Anti-malarial agents

Abstract

The present disclosure relates to compounds of Formula I, and methods of treating a Plasmodium infection, comprising administering to a subject in need thereof a compound of Formula I, or a pharmaceutically acceptable salt thereof. (I)

Description

Cross-references to related applications

[0001] This application is an international patent application filed on June 14, 2023, with international patent application number PCT / CN2023 / 100293, which is incorporated herein by reference in its entirety. Technical Field

[0002] This disclosure relates to compounds of formula I or a pharmaceutically acceptable salt thereof that can be used to treat Plasmodium infection. More specifically, this disclosure relates to compounds of formula I or a pharmaceutically acceptable salt thereof that can be used to treat malaria. Background Technology

[0003] Malaria is a leading human disease, infecting hundreds of millions of people and killing more than 450,000 each year. The deadliest form of malaria is caused by Plasmodium falciparum. This protozoan parasite causes almost all malaria deaths, with the majority occurring in Africa. Plasmodium falciparum has a complex life cycle that begins with the mosquito vector (Anopheles mosquito), at which point the sporozoite form is injected into the human host during a blood meal. These sporozoites migrate to the liver and invade hepatocytes, where they develop to form thousands of hepatic merozoites. These merozoites enter the bloodstream and invade red blood cells to begin the parasite's asexual reproductive cycle, which causes malaria symptoms. The parasite develops within the protected microenvironment of the red blood cell to form 16–32 merozoites, which, once mature, emerge from the host cell to invade new red blood cells. Some of these parasites differentiate to form gametoblasts, the sexual form of the parasite. These can be ingested by mosquitoes, where male and female gametes are formed, fuse, and differentiate into oocysts on the extracellular matrix of the mosquito's midgut cells. Sporozoites are formed within the oocysts and migrate to the salivary glands upon expulsion to be delivered to the next host during blood-feeding, thus enabling the parasite to persist and survive.

[0004] Other forms of malaria include relapsing malaria caused by Plasmodium vivax, which causes significant morbidity, can cause a virulent form of the disease with some mortality, and is primarily a problem outside of Africa. Plasmodium knowlesi is present in Southeast Asia and is a zoonotic parasite that commonly infects macaques, but has been shown to infect humans in Malaysian Borneo.

[0005] Artemisinin-based combination therapies (ACTs) have become a mainstay of malaria treatment and control. However, due to the growing threat of resistance to ACTs, developing new antimalarial drugs with novel targets that inhibit multiple steps in the parasite's life cycle is an urgent priority in malaria control. These novel antimalarial drugs, as monotherapy or ACT companion drugs, can move towards malaria elimination because they reduce the likelihood of parasites with pre-existing resistance mutations existing in the parasite population.

[0006] Currently, aspartic proteases are major targets for drug development: HIV aspartic proteases have been successfully targeted by clinically used drugs; inhibitors targeting human renin, BACE1, and γ-secretase are under or in clinical development. In the field of antimalarial drugs, Plasmepsin X and IX (PMX and PMIX) from Plasmodium falciparum have been identified as potential targets because inhibitors block the parasite's exit from and invasion of host cells and prevent the maturation of some rod-shaped and micronemal proteins required for this process (Pino P, Caldelari R, Mukheijee B, Vahokoski J, Klages N, Maco B, et al., A multistage antimalarial targets the plasmepsins IX and Xessential for invasion and egress. Science. 2017;358(6362):522-8.). Summary of the Invention

[0007] This invention relates to compounds of formula I: I A, Q, X, and Z are described below.

[0008] This article also describes a method for treating Plasmodium infection, comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier thereof, to a subject in need.

[0009] This article also describes methods for treating malaria, which include administering a compound of formula I or a pharmaceutically acceptable salt thereof to a subject in need.

[0010] This disclosure also provides the use of compositions (including pharmaceutical compositions) for the treatment of malaria, said compositions comprising one or more compounds of this disclosure (e.g., one compound of this disclosure), or tautomers thereof, or pharmaceutically acceptable salts or solvates of said compounds and / or said tautomers, optionally together with one or more additional therapeutic agents, optionally in an acceptable (e.g., pharmaceutically acceptable) carrier or diluent.

[0011] Furthermore, this disclosure provides methods for using pharmaceutical compositions to treat Plasmodium infection, treat malaria, inhibit plasmepsin X, or dually inhibit plasmepsin X and plasmepsin IX, said pharmaceutical compositions comprising one or more compounds in free form or pharmaceutically acceptable salt form, and one or more conventional pharmaceutical excipients. Methods for using combinations of the compounds or salts of this disclosure with one or more additional pharmaceutically active agents are also provided.

[0012] This disclosure also provides methods for inhibiting plasmepsin X, or dual inhibition of plasmepsin X and plasmepsin IX activity, and for treating, preventing, improving and / or delaying the onset of diseases or conditions, wherein inhibition of plasmepsin X and / or plasmepsin IX has or may have a therapeutic effect on said diseases or conditions, such as malaria.

[0013] This disclosure also provides a method for inhibiting the aspartic protease of Plasmodium falciparum. This disclosure further provides a method for blocking the growth of Plasmodium falciparum by inhibiting plasmepsin X. This disclosure also provides a method for blocking the growth of Plasmodium falciparum by inhibiting PMX and PlasmepsinIX.

[0014] This disclosure also provides a method for treating malaria by inhibiting plasmepsin X. This disclosure further provides a method for treating malaria by inhibiting PMX and Plasmepsin IX.

[0015] These and other embodiments of this disclosure are included within the scope of this disclosure, and these embodiments will be described in detail below or will become clear to those skilled in the art.

[0016] The above overview of the technology is non-limiting, and other features and advantages of the technology will become apparent from the following detailed description and claims. Detailed Implementation

[0017] This article describes compounds having structural formula I or pharmaceutically acceptable salts thereof: I Where A is a straight chain or a branched chain, saturated or unsaturated (C3-C) 10 A alkylene group comprising at least one -CH2- group, wherein one or more additional -CH2- groups in A are optionally and independently partially substituted by a subset selected from O, S, NR, CONR, NRCO, SO2, and SO2NR, and wherein one or more hydrogen atoms along A may be independently selected from hydroxyl, halogen, and C. 1-3 Group substitution of haloalkyl groups; X is selected from: (a) Straight or branched, saturated or unsaturated (C3-C) 10 )hydro-ion group, , , , , and (b) (c) (d) (e) (f) (g) Among them, single This indicates the connection point with the nitrogen atom of the tetrahydropyrimidine ring, while the double... Indicates the connection point with Z; J is a six-membered aryl or heteroaryl group selected from phenyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl, wherein the phenyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl are unsubstituted or substituted by one to three groups independently selected from R; G is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, bicyclobutyl, bicyclopentyl, bicyclohexyl, bicycloheptyl, bicyclononyl, pyridyl, pyrimidinyl, benzylpyrimidinyl, pyrazolyl, imidazolyl, and the group is optionally substituted by 1 to 3 R groups; R is hydrogen, halogen, C1-C6 alkylCOOH, COOH, C3-C6 cycloalkyl, C1-C6 alkyl, halo-C1-C6 alkyl, -O halo-C1-C6 alkyl, C1-C6 alkylOH, C1-C6 alkoxy, COC1-C6 alkyl, C1-C6 alkylO-C1-C6 alkyl, heteroaryl or COOC1-C6 alkyl; Z represents a bond, -(CH2). p C(O)(CH2) p -, -phenyl-, -heteroaryl-, wherein the phenyl and heteroaryl groups are optionally substituted with 1 to 3 R groups; Q is selected from: , and (a) (b) (c); Among them, single Indicates the connection point with A, while the double This indicates the connection point with Z.

[0018] R 1 It is hydrogen, halogen, CN, OH, C1-C6 alkoxy, C1-C6 alkyl, C1-C6 alkyl, C1-C6 alkyl COOH, COOH, oxo, COOC1-C6 alkyl, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkyl, -C1-C6 alkyl O halo C1-C6 alkyl, halo C1-C6 alkyl, C1-C6 alkyl OH, CON(R) 2 (R) 3 ), N(R 2 (R) 3 ) or C1-C6 alkyl N(R 2 (R) 3 ); R 2 It is hydrogen, C1-C6 alkylCOOH, COOH, C3-C6 cycloalkyl, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkylOH, COC1-C6 alkyl or COOC1-C6 alkyl; R 3 It is hydrogen, C1-C6 alkylCOOH, COOH, C3-C6 cycloalkyl, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkylOH, COC1-C6 alkyl or COOC1-C6 alkyl; k is an integer from 0 to 4; and p is an integer independently selected from 0 to 4.

[0019] When A is a straight-chain or branched, saturated or unsaturated (C3-C6) hydrocarbon group, the embodiment of Formula 1 is implemented. When A is a straight-chain or branched, saturated or unsaturated C3 hydrocarbon group, a sub-implementation of the aspect of this disclosure is implemented. When A is a straight-chain or branched, saturated or unsaturated C4 hydrocarbon group, a sub-implementation of the aspect of this disclosure is implemented. When A is a straight-chain or branched, saturated or unsaturated C5 hydrocarbon group, a sub-implementation of the aspect of this disclosure is implemented. When A is a straight-chain or branched, saturated or unsaturated C6 hydrocarbon group, a sub-implementation of the aspect of this disclosure is implemented. When A is a straight-chain or branched, saturated or unsaturated (C3-C6) hydrocarbon group, a sub-implementation of the aspect of this disclosure is implemented. 10 In the case of the alkylene group, another sub-implementation is achieved, wherein one or more additional -CH2- groups are optionally and independently partially replaced by a subset selected from O, S, NR, CONR, NRCO, SO2, and SO2NR, and wherein one or more hydrogens along A can be independently selected from hydroxyl, halogen, and C.1-3 The alkyl group is substituted. In another embodiment, A is selected from -(CH2)6-, -(CH2)4-, -(CH2)3- and -(CH2)2-CH=CH-.

[0020] When each R 1 Another embodiment of Formula I is achieved when each R is independently selected from halogens, CN, OH, C1-C6 alkoxy, COOH, oxo, COOC1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkyl, -halogenated C1-C6 alkyl, and C1-C6 alkylOH. 1 Sub-implementations of this aspect of the present disclosure are achieved when each R is independently selected from halogens, OH, C1-C6 alkoxy groups, C1-C6 alkyl groups, -C1-haloC1-C6 alkyl groups, and C1-C6 alkyl OH groups. In yet another sub-implementation, each R 1 Independently selected from OH, methyl, and trifluoromethyl. In one category, R 1 It is OH. In another category, each R 1 Independently selected from OH and methyl. In another category, R 1 It is methyl. In another category, R 1 It is trifluoromethyl.

[0021] When R is selected from hydrogen, halogen, C1-C6 alkylCOOH, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkylOH, and C1-C6 alkoxy, another embodiment of Formula I is realized. When R is selected from hydrogen, CH2COOH, (CH2)2COOH, CH(CH3)COOH, CH3, CH2CH3, (CH2)2OCH3, (CH2)3OCH3, (CH2)2OCH2CH3, (CH2)3OCH2CH3, CH2F, CHF2, CF3, (CH2)2OH, C1-C6 alkyl-O-C1-C6 alkyl, and (CH2)3OH, a sub-embodiment of this aspect of the present disclosure is realized. When R is hydrogen, another sub-embodiment of this aspect of the present disclosure is realized. When R is selected from hydrogen, methyl, CH2COOH, (CH2)2COOH, and CH(CH3)COOH, another sub-embodiment of this aspect of the present disclosure is realized. When R is selected from hydrogen, CH3, CH2CH3, (CH2)2OCH3, (CH2)3OCH3, (CH2)2OCH2CH3, or (CH2)3OCH2CH3, another sub-implementation of this aspect of the present disclosure is implemented. When R is selected from hydrogen, methyl, (CH2)2OCH3, (CH2)3OCH3, (CH2)2OCH2CH3, or (CH2)3OCH2CH3, another sub-implementation of this aspect of the present disclosure is implemented. When R is hydrogen, methyl, or (CH2)2OCH3, another sub-implementation of this aspect of the present disclosure is implemented. When R is hydrogen or (CH2)2OCH3, yet another aspect of the present disclosure is implemented. When R is (CH2)2OCH3, another sub-implementation of this aspect of the present disclosure is implemented. When R is CH2F, CHF2, or CF3, another sub-implementation of this aspect of the present disclosure is implemented.

[0022] When Q is: This implements another embodiment of equation I, where R 1 K, as well as single and double wave lines, are as described in this article.

[0023] When Q is: This implements another embodiment of equation I, where R 1 K, as well as single and double wave lines, are as described in this article.

[0024] When Q is: This implements another embodiment of equation I, where R 1 K, as well as single and double wave lines, are as described in this article.

[0025] When Z is a bond, another implementation of equation I is achieved. When Z is -C(O)(CH2) p Another embodiment of Formula I is achieved when Z is -C(O)-, where p is 0. Yet another embodiment of Formula I is achieved when Z is -C(O)(CH2)3-. Yet another embodiment of Formula (I) is achieved when Z is -C(O)(CH2)3-. Yet another embodiment of Formula (I) is achieved when Z is -C(O)(CH2)-. Yet another embodiment of Formula (I) is achieved when Z is -Phenyl- (optionally substituted with 1 to 3 R groups). Yet another embodiment of Formula I is achieved when Z is -Heteroaryl- (optionally substituted with 1 to 3 R groups).

[0026] When X is a straight chain or a branched chain, saturated or unsaturated (C3-C) 10 When X is a (C3-C6) alkylene group, another embodiment of Formula I is realized. When X is a straight-chain or branched, saturated or unsaturated (C3-C4) alkylene group, a sub-implementation of this aspect is realized. When X is a straight-chain or branched, saturated or unsaturated (C3-C4) alkylene group, another sub-implementation of this aspect is realized.

[0027] When X is When J is as described herein, another embodiment of Formula I is realized. A sub-implementation of this aspect of the present disclosure is realized when J is an unsubstituted fused phenyl ring or substituted with one to three independently selected groups from R. Another sub-implementation of this aspect of the present disclosure is realized when J is selected from fused pyridyl, fused pyrimidinyl, fused pyridazinyl, and fused pyrazinyl groups, wherein the pyridyl, pyrimidinyl, pyridazinyl, and pyrazinyl groups are unsubstituted or substituted with one to three independently selected groups from R. Another sub-implementation of this aspect of the present disclosure is realized when J is fused pyridyl, wherein the pyridyl group is unsubstituted or substituted with one to three independently selected groups from R. Another sub-implementation of this aspect of the present disclosure is realized when J is fused pyrimidinyl, wherein the pyrimidinyl group is unsubstituted or substituted with one to three independently selected groups from R. Another sub-implementation of this aspect of the present disclosure is realized when J is fused pyridazinyl, wherein the pyridazinyl group is unsubstituted or substituted with one to three independently selected groups from R. Another sub-implementation of this aspect of the present disclosure is achieved when J is a fused pyrazinyl group, wherein the pyrazinyl group is unsubstituted or substituted by one to three groups independently selected from R.

[0028] When X is At that time, another implementation of Equation I was achieved.

[0029] When X is Another embodiment of Formula I was implemented. Sub-implementations of this aspect of the present disclosure were implemented when G was selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclobutyl, bicyclopentyl, bicyclohexyl, bicycloheptyl, and bicyclononyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclobutyl, bicyclopentyl, bicyclohexyl, bicycloheptyl, and bicyclononyl groups were unsubstituted or substituted with 1 to 3 R groups. Sub-implementations of this aspect of the present disclosure were implemented when G was selected from unsubstituted or substituted cyclopropyl, cyclobutyl, cyclopentyl, cycloheptyl, and bicyclopentyl groups. Another sub-implementation of this aspect of the present disclosure was implemented when G was phenyl, wherein the phenyl group was unsubstituted or substituted with 1 to 3 R groups. Another sub-implementation of this aspect of the present disclosure was implemented when G was pyridyl, wherein the pyridyl group was unsubstituted or substituted with 1 to 3 R groups. Another sub-implementation of this aspect of the present disclosure is achieved when G is a pyrimidinyl group, wherein the pyrimidinyl group is unsubstituted or substituted by 1 to 3 R groups. Another sub-implementation of this aspect of the present disclosure is achieved when G is a benzylpyrimidinyl group, wherein the benzylpyrimidinyl group is unsubstituted or substituted by 1 to 3 R groups. Another sub-implementation of this aspect of the present disclosure is achieved when G is a pyrazolyl group, wherein the pyrazolyl group is unsubstituted or substituted by 1 to 3 R groups. Another sub-implementation of this aspect of the present disclosure is achieved when G is an imidazoleyl group, wherein the imidazoleyl group is unsubstituted or substituted by 1 to 3 R groups. In yet another aspect of the present disclosure, G is cyclopropyl, cyclobutyl, bicyclopentyl, pyridinyl, phenyl, or trifluoromethyl-phenyl.

[0030] When X is (e) represents another implementation of Equation I.

[0031] When X is (f) represents another implementation of Equation I.

[0032] When X is (g) represents another implementation of Equation I.

[0033] This disclosure also relates to compounds of formula II: II Q, X, and n are as defined in this paper.

[0034] When Q is: This implements another embodiment of Equation II, where k is an integer between 0 and 4, and each R 1 Independently selected from halogens, OH, C1-C6 alkoxy groups, C1-C6 alkyl groups, -C1-haloC1-C6 alkyl groups, and C1-C6 alkyl OH groups. In this subclass of embodiments, each R1 Independently selected from OH, C1-C6 alkyl, and halo-C1-C6 alkyl. In further subclasses, each R 1 It is independently selected from OH, methyl and trifluoromethyl.

[0035] When Q is: This implements another embodiment of Equation II, where k is an integer between 0 and 4, and R 1 The radical is selected from hydrogen, halogen, OH, C1-C6 alkoxy, C1-C6 alkyl, -C1-halogenated C1-C6 alkyl, and C1-C6 alkylOH. In one aspect of this embodiment, k is 0.

[0036] When Q is: This implements another embodiment of Equation II, where k is an integer between 0 and 4, and R 1 Selected from hydrogen, halogen, OH, C1-C6 alkoxy, C1-C6 alkyl, -C1-haloC1-C6 alkyl, and C1-C6 alkylOH. In one aspect of this embodiment, k is 0. In another aspect of this embodiment, k is 1 and R 1 It is a hydroxyl group.

[0037] Another implementation of Formula II is achieved when X is a straight or branched, saturated or unsaturated (C3-C6) hydrocarbon group.

[0038] When X is When J is selected from phenyl, pyridyl, or pyrimidinyl, another embodiment of Formula II is achieved, wherein the phenyl, pyridyl, or pyrimidinyl group is unsubstituted or substituted by one to three groups independently selected from R. When J is an unsubstituted or substituted phenyl group, another embodiment of Formula II is achieved. In a specific embodiment of Formula II, J is phenyl. When J is an unsubstituted or substituted pyridyl group, another embodiment of Formula II is achieved. When J is an unsubstituted or substituted pyrimidinyl group, another embodiment of Formula II is achieved.

[0039] When X is At that time, another implementation of Equation II was achieved.

[0040] When X is Another embodiment of Formula II is achieved when G is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclobutyl, bicyclopentyl, phenyl, pyridyl, pyrimidyl, benzylpyrimidyl, pyrazolyl, and imidazolyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclobutyl, bicyclopentyl, phenyl, pyridyl, pyrimidyl, benzylpyrimidyl, pyrazolyl, and imidazolyl groups are unsubstituted or substituted with 1 to 3 R groups. Another sub-embodiment of this aspect of the disclosure is achieved when G is selected from cyclopropyl, cyclobutyl, bicyclobutyl, bicyclopentyl, phenyl, pyridyl, pyrimidyl, and benzylpyrimidyl, wherein the cyclopropyl, cyclobutyl, bicyclobutyl, bicyclopentyl, phenyl, pyridyl, pyrimidyl, and benzylpyrimidyl groups are unsubstituted or substituted with 1 to 3 R groups. When G is selected from unsubstituted or substituted cyclopropyl, another sub-implementation of this aspect of the present disclosure is achieved. When G is selected from unsubstituted or substituted cyclobutyl, another sub-implementation of this aspect of the present disclosure is achieved. When G is selected from unsubstituted or substituted bicyclobutyl, another sub-implementation of this aspect of the present disclosure is achieved. When G is selected from unsubstituted or substituted bicyclopentyl, another sub-implementation of this aspect of the present disclosure is achieved. When G is selected from unsubstituted or substituted phenyl, another sub-implementation of this aspect of the present disclosure is achieved. When G is selected from unsubstituted or substituted pyridyl, another sub-implementation of this aspect of the present disclosure is achieved. When G is selected from unsubstituted or substituted benzylpyrimidinyl, another sub-implementation of this aspect of the present disclosure is achieved.

[0041] When X is (e) represents another implementation of Equation II.

[0042] When X is (f) implements another embodiment of equation II.

[0043] When X is (g) represents another implementation of Equation II.

[0044] Another embodiment of Formula II is achieved when R is selected from hydrogen, halogen, C1-C6 alkylCOOH, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkylOH, and C1-C6 alkoxy. Another embodiment of Formula II is achieved when R is selected from hydrogen, CH2COOH, (CH2)2COOH, CH(CH3)COOH, CH3, CH2CH3, (CH2)2OCH3, (CH2)3OCH3, (CH2)2OCH2CH3, (CH2)3OCH2CH3, CH2F, CHF2, CF3, -OCH2F, -OCHF2, -OCF3, (CH2)2OH, (CH2)3OH, and pyridyl. In one aspect of this embodiment of Formula II, R is selected from hydrogen, methyl, and (CH2)2OCH3.

[0045] When n is 0, another implementation of Equation II is achieved. When n is 1, another implementation of Equation II is achieved. When n is 2, another implementation of Equation II is achieved. When n is 3, another implementation of Equation II is achieved.

[0046] In each of the various embodiments of this disclosure, and in the compounds used in the methods herein, it should be understood that, for each variable (including each of Formulas I-II and those in their various embodiments), each variable is chosen independently of the others unless otherwise stated.

[0047] In each of the various embodiments of this disclosure, the compounds described herein, including each of formulas I-II and those in their various embodiments, may be present in different forms of the compounds, such as any solvates, hydrates, stereoisomers and tautomers of the compounds and any pharmaceutically acceptable salts thereof.

[0048] Definitions and abbreviations : The terms used herein have their general meanings, and the meaning of these terms is independent each time they appear. Nevertheless, and unless otherwise stated, the following definitions apply throughout the specification and claims. Chemical names, common names, and chemical structures are used interchangeably to describe the same structure. Unless otherwise stated, these definitions apply whether a term is used alone or in combination with other terms. Thus, the definition of "alkyl" applies to "alkyl" as well as the "alkyl" portion of "hydroxyalkyl," "haloalkyl," arylalkyl, alkylaryl, "alkoxy," etc.

[0049] It should be understood that in the various embodiments of the disclosure described herein, any variables not explicitly defined in the context of the implementation are as defined in Equation I.

[0050] In the various implementations described herein, each variable is chosen independently of the others unless otherwise stated.

[0051] "Drug resistance associated with Plasmodium strains" refers to Plasmodium species that are no longer susceptible to at least one previously effective drug; they have developed the ability to tolerate attacks by at least one previously effective drug. Drug-resistant strains can pass on this tolerance to their progeny. This resistance may be due to random genetic mutations in the bacterial cells that alter their sensitivity to a single drug or to different drugs.

[0052] "Patients" include both humans and non-human animals. Non-human animals include research animals and companion animals such as mice, rats, primates, monkeys, chimpanzees, great apes, dogs, and domestic cats.

[0053] A “pharmaceutical composition” (or “pharmaceuticalally acceptable composition”) means a composition suitable for administration to a patient. Such compositions may contain a net compound (or multiple compounds) of this disclosure or a mixture thereof, or a salt, solvate, prodrug, isomer, or tautomer thereof, and one or more pharmaceutically acceptable carriers or diluents. The term “pharmaceutical composition” is also intended to cover a bulk composition and individual dose units consisting of one or more (e.g., two) pharmaceutically active agents (e.g., compounds of this disclosure and other agents selected from a separate list of agents described herein) and any pharmaceutically inactive excipients. The bulk composition and each individual dose unit may contain a fixed amount of the aforementioned “more than one pharmaceutically active agent”. The bulk composition is material not yet formed into individual dose units. An illustrative dose unit is an oral dose unit, such as a tablet, pill, etc. Similarly, the methods of treating a patient by administering a pharmaceutical composition of this disclosure described herein are also intended to include administering the aforementioned bulk composition and individual dose units.

[0054] "Halogen" and "halogenated" refer to fluorine, chlorine, bromine, or iodine. Fluorine, chlorine, and bromine are preferred.

[0055] "Hydroalkylene group," either itself or as a substituent, refers to a divalent hydrocarbon chain group having the stated number of carbon atoms. For example, -(C1-C5)hydroalkylene groups include, for example, -CH2-, -CH2CH2-, -CH2CH2CH2CH2-, -CH2CH(CH3)CH2-, or -CH2CH2CH2CH2CH2-. A straight-chain hydroalkylene group refers to a divalent straight-chain hydrocarbon chain group having the stated number of carbon atoms. A branched hydroalkylene group refers to a divalent branched hydrocarbon chain group having the stated number of carbon atoms. A saturated hydroalkylene group refers to a divalent saturated hydrocarbon chain group having the stated number of carbon atoms. An unsaturated hydroalkylene group refers to a divalent hydrocarbon chain group having the stated number of carbon atoms and one or more covalent double or triple bonds within the chain. A cyclic hydroalkylene group refers to a divalent hydrocarbon chain group having the stated number of carbon atoms and a cycloalkyl portion within the chain.

[0056] "Alkyl" refers to an aliphatic hydrocarbon group, which may be straight-chain or branched and contains about 1 to about 20 carbon atoms in the chain. Preferred alkyl groups contain about 1 to about 12 carbon atoms in the chain. More preferred alkyl groups contain about 1 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups (such as methyl, ethyl, or propyl) are attached to a straight-chain alkyl chain. "Lower alkyl" refers to a group having about 1 to about 6 carbon atoms in the chain, which may be straight-chain or branched. Non-limiting examples of suitable alkyl groups include methyl, ethyl, n-propyl, isopropyl, and tert-butyl.

[0057] "Halogenated alkyl" refers to an alkyl group as defined above, wherein one or more hydrogen atoms on the alkyl group are replaced by a halogen group as defined above.

[0058] "Aryl" refers to an aromatic monocyclic or polycyclic ring system containing about 6 to about 14 carbon atoms, preferably about 6 to about 10 carbon atoms. The aryl group may optionally be substituted by one or more "cyclic substituents," which may be the same or different and are defined herein as "cyclic substituents." Non-limiting examples of suitable aryl groups include phenyl and naphthyl. "Monocyclic aryl" refers to phenyl.

[0059] “Cycloalkyl” refers to a non-aromatic monocyclic or polycyclic ring system comprising about 3 to about 12 carbon atoms, preferably about 3 to about 10 carbon atoms. Preferred cycloalkyl rings comprise about 5 to about 10 ring atoms. The cycloalkyl group may optionally be substituted with one or more substituents, which may be the same or different, as described herein. Monocyclic cycloalkyl refers to the monocyclic form of the cycloalkyl moiety described herein. Non-limiting examples of suitable monocyclic cycloalkyl groups include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, etc. Polycyclic cycloalkyl refers to a polycyclic (including bicyclic) ring comprising a non-aromatic ring. Non-limiting examples of suitable polycyclic cycloalkyl groups include 1-decahydronaphthyl, norbornyl, adamantyl, etc. In some embodiments, the non-aromatic ring is fused with an aromatic ring.

[0060] The term "heteroaryl" refers to an aromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system having 1-3 heteroatoms for the monocyclic, 1-6 heteroatoms for the bicyclic, or 1-9 heteroatoms for the tricyclic, wherein the heteroatoms are selected from O, N, or S (e.g., a carbon atom and 1-3, 1-6, or 1-9 N, O, or S heteroatoms for the monocyclic, bicyclic, or tricyclic rings, respectively). Non-limiting examples of heteroaryls are pyridyl, pyrazolyl, pyrimidinyl, furanyl, oxazolyl, triazolyl, oxadiazolyl, and thiophenyl. Heteroaryls described herein may also contain fused rings sharing a common carbon-carbon bond.

[0061] "Heterocyclic alkyl" (or "heterocyclic group") refers to a non-aromatic, saturated or partially saturated monocyclic or polycyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, wherein one or more atoms in the ring system are elements other than carbon, such as nitrogen, oxygen or sulfur alone or in combination. Adjacent oxygen and / or sulfur atoms are not present in the ring system. Preferred heterocyclic groups contain about 5 to about 6 ring atoms. The prefixes nitro, oxo, or thio before the name of the heterocyclic group indicate the presence of at least one nitrogen, oxygen, or sulfur atom as a ring atom, respectively. Any -NH in the heterocyclic group ring may be protected, such as, for example, -N(Boc), -N(CBz), -N(Tos) groups, etc.; such protection is also considered part of this disclosure. The heterocyclic group may optionally be substituted by one or more substituents, which may be the same or different, as described herein. The nitrogen or sulfur atom of the heterocyclic group may optionally be oxidized to the corresponding N-oxide, S-oxide, or S,S-dioxide. Therefore, when the term "oxide" appears in the definition of variables in the general structure described herein, it refers to the corresponding N-oxide, S-oxide, or S,S-dioxide. "Heterocyclic group" also includes a ring in which the =O group replaces two available hydrogen atoms on the same carbon atom (i.e., a heterocyclic group includes a ring having a carbonyl group in the ring). Such =O groups may be referred to herein as "oxo-substituted". An example of such a part is pyrrolidone (or pyrrolidone): As used herein, the term "monocyclic heterocyclic alkyl" refers to the monocyclic form of the heterocyclic alkyl moiety described herein, and includes 4- to 7-membered monocyclic heterocyclic alkyl groups comprising 1 to 4 cyclic heteroatoms independently selected from N, N-oxide, O, S, S-oxide, S(O), and S(O)2. The junction with the parent moiety is any available cyclic carbon or cyclic heteroatom. Non-limiting examples of monocyclic heterocyclic alkyl groups include piperidinyl, oxetyl, pyrrolyl, piperazine, morpholinyl, thiomorpholinyl, thiazolyl, 1,4-dioxyl, tetrahydrofuranyl, tetrahydrothiophenyl, β-lactam, γ-lactam, δ-lactam, β-lactone, γ-lactone, δ-lactone, and pyrrolidone and their oxides. Non-limiting examples of monocyclic heterocyclic alkyl groups include: Non-limiting examples of polycyclic heterocyclic alkyl groups include bicyclic heterocyclic alkyl groups. Specific examples include, but are not limited to, those that... and .

[0062] "Alkoxy" refers to an alkyl-O- group, wherein the alkyl group is as described above. Non-limiting examples of suitable alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, and n-butoxy. The bond to the parent compound is via an ether oxygen.

[0063] The term "substituted" means that one or more hydrogen atoms on a specified atom are replaced by a selective group of a specified group, provided that the substitution does not exceed the normal valence of the specified atom under its existing conditions, and that the substitution produces a stable compound. Such a combination is permitted only if the combination of substituents and / or variables produces a stable compound. A "stable compound" or "stable structure" means a compound that is robust enough to withstand separation from the reaction mixture to a useful purity and formulation into an effective therapeutic agent.

[0064] The term "optionally substituted" refers to the optional substitution of a specified group, radical, or part.

[0065] When a variable appears more than once in a group, such as R8 in -N(R8)2, or when a variable appears more than once in the structure presented herein, the variable may be the same or different each time it appears.

[0066] solid line As a bond, it usually represents a mixture or either of the possible isomers, such as those containing (R)- and (S)- stereochemistry. For example: Indicates inclusion and One or two of them.

[0067] As shown in this article, the wavy lines cross the lines representing chemical bonds. Indicates the connection point with the rest of the compound. Draw lines into the ring system, such as, for example... This indicates that the indicated line (bond) can be connected to any substituted ring atom.

[0068] "Oxygenation" is defined as an oxygen atom double-bonded to a cyclic carbon atom in a cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, or another ring as described herein, for example... .

[0069] In this specification, when multiple oxygen and / or sulfur atoms are present in a ring system, no adjacent oxygen and / or sulfur atoms may be present in the ring system.

[0070] As is well known in the art, unless otherwise stated, a bond drawn from a particular atom (without depicting the portion at the end of the bond) represents a methyl group bonded to that atom via that bond. For example: express .

[0071] In another embodiment, compounds that can be used in the methods of this disclosure and / or compositions comprising them that can be used in the methods are present in an isolated and / or purified form. For compounds, the terms “purified,” “purified form,” or “isolated and purified form” refer to the physical state of the compound after isolation from a synthetic process (e.g., from a reaction mixture) or a natural source or combination thereof. Thus, for compounds, the terms “purified,” “purified form,” or “isolated and purified form” refer to the physical state of the compound (or its tautomers or stereoisomers, or pharmaceutically acceptable salts or solvates of the compound, the stereoisomers, or the tautomers) obtained from one or more purification methods (e.g., chromatography, recrystallization, etc.) described herein or known to those skilled in the art, having a purity sufficient for in vivo or pharmaceutical use and / or characterizable by standard analytical techniques described herein or known to those skilled in the art.

[0072] It should be understood that any carbon and heteroatom with an unsatisfied valence in the text, schemes, embodiments and tables herein are assumed to have a sufficient number of hydrogen atoms to satisfy the valence.

[0073] When a functional group in a compound is referred to as “protected,” it means that when the compound reacts, the group is in a modified form to exclude undesirable side reactions at the protected site. Suitable protecting groups are those recognized by those skilled in the art and by reference to standard textbooks (e.g., TW Greene et al., Bioorg. Med. Chem. Lett. (1998); Protective Groups in Organic Synthesis (1991), Wiley, New York).

[0074] Another embodiment provides a prodrug and / or solvate of the compounds disclosed herein. A discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) 14, ACS Symposium Series, and Bioreversible Carriers in Drug Design, (1987), edited by Edward B. Roche, American Pharmaceutical Association and Pergamon Press. The term “prodrug” means a compound (e.g., a drug precursor) that is converted in vivo to produce the compounds of this disclosure or pharmaceutically acceptable salts, hydrates, or solvates of such compounds. Conversion can occur through various mechanisms (e.g., via metabolic or chemical processes), such as, for example, via hydrolysis in the blood. A discussion of the use of prodrugs is provided in T. Higuchi and W. Stella, “Pro-drugs as Novel Delivery Systems,” ACS Symposium Series, Volume 14, and Bioreversible Carriers in Drug Design, edited by Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.

[0075] For example, if the compound or its pharmaceutically acceptable salt that can be used in the methods of this disclosure contains a carboxylic acid functional group, the prodrug may comprise an ester formed by replacing the hydrogen atom of the acid group with the following groups: such as (C1-C8) alkyl, (C2-C12) alkoxymethyl, 1-(alkoxy)ethyl having 4-9 carbon atoms, 1-methyl-1-(alkoxy)-ethyl having 5-10 carbon atoms, or alkoxycarbonyloxymethyl having 3-6 carbon atoms. 1-(alkoxycarbonyloxy)ethyl with 4-7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl with 5-8 carbon atoms, N-(alkoxycarbonyl)aminomethyl with 3-9 carbon atoms, 1-(N-(alkoxycarbonyl)amino)ethyl with 4-10 carbon atoms, 3-phthaloyl, 4-crotonic acid lactone, γ-butyrolactone-4-yl, di-N,N-(C1-C2)alkylamino(C2-C3)alkyl (such as β-dimethylaminoethyl), carbamoyl-(C1-C2)alkyl, N,N-di(C1-C2)alkylcarbamoyl-(C1-C2)alkyl and piperidinyl-, pyrrolidine- or morpholino(C2-C3)alkyl, etc.

[0076] Similarly, if the compound used in the methods of this disclosure contains an alcohol functional group, it can be formed, for example, by replacing the hydrogen atom of the alcohol group with a group such as (C1-C6)alkoxymethyl, 1-((C1-C6)alkoxy)ethyl, 1-methyl-1-((C1-C6)alkoxy)ethyl, (C1-C6)alkoxycarbonyloxymethyl, N-(C1-C6)alkoxycarbonylaminomethyl, succinyl, (C1-C6)alkoxy, α-amino(C1-C4)alkyl, aryl acyl, and α-aminoacyl or α-aminoacyl-α-aminoacyl, wherein each α-aminoacyl is independently selected from naturally occurring L-amino acids, P(O)(OH)2, -P(O)(O(C1-C6)alkyl)2, or glycosyl (a group generated by removing the hydroxyl group of a carbohydrate in the form of a hemiacetal), etc.

[0077] If the compound used in the methods of this disclosure contains an amine functional group, the prodrug can be formed by replacing the hydrogen atom in the amine group with the following groups: such as, for example, R-carbonyl, RO-carbonyl, NRR′-carbonyl, wherein R and R′ are each independently (C1-C10)alkyl, (C3-C7)cycloalkyl, benzyl, or R-carbonyl is a native α-aminoacyl or native α-aminoacyl, -C(OH)C(O)OY1, wherein Y1 is H, (C1-C 6) Alkyl or benzyl, -C(OY2)Y3, wherein Y2 is (C1-C4)alkyl and Y3 is (C1-C6)alkyl, carboxyl (C1-C6)alkyl, amino (C1-C4)alkyl or mono-N- or di-N,N-(C1-C6)alkylaminoalkyl, -C(Y4)Y5, wherein Y4 is H or methyl and Y5 is mono-N- or di-N,N-(C1-C6)alkylaminomorpholinyl, piperidin-1-yl or pyrrolidine-1-yl, etc.

[0078] One or more compounds used in the methods of this disclosure may be present in a non-solventized form and in a solvated form with pharmaceutically acceptable solvents such as water, ethanol, etc., and this disclosure is intended to include both solvated and non-solventized forms. "Solvate" means the physical association of a compound of this disclosure with one or more solvent molecules. Such physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In some cases, solvates can be separated, for example when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. "Solvate" includes solution phases and separable solvates. Non-limiting examples of suitable solvates include ethanolides, methanolides, etc. "Hydrate" is a solvate in which the solvent molecule is H₂O.

[0079] One or more compounds used in the methods of this disclosure may optionally be converted into solvates. The preparation of solvates is generally known. Thus, for example, the preparation of antifungal fluconazole in ethyl acetate and from water is described, as well as in J. Pharmaceutical Sci., 1990, 3, 601-611. Similar preparations of solvates, semisolvents, hydrates, etc., are described in EC van Tonder et al., AAPS Pharm Sci Tech., 5(1), article 12 (2004); and AL Bingham et al., Chem. Commun., 603-604 (2001). Typical non-limiting methods involve dissolving the compound of the invention in a desired amount of a desired solvent (organic or water or a mixture thereof) at a temperature above ambient temperature, cooling the solution at a rate sufficient to form crystals, and then separating the crystals by standard methods. Analytical techniques, such as, for example, IR spectroscopy, show the presence of the solvent (or water) in the crystals as solvates (or hydrates).

[0080] "Effective amount" or "therapeutic effective amount" means the amount by which a compound or composition used in the methods described herein effectively inhibits the aforementioned disease or enzyme activity and thus produces the desired therapeutic, ameliorative, inhibitory, or preventative effect.

[0081] Another embodiment provides pharmaceutically acceptable salts of compounds used in the methods of this disclosure. Therefore, unless otherwise stated, reference to a compound used in the methods disclosed herein should be understood to include reference to its salt. As used herein, the term "salt" means an acidic salt formed with inorganic and / or organic acids, and a basic salt formed with inorganic and / or organic bases. Furthermore, when the compounds of this disclosure contain a basic moiety (e.g., but not limited to pyridine or imidazole) and an acidic moiety (e.g., but not limited to carboxylic acids), zwitterions ("internal salts") can be formed and included within the term "salt" as used herein. Pharmaceutically acceptable (i.e., non-toxic and physiologically acceptable) salts are preferred, although other salts are also useful. Salts of compounds used in the methods of this disclosure can be formed, for example, by reacting the compounds of this disclosure with a certain amount (e.g., an equivalent amount) of an acid or base in a medium (e.g., a medium in which the salt precipitates) or in an aqueous medium, followed by lyophilization.

[0082] Exemplary acid addition salts include acetates, ascorbic acid salts, benzoates, benzenesulfonates, hydrogen sulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, fumarates, hydrochlorides, hydrobroms, hydroiodates, lactates, maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propionates, salicylates, succinates, sulfates, tartrates, thiocyanates, toluenesulfonates (also known as p-toluenesulfonates), etc.

[0083] Additionally, for example, P. Stahl et al., Camille G. (eds.), Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002), Zurich: Wiley-VCH; S. Berge et al., Journal of Pharmaceutical Science (1977), 66(1), 1-19; P. Gould, International J. of Pharmaceutics (1986), 33, 201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; and The Orange Book (Food & Drug Administration, Washington, DC, on its website) discusses acids generally considered suitable for forming pharmaceutically useful salts from basic pharmaceutical compounds. These publications are incorporated herein by reference.

[0084] Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (e.g., organic amines) such as dicyclohexylamine and tert-butylamine, and salts with amino acids such as arginine and lysine. The basic nitrogen-containing group can be quaternized with reagents such as lower alkyl halides (e.g., methyl, ethyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (e.g., dimethyl sulfate, diethyl sulfate, and dibutyl sulfate), long-chain halides (e.g., decyl, lauryl, and stearyl chlorides, bromides, and iodides), and aralkyl halides (e.g., benzyl and phenethyl bromides).

[0085] All of these acid salts and base salts are intended to be pharmaceutically acceptable salts within the scope of this disclosure, and for the purposes of this disclosure, all acid salts and base salts are considered equivalent to the free form of the corresponding compound.

[0086] Another embodiment provides pharmaceutically acceptable esters for use in the methods of this disclosure. Such esters include the group consisting of: (1) carboxylic acid esters obtained by esterification of a hydroxyl group, wherein the non-carbonyl portion of the carboxylic acid moiety of the ester group is selected from straight-chain or branched alkyl groups (e.g., acetyl, n-propyl, tert-butyl, or n-butyl), alkoxyalkyl groups (e.g., methoxymethyl), aralkyl groups (e.g., benzyl), aryloxyalkyl groups (e.g., phenoxymethyl), aryl groups (e.g., phenyl groups optionally substituted with, for example, halogens, C1-4 alkyl groups, or C1-4 alkoxy groups or amino groups); (2) sulfonates, such as alkylsulfonyl or aralkylsulfonyl groups (e.g., methanesulfonyl); (3) amino acid esters (e.g., L-valine or L-isoleucyl); (4) phosphonates; and (5) monophosphates, diphosphates, or triphosphates. Phosphate esters may be further esterified, for example, by C1-20 alcohols or reactive derivatives thereof, or by 2,3-bis(C6-24)acylglycerols.

[0087] As described herein, another embodiment provides tautomers of the compounds of this disclosure for use in the methods herein, as well as salts, solvates, esters, and prodrugs of said tautomers. It should be understood that all tautomer forms of these compounds are within the scope of the compounds used in the methods of this disclosure. For example, all keto-enol and imine-enamine forms of the compounds are included in this disclosure, when present.

[0088] Compounds used in the methods of this disclosure may contain asymmetric or chiral centers and therefore exist in different stereoisomeric forms. It is intended that all stereoisomers of the compounds used in the methods of this disclosure, as well as mixtures thereof (including racemic mixtures), constitute part of this disclosure. Furthermore, this disclosure includes the use of all geometric and positional isomers. For example, if the compounds used in the methods of this disclosure contain double bonds or fused rings, then the cis and trans forms, (E) and (Z) forms, and mixtures thereof are included within the scope of this disclosure.

[0089] Another embodiment provides diastereomeric mixtures and individual enantiomers of compounds used in the methods of this disclosure. The diastereomeric mixture can be separated into its individual enantiomers based on their physicochemical differences using methods well known to those skilled in the art (e.g., by chromatography and / or fractional crystallization). Enantiomers can be separated by reacting the enantiomer mixture with a suitable optically active compound (e.g., a chiral auxiliary agent, such as a chiral alcohol or Mosher acyl chloride), converting the enantiomer mixture into a diastereomeric mixture, separating the diastereomeric isomers, and converting (e.g., hydrolyzing) individual diastereomeric isomers into their respective pure enantiomers. Furthermore, some compounds used in the methods of this disclosure can be transisomers (e.g., substituted biaryl groups) and are considered part of this disclosure. Enantiomers can also be separated using a chiral HPLC column.

[0090] All stereoisomers of the compounds used in the methods of this disclosure (including those stereoisomers of salts, solvates, esters, and prodrugs of the compounds, as well as those stereoisomers of salts, solvates, and esters of prodrugs) (e.g., geometric isomers, optical isomers, etc.), such as those that may exist due to asymmetric carbons on various substituents, including enantiomers (which can even exist in the absence of asymmetric carbons), rotational isomers, trans-blocking isomers, and diastereomeric forms, are considered embodiments within the scope of this disclosure, as are positional isomers (e.g., 4-pyridyl and 3-pyridyl). (For example, if the compounds of this disclosure contain double bonds or fused rings, the cis and trans forms, as well as mixtures thereof, are included within the scope of this disclosure. Furthermore, for example, all keto-enol and imine-enamine forms of the compounds are included in the methods of this disclosure.)

[0091] Individual stereoisomers of the compounds disclosed herein may, for example, be substantially free of other isomers, or may be, for example, as racemates or mixed with all other stereoisomers or other alternative stereoisomers. The chiral center of the disclosed compound may have an S or R configuration as recommended by IUPAC 1974. The terms “salt,” “solvent,” “ester,” “prodrug,” etc., are intended to equally apply to salts, solvates, esters, and prodrugs of enantiomers, stereoisomers, rotational isomers, tautomers, positional isomers, racemates, or prodrugs of the compounds of this invention.

[0092] Another embodiment provides compounds for isotopic labeling in the methods of the present invention. Such compounds are the same as those described herein, but in that one or more atoms are replaced by atoms having atomic masses or mass numbers different from those commonly found in nature. Examples of isotopes that can be incorporated into the compounds of this disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2H, 3H, 13C, 14C, 15N, 18O, 17O, 31P, 32P, 35S, 18F, and 36Cl, respectively.

[0093] Certain isotopically labeled compounds of this disclosure (e.g., those labeled with 3H and 14C) can be used for the determination of compound and / or substrate tissue distribution. Tritium (i.e., 3H) and carbon-14 (i.e., 14C) isotopes are particularly preferred due to their ease of preparation and detectability. Furthermore, substitution with heavier isotopes such as deuterium (i.e., 2H) can provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dose requirement), and may therefore be preferred in some cases.

[0094] The isotopically labeled compounds disclosed herein can generally be prepared by replacing non-isotopically labeled reagents with appropriate isotopically labeled reagents by methods similar to those disclosed in the schemes and / or examples below.

[0095] In the compounds used in the methods of this disclosure, atoms may exhibit their natural isotopic abundance, or one or more atoms may be artificially enriched with specific isotopes having the same atomic number but different atomic mass or mass number from those predominantly found in nature. This disclosure is intended to include all suitable isotopic variations of the compounds of this disclosure. For example, different isotopic forms of hydrogen (H) include protium (¹H) and deuterium (²H). The presence of deuterium in the compounds of this disclosure is indicated by “D”. Protium is the predominant hydrogen isotope found in nature. Enrichment of deuterium may provide certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or may provide compounds that can be used as standards for characterizing biological samples. The isotopically enriched compounds of this disclosure can be prepared using conventional techniques well known to those skilled in the art or by methods similar to those described in the schemes and examples herein, using appropriate isotopic enrichment reagents and / or intermediates without excessive experimentation.

[0096] The polymorphic forms of the compounds used in the methods of this disclosure, as well as the polymorphic forms of the salts, solvates, esters, and prodrugs of the compounds of this disclosure, are intended to be included in this disclosure.

[0097] Treatment This disclosure relates to a method for treating Plasmodium infection, comprising administering to a subject in need a compound described herein or a pharmaceutically acceptable salt thereof. More specifically, the method of this disclosure comprises administering a compound of formula I or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered in the form of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier or excipient.

[0098] This disclosure provides a method for treating Plasmodium infection or for treating malaria or for inhibiting plasmepsin X, the method comprising administering to a subject in need a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof, said compound having structural formula I as described in the summary of the invention of this disclosure. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered as a pharmaceutical composition together with a pharmaceutically acceptable carrier. Various embodiments of these methods are also provided herein, as described below.

[0099] This disclosure also relates to the use of compounds of formula I-II or pharmaceutically acceptable salts thereof for inhibiting plasmepsin X activity, for treating Plasmodium infection, or for treating malaria. This disclosure also relates to the use of compounds of formula I-II or pharmaceutically acceptable salts thereof in the preparation of medicaments for inhibiting plasmepsin X activity, for treating Plasmodium infection, or for treating malaria. Compounds of formula I-II or pharmaceutically acceptable salts thereof described in any of the embodiments disclosed herein may be used for any of the above uses.

[0100] This disclosure provides a method for treating Plasmodium infection, for treating malaria, or for inhibiting plasmepsin IX, the method comprising administering to a patient requiring such treatment a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof, said compound having structural formula I as described in the summary of the invention of this disclosure. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered as a pharmaceutical composition together with a pharmaceutically acceptable carrier. Various embodiments of these methods are also provided herein, as described below.

[0101] This disclosure also relates to the use of compounds of formula I-II or pharmaceutically acceptable salts thereof for inhibiting plasmepsin IX activity, for treating Plasmodium infection, or for treating malaria. This disclosure also relates to the use of compounds of formula I-II or pharmaceutically acceptable salts thereof in the preparation of medicaments for inhibiting plasmepsin IX activity, for treating Plasmodium infection, or for treating malaria. Compounds of formula I-II or pharmaceutically acceptable salts thereof described in any of the embodiments disclosed herein may be used for any of the above uses.

[0102] This disclosure provides a method for treating Plasmodium infection or malaria, or for inhibiting plasmepsin X and plasmepsin IX, the method comprising administering to a subject requiring such treatment a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof, the compound being of structural formula I as described in the summary of this disclosure. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered as a pharmaceutical composition together with a pharmaceutically acceptable carrier. Various embodiments of these methods are also provided herein, as described below.

[0103] This disclosure also relates to the use of compounds of formula I-II or pharmaceutically acceptable salts thereof for inhibiting the activity of plasmepsin X and plasmepsin IX, for treating Plasmodium infection, or for treating malaria. This disclosure also relates to the use of compounds of formula I-II or pharmaceutically acceptable salts thereof in the preparation of medicaments for inhibiting the activity of plasmepsin X and plasmepsin IX, for treating Plasmodium infection, or for treating malaria. The compounds of formula I-II or pharmaceutically acceptable salts thereof described in any of the embodiments disclosed herein may be used for any of the above uses.

[0104] The methods disclosed herein can be used to treat malaria because they suppress the onset, growth, or progression of the disease, improve the symptoms of the disease, induce remission of the disease, cure the disease, or otherwise improve the overall health of subjects who have the disease or are at risk of developing the disease. Therefore, in accordance with the subject matter disclosed herein, the terms “treat,” “treating,” and their grammatical variations, as well as the phrase “treatment method,” are intended to cover any desired therapeutic intervention, including but not limited to methods for treating existing infections in infected subjects, such as in subjects who have been exposed to parasites as disclosed herein.

[0105] Embodiments of this disclosure also include one or more compounds of formula I-II or pharmaceutically acceptable salts thereof, used for: (ii) as a medicament or composition or (iii) for the preparation of a medicament used for: (a) treatment (e.g., treatment in humans); (b) a pharmaceutical agent; (c) inhibiting the growth of parasites / malaria parasites; (d) treating or preventing infection with malaria parasite species; (e) reducing the progression, onset, or severity of pathological symptoms associated with malaria parasite infection, and / or reducing the likelihood of severe malaria parasite infection; or (f) treating, preventing, or delaying the onset, severity, or progression of malaria parasite-related diseases, including but not limited to: malaria.

[0106] Therefore, another embodiment provides a method for treating malaria or for treating Plasmodium infection, comprising administering a combination of a compound comprising at least one of formulas I-II or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, and an effective amount of one or more additional agents described below. In some embodiments, methods for treating malaria or for treating Plasmodium infection are described herein, comprising administering a combination of a compound comprising at least one of formulas I-II or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, and an effective amount of one or more additional antimalarial agents. In some embodiments, methods for treating malaria by inhibiting plasmepsin X, IX, and at least one other mechanism are described herein, comprising administering a combination of a compound comprising at least one of formulas I-II or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, and an effective amount of one or more additional antimalarial agents, wherein the additional antimalarial agents act through a mechanism different from inhibiting plasmepsin IX or plasmepsin X. The pharmacological properties of compounds of formulas I-II or pharmaceutically acceptable salts thereof can be confirmed by several pharmacological assays.

[0107] Dosage and administration Another embodiment provides suitable dosages and dosage forms of the compounds used in the methods of this disclosure. Suitable dosages of the compounds used in the methods of this disclosure for administration to a patient can be readily determined by those skilled in the art, such as by an attending physician, pharmacist, or other technical personnel, and can vary depending on the patient's health, age, weight, frequency of administration, use with other active ingredients, and / or the indication for which the compound is administered. Dosage ranges can be from about 0.001 to 500 mg / kg body weight / day of the compounds of this disclosure. In one embodiment, the dosage is from about 0.01 to about 25 mg / kg body weight / day of the compounds of this disclosure or a pharmaceutically acceptable salt or solvate of said compounds. In another embodiment, depending on the specific application, the amount of active compound in a unit dose formulation can vary or be adjusted from about 1 mg to about 100 mg, in specific embodiments from about 1 mg to about 50 mg, and in specific embodiments from about 1 mg to about 25 mg. In another embodiment, the typical recommended daily dosage regimen for oral administration can range from about 1 mg / day to about 500 mg / day, in specific embodiments from 1 mg / day to 200 mg / day, in 2 to 4 fractional doses.

[0108] As stated above, the dosage and frequency of administration of the disclosed compounds and / or their pharmaceutically acceptable salts are adjusted according to the judgment of the attending physician, taking into account factors such as the patient's age, condition and body size, and the severity of the symptoms being treated.

[0109] Liquid formulations include solutions, suspensions, and emulsions. Examples include water or water-propylene glycol solutions for parenteral injection, or the addition of sweeteners and opacifiers for oral solutions, suspensions, and emulsions. Liquid formulations may also include solutions for intranasal administration.

[0110] Aerosol formulations suitable for inhalation may include solids in solution and powder form, which may be combined with pharmaceutically acceptable carriers, such as inert compressed gases, for example, nitrogen.

[0111] This also includes solid formulations intended to be converted into a liquid form for oral or parenteral administration shortly before use. Such liquid forms include solutions, suspensions, and emulsions.

[0112] Another embodiment provides the use of compositions of compounds of formula I-II or pharmaceutically acceptable salts thereof formulated for transdermal delivery. The transdermal compositions may be in the form of creams, lotions, aerosols, and / or emulsions, and may be contained in matrix or reservoir-type transdermal patches, which are conventionally used in the art for this purpose.

[0113] Another embodiment provides the use of formulating compositions comprising compounds of formula I-II or pharmaceutically acceptable salts thereof for subcutaneous delivery. Another embodiment provides the use of compositions suitable for oral delivery. In some embodiments, it may be advantageous to prepare pharmaceutical formulations comprising one or more compounds of formula I-II or pharmaceutically acceptable salts thereof into unit dosage forms. In such forms, the formulation is subdivided into appropriately sized unit doses containing a suitable amount of the active ingredient (e.g., an effective amount to achieve the desired purpose). Each of the foregoing alternatives is considered to be included in the various embodiments of this disclosure.

[0114] When used in combination with one or more other therapeutic agents (“combination therapy”), the compounds used in the methods of this disclosure, namely compounds of formulas I-II, may be administered together or sequentially. When administered sequentially, the compounds of this disclosure may be administered before or after one or more other therapeutic agents, as determined by those skilled in the art or according to patient preference.

[0115] If formulated into a fixed dose, such a combination product uses a compound of formula I-II or a pharmaceutically acceptable salt thereof within the dose range described herein, and other pharmaceutically active agents or treatments within that dose range.

[0116] combination therapy Another embodiment provides a treatment method using pharmaceutically acceptable compositions comprising the compounds of this disclosure, as a net chemical or optionally also comprising additional ingredients. Such compositions are intended for use alone or in combination therapy. For the preparation of pharmaceutical compositions from the compounds of this disclosure, an inert, pharmaceutically acceptable carrier may be a solid or liquid. Solid forms include powders, tablets, dispersible granules, capsules, pouches, and suppositories. Powders and tablets may contain about 5% to about 95% of the active ingredient. Suitable solid carriers are known in the art, such as magnesium carbonate, magnesium stearate, talc, sugar, or lactose. Tablets, powders, pouches, and capsules are available as solid dosage forms suitable for oral administration. Examples of pharmaceutically acceptable carriers and methods for preparing various compositions can be found in A. Gennaro (edited). Remington's Pharmaceutical Sciences Found in 18th edition (1990), Mack Publishing Co., Easton, Pennsylvania.

[0117] Non-limiting examples of other drugs and active agents that can be used in combination therapies for the treatment of malaria include the following: Coartem® (Novartis International AG, Basel, Switzerland; artemether + fluorenyl alcohol), Eurotramesim® (Sigma-Tau Pharmaceuticals, Inc., Rome, Italy; dihydroartemisinin-piperaquine), Pyramax® (ShinPoong Pharmaceutical Co., Ltd., Seoul, South Korea; phenazine-artesunate), ASAQ Winthrop® (Sanofi SA (Gentilly, France) / DNDi (Geneva, Switzerland); artesunate + amodiaquine), ASMQ (Cipla Limited (Mumbai, India) / DNDi, artesunate + mefloquine), SPAQ-CO™ (Guilin Pharmaceutical Co., Ltd. (Shanghai), amodiaquine + sulfadoxine, pyrimethamine), Artesun® (Guilin Pharmaceutical Co., Ltd., Shanghai), and others. Artesunate, artemether, artesunate, dihydroartemisinin, phenfluorene, amodiaquine, mefloquine, piperaquine, quinine, chloroquine, atovaquinone, and chloroguanidine, as well as sulfadoxine-pyrimethamine, tafenoxuron (Glaxosmithkline), OZ439 / PQP (Sanofi), OZ439 / FQ (Sanofi), KAE609 (Novartis), KAF156 (Novartis), DSM265 (NIH / Takeda), and MK-4815 (Merck & Co., Inc., Powles et al., Antimicrobial Agents and Chemotherapy 56(5):2414-2419 (2012)). The selection of such additional active ingredients is based on the existing disease or condition requiring treatment, as determined by the attending physician or other healthcare provider.

[0118] Therefore, this disclosure also provides methods for treating Plasmodium infection or malaria using compounds of formulas I-II or pharmaceutically acceptable salts thereof to inhibit plasmepsin X, plasmepsin IX, or plasmepsin X and IX, wherein the methods further include administering one or more additional antimalarial agents to a subject in need of the substance. In some embodiments, one or more additional antimalarial agents are selected from: artemether, fluorene, dihydroartemisinin, piperaquine, phenazine, artesunate, amodiaquine, mefloquine, sulfadoxine, pyrimethamine, fluorene, quinine, chloroquine, atovaquinone, and chlorguanidine.

[0119] Example The meanings of the abbreviations in the embodiments are as follows.

[0120] ACN = MeCN = CH3CN = Acetonitrile Boc = tert-butoxycarbonyl Boc2O = ditert-butyl dicarbonate Cbz = Carboxybenzyl CCl4 = Carbon tetrachloride Celite = diatomaceous earth Conc. = concentrated DBU = 1,8-diazabicyclo[5.4.0]undec-7-ene DCE = 1,2-dichloroethane DCM = dichloromethane DIAD = diisopropyl azodicarboxylate DMAP = 4-Dimethylaminopyridine DMF = N,N-dimethylformamide DMSO = dimethyl sulfoxide DPPA = Diphenylphosphoazide EDCI = EDC = 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Et2O = Diethyl ether EtOAc = Ethyl acetate EtOH = ethanol 2 nd Grubbs = [1,3-bis(2,4,6-trimethylphenyl)imidazolium-2-ylidene](chloro)(phenylmethylene)ruthenium, tricyclohexylphosphine or Grubbs® 2nd generation catalyst, Grubbs® catalyst M2a (C848) h = hours H2 = Hydrogen HCl = hydrochloric acid HPLC = High Performance Liquid Chromatography HG-II = (1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinedimethyl)dichloro(o-isopropoxyphenylmethylene)ruthenium H3PO4 = Phosphoric acid HOAc = Acetic acid IPA = Isopropanol iPrOH = Isopropanol LCMS or LC / MS = Liquid Chromatography-Mass Spectrometry LHMDS = LiHMDS = Bis(trimethylsilyl)aminolithium Mg = Magnesium min = minutes Me = methyl MeCN = Acetonitrile MeOH = CH3OH = methanol MgSO4 = Magnesium sulfate MnO2 = Manganese dioxide N2 = Nitrogen NaBH4 = Sodium borohydride NaOH = Sodium hydroxide Na2SO4 = Sodium sulfate NaBH3CN = Sodium borohydride NH3H2O ​​= ammonia water and ammonium hydroxide NH4OAC = Ammonium acetate Pd-C = Palladium on Carbon Pd(OH)₂ = Palladium hydroxide on carbon Pet.Ether = Petroleum ether PPh3 = Triphenylphosphine rt = room temperature SFC = Supercritical Fluid Chromatography SiO2 = Silicon dioxide TFA = Trifluoroacetic acid THF = Tetrahydrofuran TMS = Trimethylsilyl TMSOK = potassium trimethylsilyl alcohol TLC = Thin Layer Chromatography ZnBr2- Zinc(II) bromide 1 standard atmosphere [atm] = 101325 Pascals [Pa] = 14.6959488 psi The meanings of the abbreviations in nuclear magnetic resonance spectroscopy are as follows: s = singlet, d = doublet, dd = double doublet, dt = double triplet, ddd = double doublet, Sept = seventuplet, t = triplet, m = multiplet, br = broad, brs = broad singlet, q = quartet, J = coupling constant; and Hz = Hertz.

[0121] Several methods for preparing the compounds of this disclosure are described in the following schemes and examples. Starting materials and intermediates are commercially available from common catalog sources or prepared using known procedures or as otherwise described. Some generally applicable routes for compounds of Formula I are described in the following schemes. In some cases, the order of reaction steps in the scheme can be changed to promote the reaction or avoid unwanted reaction products.

[0122] Example The following examples are intended to be illustrative and should not be construed as further limitations. All drawings and references, patents, and published patent applications cited throughout this application are expressly incorporated herein by reference.

[0123] Example 1 Step 1: Magnesium (1.393 g, 57.3 mmol) was added to a flask under an argon atmosphere, followed by anhydrous THF (40 mL). To initiate the Grignard reaction, a few drops of 5-bromopent-1-ene (7.32 g, 49.1 mmol) were added. The reaction mixture was then refluxed (70 °C) for 1 h. After cooling to -50 °C, 2-hydroxybenzaldehyde (2 g, 16.38 mmol) from THF (10 mL) was added to the reaction mixture. The reaction was monitored by TLC. After stirring at -50 °C for 1 h, the reaction mixture was quenched with HCl (1 M, 20 mL), diluted with EtOAc (50 mL), and washed with water (50 mL). The organic layer was dried over MgSO4 and filtered through diatomaceous earth. The filtrate was concentrated, and the resulting residue was purified by rapid silica gel chromatography (ISCO®, Agela® Flash Column Silica-CS (4 g), eluent gradient of 0-10% ethyl acetate / petroleum ether, 30 mL / min). The fractions were concentrated to obtain 2-(1-hydroxyhex-5-en-1-yl)phenol.

[0124] 1 ¹H NMR (400 MHz, chloroform-d) δ 7.93 (s, ¹H), 7.16 (t, ¹H) J = 7.67 Hz, 1H), 6.93 (d, J = 7.34 Hz, 1H), 6.79-6.88 (m, 2H), 5.72-5.82 (m, 1H), 4.92-5.03 (m, 2H), 4.83 (t, J = 6.85 Hz, 1H), 2.65 (s, 1H), 2.08 (q, J= 7.09 Hz, 2H), 1.75-1.99 (m, 2H), 1.36-1.60 (m, 2H) ppm.

[0125] Step 2: Preparation of 1-(2-hydroxyphenyl)hex-5-en-1-one To a solution of 2-(1-hydroxyhexyl-5-en-1-yl)phenol (2.0 g, 10.40 mmol) in DCM (15 mL), manganese oxide (IV) (9.04 g, 104 mmol) was added, followed by MgSO4 (200 mg). The reaction mixture was then stirred at 25 °C. After 3 hours, the reaction mixture was filtered, and the filtrate was concentrated. The resulting residue was purified by rapid silica gel chromatography (ISCO®; Agela® Flash Column Silica-CS (4 g), eluent gradient of 0–10% ethyl acetate / petroleum ether, 30 mL / min). The desired fraction was concentrated to give 1-(2-hydroxyphenyl)hexyl-5-en-1-one.

[0126] MS (ESI) m / z: 191.1(M+H) + 1 H NMR (400 MHz, CDCl3) δ 8.03 (d, J = 8.0 Hz, 1H), 7.73 (t, J = 8.5 Hz, 1H), 7.25 (d, J = 8.5 Hz, 1H), 7.16 (t, J = 7.5 Hz, 1H), 6.05-6.15 (m, 1H), 5.26-5.35 (m, 2H), 3.21-3.30 (m, 2H), 2.39-2.49 (m, 2H), 2.08-2.19 (m, 2H) ppm.

[0127] Step 3: Preparation of 3-(but-3-en-1-yl)chroman-4-one A mixture of 1-(2-hydroxyphenyl)hex-5-en-1-one (550 mg, 2.89 mmol), formaldehyde (235 mg, 2.89 mmol), and 0.5 N NaOH (12 mL) was stirred at 25 °C. After 10 hours, the reaction mixture was acidified to pH 4 with 6 N HCl and stirred at room temperature for 1 hour. The reaction mixture was then diluted with EtOAc (20 mL) and washed with water (20 mL). The organic layer was dried over MgSO4 and filtered through diatomaceous earth. The filtrate was concentrated, and the resulting residue was purified by preparative TLC (SiO2, ethyl acetate / petroleum ether = 10:1) to give 3-(but-3-en-1-yl)chroman-4-one.

[0128] MS (ESI) m / z: 203.0(M+H) + 1 H NMR (400 MHz, CDCl3) δ 8.14 (d, J = 7.5Hz, 1H), 7.70 (t, J = 7.5Hz, 1H), 7.26 (t, J = 7.5Hz, 1H), 7.20 (d, J = 8.5Hz, 1H), 6.02-6.08 (m, 1H), 5.25-5.34 (m, 2H), 4.75-7.78 (m, 1H), 4.49-4.57 (m, 1H), 2.93-2.96 (m, 1H), 2.41-2.48 (m, 2H), 2.24-2.28 (m, 1H), 1.81-1.84 (m, 1H) ppm.

[0129] Step 4: Preparation of 3-(but-3-en-1-yl)chroman-4-amine NH4OAc (1143 mg, 14.83 mmol) was added to a stirred mixture of 3-(but-3-en-1-yl)chroman-4-one (250 mg, 1.236 mmol) and 4 Å molecular sieve (500 mg) in MeOH (30 mL) at 25 °C. After 30 min, NaBH3CN (311 mg, 4.94 mmol) was added. The mixture was stirred at 90 °C and then subjected to LC / MS. After 10 hours, the mixture was filtered and the filtrate was concentrated. The resulting crude residue was diluted with water (20 mL) and washed with EtOAc (3 × 25 mL). The combined organic matter was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC (TFA). The desired fraction was concentrated to give 3-(but-3-en-1-yl)chroman-4-amine.

[0130] MS (ESI) m / z: 187.0 (M+H) + 1 ¹H NMR (400 MHz, methanol-d⁴) δ 7.26–7.37 (m, 2H), 6.99 (q, J = 7.50 Hz, 1H),6.83-6.92 (m, 1H), 5.76-5.96 (m, 1H), 4.96-5.18 (m, 2H), 3.97-4.59 (m, 3H),2.06-2.44 (m, 3H), 1.40-1.68 (m, 2H)ppm.

[0131] Step 5: (4-(but-3-en-1-yl)-1-(3-((3-(but-3-en-1-yl)chroman-4-yl)carbamoyl) Preparation of tert-butyl benzyl)-4-ethyl-6-oxotetrahydropyrimidine-2(1H)-ylide)carbamate DIEA (0.189 mL, 1.080 mmol) was added to a THF (5 mL) solution of 3-((4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)methyl)benzoic acid (116 mg, 0.270 mmol) (crude), EDC (104 mg, 0.540 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (73.0 mg, 0.540 mmol), and 3-(but-3-en-1-yl)chromium-4-amine (71.4 mg, 0.351 mmol). The reaction was stirred at 25 °C and then subjected to LC / MS. After 2 hours, the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated. The residue was purified by preparative TLC (petroleum ether / EtOAc = 2:1) to give tert-butyl (4-(but-3-en-1-yl)-1-(3-((3-(but-3-en-1-yl)chroman-4-yl)carbamoyl)benzyl)-4-ethyl-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate.

[0132] MS (ESI) m / z 615.4(M+H) + Step 6: ((12E,9E)-14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1 (1,4)-pyrimidinosa-6(4,3)-chroma-3(1,3)-benzylcyclododecano-9-ene-12-ylidene)tert-butyl carbamate preparation To a solution of tert-butyl carbamate (140 mg, 0.228 mmol) in DCM (300 mL), [1,3-bis(2,4,6-trimethylphenyl)imidazolidine-2-ylidene](chloro)(phenylmethylene)ruthenium and tricyclohexylphosphine (37.1 mg, 0.046 mmol) were added to [1,3-bis(2,4,6-trimethylphenyl)imidazolidine-2-ylidene](chloro)(phenylmethylene)ruthenium. The reaction was stirred at 25 °C and then performed by LC / MS. After 20 hours, the reaction mixture was concentrated. The residue was purified by preparative TLC (petroleum ether / EtOAc = 2:1) to obtain tert-butyl carbamate ((12E,9E)-14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzylcyclododecano-9-ene-12-ylidene)carbamate.

[0133] MS (ESI) m / z 587.3 (M+H) + Step 7: tert-Butyl(E)-(14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1) Preparation of tert-butyl (1,4)-pyrimidinazole-6(4,3)-chromazole-3(1,3)-benzylcyclododecano-12-ylidene)carbamate To a solution of ((12E,9E)-14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzylcyclododecano-9-en-12-ylidene)carbamate (120 mg, 0.205 mmol) in MeOH (30 mL), palladium dihydroxy (28.7 mg, 0.020 mmol) was added. The reaction was stirred at 25 °C under a hydrogen atmosphere (15 psi) and then performed by LC / MS. Two hours later, the reaction mixture was filtered, and the filtrate was concentrated to give (E)-(14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzylcyclododecano-12-ylidene)carbamate tert-butyl ester.

[0134] MS (ESI) m / z 589.4 (M+H) + Step 8: (E)-(14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)- Preparation of tert-butyl pyrimidinium-6(4,3)-chromoside-3(1,3)-phenylhexacyclic dodecaneno-12-ylidene)carbamate (PEAK 3) Method via SFC (instrument SFC-14, method column DAICEL CHIRALPAK AD-H (250 mm × 30 mm; 5 μm): Conditions 0.1% NH3H2O ​​IPA, start B 30%, end B 30%; 100% B; flow rate (50 (mL / min) 50) Separation of (E)-(14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzohexacyclododecano-12-ylidene)carbamate tert-butyl carbamate to obtain (E)-(14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzohexacyclododecano-12-ylidene)carbamate tert-butyl carbamate (PEAK) 1) (E)-(14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzohexacyclododecano-12-ylidene) tert-butyl carbamate (PEAK) 2) (E)-(14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzohexacyclododecano-12-ylidene) tert-butyl carbamate (PEAK) 3) and (E)-(14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzylcyclododecano-12-ylidene) tert-butyl carbamate (PEAK 4).

[0135] MS (ESI) m / z: 589.4 (M+H) + Step 9: 14-Ethyl-12-imino-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidin-6 Preparation of (4,3)-chroma-3(1,3)-benzylhexacyclic dodecanoe-16,4-dione TFA (1 mL) was added to a mixture of (E)-(14-ethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzylcyclododecano-12-ylidene)carbamate (Example 1, Step 8, PEAK 3) (20 mg, 0.034 mmol) in DCM (3 mL), and the mixture was stirred at 28 °C. The reaction was monitored by LC / MS. After 1 hour, the reaction mixture was concentrated. The resulting residue was then purified by HPLC (Column Boston Green ODS 150 × 30 mm; 5 μm: conditions: water (0.1% TFA)-ACN, start B 30, end B 60, gradient time (10 min), 100% B hold time (2 min), flow rate (25 mL / min)). The desired fraction was concentrated to obtain 14-ethyl-12-imino-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,3)-chroma-3(1,3)-benzanecyclododecano-16,4-dione.

[0136] MS (ESI) m / z: 489.3(M+H) + 1 ¹H NMR (500 MHz, methanol-d⁴) δ 8.64 (d, J = 9.0 Hz, 1H), 7.88-7.79 (m, 2H),7.66 (d, J = 8.0 Hz, 1H), 7.49 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 8.0 Hz, 1H), 7.22-7.15 (m, 1H), 6.94 (dt, J = 1.0, 7.5 Hz, 1H), 6.82 (dd, J = 1.0, 8.0 Hz, 1H), 5.81 (d, J = 16.0 Hz, 1H), 5.53 (dd, J = 5.5, 9.0 Hz, 1H), 4.59 (d, J = 16.0 Hz,1H), 4.19-4.12 (m, 1H), 4.05 (dd, J = 9.0, 11.0 Hz, 1H), 3.05 (d,J = 16.5 Hz, 1H), 2.85 (d, J = 16.5 Hz, 1H), 2.33-2.23 (m, 1H), 1.75 (q, J = 7.5 Hz, 2H),1.60-1.43 (m, 4H), 1.43-1.35 (m, 1H), 1.34-1.11 (m, 7H), 0.99 (t, J = 7.5 Hz, 3H) ppm.

[0137] Example 2 Step 1: Preparation of (R)3-(1-azido-3-methoxypropyl)benzoate methyl ester Under a nitrogen atmosphere, DBU (4.03 mL, 26.8 mmol) and diphenylphosphine azide (5.08 mL, 26.8 mmol) were added to a 20 mL solution of (S)-3-(1-hydroxy-3-methoxypropyl)benzoate (2 g, 8.92 mmol) in toluene. The mixture was then stirred at 50 °C and subjected to LC / MS. After 16 hours, the reaction mixture was quenched with water (20 mL) and extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated. The residue was purified by rapid silica gel chromatography (ISCO®; 40 g SepaFlash® silica gel column, 15% EtOAc / petroleum ether gradient eluent, 40 mL / min). The fraction was concentrated to give methyl (R)-3-(1-azido-3-methoxypropyl)benzoate.

[0138] 1 ¹H NMR (400 MHz, chloroform-d) δ 7.96–8.12 (m, 2H), 7.41–7.58 (m, 2H), 4.76 (dd, J = 6.0, 8.4 Hz, 1H), 3.94 (s, 3H), 3.51 (ddd, J = 4.8, 7.6, 9.6 Hz, 1H), 3.28-3.41 (m, 4H), 1.91-2.13 (m, 2H) ppm.

[0139] Step 2: Preparation of (R)-3-(1-amino-3-methoxypropyl)benzoate methyl ester To a flask containing methyl (R)-3-(1-azido-3-methoxypropyl)benzoate (1.3 g, 5.22 mmol) in MeOH (13 mL), 10% Pd / C (0.555 g, 0.522 mmol) was added. The reaction mixture was stirred at 25 °C under a H2 atmosphere (balloon) and then subjected to LC / MS. After 10 hours, the reaction mixture was filtered (diatomaceous earth) and the filtrate was concentrated to give crude methyl (R)-3-(1-amino-3-methoxypropyl)benzoate, which was used without further purification.

[0140] MS (ESI) m / z : 224.1 (M+H + ) 1 ¹H NMR (400 MHz, chloroform-d) δ 8.01 (s, ¹H), 7.88–7.96 (m, ¹H), 7.55 (d, ¹H) J =7.6 Hz, 1H), 7.37-7.47 (m, 1H), 4.17 (t, J = 6.8 Hz, 1H), 3.92 (s, 3H), 3.41-3.46 (m, 1H), 3.28-3.37 (m, 4H), 1.88-1.99 (m, 2H) ppm.

[0141] Step 3: (R)-3-(11,11-dimethyl-9-oxo-7-thio-2,10-dioxa-6,8-diazadodecane- Preparation of methyl 5-yl)benzoate Sodium hydride (0.383 g, 9.58 mmol) was added dropwise to a THF (10 mL) solution of N,N-bisBoc-thiourea (1.324 g, 4.79 mmol) under a N2 atmosphere at 0 °C. After 20 min at this temperature, 2,2,2-trifluoroacetic anhydride (0.844 mL, 5.99 mmol) in THF (2 mL) was added dropwise. The mixture was stirred at 0 °C for another 20 min. Then, a solution of (R)-3-(1-amino-3-methoxypropyl)benzoate (1.07 g, 4.79 mmol) in THF (10 mL) was added dropwise at 0 °C. The reaction mixture was stirred at 25 °C and then subjected to LC / MS. After 16 hours, the reaction mixture was quenched with water (30 mL) and extracted with EtOAc (3 × 40 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The resulting residue was purified by rapid silica gel chromatography (ISCO®; 40 g SepaFlash® silica gel column, 30% EtOAc / petroleum ether gradient eluent, 50 mL / min). The desired fraction was concentrated to give methyl (R)-3-(11,11-dimethyl-9-oxo-7-thio-2,10-dioxa-6,8-diazadodecane-5-yl)benzoate.

[0142] MS (ESI) m / z 383.1 (M+H + ) 1 ¹H NMR (500 MHz, chloroform-d) δ 10.53 (br d, J = 7.5 Hz, 1H), 7.92-7.98 (m,2H), 7.87 (s, 1H), 7.50 (d, J = 7.5 Hz, 1H), 7.41-7.45 (m, 1H), 5.66-5.75 (m,1H), 3.90-3.93 (m, 3H), 3.36-3.40 (m, 1H), 3.27-3.34 (m, 4H), 2.09-2.25 (m,2H), 1.50 (s, 9H) ppm.

[0143] Step 4: 3-((5R,Z)-9-(but-3-en-1-yl)-7-((tert-butoxycarbonyl)imino)-9-ethyl-11-oxy Preparation of methyl 2,12-dioxa-6,8-diazatriadecan-5-yl)benzoate DIEA (4.47 mL, 25.10 mmol) was added to a MeCN (20 mL) solution of (R)-3-(11,11-dimethyl-9-oxo-7-thio-2,10-dioxa-6,8-diazadodecane-5-yl)benzoate (1.6 g, 4.18 mmol), EDCI (2.406 g, 12.55 mmol), and methyl 3-amino-3-ethylhept-6-enoate hydrochloride (1.206 g, 5.44 mmol). The reaction mixture was stirred at 25 °C under a N2 atmosphere and then subjected to LC / MS. After 16 hours, the reaction mixture was quenched with water (40 mL) and extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product methyl 3-((5R,Z)-9-(but-3-en-1-yl)-7-((tert-butoxycarbonyl)imino)-9-ethyl-11-oxo-2,12-dioxa-6,8-diazatriadecan-5-yl)benzoate, which was used without further purification.

[0144] MS (ESI) m / z 534.2 (M+H + ) Step 5: 3-((1R)-1-((E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl- Preparation of methyl 6-oxotetrahydropyrimidine-1(2H)-yl)-3-methoxypropyl)benzoate DBU (3.11 mL, 20.61 mmol) was added to a THF (22 mL) solution of methyl 3-((5R,Z)-9-(but-3-en-1-yl)-7-((tert-butoxycarbonyl)imino)-9-ethyl-11-oxo-2,12-dioxa-6,8-diazatriadecan-5-yl)benzoate (2.2 g, 4.12 mmol). The reaction mixture was stirred at 50 °C and then subjected to LC / MS. After 16 hours, the reaction mixture was concentrated. The residue was purified by rapid silica gel chromatography (ISCO®; 40 g SepaFlash® silica gel column, 15% EtOAc / petroleum ether gradient eluent, 50 mL / min). The fractions were concentrated to obtain methyl 3-((1R)-1-((E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)-3-methoxypropyl)benzoate.

[0145] MS (ESI) m / z 502.2 (M+H + ) 1 H NMR (400 MHz, chloroform-) d) δ 8.05 (s, 1H), 7.82 (d, J = 7.6 Hz, 1H), 7.51-7.64 (m, 1H), 7.26-7.30 (m, 1H), 6.30-6.34 (m, 1H), 5.60-5.71 (m, 1H), 4.83-5.02 (m, 2H), 3.82 (s, 3H), 3.33-3.46 (m, 2H), 3.22 (s, 3H), 2.67-2.80 (m,1H), 2.35-2.51 (m, 3H), 1.90-1.96 (m, 2H), 1.58-1.59 (m, 4H), 1.43 (d, J = 2.0Hz, 9H), 0.81-0.89 (m, 3H) ppm.

[0146] Step 6: Compound 3-((1R)-1-((E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4- Preparation of ethyl-6-oxotetrahydropyrimidine-1(2H)-yl)-3-methoxypropyl)benzoic acid Potassium trimethylsilanoate (384 mg, 2.99 mmol) was added to a THF (5 mL) solution of methyl 3-((1R)-1-((E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)-3-methoxypropyl)benzoate (250 mg, 0.498 mmol) under a N2 atmosphere. The reaction mixture was stirred at 25 °C and then subjected to LC / MS. After 40 min, H3PO4 (0.1 g / mL in THF) was added to adjust the pH to approximately 6-7. The reaction mixture was then quenched with water (20 mL) and extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product 3-((1R)-1-((E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)-3-methoxypropyl)benzoic acid, which was used without further purification.

[0147] MS (ESI) m / z 488.2(M+H + ) Step 7: ((E)-4-(but-3-en-1-yl)-4-ethyl-1-((R)-1-(3-(((3S,4R)-3-hydroxy-2,2- Dimethyl-6-vinylchromo-4-yl)carbamoyl)phenyl)-3-methoxypropyl)-6-oxotetrahydropyrimidine-2(1H-) Preparation of tert-butyl carbamate (subunit) DIEA (0.630 mL, 3.61 mmol) was added to a THF (6 mL) solution of 3-((1R)-1-((E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)-3-methoxypropyl)benzoic acid (220 mg, 0.451 mmol), EDC (432 mg, 2.256 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (183 mg, 1.354 mmol), and (3S,4R)-4-amino-2,2-dimethyl-6-vinylchroman-3-ol (119 mg, 0.541 mmol). The reaction mixture was stirred at 25 °C and then subjected to LC / MS. Sixteen hours later, the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by preparative TLC (petroleum ether / EtOAc = 2:1) to give tert-butyl ((E)-4-(but-3-en-1-yl)-4-ethyl-1-((R)-1-(3-(((3S,4R)-3-hydroxy-2,2-dimethyl-6-vinylchroman-4-yl)carbamoyl)phenyl)-3-methoxypropyl)-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate.

[0148] MS (ESI) m / z 689.5 (M+H + ) 1 ¹H NMR (400 MHz, chloroform-d) δ 7.97 (s, 1H), 7.65–7.69 (m, 2H), 7.40–7.42 (m, 1H), 7.31–7.34 (m, 1H), 6.85 (d, J = 8.4 Hz, 1H), 6.61-6.68 (m, 1H), 6.53-6.57 (m, 1H), 6.40-6.43 (m, 1H), 5.69-5.80 (m, 1H), 5.61 (d, J = 17.6 Hz, 1H), 5.23 (t, J = 8.0 Hz, 1H), 5.15 (d, J = 10.8 Hz, 1H), 4.96-5.08 (m, 2H), 4.57 (brs, 1H), 3.79 (dd, J= 9.2, 2.4 Hz, 1H), 3.41-3.54 (m, 2H), 3.29 (s, 3H), 2.80(br s, 1H), 2.42-2.62 (m, 3H), 2.00-2.04 (m, 1H), 1.60-1.75 (m, 4H), 1.52 (s,3H), 1.46 (s, 9H), 1.32 (s, 3H), 0.87-0.94 (m, 3H) ppm.

[0149] Step 8: Compound ((12E,63S,64R,2R,7E)-14-ethyl-63-hydroxy-2-(2-methoxyethyl)-62, 6,2-Dimethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,6)-chromanol Preparation of tert-butyl carbamate (3-(1,3)-phenylheterocyclic decaphen-7-ene-12-ylidene) To a DCE (200 mL) solution of ((E)-4-(but-3-en-1-yl)-4-ethyl-1-((R)-1-(3-(((3S,4R)-3-hydroxy-2,2-dimethyl-6-vinylchroman-4-yl)carbamoyl)phenyl)-3-methoxypropyl)-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate (250 mg, 0.363 mmol), ruthenium(VI) chloride (22.74 mg, 0.036 mmol) was added to (200 mL) of ((E)-4-(but-3-en-yl)-4-ethyl-1-((R)-1-(3-(((3S,4R)-3-hydroxy-2,2-dimethyl-6-vinylchroman-4-yl)carbamoyl)phenyl)-3-methoxypropyl)-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate (250 mg, 0.363 mmol). The reaction mixture was stirred at 50 °C under a N2 atmosphere and then subjected to LC / MS. After 16 hours, the reaction mixture was concentrated. The resulting residue was purified by rapid silica gel chromatography (ISCO®; 4 g SepaFlash® silica gel column, 15% EtOAc / petroleum ether gradient eluent, 30 mL / min). The desired fraction was concentrated to give tert-butyl carbamate ((12E,63S,64R,2R,7E)-14-ethyl-63-hydroxy-2-(2-methoxyethyl)-62,62-dimethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,6)-chroma-3(1,3)-benzocyclodesaban-7-ene-12-ylidene)carbamate (PEAK). 1) and ((12E,63S,64R,2R,7E)-14-ethyl-63-hydroxy-2-(2-methoxyethyl)-62,62-dimethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,6)-chroma-3(1,3)-benzocyclodesaban-7-ene-12-ylidene) tert-butyl carbamate (PEAK 2).

[0150] MS (ESI) m / z : 661.2 (M+H+ ) PEAK 1: 1 ¹H NMR (400 MHz, chloroform- d ) δ 7.88 (d, J J = 7.6 Hz, 1H), 7.69 (d, J J = 8.0 Hz, 1H), 7.39 - 7.43 (m, 1H), 7.21 - 7.27 (m, 2H), 6.93 - 7.00 (m, 1H), 6.66 - 6.73 (m, 1H), 6.26 - 6.40 (m, 2H), 6.07 - 6.14 (m, 1H), 5.83 - 5.93 (m, 1H), 5.08 - 5.16 (m, 1H), 3.55 - 3.71 (m, 1H), 3.31 - 3.40 (m, 2H), 3.20 - 3.23 (m, 3H), 3.15 - 3.19 (m, 1H), 2.80 - 2.96 (m, 1H), 2.48 - 2.56 (m, 1H), 2.34 - 2.48 (m, 2H), 2.15 - 2.23 (m, 1H), 1.44 - 1.47 (m, 4H), 1.32 (s, 6H), 1.17 - 1.30 (m, 9H), 0.86 - 0.94 (m, 3H) ppm.

[0151] PEAK 2: 1 ¹H NMR (500 MHz, chloroform- d ) δ 7.93 - 8.01 (m, 2H), 7.74 (d, J J = 8.0 Hz, 1H), 7.50 (t, J J = 8.0 Hz, 1H), 7.21 (s, 1H), 6.95 - 7.05 (m, 1H), 6.71 (d, J J = 8.5 Hz, 1H), 6.54 (br d, J J = 8.5 Hz, 1H), 6.27 - 6.38 (m, 2H), 5.80 - 5.88 (m, 1H), 5.19 (t, J J = 9.0 Hz, 1H), 3.60 - 3.68 (m, 2H), 3.43 - 3.51 (m, 2H), 3.27 (s, 3H), 2.78 - 2.86 (m, 1H), 2.70 (d, J= 16.5 Hz, 1H), 2.45-2.57 (m, 2H), 2.23-2.40 (m,2H), 1.56-1.62 (m, 6H), 1.32 (s, 4H), 1.20 (s, 9H), 0.95 (t, J = 7.5 Hz, 3H)ppm.

[0152] Step 9: Compound ((63S,64R,2R,E)-14-ethyl-63-hydroxy-2-(2-methoxyethyl)-62,62-di) Methyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidaza-6(4,6)-chroma-3 Preparation of (1,3)-benzenehexacyclic decaban-12-ylidene)tert-butyl carbamate To a solution of ((12E,63S,64R,2R,7E)-14-ethyl-63-hydroxy-2-(2-methoxyethyl)-62,62-dimethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,6)-chroma-3(1,3)-benzocyclodesaban-7-ene-12-ylidene)carbamate (Example 2, Step 8, PEAK 2) (30 mg, 0.045 mmol) in MeOH (2 mL), 10% Pd / C (4.83 mg, 4.54 μmol) was added. The reaction mixture was degassed and backfilled with H2 (3x). The resulting mixture was stirred at 25 °C for 5 min under a hydrogen atmosphere (15 psi) and the reaction was then monitored by LC / MS. After 15 minutes, the reaction mixture was filtered, and the filtrate was concentrated to give tert-butyl carbamate ((63S,64R,2R,E)-14-ethyl-63-hydroxy-2-(2-methoxyethyl)-62,62-dimethyl-16,4-dioxo-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,6)-chroma-3(1,3)-benzocyclodesaban-12-ylidene)carbamate, which was used without further purification.

[0153] MS (ESI) m / z 663.4(M+H) + ) Step 10: (63S,64R,2R)-14-ethyl-63-hydroxy-12-imino-2-(2-methoxyethyl)-62,62- Dimethyl-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidaza-6(4,6)-chroma-3(1,3)-benzodiazepine Preparation of cyclodecabanone-16,4-dione Tert-butyl carbamate (30 mg, 0.045 mmol) in 4N HCl-dioxane (4 ml) was stirred at 25 °C and then subjected to LC / MS. After 4 hours, the reaction mixture was concentrated. The residue was purified by reversed-phase preparative HPLC (Column Boston Green ODS 150 × 30 mm; 5 μm): conditions: water (TFA)-ACN, start B 30, end B 50, gradient time (10 min); 100% B hold time (2 min), flow rate (25 ml / min). The fraction was concentrated to obtain (63S,64R,2R)-14-ethyl-63-hydroxy-12-imino-2-(2-methoxyethyl)-62,62-dimethyl-11,12,13,14,15,16-hexahydro-5-aza-1(1,4)-pyrimidinaza-6(4,6)-chroma-3(1,3)-benzocyclodesaban-16,4-dione.

[0154] MS (ESI) m / z 563.3(M+H + ) 1 H NMR (400 MHz, methanol-) d 4) δ 7.95 (d, J = 7.6 Hz, 1H), 7.85 (d, J = 8.0 Hz,1H), 7.64 (s, 1H), 7.57 (t, J = 7.6 Hz, 1H), 7.05 (s, 1H), 6.93 (dd, J = 8.4, 1.6Hz, 1H), 6.67 (d, J = 8.4 Hz, 1H), 5.19-5.30 (m, 2H), 4.92-4.95 (m, 1H), 3.81(d, J = 9.6 Hz, 1H), 3.63 (dd, J= 8.0, 3.2 Hz, 2H), 3.40 (s, 3H), 2.85-2.99 (m,2H), 2.72-2.81 (m, 1H), 2.39-2.48 (m, 2H), 1.70-1.80 (m, 2H), 1.55-1.68 (m,2H), 1.48 (s, 3H), 1.17-1.32 (m, 7H), 0.92 (t, J = 7.6 Hz, 3H) ppm.

[0155] Example 3 Step 1: Preparation of ethyl (1S,2S)-2-((R)-1-azido-3-methoxypropyl)cyclopropane-1-carboxylate Under a nitrogen atmosphere, at 0 °C, diphenylphosphine acyl azide (5.41 g, 22.25 mmol), and DIAD (4.33 mL, 22.25 mmol) were added to a THF (60 mL) solution of triphenylphosphine (5.84 g, 22.25 mmol), (1S,2S)-2-((S)-1-hydroxy-3-methoxypropyl)cyclopropane-1-carboxylic acid ethyl ester (3 g, 14.83 mmol) and ethyl cyclopropane-1-carboxylic acid. The reaction mixture was stirred at 15 °C and then subjected to LC / MS. After 16 hours, the reaction mixture was concentrated. The resulting residue was purified by rapid silica gel chromatography (ISCO®; 80 g SepaFlash® silica gel column, 0-8% EtOAc / petroleum ether gradient eluent, 60 mL / min). The desired fraction was concentrated to obtain ethyl (1S,2S)-2-((R)-1-azido-3-methoxypropyl)cyclopropane-1-carboxylate.

[0156] 1 ¹H NMR (400 MHz, chloroform-d) δ 4.15 (q, J = 7.2 Hz, 2H), 3.47-3.50 (m, 2H), 3.34 (s, 3H), 3.06-3.10 (m, 1H), 1.79-1.96 (m, 2H), 1.72-1.74 (m, 1H), 1.58-1.64 (m, 1H), 1.20-1.33 (m, 4H), 0.88-0.90 (m, 1H) ppm.

[0157] Step 2: Preparation of (1S,2S)-2-((R)-1-amino-3-methoxypropyl)cyclopropane-1-carboxylic acid ethyl ester To a solution of (1S,2S)-2-((R)-1-azido-3-methoxypropyl)cyclopropane-1-carboxylic acid ethyl ester (2.8 g, 12.32 mmol) in EtOH (50 mL), 10% Pd / C (1.311 g, 1.232 mmol) was added. The reaction mixture was degassed and backfilled with H2 (3x). The resulting mixture was stirred at 15 °C under an H2 atmosphere (15 psi). After 4 hours, the catalyst was filtered off, and the filtrate was concentrated to give (1S,2S)-2-((R)-1-amino-3-methoxypropyl)cyclopropane-1-carboxylic acid ethyl ester, which was used without further purification.

[0158] 1 ¹H NMR (500 MHz, methanol-d⁴) δ 4.10 (dd, J = 1.5, 7.0 Hz, 2H), 3.46-3.56 (m,2H), 2.23-2.32 (m, 1H), 1.72-1.81 (m, 2H), 1.57-1.66 (m, 1H), 1.33-1.42 (m,1H), 1.24 (t, J = 7.0 Hz, 3H), 1.10-1.15 (m, 1H), 0.87-0.89 (m, 1H).

[0159] Step 3: (1S,2S)-2-((R)-11,11-dimethyl-9-oxo-7-thio-2,10-dioxa-6,8-diazolium) Preparation of ethyl cyclopropane-1-carboxylate (hexadodecane-5-yl)cyclopropane-1-carboxylate Sodium hydride (1.033 g, 25.8 mmol) was added dropwise to a THF (40 mL) solution of N,N-bisBoc-thiourea (3.57 g, 12.92 mmol) under a N2 atmosphere. After 1 hour at 0 °C, 2,2,2-trifluoroacetic anhydride (2.323 mL, 16.15 mmol) in THF (10 mL) was added dropwise. The mixture was stirred at 0 °C for 1 hour. Then, a THF (10 mL) solution of ethyl (1S,2S)-2-((R)-1-amino-3-methoxypropyl)cyclopropane-1-carboxylate (2.6 g, 12.92 mmol) was added dropwise at 0 °C. The mixture was stirred at 15 °C and the reaction was monitored by LC / MS. After 12 hours, the reaction mixture was quenched with saturated aqueous NH4Cl solution (20 mL) and water (30 mL). It was then extracted with EtOAc (3 × 60 mL). The combined organic layers were washed with brine (80 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated. The resulting residue was then purified by rapid silica gel chromatography (ISCO®; 80 g SepaFlash® silica gel column, 10% EtOAc gradient eluent, 50 mL / min). The desired fraction was concentrated to give ethyl (1S,2S)-2-((R)-11,11-dimethyl-9-oxo-7-thio-2,10-dioxa-6,8-diazadodecane-5-yl)cyclopropane-1-carboxylate.

[0160] MS (ESI) m / z 361.2(M+H + ) 1 ¹H NMR (400 MHz, chloroform-d) δ 9.96 (br d, J = 8.4 Hz, 1H), 7.80 (s, 1H), 4.32-4.44 (m, 1H), 4.07-4.20 (m, 2H), 3.49-3.65 (m, 2H), 3.36 (s, 3H), 1.98-2.06 (m, 1H), 1.90-1.98 (m, 2H), 1.60-1.67 (m, 1H), 1.49 (s, 9H), 1.19-1.30 (m, 4H), 0.87-0.96 (m, 1H).

[0161] Step 4: (1S,2S)-2-((5R,9R,Z)-9-(but-3-en-1-yl)-7-((tert-butoxycarbonyl)imino)-9- Preparation of ethyl 11-oxo-2,12-dioxa-6,8-diazatriadecan-5-yl)cyclopropane-1-carboxylic acid ester Ethyl (1S,2S)-2-((R)-11,11-dimethyl-9-oxo-7-thio-2,10-dioxa-6,8-diazadodecane-5-yl)cyclopropane-1-carboxylate (700 mg, 1.942 mmol) was added to a MeCN (25 mL) solution of (R)-3-amino-3-ethylhepta-6-enoate hydrochloride (517 mg, 2.330 mmol), 3-(((ethylimino)methylene)amino)-N,N-dimethylpropyl-1-amine hydrochloride (745 mg, 3.88 mmol), and N-ethyl-N-isopropylpropane-2-amine (1.729 mL, 9.71 mmol). The mixture was stirred at 20 °C under a N2 atmosphere, and the reaction was monitored by LC / MS. After 16 hours, the reaction mixture was quenched with water (30 mL). Extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated to give crude (1S,2S)-2-((5R,9R,Z)-9-(but-3-en-1-yl)-7-((tert-butoxycarbonyl)imino)-9-ethyl-11-oxo-2,12-dioxa-6,8-diazatriadecan-5-yl)cyclopropane-1-carboxylic acid ethyl ester, which was used without further purification.

[0162] MS (ESI) m / z 512.4(M+H + ) Step 5: (1S,2S)-2-((R)-1-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imine) Preparation of ethyl 1(2H)-1-(4-ethyl-6-oxotetrahydropyrimidine-1(2H)-1-yl)-3-methoxypropyl)cyclopropane-1-carboxylic acid DBU (1.444 mL, 9.58 mmol) was added to a 20 mL solution of ethyl (1S,2S)-2-((5R,9R,Z)-9-(but-3-en-1-yl)-7-((tert-butoxycarbonyl)imino)-9-ethyl-11-oxo-2,12-dioxa-6,8-diazatriadecan-5-yl)cyclopropane-1-carboxylate (980 mg, 1.915 mmol) in THF. The reaction mixture was stirred at 50 °C and then subjected to LC / MS. After 16 hours, the reaction mixture was quenched with water (20 mL) and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated. The resulting residue was purified by rapid silica gel chromatography (ISCO®; 20 g SepaFlash® silica gel column, 15% EtOAc / petroleum ether gradient eluent, 35 mL / min). The desired fraction was concentrated to give ethyl (1S,2S)-2-((R)-1-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)-3-methoxypropyl)cyclopropane-1-carboxylate.

[0163] MS (ESI) m / z 480.3(M+H + ) 1 H NMR(400 MHz, chloroform-d) δ 10.01 (br s, 1H), 5.75-5.82 (m, 1H), 4.96-5.10 (m, 2H), 4.36-4.48 (m, 1H), 3.99-4.17 (m, 3H), 3.32-3.48 (m, 2H), 3.26(s, 3H), 2.49-2.66 (m, 3H), 2.18-2.43 (m, 2H), 2.06-2.13 (m, 2H), 1.55-1.74(m, 5H), 1.47 (br d, J = 9.6 Hz, 9H), 1.29-1.30 (m, 1H), 1.20 (t, J = 7.2 Hz, 3H), 0.96-1.03 (m, 1H), 0.89-0.95 (m, 3H) ppm.

[0164] Step 6: (1S,2S)-2-(can-1-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imine) Preparation of 1-(2H)-(3-methoxypropyl)cyclopropane-1-carboxylic acid (1-(2H)-(3-methoxypropyl)cyclopropane-1-carboxylic acid) Under a nitrogen atmosphere, potassium trimethylsilanolate (535 mg, 4.17 mmol) was added to a THF (10 mL) solution of ethyl (1S,2S)-2-((R)-1-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)-3-methoxypropyl)cyclopropane-1-carboxylate (250 mg, 0.521 mmol). The reaction mixture was stirred at 20 °C and then subjected to LC / MS. After 1.5 hours, H3PO4 (0.1 g / mL in THF) was added to adjust the pH to approximately 7-8. The reaction mixture was then quenched with water (20 mL) and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (1S,2S)-2-((R)-1-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)-3-methoxypropyl)cyclopropane-1-carboxylic acid, which was used without further purification.

[0165] MS (ESI) m / z 452.3(M+H + ) Step 7: ((R,E)-4-(but-3-en-1-yl)-1-((R)-1-((1S,2S)-2-(((S)-2,2-dimethyl-6- Vinylchromium-4-yl)carbamoyl)cyclopropyl)-3-methoxypropyl)-4-ethyl-6-oxotetrahydropyrimidine-2(1H)- Preparation of tert-butyl carbamate (subunit) DIEA (0.401 mL, 2.303 mmol) was added to a solution of (1S,2S)-2-((R)-1-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)-3-methoxypropyl)cyclopropane-1-carboxylic acid (130 mg, 0.288 mmol), EDC (276 mg, 1.439 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (78 mg, 0.576 mmol), and (S)-2,2-dimethyl-6-vinylchromium-4-amine (88 mg, 0.432 mmol) in THF (6 mL). The reaction mixture was stirred at 15 °C and then subjected to LC / MS. Sixteen hours later, the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative TLC (petroleum ether / EtOAc = 1:1) to give tert-butyl carbamate ((R,E)-4-(but-3-en-1-yl)-1-((R)-1-((1S,2S)-2-(((S)-2,2-dimethyl-6-vinylchroman-4-yl)carbamoyl)cyclopropyl)-3-methoxypropyl)-4-ethyl-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate.

[0166] MS (ESI) m / z 637.4(M+H + ) 1 ¹H NMR (400 MHz, chloroform-d) δ 9.90 (br s, 1H), 7.24–7.26 (m, 1H), 7.10 (s, 1H), 6.74 (d, J = 8.4 Hz, 1H), 6.70-6.72 (m, 1H), 5.53-5.68 (m, 2H), 5.34-5.36 (m, 2H), 5.13 (br d, J= 10.8 Hz, 1H), 4.89-4.93 (m, 2H), 4.38-4.41 (m,1H), 3.34-3.61 (m, 2H), 3.28 (s, 3H), 2.52-2.54 (m, 1H), 2.29-2.50 (m, 2H),2.08-2.23 (m, 2H), 1.89 (br s, 1H), 1.62-1.73 (m, 2H), 1.52-1.60 (m, 3H),1.47 (s, 9H), 1.43 (s, 3H), 1.29 (s, 3H), 1.25-1.27 (m, 2H), 0.97 (br s, 1H),0.86 (br t, J = 7.2 Hz, 3H).

[0167] Step 8: ((1aS,2R,6R,16aS,18aS,22E)-6-ethyl-2-(2-methoxyethyl)-15,15-dimethyl 4,18-dioxo-1a,2,5,6,7,8,15,16,16a,17,18,18a-dodecano-1H,4H-6,3-(bridged iminomethyl) (-11,13-bridged vinylcyclopropane[c]pyrano[4,3-g][1,6]diazacycloheptadecane-22-ylidene]carbamic acid Preparation of tert-butyl ester To a DCE (35 mL) solution of ((R,E)-4-(but-3-en-1-yl)-1-((R)-1-((1S,2S)-2-(((S)-2,2-dimethyl-6-vinylchroman-4-yl)carbamoyl)cyclopropyl)-3-methoxypropyl)-4-ethyl-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate (180 mg, 0.283 mmol), (1,3-di-mesothimethylimidazolidine-2-ylidene)(2-isopropoxybenzyl)ruthenium(VI) chloride (35.4 mg, 0.057 mmol) was added to (25 mL) of ((R,E)-4-(but-3-en-1-yl)-((R)-1-((1S,2S)-2-(((S)-2,2-dimethyl-6-vinylchroman-4-yl)carbamoyl)cyclopropyl)-3-methoxypropyl)-4-ethyl-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate (180 mg, 0.283 mmol). The reaction mixture was stirred at 50 °C under N2 atmosphere and then subjected to LC / MS. After 16 hours, the reaction mixture was concentrated. The residue was purified by preparative TLC (petroleum ether / EtOAc / EtOH = 8:3:1) to obtain ((1aS,2R,6R,16aS,18aS,22E)-6-ethyl-2-(2-methoxyethyl)-15,15-dimethyl-4,18-dioxo-1a,2,5,6,7,8,15,16,16a,17,18,18a-dodecylhydro-1H,4H-6,3-(bridged iminomethyl)-11,13-bridged vinylcyclopropano[c]pyrano[4,3-g][1,6]diazacycloheptadecane-22-ylidene]tert-butyl carbamate.

[0168] MS (ESI) m / z 609.4(M+H + ) Step 9: ((1aS,2R,6R,16aS,18aS,E)-6-ethyl-2-(2-methoxyethyl)-15,15-dimethyl- 4,18-dioxo-1a,2,5,6,7,8,9,10,15,16,16a,17,18,18a-tetradecano-1H,4H-6,3-(bridged imino) methyl)-11,13-bridged vinylcyclopropane[c]pyrano[4,3-g][1,6]diazacycloheptadecane-22-ylidene]aminomethyl Preparation of tert-butyl ester Add 10% Pd / C (17.48 mg, 0.016 mmol) to a solution of ((1aS,2R,6R,16aS,18aS,22E)-6-ethyl-2-(2-methoxyethyl)-15,15-dimethyl-4,18-dioxo-1a,2,5,6,7,8,15,16,16a,17,18,18a-dodecylhydro-1H,4H-6,3-(bridged iminomethyl)-11,13-bridged vinylcyclopropano[c]pyrano[4,3-g][1,6]diazacycloheptadecane-22-ylidene]carbamate tert-butyl ester (50 mg, 0.082 mmol) in MeOH (2 mL). Degas the reaction mixture and backfill with H2 (3x). Incubate the resulting mixture under a hydrogen atmosphere (15 The reaction was stirred at 20 °C and monitored by LC / MS. After 2 hours, the catalyst was filtered off. The filtrate was concentrated to give ((1aS,2R,6R,16aS,18aS,E)-6-ethyl-2-(2-methoxyethyl)-15,15-dimethyl-4,18-dioxo-1a,2,5,6,7,8,9,10,15,16,16a,17,18,18a-tetradecano-1H,4H-6,3-(bridged iminomethyl)-11,13-bridged vinylcyclopropano[c]pyrano[4,3-g][1,6]diazacycloheptadecane-22-ylidene]tert-butyl carbamate, which was used without further purification.

[0169] MS (ESI) m / z 611.4(M+H + ) Step 10: (1aS,2R,6R,16aS,18aS)-6-ethyl-22-imino-2-(2-methoxyethyl)-15,15- Dimethyl-1,1a,2,5,6,7,8,9,10,15,16,16a,17,18a-tetradecano-4H,18H-6,3-(bridged iminomethyl)- Preparation of 11,13-bridged vinylcyclopropane[c]pyrano[4,3-g][1,6]diazacycloheptadecane-4,18-dione A solution of ((1aS,2R,6R,16aS,18aS,E)-6-ethyl-2-(2-methoxyethyl)-15,15-dimethyl-4,18-dioxo-1a,2,5,6,7,8,9,10,15,16,16a,17,18,18a-tetradecano-1H,4H-6,3-(bridged iminomethyl)-11,13-bridged vinylcyclopropane[c]pyrano[4,3-g][1,6]diazacycloheptadecane-22-ylidene]carbamate tert-butyl ester (48 mg, 0.079 mmol) in HCl-dioxane (5 mL) was stirred at 25 °C. The reaction was monitored by LC / MS. After 10 hours, the reaction mixture was concentrated. The reaction mixture was analyzed by reversed-phase preparative HPLC (Column Boston Green ODS 150 × 30 mm; 5 μm: conditions: water (0.01%). TFcanACN, start B 32, end B 52, gradient time (10 min); 100% B hold time (2 min), flow rate (25 mL / min)) purification residue. Concentrate the desired fraction to obtain (1aS,2R,6R,16aS,18aS)-6-ethyl-22-imino-2-(2-methoxyethyl)-15,15-dimethyl-1,1a,2,5,6,7,8,9,10,15,16,16a,17,18a-tetradecano-4H,18H-6,3-(bridged iminomethyl)-11,13-bridged vinylcyclopropano[c]pyrano[4,3-g][1,6]diazacycloheptadecane-4,18-dione.

[0170] MS (ESI) m / z 511.3(M+H + ) 1 ¹H NMR (500 MHz, methanol-d⁴) δ 6.93 (dd, J = 2.0, 8.5 Hz, 1H), 6.83 (d, J =1.5 Hz, 1H), 6.65 (d, J = 8.0 Hz, 1H), 5.16-5.27 (m, 1H), 3.44-3.51 (m, 1H), 3.34-3.41 (m, 2H), 3.31 (s, 3H), 2.96 (d, J = 16.0 Hz, 1H), 2.73 (d, J= 16.0 Hz,1H), 2.46-2.67 (m, 4H), 2.10-2.12 (m, 1H), 2.04-2.07 (m, 1H), 1.70-1.86 (m,5H), 1.55-1.68 (m, 3H), 1.41 (s, 3H), 1.34-1.40 (m, 2H), 1.26 (s, 3H), 1.22-1.24 (m, 1H), 0.99 (t, J = 7.5 Hz, 3H), 0.89-0.91 (m, 1H) ppm.

[0171] Example 4 Step 1: Preparation of isopropyl 3-(4-oxotetrahydro-2H-pyran-3-yl)propionate The solution of tetrahydro-4H-pyran-4-one (6 g, 59.9 mmol) and pyrrolidine (5.11 g, 71.9 mmol) in toluene (80 mL) was refluxed at 130 °C using a Dean & Stark apparatus. After 6 hours, the solvent and excess pyrrolidine were removed under reduced pressure, and the crude enamine was dissolved in THF (60 mL). Isopropyl acrylate (8.21 g, 71.9 mmol) was then added, followed by heating to 90 °C for 4 hours. The solvent was then removed under reduced pressure, and the crude product was diluted with EtOAc. The organic phase was washed with dilute HCl and brine. The organic layer was then dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel rapid chromatography (ISCO; 80 g Agela silica gel rapid column, 15% EtOAc / petroleum ether gradient eluent, 50 mL / min). The desired fraction was concentrated to give isopropyl 3-(4-oxotetrahydro-2H-pyran-3-yl)propionate.

[0172] MS (ESI) m / z 215.4(M+H + ) 1 ¹H NMR (400 MHz, chloroform-d) δ 4.96–5.03 (m, 1H), 4.12–4.25 (m, 2H), 3.69–3.79 (m, 1H), 3.41 (dd, J = 10.0, 11.2 Hz, 1H), 2.54-2.66 (m, 2H), 2.23-2.46(m, 3H), 2.02-2.12 (m, 1H), 1.46-1.56 (m, 1H), 1.22 (d,J = 6.4 Hz, 6H) ppm.

[0173] Step 2: Preparation of isopropyl 3-(4-hydroxytetrahydro-2H-pyran-3-yl)propionate Isopropyl 3-(4-oxotetrahydro-2H-pyran-3-yl)propionate (2 g, 9.33 mmol) dissolved in MeOH (25 mL) was added to a 100 mL round-bottom flask. The solution was stirred at 0 °C under a N2 atmosphere. Then, NaBH4 (0.706 g, 18.67 mmol) was added in portions. After the addition, the reaction mixture was stirred at 30 °C for 1 hour. TLC showed that the reaction was complete. The mixture was slowly quenched with water (30 mL) at 0 °C under a N2 atmosphere, and then the solution was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (50 mL). The solution was dried over anhydrous Na2SO4, filtered, and concentrated. The resulting residue was purified by rapid silica gel chromatography (ISCO; 20 g Agela silica gel column, 30% EtOAc / petroleum ether gradient eluent, 30 mL / min). The fractions were concentrated to obtain isopropyl 3-(4-hydroxytetrahydro-2H-pyran-3-yl)propionate.

[0174] 1 H NMR (400 MHz, chloroform-d) δ 4.96-5.08 (m, 1H), 3.93-4.00 (m, 1H), 3.76-3.93 (m, 1H), 3.55-3.70 (m, 1H), 3.35-3.53 (m, 1H), 3.05 (dd, J = 10.0, 11.6Hz, 1H), 2.31-2.44 (m, 2H), 1.90-2.02 (m, 1H), 1.67-1.77 (m, 2H), 1.43-1.60(m, 2H), 1.24 (d, J = 6.4 Hz, 6H) ppm.

[0175] Step 3: 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxo Preparation of isopropyl tetrahydropyrimidine-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionate DIAD (2.337 mL, 12.02 mmol) was added dropwise to a THF (60 mL) solution of isopropyl 3-(4-hydroxytetrahydro-2H-pyran-3-yl)propionate (1.3 g, 6.01 mmol), (R,E)-(4-(but-3-en-1-yl)-4-ethyl-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate (1.598 g, 5.41 mmol), and Ph3P (3.15 g, 12.02 mmol). The reaction mixture was then stirred at 30 °C. LC / MS was then performed. After 12 hours, the reaction mixture was concentrated. The resulting residue was purified by rapid silica gel chromatography (ISCO; 80 g Agela silica gel column, 15% EtOAc / petroleum ether gradient eluent, 50 mL / min), followed by preparative HPLC (conditions: 0.1% FA, 75% ACN / water). The desired fraction was concentrated to give isopropyl 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionate.

[0176] MS (ESI) m / z 494.4 (M+H + ) Step 4: 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxo SFC chiral separation of isopropyl tetrahydropyrimidine-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionate Isopropyl 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionate (700 mg, 1.418 mmol) was separated by SFC (instrument SFC-16 method, SFC column Phenomenex-Cellulose-2 (250 mm × 30 mm; 10 μm): conditions: Neu-M EtOH, start B20, end B20, gradient time (18 min); 100% B hold for 1 min (1 min); flow rate (60 mL / min); 80 injections). The desired fractions were concentrated to obtain isopropyl 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionate (PEAK 1, Rt=2.347) and isopropyl 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxy)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionate (PEAK 2 & PEAK 3-yl)propionate. 3) and 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionate (PEAK 4, Rt=3.591).

[0177] MS (ESI) m / z 494.8 (M+H + ) Step 5: 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxo Preparation of tetrahydropyrimidine-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionic acid Sodium ethoxide (132 mg, 1.945 mmol) was added to a THF (3 mL) solution of isopropyl 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionate (160 mg, 0.324 mmol). The reaction mixture was stirred at 30 °C. LC / MS was then performed. After 3 hours, H3PO4 (0.1 g / mL in THF) was added to adjust the pH to approximately 6-7, and the reaction mixture was then diluted with water (5 mL) and extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated to give 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionic acid, which was used without further purification.

[0178] MS (ESI) m / z 452.3 (M+H + ) Step 6: ((4R,E)-4-(but-3-en-1-yl)-1-(3-(3-(((S)-2,2-dimethyl-6-vinylchromium-) 4-yl)amino)-3-oxopropyl)tetrahydro-2H-pyran-4-yl)-4-ethyl-6-oxotetrahydropyrimidine-2(1H)-ylide)amino Preparation of tert-butyl carbamate DIEA (0.290 mL, 1.661 mmol) was added to a solution of 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)tetrahydro-2H-pyran-3-yl)propionic acid (150 mg, 0.332 mmol), EDC (127 mg, 0.664 mmol), (S)-2,2-dimethyl-6-vinylchroman-4-amine (67.5 mg, 0.332 mmol), and 1H-benzo[d][1,2,3]triazol-1-ol (90 mg, 0.664 mmol) in THF (5 mL). The reaction was stirred at 30 °C. LC / MS was then performed. After 12 hours, the reaction mixture was quenched with water (5 ml) and extracted with EtOAc (3 × 7 mL). The combined organic layers were washed with brine (7 ml), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by preparative TLC (SiO2, PE:EA = 1:1) to give tert-butyl carbamate ((4R,E)-4-(but-3-en-1-yl)-1-(3-(3-(((S)-2,2-dimethyl-6-vinylchroman-4-yl)amino)-3-oxopropyl)tetrahydro-2H-pyran-4-yl)-4-ethyl-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate.

[0179] MS (ESI) m / z 637.4 (M+H + ) 1 H NMR (400 MHz, chloroform-) d ) δ 10.03 (br s, 1H), 7.24 (dd, J = 8.4, 2.0 Hz,1H), 7.17 (s, 1H), 6.73 (d, J= 8.4 Hz, 1H), 6.56-6.60 (m, 1H), 5.67-5.92 (m,2H), 5.51-5.61 (m, 1H), 5.22-5.30 (m, 1H), 5.02-5.13 (m, 2H), 4.97-4.99 (m,1H), 4.90 (br s, 1H), 3.94-4.04 (m, 2H), 3.44-3.46 (m, 1H), 3.14-3.16 (m,1H), 2.57-2.66 (m, 1H), 2.49-2.56 (m, 1H), 2.28-2.38 (m, 1H), 2.16-2.21 (m, 1H), 2.05-2.16 (m, 3H), 1.77-1.87 (m, 2H), 1.68-1.77 (m, 2H), 1.49-1.68 (m,5H), 1.42-1.43 (m, 13H), 1.32 (s, 3H), 0.91 (t, J = 7.6 Hz, 2H) ppm.

[0180] Step 7: ((8R,11E,18aS,26E)-8-ethyl-17,17-dimethyl-6,20-dioxo-4,4a.7,8,9, 10,17,18,18a,19,20,21,22,22a-tetradecano-1H,3H,6H-8,5-(bridged iminomethyl)-13,15-bridged ethylene [1,7]diazono[4,3-b:4',3'-h][1,7]diazacyclooctadecane-26-ylidene)carbamic acid Uncle Ding Preparation of esters To a DCE (50 mL) solution of ((4R,E)-4-(but-3-en-1-yl)-1-(3-(3-(((S)-2,2-dimethyl-6-vinylchroman-4-yl)amino)-3-oxopropyl)tetrahydro-2H-pyran-4-yl)-4-ethyl-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate (140 mg, 0.220 mmol), ruthenium(VI) chloride (13.78 mg, 0.022 mmol) was added to (2-isopropoxybenzyl)ruthenium(VI) chloride. The reaction was stirred at 50 °C with N2 continuously bubbled through the solution. After 3 hours, the reaction mixture was filtered and the filtrate was concentrated. The residue was then purified by preparative TLC (petroleum ether / EtOAc = 1:1.5) to obtain tert-butyl carbamate ((8R,11E,18aS,26E)-8-ethyl-17,17-dimethyl-6,20-dioxo-4,4a,7,8,9,10,17,18,18a,19,20,21,22,22a-tetradecano-1H,3H,6H-8,5-(bridged iminomethyl)-13,15-bridged vinyl dipyrano[4,3-b:4',3'-h][1,7]diazacyclooctadecane-26-ylidene)carbamate.

[0181] MS (ESI) m / z 609.4 (M+H + ) Step 8: ((8R,18aS,E)-8-ethyl-17,17-dimethyl-6,20-dioxo-4,4a,7,8,9,10,11, 12.17,18,18a,19,20,21,22,22a-hexadecylhydro-1H,3H,6H-8,5-(bridged iminomethyl)-13,15-ethylidene Dipyrano[4,3-b:4',3'-h][1,7]diazacyclooctadecane-26-ylidene)carbamic acid Uncle Ding Preparation of esters Pd / C (18.35 mg, 0.017 mmol) was added to a MeOH (6 mL) solution of ((8R,11E,18aS,26E)-8-ethyl-17,17-dimethyl-6,20-dioxo-4,4a,7,8,9,10,17,18,18a,19,20,21,22,22a-tetradecane-1H,3H,6H-8,5-(bridged iminomethyl)-13,15-bridged vinyl dipyrano[4,3-b:4',3'-h][1,7]diazacyclooctadecane-26-ylidene)carbamate (105 mg, 0.172 mmol). The reaction mixture was degassed and backfilled with H2 (3x). The resulting mixture was stirred at 25 °C under a H2 (15 psi) atmosphere. LC / MS was then performed. After approximately 10 minutes, the reaction mixture was filtered. The filtrate was concentrated to give tert-butyl carbamate ((8R,18aS,E)-8-ethyl-17,17-dimethyl-6,20-dioxo-4,4a,7,8,9,10,11,12,17,18,18a,19,20,21,22,22a-hexadecane-1H,3H,6H-8,5-(bridged iminomethyl)-13,15-ethylenedipyrano[4,3-b:4',3'-h][1,7]diazacyclooctadecane-26-ylidene)carbamate, which was used without further purification.

[0182] MS (ESI) m / z 611.4 (M+H + ) Step 9: (18aS)-8-ethyl-26-imino-17,17-dimethyl-3,4,4a,7,8,9,10,11,12,17, 18,18a,19,21,22,22a-hexadecylhydro-1H,6H,20H-8,5-(bridged iminomethyl)-13,15-ethylenedipyrano Preparation of [4,3-b:4',3'-h][1,7]diazacyclooctadecane-6,20-dione A dioxane (3 mL) solution of ((18aS,E)-8-ethyl-17,17-dimethyl-6,20-dioxo-4,4a,7,8,9,10,11,12,17,18,18a,19,20,21,22,22a-hexadecane-1H,3H,6H-8,5-(bridged iminomethyl)-13,15-ethylenedipyrano[4,3-b:4',3'-h][1,7]diazacyclooctadecane-26-ylidene)carbamate tert-butyl carbamate was stirred at 25 °C. The reaction was monitored by LC / MS. Four hours later, the reaction mixture was concentrated and the residue was purified by HPLC (Column Boston Prime C18 150×40 mm; 5 μm: conditions: water (0.1% TFA)-ACN, start B 30, end B50, gradient time (10 min); 100% B hold time (1 min); flow rate (25 mL / min)). The desired fraction was concentrated to give (18aS)-8-ethyl-26-imino-17,17-dimethyl-3,4,4a,7,8,9,10,11,12,17,18,18a,19,21,22,22a-hexadecane-1H,6H,20H-8,5-(bridged iminomethyl)-13,15-ethylenedipyrano[4,3-b:4',3'-h][1,7]diazacyclooctadecane-6,20-dione).

[0183] MS (ESI) m / z 511.3(M+H + ) 1 H NMR (400 MHz, methanol-) d 4) δ 6.99 (s, 1H), 6.89 (dd, J = 8.4, 1.6 Hz, 1H), 6.60 (d, J = 8.4 Hz, 1H), 5.10-5.22 (m, 1H), 4.05-4.08 (m, 1H), 3.99-4.02 (m,1H), 3.79-3.81 (m, 1H), 3.47-3.52 (m, 1H), 3.01-3.20 (m, 2H), 2.92 (d, J= 16.4Hz, 1H), 2.66-2.78 (m, 2H), 2.44-2.65 (m, 2H), 2.26-2.38 (m, 1H), 2.12-2.21(m, 1H), 2.00-2.03 (m, 1H), 1.81-1.93 (m, 2H), 1.62-1.80 (m, 7H), 1.39-1.52(m, 4H), 1.30-1.38 (m, 2H), 1.26 (s, 3H), 0.95 (t, J = 7.6 Hz, 3H) ppm.

[0184] Example 5 Step 1: Preparation of (E)-3-(4-oxo-4H-chromene-3-yl)acrylic acid A mixture of 4-oxo-4H-chromene-3-carboxaldehyde (9 g, 51.7 mmol) and malonic acid (10.76 g, 103 mmol) in pyridine (150 mL) was heated to 120 °C. TLC showed new spots. The reaction mixture was adjusted to pH 1 at 0 °C with 6N HCl aqueous solution, producing a large amount of precipitate. The mixture was then filtered, and the filter cake was dried under vacuum to give (E)-3-(4-oxo-4H-chromene-3-yl)acrylic acid, which was used without further purification.

[0185] 1 H NMR (400 MHz, DMSO-) d 6) δ 8.88 (s, 1H), 8.14 (dd, J = 8.0, 1.2 Hz, 1H),7.82-7.89 (m, 1H), 7.71 (d, J = 8.4 Hz, 1H), 7.55 (t, J = 7.6 Hz, 1H), 7.43 (d, J =16 Hz, 1H), 7.12 (d, J = 16 Hz, 1H) ppm.

[0186] Step 2: Preparation of (E)-3-(4-oxo-4H-chromene-3-yl)isopropyl acrylate H₂SO₄ (0.027 mL, 0.509 mmol) was added to a solution of (E)-3-(4-oxo-4H-chromene-3-yl)acrylic acid (11 g, 50.9 mmol) in iPrOH (150 mL), and the mixture was stirred at 90 °C for 16 hours under a nitrogen atmosphere. New spots were observed by TLC. After cooling, the mixture was diluted with a saturated aqueous solution of NaHCO₃, the pH was adjusted to 7, and the mixture was extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na₂SO₄, and filtered. The solvent was evaporated under reduced pressure to give the crude product, which was used in the next step without any further purification.

[0187] 1 H NMR (400 MHz, chloroform-) d ) δ 8.32 (dd, J = 8.0, 1.2 Hz, 1H), 8.16 (s, 1H),7.72-7.75 (m, 1H), 7.47-7.55 (m, 2H), 7.41-7.47 (m, 1H), 7.27-7.30 (m, 1H),5.13-5.21 (m, 1H), 1.34 (d, J = 6.4 Hz, 6H) ppm.

[0188] Step 3: Preparation of isopropyl 3-(4-hydroxychroman-3-yl)propionate At 25 °C and under a H2 atmosphere (50 psi), 10% Pd / C (2.472 g, 2.323 mmol) was added to a solution of (E)-3-(4-oxo-4H-chromen-3-yl)acrylate (6 g, 23.23 mmol) in iPrOH (100 mL), followed by LC / MS. After 24 hours, the catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain isopropyl 3-(4-hydroxychromen-3-yl)propionate, which was used in the next step without further purification.

[0189] MS (ESI) m / z 265.3 (M+H + ) 1 H NMR (400 MHz, chloroform-) d) δ 7.19-7.29 (m, 1H), 7.09-7.17 (m, 1H), 6.80-6.90 (m, 1H), 6.75-6.77 (m, 1H), 4.86-5.03 (m, 1H), 4.36-4.58 (m, 1H), 4.18-4.20 (m, 0.5H), 3.93-3.99 (m, 1.5H), 2.38-2.42 (m, 1H), 1.81-1.91 (m, 2H), 1.46-1.72 (m, 2H), 1.14-1.18 (m, 6H) ppm.

[0190] Step 4: 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6- Preparation of isopropyl oxotetrahydropyrimidine-1(2H)-yl)chroman-3-yl)propionate DIAD (1.125 mL, 5.67 mmol) was added to a THF (20 mL) solution of isopropyl 3-(4-hydroxychroman-3-yl)propionate (1 g, 3.78 mmol), Ph3P (1.985 g, 7.57 mmol), and (R,E)-(4-(but-3-en-1-yl)-4-ethyl-6-oxotetrahydropyrimidine-2(1H)-ylidene)carbamate (1.006 g, 3.40 mmol) under a N2 atmosphere. The reaction mixture was stirred at 20 °C and then subjected to LC / MS. After 16 hours, the reaction mixture was concentrated, and the resulting residue was purified by rapid silica gel chromatography (ISCO®; 20 g SepaFlash® silica gel column, 5-15% EtOAc / petroleum ether gradient eluent, 35 mL / min). The desired fraction was concentrated to obtain isopropyl 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)chroman-3-yl)propionate, which was then separated by SFC chiral separation.

[0191] Step 5: 3-((3S,4S)-4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4- Preparation of isopropyl ethyl-6-oxotetrahydropyrimidine-1(2H)-yl)chroman-3-yl)propionate (PEAK 2) Chiral separation: Isopropyl 3-(4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)chroman-3-yl)propionate (600 mg) was separated by passing it through an SFC1 column (DAICELCHIRALCEL OD-H (250 mm x 30 mm, 5 μm): conditions: 0.1% NH3H2O ​​EtOH start B 20%, end B 20%; gradient time 10 min; 100% B hold time 10 min; flow rate 70 mL / min). (180 injections). The desired fractions were concentrated to obtain isopropyl 3-((3R,4R)-4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)chroman-3-yl)propionate (PEAK 1) and isopropyl 3-((3S,4S)-4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)chroman-3-yl)propionate (PEAK 2).

[0192] MS (ESI) m / z : 542.3 (M+H + ) 1 H NMR (400 MHz, chloroform-) d ) δ 7.07 (t, J = 7.2 Hz, 1H), 6.85-6.91 (m, 1H), 6.76-6.84 (m, 2H), 6.18 (br d, J = 9.6 Hz, 1H), 5.81-5.83 (m, 1H), 5.02-5.12(m, 2H), 4.95-5.02 (m, 1H), 4.33-4.37 (m, 1H), 3.86 (t, J = 10.8 Hz, 1H), 2.83-2.99 (m, 1H), 2.40-2.70 (m, 4H), 2.29-2.31 (m, 1H), 2.08-2.17 (m, 2H), 1.81-1.92 (m, 1H), 1.67-1.72 (m, 4H), 1.50 (s, 9H), 1.22 (d, J = 6.4 Hz, 6H), 0.98(t, J = 7.6 Hz, 3H) ppm.

[0193] 1 H NMR (400 MHz, chloroform-) d ) δ 7.04-7.12 (m, 1H), 6.75-6.91 (m, 3H), 6.18(br d, J = 10.0 Hz, 1H), 5.80-5.82 (m, 1H), 4.92-5.14 (m, 3H), 4.35 (dd, J =11.2, 3.6 Hz, 1H), 3.86 (t, J = 10.8 Hz, 1H), 2.90-2.93 (m, 1H), 2.51-2.68 (m,2H), 2.39-2.48 (m, 1H), 2.25-2.35 (m, 1H), 2.07-2.17 (m, 2H), 1.68-1.90 (m,6H), 1.50 (s, 9H), 1.22 (d, J = 6.4 Hz, 6H), 0.99 (t, J = 7.6 Hz, 3H) ppm.

[0194] Step 6: 3-((3S,4S)-4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4- Preparation of ethyl-6-oxotetrahydropyrimidine-1(2H)-yl)chroman-3-yl)propionic acid To a solution of isopropyl 3-((3S,4S)-4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)chroman-3-yl)propionate (PEAK 2) (190 mg, 0.351 mmol) in THF (4 mL), potassium trimethylsilanolate (270 mg, 2.105 mmol) was added. The reaction mixture was stirred at 20 °C and then subjected to LC / MS. After 1 hour, the reaction mixture was adjusted to pH 6 with H3PO4 / H2O (0.1 g / mL). The mixture was quenched with water (2 mL) and then extracted with EtOAc (3 × 2 mL). The combined organic layers were washed with brine (2 mL) and then dried over anhydrous Na2SO4. The mixture was then filtered and concentrated to obtain 3-((3S,4S)-4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)chroman-3-yl)propionic acid, which was used without further purification.

[0195] MS (ESI) m / z : 500.4 (M+H + ) Step 7: ((R,E)-4-(but-3-en-1-yl)-1-((3S,4S)-3-(3-(((S)-2,2-dimethyl-6-ethylene) (4-yl)amino)-3-oxopropyl)(4-yl)-4-ethyl-6-oxotetrahydropyrimidine-2(1H)-ylide)amino Preparation of tert-butyl formate .

[0196] DIEA (0.475 mL, 2.72 mmol) was added to a solution of 3-((3S,4S)-4-((R,E)-4-(but-3-en-1-yl)-2-((tert-butoxycarbonyl)imino)-4-ethyl-6-oxotetrahydropyrimidin-1(2H)-yl)-chromium-3-yl)propionic acid (170 mg, 0.340 mmol), EDC (326 mg, 1.701 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (138 mg, 1.021 mmol), and (S)-2,2-dimethyl-6-vinylchromium-4-amine (0.346 mL, 0.340 mmol) in THF (5 mL). The reaction was stirred at 20 °C and then performed by LC / MS. Sixteen hours later, the reaction mixture was quenched with water (5 mL) and then extracted with EtOAc (3 × 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The resulting residue was purified by rapid silica gel chromatography (ISCO®; 4 g SepaFlash® silica gel column, 30% EtOAc / petroleum ether gradient eluent, 60 mL / min). The desired fraction was concentrated to give tert-butyl ((R,E)-4-(but-3-en-1-yl)-1-((3S,4S)-3-(3-(((S)-2,2-dimethyl-6-vinylchroman-4-yl)amino)-3-oxopropyl)chroman-4-yl)-4-ethyl-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate.

[0197] MS (ESI) m / z : 685.4 (M+H + ) Step 8: ((4aS,8aS,14bS,15R,18R,21E,28E)-18-ethyl-3,3-dimethyl-6,16-dioxo- 4,4a,5,6,7,8,8a,14b,17,18,19,20-dodecano-3H,9H,16H-18,15-(bridged iminomethyl)-1,23-bridged Vinylchrome[4,3-b]pyrano[4,3-h][1,7]diazacyclooctadecane-28-ylidene)tert-butyl carbamate preparation To a DCE (80 mL) solution of ((R,E)-4-(but-3-en-1-yl)-1-((3S,4S)-3-(3-(((S)-2,2-dimethyl-6-vinyl-4-yl)amino)-3-oxopropyl)-4-yl)-4-ethyl-6-oxotetrahydropyrimidin-2(1H)-ylidene)carbamate (195 mg, 0.285 mmol), ruthenium (17.84 mg, 0.028 mmol) was added to (1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinyl)dichloro(o-isopropoxyphenylmethylene)ruthenium (17.84 mg, 0.028 mmol). The reaction was stirred at 50 °C for 8 hours while continuously bubbled with N2. The reaction was monitored by LC / MS. After 8 hours, the reaction mixture was concentrated. The resulting residue was purified by rapid silica gel chromatography (ISCO®; 4 g SepaFlash® silica gel column, 30% EtOAc / petroleum ether gradient eluent, 60 mL / min). The desired fraction was concentrated to give tert-butyl carbamate ((4aS,8aS,14bS,15R,18R,21E,28E)-18-ethyl-3,3-dimethyl-6,16-dioxo-4,4a,5,6,7,8,8a,14b,17,18,19,20-dodecylhydro-3H,9H,16H-18,15-(bridged iminomethyl)-1,23-bridged vinylchromene[4,3-b]pyrano[4,3-h][1,7]diazacyclooctadecane-28-ylidene)carbamate.

[0198] MS (ESI) m / z 657.4 (M+H) + ) Step 9: ((4aS,8aS,14bS,15R,18R,E)-18-ethyl-3,3-dimethyl-6,16-dioxo-4,4a, 5,6,7,8,8a,14b,17,18,19,20,21,22-Tetradecano-3H,9H,16H-18,15-(bridged iminomethyl)-1,23- Bridged vinylchrome[4,3-b]pyrano[4,3-h][1,7]-diazacyclooctadecane-28-ylidene) tert-butyl carbamate Preparation Under a nitrogen atmosphere, 10% Pd-C (11.34 mg, 10.66 μmol) was added to a MeOH (2 mL) solution of ((4aS,8aS,14bS,15R,18R,21E,28E)-18-ethyl-3,3-dimethyl-6,16-dioxo-4,4a,5,6,7,8,8a,14b,17,18,19,20-dodecylhydro-3H,9H,16H-18,15-(bridged iminomethyl)-1,23-bridged vinylchromene[4,3-b]pyrano[4,3-h][1,7]diazacyclooctadecane-28-ylidene)carbamate (70 mg, 0.107 mmol)). The mixture was degassed and backfilled with H2 (3x). The resulting mixture was stirred under a H2 (15 psi) atmosphere at 20 °C and then subjected to LC / MS. After ~30 minutes, the reaction mixture was filtered and the filtrate was concentrated to give ((4aS,8aS,14bS,15R,18R,E)-18-ethyl-3,3-dimethyl-6,16-dioxo-4,4a,5,6,7,8,8a,14b,17,18,19,20,21,22-tetradecano-3H,9H,16H-18,15-(bridged iminomethyl)-1,23-bridged vinylchromeno[4,3-b]pyrano[4,3-h][1,7]-diazacyclooctadecane-28-ylidene) tert-butyl carbamate, which was used without further purification.

[0199] MS (ESI) m / z : 659.4 (M+H + ) Step 10: (4aS,8aS,14bS,15R,18R)-18-ethyl-28-imino-3,3-dimethyl-3,4,4a,5, 7,8,8a,14b,17,18,19,20,21,22-Tetradecano-6H,9H,16H-18,15-(bridged iminomethyl)-1,23-bridged ethyl Preparation of alkenylchrome[4,3-b]pyrano[4,3-h][1,7]diazacyclooctadecane-6,16-dione A mixture of ((4aS,8aS,14bS,15R,18R,E)-18-ethyl-3,3-dimethyl-6,16-dioxo-4,4a,5,6,7,8,8a,14b,17,18,19,20,21,22-tetradecano-3H,9H,16H-18,15-(bridged iminomethyl)-1,23-bridged vinylchromene[4,3-b]pyrano[4,3-h][1,7]-diazacyclooctadecane-28-ylidene) tert-butyl carbamate (60 mg, 0.091 mmol) and zinc(II) bromide (205 mg, 0.911 mmol) in DCM (2 mL) was stirred at 20 °C. The reaction was monitored by LC / MS. After 16 hours, the reaction mixture was concentrated. The residue was then purified by reversed-phase preparative HPLC (instrument, e.g., Method Phase separation Column Welch Xtimate C18 150 x 25 mm x 5 μm: conditions: water (0.1% TFA)-ACN, start B 26, end B 56, gradient time (11 min); 100% B hold time 2 min; flow rate (25 mL / min)). The desired fraction was concentrated to obtain (4aS,8aS,14bS,15R,18R)-18-ethyl-28-imino-3,3-dimethyl-3,4,4a,5,7,8,8a,14b,17,18,19,20,21,22-tetradecano-6H,9H,16H-18,15-(bridged iminomethyl)-1,23-bridged vinylchromeno[4,3-b]pyrano[4,3-h][1,7]diazacyclooctadecane-6,16-dione.

[0200] MS (ESI) m / z 559.3 (M+H) + ) 1 ¹H NMR (400 MHz, methanol-d⁴) δ 7.10–7.17 (m, 1H), 7.01–7.09 (m, 2H), 6.79–6.93 (m, 3H), 6.60 (d, J = 8.4 Hz, 1H), 5.23 (d, J = 10.4 Hz, 1H), 5.15-5.18 (m,1H), 4.45 (dd, J = 3.6, 11.2 Hz, 1H), 3.92 (t, J = 11.2 Hz, 1H), 3.31 (td, J= 1.6,3.2 Hz, 2H), 3.17-3.28 (m, 1H), 2.67-2.90 (m, 3H), 2.49-2.51 (m, 1H), 2.33-2.44 (m, 1H), 2.20-2.32 (m, 1H), 1.91-2.15 (m, 3H), 1.69-1.83 (m, 4H), 1.54-1.68 (m, 3H), 1.42 (s, 3H), 1.28-1.40 (m, 2H), 1.27 (s, 3H), 0.95 (t, J = 7.6Hz, 3H) ppm.

[0201] The compounds in Table 1 were prepared in a manner similar to that described in Examples 1 through 5. Isomers were separated by preparative HPLC and / or preparative chiral SFC.

[0202] The asterisk in the chemical structure diagram ( ) indicates the location of the chiral center.

[0203] Table 1

[0204] Other compounds disclosed herein are selected from: (4aS,8aS,14bS,15R,18R)-18-ethyl-28-imino-3,3-dimethyl-3,4,4a,5,7,8,8a,14b,17,18,19,20,21,22-tetradecano-6H,9H,16H-18,15-(bridged iminomethyl)-1,23-bridged vinylchromene[4,3-b]pyrano[4,3-h][1,7]diazacyclooctadecane-6,16-dione (1S,13R)-13-ethyl-11-imino-23,23-dimethyl-22-oxa-2,10,12-triazapentacyclo[16.6.2.210,13.14,8.021,25]nonacyclo-4,6,8(29),18(26),19,21(25)-hexane-3,28-dione, (1S,9R,13R)-13-ethyl-11-imino-9,23,23-trimethyl-22-oxa-2,10,12-triazapentacyclo[16.6.2.210,13.14,8.021,25]nonacyclo-4,6,8(29),18(26),19,21(25)-hexane-3,28-dione, (1S,13R)-13-ethyl-11-imino-9-(methoxymethyl)-23,23-dimethyl-22-oxa-2,10,12-triazapentacyclo[16.6.2.210,13.14,8.021,25]nonacyclo-4,6,8(29),18(26),19,21(25)-hexane-3,28-dione, (1S,9R,13R)-13-ethyl-11-imino-9-methyl-spiro[22-oxa-2,10,12-triazapentacyclo[16.6.2.210,13.14,8.021,25]nonacyclo-4,6,8(29),18(26),19,21(25)-hexane-23,1'-cyclopropane]-3,28-dione, (1S,9R,13R)-9,13-diethyl-11-imino-23,23-dimethyl-22-oxa-2,10,12-triazapentacyclo[16.6.2.210,13.14,8.021,25]nonacyclo-4,6,8(29),18(26),19,21(25)-hexane-3,28-dione, (1S,9R,13R)-13-ethyl-29-fluoro-11-imino-9,23,23-trimethyl-22-oxa-2,10,12-triazapentacyclohexane-[16.6.2.210,13.14,8.021,25]nonacyclohexane-4,6,8(29),18(26),19,21(25)-hexene-3,28-dione, (1S,9R,13R)-13-ethyl-6-fluoro-11-imino-9,23,23-trimethyl-22-oxa-2,10,12-triazapentacyclo[16.6.2.210,13.14,8.021,25]nonacyclo-4,6,8(29),18(26),19,21(25)-hexane-3,28-dione, (1S,9R,13R)-13-ethyl-11-imino-9-(2-methoxyethyl)-23,23-dimethyl-22-oxa-2,10,12-triazapentacyclo[16.6.2.210,13.14,8.021,25]nonacyclo-4,6,8(29),18(26),19,21(25)-hexane-3,28-dione; and (1S,9R,13R)-6-(difluoromethoxy)-13-ethyl-11-imino-9,23,23-trimethyl-22-oxa-2,10,12-triazapentacyclo[16.6.2.210,13.14,8.021,25]nonacyclo-4,6,8(29),18(26),19,21(25)-hexene-3,28-dione, or a pharmaceutically acceptable salt thereof.

[0205] Evaluation of antiparasitic efficacy in parasite LDH growth assay (parasite assay) Parasite stock was maintained at 4% hematocrit in RPMI-Hepes medium buffered with sodium bicarbonate and supplemented with 5% heat-inactivated human serum and 0.5% albumax.

[0206] Approximately 42 hours prior to setting up the efficacy assay, parasites were synchronized with 5% sorbitol to select for ring-stage parasites. On the day of assay setup, blood smears of parasite cultures were Giemsa stained and counted. Parasitemia was adjusted to 0.7% ring-stage, and hematocrit was diluted to 2% in RPMI-Hepes medium buffered with sodium bicarbonate and supplemented with 5% heat-inactivated human serum and 0.5% albumax. 30 μL of the diluted parasites were then added to 10 μL of medium + compound in a pre-prepared Greiner TC assay plate. The parasite assay plate was placed monolayer in an aerated humidified chamber and incubated at 37°C for 72 hours. After 72 hours of growth, the assay plate was sealed with paraffin film and frozen overnight in a single-row horizontal configuration at -80°C. The next day, the assay plate was thawed at room temperature for 4 hours, and LDH assays were performed to measure parasite growth.

[0207] Measurement of EC 50 The results are shown in Table 2.

[0208] Table 2

Claims

1. A compound having structural formula I or a pharmaceutically acceptable salt thereof: I Where A is a straight chain or a branched chain, saturated or unsaturated (C3-C) 10 A alkylene group comprising at least one -CH2- group, wherein one or more additional -CH2- groups in A are optionally and independently partially substituted by a subset selected from O, S, NR, CONR, NRCO, SO2, and SO2NR, and wherein one or more hydrogen atoms along A may be independently selected from hydroxyl, halogen, and C. 1-3 Group substitution of haloalkyl groups; X is selected from: (a) Straight or branched, saturated or unsaturated (C3-C) 10 )hydro-ion group, , , , , and (b) (c) (d) (e) (f) （g） Among them, single This indicates the connection point with the nitrogen atom of the tetrahydropyrimidine ring, while the double... Indicates the connection point with Z; J is a six-membered aryl or heteroaryl group selected from phenyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl, wherein the phenyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl are unsubstituted or substituted by one to three groups independently selected from R; G is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, bicyclobutyl, bicyclopentyl, bicyclohexyl, bicycloheptyl, bicyclononyl, pyridyl, pyrimidinyl, benzylpyrimidinyl, pyrazolyl, imidazolyl, and the group may optionally be substituted by 1 to 3 R groups; R is hydrogen, halogen, C1-C6 alkylCOOH, COOH, C3-C6 cycloalkyl, C1-C6 alkyl, halo-C1-C6 alkyl, -O halo-C1-C6 alkyl, C1-C6 alkylOH, C1-C6 alkoxy, COC1-C6 alkyl, C1-C6 alkylO-C1-C6 alkyl, heteroaryl or COOC1-C6 alkyl; Z represents a bond, -(CH2). p C(O)(CH2) p -,-phenyl-,-C 1-10 Heteroaryl-, wherein the phenyl and heteroaryl groups are optionally substituted with 1 to 3 R groups; Q is selected from: , and (a) (b) (c); Among them, single Indicates the connection point with A, while the double Indicates the connection point with Z; R 1 It is hydrogen, halogen, CN, OH, C1-C6 alkoxy, C1-C6 alkyl, C1-C6 alkyl, C1-C6 alkyl COOH, COOH, oxo, COOC1-C6 alkyl, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkyl, -C1-C6 alkyl O halo C1-C6 alkyl, halo C1-C6 alkyl, C1-C6 alkyl OH, CON(R) 2 (R) 3 ), N(R 2 (R) 3 ) or C1-C6 alkyl N(R 2 (R) 3 ); R 2 It is hydrogen, C1-C6 alkylCOOH, COOH, C3-C6 cycloalkyl, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkylOH, COC1-C6 alkyl or COOC1-C6 alkyl; R 3 It is hydrogen, C1-C6 alkylCOOH, COOH, C3-C6 cycloalkyl, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkylOH, COC1-C6 alkyl or COOC1-C6 alkyl; k is an integer from 0 to 4; and p is an integer independently selected from 0 to 4.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein A is a straight-chain or branched, saturated or unsaturated (C3-C6) hydrocarbon group.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Q is: 。 4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Q is: 。 5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Q is: 。 6. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-5, wherein Z is a bond.

7. The compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein Z is -C(O)(CH2). p - 8. The compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein Z is selected from -phenyl- and -C 3-10 The heteroaryl group is optionally substituted with 1 to 3 R groups.

9. The compound according to any one of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein X is straight-chain or branched, saturated or unsaturated (C3-C4). 10 ) Hydroxyl group.

10. The compound according to any one of claims 1-8 or a pharmaceutically acceptable salt thereof, wherein X is J is selected from fused aromatic rings of phenyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl, wherein the phenyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl are unsubstituted or substituted by 1 to 3 groups selected from R.

11. The compound according to any one of claims 1-8 or a pharmaceutically acceptable salt thereof, wherein X is 。 12. The compound according to any one of claims 1-8 or a pharmaceutically acceptable salt thereof, wherein X is G is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclobutyl, bicyclopentyl, bicyclohexyl, bicycloheptyl, and bicyclononyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclobutyl, bicyclopentyl, bicyclohexyl, bicycloheptyl, and bicyclononyl are unsubstituted or substituted by 1 to 3 R groups.

13. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-5 and 9-12, represented by structural formula II: II, Where n is an integer from 0 to 4.

14. The compound of claim 13 or a pharmaceutically acceptable salt thereof, wherein Q is: , where R 1 It is selected from hydrogen, halogen, OH, C1-C6 alkoxy, C1-C6 alkyl, -C1-halogenated C1-C6 alkyl and C1-C6 alkylOH.

15. The compound of claim 14 or a pharmaceutically acceptable salt thereof, wherein Q is: , where R 1 It is selected from hydrogen, halogen, OH, C1-C6 alkoxy, C1-C6 alkyl, -C1-halogenated C1-C6 alkyl and C1-C6 alkylOH.

16. The compound of claim 14 or a pharmaceutically acceptable salt thereof, wherein Q is: , where R 1 It is selected from hydrogen, halogen, OH, C1-C6 alkoxy, C1-C6 alkyl, -C1-halogenated C1-C6 alkyl and C1-C6 alkylOH.

17. The compound of claim 14 or a pharmaceutically acceptable salt thereof, wherein X is a straight-chain or branched, saturated or unsaturated (C3-C6) hydrocarbon group.

18. The compound according to claim 14, wherein X is J is selected from phenyl, pyridyl, and pyrimidinyl, wherein the phenyl, pyridyl, and pyrimidinyl groups are unsubstituted or substituted by 1 to 3 groups selected from R.

19. The compound according to claim 14, wherein X is 。 20. The compound of claim 14 or a pharmaceutically acceptable salt thereof, wherein X is G is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, dicyclobutyl, dicyclopentyl, phenyl, pyridyl, pyrimidinyl, benzylpyrimidinyl, pyrazolyl, and imidazolyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, dicyclobutyl, dicyclopentyl, phenyl, pyridyl, pyrimidinyl, benzylpyrimidinyl, pyrazolyl, and imidazolyl groups are unsubstituted or substituted by 1 to 3 R groups.

21. The compound according to any one of claims 1-20 or a pharmaceutically acceptable salt thereof, wherein R is selected from hydrogen, CH2COOH, (CH2)2COOH, CH(CH3)COOH, CH3, CH2CH3, (CH2)2OCH3, (CH2)3OCH3, (CH2)2OCH2CH3, (CH2)3OCH2CH3, CH2F, CHF2, CF3, (CH2)2OH and (CH2)3OH.

22. A compound selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 ， Or its pharmaceutically acceptable salt.

23. A method for treating Plasmodium infection or for treating malaria, comprising administering to a subject requiring such treatment a therapeutically effective amount of any one of claims 1-22 or a pharmaceutically acceptable salt thereof.

24. A method for inhibiting plasmepsin X, comprising administering to a subject requiring such treatment a therapeutically effective amount of the compound of any one of claims 1-22 or a pharmaceutically acceptable salt thereof.

25. A method for inhibiting plasmepsin IX, comprising administering to a subject requiring such treatment a therapeutically effective amount of the compound of any one of claims 1-22 or a pharmaceutically acceptable salt thereof.

26. A method for dual inhibition of plasmepsin X and plasmepsin IX, comprising administering to a subject requiring such treatment a therapeutically effective amount of the compound of any one of claims 1-22 or a pharmaceutically acceptable salt thereof.

27. The use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-22 for the treatment of Plasmodium infection or malaria in patients in need.

28. Use of the compound of any one of claims 1-22 or a pharmaceutically acceptable salt thereof for the inhibition of plasmepsin X in patients in need.

29. The use of the compound of any one of claims 1-22 or a pharmaceutically acceptable salt thereof for the inhibition of plasmepsin IX in patients in need.

30. The use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-22 for the inhibition of plasmepsin IX and plasmepsin X in patients in need.

31. A pharmaceutical composition comprising a compound according to any one of claims 1-22 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

32. A pharmaceutical composition comprising a compound according to any one of claims 1-22 and a pharmaceutically acceptable carrier.

33. A method for treating Plasmodium infection or for treating malaria, comprising administering the compound of any one of claims 1-22 or a pharmaceutically acceptable salt thereof and an effective amount of one or more other antimalarial agents.

34. A method of treating malaria by inhibiting plasmepsin X, IX and at least one other mechanism, comprising administering a compound of any one of claims 1-22 or a pharmaceutically acceptable salt thereof and an effective amount of an additional antimalarial agent, wherein said additional antimalarial agent acts by inhibiting a mechanism other than plasmepsin IX or plasmepsin X.

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