Deuterated camptothecin compounds, methods for producing the same, and their use

Abstract

This invention discloses a deuterated camptothecin compound represented by formula II and a pharmaceutically acceptable salt thereof, a method for producing the same, a pharmaceutical composition comprising the deuterated camptothecin compound or a pharmaceutically acceptable salt thereof, and the use thereof in the treatment of tumor-related diseases. This deuterated camptothecin compound has excellent pharmacoactivity and pharmacokinetic properties, reduced toxicity and improved pharmacosafety. [Formula 1] JPEG2026516064000069.jpg51169

Description

Detailed description of the invention

[0001] [Cross-citation of related applications] This application claims the rights and priority of Patent Application No. 202310506357.6 filed with the State Intellectual Property Administration of China on May 6, 2023, Patent Application No. 202311533730.3 filed with the State Intellectual Property Administration of China on November 16, 2023, and Patent Application No. 202410535444.9 filed with the State Intellectual Property Administration of China on April 29, 2024, the entire contents of which are incorporated herein by reference. [Technical Field] This disclosure pertains to the field of medicinal chemistry and, more specifically, to deuterated camptothecin compounds and pharmaceutically acceptable salts thereof, methods for producing the same, pharmaceutical compositions comprising the deuterated camptothecin compounds or pharmaceutically acceptable salts thereof, and the use of the deuterated camptothecin compounds or pharmaceutically acceptable salts thereof or the pharmaceutical compositions in the treatment of tumor-related diseases. [Background technology] Camptothecin (CPT) is a pyrroloquinoline cytotoxic alkaloid and one of the most researched natural antitumor drugs after paclitaxel. It is mainly found in the fruit or root bark of Camptotheca acuminata, a plant of the Cornaceae family endemic to China. In 1985, Hsiang et al. discovered the mechanism by which camptothecin and its derivatives exert their anticancer effects by inhibiting DNA synthesis by targeting topoisomerase I (Topo I). This discovery has since attracted widespread attention, leading to the development of numerous derivatives and making it a new research focus in the field of anticancer. Subsequently, new-generation camptothecin-based drugs such as 10-hydroxycamptothecin (HCPT), irinotecan, topotecan, SN-38, and berotecan were successively approved and launched, and are used to treat tumor diseases such as colorectal cancer, small cell lung cancer, and ovarian cancer. Research on their indications and dosage forms has also yielded remarkable results.

[0002] With the development of drug delivery systems, a series of camptothecin derivatives that could not be used as drugs in the past and had significant side effects have been reused. However, the problem of the in vivo toxicity of camptothecin-based drugs has still not been completely solved. How to further reduce the toxicity of these drugs remains a topic of current concern. 〔Summary of the Invention〕 In one aspect, the present disclosure provides a compound of formula II or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

[0003]

Chemical Formula

[0004] However, R1 is selected from H, halogen, OH, SH, NH2, C 1-4 , 1-4 , 1-4 , alkyl, C 1-4 haloalkyl or C 1-4 alkoxy, R2 is selected from H, halogen, C 1-4 alkyl or C 1-4 alkoxy, Alternatively, R1 and R2 cyclize to -O-(CH2) m -O-, where m is selected from 1, 2, 3, R3 and R4 are each independently selected from H, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 haloalkyl or C 1-4 haloalkoxy, Alternatively, R3 and R4 cyclize to -(CH2) k -, where k is selected from 1, 2, 3, 4, X is selected from H, OH, HO-CH(R5)-(CH2) p -CO-NH- or -N(R6)(R7), where p is selected from 0, 1, 2, R5 is selected from H, C 1-4 alkyl, C 1-4 haloalkyl, 3-6 member cycloalkyl or 3-6 member heterocycloalkyl, R6 is selected from H, C 1-4Alkyl or C 1-4 Selected from haloalkyls, R7 is selected from H or R8-S(O)2-. R8 is C 1-4 Selected from alkyl, and t is selected from 0, 1, 2, 3, 4, and 5.

[0005] In this disclosure, "R1 and R2 are -O-(CH2) m The expression "to form an O-" means that R1 and R2 are linked together to form an O-(CH2) m This refers to the formation of -O-. k The expression "to form a ring" means that R3 and R4 are linked together to form (CH2) k - refers to forming something.

[0006] In any embodiment of the compound of formula II in this disclosure, R1 is C 1-4 It is an alkyl group, and R2 is a halogen.

[0007] In any embodiment of the compound of formula II of this disclosure, R1 is methyl and R2 is F.

[0008] In any embodiment of the compound of formula II of this disclosure, R1 is H and R2 is H.

[0009] In any embodiment of the compound of formula II of this disclosure, R1 and R2 are cyclized to -O-CH2-O-.

[0010] In any embodiment of the compound of formula II of this disclosure, R1 is NH2 and R2 is H or a halogen. In any embodiment of the compound of formula II of this disclosure, R1 is NH2 and R2 is H or F.

[0011] In any embodiment of the compound of formula II of this disclosure, R3 is H and R4 is H.

[0012] In any embodiment of the compound of formula II of this disclosure, R3 is H and R4 is C 1-4 It is alkyl.

[0013] In any embodiment of the compound of formula II of this disclosure, R3 is H and R4 is methyl.

[0014] In any embodiment of the compound of formula II of this disclosure, R3 and R4 are cyclized to -CH2-CH2-.

[0015] In any embodiment of the compound of formula II in this disclosure, X is HO-CH(R5)-(CH2) p -CO-NH-, where p is selected from 0, 1, and 2.

[0016] In any embodiment of the compound of formula II of this disclosure, X is selected from OH or NH2.

[0017] In any embodiment of the compound of formula II of this disclosure, X is H and t is 0.

[0018] In any embodiment of the compound of formula II of this disclosure, X is -N(R6)(R7).

[0019] In one embodiment of the compound of formula II of the present disclosure, the compound of formula II is the compound of formula IIa, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0020] [ka]

[0021] However, R1, R2, R3, and R4 are as defined in any one embodiment of the compound of formula II, and X1 is H, HO-CH(R5)-(CH2) p -CO- is selected, p is selected from 0, 1, 2, R5 is H, C 1-4Selected from alkyl, 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl.

[0022] In any embodiment of the compound of formula IIa of this disclosure, R1 is selected from methyl or methoxy, or R1 is methyl.

[0023] In any embodiment of the compound of formula IIa of this disclosure, R2 is selected from F or Cl, or R2 is F.

[0024] In any embodiment of the compound of formula IIa of this disclosure, R1 is methyl and R2 is F.

[0025] In any embodiment of the compound of formula IIa in this disclosure, R3 and R4 are -(CH2) k - is cyclically transformed, where k is 2.

[0026] In any embodiment of the compound of formula IIa of this disclosure, R3 is H and R4 is H.

[0027] In any embodiment of the compound of formula IIa in this disclosure, X1 is HO-CH(R5)-(CH2) p -CO- where p is selected from 0 or 1, and R5 is H, C 1-4 Selected from alkyl or 3-6 membered cycloalkyl groups.

[0028] In any embodiment of the compound of formula IIa in this disclosure, X1 is HO-CH(R5)-(CH2) p It is -CO-, where p is selected from 0 or 1, and R5 is selected from H, methyl, or cyclopropyl.

[0029] In any embodiment of the compound of formula IIa of this disclosure, X1 is HO-CH(R5)-CO- and R5 is selected from H or a 3- to 6-membered cycloalkyl group.

[0030] In any embodiment of the compound of formula IIa of this disclosure, X1 is HO-CH(R5)-CO- and R5 is selected from H or cyclopropyl.

[0031] In any embodiment of the compound of formula IIa of this disclosure, X1 is HO-CH(R5)-CH2-CO- and R5 is C 1-4 It is alkyl.

[0032] In any embodiment of the compound of formula IIa of this disclosure, X1 is HO-CH(R5)-CH2-CO- and R5 is methyl.

[0033] In any embodiment of the compound of formula IIa in this disclosure, X1 is H,

[0034] [ka]

[0035] Selected from.

[0036] In some embodiments of the compounds of formula IIa of this disclosure, the compound of formula IIa is selected from the compounds of formula IIa-1 or formula IIa-2 below, or pharmaceutically acceptable salts thereof, or stereoisomers thereof.

[0037] [ka]

[0038] However, R1, R2, and X1 are as defined in any one embodiment of the compound of formula IIa.

[0039] In some embodiments of the compounds of formula IIa of this disclosure, the compound of formula IIa is selected from the following compounds or pharmaceutically acceptable salts thereof or stereoisomers thereof.

[0040] [ka]

[0041] In one embodiment of the compound of formula II of the present disclosure, the compound of formula II is the compound of formula IIb or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

[0042] [ka]

[0043] However, R1 is H, OH, halogen, NH2, C 1-4 Alkyl, C 1-4 Haloalkyl or C 1-4 Selected from alkoxy, R2 is H, halogen, C 1-4 Alkyl or C 1-4 Selected from alkoxy, Alternatively, R1 and R2 are -O-(CH2) m We cyclize to -O-, where m is selected from 1, 2, and 3. R3 and R4 are independently H and C 1-4 Alkyl or C 1-4 Selected from alkoxy, Alternatively, R3 and R4 are -(CH2) k - is revolved around k, where k is selected from 1, 2, and 3. X2 is selected from H, OH, or -N(R6)(R7), R6 is H or C 1-4 Selected from alkyl groups, R7 is selected from H or R8-S(O)2-. R8 is C 1-4 Selected from alkyl, and q is selected from 0, 1, 2, 3, and 4.

[0044] In any embodiment of the compound of formula IIb of this disclosure, R1 is selected from methyl and R2 is selected from H, Cl, and F.

[0045] In any embodiment of the compound of formula IIb in this disclosure, R1 and R2 are -O-(CH2) m The system is cyclized to -O-, where m is selected from 1 or 2.

[0046] In any embodiment of the compound of formula IIb of this disclosure, R3 and R4 are both H.

[0047] In any embodiment of the compound of formula IIb of this disclosure, R3 is H and R4 is methyl.

[0048] In any embodiment of the compound of formula IIb of this disclosure, X2 is selected from H, OH, NH2, or N(i-Pr)-S(O)2-CH3.

[0049] In any embodiment of the compounds of formula IIb of this disclosure, X2 is selected from H and q is 0, or X2 is selected from OH or -N(R6)(R7) and q is 0, 1, 2, 3 or 4.

[0050] In any embodiment of the compound of formula IIb of this disclosure, X2 is selected from OH or -N(R6)(R7), q is 1, 2, 3 or 4, R3 and R4 are both H, R1 is selected from methyl and R2 is selected from H, Cl or F, or R1 and R2 are -O-(CH2) m The system is cyclized to -O-, where m is selected from 1 or 2.

[0051] In some embodiments of the compounds of formula IIb of this disclosure, the compound of formula IIb is selected from the compounds of formula IIb-1 or formula IIb-2 described below.

[0052] [ka]

[0053] In the compounds of formula IIb-1 and formula IIb-2, R1, R2, X2, and q are as defined in any one embodiment of the compound of formula IIb.

[0054] In some embodiments of the compound of formula IIb, formula IIb is selected from the following compounds or their pharmaceutically acceptable salts or stereoisomers.

[0055] [ka]

[0056] In one embodiment of the present disclosure, the compounds of formula II, formula IIa, formula IIb, formula IIa-1, formula IIb-1, formula IIa-2 and formula IIb-2 do not include the following compounds.

[0057] [ka]

[0058] In another respect, the present disclosure provides pharmaceutical compositions comprising compounds of formula II, formula IIa, formula IIb, formula IIa-1, formula IIb-1, formula IIa-2, or formula IIb-2, or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable excipients.

[0059] In a further respect, the Disclosure provides the use of compounds of formula II, formula IIa, formula IIb, formula IIa-1, formula IIb-1, formula IIa-2 or formula IIb-2, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition thereof, in the manufacture of a pharmaceutical for the treatment of tumors.

[0060] In another aspect, the present disclosure provides a method for treating a tumor, comprising administering to a patient in need an effective amount of the compound of formula II, formula IIa, formula IIb, formula IIa-1, formula IIb-1, formula IIa-2 or formula IIb-2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof.

[0061] In another aspect, the present disclosure provides compounds of formula II, formula IIa, formula IIb, formula IIa-1, formula IIb-1, formula IIa-2 or formula IIb-2, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions used for the treatment of tumors.

[0062] In another aspect, the present disclosure provides the use of compounds of formula II, formula IIa, formula IIb, formula IIa-1, formula IIb-1, formula IIa-2 or formula IIb-2, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions thereof, in the treatment of tumors.

[0063] As is well known to those skilled in the art, antibody-drug conjugates (ADCs) generally cause toxic effects due to the detachment of small amounts of payload in the body. In contrast, the compounds of this disclosure, particularly compound 2, exhibit better stability in plasma, a shorter half-life and lower in vivo exposure, and are more readily eliminated when detached as the ADC payload in the body, resulting in lower toxicity. Furthermore, toxicity studies of the compounds of this disclosure show lower in vivo toxicity at the same dose and demonstrate superior safety whether used alone as a cancer treatment or as a pharmaceutical product for manufacturing ADCs.

[0064] Terminology and Explanation Unless otherwise specified, terms used herein have their general meanings in the art. Unless specifically defined, any particular term or phrase should be understood as having its ordinary definition, not as uncertain or ambiguous. Where a trade name appears herein, it refers to the corresponding product or its active ingredient.

[0065] In any embodiment of the compound of formula II in this disclosure, R1 and R2 are -O-(CH2) m When cyclized to -O-, the compound of formula II is the compound of formula II-1 below. In the compound of formula II of this disclosure or any embodiment thereof, R3 and R4 are -(CH2) k When cyclized, it is represented in the compound form of formula II-2 below.

[0066] [ka]

[0067] The terms "optional" or "optionally" mean that the matters or circumstances described below may or may not occur, and this description includes both cases where the matters or circumstances occur and where they do not. For example, the term "optionally substituted" means that the corresponding group may or may not be substituted, or it may be substituted with a specified substituent, and all cases, with or without substitution, are included in the scope of "optionally".

[0068] The terms "independently selected" or "each independently selected" mean that each of the substituents can select a different option within the specified selection range and is not affected by the selection results of other substituents. If any of the substituents that can be "independently selected" exist in multiple identical substituents in the chemical formula, they can still select the same or different options.

[0069] The term "halogen" or "halo atom" refers to fluorine, chlorine, bromine, and iodine.

[0070] The term "halo" refers to a group formed by replacing one or more hydrogen atoms in a substituent with halogen atoms.

[0071] The term "alkyl" refers to a linear or branched hydrocarbon group in which carbon atoms are linked by single bonds. Alkyl groups are preferably C 1-4 or C 1-6 It is an alkyl group. 1-4 "Alkyl" refers to a linear or branched alkyl group having 1, 2, 3, or 4 carbon atoms. 1-4 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, and tert-butyl. 1-6 "Alkyl" refers to a linear or branched alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms.1-6 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, and n-hexyl.

[0072] The term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are substituted by halogen atoms. 1-4 Examples of haloalkyls include, but are not limited to, trifluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, and 2,2,2-trichloroethyl.

[0073] The term "alkoxy" refers to alkyl-O-, where alkyl is as defined above, C 1-4 Alkoxy or C 1-6 Contains alkoxy. 1-4 Alkoxy is "C 1-4 It can be understood as "alkyl-O-", and examples include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, and tert-butoxy. 1-6 Alkoxy is "C 1-6 It can be understood as "alkyl-O-", and an example of this is the above C 1-4 In addition to specific examples of alkoxys, this includes, but is not limited to, n-pentyloxy, neopentyloxy, and n-hexyloxy.

[0074] The term "haloalkoxy" refers to an alkoxy in which one or more hydrogen atoms are substituted by halogen atoms. 1-4 Examples of haloalkoxys include, but are not limited to, trifluoromethoxy, trichloromethoxy, 2,2,2-trifluoroethoxy, and 2,2,2-trichloroethoxy.

[0075] The term "cycloalkyl" refers to a saturated monocyclic or polycyclic hydrocarbon group in which carbon atoms are linked cyclically by single bonds. The hydrogen atoms of the ring carbon atoms may be optionally substituted with oxo atoms, that is, the "-CH2-" in the ring can be optionally oxo-substituted to form "-C(O)-". 3- to 6-membered cycloalkyl refers to 3, 4, 5, or 6-membered ring cycloalkyls, and specific examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0076] The term "heterocycloalkyl" refers to a cycloalkyl group in which one or more (e.g., two, three, or four) cyclocarbon atoms are substituted by a heteroatom or heteroatomic group (i.e., an atomic group containing a heteroatom). The ring carbon atoms refer to carbon atoms constituting a cyclic skeleton structure, and the heteroatom means an atom other than C and H in an organic compound, such as a nitrogen atom (N), an oxygen atom (O), an sulfur atom (S), an oxygen atom (P), or an or boron atom (B). The heteroatoms, such as nitrogen and sulfur atoms, are oxidized, and the nitrogen atom is quaternized. Examples of the heteroatomic group include, but are not limited to, -S(=O)2-, -S(=O)- and optionally substituted -NH-, -S(=O)(=NH)-, -C(=O)NH-, -C(=NH)-, -S(=O)2NH-, S(=O)NH-, or -NHC(=O)NH-. 3- to 6-membered heterocycloalkyls refer to 3, 4, 5, or 6-membered heterocycloalkyls, and specific examples include, but are not limited to, azilidinyl, oxyranil, thiranil, azetidinil, oxetanil, thietanil, pyrrolidinyl, tetrahydroflapiperidinyl, tetrahydropyranil, morpholinil, and 1,4-dioxanil.

[0077] The term “composition” means including products containing specified amounts of each designated component, and any products resulting directly or indirectly from combinations of each designated component in specified amounts. Those skilled in the art can modify the substantial dose levels of each active component in the pharmaceutical compositions of this disclosure to obtain amounts of the active compound that are effective for specific patients, compositions, and methods of administration and that produce a desired therapeutic response.

[0078] The term "pharmaceutically acceptable" applies to those compounds, materials, compositions and / or dosage forms that are within the bounds of reliable medical judgment, suitable for contact with human and animal tissues, with little toxicity, irritation, allergic reaction or other problem or complication, and that meet a reasonable benefit / risk ratio.

[0079] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt of an acid or base, and includes salts formed by a compound with an inorganic or organic acid, as well as salts formed by a compound with an inorganic or organic base.

[0080] The term "excipient" generally refers to a carrier, diluent, and / or medium necessary for the formulation of an effective pharmaceutical composition.

[0081] The term “effective dose” means an amount of the compound of the Disclosure or a pharmaceutically acceptable salt thereof that is appropriate for a reasonable efficacy-to-risk ratio of the medical treatment to treat a disease or condition. The amount constituting the “effective dose” of the compound of the Disclosure varies depending on the compound, the disease condition and its severity, the method of administration, and the age of the mammal being treated, but may be routinely determined by those skilled in the art based on their knowledge and the Disclosure.

[0082] In this disclosure, in the substituent

[0083] [ka]

[0084] The symbol indicates the position where the substituent connects to the parent structure or other fragments. The dash "-" in the substituent structure is used to indicate the bonding point of the substituent; for example, -CH3 indicates that this group connects to the parent structure or other fragments via a carbon atom.

[0085] [ka]

[0086] and

[0087] [ka]

[0088] This represents the absolute configuration of the center of the solid, i.e., the R or S configuration.

[0089] In this disclosure, chemical bonds are depicted with solid and dotted lines.

[0090] [ka]

[0091] This indicates a single bond or a double bond, which can be determined by a person skilled in the art based on the valence of the atoms at both ends of the chemical bond.

[0092] In this disclosure, “isomer” includes geometric isomers and stereoisomers, such as atropisomers, cis-trans isomers, enantiomers, diastereomers, tautomers, and racemic and other mixtures thereof, all of which are within the scope of this disclosure. The term “enantiomer” refers to a stereoisomer that is a mirror image of one another. The term “tautomer” refers to a type of functional isomer in which the linkage of hydrogens differs due to the movement of one or more double bonds, for example, ketones and their enol forms are keto-enol tautomers. The term “diastereomer” refers to a stereoisomer in which a molecule has two or more chiral centers and the molecules are not mirror images of each other. The term “cis-trans isomer” refers to a different spatial arrangement in which a double bond or single bond of a ring-forming carbon atom in a molecule cannot rotate freely. The term “atropisomer” refers to a stereoisomer that can be separated because the rotation between single bonds is hindered or very slow.

[0093] Stereoisomers of the compounds of this disclosure can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. For example, enantiomers of certain compounds of this disclosure can be prepared by asymmetric catalytic techniques or chiral auxiliary derivatization techniques. Alternatively, compounds having a single stereochemistry can be obtained from a mixture by chiral resolution techniques. Or, they can be obtained by preparation directly from chiral starting materials. Separation of optically pure compounds in this disclosure is usually achieved by preparative chromatography using a chiral column to achieve the objective of separating chiral compounds.

[0094] In this disclosure, the unit M for solution concentration represents mol / L, mM represents mmol / L, and nM represents nmol / L. The unit N for solution concentration represents equivalent concentration, that is, equivalent concentration is expressed as the gram equivalents of solute contained in 1 liter of solution, and is represented by the symbol N. For example, if 1 liter of concentrated hydrochloric acid contains 12.0 gram equivalents of hydrochloric acid (HCl), the concentration is 12.0 N. Equivalent concentration = number of gram equivalents of solute / volume of solution (liters).

[0095] The chemical abbreviations used in this disclosure and the chemical names they refer to are as follows:

[0096] [Table 1]

[0097] [Examples] The following are specific manufacturing examples and biological experiments to further illustrate the present disclosure, but these are for illustrative purposes only and should not be construed as limiting the present disclosure in any way. Those skilled in the art will understand that, unless otherwise specified below, the materials used are well known in the art, available on the market, or obtainable by those skilled in the art based on known literature or by common methods. Unless otherwise stated, all reactions in the present disclosure are carried out in a dry solvent under a dry nitrogen or argon gas atmosphere and continuous magnetic stirring. Herein, (i) the temperature is expressed in degrees Celsius (°C), the operation is carried out at room temperature (generally 15-35°C, preferably 20-30°C, more preferably 20-25°C), (ii) the solvent is removed by vacuum evaporation using a rotary evaporator, the bath temperature not exceeding 60°C, (iii) the progress of the reaction is tracked by thin-layer chromatography (TLC), and (iv) the final product is obtained by obtaining a satisfactory hydrogen nuclear magnetic resonance spectrum ( 1 (H-NMR) and / or mass spectrometry (MS) data are available.

[0098] Example 1: Preparation of Compound 1 Step 1:

[0099] [ka]

[0100] 86.4 mL of 2,2,6,6-tetramethylpiperidine was added to a 3 L three-necked flask, and 1 L of anhydrous tetrahydrofuran was added. The reaction system was cooled to -78°C and reacted for 15 minutes. 272 ​​mL of 2.5 M n-butyllithium solution in petroleum ether was added and reacted for 30 minutes. 32 g of compound L-1 was dissolved in 200 mL of anhydrous tetrahydrofuran, added to the reaction system, and reacted for 1 hour. 50 mL of 1-penten-3-one was dissolved in 200 mL of anhydrous tetrahydrofuran, added to the reaction system, and reacted for 1 hour. After detecting completion of the reaction by TLC, 640 mL of 4N hydrochloric acid was added and extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium bicarbonate aqueous solution, washed with saturated brine, and dried over anhydrous sodium sulfate. After vacuum concentration of the reaction system, 21 g of compound L-2 was obtained by column chromatography with a yield of 49%. ESI-MS m / z: 253.68, [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 6.99 (s, 1H), 6.13 (dd, J = 17.3, 10.6 Hz, 1H), 5.29 (d, J = 7.0 Hz, 1H), 5.26 (s, 1H), 3.99 (s, 3H), 2.73 (s, 2H), 2.04 (dt, J = 14.8, 7.3 Hz, 1H), 1.98 - 1.87 (m, 1H), 0.91 (t, J = 7.3 Hz, 3H).

[0101] Step 2

[0102] [ka]

[0103] 21 g of compound L-2 was added to a 1 L three-necked flask, and 400 mL of anhydrous ethanol was added. After cooling the reaction system to 0°C, 14.21 g of sodium borodeuteride was added, and the mixture was stirred for 15 minutes. The temperature was then raised to room temperature and the reaction was allowed to proceed for 12 hours. After monitoring the completion of the reaction by TLC, 30 mL of 1 N hydrochloric acid was added, and the mixture was extracted with dichloromethane. The organic phases were combined and sequentially washed with saturated sodium bicarbonate aqueous solution, saturated brine, and dried over anhydrous sodium sulfate. After vacuum concentration of the reaction system, 17 g of compound L-3 was obtained as a clear viscous liquid by column chromatography with a yield of 80%. ESI-MS m / z: 259.73, [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 6.99 (s, 1H), 6.13 (dd, J = 17.3, 10.6 Hz, 1H), 5.29 (d, J = 7.0 Hz, 1H), 5.26 (s, 1H), 3.99 (s, 3H), 2.73 (s, 2H), 2.04 (dt, J = 14.8, 7.3 Hz, 1H), 1.98 - 1.87 (m, 1H), 0.91 (t, J = 7.3 Hz, 3H).

[0104] Step 3:

[0105] [ka]

[0106] 2 g of compound L-3 was added to a 250 mL round-bottom flask, and 100 mL of dichloromethane was added. The reaction system was cooled to -78 °C, ozone was introduced, and after detecting completion of the reaction by TLC, 0.5 mL of dimethyl sulfide was added, and the mixture was heated to room temperature and stirred for 30 minutes. After vacuum concentration of the reaction system, 1.9 g of compound L-4 was obtained by column chromatography with a yield of 95%. The product was used directly in the next reaction without further separation or purification. ESI-MS m / z: 261.70, [M+H] + . 1H NMR (400 MHz, CDCl3) δ 7.15 (s, 1H), 5.19 (d, J = 4.8 Hz, 1H), 3.96 (s, 3H), 3.21 (d, J = 4.8 Hz, 1H), 2.64 (s, 1H), 1.80 (q, J = 7.5 Hz, 2H), 0.92 (t, J = 7.5 Hz, 3H).

[0107] Step 4:

[0108] [ka]

[0109] 12 g of compound L-4 was added to a 1 L necked flask, 240 mL of dichloromethane was added, and the mixture was stirred for 30 minutes. The reaction system was then cooled to 0°C. 286 mg of 2,2,6,6-tetramethylpiperidinium oxide, 616 mg of sodium bicarbonate, 654 mg of potassium bromide, and 18 mL of water were added, and the mixture was stirred for 15 minutes. 120 mL of aqueous sodium hypochlorite solution (>7.5 wt%) was added, and the mixture was reacted for 30 minutes. After monitoring the completion of the reaction by TLC, 12 g of sodium bisulfite was added, and the mixture was extracted with dichloromethane. The organic phases were combined, washed with water, washed with saturated brine, and dried over anhydrous sodium sulfate. After vacuum concentration of the reaction system, 6.6 g of compound L-5 was obtained by column chromatography, with a yield of 55%. ESI-MS m / z: 259.68, [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 7.20 (s, 1H), 4.00 (s, 3H), 3.64 (s, 1H), 1.79 (q, J = 7.4 Hz, 2H), 0.97 (t, J = 7.4 Hz, 3H).

[0110] Step 5:

[0111] [ka]

[0112] Under a carbon monoxide atmosphere, 3 g of compound L-5 was added to a 100 mL three-necked flask, and 286 mg of 1,3-bis(diphenylphosphin)propane, 2.4 g of potassium carbonate, 129 mg of palladium acetate, 15 mL of N,N-dimethylformamide, and 30 mL of deuterated methanol were added. The reaction system was heated to 65 °C and reacted for 12 hours. After monitoring the completion of the reaction by TLC, 20 mL of 1 N hydrochloric acid was added, and the mixture was extracted with ethyl acetate. The organic phases were combined and sequentially washed with saturated sodium bicarbonate aqueous solution and saturated brine, and dried over anhydrous sodium sulfate. After vacuum concentration of the reaction system, 1.33 g of compound L-6 was obtained by column chromatography, yielding 40%. ESI-MS m / z: 286.29, [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 7.98 (d, J = 2.0 Hz, 1H), 4.09 (d, J = 2.0 Hz, 3H), 3.71 (d, J = 2.6 Hz, 1H), 1.88 - 1.75 (m, 2H), 0.96 (td, J = 7.3, 1.9Hz, 3H).

[0113] Step 6:

[0114] [ka]

[0115] 1 g of compound L-6 was added to a 50 mL two-necked flask, 1.05 g of sodium iodide and 10 mL of acetonitrile were added, and the reaction system was cooled to 0°C and stirred for 30 minutes. 0.89 mL of trimethylsilanclide was added, and the reaction system was raised to room temperature and reacted for 12 hours. After monitoring the completion of the reaction by TLC, 40 mL of water and 1 mL of saturated sodium bisulfite aqueous solution were added, and the mixture was stirred for 1 hour. The mixture was extracted with dichloromethane and dried over anhydrous sodium sulfate. After vacuum concentration of the reaction system, it was purified by column chromatography to obtain 800 mg of compound L-7, with a yield of 84%. ESI-MS m / z: 272.27, [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 10.03 (s, 1H), 7.32 (s, 1H), 3.90 (s, 1H), 1.81 (dd, J = 14.5, 7.2 Hz, 2H), 0.99 (t, J = 7.4 Hz, 3H).

[0116] Step 7:

[0117] [ka]

[0118] 620 mg of compound L-7 was added to a 25 mL two-necked flask, and 1.48 g of cesium carbonate, 8 mL of dimethyl sulfoxide, and 1.67 mL of tert-butyl acrylate were added. The reaction system was heated to 50 °C and reacted for 12 hours. After monitoring the completion of the reaction by TLC, 1 mL of concentrated hydrochloric acid and 40 mL of water were added, and the mixture was extracted with dichloromethane and dried over anhydrous sodium sulfate. After vacuum concentration of the reaction system, 468 mg of compound L-8 was obtained by preparative SPC chiral separation (instrument model: SFC-350 (Waters), chromatography column: AS 25 × 250 mm, 10 μm, mobile phase: CO2 / MEOH = 65 / 35, flow rate: 200 ml / min), yielding 55%. ESI-MS m / z: 365.38, [M+H] + .

[0119] Step 8:

[0120] [ka]

[0121] 200 mg of compound L-8 was added to a 10 mL necked flask, 5 mL of toluene and 0.5 mL of trifluoroacetic acid were added, and the reaction system was heated to 110°C and reacted for 2 hours. After monitoring the completion of the reaction by TLC, the reaction system was concentrated under vacuum and purified by column chromatography to obtain 140 mg of compound L-9, with a yield of 98%. ESI-MS m / z: 265.26, [M+H] + .

[0122] Step 9:

[0123] [ka]

[0124] 300 mg of compound L-9 was added to a 25 mL necked flask, and 266 mg of N-(5-methyl-6-fluoro-8-amino-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)acetamide, 9 mL of toluene, 770 mL of o-cresol, and 133 mg of pyridinium p-toluenesulfonate were added. The reaction system was heated to 110°C and reacted for 18 hours. After monitoring the completion of the reaction by TLC, the reaction system was concentrated under vacuum and purified by column chromatography (dichloromethane:methanol = 10:1) to obtain 443 mg of compound L-10, with a yield of 81%.

[0125] Step 10:

[0126] [ka]

[0127] 443 mg of intermediate compound L-10 was added to a 25 mL necked flask, followed by 7 mL of purified water and 2.2 mL of methanesulfonic acid. The reaction system was then purged with a nitrogen atmosphere. The reaction system was heated to 85°C and allowed to react for 10 hours. After monitoring the completion of the reaction by TLC, the reaction system was concentrated under vacuum and purified by column chromatography (dichloromethane:methanol) to obtain 141 mg of compound 1, with a yield of 35%. ESI-MS m / z: 437.47, [M+H] + . 1 H NMR (400 MHz, D2O) δ 7.20 - 7.06 (m, 2H), 5.42 - 5.14 (m, 5H), 3.30 (dd, J = 18.1, 4.0 Hz, 1H), 3.06 - 2.91 (m, 1H), 2.69 (s, 2H), 2.62 - 2.47 (m, 1H), 2.18 (s, 3H), 1.80 (q, J = 7.3 Hz, 2H), 0.79 (t,J = 7.4 Hz, 3H).

[0128] Example 2: Preparation of Compound 2

[0129] [ka]

[0130] 2.4 mL of water and 3.0 mL of tetrahydrofuran were added to a mixture of anhydrous sodium sulfate (2.25 eq), 2-oxymesyanoethyl acetate (2.25 eq), 2-hydroxyacetic acid (1.45 eq), and methanesulfonate of compound 1 (100 mg). The mixture was stirred at room temperature for 15 minutes, then N-methylmorpholine (1.10 eq) was added and the mixture was stirred at room temperature for 15 minutes. Finally, EDCI (2 eq) was added and the mixture was stirred at room temperature for 3 hours. The completion of the reaction of compound 1 was monitored by HPLC, the mixture was filtered, the cake was dried, and the compound was separated and purified by silica gel column chromatography (DCM:MeOH) to obtain the title compound.

[0131] 1H NMR (400 MHz, DMSO) δ 8.41 (d, J = 8.9 Hz, 1H), 7.77 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.58 (d, J = 5.4 Hz, 1H), 5.49 (t, J = 5.7 Hz, 1H), 5.20 (t, J = 13.9 Hz, 2H), 3.96 (d, J = 5.5 Hz, 2H), 3.28 - 3.05 (m, 2H), 2.39 (s, 3H), 2.29 - 2.11 (m, 2H), 1.96 - 1.78 (m, 2H), 0.87 (t, J = 7.2 Hz, 3H). MS (ESI) m / z [M+H] + : 496.1.

[0132] Example 3: Preparation of Compound 3

[0133] [ka]

[0134] Referring to the manufacturing method of Example 2, (R)-2-cyclopropyl-2-hydroxyacetic acid was used as a starting material to obtain 52 mg of compound 3, with a yield of 52%.

[0135] 1H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J = 8.4 Hz, 1H), 7.78 (d, J = 10.9 Hz, 1H), 7.31 (s, 1H), 6.52 (s, 0H), 5.58 - 5.48 (m, 1H), 5.38 (d, J = 5.3 Hz, 1H), 5.21 (q, J = 19.0 Hz, 2H), 3.62 (d, J = 5.2 Hz, 1H), 3.16 (dd, J = 11.2, 5.7 Hz, 3H), 2.39 (s, 4H), 2.22 - 2.12 (m, 3H), 1.86 (dt, J = 14.3, 7.1 MS (ESI) m / z [M+H] + : 536.2.

[0136] Example 4: Preparation of Compound 4

[0137] [ka]

[0138] Referring to the manufacturing method of Example 2, (S)-2-cyclopropyl-2-hydroxyacetic acid was used as a starting material to obtain 54 mg of compound 4, with a yield of 54%.

[0139] 1H NMR (400 MHz, DMSO-d6) δ 8.37 (d, J = 9.0 Hz, 1H), 7.77 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.51 (s, 1H), 5.58 (q, J = 6.7 Hz, 1H), 5.49 (d, J = 5.2 Hz, 1H), 5.35 - 5.06 (m, 2H), 3.60 (t, J = 5.7 Hz, 1H), 3.17 (q, J = 17.2 Hz, 2H), 2.39 (s, 3H), 2.16 (d, J = 6.1 Hz, 2H), 1.85 (dq, J = 14.2, 7.1 Hz, MS (ESI) m / z [M+H] + : 536.2.

[0140] Example 5: Preparation of Compound 5

[0141] [ka]

[0142] Referring to the manufacturing method of Example 2, (R)-3-hydroxybutyric acid was used as a raw material to obtain 55 mg of compound 5, with a yield of 56%.

[0143] 1H NMR (400 MHz, DMSO-d6) δ 8.42 (d, J = 8.7 Hz, 1H), 7.78 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.55 (dt, J = 9.3, 5.0 Hz, 1H), 5.29 - 5.12 (m, 2H), 4.66 (d, J = 4.7 Hz, 1H), 4.05 (dt, J = 12.1, 6.0 Hz, 1H), 3.25 - 3.09 (m, 2H), 2.39 (s, 3H), 2.33 - 2.17 (m, 2H), 2.13 (dd, J = 10.7, MS (ESI) m / z [M+H] + : 524.2.

[0144] Example 6: Preparation of Compound 6

[0145] [ka]

[0146] Referring to the manufacturing method of Example 2, (S)-3-hydroxybutyric acid was used as a raw material to obtain 51 mg of compound 6, with a yield of 52%.

[0147] 1H NMR (400 MHz, DMSO-d6) δ 8.41 (d, J = 8.7 Hz, 1H), 7.78 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.57 (dt, J = 8.8, 4.5 Hz, 1H), 5.22 (d, J = 2.7 Hz, 2H), 4.64 (d, J = 4.5 Hz, 1H), 4.05 (dt, J = 11.9, 6.1 Hz, 1H), 3.16 (t, J = 5.7 Hz, 2H), 2.39 (s, 3H), 2.32 - 2.17 (m, 2H), 2.17 - 2.04 (m, 2H), 1.86 (hept, J = 7.1 Hz, 2H), 1.08 (d, J = 6.1 Hz, 3H), 0.87 (t, J = 7.3 Hz, 3H). MS (ESI) m / z [M+H] + : 524.2.

[0148] Example 7: Preparation of Compound 7

[0149] [ka]

[0150] Compound 7-1 (87 mg, 1 eq), Compound 7-2 (144 mg, 1.1 eq), pyridinium p-toluenesulfonate (50 mg, 0.4 eq), acetic acid (2.6 mL, 30V), and toluene (2.6 mL, 30V) were placed in a reaction flask and reacted at 110°C for 18 hours. After monitoring the completion of the reaction by TLC, the solvent was rotated and evaporated, and the mixture was separated and purified by silica gel column chromatography (dichloromethane:methanol) to obtain 123 mg of Compound 7, with a yield of 61%.

[0151] 11H NMR (400 MHz, DMSO-d6) δ 7.72 (d, J = 9.0 Hz, 1H), 7.29 (d, J = 9.0 Hz, 1H), 7.17 (s, 1H), 6.45 (s, 1H), 5.65 (s, 2H), 5.16 (s, 2H), 3.04 (t, J = 6.1 Hz, 2H), 2.75 (t, J = 6.1 Hz, 2H), 2.08 - 1.95 (m, 2H), 1.85 (hept, J = 7.1 Hz, 2H), 0.88 (t, J = 7.2 Hz, 3H). MS (ESI) m / z [M+H] + : 406.1。

[0152] Example 8 Preparation of Compound 8

[0153]

Chem.

[0154] Referring to the production method of Example 7, using Compound 8-1 and 7-2 as raw materials, 127 mg of Compound 8 was obtained, and the yield was 60%.

[0155] 1H NMR (400 MHz, DMSO-d6) δ 8.41 (d, J = 8.7 Hz, 1H), 7.78 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.57 (dt, J = 8.8, 4.5 Hz, 1H), 5.22 (d, J = 2.7 Hz, 2H), 4.64 (d, J = 4.5 Hz, 1H), 4.05 (dt, J = 11.9, 6.1 Hz, 1H), 3.16 (t, J = 5.7 Hz, 2H), 2.39 (s, 3H), 2.32 - 2.17 (m, 2H), 2.17 - 2.04 (m, 2H), 1.86 (hept, J = 7.1 Hz, 2H), 1.08 (d, J = 6.1 Hz, 3H), 0.87 (t, J = 7.3 Hz, 3H). MS (ESI) m / z [M+H] + : 424.1.

[0156] Example 9: Preparation of Compound 9 Step 1:

[0157] [ka]

[0158] 250 mg of compound 9-1, 300 mg of compound 7-2, 7.5 mL of acetic acid, 7.5 mL of toluene, and 68 mg of pyridinium p-toluenesulfonate were added to a 50 mL necked flask and reacted at 110°C for 18 hours. After monitoring the completion of the reaction by TLC, the solvent was evaporated under reduced pressure, and the compounds were separated and purified by silica gel column chromatography (DCM:MeOH) to obtain 386 mg of compound 9-3, with a yield of 81%. MS (ESI) m / z [M+H] + :480.2.

[0159] Step 2:

[0160] [ka]

[0161] 380 mg of compound 9-3 was added to a 100 mL necked flask, 7.6 mL of purified water and 3.8 mL of methanesulfonic acid were added, and the mixture was reacted at 85°C for 10 hours under the protection of nitrogen gas. After monitoring the completion of the reaction by TLC, the mixture was cooled to room temperature, 22.8 mL of methanol was added, and the mixture was stirred at room temperature for 2 hours. The crude product was filtered, separated and purified by silica gel column chromatography (dichloromethane:methanol) to obtain 148 mg of compound 9, with a yield of 35%.

[0162] 1H NMR (400 MHz, D2O) δ 7.20-7.06 (m, 2H), 5.42-5.14 (m, 5H), 3.30 (dd, J = 18.1, 4.0 Hz, 1H), 3.06-2.91 (m, 1H), 2.69 (s, 2H), 2.62-2.47 (m, 1H), 2.18 (s, 3H), 1.80 (q, J = 7.3 Hz, 2H), 0.79 (t, J = 7.4 Hz, 3H). MS (ESI) m / z [M+H]+: 438.2.

[0163] Example 10: Preparation of Compound 10

[0164] [ka]

[0165] Referring to the manufacturing method in Example 9, compound 10-1 and compound 7-2 were used as raw materials to obtain 70 mg of compound 10, with a yield of 34%.

[0166] 1H NMR (400 MHz, DMSO-d6) δ 8.32 (d, J = 8.4 Hz, 1H), 8.20 (dd, J = 8.4, 1.2 Hz, 1H), 7.93-7.84 (m, 1H), 7.79 (t, J = 7.6 Hz, 1H), 7.35 (s, 1H), 6.56 (s, 1H), 5.42 (s, 2H), 3.98 (p, J = 6.7 Hz, 1H), 3.50 (t, J = 8.0 Hz, 2H), 3.42-3.35 (m, 2H), 3.00 (s, 3H), 1.88 (hept, J = 7.3 Hz, 2H), 1.15 (d, J = 6.7 Hz, 6H), 0.88 (t, J = 7.3 Hz, 3H). MS (ESI) m / z [M+H] + : 514.2.

[0167] Example 11 Production of Compound 11

[0168]

change

[0169] According to the manufacturing method of Example 9, the raw materials of Compound 11-1 and Compound 7-2 were used, and 80 mg of Compound 11 was obtained with a yield of 37%.

[0170] 1 H NMR (500 MHz, DMSO-d6) δ 0.88 (t, J = 7.25 Hz, 3H), 1.32 (t, J = 7.5 Hz, 3H), 1.85 (m, 2H), 3.11 (q, J = 7.5 Hz, 2H), 5.26(s, 2H), 6.48 (s, 1H), 7.23 (s, 1H), 7.41 (d, J = 10.0 Hz, 2H), 8.01 (d, J = 10.0 Hz, 1H), 10.3 (s, 1H). MS (ESI) m / z [M+H] + : 395.1.

[0171] Example 12 Production of Compound 12

[0172] [ka]

[0173] 255 mg of compound 12-1, 300 mg of compound 7-2, 7.5 mL of acetic acid, 7.5 mL of toluene, and 68 mg of pyridinium p-toluenesulfonate were added to a 50 mL necked flask and reacted at 110°C for 18 hours. After monitoring the completion of the reaction by TLC, the solvent was evaporated under reduced pressure, and the compounds were separated and purified by silica gel column chromatography (DCM:MeOH) to obtain 330 mg of compound 12-3, with a yield of 70%.

[0174] 1 H NMR (400 MHz, DMSO-d6) δ 7.63 (s, 1H), 7.51(s, 1H), 7.24 (s, 1H), 6.50 (s, 1H), 6.30 (s, 2H), 5.26 (s, 2H), 3.81 (d, J = 5.9 Hz, MS (ESI) m / z [M+H] + :471.1.

[0175] [ka]

[0176] 300 mg of intermediate compound 12-3 was dissolved in 15 mL of 10% sulfuric acid and reacted at 110°C for 48 hours. After monitoring the completion of the reaction by TLC, saturated sodium carbonate aqueous solution was added to adjust the pH to neutral, and the mixture was extracted with dichloromethane. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (DCM:MeOH) to obtain 89 mg of compound 12, with a yield of 31%.

[0177] 1H NMR (400 MHz, DMSO-d6) δ 7.63 (s, 1H), 7.50(s, 1H), 7.24 (s, 1H), 6.48 (s, 1H), 6.28 (s, 2H), 5.32-5.19 (m, 2H), 3.51-3.46 (m, MS (ESI) m / z [M+H] + : 453.2.

[0178] Example 13: Preparation of Compound 13

[0179] [ka]

[0180] Referring to the manufacturing method of Example 12, compound 13-1 and compound 7-2 were used as raw materials to obtain 74 mg of compound 13, with a two-step yield of 16%.

[0181] 1 H NMR (400 MHz, DMSO-d6) δ 7.63 (s, 1H), 7.50(s, 1H), 7.24 (s, 1H), 6.48 (s, 1H), 6.28 (s, 2H), 5.32-5.19 (m, 2H), 3.51-3.46 (m, MS (ESI) m / z [M+H] + : 467.2.

[0182] Example 14: Preparation of Compound 14 Step 1:

[0183] [ka]

[0184] 213 mg of compound 14-1, 300 mg of compound 7-2, 7.5 mL of acetic acid, 7.5 mL of toluene, and 68 mg of pyridinium p-toluenesulfonate were added to a 50 mL necked flask and reacted at 110°C for 18 hours. After monitoring the completion of the reaction by TLC, the solvent was evaporated by rotation, and the compounds were separated and purified by silica gel column chromatography (DCM:MeOH) to obtain 300 mg of compound 14-3 in yield: 68%. MS (ESI) m / z [M+H]+: 443.1.

[0185] Step 2:

[0186] [ka]

[0187] 300 mg of compound 14-3 was dissolved in 3 mL of N,N-dimethylformamide, sodium azide (48 mg, 1.1 eq) was added, and the reaction was carried out at 80°C for 16 hours. After monitoring the completion of the reaction by TLC, the mixture was cooled to room temperature, 18 mL of water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with water, and the organic phase was dried over anhydrous sodium sulfate. The mixture was then filtered and spin-dried to obtain 186 mg of crude compound 14-4, with a yield of 63%. MS (ESI) m / z [M+H]+: 436.1.

[0188] Step 3:

[0189] [ka]

[0190] 186 mg of Compound 14-4 was dissolved in 4 mL of tetrahydrofuran, triphenylphosphine (134 mg, 1.2 eq) was added, and the mixture was stirred at room temperature for 4 hours. After monitoring the completion of the reaction by TLC, hydrochloric acid (4 M, 1 mL) was added to the reaction, and the reaction was carried out at 55 °C for 16 hours. The reaction mixture was concentrated under reduced pressure and separated and purified by a reverse-phase column (SEPA FLASH SW025, Spherical C18, 20 - 45 μm, 100 Å, mobile phase A: 0.05% formic acid / water, mobile phase B: acetonitrile; mobile phase A: mobile phase B = 40:60, flow rate: 20 mL / min) to obtain 63 mg of Compound 14 with a yield of 35%.

[0191] 1 H NMR (400 MHz, DMSO-d6) δ 8.62 (s, 2H), 8.45 (s, 1H), 7.50 (s, 1H), 7.25 (s, 1H), 6.48 (s, 1H), 6.28 (s, 2H), 5.41 (s, 2H), 5.20 (s, 2H), 4.70 (d, J = 6.0 Hz, 2H), 1.86 (tt, J = 14.2, 6.1 Hz, 2H), 0.88 (t, J = 7.2 Hz, 3H). MS (ESI) m / z [M+H] + : 424.1。

[0192] Example 15 Preparation of Compound 15

[0193]

Chem.

[0194] Referring to the preparation method of Example 14, using Compound 15-1 and 7-2 as starting materials, 50 mg of Compound 15 was obtained with a three-step yield of 11%.

[0195] 1H NMR (400 MHz, DMSO-d6) δ 8.62 (s, 2H), 8.45 (s, 1H), 7.50 (s, 1H), 7.25 (s, 1H), 6.48 (s, 1H), 6.28 (s, 2H), 5.41 (s, 2H), 5.20 (s, 2H), 3.51-3.46 (m, 2H), 3.17-3.13 (m, 2H), 1.86 (tt, J = 14.2, 6.1 Hz, 2H), 0.88 (t, J = 7.2 Hz, 3H). MS (ESI) m / z [M+H] + : 438.2.

[0196] Example 16: Preparation of Compound 16

[0197] [ka]

[0198] 1 g of compound 16-1, 1.5 g of compound 7-2, 30 mL of acetic acid, 30 mL of toluene, and 340 mg of pyridinium p-toluenesulfonate were added to a 250 mL necked flask and reacted at 110 °C for 18 hours. After monitoring the completion of the reaction by TLC, the solvent was evaporated by rotation, and the mixture was separated and purified by silica gel column chromatography (dichloromethane:methanol) to obtain 1.6 g of target intermediate compound 16-3 in yield: 75%. MS (ESI) m / z [M+H]+: 431.1.

[0199] [ka]

[0200] 1.6 g of compound 16-3 was dissolved in 16 mL of N,N-dimethylformamide, sodium azide (256 mg, 1.1 eq) was added, and the reaction was carried out at 80°C for 16 hours. After monitoring the completion of the reaction by TLC, the mixture was cooled to room temperature, 96 mL of water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with water, and the organic phase was dried over anhydrous sodium sulfate. The mixture was then filtered and spin-dried to obtain 1.1 g of crude compound 16-4, with a yield of 68%. MS (ESI) m / z [M+H]+: 438.2.

[0201] [ka]

[0202] 1.1 g of compound 16-4 was dissolved in 22 mL of tetrahydrofuran, triphenylphosphine (0.79 g, 1.2 eq) was added, and the mixture was stirred at room temperature for 4 hours. After monitoring the completion of the reaction by TLC, hydrochloric acid (4 M, 5 mL) was added to the reaction system, and the mixture was reacted at 55 °C for 16 hours. The mixture was concentrated under reduced pressure and separated and purified by reverse-phase column chromatography (instrument model: SEPA FLASH SW025, chromatography column: Spherical C18, 20-45 μm, 100 Å, mobile phase A: 0.05% formic acid / water, mobile phase B: acetonitrile, mobile phase A: mobile phase B = 45:55, flow rate 20 mL / min) to obtain 0.58 g of compound 16, with a yield of 56%.

[0203] 1 H NMR (400 MHz, DMSO-d6) δ 8.62 (s, 3H), 8.43 (d, J = 8.0 Hz, 1H), 7.99 (d, J = 10.7 Hz, 1H), 7.35 (s, 1H), 5.59 (s, 2H), 4.70 (d, J = 6.0 MS (ESI) m / z [M+H]+: 412.2.

[0204] Example 17: Preparation of Compound 17

[0205]

Chem.

[0206] 2.4 mL of water and 3.0 mL of tetrahydrofuran were added to a mixture of anhydrous sodium sulfate (2.25 eq), 2-hydroxyacetic acid (1.45 eq) and compound 16 (100 mg). After stirring at room temperature for 15 minutes, N-methylmorpholine (1.10 eq) was added. After stirring at room temperature for 15 minutes, EDCI (2 eq) was added and the mixture was stirred at room temperature for 3 hours. After monitoring the completion of the reaction of compound 16 by HPLC, the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, the solvent was rotary evaporated, and separated and purified by silica gel column chromatography (DCM:MeOH) to obtain 82 mg of compound 17, with a yield of 72%.

[0207] 1 H NMR (400 MHz, DMSO) δ 8.41 (d, J = 8.9 Hz, 1H), 7.77 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.58 (d, J = 5.4 Hz, 1H), 5.49 (t, J = 5.7 Hz, 1H), 5.20 (t, J = 13.9 Hz, 2H), 3.96 (d, J = 5.5 Hz, 2H), 2.55 (s, 3H), 1.96 - 1.78 (m, 2H), 0.87 (t, J = 7.2 Hz, 3H). MS (ESI) m / z [M+H] + : 470.2.

[0208] Example 18 Preparation of Compound 18

[0209]

Chem.

[0210] Referring to the manufacturing method in Example 17, compound 3-1 and compound 16 were used as starting materials to obtain 78 mg of compound 18, with a yield of 63%.

[0211] 1 H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J = 8.4 Hz, 1H), 7.78 (d, J = 10.9 Hz, 1H), 7.31 (s, 1H), 6.52 (s, 0H), 5.58 - 5.48 (m, 1H), 5.38 (d, J = 5.3 Hz, 1H), 5.21 (q, J = 19.0 Hz, 2H), 3.62 (d, J = 5.2 Hz, 1H), 2.39 (s, 4H), 1.86 (dt, J = 14.3, 7.1 Hz, 2H), 1.14 (q, J = 7.3 Hz, 2H), 0.87 (t, J = 7.2 Hz, 3H), 0.38 (dd, J = 20.9, 5.6 Hz, 4H). MS (ESI) m / z [M+H] + : 510.2.

[0212] Example 19: Preparation of Compound 19

[0213] [ka]

[0214] Referring to the manufacturing method in Example 17, compound 4-1 and compound 16 were used as starting materials to obtain 74 mg of compound 19, with a yield of 60%.

[0215] 1H NMR (400 MHz, DMSO-d6) δ 8.37 (d, J = 9.0 Hz, 1H), 7.77 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.51 (s, 1H), 5.58 (q, J = 6.7 Hz, 1H), 5.49 (d, J = 5.2 Hz, 1H), 5.35 - 5.06 (m, 2H), 3.60 (t, J = 5.7 Hz, 1H), 2.39 (s, 3H), 1.85 (dq, J = 14.2, 7.1 Hz, 2H), 1.24 (q, J = 8.1 Hz, 1H), 0.87 (t, J = 7.3 Hz, 3H), 0.59 - 0.32 (m, 4H). MS (ESI) m / z [M+H] + : 510.2.

[0216] Example 20: Preparation of Compound 20

[0217] [ka]

[0218] Referring to the manufacturing method in Example 17, compound 5-1 and compound 16 were used as starting materials to obtain 70 mg of compound 20, with a yield of 58%.

[0219] 1H NMR (400 MHz, DMSO-d6) δ 8.42 (d, J = 8.7 Hz, 1H), 7.78 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.55 (dt, J = 9.3, 5.0 Hz, 1H), 5.29 - 5.12 (m, 2H), 4.66 (d, J = 4.7 Hz, 1H), 4.05 (dt, J = 12.1, 6.0 Hz, 1H), 2.39 (s, 3H), 2.13 (dd, J = 10.7, 5.4 Hz, 2H), 1.86 (hept, J = 7.1 Hz, 2H), 1.09 (d, J = 6.2 Hz, 3H), 0.87 (t, J = 7.3 Hz, 3H). MS (ESI) m / z [M+H] + : 498.2.

[0220] Example 21: Preparation of Compound 21

[0221] [ka]

[0222] Referring to the manufacturing method in Example 17, compound 6-1 and compound 16 were used as starting materials to obtain 75 mg of compound 21, with a yield of 62%.

[0223] 1H NMR (400 MHz, DMSO-d6) δ 8.41 (d, J = 8.7 Hz, 1H), 7.78 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.57 (dt, J = 8.8, 4.5 Hz, 1H), 5.22 (d, J = 2.7 Hz, 2H), 4.64 (d, J = 4.5 Hz, 1H), 4.05 (dt, J = 11.9, 6.1 Hz, 1H), 2.39 (s, 3H), 2.17 - 2.04 (m, 2H), 1.86 (hept, J = 7.1 Hz, 2H), 1.08 (d, J = 6.1 Hz, 3H), 0.87 (t, J = 7.3 Hz, 3H). MS (ESI) m / z [M+H] + : 498.2.

[0224] Bioactivity Test In vitro antitumor activity assay of the payload compound disclosed herein Objective of the study: To verify the in vitro inhibitory activity of the compounds disclosed herein against NCI-N87 (human gastric cancer cells), Calu-3 (human lung adenocarcinoma cells), MDA-MB-453 (human breast cancer cells), KPL-4 (human breast cancer cells), or MDA-MB-468 (human breast cancer cells).

[0225] Logarithmically proliferating tumor cells NCI-N87 (derived from ATCC, product number: CRL-5822), Calu-3 (derived from ATCC, product number: HTB-55), MDA-MB-453 (derived from ATCC, product number: HTB-131), KPL-4 (derived from Nanjing Cobioer, product number: CBP60379), or MDA-MB-468 (derived from ATCC, product number: HTB-132) were selected, added to cell plates at a rate of 5000 cells / well, and cultured in a 37°C, 5% CO2 cell culture incubator for 12-16 hours. 100 μl of sample (starting at 10 μM, 4-fold dilution, 10 concentrations) was added to each well, gently shaken and mixed, and then cultured in the incubator. After 48 hours of culture, 70 μL of CellTiter-Glo was added. TM(Promega, product number: G7572) After adding the working solution and gently shaking to lyse the cells, the data was read using a microplate reader. The formula for calculating the cell proliferation inhibition rate is: Cell proliferation inhibition rate = (1 - sample well / control well) × 100%. Using GraphPad Prism 8.0 software, the logarithm value of the sample concentration was plotted on the x-axis and the cytotoxicity% on the y-axis. Nonlinear regression (curve fitting) analysis was performed on the data to determine the IC for each sample. 50 The values ​​were obtained, and the specific results are shown in Tables 1-1 and 1-2.

[0226] [Table 2]

[0227] [Table 3]

[0228] Experimental results showed that the compounds of this disclosure exhibited significant growth inhibitory activity against NCI-N87, Calu-3, MDA-MB-453, KPL-4, or MDA-MB-468.

[0229] 2. Pharmacokinetics of the compounds of this disclosure in mice Experimental objective: To evaluate the metabolic kinetic properties of the compounds disclosed herein in mice.

[0230] Using Balb / c mice, 12 mice per group were randomly divided into half male and half female groups. A single dose of the sample (Preparation A: 5% DMSO + 95% (15% sulfobutyl-β-cyclodextrin-physiological saline), Preparation B: 5% DMSO-10% Solutol HS 15-85% physiological saline, diluted according to the dose) was administered. Cross-collection points were established, and plasma was collected before administration and at 5 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, and 24 h after administration. The sample concentration in the plasma was measured by LC-MS, and pharmacokinetic parameters were calculated, as shown in Table 2-1.

[0231] [Table 4]

[0232] The experimental results show that the compounds of this disclosure have a short in vivo semi-decay period in mice, and when these compounds are used as payload compounds for ADCs, the ADCs are rapidly removed when payload detachment occurs in the body, resulting in superior in vivo safety.

[0233] 3. Pharmacokinetics of the compounds disclosed herein in rats Experimental objective: To evaluate the metabolic kinetic properties of the compounds disclosed herein in rats.

[0234] SD rats (Beijing Vital River Laboratory Animal Technology Co., Ltd.) were used, and the rats were randomly divided into groups of 6 rats each, with half male and half female. A single dose of the sample (Preparation A: 5% DMSO + 95% (15% sulfobutyl-β-cyclodextrin-physiological saline), diluted according to the dose) was administered, cross-collection points were established, and plasma was collected before administration and at 5 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, and 24 h after administration. The sample concentration in the plasma was measured by LC-MS, and pharmacokinetic parameters were calculated. The results are shown in Table 3-1.

[0235] [Table 5]

[0236] Experimental results showed that the compounds of this disclosure exhibited a relatively short in vivo ptosis period and rapid in vivo clearance in rats.

[0237] 4. Extracorporeal plasma stability of the compounds disclosed herein. 398 μL of plasma was collected, preheated to 37°C, and incubated together for 15 minutes. 2 μL of the sample solution was added to the plasma to adjust the final concentration of the sample to 5 μM. The above solutions were incubated together at 37°C, and 50 μL of the solution was collected at 0, 15, 30, 60, and 120 minutes, and added to 450 μL of ice-cold acetonitrile (containing an internal standard). After vortexing the solutions for 10 minutes, they were centrifuged, and the residual content of the original substrate was detected by LC-MS / MS. See Table 4-1 for specific results.

[0238] [Table 6]

[0239] Experimental results indicate that the compounds of this disclosure have good plasma stability.

[0240] 5. Repeated-dose toxicity studies in rats of the compounds disclosed herein. Preparation of test samples: The test samples were weighed into glass bottles according to the specified weight. First, 1 mL of DMSO was added and mixed uniformly. After the test samples had dissolved, 1 mL of Solutol HS-15 was added and mixed. Then, 8 mL of sodium chloride injection solution was added and mixed by vortexing to obtain a clear and transparent solution.

[0241] SD rats (Beijing Vital River Laboratory Animal Technology Co., Ltd.) were used as test animals, and compound 2 and the positive control Dxd were injected intracavitarially into the tail vein at two doses, 0.3 mg / kg and 1.0 mg / kg, for seven consecutive days. During the adaptation period, cage-side observations were performed once daily, and during the administration period, clinical observations were performed once in the morning and once in the afternoon for all animals. Responses were recorded in various parts of the body, including skin, fur, eyes (sclera), ears, nose, oral cavity, chest, abdomen, urogenital area, and limbs, as well as in aspects such as respiration, movement, urination, defecation (including urine and fecal color), and behavioral changes. Daily weight measurements were recorded for the animals in the main test group, and for animals scheduled for necropsy, final body weight (after fasting) was measured before necropsy. Food intake of the test animals was also recorded. See Table 5-1 for specific test results.

[0242] [Table 7]

[0243] Here, ♂ represents a male and ♀ represents a female.

[0244] In macroscopic autopsies performed on day 8 after administration, localized / punctate dark red discoloration was observed in the lungs of some deceased rats in the Dxd control group, but this lesion was not observed in any of the compound 2-administered rats.

[0245] The experimental data described above demonstrate that compound 2 of this disclosure has lower in vivo toxicity and superior in vivo safety compared to the positive control Dxd.

[0246] In this disclosure, the control compound Dxd refers to the following compound that can be prepared based on the disclosed document WO2014057687A.

[0247] [ka]

Claims

1. A compound of formula II, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. 【Chemistry 1】 (However, R １ H, halogen, OH, SH, NH ２ , C １－４ Alkyl, C １－４ Haloalkyl or C １－４ Selected from alkoxy, R ２ H, halogen, C １－４ Alkyl or C １－４ Selected from alkoxy, Alternatively, R １ and R ２ are cyclized to -O-(CH ２ ), ｍ -O-, where m is selected from 1, 2 or 3, R ３ and R ４ These are H and C, respectively, independently. １－４ Alkyl, C １－４ Alkoxy, C １－４ Haloalkyl or C １－４ Selected from haloalkoxys, Or, R ３ and R ４ is, -(CH ２ ) ｋ - is cyclically formed, where k is selected from 1, 2, 3 or 4, X is H, OH, HO-CH(R ５ )-(CH ２ ) ｐ -CO-NH- or -N(R ６ ) (Caution ７ ) is selected from, where p is selected from 0, 1 or 2, R ５ H, C １－４ Alkyl, C １－４ Selected from haloalkyl, 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl, R ６ H, C １－４ Alkyl or C １－４ Selected from haloalkyl groups, R ７ H or R ８ -S(O) ２ - Selected from, R ８ C １－４ Selected from alkyl, and t is selected from 0, 1, 2, 3, 4, or 5. Furthermore, the compounds of formula II described above do not include the following compounds. 【Chemistry 2】

2. R １ C １－４ It is alkyl and R ２ It is a halogen, Or, R １ is methyl and R ２ F is, Or, R １ is H and R ２ H is, Or, R １ and R ２ is -O-CH ２ Cycloforming into -O-, Or, R １ NH ２ And R ２ is H or halogen, Or, R １ NH ２ And R ２ The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein is H or F.

3. R ３ is H and R ４ H is, Or, R ３ is H and R ４ C １－４ It is alkyl, Or, R ３ is H and R ４ It is methyl, Or, R ３ and R ４ is, -CH ２ -CH ２ A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, which is cyclized to -.

4. X is HO-CH(R ５ )-(CH ２ ) ｐ -CO-NH- and p is selected from 0, 1, and 2. Alternatively, X is OH or NH ２ Selected from, Alternatively, X is H and t is 0. Alternatively, X is -N(R ６ ) (Caution ７ ) a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

5. The compound of formula II is the compound of formula IIa or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, as described in one of claims 1 to 3. 【Transformation 3】 (However, R １ , R ２ , R ３ and R ４ is as defined in any one of claims 1 to 3, and X １ is H, HO-CH(R ５ )-(CH ２ ) ｐ -CO- is selected, p is selected from 0, 1, 2, R ５ H, C １－４ Selected from alkyl, 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl.

6. R １ is selected from methyl or methoxy, or R １ The compound according to claim 5, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein is methyl.

7. R ２ It is selected from F or Cl, or R ２ The compound according to claim 5 or 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein F is F.

8. R １ is methyl and R ２ The compound according to claim 5, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein F is F.

9. R ３ and R ４ is cyclized to -(CH ２ ) ｋ -, and k is 2, or R ３ is H, and R ４ is H, the compound according to any one of claims 5 to 8, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

10. X １ is HO-CH(R ５ )-(CH ２ ) ｐ -CO-, p is selected from 0 or 1, and R ５ is selected from H, C １－４ alkyl or 3- to 6-membered cycloalkyl, Or, X １ HO-CH(R ５ )-(CH ２ ) ｐ -CO-, where p is selected from 0 or 1, and R ５ is selected from H, methyl, or cyclopropyl, Or, X １ HO-CH(R ５ )-CO- and R ５ is selected from H or 3-6 membered cycloalkyl groups. Or, X １ HO-CH(R ５ )-CO- and R ５ is selected from H or cyclopropyl, Or, X １ HO-CH(R ５ ) - CH ２ -CO- and R ５ C １－４ It is alkyl, Or, X １ HO-CH(R ５ ) - CH ２ -CO- and R ５ It is methyl, Or, X １ H, 【Chemistry 4】 A compound according to any one of claims 5 to 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, selected from among.

11. The compound of formula IIa is selected from the compound of formula IIa-1, the compound of formula IIa-2, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, as described in one of claims 5 to 10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. 【Transformation 5】 (However, R １ , R ２ and X １ (This is as defined in any one of claims 5 to 8 and 10.)

12. The compound of formula II is the compound of formula IIb or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, as described in one of claims 1 to 3. 【Transformation 6】 (However, R １ , R ２ , R ３ and R ４ X is defined as any one of claims 1 to 3, ２ is H, OH, or -N(R ６ ) (Caution ７ ) is selected from, R ６ is H or C １－４ Selected from alkyl groups, R ７ H or R ８ -S(O) ２ - Selected from, R ８ C １－４ It is alkyl, and q is selected from 0, 1, 2, 3, and 4.

13. R １ is selected from methyl and R ２ It is selected from H, Cl, F, or R １ and R ２ is -O-(CH ２ ) ｍ The compound according to claim 12, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein the compound is cyclized to -O- and m is selected from 1 or 2.

14. R ３ and R ４ Both are either H or R ３ is H and R ４ The compound according to claim 12 or 13, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein is methyl.

15. X ２ is selected from H, and q is 0. Or, X ２ is OH or -N(R ６ ) (Caution ７ ) is selected from and q is 0, 1, 2, 3 or 4, Or, X ２ is OH or -N(R ６ ) (Caution ７ ) is selected from, where q is 0, 1, 2, 3 or 4, R ３ and R ４ Both are H, and R １ is selected from methyl and R ２ is selected from H, Cl, or F. Or, R １ and R ２ is -O-(CH ２ ) ｍ It is cyclized to -O-, where m is selected from 1 or 2, Or, X ２ H, OH, NH ２ or N(i-Pr)-S(O) ２ -CH ３ A compound according to any one of claims 12 to 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, selected from among.

16. The compound of formula IIb is the compound of formula IIb-1, the compound of formula IIb-2, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, as described in one of claims 12 to 15, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. 【Transformation 7】 (However, R １ , R ２ , X ２ (and q are as defined in any one of claims 12-13 and 15.)

17. The compound of formula IIa is selected from the following compounds or their pharmaceutically acceptable salts or stereoisomers: 【Transformation 8】 Formula IIb is selected from the following compounds or pharmaceutically acceptable salts thereof or stereoisomers thereof, according to claim 5 or 12, the compound or pharmaceutically acceptable salt thereof or stereoisomer thereof. 【Chemistry 9】

18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable excipient.

19. Use of a compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition according to claim 18, in the manufacture of a pharmaceutical for treating tumors.

20. A method for treating a tumor, comprising administering to an individual as required a compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition according to claim 18.

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