3-(3-methylbutanoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazocin-8-carboxamide trifluoroacetate) (equivalent yield) was used in the next step without purification.

[0166] A solution of compound 24 (6-(((2R,3R,4S,5R,6R)-3,4-diacetoxy-6-(acetoxymethyl)-5-(((2S,3R,4S,5S,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexanoic acid) (390 mg, 0.52 mmol) and compound 74 ((5S,10aR)-5-amino-N-benzhydryl-3-(3-methylbutanoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazocine-8-carboxamide trifluoroacetate) (207 mg, 0.35 mmol) in ACN (5 mL) was added with HATU (265 mg, 0.69 mmol) and NMM (0.2 mL, 1.86 mmol) was added and stirred at room temperature for 12 hours. The mixture was purified by silica gel column chromatography to obtain compound 118 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-(((5S,10aR)-8-(benzhydrylcarbamoyl)-3-(3-methylbutanoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazocin-5-yl)amino)-6-oxohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (320 mg, 0.26 mmol, yield 75%).

[0167] To a solution of compound 118 ((2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((2R,3R,4S,5R,6R)-4,5-diacetoxy-2-(acetoxymethyl)-6-((6-(((5S,10aR)-8-(benzhydrylcarbamoyl)-3-(3-methylbutanoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazocin-5-yl)amino)-6-oxohexyl)oxy)tetrahydro-2H-pyran-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate) (320 mg, 0.26 mmol) in MeOH (10 mL), NaOMe (200 mg, 3.70 mmol) was added at 0° C. After warming to room temperature, the reaction mixture was stirred until the reaction was complete. The mixture was quenched with Amberlite IR-120(H) and filtered off. The resulting material was purified by reverse phase silica gel column chromatography to give the product compound 119 ((5S,10aR)-N-benzhydryl-5-(6-(((2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)hexanamido)-3-(3-methylbutanoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazocine-8-carboxamide) (90 mg, 0.10 mmol, 38% yield). Compound 118 corresponds to the structure of formula (IJ1).

[0168] The NMR analysis result of compound 118 (formula (IJ1)) is as follows: 1H-NMR (400 MHz, CD3OD): δ-7.25 (m, 10H), 6.14 (s, 1H), 4.83-4.81 (m, 1H), 4.77-4.76 (m, 1H), 4.59 (s, 1H), 4.35-4.33 (m, 1H), 4.23-4.18 (m, 1H), 3.94-3.33 (m, 17H), 2.48-1.40 (m, 21H), 0.98-0.94 (m, 6H). NMR confirmed that the structure of formula (IJ1) was obtained. ESI-MS also confirmed the peaks corresponding to the molecular weight of the structure of formula (IJ1).

[0169] Synthesis Example J2: Synthesis of drug conjugate of formula (IJ2)

[0170] In Synthesis Example J2, the drug conjugate of formula (IJ2) was synthesized using the same organic synthesis technique as described in Synthesis Example J1, and the details of the synthesis are omitted here. NMR and ESI-MS analyses were performed to prove that the drug conjugate of formula (IJ2) was successfully obtained.

[0171] The NMR analysis of the drug complex of formula (IJ2) is as follows: 1 H-NMR (400 MHz, D6-DMSO): δ8.92-8.86(m, 1H), 8.11(s, H), 7.37-7.21(m, 10H), 6.91-6.85(m, 1H), 6.07(d, 1H, J=8.8), 5.19-5.06(m, 2H), 4.84(d, 1H, J=12.0), 4.80-4.76(m, 2H), 4.70-4.60(m, 4H), 4.54-4.41(m, 4H), 4.36-4.30(m, 4H), 4.23-4.18(m, 2H), 4.13-4.04(m, NMR confirmed that the structure of formula (IJ2) was obtained. ESI-MS also confirmed the presence of peaks corresponding to the molecular weight of the structure of formula (IJ2).

[0172] Experimental Example:

[0173] In the following experimental examples, in vitro cell line evaluation and in vitro animal studies were conducted to demonstrate that the drug conjugate of the present invention is superior to conventional methods in treating Hepatitis B. The following materials and methods can be applied to all experimental examples.

[0174] [Cell line and culture conditions] - HepG2 2.2.15 cells In the following experimental examples, when HepG2 2.2.15 cells (a cell line expressing hepatitis B virus) are used, the cells are maintained in Dulbecco's modified Eagle's medium (DMEM) (Gibco BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA), 100 U / mL penicillin, and 100 U / mL streptomycin. HepG2.2.15 cells were seeded at the same density and used as a medium control. Experimental groups consisted of administration of substances at concentrations specified by the manufacturer. All cultures were grown in a humidified incubator at 37 °C and 5% CO2.

[0175] [Detection of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg)] HepG2 2.2.15 cells were seeded in 12-well microtiter plates. On day 0, cells were washed several times with PBS and treated with tetracycline-free medium containing either the test drug or a solvent control. Each test compound was screened in triplicate at one concentration. On days 3, 6, and / or 9, the medium was removed and replaced with fresh medium containing the compound. After 24 hours, the medium was collected and clarified by centrifugation (Sorvall RT-6000D centrifuge, 1,000 rpm, 5 min). The medium of HepG2 2.2.15 cell cultures was harvested on days 3, 6, and 9 after HBV infection, and the levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in the culture medium were measured using the AXSYM system kit (Abbott Diagnostika).

[0176] [Cell line and culture conditions] - HepAD38 cells In the following experimental examples, when HepAD38 cells (a cell line expressing hepatitis B virus) are used, the cells are maintained in Dulbecco's modified Eagle / F-12 medium (DMEM / F-12, GIBCO BRL / Life Technologies, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS), 100 U / ml penicillin, 100 μg / ml streptomycin, 5 μg / ml ITS, 400 μg / ml G418, and 1.5 g / ml tetracycline. HBV particle production was induced in the HepAD38 cell line by culturing in tetracycline-free medium. When HepAD38 cells were used for HBV collection, tetracycline was removed from the cell culture for 9 days and then treated with the test drug or dimethyl sulfoxide (DMSO) medium control.

[0177] [Detection of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg)] HepAD38 cells were seeded (2x10) in 12-well microtiter plates. 5 Cells / well were seeded and cultured for 3 days in the presence of 0.3 mg tetracycline per ml. On day 0, cells were washed several times with PBS and treated with tetracycline-free medium containing either the test drug or a solvent control. Each test compound was screened in triplicate at one concentration. On days 3, 6, and 9, the medium was removed and replaced with fresh medium containing the compound. After 24 hours, the medium was collected and clarified by centrifugation (Sorvall RT-6000D centrifuge, 1,000 rpm, 5 min). The medium of HepAD38 cell cultures was harvested on days 3, 6, and 9 after HBV infection, and the levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in the culture medium were measured using the AXSYM system kit (Abbott Diagnostika). The presence of HBV DNA in the supernatant medium was examined by RT-PCR.

[0178] [Biomaterials and methods] For in vivo experiments, male C3H / HeN mice were purchased from the National Laboratory Animal Center (Taipei, Taiwan). Animals were maintained under specific pathogen-free conditions at the Laboratory Animal Center, School of Medicine, National Yang Ming Jiao Tong University. All mice were used in accordance with the guidelines for the use of laboratory animals set out by the School of Medicine, National Yang Ming Jiao Tong University. Four mice per cage were fed regular chow and housed in a temperature-controlled room (21 ± 3 °C) with a 12 / 12-h day-night cycle (lights on from 6 am to 6 pm) with a 30-min twilight period.

[0179] [Hydrodynamic injection and serum collection] For in vivo experiments, HBV expression plasmid pAAV / HBV1.2 (genotype A) was used according to Huang et al. (Proc Natl Acad Sci USA 2006;103:17862-17867). Prior to hydrodynamic injection (HDI), all animals were anesthetized using ketamine (0.75 g / kg, Merial) and xylazine (60 μg / kg, Bayer) administered by intramuscular injection. 10 μg of pAAV / HBV1.2 dissolved in PBS at 8% of body weight was injected into the tail vein of mice. Injection time was controlled between 5 and 8 seconds. Approximately 150 μL of serum was collected on the second day and every week after HDI until the end of the experiment. HBsAg, HBeAg, and anti-HBs were measured using serum AXSYM system kits (Abbott Diagnostika). Measurements of anti-HBs were absolute, whereas measurements of HBsAg were relative. The HBsAg positivity threshold was set at a signal-to-noise ratio of 10(1). Statistics were calculated using GraphPad Prism and Microsoft Excel.

[0180] [DNA extraction] Nucleic acids were extracted from cell supernatants using a QIAamp DNA Blood Kit (QIAGEN, 51106, Hilden, Germany). First, 100 μl of cell supernatant was added to a microcentrifuge tube, then 200 μl of AL buffer was added and mixed well by shaking. 20 μl of proteinase K was added and mixed well, the sample was placed in a 56°C incubator for 10 min, the sample was removed and mixed well by shaking, then 200 μl of 100% alcohol was added and mixed well by shaking. The liquid was transferred to a QIAamp spin column and centrifuged at 8000 rpm for 1 min. The liquid was discarded and 500 μl of AW1 buffer was added to wash the QIAamp spin column. Centrifuged at 8000 rpm for 1 min. Centrifuged at maximum speed for 3 min and the remaining liquid was discarded. The QIAamp spin column was transferred to a new microcentrifuge tube, 100 μl of AE buffer was added to the QIAamp spin column membrane, and the mixture was centrifuged at 8000 rpm for 1 minute to obtain an extracted nucleic acid sample.

[0181] Experimental Example 1: In vitro cell line evaluation of entecavir-based compounds

[0182] In Experimental Example 1, five hepatitis treatment groups were prepared and added to HepG2 2.2.15 cells for evaluation. Group 1 was a control group containing only medium (PBS) and no drug compound was added. Group 2 used the conventional drug Entecavir (100 mM) alone. Group 3 (CPD-0148) was targeted with the compound of formula (IA2) (100 mM). Group 4 (CPD-0150) was targeted with the compound of formula (IA3) (100 mM). Group 5 (CPD-0147) was targeted with the compound of formula (IA4) (100 mM). After 6-9 days of culture in the above treatment groups, the levels of Hepatitis B surface antigen (HBsAg) were measured using the AXSYM system kit (Abbott Diagnostika), and the results are shown in Figure 10.

[0183] As shown in FIG. 10, it can be seen that the conventional drug Entecavir (Group 2) can reduce HBsAg levels compared to the control. Furthermore, when the conventional drug Entecavir is modified with the linker and sugar described in the present invention, for example, when the compound of formula (IA2) (CPD-0148; Group 3) or the compound of formula (IA3) (CPD-0150; Group 4) is used, it is found that the HBsAg level is further reduced compared to the unmodified drug Entecavir (Group 2). It should be noted that not all types of linkers and sugars can be used to effectively reduce HBsAg levels. For example, as shown in FIG. 10, when the compound of formula (IA4) (CPD-0147; Group 5) is used, the HBsAg level is similar to the unmodified drug Entecavir (Group 2), and no significant reduction in the HBsAg level was observed. These results suggest that a carbamate bond between the linker and the sugar (>NC(=O)-O-; group 3) or a carbonate bond between the linker and the sugar (-OC(=O)-O-; group 4) may be superior to an ester bond (-RC(=O)-O-; group 5) in further reducing HBsAg levels.

[0184] Experimental Example 2: In vitro cell line evaluation of birinapants

[0185] In Experimental Example 2, eight hepatitis treatment groups were prepared and added to HepG2 2.2.15 cells for evaluation. Group 1 was a control group containing only medium (PBS) and no drug compound was added. Group 2 used the conventional drug birinapant (500 mM) alone. Group 3 (CPD-0004) was subjected to compound of formula (IB1) (500 mM). Group 4 (CPD-0030) was subjected to compound of formula (IB7) (500 mM). Group 5 (CPD-0036) was subjected to compound of formula (IB2) (500 mM). Group 6 (CPD-0133) was subjected to compound of formula (IB3) (500 mM). Group 7 (CPD-0142) was subjected to compound of formula (IB4) (500 mM). Group 8 (CPD-0186) was treated with the compound of formula (IB5) (500 mM). After 6-9 days of culture in the above treatment groups, the levels of Hepatitis B surface antigen (HBsAg) were measured using the AXSYM system kit (Abbott Diagnostika), and the results are shown in Figure 11.

[0186] As shown in FIG. 11, it can be seen that the conventional drug birinapant (group 2) can reduce HBsAg levels compared to the control. Furthermore, when the conventional drug birinapant is modified with the linker and sugar described in the present invention, for example, when using the compound of formula (IB1) (CPD-0004; group 3), the compound of formula (IB2) (CPD-0036; group 5), the compound of formula (IB3) (CPD-0133; group 6), the compound of formula (IB4) (CPD-0142; group 7), and the compound of formula (IB5) (CPD-0186; group 8), it was found that the HBsAg levels were further reduced compared to the unmodified drug birinapant (group 2). It should also be noted that when the monomeric birinapant (formula (IB7); CPD-0030) is modified in the same way as the compound of formula (IB1) (CPD-0004; group 3), a similar reduction in HBsAg levels is not observed. These results suggest that when two glycans (double chains) are attached to a drug via a linker, the reduction in HBsAg levels may be more pronounced.

[0187] Experimental Example 3: In vitro cell line evaluation of tenofovir compounds

[0188] In Experimental Example 3, 13 hepatitis treatment groups were prepared and added to HepG2 2.2.15 cells for evaluation. Group 1 was a control group containing only medium (PBS) and no drug compound was added. Group 2 (Teno-Int) used the conventional drug tenofovir intermediate (100 mM; structure shown in the first step of Figure 3) alone. Group 3 (CPD-0134) was targeted to the compound of formula (IC1) (100 mM). Group 4 (CPD-0136) was targeted to the compound of formula (IC2) (100 mM). Group 5 (CPD-0137) was targeted to the compound of formula (IC3) (510 mM). Group 6 (CPD-0138) was targeted to the compound of formula (IC4) (100 mM). Group 7 (CPD-0139) was targeted to the compound of formula (IC5) (100 mM). Group 8 (CPD-0143) was treated with the compound of formula (IC6) (100 mM). Group 9 (CPD-0144) was treated with the compound of formula (IC7) (100 mM). Group 10 (CPD-0149) was treated with the compound of formula (IC8) (100 mM). Group 11 (CPD-0158) was treated with the compound of formula (IC9) (100 mM). Group 12 (CPD-0169) was treated with the compound of formula (IC10) (100 mM). Group 12 (CPD-0170) was treated with the compound of formula (IC11) (100 mM). After 6-9 days of culture in the above treatment groups, the levels of Hepatitis B surface antigen (HBsAg) were measured using the AXSYM system kit (Abbott Diagnostika), and the results are shown in Figure 12.

[0189] As shown in FIG. 12, it can be seen that the use of tenofovir intermediate (Teno-Int) alone results in a slight reduction in HBsAg levels compared to the control. Furthermore, conventional tenofovir intermediates can be modified with the linkers and sugars described in the present invention to give, for example, compounds of formula (IC1) (CPD-0134; group 3), compounds of formula (IC2) (CPD-0136; group 4), compounds of formula (IC3) (CPD-0137; group 5), compounds of formula (IC4) (CPD-0138; group 6), compounds of formula (IC5) (CPD-0139; group 7), compounds of formula (IC6) (CPD-0143; group 8), compounds of formula (IC7) (CPD-0144; group 9), compounds of formula (IC8) (CPD-0149; group 10), compounds of formula (IC9) (CPD-0158; group 11), compounds of formula (IC10) (CPD-0169; group 12), and compounds of formula (IC11) (CPD-0170; The use of tenofovir intermediate (group 13) leads to a further reduction in HBsAg levels compared to the unmodified tenofovir intermediate (group 2).

[0190] Furthermore, it can be seen that the lowest HBsAg levels were observed when compounds of formula (IC2), formula (IC7), and formula (IC9) were used. These results suggest that the presence of a carbamate bond (>NC(=O)-O-) linking the linker and the glycan and the presence of a triazole group in the linker may be responsible for further reducing the HBsAg levels.

[0191] Experimental Example 4: In vitro cell line evaluation of LCL161-based compounds

[0192] In Experimental Example 4, six hepatitis treatment groups were prepared and added to HepG2 2.2.15 cells for evaluation. Group 1 was a control group containing only medium (PBS) and no drug compound was added. Group 2 used the conventional drug LCL161 (500 mM) alone. Group 3 (CPD-0033) was targeted with compound of formula (ID1) (500 mM). Group 4 (CPD-0130) was targeted with compound of formula (ID2) (500 mM). Group 5 (CPD-0156) was targeted with compound of formula (ID3) (500 mM). Group 6 (CPD-0171) was targeted with compound of formula (ID4) (500 mM). After 6-9 days of culture in the above treatment groups, the levels of Hepatitis B surface antigen (HBsAg) were measured using the AXSYM system kit (Abbott Diagnostika), and the results are shown in Figure 13.

[0193] As shown in Figure 13, it can be seen that the conventional drug LCL161 (group 2) can reduce HBsAg levels compared to the control. Furthermore, when the conventional drug LCL161 is modified with the linker and sugar described in the present invention, for example, when the compound of formula (ID1) (CPD-0033; group 3), the compound of formula (ID2) (CPD-0130; group 4), the compound of formula (ID3) (CPD-0156; group 5), and the compound of formula (ID4) (CPD-0171; group 6) are used, the HBsAg levels are further reduced compared to the unmodified drug LCL161 (group 2). Furthermore, it can be seen that the compounds of formula (ID3) and formula (ID4) have the lowest HBsAg levels. These results suggest that the presence of a carbamate bond (>NC(=O)-O-) linking the linker and the glycan, the presence of a triazole group in the linker, and control of the number of sugars and the number of glycans may further reduce HBsAg levels.

[0194] Experimental Example 5: In vitro cell line evaluation of telbivudine and adefovir compounds

[0195] In Experimental Example 5, five hepatitis treatment groups were prepared and added to HepG2 2.2.15 cells for evaluation. Group 1 was a control group containing only medium (PBS) and no drug compound was added. Group 2 used the conventional drug telbivudine (200 mM) alone. Group 3 (CPD-0117) was targeted with the compound of formula (IE1) (200 mM). Group 4 (Ade-Int) used the intermediate of the conventional drug adefovir (100 mM; structure shown in the first step of Figure 6) alone. Group 4 (CPD-0146) was targeted with the compound of formula (IF1) (200 mM). After 6-9 days of culture in the above treatment groups, the levels of hepatitis B surface antigen (HBsAg) were measured using the AXSYM system kit (Abbott Diagnostika), and the results are shown in Figure 14.

[0196] As shown in Figure 14, it can be seen that further modification of the conventional drug telbivudine (group 2) with a linker and sugar as described in the present invention can further reduce HBsAg levels, for example when using a compound of formula (IE1) (CPD-0117; group 3). Similarly, further modification of the conventional adefovir intermediate (group 3) with a linker and sugar as described in the present invention can further reduce HBsAg levels, for example when using a compound of formula (IF1) (CPD-0146; group 4).

[0197] Experimental Example 6: In vitro cell line evaluation of drug conjugate combinations

[0198] In Experimental Example 6, three hepatitis treatment groups were prepared and added to HepG2 2.2.15 cells for evaluation. Group 1 was a control group containing only medium (PBS) and no drug compound was added. Group 2 used the conventional drug Entecavir (100 mM) alone. Group 3 (CPD-0005+CPD-0004) used a combination of the compound of formula (IA1) (100 mM) and the compound of formula (IB1) (100 mM). After 6-9 days of culture in the above treatment groups, the levels of Hepatitis B surface antigen (HBsAg) and Hepatitis B e antigen (HBeAg) were measured using the AXSYM system kit (Abbott Diagnostika), and the results are shown in Figure 15.

[0199] As shown in Figure 15, it can be seen that the conventional drug Entecavir is effective in reducing HBsAg and HBeAg levels compared to the control. However, when the drug conjugate of formula (IA1) is used in combination with the drug conjugate of formula (IB1), it can be seen that the HBsAg and HBeAg levels are further reduced. These results suggest that the drug conjugate of the present invention is more effective in reducing HBsAg and HBeAg levels when used in combination than the conventional drug.

[0200] Experimental Example 7: In vitro cell line evaluation of drug conjugate combinations

[0201] In Experimental Example 7, three hepatitis treatment groups were prepared and added to HepAD38 cells for evaluation. Group 1 was a control group containing only medium (PBS) and no drug compound was added. Group 2 was treated with a combination of conventional drugs Entecavir (25 nM) and Birinapant (25 nM). Group 3 (CPD-0005+CPD-0004) was treated with a combination of compound of formula (IA1) (25 nM) and compound of formula (IB1) (25 nM). After 6-9 days of culture in the above treatment groups, the levels of Hepatitis B surface antigen (HBsAg), Hepatitis B e antigen (HBeAg) were measured using the AXSYM system kit (Abbott Diagnostika), and the presence of HBV DNA in the supernatant medium was examined by RT-PCR. The results are shown in Figure 16A and Figure 16B.

[0202] As shown in FIG. 16A, it can be seen that the combination of conventional drugs, entecavir and birinapant, is effective in lowering HBsAg and HBeAg levels compared to the control. However, it can be seen that the combination of the drug conjugate of formula (IA1) and the drug conjugate of formula (IB1) further reduces HBsAg and HBeAg levels. These results suggest that the combination of the drug conjugate of the present invention is more effective in lowering HBsAg and HBeAg levels than the conventional drugs. Furthermore, as shown in FIG. 16B, it can be seen that the combination of the drug conjugate of formula (IA1) and formula (IB1) of the present invention is more effective in lowering HBV DNA levels than the combination of conventional drugs, entecavir and birinapant.

[0203] Experimental Example 8: Biological evaluation of combined drug conjugates

[0204] In Experimental Example 8, three hepatitis treatment groups were prepared and administered to C3H mice (intraperitoneal (ip) administration). Group 1 was a control group without drug compounds (PBS as control). Group 2 was administered a combination of a conventional drug, Entecavir (1.5 mg / kg, ip), and a conventional drug, Birinapant (10 mg / kg, ip). Group 3 was administered a combination of a drug conjugate of formula (IA1) (3.8 mg / kg, ip) and a drug conjugate of formula (IB1) of the present invention (16.6 mg / kg, ip) (CPD-0005+CPD-0004). HBsAg, HBeAg and HBV DNA levels of mice were measured on day 2 (after HDI), day 14, day 21, day 63, day 77, day 105, day 133 and day 151. The results are shown in Figures 17A to 19B. Additionally, hair loss in the C3H mice was also assessed, and the hair loss observations are shown in Figure 20. Anti-HBsAg levels in the mice were also measured on day 151, and the results are shown in Figure 21.

[0205] As shown in Figures 17A, 17B, 18A and 18B, when the drug conjugate of formula (IA1) and the drug conjugate of formula (IB1) are used in combination, both HBsAg and HBeAg levels are further reduced compared to the conventional drugs (combination of entecavir and birinapant). The reduction in HBsAg and HBeAg levels is more significant after day 63. Furthermore, as shown in Figures 19A and 19B, when the drug conjugate of formula (IA1) of the present invention is used in combination with the drug conjugate of formula (IB1), the effect of reducing HBV DNA levels is significantly superior to the conventional drugs entecavir and birinapant. These results demonstrate that a synergistic effect can be obtained when the drug conjugate of the present invention is used in combination in animals (in vivo), and that the combination of the drug conjugates is more effective in reducing HBsAg and HBeAg levels than the conventional drugs. This result is consistent with the results obtained in the in vitro cell line study.

[0206] Furthermore, as shown in Figure 20, it can be seen that the conventional drugs Entecavir and Birinapant are effective in reducing HBeAg and HBV DNA levels, but significant hair loss was also observed in C3H mice. In comparison, the drug conjugates of formula (IA1) and (IA2) of the present invention did not show hair loss in C3H mice even after 95 days. These results suggest that the drug conjugates of the present invention are effective in reducing HBsAg, HBeAg and HBV DNA levels and have few side effects.

[0207] Furthermore, as shown in Figure 21, even when conventional drugs, entecavir and birinapant, were used in combination, the anti-HBsAg level obtained was comparable to that of the control. In contrast, when the drug conjugate of formula (IA1) and the drug conjugate of formula (IB1) were used in combination (CPD-0005+CPD-0004), a significantly higher anti-HBsAg level was observed. These results suggest that the composition containing the drug conjugate of formula (IA1) and formula (IB1) is effective in inducing an immune response against the hepatitis B virus, whereas such an effect is not observed with conventional drugs.

[0208] Experimental Example 9: In vivo evaluation of the compound of formula (IA1)

[0209] In Experiment 9, three hepatitis treatment groups were prepared and administered to C3H mice (intraperitoneal (ip) administration). Group 1 was a control group without drug compound (PBS as control). Group 2 used the conventional drug Entecavir (1.5 mg / kg, ip) alone. Group 3 used the drug conjugate of formula (IA1) (3.8 mg / kg, ip). HBsAg, HBeAg and HBV DNA levels of mice were measured on days 2 (post HDI), 14, 21, 28, 35, 42, 49 and 56. The results are shown in Figures 22A to 24B.

[0210] As shown in Figures 22A, 22B, 23A, and 23B, it can be seen that when the conventional drug Entecavir is modified with the drug conjugate (CPD-0005) of formula (IA1) of the present invention, HBsAg and HBeAg levels are further reduced compared to the conventional drug. The reduction in HBsAg and HBeAg levels is more significant after day 49. Furthermore, as shown in Figures 24A and 24B, it was also found that the drug conjugate of formula (IA1) of the present invention appears to be more effective in reducing HBV DNA levels compared to the conventional drug Entecavir. These results demonstrated that the drug conjugate of the present invention is more effective in reducing HBsAg, HBeAg, and HBV DNA levels than the conventional drug when used in animals (in vivo). The results are consistent with those obtained in the in vitro cell line study.

[0211] Experimental Example 10: In vivo evaluation of the compound of formula (IB1)

[0212] In Experimental Example 10, three hepatitis treatment groups were prepared and administered to C3H mice (intraperitoneal (ip) administration). Group 1 was a control group without drug compound (PBS as control). Group 2 used the conventional drug birinapant (10 mg / kg, ip) alone. Group 3 used the drug conjugate of formula (IB1) (16.6 mg / kg, ip). HBsAg, HBeAg and HBV DNA levels of mice were measured on days 2 (post HDI), 14, 21, 28, 35, 56, 63 and 70. The results are shown in Figures 25A to 27B.

[0213] As shown in Figures 25A, 25B, 26A, and 26B, it can be seen that when the conventional drug birinapant is modified to form the drug conjugate (CPD-0004) of formula (IB1) of the present invention, HBsAg and HBeAg levels are further reduced compared to the conventional drug. The reduction in HBsAg and HBeAg levels is more significant after day 56. Furthermore, as shown in Figures 27A and 27B, it was also found that the drug conjugate of formula (IB1) of the present invention appears to be more effective in reducing HBV DNA levels compared to the conventional drug birinapant. These results demonstrated that the drug conjugate of the present invention is more effective in reducing HBsAg, HBeAg, and HBV DNA levels than the conventional drug when used in animals (in vivo). The results are consistent with those obtained in the in vitro cell line study.

[0214] According to exemplary embodiments of the present invention, the drug conjugate or pharmaceutical composition of the present invention is more effective than conventional drugs in treating hepatitis, such as hepatitis B. The drug conjugate or pharmaceutical composition of the present invention can further reduce hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) levels, and HBV DNA levels, while improving hepatitis B surface antibody (anti-HBsAg) levels, compared to conventional hepatitis B treatments. Furthermore, when the first drug conjugate (including the HBV drug) is combined with the second drug conjugate (including the IAP antagonist) in a pharmaceutical composition, at least the same or lower HBsAg, HBeAg, and HBV DNA levels can be achieved, while reducing side effects (hair loss). In some embodiments, when either the first drug conjugate (including the HBV drug) or the second drug conjugate (including the IAP antagonist) is combined with a conventional drug (non-denatured drug compound), HBsAg, HBeAg, and HBV DNA levels can also be further reduced, compared to the conventional drug alone.

[0215] It will be apparent to one skilled in the art that various modifications and variations can be made to the disclosed embodiments without departing from the scope or spirit of the present invention. In view of the foregoing, it is intended that the present invention cover modifications and variations provided they come within the scope of the following claims and their equivalents. [Industrial Applicability]

[0216] The drug conjugate or pharmaceutical composition of the present invention can be applied to the treatment of hepatitis.

Claims

1. A drug conjugate having the structure shown in formula (I): Z-(linker-[R] ｍ ) ｎ Formula (I) wherein in formula (I), Z is a drug compound, R is a sugar, and m and n are independently integers from 1 to 6.

2. The drug compound Z is a hepatitis virus targeted drug, a hepatitis B virus (HBV) drug, a hepatitis C virus (HCV) drug, an inhibitor of apoptosis protein (IAP) antagonist, a multidrug resistance (MDR) inhibitor, or an analog, precursor, prodrug, or derivative thereof; The drug conjugate of claim 1.

3. The drug compound Z is selected from the group consisting of tenofovir, tenofovir diisoproxil, tenofovir alafenamide, entecavir, telbivudine, adefovir, adefovir dipivoxil, lamivudine, interferon-α-2A, interferon-α-2B, birinapant, monomeric birinapant, zebinapant, LCL161, GDC-0152, GDC-0917, CUDC-427, APG-1387, DeBio-1143, sergantolimod, BI-82, and zosquivir. darupervir, glecaprevir, pibrentasvir, ombitasvir, paritaprevir, ritonavir, sovosbuvir, velapatasvir, ledipasvir, voxilaprevir, daclatasvir, asunaprevir, telaprevir, elbasvir, grazoprevir, ribavarin, telaprevir, EDP-239, alisporivir, filibirubir, melicitabine, danoprevir, velpatasvir, or analogs, precursors, prodrugs, or derivatives thereof; The drug conjugate of claim 1.

4. The sugar R is selected from the group consisting of monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides, polysaccharides, or derivatives thereof; The drug conjugate of claim 1.

5. The linker may be substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, aryl alkyl, aryl alkenyl, aryl alkynyl, heteroaryl alkyl, heteroaryl alkenyl, heteroaryl alkynyl, heterocyclyl alkyl, heterocyclyl alkenyl, heterocyclyl alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylaryl alkyl, alkylaryl alkenyl, alkylaryl alkynyl, alkenylaryl alkyl, alkenylaryl alkynyl, alkynylaryl alkyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkylheteroaryl alkyl, alkylheteroaryl alkenyl, alkylheteroaryl alkynyl. wherein one or more methylenes are selected from the group consisting of O, S, S(O), SO, S(O), ... ２ , N(R ８ ), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, ８ is hydrogen, acyl, aliphatic or substituted aliphatic; The drug conjugate of claim 1.

6. The linker of formula (I) is L １ (L ２ ) ｐ group, wherein L １ The group is attached to the drug compound Z, and L ２ The group is L １ R is an integer from 1 to 4; １ The group may comprise a carbamate, amide, ester, amidine, carboximidate, sulfonate, sulfate, sulfonamide, thioether, ether, imine, imide, thioamide, carbonate, thiocarbamide, carbamide, triazole, amine, oxime, thiocarbonate, thioester, or thiocarbamate group; L ２ The group is selected from the group consisting of a single bond, an alkyl having 1 to 50 carbons, an alkoxy having 1 to 50 carbons, an alkenyl having 2 to 50 carbons, and an alkenyloxy having 2 to 50 carbons, and in the above groups, the alkyl, alkoxy, alkenyl, and alkenyloxy may be linear or branched, and may have at least one -CH ２ - may be substituted by -CO-, -COO-, -OCO-, -S-, -O- or -NH-, and at least one -CH ２ CH ２ - may be replaced by -CH=CH- or -C≡C-, and the above-mentioned -CH ２ At least one hydrogen in -, -CH= or -NH- may be substituted with halogen, trifluoromethyl, alkyl having 1 to 20 carbons, alkoxy having 1 to 20 carbons, or alkanoyl having 1 to 20 carbons, and at least one -O- may be substituted with a phosphate, phosphonate, phosphonate, phosphinate, phosphine oxide, phosphoamide, phosphoramidate, phosphite, phosphonite, phosphine, aminophosphine, phosphoramidite, phosphonamidite, phosphonamide, phosphinamide, or phosphorodiamidite group; The drug conjugate of claim 1.

7. The linker is any one selected from the group consisting of formula (LX1) to formula (LX30): The drug conjugate of claim 1. 【Chemistry 1】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 In formulae (LX1) to (LX30), Z is the position where the linker is attached to the drug, and R is the position where the linker is attached to the sugar.

8. the sugar R is selected from the group consisting of monosaccharides, disaccharides, and trisaccharides, wherein the monosaccharides, disaccharides, and trisaccharides are independently unsubstituted or substituted with an acetyl group or an N-acetyl group; wherein the drug Z is selected from the group consisting of entecavir, adefovir, telbivudine, lamivudine, tenofovir, GDC-0152, DeBio-1143, LCL161, birinapant, or analogs, precursors, prodrugs, or derivatives thereof; The drug conjugate of claim 7.

9. In the drug conjugate, the drug compound Z is bonded to the linker to form a carbamate bond (>N-C(=O)-O-), an amide bond (R-C(=O)-N<), a carbonate bond (RO-C(=O)-OR'), or an ester bond (R-C(=O)-OR'). The drug conjugate of claim 1.

10. In the drug conjugate, the drug compound Z is bonded to the linker to form a carbamate bond (>N-C(=O)-O-) or an ester bond (R-C(=O)-OR'). The drug conjugate of claim 9.

11. The linker comprises at least a triazole. The drug conjugate of claim 1.

12. The structure shown in formula (I) may be further represented by formula (II): The drug conjugate of claim 1. Z-(linker-[R] ｍ ) ２ Formula (II) wherein in formula (II), Z is a drug compound, R is a sugar, and m is independently an integer from 1 to 3.

13. The structure represented by formula (I) may be further represented by any one of formulas (IA1) to (IA4): The drug conjugate of claim 1. 【Chemistry 2】

14. The structure represented by formula (I) may be further represented by any one of formulas (IB1) to (IB11): The drug conjugate of claim 1. 【Chemistry 3】 【change】 【change】 【change】 【change】

15. The structure shown in formula (I) may be further represented by any one of formulas (IC1) to (IC11): The drug conjugate of claim 1. 【Chemistry 4】 【change】 【change】 【change】

16. The structure shown in formula (I) may be further represented by any one of formulas (ID1) to (ID5): The drug conjugate of claim 1. 【Chemistry 5】 【change】

17. The structure represented by formula (I) may be further represented by formula (IE1): The drug conjugate of claim 1. 【Chemistry 6】

18. The structure represented by formula (I) may be further represented by formula (IF1) or formula (IF2): The drug conjugate of claim 1. 【Chemistry 7】

19. The structure represented by formula (I) may be further represented by formula (IG1): The drug conjugate of claim 1. 【Chemistry 8】

20. The structure represented by formula (I) may be further represented by any one of formulas (IH1) to (IH3): The drug conjugate of claim 1. 【Chemistry 9】

21. The structure represented by formula (I) may be further represented by formula (IJ1) or formula (IJ2): The drug conjugate of claim 1. 【Chemistry 10】

22. A pharmaceutical composition comprising an active ingredient, said active ingredient comprising a drug conjugate having the structure shown in formula (I). Z-(linker-[R] ｍ ) ｎ Formula (I) In the formula (I), Z is a drug compound, R is a sugar, and m and n are independently integers from 1 to 6; At least one or more pharma- ceutically acceptable excipients.

23. The drug conjugate comprises a first drug conjugate represented by formula (I): In formula (I), drug compound Z is a first drug compound X selected from the group consisting of tenofovir, tenofovir diisoproxil, tenofovir alafenamide, entecavir, telbivudine, adefovir, adefovir dipivoxil, lamivudine, interferon-α-2A, interferon-α-2B, sergantolimod, BI-82 and zosquidar, or analogs, precursors, prodrugs, derivatives thereof; 23. The pharmaceutical composition of claim 22.

24. The first drug conjugate represented by formula (I) may further be represented by any one of formulas (IA1) to (IA4), any one of formulas (IC1) to (IC11), or formula (IF1), or formula (IF2), or formula (IG1), 24. The pharmaceutical composition of claim 23. 【Chemistry 11】 【change】 【change】 【change】 【change】

25. The drug conjugate further comprises a second drug conjugate represented by formula (I), wherein the drug compound Z is a second drug compound Y selected from the group consisting of birinapant, monomeric birinapant, zebinapant, LCL161, GDC-0152, GDC-0917, CUDC-427, APG-1387, DeBio-1143, or analogs, precursors, prodrugs, or derivatives thereof; 25. The pharmaceutical composition of claim 24.

26. The second drug conjugate represented by formula (I) may further be represented by any one of formulas (IB1) to (IB11), or any one of formulas (ID1) to (ID5), or any one of formulas (IH1) to (IH3), or formula (IJ1) or formula (IJ2):

26. The pharmaceutical composition of claim 25. 【Chemistry 12】 【change】 【change】 【change】 【change】 【change】 【change】 【change】

27. The first drug conjugate is a compound represented by formula (IA1), and the second drug conjugate is a compound represented by formula (IB1).

27. The pharmaceutical composition of claim 26. 【Chemistry 13】

28. the first drug conjugate and the second drug conjugate are each administered at a dose of 0.1 mg / kg to 100 mg / kg; 28. The pharmaceutical composition of claim 27.

29. The sugar R is a monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, polysaccharide, or derivative thereof; 23. The pharmaceutical composition of claim 22.

30. The linker may be substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, aryl alkyl, aryl alkenyl, aryl alkynyl, heteroaryl alkyl, heteroaryl alkenyl, heteroaryl alkynyl, heterocyclyl alkyl, heterocyclyl alkenyl, heterocyclyl alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylaryl alkyl, alkylaryl alkenyl, alkylaryl alkynyl, alkenylaryl alkyl, alkenylaryl alkynyl, alkynylaryl alkyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkynylaryl alkenyl, alkylheteroaryl alkyl, alkylheteroaryl alkenyl, alkylheteroaryl alkynyl. wherein one or more methylenes are selected from the group consisting of O, S, S(O), SO, S(O), ... ２ , N(R ８ ), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, ８ is hydrogen, acyl, aliphatic or substituted aliphatic; 23. The pharmaceutical composition of claim 22.

31. The pharmaceutical composition is prepared into tablets, capsules, granules, powders, solutions, syrups, sprays, injections or inhalants.

23. The pharmaceutical composition of claim 22.

32. The at least one or more pharma- ceutically acceptable excipients are selected from the group consisting of fillers, bulking agents, binders, mixing agents, surfactants, emulsifiers, dispersing agents, antifoaming agents, lubricants, non-adherents, blending agents, coating materials, glidants, anti-adherents, diluents, dyes, pigments, dispersants, wetting agents, and combinations thereof; 23. The pharmaceutical composition of claim 22.

33. 13. Use of the drug conjugate of claim 1 for the manufacture of a medicament for the treatment of hepatitis.

34. Use of the pharmaceutical composition according to claim 22 for the manufacture of a medicament for the treatment of hepatitis.

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