Anthracycline derivative, and preparation method therefor and use thereof

Abstract

Provided in the present application are a new cytotoxic compound, a preparation method therefor and the use thereof. Specifically, provided in the present application are an anthracycline derivative as represented by general formula (I), and a preparation method therefor, and the use thereof in the preparation of a drug for treating and / or preventing tumors.

Description

Anthracycline derivatives, their preparation methods and applications

[0001] This application claims priority to Chinese Patent Application No. 2024110373609, filed July 30, 2024; Chinese Patent Application No. 202411338856X, filed September 24, 2024; Chinese Patent Application No. 2025100973019, filed January 21, 2025; and Chinese Patent Application No. 2025110081696, filed July 21, 2025. The full text of the aforementioned Chinese patent applications is incorporated herein by reference. Technical Field

[0002] This application belongs to the pharmaceutical field, specifically relating to an anthracycline derivative, its preparation method, and its uses. Background Technology

[0003] Anthracyclines are a class of antitumor drugs extracted from or semi-synthesized by Streptomyces bacteria. They are a large class of chemotherapy drugs used clinically and are widely applied in cancer treatment. Currently marketed anthracycline drugs include doxorubicin, daunorubicin, epirubicin, idarubicin, arubicin, pentorubicin, pirarubicin, and amrubicin. Doxorubicin, as a classic broad-spectrum antitumor drug, has been used clinically for over 70 years and is often combined with other chemotherapy drugs to treat acute lymphoblastic leukemia, acute myeloid leukemia, Hodgkin's and non-Hodgkin's lymphoma, breast cancer, lung cancer, ovarian cancer, soft tissue sarcoma, osteosarcoma, rhabdomyosarcoma, nephroblastoma, neuroblastoma, bladder tumor, thyroid tumor, choriocarcinoma, prostate cancer, testicular cancer, gastric cancer, and liver cancer.

[0004] The molecular skeleton of anthraquinone compounds consists of a four-membered ring based on anthraquinone structure linked to a six-membered sugar ring via a glycosidic bond. Their primary mechanism of action is as inhibitors of topoisomerase II, inducing DNA double-strand breaks or chromatin damage, thereby leading to apoptosis. Specifically, when anthraquinone molecules are taken up into cells, the four-membered ring structure intercalates between DNA base pairs, while the sugar units bind to the minor groove of the DNA double helix and interact with topoisomerase II, inhibiting DNA replication and transcription. On the other hand, anthraquinone molecules attaching to the DNA surface can cause histone detachment, disrupting chromatin structure and thus affecting gene regulation.

[0005] Although anthracyclines have a wide range of cancer indications, they also face many challenges in clinical application, such as cardiotoxicity. Furthermore, the activity of anthracycline molecules currently used clinically is generally low, which limits their application in drug formulations that require high activity of the active ingredient, such as antibody-drug conjugates.

[0006] Therefore, it is necessary and has broad application prospects to develop anthracycline molecules with moderate tumor cell killing activity and low or no cardiotoxicity. Summary of the Invention

[0007] The first aspect of this application provides a compound of formula (I), a pharmaceutically acceptable salt thereof, and an isomer thereof:

[0008] in,

[0009] R1 is selected from H and C. 1-6 Alkoxy;

[0010] R2 is selected from O and NH;

[0011] R3 is selected from C 1-6 Alkyl, hydroxy C 1-6 Alkyl, -C(O)C 1-6 Alkyl, -C(O)NR n1 R n2 , wherein, the C 1-6 Alkyl groups may be optionally replaced by hydroxyl groups or -OC(O)CH2NH2;

[0012] R n1 and R n2 Each is independently selected from H and C. 1-6 Alkyl, 3- to 8-membered cycloalkyl; wherein, the C 1-6 Alkyl groups, 3 to 8-membered cycloalkyl groups optionally coated with hydroxyl groups, hydroxyl C 1-6 The alkyl group is substituted with one or more substituents selected from alkyl, amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, and 5- to 10-membered heteroaryl, wherein the amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, and 5- to 10-membered heteroaryl are optionally selected from C10. 1-6 Alkyl, amino, hydroxy C 1-6 One or more substituents in the alkyl group are substituted; or,

[0013] R n1 and R n2 The atoms bonded to it together form a 3- to 8-membered heterocyclic group, wherein the 3- to 8-membered heterocyclic group is optionally bonded to C 1-6 Alkyl substitution, wherein the C 1-6 Alkyl groups may be optionally substituted with amino or hydroxyl groups;

[0014] R4 and R5 are each independently selected from H and C. 1-6 Alkyl, 3- to 8-membered cycloalkyl, 3- to 8-membered heterocyclic, wherein the C 1-6The alkyl group is optionally substituted with a 6- to 10-membered aryl group, a 4- to 10-membered bridged cycloyl group, a 3- to 8-membered cycloalkyl group, or a 5- to 10-membered heteroaryl group; wherein the 6- to 10-membered aryl group, the 4- to 10-membered bridged cycloyl group, or the 5- to 10-membered heteroaryl group is optionally selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 The substance is substituted by one or more of the following substituents: 3 to 8-membered heterocyclic group, halogen, cyano, 3 to 8-membered cycloalkyl, -NO2, and amino.

[0015] Alternatively, R4 and R5, together with the nitrogen atom to which they are attached, form a 3- to 8-membered heterocyclic group or a 3- to 8-membered heteroaryl group, wherein the 3- to 8-membered heterocyclic group or the 3- to 8-membered heteroaryl group is optionally bonded by CN or C. 1-6 Alkoxy, C 1-6 Alkyl, -C(O)OC 1-6 Alkyl, -C(O)C 3-8 Cycloalkyl, -C(O)NR n3 R n4 -C(O)C 1-6 One or more substituents in the alkyl group are replaced, wherein the C 1-6 Alkyl groups are optionally surrounded by 3 to 8-membered heterocyclic groups, C 1-6 Alkoxy, NR n3 R n4 It is substituted by one or more substituents among 5- to 10-membered heteroaryl, amino, and hydroxyl groups;

[0016] R n3 and R n4 Each is independently selected from H and C. 1-6 Alkyl, -C(O)C 1-6 alkyl;

[0017] Furthermore, the compound of formula I is not any of the following compounds:

[0018] This application provides a compound of formula (I), a pharmaceutically acceptable salt thereof, and an isomer thereof:

[0019] in,

[0020] R1 is selected from H and C. 1-6 Alkoxy;

[0021] R2 is selected from O and NH;

[0022] R3 is selected from C 1-6 Alkyl, hydroxy C 1-6 Alkyl, -C(O)C 1-6 Alkyl, -C(O)NR n1 Rn2 Wherein C 1-6 Alkyl groups may be optionally replaced by hydroxyl groups or -OC(O)CH2NH2;

[0023] R n1 and R n2 Each is independently selected from H and C. 1-6 Alkyl, 3- to 8-membered cycloalkyl, 6- to 10-membered aryl; wherein, the C 1-6 Alkyl, 3- to 8-membered cycloalkyl, 6- to 10-membered aryl, optionally with hydroxyl, hydroxyl C 1-6 The alkyl group is substituted with one or more substituents selected from alkyl, amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, 5- to 10-membered heteroaryl, and 6- to 10-membered aryl, wherein the amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, 5- to 10-membered heteroaryl, and 6- to 10-membered aryl are optionally selected from C10. 1-6 Alkyl, amino, hydroxy C 1-6 One or more substituents in the alkyl group are substituted;

[0024] Or, R n1 and R n2 The atoms bonded to it together form a 3- to 8-membered heterocyclic group, wherein the 3- to 8-membered heterocyclic group is optionally bonded to C 1-6 Alkyl substitution, wherein the C 1-6 Alkyl groups may be optionally substituted with amino or hydroxyl groups;

[0025] R4 and R5 are each independently selected from H and C. 1-6 Alkyl, -C(O)C 1-6 Alkyl, 3- to 8-membered cycloalkyl, 3- to 8-membered heterocyclic, wherein the C 1-6 The alkyl group is optionally substituted with a 6- to 10-membered aryl group, a 4- to 10-membered bridged cycloyl group, a 3- to 8-membered cycloalkyl group, a 5- to 10-membered heteroaryl group, and a 3- to 8-membered heterocyclic group; wherein the 6- to 10-membered aryl group, the 4- to 10-membered bridged cycloyl group, and the 5- to 10-membered heteroaryl group are optionally selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, halogenated C 1-6 The substance is substituted by one or more substituents selected from alkyl, cyano, 3- to 8-membered cycloalkyl, -NO2, and amino groups;

[0026] Alternatively, R4 and R5, together with the nitrogen atom to which they are attached, form a 3- to 8-membered heterocyclic group or a 3- to 8-membered heteroaryl group, wherein the 3- to 8-membered heterocyclic group or the 3- to 8-membered heteroaryl group is optionally bonded by CN or C. 1-6 Alkoxy, C 1-6Alkyl, -C(O)OC 1-6 Alkyl, -C(O)C 3-8 Cycloalkyl, -C(O)NR n3 R n4 -C(O)C 1-6 One or more substituents in the alkyl group are replaced, wherein the C 1-6 Alkyl groups are optionally surrounded by 3 to 8-membered heterocyclic groups, C 1-6 Alkoxy, NR n3 R n4 It is substituted by one or more substituents among 5- to 10-membered heteroaryl, amino, and hydroxyl groups;

[0027] R n3 and R n4 Each is independently selected from H and C. 1-6 Alkyl, -C(O)C 1-6 alkyl;

[0028] Furthermore, the compound of formula I is not any of the following compounds:

[0029] In certain embodiments of this application, the compound represented by formula (I), its pharmaceutically acceptable salt, its isomer, is selected from formula (IA):

[0030] Wherein R1, R2, R3, R4, and R5 are as defined above, and the compound of formula (IA) is not any of the following compounds:

[0031] In some embodiments of this application, R1 is -OCH3.

[0032] In some embodiments of this application, R1 is -OCH3 or H.

[0033] In some embodiments of this application, R2 is 0.

[0034] In some embodiments of this application, R3 is selected from -C(O)C 1-6 Alkyl and -C(O)NR n1 R n2 Wherein C 1-6 The alkyl group may optionally be replaced by a hydroxyl group and -OC(O)CH2NH2; the C 1-6 Alkyl groups are preferably replaced by hydroxyl groups.

[0035] In some embodiments of this application, R n1 H stands for H independently.

[0036] In some embodiments of this application, Rn2 Selected independently from C 1-6 Alkyl and 6-10 aryl groups; the C 1-6 The alkyl and 6-10 aryl groups are optionally substituted with one or more substituents selected from hydroxyl, amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, 5- to 10-membered heteroaryl, and 6-10 aryl groups; wherein the amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, and 6-10 aryl groups are optionally selected from C 1-6 The alkyl and amino groups are substituted with one or more substituents; preferably, the C 1-6 The alkyl and 6-10 aryl groups are optionally substituted with one or more substituents selected from hydroxyl, amino, and 6-10 aryl groups; wherein the amino and 6-10 aryl groups are optionally selected from C 1-6 It is substituted by one or more substituents in alkyl and amino groups.

[0037] In some embodiments of this application, R n1 and R n2 The atoms bonded to it together form a 3- to 8-membered heterocyclic group, wherein the 3- to 8-membered heterocyclic group is optionally bonded to C 1-6 Alkyl substitution, wherein the C 1-6 Alkyl groups may be optionally substituted with amino and hydroxyl groups.

[0038] In some embodiments of this application, R3 is selected from...

[0039] In some embodiments of this application, R3 is selected from...

[0040] In some embodiments of this application, R3 is selected from...

[0041] In some embodiments of this application, R3 is selected from...

[0042] In some embodiments of this application, R4 is independently selected from C. 1-6 Alkyl and -C(O)C 1-6 Alkyl; preferably C 1-6 Alkyl group, more preferably CH3.

[0043] In some embodiments of this application, R5 is independently selected from C. 1-6 Alkyl, 3- to 8-membered cycloalkyl and 3- to 8-membered heterocyclic groups;

[0044] Wherein, the C 1-6The alkyl group is optionally substituted with a 6- to 10-membered aryl group, a 4- to 10-membered bridged cycloyl group, a 3- to 8-membered cycloalkyl group, a 5- to 10-membered heteroaryl group, and a 3- to 8-membered heterocyclic group; said 6- to 10-membered aryl group, 4- to 10-membered bridged cycloyl group, and 5- to 10-membered heteroaryl group are optionally selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, halogenated C 1-6 Substituted with one or more of the following substituents: alkyl, cyano, 3- to 8-membered cycloalkyl, -NO2, and amino;

[0045] R n3 and R n4 Each is independently selected from H and C. 1-6 Alkyl and -C(O)C 1-6 alkyl;

[0046] Preferably, R5 is independently selected from C 1-6 Alkyl groups and 3 to 8-membered heterocyclic groups, wherein the C 1-6 The alkyl group is optionally substituted with a 6- to 10-membered aryl group, a 4- to 10-membered bridged cycloyl group, a 5- to 10-membered heteroaryl group, and a 3- to 8-membered heterocyclic group; wherein the 6- to 10-membered aryl group, the 4- to 10-membered bridged cycloyl group, and the 5- to 10-membered heteroaryl group are optionally selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, halogenated C 1-6 Substituted with one or more of the following substituents: alkyl, cyano, 3- to 8-membered cycloalkyl, -NO2, and amino;

[0047] R n3 and R n4 Each is independently selected from H and C. 1-6 alkyl;

[0048] More preferably, R5 is independently C 1-6 Alkyl; wherein, the C 1-6 The alkyl group is optionally substituted with a 6- to 10-membered aryl group and a 5- to 10-membered heteroaryl group; wherein the 6- to 10-membered aryl group and the 5- to 10-membered heteroaryl group are optionally selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, halogenated C 1-6Substituted with one or more of the following substituents: alkyl, cyano, 3- to 8-membered cycloalkyl, -NO2, and amino;

[0049] R n3 and R n4 Each is independently selected from H and C. 1-6 alkyl;

[0050] For example, R5 is independently C 1-6 Alkyl; wherein, the C 1-6 The alkyl group may optionally be substituted with phenyl, pyrimidinyl, or pyridinyl groups; wherein the phenyl, pyrimidinyl, or pyridinyl groups are C-substituted. 1-6 Alkoxy, -NR n3 R n4 -CH2-NR n3 R n4 Substitution with 3 to 8-membered heterocyclic groups, halogens, cyano groups, 3 to 8-membered cycloalkyl groups, and -NO2.

[0051] In some embodiments of this application, R4, R5, together with the nitrogen atom to which they are attached, form a 3- to 8-membered heterocyclic group;

[0052] The 3- to 8-membered heterocyclic groups are optionally replaced by CN, C 1-6 Alkyl, -C(O)C 3-8 Cycloalkyl, -C(O)NR n3 R n4 -C(O)C 1-6 One or more substituents in the alkyl group are substituted; the C 1-6 Alkyl groups are optionally C 1-6 It is substituted by one or more substituents among alkoxy, amino, and hydroxyl groups; R n3 and R n4 Each is independently selected from H and C. 1-6 alkyl;

[0053] Preferably, the 3- to 8-membered heterocyclic group is optionally replaced by CN, -C(O)C 3-8 Cycloalkyl, -C(O)C 1-6 It is replaced by one or more substituents in the alkyl group.

[0054] In some embodiments of this application, -NR4R5 has the following structure:

[0055] In some embodiments of this application, -NR4R5 is selected from the following structures:

[0056] In some embodiments of this application, -NR4R5 has the following structure:

[0057] On the other hand, this application provides a compound represented by formula (II), a pharmaceutically acceptable salt thereof, and an isomer thereof:

[0058] in,

[0059] R4 is selected from H and C. 1-6 Alkyl group, preferably C 1-6 Alkyl groups, more preferably CH3;

[0060] Ring A is selected from C6-C 10 Aryl and 5 to 10-membered heteroaryl groups, preferably phenyl, pyrimidinyl and pyridinyl;

[0061] R 5-1 Selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, halogenated C 1-6 Alkyl, cyano, 3- to 8-membered cycloalkyl, -NO2, and amino; preferably C 1-6 Alkoxy, -NR n3 R n4 -CH2-NR n3 R n4 , 3 to 8 membered heterocyclic groups, halogens, cyano groups, 3 to 8 membered cycloalkyl groups and -NO2;

[0062] R n3 and R n4 Each is independently selected from H and C. 1-6 alkyl;

[0063] p is an integer selected from 0, 1, 2, 3.

[0064] On the other hand, this application provides a compound represented by formula (II), a pharmaceutically acceptable salt thereof, and an isomer thereof:

[0065] in,

[0066] R4 is selected from H and C. 1-6 Alkyl groups, preferably CH3;

[0067] Ring A is selected from C6-C 10 Aryl, preferably phenyl;

[0068] R 5-1 Selected from C 1-6 Alkoxy, C 1-6Alkyl group, preferably -OCH3; p is selected from integers of 0, 1, 2, 3.

[0069] In some embodiments of this application, in the compound of formula (II) above, R4 is selected from CH3.

[0070] In some embodiments of this application, the compound of formula (II) above, ring A is selected from phenyl.

[0071] In certain embodiments of this application, the compound of formula (II) above, R 5-1 Selected from -OCH3.

[0072] In some embodiments of this application, the compound of formula (II) above, p is selected from 3.

[0073] On the other hand, this application provides a compound represented by formula (III), a pharmaceutically acceptable salt thereof, and an isomer thereof:

[0074] Where R1 is C 1-6 alkoxy group; R3 is -C(O)C 1-6 Alkyl; the C 1-6 Alkyl groups are replaced by hydroxyl groups;

[0075] Alternatively, R1 can be H and R3 can be -C(O)C. 1-6 alkyl;

[0076] Preferred,

[0077] R1 is C 1-6 alkoxy group; R3 is -C(O)CH2OH;

[0078] Alternatively, R1 can be H and R3 can be -C(O)CH3.

[0079] On the other hand, this application provides a compound of formula (IV-1), its pharmaceutically acceptable salt, and its isomers:

[0080] in,

[0081] R 5-6 Selected from C 1-6 Alkyl groups and -C(=O)-R 5-2 The C 1-6 Alkyl groups are optionally C 1-6 Alkyl substitution;

[0082] R 5-2 Selected from C 1-6 Alkyl, -NH-C 1-6 Alkyl, 3- to 8-membered cycloalkyl, wherein the C 1-6 Alkyl groups may be optionally replaced by hydroxyl or amino groups;

[0083] R 5-3 Selected from C 1-6 Alkyl, the C 1-6 Alkyl groups may be optionally replaced by hydroxyl groups.

[0084] Preferred, R 5-6 for

[0085] R 5-3 Selected from CH3 and CH2OH;

[0086] The optimal choice, R 5-6 for

[0087] On the other hand, this application provides a compound of formula (IV-2), its pharmaceutically acceptable salt, and its isomers:

[0088] in,

[0089] R n1 and R n2 Each is independently selected from H and C. 1-6 Alkyl, the C 1-6 Alkyl groups may be optionally amino- or C-shaped. 1-6 Alkylamino, hydroxyl, 4- to 10-membered bridged cycloalkyl, 3- to 8-membered cycloalkyl, 3- to 8-membered heterocyclic, 5- to 10-membered heteroaryl substitutions, wherein the 4- to 10-membered bridged cycloalkyl, 3- to 8-membered cycloalkyl, 3- to 8-membered heterocyclic, and 5- to 10-membered heteroaryl groups are optionally substituted with amino groups; or,

[0090] R n1 R n2 Together with the nitrogen atoms they are attached to, they form 3- to 8-membered heterocyclic groups, which are optionally C-shaped. 1-6 Alkyl substitution, wherein C 1-6 Alkyl groups may be optionally replaced by amino or hydroxyl groups.

[0091] In some embodiments of this application, the compound of formula (IV-2) above, R n1 Selected from H, R n2 Selected from

[0092] In some embodiments of this application, the compound of formula (IV-2) above, R n1 R n2 Together with the nitrogen atoms they are attached to form

[0093] On the other hand, this application provides a novel anthracycline compound, its pharmaceutically acceptable salt, and its isomers, selected from:

[0094] On the other hand, this application provides an antibody-drug conjugate comprising a small molecule drug, a linker, and an antibody, wherein the small molecule drug is selected from the above-mentioned compounds, their pharmaceutically acceptable salts, and their isomers.

[0095] On the other hand, this application provides the above-mentioned compounds, their pharmaceutically acceptable salts, and their isomers as small molecule drug active ingredients for antibody-drug conjugates.

[0096] On the other hand, this application provides the above-mentioned compounds, their pharmaceutically acceptable salts, their isomers, for the treatment and / or prevention of tumors.

[0097] On the other hand, this application provides a pharmaceutical composition comprising the above-mentioned compound, a pharmaceutically acceptable salt thereof, an isomer thereof, and one or more pharmaceutically acceptable carriers.

[0098] On the other hand, this application provides the use of the above-mentioned compounds, their pharmaceutically acceptable salts, their isomers, or the above-mentioned pharmaceutical compositions in the preparation of medicaments for treating and / or preventing tumors.

[0099] On the other hand, this application provides a method for treating and / or preventing tumors, the method comprising administering an effective amount of the above-mentioned compound, its pharmaceutically acceptable salt, its isomer, or the above-mentioned pharmaceutical composition to an individual in need.

[0100] In some embodiments of this application, the content of the compound, its pharmaceutically acceptable salt, and its isomers in the pharmaceutical composition is 1%-95%.

[0101] In some embodiments of this application, the pharmaceutically acceptable carrier in the pharmaceutical composition includes one or more of fillers, disintegrants, binders, flow aids, and lubricants.

[0102] Terminology Explanation

[0103] Unless otherwise stated, the following terms and phrases used in this application are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense.

[0104] The term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in human and animal tissues to the extent of reasonable medical judgment without excessive toxicity, irritation, allergic reactions, or other problems or complications, and that are commensurate with a reasonable benefit / risk ratio.

[0105] The term "pharmaceutically acceptable salt" refers to a derivative obtained by preparing the compound of this application with a relatively non-toxic acid or base. These salts can be prepared during the synthesis, isolation, and purification of the compound, or by reacting the purified compound in its free form with a suitable acid or base. When the compound contains a relatively acidic functional group (e.g., -COOH, -OH, -SO3H, etc.), it reacts with a suitable inorganic or organic cation (base) to give a base addition salt, including salts formed with alkali metals or alkaline earth metals, ammonium salts formed with amines or their derivatives, and salts formed with amino acids. When the compound contains a relatively basic functional group (e.g., -NH2, etc.), it reacts with a suitable inorganic or organic anion (acid) to give an acid addition salt, including salts formed with inorganic or organic acids (e.g., carboxylic acids, etc.).

[0106] The term "pharmaceutically acceptable carrier" refers to a medium generally acceptable in the art for delivering a bioactive pharmaceutical agent to animals, particularly mammals. Depending on the route of administration and dosage form, this includes, for example, adjuvants, excipients, or excipients such as diluents, preservatives, fillers, flow modifiers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, aromatizers, antibacterial agents, antifungal agents, lubricants, and dispersants. Pharmaceutically acceptable carriers are formulated based on a multitude of factors, within the scope of those skilled in the art. These include, but are not limited to, the type and nature of the formulated active pharmaceutical agent, the recipient to whom the composition containing the pharmaceutical agent is to be administered, the intended route of administration of the composition, and the target therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous media, as well as various solid and semi-solid dosage forms. In addition to the active pharmaceutical agent, such carriers include many different components and additives, and the inclusion of such additional components in the formulation for various reasons (e.g., stabilizing active pharmaceutical agents, binders, etc.) is well known to those skilled in the art.

[0107] The term "effective preventive or therapeutic dose" refers to a sufficient amount of the compound of this application, its pharmaceutically acceptable salts, or isomers thereof, to provide a reasonable benefit / risk ratio for treating any medical condition and / or preventing the disorder. However, it should be understood that the total daily dose of the compound of Formula I of this application, its pharmaceutically acceptable salts, and the composition thereof must be determined by the attending physician within the bounds of reliable medical judgment. For any given patient, the specific effective therapeutic dose level must be determined based on a number of factors, including the disorder being treated and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health condition, sex, and diet; the timing, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors known in the medical field.

[0108] The term "isomer" as used in this application includes geometric isomers and stereoisomers, such as hindered rotation isomers, cis-trans isomers, enantiomers, diastereomers, tautomers, and their racemic mixtures and other mixtures, all of which are within the scope of this application. The term "enantiomer" refers to stereoisomers that are mirror images of each other. The term "tautomer" refers to a functional group isomer that has different hydrogen bonding sites through one or more double bond shifts; for example, a ketone and its enol form are keto-enol tautomers. The term "diastereomer" refers to stereoisomers of molecules having two or more chiral centers and being non-mirror images of each other. The term "cis-trans isomer" refers to different spatial configurations of a molecule where double bonds or single bonds of cyclic carbon atoms cannot rotate freely. The term "hindered rotation isomer" refers to stereoisomers that can be separated due to hindered or very slow single bond rotation. The stereoisomers of the compounds in this application can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. For example, an enantiomer of a compound in this application can be prepared by asymmetric catalysis or chiral derivative derivatization. Alternatively, a single stereoisomer can be obtained from a mixture using chiral resolution techniques. Alternatively, it can be prepared directly from chiral starting materials. The separation of optically pure compounds in this application is typically accomplished using preparative chromatography, employing a chiral column to achieve the separation of chiral compounds.

[0109] The absolute stereoconfiguration of a compound can be confirmed using conventional techniques in the art. For example, single-crystal X-ray diffraction can be used, or the absolute configuration of the compound can be confirmed by examining the chiral structure of the starting materials and the reaction mechanism of asymmetric synthesis. Alternatively, after resolution, the stereoconfiguration can be determined by comparing it with a product whose absolute configuration is known. Compounds marked as "absolute configuration unknown / undetermined" in this application are typically racemic compounds resolved into single isomers via chiral preparative SFC, followed by characterization and testing.

[0110] Those skilled in the art know that when a cyclic compound has a coplanar delocalized system and the number of π electrons is 4n+2, the ring is aromatic. The aromatic structure in a compound can be represented by either dashed lines indicating electron delocalization or by alternating single and double bonds. For example, the structure of a benzene ring can be drawn as follows: It can also be

[0111] The term "optionally substituted" as used in this application refers to two scenarios: one or more hydrogen atoms of the substituted group can be "substituted" or "not substituted" by one or more substituents.

[0112] When the substituent structure contains... A truncated bond indicates that the bond is a linking bond to a substituent, for example... This indicates that the pyrimidine ring is attached to a given group or given structural formula via a carbon atom. A dash "-" in the substituent structure indicates the connection point for the substituent; for example, -SCH3 is attached to a given group or given structural formula via a sulfur atom. The absolute configuration representing the center of a solid, i.e., the R or S configuration. It indicates cis or trans configuration. Double real or double dummy bonds both indicate cis configuration, and one real and one dummy bond indicates trans configuration.

[0113] When a substituent can be cross-bonded to a ring, it means that the substituent can bond with any atom on that ring. For example, structural units. This indicates that the substituent R can be substituted at any position on the benzene ring.

[0114] When a listed substituent does not specify which atom it is attached to a given group or given structural formula, the substituent may be attached by any of its bondable atoms.

[0115] When any variable (e.g., R) d When a compound appears more than once in its composition or structure, its definition is independent in each case. For example, This indicates that the cyclopentyl group is surrounded by 3 R groups. d Replaced, and each R d Each has its own independent options.

[0116] Unless otherwise specified, the term "halogen" refers to a fluorine, chlorine, bromine, or iodine atom.

[0117] Unless otherwise specified, the term "alkyl" refers to a branched or straight-chain saturated aliphatic alkane with a specified number of carbon atoms, minus a hydrogen-derived group. For example, "C 1-10"Alkyl" refers to compounds including C1, C2, C3, C4, C5, C6, C7, C8, C9, C6, C7, C8, C9, C9, C1, C1, C1, C1, C2, C3, C4, C5, C6, C7, C8, C9, C1 ... 10 Alkyl, "C 1-6 Alkyl", C 1- 4-alkyl", C 1-3 Alkyl; specific examples include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, sec-butyl, 2-methylbutyl, 1,1-dimethylbutyl, etc.

[0118] Unless otherwise specified, the term "halogenated alkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by halogen atoms. Preferred halogenated C 1-6 Alkyl, more preferably halogenated C 1-4 Alkyl groups. Examples of haloalkyl groups include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2,2,2-trifluoroethyl, 2,2,2-trichloroethyl, etc. Alkyl groups are as defined above.

[0119] Unless otherwise specified, the term "hydroxyalkyl" refers to a group derived from an alkyl group in which one or more hydrogen atoms are replaced by hydroxyl groups. The term "hydroxyalkyl" as used in this application includes "hydroxyl C..." 1-6 Alkyl group, hydroxyl group 1-4 Alkyl groups; specific examples include, but are not limited to, -CH2OH, -CH2CH2OH, -CH(OH)CH3, and -CH2CH2CH2OH. wait.

[0120] Unless otherwise specified, the term "alkoxy" refers to an alkyl group as defined in this application that is linked to another group by an oxygen atom, i.e., "alkyl-O-". This includes "C". 1-6 Alkoxy (structure is C) 1-6 alkyl-O-), "C 1-4 "Alkoxy" is a suffix, specifically including but not limited to methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, etc.; preferably, the "alkoxy" in this application is preferably C 1-4 Alkoxy, more preferably C 1-3 Alkyl group.

[0121] Unless otherwise specified, the term "haloalkoxy" refers to a group obtained by substituting one or more hydrogen atoms in an alkoxy group with a halogen. Preferably, the "haloalkoxy" described in this application is "haloC". 1-6 Alkoxy, halogenated C 1-4 Alkoxy groups. Specific examples described in this application include: fluoromethoxy groups (including monofluoromethoxy, difluoromethoxy, and trifluoromethoxy), -OCH2CF3, -OCHFCH3, etc. Alkoxy groups are as defined above.

[0122] Unless otherwise specified, the term "cycle" refers to saturated, partially saturated, or unsaturated monocyclic and polycyclic rings, including spirocyclic, fused, or bridged rings. The group derived from a ring by removing a hydrogen atom is called a "cycloyl group," which includes monovalent, divalent (commonly referred to as a subcyclic ring), trivalent, and tetravalent rings, with the specific valence depending on the number of substituents attached to the ring. This application does not specifically distinguish the valence of the ring in its description of "cycloyl groups." Representative "cycloyl groups" include substituted or unsubstituted cycloalkyl, heterocyclic alkyl, cycloalkenyl, heterocyclic alkenyl, cycloynyl, heterocyclic alynyl, aryl, or heteroaryl groups.

[0123] The term "hetero" refers to substituted or unsubstituted heteroatoms and their oxidized forms (also called heteroatomic groups). The heteroatoms are generally selected from N, O, S, and P, and the oxidized forms generally include NO, SO, S(O)2, and P(O). The nitrogen atom may be substituted, i.e., NR (R is H or other substituents defined in the text). The number of atoms on the ring is usually defined as the ring number. For example, "3-6 membered heterocyclic alkyl" refers to a ring consisting of 3-6 atoms arranged in a ring, each ring optionally containing 1 to 3 heteroatoms and / or heteroatomic groups, i.e., N, O, S, NO, SO, S(O)2, P(O), or NR. Each ring is optionally substituted by an R group, R being a group defined in the text.

[0124] Unless otherwise specified, the term "cycloalkyl" refers to a saturated cyclic alkyl group derived from a cycloalkane by removing a hydrogen atom, including monocyclic or polycyclic saturated hydrocarbon groups; the polycyclic saturated hydrocarbon group refers to a polycyclic group formed by two or more cyclic alkyl structures linked by spiro, bridging, fused, or other means. The carbon atom in the cycloalkyl group can be further oxidized, i.e., forming C(O). Unless specifically stated otherwise, "membered cycloalkyl" in this application can be understood as a monocyclic cycloalkyl group, and when it is polycyclic, it will be specifically specified as a spiro, fused, or bridged ring group. The cycloalkyl group includes "3 to 10-membered cycloalkyl", "5 to 10-membered cycloalkyl", "3 to 8-membered cycloalkyl", "4 to 8-membered cycloalkyl", "3 to 6-membered cycloalkyl", and "3 to 5-membered cycloalkyl". Preferably, the cycloalkyl group is a monocyclic, saturated structure, such as C. 3-8 Cycloalkyl; specific examples include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0125] Unless otherwise specified, "cycloalkenyl" means one or more of the cycloalkyl groups in which the cyclic bonds are double bonds and the cycloalkenyl group is not aromatic. The carbon atom in the cycloalkenyl group may be further oxidized, forming C(O). The cycloalkenyl group includes "3- to 10-membered cycloalkenyl", "5- to 10-membered cycloalkenyl", "3- to 8-membered cycloalkenyl", "3- to 6-membered cycloalkenyl", "3- to 5-membered cycloalkenyl", and "5- to 6-membered cycloalkenyl". Specific examples include, but are not limited to, those mentioned above.

[0126] Unless otherwise specified, the term "heterocyclic group" refers to a saturated cyclic group derived by replacing one or more cyclic carbon atoms in a cycloalkyl group with heteroatoms and / or heteroatom groups. The heteroatoms and / or heteroatom groups are generally selected from N, O, S, NO, SO, S(O)₂, P(O), and NR, wherein the carbon atoms in the heterocycle are optionally oxidized to form -C(O); preferably, the heteroatoms are independently selected from 1 to 3 N and / or O atoms. The heterocyclic groups include "3- to 10-membered heterocyclic groups", "5- to 10-membered heterocyclic groups", "3- to 8-membered heterocyclic groups", "3- to 6-membered heterocyclic groups", "3- to 5-membered heterocyclic groups", "4- to 8-membered heterocyclic groups", "4- to 6-membered heterocyclic groups", and "5- to 6-membered heterocyclic groups". Specific examples include, but are not limited to, nitrogen-containing heterocyclic butyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, etc.

[0127] Unless otherwise specified, the term "heterocyclic alkenyl" refers to a "heterocyclic group" in which one or more cyclic bonds are double bonds and the heterocycle is not aromatic. Preferably, the heteroatoms are independently selected from 1-3 N and / or O atoms. The heterocyclic group includes "3-8-membered heterocyclic alkenyl", "3-6-membered heterocyclic alkenyl", "3-5-membered heterocyclic alkenyl", and "5-6-membered heterocyclic alkenyl". Specific examples include, but are not limited to: wait.

[0128] Unless otherwise specified, the term "aryl" refers to an unsaturated, usually aromatic, hydrocarbon group, which may be a monocyclic or a plurality of rings fused together. Preferably, it is a 5- to 10-membered aryl group, more preferably a 5- to 8-membered aryl group, and most preferably a monocyclic 5- to 6-membered aryl group; examples of aryl groups include, but are not limited to, phenyl and naphthyl groups.

[0129] The term "heteroaryl" as used in this application refers to an aromatic monocyclic or polycyclic group containing one or more heteroatoms, wherein the heteroatoms are generally selected from N, O, and S; preferably, the heteroatoms are independently selected from 1-3 N and / or O atoms, and the N and S atoms may optionally be oxidized and the N atom may optionally be quaternized. The term "heteroaryl" includes "monocyclic heteroaryl" and "fused-ring heteroaryl," wherein a fused-ring heteroaryl refers to an aromatic group consisting of one or more heteroatoms formed by two or more cyclic structures sharing two adjacent atoms. Unless otherwise specified, the term "heteroaryl" as used in this application can generally be understood as "monocyclic heteroaryl," such as the "5-6 membered heteroaryl" mentioned in this application, which does not have the potential to form a fused-ring heteroaryl; when it is a "fused-ring heteroaryl," it will be specifically indicated as a "fused" heteroaryl structure, such as "8-10 membered fused-ring heteroaryl." The heteroaryl group described in this application is preferably a "nitrogen-containing heteroaryl group," more preferably a "5-6 member nitrogen-containing aryl group." The heteroatom in the "nitrogen-containing heteroaryl group" contains at least one nitrogen atom, for example, only one, two, or three nitrogen atoms; or, it contains one nitrogen atom and one or two other heteroatoms (e.g., S and / or O atoms); or, it contains two nitrogen atoms and one or two other heteroatoms. Specific examples of the heteroaryl group include, but are not limited to: furanyl, thiopheneyl, pyrroleyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazoleyl, pyrazolyl, etc.

[0130] Unless otherwise specified, the term "bridged ring group" refers to a saturated or partially saturated ring structure formed by two or more carbon rings sharing two non-adjacent carbon atoms. Optionally, the carbon atoms in the ring structure may be substituted with oxygen. "Bridged ring group" includes, for example, "4-10-membered bridged ring group," "5-11-membered bridged ring group," "6-11-membered bridged ring group," "5-10-membered bridged ring group," "5-7-membered bridged ring group," "7-10-membered bridged ring group," "6-9-membered bridged ring group," "7-8-membered bridged ring group," "9-10-membered bridged ring group," etc. Specific examples include, but are not limited to:

[0131] Unless otherwise specified, the term "bridged heterocyclic group" refers to a saturated or partially saturated cyclic structure derived from the substitution of at least one carbon atom in the "bridged cyclic group" by a heteroatom / heteroatomic group selected from N, O, S, P, NO, SO, S(O)2, P(O), and NR, where R is H or any possible substituent group, and the "bridged cycloalkyl group" is as defined above. "Bridged heterocyclic groups" include, for example, "4-10-membered bridged heterocyclic groups," "5-11-membered bridged heterocyclic groups," "6-11-membered bridged heterocyclic groups," "5-10-membered bridged heterocyclic groups," "7-10-membered bridged heterocyclic groups," "6-9-membered bridged heterocyclic groups," "7-8-membered bridged heterocyclic groups," "9-10-membered heterobridged cyclic groups," "6-10-membered bridged heterocyclic alkenyl groups," "6-8-membered bridged heterocyclic alkenyl groups," and "7-8-membered nitrogen-containing bridged heterocyclic alkenyl groups," etc. Preferably, the heteroatoms are independently selected from 1-3 N and / or O atoms. Preferably, the bridging heterocyclic group is a "nitrogen-containing bridging heterocyclic group," meaning that at least one ring atom is N, and optionally contains one or more other heteroatoms; preferably, the "nitrogen-containing bridging heterocyclic group" contains one N atom and 0-2 atoms selected from N and / or O and / or S. Preferably, the "nitrogen-containing bridging heterocyclic group" contains one N atom and 0-1 atoms selected from O and / or S. Preferably, the bridging heterocyclic group is an "oxygen-containing bridging heterocyclic group," meaning that at least one ring atom is O, and optionally contains one or more other heteroatoms; preferably, the "oxygen-containing heterocyclic group" contains one O atom and 0-2 atoms selected from N and / or O. Specific examples include, but are not limited to: wait.

[0132] When a substituent can be cross-bonded to a ring, it means that the substituent can bond with any atom on that ring. For example, structural units. Indicates substituent R b Substitution can occur at any position on ring B (including NH), and R b The number of atoms is n; when the ring contains NH, it means that NH, like other ring atoms, can be converted by R. b What it replaces, namely R b It can replace H, or it can choose not to replace H.

[0133] When any variable (e.g., R) b When a structural unit appears more than once in the composition or structure of a compound, its definition is independent in each case. For example, a structural unit. This indicates that ring B is divided by n R's. b Replaced, and each R b Each has its own independent options.

[0134] When a listed substituent does not specify which atom it is attached to a given group or given structural formula, the substituent may be attached by any of its bondable atoms. For example, pyrimidine as a substituent means that any carbon or nitrogen atom on the pyrimidine ring is attached to the substituted group. Another example is ring B in this application, selected from... The absence of a defined connection site in the structural unit means that any carbon atom on the tetrahydropyran ring can be attached to a given group or a given structural formula.

[0135] In particular, any combination of substituents and / or their variants in this application is permitted only if such a combination produces a stable compound.

[0136] "Substitution" or "substituted" refers to one or more hydrogen atoms in a group, preferably 1 to 6, more preferably 1 to 3 hydrogen atoms, which are independently substituted by the corresponding number of substituents. Those skilled in the art can determine possible or impossible substitutions without much effort (through experimentation or theory). For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond (such as an alkene).

[0137] "Pharmaceutical composition" refers to a mixture containing one or more of the compounds described in this application or their physiologically / pharmacologically acceptable salts or prodrugs, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and enabling it to exert its biological activity.

[0138] The term "medicinal salt" refers to derivatives obtained from the compounds of this application prepared with relatively non-toxic acids or bases. These salts can be prepared during the synthesis, isolation, and purification of the compounds, or by reacting the purified free form of the compounds with suitable acids or bases. When the compounds contain relatively acidic functional groups, they react with alkali metal, alkaline earth metal hydroxides, or organic amines to yield base addition salts, including cations based on alkali metals and alkaline earth metals, as well as non-toxic ammonium, quaternary ammonium, and amine cations, and also encompassing amino acid salts. When the compounds contain relatively basic functional groups, they react with organic or inorganic acids to yield acid addition salts.

[0139] As used in this application, the singular forms of “a,” “an,” and “the” include plural references, and vice versa, unless the context clearly indicates otherwise.

[0140] When the term "about" is applied to parameters such as pH, concentration, temperature, etc., it indicates that the parameter can vary by ±10%, and sometimes more preferably within ±5%. As those skilled in the art will understand, when a parameter is not critical, figures are usually given for illustrative purposes only, not as limitations.

[0141] The control loading molecule Dxd in this application is a topoisomerase I inhibitor and has the following structural formula.

[0142] The preparation methods for some compounds in this application reference the preparation methods for the aforementioned similar compounds. Those skilled in the art should understand that when using or referring to the referenced preparation methods, the reactant ratios, reaction solvents, reaction temperatures, etc., can be appropriately adjusted according to the different reactants.

[0143] The compounds of this application can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this application.

[0144] Summary of experimental instruments:

[0145] The structures of the compounds in this application were determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS), or ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR determinations were performed using a Bruker Neo 400M or Bruker Ascend 400 NMR spectrometer, with solvents including deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3), heavy water (D2O), and tetramethylsilane (TMS) as the internal standard.

[0146] The determination was performed using an Agilent 1260-6125B single quadrupole mass spectrometer with a Welch Biomate column (C18, 2.7 μm, 4.6 × 50 mm) or a Waters H-Class SQD2 column with a Welch Ultimate column (XB-C18, 1.8 μm, 2.1 × 50 mm) (ion source: electrospray ionization).

[0147] The determination was performed using ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) with a Waters UPLC H-class SQD mass spectrometer (ion source: electrospray ionization).

[0148] HPLC determinations were performed using a Waters e2695-2998 or Waters ARC and an Agilent 1260 or Agilent Poroshell HPH high-performance liquid chromatograph.

[0149] Preparative HPLC was performed using a Waters 2555-2489 (10 μm, ODS 250 cm × 5 cm) or a GILSON Trilution LC column, with a Welch XB-C18 column (5 μm, 21.2 × 150 mm).

[0150] Thin-layer chromatography (TLC) uses GF254 silica gel plates from Yantai Jiangyou Silica Gel Development Co., Ltd. or GF254 silica gel plates from Rushan Shangbang New Materials Co., Ltd. The TLC standard is 0.15mm to 0.20mm, with a preparative type of 20×20cm. Column chromatography generally uses 200-300 mesh silica gel from chemical industry as the carrier. Detailed Implementation

[0151] The starting materials used in the embodiments of this application are known and commercially available, or can be synthesized using methods known in the art. Unless otherwise specified, all reactions in this application are carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere, using a dry solvent, and the reaction temperature is expressed in degrees Celsius or °C. The room temperature generally refers to 15-35 °C, preferably 20-30 °C, and more preferably 20-25 °C. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of this application without departing from the spirit and scope thereof.

[0152] Example 1

[0153] Synthesis of (8S,10S)-6,8,11-trihydroxy-10-(((2R,4S,5S,6S)-5-hydroxy-6-methyl-4-(methyl(3,4,5-trimethoxybenzyl)amino))tetrahydro-2H-pyran-2-yl)oxy)-8-(2-hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione (compound A-3)

[0154] Step 1:

[0155] Synthesis of (8S,10S)-6,8,11-trihydroxy-10-(((2R,4S,5S,6S)-5-hydroxy-6-methyl-4-((3,4,5-trimethoxybenzyl)amino)tetrahydro-2H-pyran-2-yl)oxy)-8-(2-hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione (compounds 1-2)

[0156] At room temperature, doxorubicin hydrochloride (compound 1-1, 290.0 mg, 0.5 mmol) was dissolved in a mixed solvent of acetonitrile and water (3:1, 16 mL). 3,4,5-trimethoxybenzaldehyde (981.0 mg, 5.0 mmol) was then added to the solution, and the mixture was stirred at room temperature for 30 minutes. Sodium cyanoborohydride (62.8 mg, 1.0 mmol) was then added to the reaction system, and the mixture was stirred at room temperature for another 30 minutes. The reaction was monitored by LC-MS. 8 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The remaining red solid was used directly in the next reaction step.

[0157] Step Two:

[0158] Synthesis of compound A-3

[0159] At room temperature, the residual solid obtained in step one was dissolved in ethanol (18.0 mL), followed by 37 wt% formaldehyde aqueous solution (1.0 mL, 13.4 mmol) and sodium cyanoborohydride (50 mg, 0.8 mmol). The mixture was stirred at room temperature for 45 minutes, and the reaction was monitored by LC-MS. 8 mL of water was added to the reaction mixture, and the mixture was extracted with chloroform (15 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 96:4) to give the brownish-red title compound (82 mg).

[0160] LC-MS: 738.2 [M+H] + ;

[0161] 1 H NMR (400MHz, Chloroform-d): δ14.01(s,1H),13.28(s,1H),8.05(dd,J=7.7,1.0Hz,1H),7.80(dd,J=8.5,7.7Hz,1H ),7.41(dd,J=8.6,1.1Hz,1H),6.46(s,2H),5.61(s,1H),5.34(s,1H),4.76(s,3H),4.09(s,3H),3.99(q,J=6.7Hz, 1H),3.80(d,J=8.8Hz,9H),3.69–3.55(m,1H),3.43(s,0H),3.29(dd,J=18.9,2.0Hz,1H),3.06(d,J=18.9Hz,1H),3 .01(s,1H),2.61(s,1H),2.39(d,J=14.8Hz,1H),2.18(dd,J=14.7,4.0Hz,1H),1.92(s,1H),1.40(d,J=6.5Hz,3H).

[0162] Example 2

[0163] Synthesis of (8S,10S)-10-(((2R,4S,5S,6S)-4-((4-aminobenzyl)(methyl)amino)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,8,11-trihydroxy-8-(2-hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione (compound A-6)

[0164] Step 1:

[0165] Synthesis of tert-butyl(4-((((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)tetrahydro-2H-pyran-4-yl)amino)methyl)phenyl)carbamate (compound 2-1))

[0166] At room temperature, doxorubicin hydrochloride (compound 1-1, 290.0 mg, 0.5 mmol) was dissolved in a mixture of acetonitrile and water (3:1, 16 mL). 4-(Boc-amino)benzaldehyde (1.1 g, 5.0 mmol) was then added to the solution, and the mixture was stirred at room temperature for 30 minutes. Sodium cyanoborohydride (62.8 mg, 1.0 mmol) was then added to the reaction mixture, and the mixture was stirred at room temperature for another 30 minutes. The reaction was monitored by LC-MS. 8 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The remaining red solid was used directly in the next reaction step.

[0167] Step Two:

[0168] Synthesis of tert-butyl (4-((((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)tetrahydro-2H-pyran-4-yl)(methyl)amino)methyl)benzene)carbamate (compound 2-2)

[0169] At room temperature, the residual solid obtained in step one was dissolved in ethanol (18 mL), followed by 37 wt% formaldehyde aqueous solution (1.0 mL, 13.4 mmol) and sodium cyanoborohydride (50 mg, 0.8 mmol). The mixture was stirred at room temperature for 45 minutes, and the reaction was monitored by LC-MS. 8 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give the brownish-red title compound (98 mg).

[0170] Step 3:

[0171] Synthesis of compound A-6

[0172] At room temperature, zinc bromide (337.8 mg, 1.5 mmol) was added to a solution of compound 2-2 (76.3 mg, 0.1 mmol) in dichloromethane (3 mL), and the reaction was carried out at room temperature for 5 hours, monitored by LC-MS. 5 mL of water was added, and the mixture was stirred for 5 minutes. Then, 5 mL of saturated sodium bicarbonate solution was added, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 93:7) to give the brownish-red title compound (41 mg).

[0173] LC-MS: 663.2 [M+H] + ;

[0174] 1 H NMR (400MHz, Chloroform-d): δ13.99(s,1H),13.29(s,1H),8.05(d,J=7.6Hz,1H),7.79(t,J=8.1Hz,1H),7.40(d, J=8.5Hz,1H),6.99(d,J=8.0Hz,2H),6.60(d,J=8.1Hz,2H),5.59(s,1H),5.34(s,1H),4.78(d,J=7.1Hz,3H),4.09 (s,3H),3.98(q,J=6.6Hz,1H),3.83(s,1H),3.75–3.48(m,1H),3.38–3.20(m,2H),3.06(d,J=18.8Hz,1H),2.63–2 .50(m,3H),2.39(d,J=14.6Hz,1H),2.17(dd,J=14.7,3.9Hz,1H),2.10(s,3H),1.87(s,2H),1.41(d,J=6.5Hz,3H).

[0175] Example 3

[0176] Synthesis of (8S,10S)-6,8,11-trihydroxy-10-(((2R,4S,5S,6S)-5-hydroxy-6-methyl-4-(((2-morpholinopyrimidin-5-yl)methyl)amino)tetrahydro-2H-pyran-2-yl)oxy)-8-(2-hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione (compound A-15)

[0177] Step 1:

[0178] Synthesis of 2-chloro-5-(dimethoxymethyl)pyrimidine (compound 3-2)

[0179] 2-Chloroprene-5-carboxaldehyde (compound 3-1, 306 mg, 2.15 mmol) was dissolved in 6 mL of dry 1,4-dioxane at room temperature. Trimethyl orthoformate (1 mL, 9.7 mmol) and p-toluenesulfonic acid (38 mg, 0.22 mmol) were then added to the solution. The reaction mixture was refluxed at 85 °C for 1 hour. After cooling to room temperature, the reaction flask was placed in an ice bath, and the reaction was quenched with 1 mL of saturated sodium bicarbonate solution. The mixture was then diluted with 2 mL of water, extracted with dichloromethane (10 mL × 3), and the combined organic phases were dried over anhydrous sodium sulfate. The mixture was then filtered, concentrated under reduced pressure, and the resulting oily product was used directly in the next reaction step.

[0180] Step Two:

[0181] Synthesis of 4-(5-(dimethoxymethyl)pyrimidin-2-yl)morpholine (compound 3-3)

[0182] Compound 3-2 (394 mg, 2.1 mmol) was dissolved in 12 mL of DMF at room temperature, followed by the sequential addition of morpholine hydrochloride (319 mg, 2.58 mmol) and triethylamine (600 μl, 4.3 mmol). The reaction mixture was incubated at 90 °C for 1.5 h, and the reaction was monitored by TLC. The mixture was concentrated under reduced pressure, and the residual solid was purified by rapid silica gel column chromatography (ethyl acetate / petroleum ether, 1:20) to give 430 mg of a white solid, which was the title compound.

[0183] Step 3:

[0184] Synthesis of 2-morpholinopyrimidine-5-carboxaldehyde (compounds 3-4)

[0185] Compound 3-3 (430 mg, 1.8 mmol) was dissolved in 20 mL of tetrahydrofuran, followed by the addition of water (1 mL) and p-toluenesulfonic acid (31 mg, 0.18 mmol). The mixture was heated under reflux for 3 hours, and the reaction was monitored by TLC. The reaction mixture was cooled to room temperature, diluted with 5 mL of water, and extracted with dichloromethane (15 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate. The mixture was then filtered, concentrated under reduced pressure, and the residual solid was purified by rapid silica gel column chromatography (petroleum ether / ethyl acetate, 20:1) to give 360 ​​mg of a pale yellow solid, which was the title compound.

[0186] Step Four:

[0187] Synthesis of (8S,10S)-6,8,11-trihydroxy-10-(((2R,4S,5S,6S)-5-hydroxy-6-methyl-4-(((2-morpholinopyrimidin-5-yl)methyl)amino)tetrahydro-2H-pyran-2-yl)oxy)-8-(2-hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione (compounds 3-5)

[0188] At room temperature, doxorubicin hydrochloride (compound 1-1, 200.0 mg, 0.373 mmol) was dissolved in a mixture of methanol and water (3:1, 16 mL). Compound 3-4 (360 mg, 5.0 mmol) was then added to the above solution. The mixture was stirred at room temperature for 30 minutes, and then sodium cyanoborohydride (46 mg, 0.75 mmol) was added to the reaction system. The mixture was stirred at room temperature for another hour, and the reaction was monitored by LC-MS. 8 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residual red solid was used directly in the next step of the reaction.

[0189] Step 5:

[0190] Synthesis of compound A-15

[0191] At room temperature, the crude product of compounds 3-5 from the previous step was dissolved in methanol (15 mL), and 37 wt% formaldehyde aqueous solution (1.0 mL, 13.4 mmol) was added. The mixture was stirred for 30 minutes, and then sodium cyanoborohydride (26 mg, 0.41 mmol) was added. The mixture was stirred at room temperature for 25 minutes, and the reaction was monitored by LC-MS. 8 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give 28 mg of the brownish-red title compound.

[0192] LC-MS: 735.3 [M+H] + ;

[0193] 1 H NMR (400MHz, Chloroform-d): δ14.02(d,J=1.9Hz,1H),13.27(d,J=2.3Hz,1H),8.18(s,2H),8.04(dd,J=7.7,1.0Hz,1H),7. 80(t,J=8.1Hz,1H),7.41(dd,J=8.6,1.1Hz,1H),5.59(d,J=3.4Hz,1H),5.33(t,J=2.9Hz,1H),4.76(s,2H),4.09(s,3H),3. 99(dd,J=6.6,1.3Hz,1H),3.89–3.79(m,1H),3.74(s,9H),3.50(d,J=13.4Hz,1H),3.39(s,1H),3.33–3.23(m,1H),3.05(d, J=18.9Hz,1H),2.59(s,1H),2.40(dt,J=14.7,2.2Hz,1H),2.26–2.05(m,4H),1.92(d,J=7.2Hz,2H),1.39(d,J=6.6Hz,3H).

[0194] Example 4

[0195] Synthesis of (8S,10S)-10-(((2R,4S,5S,6S)-4-(((2-(dimethylamino)-4-methoxypyrimidin-5-yl)methyl)(methyl)amino)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,8,11-trihydroxy-8-(2-hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione (compound A-18)

[0196] Step 1:

[0197] Synthesis of (2-(dimethylamino)-4-methoxypyrimidin-5-yl)methanol (compound 4-2)

[0198] At room temperature, diethylamine (1.2 mL, 11.50 mmol) and DIPEA (3.0 mL, 17.25 mmol) were added sequentially to a 30 mL solution of dioxane (1.0 g, 5.75 mmol) of compound 4-1. The mixture was heated to 65 °C and refluxed with stirring for 1.5 h. The reaction was monitored by TLC. The solution was concentrated under reduced pressure, and the residual solid was purified by rapid silica gel column chromatography (petroleum ether / ethyl acetate, 10:1) to give 978 mg of white solid, which is the title compound.

[0199] Step Two:

[0200] Synthesis of 2-(dimethylamino)-4-methoxypyrimidine-5-carboxaldehyde (compound 4-3)

[0201] Under ice bath and nitrogen protection, Dess-Martin periodinane (6.86 g, 16.2 mmol) was added in portions to a solution of compound 4-2 (978 mg, 5.34 mmol) in dichloromethane (80 mL). The mixture was stirred for 5 min, and then allowed to react at room temperature for 2 h. The reaction mixture was filtered, the filter cake was washed with methyl tert-butyl ether, the filtrates were combined, concentrated, and the residue was purified by rapid silica gel column chromatography (petroleum ether / ethyl acetate, 15:1) to give 861 mg of the white solid, which is the title compound.

[0202] Step 3:

[0203] Synthesis of (8S,10S)-10-(((2R,4S,5S,6S)-4-(((2-(dimethylamino)-4-methoxypyrimidin-5-yl)methyl)amino)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,8,11-trihydroxy-8-(2-hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione (compound 4-4)

[0204] At room temperature, doxorubicin hydrochloride (compound 1-1, 200.0 mg, 0.373 mmol) was dissolved in a mixed solvent of methanol and water (3:1, 16 mL). Compound 4-3 (675 mg, 3.73 mmol) was then added to the above solution. The mixture was stirred at room temperature for 30 minutes, and then sodium cyanoborohydride (34.5 mg, 0.56 mmol) was added to the reaction system. The mixture was stirred at room temperature for another 3 hours, and the reaction was monitored by LC-MS. 8 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (30 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by rapid silica gel column chromatography (dichloromethane / methanol, 94:6) to give 127 mg of the red solid, which is the title compound.

[0205] Step Four:

[0206] Synthesis of compound A-18

[0207] At room temperature, compound 4-4 (127 mg, 0.18 mmol) from the previous step was dissolved in methanol (15 mL), and 37 wt% formaldehyde aqueous solution (0.5 mL, 6.7 mmol) was added. The mixture was stirred for 30 minutes, followed by the addition of sodium cyanoborohydride (13.6 mg, 0.22 mmol). The mixture was stirred at room temperature for 45 minutes, and the reaction was monitored by LC-MS. 8 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give 43 mg of the brownish-red title compound.

[0208] LC-MS: 723.3 [M+H] + ;

[0209] 1 H NMR(400MHz,Chloroform-d)δ14.00(s,1H),13.28(s,1H),8.05(dd,J=7.7,1.1Hz,1H),7.91(s,1H),7. 80(t,J=8.1Hz,1H),7.40(dd,J=8.6,1.1Hz,1H),5.60(d,J=2.9Hz,1H),5.33(s,1H),4.76(s,2H),4.09 (s,3H),4.00(q,J=6.6Hz,1H),3.86(s,3H),3.67(s,1H),3.29(dd,J=18.9,2.0Hz,1H),3.13(s,6H),3. 09(m,2H),3.02(m,1H),2.39(m,1H),2.17(dd,J=14.8,4.1Hz,1H),1.87(s,1H),1.39(d,J=6.5Hz,3H).

[0210] Example 5

[0211] Synthesis of 5-((((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)tetrahydro-2H-pyran-4-yl)(methyl)amino)methyl)-4-methoxypyrimidine-2-nitrile (compound A-19)

[0212] Step 1:

[0213] Synthesis of 5-(((tert-butyldimethylsilyl)oxy)methyl)-2-chloro-4-methoxypyrimidine (compound 5-1)

[0214] At room temperature, tert-butyldimethylchlorosilane (1.36 g, 9.0 mmol) and imidazole (1.28 g, 18.75 mmol) were added sequentially to a solution of compound 4-1 (1.3 g, 7.5 mmol) in N,N-dimethylformamide (30 mL), and the mixture was stirred for 4 hours. The reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure, diluted with water (30 mL), and extracted with ethyl acetate (30 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure, and the remaining pale yellow oily residue was the crude product of compound 5-1, which could be used directly in the next reaction without purification.

[0215] Step Two:

[0216] The synthesis of 5-(((tert-butyldimethylsilyl)oxy)methyl)-4-methoxypyrimidin-2-onitrile (compound 5-2) was carried out at room temperature. Compound 5-1 (2.02 g, 7.0 mmol) was dissolved in N,N-dimethylformamide (30 mL), followed by the sequential addition of NaCN (1.03 g, 21.0 mmol) and DABCO (3.14 g, 28.0 mmol). The mixture was stirred at this temperature for 48 hours, and the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure, diluted with water (30 mL), and extracted with ethyl acetate (30 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure and purified by rapid silica gel column chromatography (petroleum ether / ethyl acetate, 15:1) to give 1.43 g of a yellow solid, which is the title compound.

[0217] Step 3:

[0218] Synthesis of 5-(hydroxymethyl)-4-methoxypyrimidine-2-nitrile (compound 5-3)

[0219] At room temperature, TBAF (1 M in THF, 6.0 mL, 6.0 mmol) was added dropwise to a tetrahydrofuran (30 mL) solution of compound 5-2 (1.43 g, 5.02 mmol). The reaction mixture was stirred at room temperature for 24 hours, and the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure, diluted with water (30 mL), and extracted with ethyl acetate (30 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure and purified by rapid silica gel column chromatography (petroleum ether / ethyl acetate, 8:1) to give 792 mg of a yellow solid, which is the title compound.

[0220] Step Four:

[0221] Synthesis of 5-formyl-4-methoxypyrimidine-2-nitrile (compound 5-4)

[0222] At room temperature, manganese dioxide (3.34 g, 38.4 mmol) was added in portions to a 20 mL solution of compound 5-3 (792 mg, 4.8 mmol) in tetrahydrofuran. The mixture was heated to 60 °C and stirred for 18 hours, with the reaction monitored by TLC. The reaction mixture was filtered, the filter cake was washed with ethyl acetate, the filtrates were combined, concentrated, and the residue was purified by rapid silica gel column chromatography (petroleum ether / ethyl acetate, 10:1) to give 703 mg of the pale yellow solid, which is the title compound.

[0223] Step 5:

[0224] Synthesis of 5-((((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)tetrahydro-2H-pyran-4-yl)amino)methyl)-4-methoxypyrimidine-2-nitrile (compound 5-5)

[0225] At room temperature, doxorubicin hydrochloride (compound 1-1, 249.4 mg, 0.43 mmol) was dissolved in a mixed solvent of acetonitrile and water (3:1, 18 mL). Compound 5-4 (703 mg, 4.31 mmol) was then added to the above solution. The mixture was stirred at room temperature for 30 minutes, followed by the addition of sodium cyanoborohydride (40.5 mg, 0.65 mmol). The mixture was stirred at room temperature for another 1.5 hours, and the reaction was monitored by LC-MS. 10 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (30 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by rapid silica gel column chromatography (dichloromethane / methanol, 94:6) to give 262 mg of the red solid, which is the title compound.

[0226] Step Six:

[0227] Synthesis of compound A-19

[0228] Compound 5-5 (262 mg, 0.38 mmol) was dissolved in methanol (18 mL) at room temperature. 37 wt% formaldehyde aqueous solution (1.0 mL, 13.4 mmol) was added, and the mixture was stirred for 30 minutes. Sodium cyanoborohydride (28.6 mg, 0.46 mmol) was then added, and the mixture was stirred at room temperature for 45 minutes. The reaction was monitored by LC-MS. 10 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give 138 mg of the brownish-red title compound.

[0229] LC-MS: 705.3 [M+H]+ ;

[0230] 1 H NMR(400MHz,Chloroform-d)δ14.02(s,1H),13.28(s,1H),8.42(s,1H),8.05(d,J=7.7Hz,1H),7.81(t ,J=8.1Hz,1H),7.41(d,J=8.6Hz,1H),5.61(s,1H),5.33(s,1H),4.76(s,2H),4.73(s,1H),4.09(s,3H) ,4.04(s,3H),3.80(d,J=51.3Hz,2H),3.47(s,1H),3.30(d,J=18.8Hz,1H),3.15–2.96(m,2H),2.62(s ,1H),2.39(d,J=14.7Hz,1H),2.19(d,J=14.2Hz,3H),1.95(m,1H),1.88(m,1H),1.39(d,J=6.4Hz,3H).

[0231] Example 6

[0232] Synthesis of (8S,10S)-8-acetyl-10-(((2R,4S,5S,6S)-4-((5-amino-2-methoxybenzyl)(methyl)amino)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,8,11-trihydroxy-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione (compound A-31)

[0233] Step 1:

[0234] Synthesis of tert-butyl (3-(hydroxymethyl)-4-methoxyphenyl)carbamate (compound 6-2)

[0235] At room temperature, compound 6-1 (1.0 g, 4.63 mmol), Boc-NH2 (969 mg, 8.3 mmol), anhydrous Cs2CO3 (3.02 g, 9.26 mmol), Xantphos (805.2 mg, 1.39 mmol), and Pd2(dba)3 (720.7 mg, 0.79 mmol) were added sequentially to a round-bottom flask, followed by anhydrous toluene (20 mL). The mixture was purged with nitrogen three times, and the reaction mixture was heated to 110 °C and stirred for 18 hours. The reaction was monitored by TLC. After cooling to room temperature, the mixture was diluted with ethyl acetate (40 mL), and the particulate matter was removed by diatomaceous earth filtration. The filtrate was concentrated under reduced pressure, and the residue was purified by rapid silica gel column chromatography (petroleum ether / ethyl acetate, 8:1) to give 766 mg of a pale yellow solid, which was the title compound.

[0236] Step Two:

[0237] Synthesis of tert-butyl 3-formyl-4-methoxyphenyl)carbamate (compound 6-3)

[0238] At room temperature, manganese dioxide (2.12 g, 24.4 mmol) was added in portions to a tetrahydrofuran (16 mL) solution of compound 6-2 (766 mg, 3.05 mmol). The mixture was heated to 60 °C and stirred for 18 hours, with the reaction monitored by TLC. The reaction mixture was filtered, the filter cake was washed with ethyl acetate, the filtrates were combined, concentrated, and the residue was purified by rapid silica gel column chromatography (petroleum ether / ethyl acetate, 10:1) to give 612 mg of the pale yellow solid, which is the title compound.

[0239] Step 3:

[0240] Synthesis of tert-butyl(3-((((2S,3S,4S,6R)-6-(((1S,3S)-3-acetyl-3,5,12-trihydroxy-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)-3-hydroxy-2-methyltetrahydro-2H-pyran-4-yl)amino)methyl)-4-methoxyphenyl)carbamate (compound 6-4)

[0241] At room temperature, daunorubicin hydrochloride (174.8 mg, 0.31 mmol) was dissolved in a mixture of acetonitrile and water (3:1, 16 mL). Compound 6-3 (612 mg, 2.44 mmol) was then added to the above solution, and the mixture was stirred at room temperature for 30 minutes. Sodium cyanoborohydride (29.2 mg, 0.47 mmol) was then added to the reaction system, and the mixture was stirred at room temperature for another 3 hours. The reaction was monitored by LC-MS. 10 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by rapid silica gel column chromatography (dichloromethane / methanol, 94:6) to give 153 mg of the red solid, which is the title compound.

[0242] Step Four:

[0243] Synthesis of tert-butyl(3-((((2S,3S,4S,6R)-6-(((1S,3S)-3-acetyl-3,5,12-trihydroxy-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)-3-hydroxy-2-methyltetrahydro-2H-pyran-4-yl)(methyl)amino)methyl)-4-methoxyphenyl)carbamate (compounds 6-5)

[0244] Compound 6-4 (153 mg, 0.2 mmol) was dissolved in methanol (15 mL) at room temperature. 37 wt% formaldehyde aqueous solution (0.5 mL, 6.7 mmol) was added, and the mixture was stirred for 30 minutes. Sodium cyanoborohydride (15.1 mg, 0.24 mmol) was then added, and the mixture was stirred at room temperature for 45 minutes. The reaction was monitored by LC-MS. 8 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give 101 mg of the brownish-red title compound.

[0245] Step 5:

[0246] Synthesis of compound A-31

[0247] Zinc bromide (438.8 mg, 1.95 mmol) was added in portions to a solution of compound 6-5 (101 mg, 0.13 mmol) in dichloromethane (8 mL) at room temperature. The reaction was allowed to proceed for 18 hours at room temperature, and the reaction was monitored by LC-MS. 4 mL of water was added, and the mixture was stirred for 5 minutes. Then, 12 mL of saturated sodium bicarbonate solution was added, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 93:7) to give 69 mg of the brownish-red title compound.

[0248] LC-MS: 693.3 [M+H] + ;

[0249] 1 H NMR(400MHz,Chloroform-d)δ14.02(s,1H),13.28(s,1H),8.43(s,1H),8.05(d,J=7.7Hz,1H),7.81(t, J=8.1Hz,1H),7.41(d,J=8.6Hz,1H),5.61(s,1H),5.33(s,1H),4.77(s,2H),4.73(s,1H),4.09(s,3H),4 .04(m,4H),3.80(d,J=52.6Hz,2H),3.47(s,0H),3.30(d,J=18.8Hz,1H),3.06(d,J=19.8Hz,1H),3.02(s ,1H),2.62(s,1H),2.39(d,J=14.7Hz,1H),2.28-2.10(m,3H),2.04-1.82(m,2H),1.39(d,J=6.4Hz,3H).

[0250] Example 7

[0251] (2S,4S)-2,5,12-trihydroxy-4-(((2R,4S,5S,6S)-5-hydroxy-6-methyl-4-(methyl(3,4,5-trimethoxybenzyl)amino)tetrahydro-2H-pyran-2-yl)oxy)-N-(2-hydroxyethyl)-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-2-carboxamide (Compound A-49)

[0252] Synthesis of (2S,4S)-2,5,12-trihydroxy-4-(((2R,4S,5S,6S)-5-hydroxy-6-methyl-4-(methyl(3,4,5-trimethoxybenzyl)amino)tetrahydro-2H-pyran-2-yl)oxy)-7-methoxy-N-(2-(methylamino)ethyl)-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-2-carboxamide (compound A-51)

[0253] Step 1:

[0254] Synthesis of (2S,4S)-2,5,12-trihydroxy-4-(((2R,4S,5S,6S)-5-hydroxy-6-methyl-4-(methyl(3,4,5-trimethoxybenzyl)amino)tetrahydro-2H-pyran-2-yl)oxy)-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetrabenzo-2-carboxylic acid (compound 10-1)

[0255] At room temperature, compound A-3 (150 mg, 0.203 mmol) was dissolved in a methanol / water mixture (3:2, 30 mL). Then, while stirring, an aqueous solution of sodium periodate (65.1 mg, 0.305 mmol) (6 mL) was added dropwise to the solution. The reaction was continued for 30 minutes at room temperature until complete. The reaction solution was extracted with dichloromethane (30 mL × 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was used directly as the crude product in the next reaction step.

[0256] Step Two:

[0257] Synthesis of compound A-49

[0258] At room temperature, compound 10-1 (144.6 mg, 0.2 mmol) and HATU (155.9 mg, 0.41 mmol) were dissolved in a mixed solvent of DMF and dichloromethane (1:4, 5 mL). Ethanolamine (61 μL, 1.02 mmol) and N,N-diisopropylethylamine (106 μL, 0.61 mmol) were added sequentially with stirring. The reaction was allowed to proceed to completion at this temperature with continued stirring for 2.5 hours. The reaction was quenched by adding water (6 mL), and the mixture was extracted with dichloromethane (10 mL × 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give the brownish-red title compound (56 mg).

[0259] LC-MS: 767.3 [M+H] + ;

[0260] 1 H NMR (400MHz, Chloroform-d): δ14.04(s,1H),13.27(s,1H),8.04(dd,J=7.8,1.1Hz,1H),7.79(dd,J=8.4,7.7Hz,1H ),7.60(t,J=5.8Hz,1H),7.39(dd,J=8.6,1.1Hz,1H),6.47(s,2H),5.70–5.62(m,1H),5.36(dd,J=4.0,2.1Hz,1H), 5.01(s,1H),4.08(s,3H),4.02–3.93(m,1H),3.88–3.75(m,13H),3.67–3.39(m,3H),3.25(d,J=2.4Hz,2H),2.64(s ,1H),2.44(dd,J=15.0,4.0Hz,1H),2.32(d,J=15.0Hz,1H),2.22(s,3H),2.02–1.85(m,2H),1.38(d,J=6.5Hz,3H).

[0261] Step 3:

[0262] tert-Butylmethyl (2-((2S,4S)-2,5,12-trihydroxy-4-(((2R,4S,5S,6S)-5-hydroxy-6-methyl-4-(methyl(3,4,5-trimethoxybenzyl)amino)tetrahydro-2H-pyran-2-yl)oxy)-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetrabenzo-2-carbamate)ethyl)carbamate (compound 10-2)

[0263] At room temperature, compound 10-1 (144.6 mg, 0.2 mmol) and HATU (155.9 mg, 0.41 mmol) were dissolved in a mixed solvent of DMF and dichloromethane (1:4, 5 mL). 2-(N-Boc-N-methylamino)ethylamine (176 mg, 1.02 mmol) and N,N-diisopropylethylamine (106 μL, 0.61 mmol) were added sequentially with stirring. The reaction was allowed to proceed to completion at this temperature with continued stirring for 2.5 hours. The reaction was quenched by adding water (6 mL), and the mixture was extracted with dichloromethane (10 mL × 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give the reddish-brown title compound (72 mg).

[0264] Step Four:

[0265] Synthesis of compound A-51

[0266] At room temperature, zinc bromide (277.0 mg, 1.23 mmol) was added to a solution of compound 10⁻² (72 mg, 0.082 mmol) in dichloromethane (5 mL), and the reaction was carried out at room temperature for 3 hours, monitored by LC-MS. 5 mL of water was added, and the mixture was stirred for 5 minutes. Then, 5 mL of saturated sodium bicarbonate solution was added, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give the brownish-red title compound (23 mg).

[0267] LC-MS: 780.3 [M+H] + ;

[0268] 1H NMR (400MHz, Chloroform-d): δ14.03(s,1H),13.27(s,1H),8.03(dd,J=7.8,1.1Hz,1H),7.77(t,J=8.1Hz,1H),7.61(t,J=5.8Hz,1H),7.3 6(dd,J=8.6,1.1Hz,1H),6.44(s,2H),5.68–5.58(m,1H),5.33(dd,J=4.0,2.2Hz,1H),5.06(d,J=48.4Hz,1H),4.14–3.96(m,4H),3.81(d, J=6.4Hz,10H),3.62(d,J=13.3Hz,1H),3.58–3.46(m,1H),3.40(d,J=13.3Hz,1H),3.25(d,J=1.8Hz,2H),2.97–2.81(m,2H),2.60(dd,J=9 .5,6.0Hz,1H),2.53(s,3H),2.42(dd,J=14.9,3.9Hz,1H),2.34(d,J=14.9Hz,1H),2.16(s,3H),2.01–1.83(m,2H),1.39(d,J=6.6Hz,3H).

[0269] Using a similar method as described above, the following compounds were prepared.

[0270] Table 1

[0271] Example 8

[0272] Synthesis of (8S,10S)-6,8,11-trihydroxy-10-(((2R,4S,5S,6S)-5-hydroxy-6-methyl-4-(piperazin-1-yl)tetrahydro-2H-pyran-2-yl)oxy)-8-(2-hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione (compound B-1)

[0273] Step 1:

[0274] Synthesis of tert-butyl di(2-carbonylethyl)carbamate (compound 7-2)

[0275] Under nitrogen protection in an ice bath, sodium periodate (3.24 g, 15.2 mmol) was added to a solution of rel-(3R,4S)-3,4-dihydroxypyrrolidine-1-carboxylic acid tert-butyl ester (2.05 g, 10.1 mmol) in THF (15 mL) and water (6 mL). After the addition was complete, the temperature was raised to 20 °C and stirred for 12 hours. The reaction was monitored by LC-MS. The reaction solution was extracted with saturated sodium chloride aqueous solution and ethyl acetate (20 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a colorless oily title compound (2.0 g), which was immediately used directly in the next reaction.

[0276] Step Two:

[0277] Synthesis of tert-butyl 4-((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetrafluorene-1-yl)oxy)tetrahydro-2H-pyran-4-yl)piperazine-1-carboxylic acid ester (compound 7-3)

[0278] Doxorubicin hydrochloride (174 mg, 0.3 mmol) and compound 7-2 (181.1 mg, 0.9 mmol) were mixed in 6 mL of dry DCM and stirred at room temperature for 10 min. The reaction mixture was cooled to 0 °C, and NaBH(OAc)3 (318 mg, 1.5 mmol) was added. The reaction mixture was brought to room temperature over 1 hour with stirring, and the reaction was monitored by LC-MS. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10 mL × 3). The combined organic phases were washed successively with water, 5% sodium bicarbonate solution, and brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the residue, which was purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give the brownish-red title compound (125 mg).

[0279] Step 3:

[0280] Synthesis of compound B-1

[0281] At room temperature, zinc bromide (168.9 mg, 0.75 mmol) was added to a 5 mL solution of compound 7-3 (106.8 mg, 0.15 mmol) in dichloromethane. The reaction was carried out at room temperature for 24 hours, and the reaction was monitored by LC-MS. 6 mL of water was added to the system, and the mixture was stirred for 5 minutes. Then, 5 mL of saturated sodium bicarbonate solution was added, and the mixture was extracted with chloroform (15 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by preparative HPLC (acetonitrile-0.05% ammonium bicarbonate aqueous solution system) to give the brownish-red title compound (66 mg).

[0282] LC-MS: 613.2 [M+H] + ;

[0283] 1 H-NMR (400MHz, DMSO-d6): δ7.96–7.90(m,2H),7.71–7.64(m,1H),5.40(s,1H),5.28(s,1H),4.96(s,1H),4.84(t,J=5.9Hz,1H),4.55(d,J=5. 9Hz,2H),4.00(s,3H),3.95(s,1H),3.60(s,1H),2.98(s,2H),2.40(s,10H),2.15(d,J=6.2Hz,1H),1.25–1.17(m,3H),1.14(d,J=6.4Hz,3H).

[0284] Using a similar method as described above, the following compounds were prepared.

[0285] Table 2

[0286] Example 9

[0287] Synthesis of 1-((2S,3S,4S,6R)-6-(((1S,3S)-3-acetyl-3,5,12-trihydroxy-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)-3-hydroxy-2-methyltetrahydro-2H-pyran-4-yl)piperazine-2-nitrile (compound C-2)

[0288] Step 1:

[0289] Synthesis of (9H-fluorene-9-yl)methyl(2-chloroethyl)(2-hydroxyethyl)carbamate (compound 8-2)

[0290] Compound 8-1 (3.0 g, 18.75 mmol) was dissolved in 1,4-dioxane / water (30 mL, v / v = 1:1), followed by the addition of NaHCO3 (6.3 g, 75 mmol). The reaction was carried out at room temperature for 10 minutes. Then, Fmoc-Cl (4.85 g, 18.75 mmol) was added to the reaction system, and the reaction was carried out at room temperature for 30 minutes. After the starting material reacted completely, the reaction was quenched with water, extracted with dichloromethane (50 mL × 3), and the organic phases were combined. The organic phase was then washed once with saturated brine, collected, dried over anhydrous sodium sulfate, concentrated, and separated by silica gel column chromatography (petroleum ether / ethyl acetate, 2:1). The final product was 2.46 g of the oil, which is the title compound.

[0291] Step Two:

[0292] Synthesis of (9H-fluorene-9-yl)methyl(2-iodoethyl)(2-hydroxyethyl)carbamate (compound 8-3)

[0293] Compound 8-2 (2.46 g, 7.11 mmol) was dissolved in acetone (50 mL), followed by the addition of NaHCO3 (0.18 g, 2.13 mmol) and NaI (5.33 g, 35.57 mmol). The resulting reaction mixture was refluxed at 85 °C for 72 hours, and the reaction was monitored by LC-MS. The mixture was cooled to room temperature, quenched with water, and extracted with dichloromethane (50 mL × 2). The organic phases were combined, washed once with saturated brine, and then combined again. The organic phases were dried over anhydrous sodium sulfate, concentrated, and separated by reverse-phase reaction (water / acetonitrile, 45:55). The concentrate yielded 1.4 g of the oily substance, which is the title compound.

[0294] Step 3:

[0295] Synthesis of (9H-fluorene-9-yl)methyl(2-iodoethyl)(2-oxoethyl)carbamate (compound 8-4)

[0296] Compound 8-3 (300 mg, 0.69 mmol) was dissolved in dichloromethane (15 mL). The reaction mixture was then cooled to 5 °C in an ice-water bath, purged with nitrogen three times, and then Dess-Martin periodinane (873 mg, 2.06 mmol) was added. The mixture was stirred for 5 minutes and then allowed to react at room temperature for 2 hours. The reaction mixture was filtered, and the filter cake was washed with methyl tert-butyl ether. The filtrates were combined, concentrated, and the residue was purified by rapid silica gel column chromatography (petroleum ether / ethyl acetate, 3:1) to give 186 mg of a colorless oil, which is the title compound.

[0297] Step Four:

[0298] Synthesis of (9H-fluorene-9-yl)methyl 4-((2S,3S,4S,6R)-6-(((1S,3S)-3-acetyl-3,5,12-trihydroxy-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)-3-hydroxy-2-methyltetrahydro-2H-pyran-4-yl)-3-cyanopiperazine-1-carboxylic acid ester (compound 8-5)

[0299] Add daunorubicin hydrochloride (50 mg, 0.089 mmol), potassium cyanide (8.08 mg, 0.124 mmol), and methanol (3 mL) to a 10 mL round-bottom flask. Stir at room temperature for 10 minutes, then add a methanol solution (1 mL) of compound 8-4 (50 mg, 0.115 mmol). Continue stirring the reaction mixture at room temperature for 16 hours. Pour the reaction solution into water (10 mL), extract with ethyl acetate (15 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure to dryness, and purify the residue by preparative TLC (dichloromethane / methanol, 15:1) to obtain 30.0 mg of crude orange-red product.

[0300] Step 5:

[0301] Synthesis of 1-((2S,3S,4S,6R)-6-(((1S,3S)-3-acetyl-3,5,12-trihydroxy-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)-3-hydroxy-2-methyltetrahydro-2H-pyran-4-yl)piperazine-2-nitrile (compounds 8-6)

[0302] Compound 8-5 (30 mg, 0.036 mmol) was dissolved in DMF (3 mL), and diethylamine (38 μL, 0.36 mmol) was added at room temperature. The mixture was stirred for 2 hours, and the reaction was monitored by LC-MS. The reaction solution was concentrated under reduced pressure, and the residue was purified by rapid silica gel column chromatography (dichloromethane / methanol, 10:1) to obtain 11 mg of the brownish-red product, which was immediately used in the next reaction.

[0303] Step Six:

[0304] Synthesis of compound C-2

[0305] Acetyl chloride (3 μL, 0.036 mmol) was added to a mixture of compound 8-6 (11 mg, 0.018 mmol) and triethylamine (7 μL, 0.048 mmol) in THF (0.5 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. After concentration, the residue was purified by preparative HPLC (acetonitrile-0.1% ammonium bicarbonate aqueous solution) to give 5 mg of a brownish-red solid product.

[0306] LC-MS: 664.2 [M+H] + ;

[0307] 1H NMR (400MHz, CDCl3): δ14.04(d,J=3.4Hz,1H),13.30(s,1H),8.05(d,J=7.5Hz,1H),7.81–7.74(m,1H),7.42–7.35(m,1H),5 .64–5.54(m,1H),5.39–5.23(m,2H),4.74–4.67(m,1H),4.45–4.25(m,1H),4.09(d,J=1.9Hz,3H),4.03–3.88(m,1H),3.74(d ,J=34.7Hz,2H),3.41(d,J=13.3Hz,1H),3.23(t,J=19.2Hz,2H),3.09–2.92(m,1H),2.85(d,J=12.8Hz,1H),2.72(d,J=11.1H z,1H),2.58–2.51(m,1H),2.42(d,J=3.1Hz,3H),2.38–2.19(m,3H),2.14–2.07(m,4H),2.04–1.90(m,2H),1.37–1.3(m,2H).

[0308] Example 10

[0309] Synthesis of (2S,4S)-4-(((2R,4S,5S,6S)-4-(3-cyanomorpholine)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-N-(2-(methylamino)ethyl)-6,11-dioxo-1,2,3,4,6,11-hexahydrotetrabenzo-2-carboxamide (compound C-10)

[0310] Synthesis of (2S,4S)-4-(((2R,4S,5S,6S)-4-(3-cyanomorpholine)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-N-(2-hydroxyethyl)-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetrabenzo-2-carboxamide (compound C-12)

[0311] Step 1:

[0312] Synthesis of 4-((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetraphenyl-1-yl)oxy)tetrahydro-2H-pyran-4-yl)morpholine-3-nitrile (compound 9-1)

[0313] Compound 1-1 (80 mg, 0.138 mmol), KCN (16 mg, 0.248 mmol), and methanol (5 mL) were added to a 10 mL round-bottom flask. The mixture was stirred at room temperature for 10 min, followed by the addition of a tetrahydrofuran solution (1.5 mL) of compound 5-3 (99 mg, 0.42 mmol). The mixture was stirred at 25 °C for 40 h. The reaction mixture was then added to water and extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure. The concentrate was purified by preparative TLC (dichloromethane / methanol, 20:1) to give 76 mg of a brownish-red solid, which was the title compound.

[0314] Step Two:

[0315] Synthesis of (2S,4S)-4-(((2R,4S,5S,6S)-4-(3-cyanomorpholine)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetrabenzo-2-carboxylic acid (compound 9-2)

[0316] At room temperature, compound 9-1 (76 mg, 0.1 mmol) was dissolved in a methanol / water mixture (3:2, 20 mL). Then, while stirring, an aqueous solution of sodium periodate (32.6 mg, 0.152 mmol) (3 mL) was added dropwise to the solution. The reaction was continued at room temperature for 30 min until complete. The reaction solution was extracted with dichloromethane (15 mL × 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was used directly as the crude product in the next reaction step.

[0317] Step 3:

[0318] Synthesis of tert-butyl(2-((2S,4S)-4-(((2R,4S,5S,6S)-4-(3-cyanomorpholine)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrotetrabenzo-2-carbamate)ethyl)(methyl)carbamate (compound 9-3)

[0319] At room temperature, compound 9-2 (31 mg, 0.05 mmol) and HATU (39 mg, 0.1 mmol) were dissolved in a mixed solvent of DMF and dichloromethane (1:4, 2 mL). 2-(N-Boc-N-methylamino)ethylamine (44 mg, 0.25 mmol) and N,N-diisopropylethylamine (27 μL, 0.15 mmol) were added sequentially with stirring. The reaction was allowed to proceed to completion at this temperature with continued stirring for 2.5 hours. The reaction was quenched by adding water (2 mL), and the mixture was extracted with dichloromethane (10 mL × 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give the reddish-brown title compound (32 mg).

[0320] Step Four:

[0321] Synthesis of compound C-10

[0322] At room temperature, zinc bromide (158.8 mg, 0.705 mmol) was added to a solution of compound 9-3 (32 mg, 0.047 mmol) in dichloromethane (3 mL), and the reaction was carried out at room temperature for 3 hours, monitored by LC-MS. 3 mL of water was added, and the mixture was stirred for 5 minutes. Then, 3 mL of saturated sodium bicarbonate solution was added, and the mixture was extracted with dichloromethane (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give the brownish-red title compound (12 mg).

[0323] LC-MS: 681.3 [M+H] + ;

[0324] 1H-NMR (400MHz, CDCl3): δ13.99(s,1H),7.98(d,J=7.7Hz,1H),7.72(td,J=8.1,3.7Hz,1H),7.45(s,1H),7.32(d,J=8.5Hz,1H), 5.55(d,J=7.9Hz,1H),5.25(d,J=19.6Hz,1H),4.02(s,3H),4.00–3.91(m,2H),3.90–3.77(m,2H),3.69-3.62(m,2H),3.56–3.47( m,1H),3.44–3.30(m,2H),3.19(s,2H),2.91–2.79(m,1H),2.75(d,J=5.8Hz,1H),2.61(d,J=12.0Hz,1H),2.42(s,3H),2.36(d,J =4.9Hz,1H),2.29–2.20(m,2H),1.94(d,J=5.5Hz,1H),1.84-1.80(m,1H),1.77–1.69(m,2H),1.39(s,2H),1.28(t,J=6.3Hz,3H).

[0325] Step 5:

[0326] Synthesis of compound C-12

[0327] At room temperature, compound 9-2 (30 mg, 0.05 mmol) and HATU (39 mg, 0.1 mmol) were dissolved in a mixed solvent of DMF and dichloromethane (1:4, 2 mL). Ethanolamine (15 μL, 0.25 mmol) and N,N-diisopropylethylamine (27 μL, 0.15 mmol) were added sequentially with stirring. The reaction was allowed to proceed to completion by stirring at this temperature for 2.5 hours. The reaction was quenched by adding water (2 mL), and the mixture was extracted with dichloromethane (10 mL × 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by rapid silica gel column chromatography (dichloromethane / methanol, 95:5) to give the brownish-red title compound (18 mg).

[0328] LC-MS: 668.2 [M+H] + ;

[0329] 1H-NMR (400MHz, CDCl3): δ14.08(s,1H),13.28(s,1H),8.05(dd,J=7.4,2.0Hz,1H),7.79(td,J=8.1,3.7Hz,1H),7.63(d,J= 5.5Hz,1H),7.40(dd,J=8.1,2.8Hz,1H),5.67–5.61(m,1H),5.36(dd,J=11.4,6.6Hz,2H),4.82(d,J=25.4Hz,1H),4.03–4.01 (m,2H),3.97–3.88(m,2H),3.79(s,J=16.1Hz,3H),3.73–3.69(m,3H),3.56(d,J=15.2Hz,3H),3.26(s,1H),2.79–2.53(m,4H ),2.50–2.41(m,1H),2.36–2.28(m,2H),2.23-2.17(m,1H),2.01(d,J=5.7Hz,1H),1.81-1.76(m,1H),1.36(t,J=6.3Hz,3H).

[0330] Cell killing assay of anthracycline compounds

[0331] The purpose of this experiment was to detect the in vitro cell proliferation inhibitory activity of the representative compound of this application and the control load molecule Dxd on MDA-MB-231 (human breast cancer cells, Nanjing Kebai), NCI-H322 (human lung cancer cells, Nanjing Kebai), and HCT-116 (human colorectal cancer cells, Nanjing Kebai).

[0332] Experimental equipment:

[0333] 96-hole transparent flat-bottom whiteboard (Corning, #3610)

[0334] RPMI 1640 medium (Gibco, #A10491-01)

[0335] DMEM medium (Gibco, #11995065)

[0336] Fetal Bovine Serum (Sigma-Aldrich, #F8687-500ML)

[0337] Luminescent Cell Viability Assay (Promega, #G7572)

[0338] Experimental methods:

[0339] Human breast cancer cells MDA-MB-231, human lung cancer cells NCI-H322, and human colorectal cancer cells HCT-116 were washed with PBS, digested with 0.25% Trypsin-EDTA for about 3-10 minutes, digestion was stopped with complete culture medium, the cells were centrifuged at 1000 rpm for 3 minutes, the supernatant was discarded, the cells were resuspended in complete cell culture medium, the cells were counted with a cell counter, the cells were adjusted to the required density, 90.0 μL of cell suspension (3000 cells / well) was added to each well, and the cell plate was incubated overnight in a 37°C, 5% CO2 cell culture incubator.

[0340] In the experiment, a DMSO control group and a test sample group were set up. The cell plates were removed from the incubator to observe the cell adhesion status. After the cells adhered, 10.00 μL of sample (starting at a final concentration of 10 μM, 5-fold dilution, 9 concentrations) was added to each well, gently shaken, and then incubated in a 37℃, 5% CO2 incubator.

[0341] After 3 days of incubation, add 100.0 μL / well CellTiter-Glo TM (Promega, catalog number: G7572) working solution, shaken in a constant temperature shaker to lyse cells, and read the plate on a microplate reader after 10 minutes.

[0342] Formula for calculating cell proliferation inhibition rate: Cell proliferation inhibition rate % = (1 - RLU) 供试品孔 / RLU DMSO对照孔 )×100%.

[0343] Data analysis: A graph was plotted with the logarithmic value of the sample concentration on the x-axis and the inhibition percentage on the y-axis. Nonlinear regression (curve fit) analysis was performed on the data to obtain the IC50 of each test sample. 50 value.

[0344] Table 3. In vitro cell proliferation inhibition activity

[0345] Conclusion: As shown in Table 3, the compounds of this application have in vitro cell proliferation inhibitory activity.

Claims

1. The compound represented by formula (I), its pharmaceutically acceptable salt, and its isomers: in, R1 is selected from H and C. 1-6 Alkoxy; R2 is selected from O and NH; R3 is selected from C 1-6 Alkyl, hydroxy C 1-6 Alkyl, -C(O)C 1-6 Alkyl, -C(O)NR n1 R n2 Wherein C 1-6 Alkyl groups may be optionally replaced by hydroxyl groups or -OC(O)CH2NH2; R n1 and R n2 Each is independently selected from H and C. 1-6 Alkyl, 3- to 8-membered cycloalkyl, 6- to 10-membered aryl; wherein, the C 1-6 Alkyl, 3- to 8-membered cycloalkyl, 6- to 10-membered aryl, optionally with hydroxyl, hydroxyl C 1-6 The alkyl group is substituted with one or more substituents selected from alkyl, amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, 5- to 10-membered heteroaryl, and 6- to 10-membered aryl, wherein the amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, 5- to 10-membered heteroaryl, and 6- to 10-membered aryl are optionally selected from C10. 1-6 Alkyl, amino, hydroxy C 1-6 One or more substituents in the alkyl group are substituted; or, R n1 and R n2 The atoms bonded to it together form a 3- to 8-membered heterocyclic group, wherein the 3- to 8-membered heterocyclic group is optionally bonded to C 1-6 Alkyl substitution, wherein the C 1-6 Alkyl groups may be optionally substituted with amino or hydroxyl groups; R4 and R5 are each independently selected from H and C. 1-6 Alkyl, -C(O)C 1-6 Alkyl, 3- to 8-membered cycloalkyl, 3- to 8-membered heterocyclic, wherein the C 1-6 The alkyl group is optionally substituted with a 6- to 10-membered aryl group, a 4- to 10-membered bridged cycloyl group, a 3- to 8-membered cycloalkyl group, a 5- to 10-membered heteroaryl group, and a 3- to 8-membered heterocyclic group; wherein the 6- to 10-membered aryl group, the 4- to 10-membered bridged cycloyl group, and the 5- to 10-membered heteroaryl group are optionally selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, halogenated C 1-6 The substance is substituted by one or more substituents selected from alkyl, cyano, 3- to 8-membered cycloalkyl, -NO2, and amino groups; Alternatively, R4 and R5, together with the nitrogen atom they are attached to, can form a 3- to 8-membered heterocyclic group or a 3- to 8-membered heteroaryl group, wherein the 3- to 8-membered heterocyclic group or the 3- to 8-membered heteroaryl group is optionally bonded by CN or C. 1-6 Alkoxy, C 1-6 Alkyl, -C(O)OC 1-6 Alkyl, -C(O)C 3-8 Cycloalkyl, -C(O)NR n3 R n4 -C(O)C 1-6 One or more substituents in the alkyl group are replaced, wherein the C 1-6 Alkyl groups are optionally surrounded by 3 to 8-membered heterocyclic groups, C 1-6 Alkoxy, NR n3 R n4 It is substituted by one or more substituents among 5- to 10-membered heteroaryl, amino, and hydroxyl groups; R n3 and R n4 Each is independently selected from H and C. 1-6 Alkyl, -C(O)C 1-6 alkyl; Furthermore, the compound of formula I is not any of the following compounds:

2. The compound represented by formula (I) according to claim 1, its pharmaceutically acceptable salt, its isomer, selected from formula (IA): Wherein R1, R2, R3, R4, and R5 are as defined in claim 1, and the compound of formula (IA) is not any of the following compounds:

3. The compound according to claim 1 or 2, its pharmaceutically acceptable salt, its isomer, wherein R1 is selected from -OCH3 or H, preferably -OCH3.

4. The compound according to any one of claims 1-3, its pharmaceutically acceptable salt, its isomer, wherein R2 is O.

5. The compound according to any one of claims 1-4, its pharmaceutically acceptable salt, its isomer, wherein R3 is selected from -C(O)C 1-6 Alkyl and -C(O)NR n1 R n2 Wherein C 1-6 Alkyl groups may be optionally replaced by hydroxyl groups and -OC(O)CH2NH2; R n1 H is independent; R n2 Selected independently from C 1-6 Alkyl and 6-10 aryl groups; the C 1-6 The alkyl and 6-10 aryl groups are optionally substituted with one or more substituents selected from hydroxyl, amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, 5- to 10-membered heteroaryl, and 6-10 aryl groups; wherein the amino, 3- to 8-membered cycloalkyl, 4- to 10-membered bridged cycloyl, 3- to 8-membered heterocyclic, and 6-10 aryl groups are optionally selected from C 1-6 Replaced by one or more substituents in alkyl and amino groups; Or, R n1 and R n2 The atoms bonded to it together form a 3- to 8-membered heterocyclic group, wherein the 3- to 8-membered heterocyclic group is optionally bonded to C 1-6 Alkyl substitution, wherein, The C mentioned 1-6 Alkyl groups may be optionally substituted with amino and hydroxyl groups; Preferred, R3 is selected from -C(O)C 1-6 Alkyl and -C(O)NR n1 R n2 Wherein C 1-6 Alkyl groups may be optionally substituted with hydroxyl groups; R n2 Selected independently from C 1-6 Alkyl and 6-10 aryl groups; the C 1-6 The alkyl and 6-10 aryl groups are optionally substituted with one or more substituents selected from hydroxyl, amino, and 6-10 aryl groups; wherein the amino and 6-10 aryl groups are optionally selected from C 1-6 Replaced by one or more substituents in alkyl and amino groups; More preferably, R3 is selected from 6. The compound according to any one of claims 1-5, its pharmaceutically acceptable salt, its isomer, wherein, R4 is independently selected from C 1-6 Alkyl and -C(O)C 1-6 alkyl; R5 is independently selected from C 1-6 Alkyl, 3- to 8-membered cycloalkyl, and 3- to 8-membered heterocyclic groups; wherein, the C 1-6 The alkyl group is optionally substituted with a 6- to 10-membered aryl group, a 4- to 10-membered bridged cycloyl group, a 3- to 8-membered cycloalkyl group, a 5- to 10-membered heteroaryl group, and a 3- to 8-membered heterocyclic group; said 6- to 10-membered aryl group, 4- to 10-membered bridged cycloyl group, and 5- to 10-membered heteroaryl group are optionally selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, halogenated C 1-6 Substituted by one or more substituents from alkyl, cyano, 3- to 8-membered cycloalkyl, -NO2, and amino groups; R n3 and R n4 Each is independently selected from H and C. 1-6 Alkyl and -C(O)C 1-6 alkyl; Alternatively, R4, R5, and the nitrogen atom to which they are attached can form a 3- to 8-membered heterocyclic group; wherein the 3- to 8-membered heterocyclic group is optionally bonded by CN, C 1-6 Alkyl, -C(O)C 3-8 Cycloalkyl, -C(O)NR n3 R n4 -C(O)C 1-6 One or more substituents in the alkyl group are substituted; the C 1-6 Alkyl groups are optionally C 1-6 It is substituted by one or more substituents among alkoxy, amino, and hydroxyl groups; R n3 and R n4 Each is independently selected from H and C. 1-6 alkyl; Preferred, R4 is independently C 1-6 alkyl; R5 is independently selected from C 1-6 Alkyl groups and 3 to 8-membered heterocyclic groups, wherein the C 1-6 The alkyl group is optionally substituted with a 6- to 10-membered aryl group, a 4- to 10-membered bridged cycloyl group, a 5- to 10-membered heteroaryl group, and a 3- to 8-membered heterocyclic group; wherein the 6- to 10-membered aryl group, the 4- to 10-membered bridged cycloyl group, and the 5- to 10-membered heteroaryl group are optionally selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, halogenated C 1-6 Substituted by one or more substituents from alkyl, cyano, 3- to 8-membered cycloalkyl, -NO2, and amino groups; R n3 and R n4 Each is independently selected from H and C. 1-6 alkyl; Alternatively, R4, R5, and the nitrogen atom to which they are attached can form a 3- to 8-membered heterocyclic group; the 3- to 8-membered heterocyclic group may optionally be coated with CN, -C(O)C 3-8 Cycloalkyl, -C(O)C 1-6 One or more substituents in the alkyl group are substituted; More preferably, R5 is independently C 1-6 Alkyl; wherein, the C 1-6 The alkyl group is optionally substituted with a 6- to 10-membered aryl group and a 5- to 10-membered heteroaryl group; wherein the 6- to 10-membered aryl group and the 5- to 10-membered heteroaryl group are optionally selected from C 1-6 Alkoxy, C 1-6 Alkyl, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, halogenated C 1-6 Substituted with one or more of the following substituents: alkyl, cyano, 3- to 8-membered cycloalkyl, -NO2, and amino; R n3 and R n4 Each is independently selected from H and C. 1-6 alkyl; For example, R4 is independently CH3; R5 is independently C 1-6 Alkyl; wherein, the C 1-6 The alkyl group may optionally be substituted with phenyl, pyrimidinyl, or pyridinyl groups; wherein the phenyl, pyrimidinyl, or pyridinyl groups are C-substituted. 1-6 Alkoxy, -NR n3 R n4 -CH2-NR n3 R n4 3 to 8-membered heterocyclic groups, halogens, cyano groups, 3 to 8-membered cycloalkyl groups, and -NO2 substitutions; For example, -NR4R5 is selected from the following structure:

7. The compound represented by formula (II), its pharmaceutically acceptable salt, and its isomers: in, R4 is selected from H and C. 1-6 Alkyl groups, preferably CH3; Ring A is selected from C6-C 10 Aryl, preferably phenyl; R 5-1 Selected from C 1-6 Alkoxy, C 1-6 Alkyl group, preferably -OCH3; p is selected from integers of 0, 1, 2, 3.

8. The compound represented by formula (III), its pharmaceutically acceptable salt, and its isomers: in, R1 is C 1-6 alkoxy group; R3 is -C(O)C 1-6 Alkyl, the C 1-6 Alkyl groups are replaced by hydroxyl groups; Alternatively, R1 can be H and R3 can be -C(O)C. 1-6 alkyl; Preferred, R1 is C 1-6 alkoxy group; R3 is -C(O)CH2OH; Alternatively, R1 can be H and R3 can be -C(O)CH3.

9. The compound represented by formula (IV-1), its pharmaceutically acceptable salt, and its isomers: in, R 5-6 Selected from C 1-6 Alkyl groups and -C(=O)-R 5-2 The C 1-6 Alkyl groups are optionally C 1-6 Alkyl substitution; R 5-2 Selected from C 1-6 Alkyl, -NH-C 1-6 Alkyl, 3- to 8-membered cycloalkyl, wherein the C 1-6 Alkyl groups may be optionally replaced by hydroxyl or amino groups; R 5-3 Selected from C 1-6 Alkyl, the C 1-6 Alkyl groups may be optionally substituted with hydroxyl groups; Preferred, R 5-6 for R 5-3 Selected from CH3 and CH2OH; The optimal choice, R 5-6 for 10. The compound represented by formula (IV-2), its pharmaceutically acceptable salt, and its isomers: in, R n1 and R n2 Each is independently selected from H and C. 1-6 Alkyl, the C 1-6 Alkyl groups may be optionally amino- or C-shaped. 1-6 The alkylamino group, hydroxyl group, 4- to 10-membered bridged cycloalkyl group, 3- to 8-membered cycloalkyl group, 3- to 8-membered heterocyclic group, and 5- to 10-membered heteroaryl group are substituted, wherein the 4- to 10-membered bridged cycloalkyl group, 3- to 8-membered cycloalkyl group, 3- to 8-membered heterocyclic group, and 5- to 10-membered heteroaryl group are optionally substituted with amino groups; or, R n1 R n2 Together with the nitrogen atoms they are attached to, they form 3- to 8-membered heterocyclic groups, which are optionally C-shaped. 1-6 Alkyl substitution, wherein C 1-6 Alkyl groups may be optionally replaced by amino or hydroxyl groups.

11. A compound, its pharmaceutically acceptable salt, and its isomers, wherein the compound is selected from:

12. An antibody-drug conjugate comprising a small molecule drug, a linker, and an antibody, wherein the small molecule drug is selected from the compounds of any one of claims 1-11, pharmaceutically acceptable salts thereof, or isomers thereof.

13. A pharmaceutical composition comprising a compound according to any one of claims 1-11, a pharmaceutically acceptable salt thereof, an isomer thereof, and one or more pharmaceutically acceptable carriers.

14. Use of the compound, pharmaceutically acceptable salt thereof, isomer thereof, or antibody-drug conjugate according to any one of claims 1-11, or pharmaceutical composition according to claim 13, in the preparation of a medicament for the treatment and / or prevention of tumors.

15. A method of treating and / or preventing tumors, comprising administering to an individual in need an effective amount of the compound of any one of claims 1-11, a pharmaceutically acceptable salt thereof, an isomer thereof, or an effective amount of the antibody-drug conjugate of claim 12, or an effective amount of the pharmaceutical composition of claim 13.

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