Compounds as immunomodulators of PD-L1 interactions

Abstract

PD-L1 inhibitors of various compound formulas are disclosed generically and specifically. Methods for preparing such PD-L1 inhibitor compounds are disclosed generically and specifically. Methods of using such PD-L1 inhibitor compounds, alone or in combination with additional agents, and compositions of such PD-L1 inhibitor compounds for the treatment of cancer and other diseases are disclosed.

Description

[Technical Field]

[0001] This application relates to the field of pharmaceutically active compounds that modulate PD-L1 protein interactions, and in particular to certain compounds, compositions and methods of use. [Background technology]

[0002] Programmed death-ligand 1 (PD-L1) is a protein that acts as a kind of "brake" to keep the body's immune response under control. PD-L1 is found on some normal cells and may be found in higher-than-normal amounts on some types of cancer cells. When PD-L1 binds to another protein called PD-1 (a protein present on T cells), it prevents T cells from killing PD-L1-containing cells, such as cancer cells. Anticancer drugs called immune checkpoint inhibitors bind to PD-L1 and block its binding. This releases the immune system's "brake," allowing T cells to freely kill cancer cells. PD-L1 may also play a key role in suppressing the adaptive arm of the immune system during certain events, such as pregnancy, tissue allotransplantation, autoimmune diseases, and other disease states, such as hepatitis.

[0003] Therefore, new compounds that inhibit the PD-1 / PD-L1 protein / protein interaction are needed. Summary of the Invention

[0004] One aspect of the present application relates to a compound of formula (I): [ka] During the ceremony, each of A and B is independently selected from the group consisting of halogen, cyano, -N3, alkyl and substituted alkyl, amine, alkylamine, and alkoxy; Z1 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Z2 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Z6 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Z7 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Each R 1 is independently -H, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, or aryl, and each R 2 is independently halogen, cyano, cycloalkyl, substituted alkyl, alkenyl, alkynyl, or aryl; Z3 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z4 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z5 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z8 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z9 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z 10 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Each R 3is independently -H, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkenyl, alkynyl, or aryl; R 4 , R 5 and R 6 each is independently halogen, cyano, cycloalkyl, substituted alkyl, alkenyl, alkenyl, alkynyl, or aryl; each of L1 and L3 is independently an alkyl, substituted alkyl, or heteroatom chain containing m atoms between ring 3 and W1, and between ring 6 and W3, where m=0, 1, 2, 3, 4, 5, or 6, and when m is 0, W1 or W3 is directly bonded to the corresponding nitrogen in ring 3 or ring 6, respectively; each of W1 and W3 is independently hydrogen, a 3-membered ring or a substituted 3-membered ring, a 4-membered ring or a substituted 4-membered ring, a 4-membered heterocyclic ring or a substituted 4-membered heterocyclic ring, a 5-membered heterocyclic ring or a substituted 5-membered heterocyclic ring, a 6-membered heterocyclic ring or a substituted 6-membered heterocyclic ring, a carboxylalkyl group or a substituted carboxylalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an ester of an amino acid, an amide of an amino acid, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid; each of L2 and L4 is independently an alkyl, substituted alkyl, or heteroatom chain containing m atoms between ring 3 and W2, and between ring 6 and W4, where m=0, 1, 2, 3, 4, 5, or 6, and when m is 0, W2 or W4 is directly bonded to the corresponding nitrogen in ring 3 or ring 6, respectively; Each of W2 and W4 is independently hydrogen, a 3-membered ring or a substituted 3-membered ring, a 4-membered ring or a substituted 4-membered ring, a 4-membered heterocyclic ring or a substituted 4-membered heterocyclic ring, a 5-membered heterocyclic ring or a substituted 5-membered heterocyclic ring, a 6-membered heterocyclic ring or a substituted 6-membered heterocyclic ring, a carboxylalkyl group or a substituted carboxylalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an ester of an amino acid, an amide of an amino acid, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid.

[0005] Another aspect of the present application relates to a method for treating a disease or condition associated with the interaction between PD-L1 and PD-1 in a subject, the method comprising the step of administering to the subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, solvate, or tautomer thereof.

[0006] Another aspect of the present application relates to a process for preparing the compounds of formula (I). DETAILED DESCRIPTION OF THE INVENTION

[0007] Reference will now be made in detail to certain aspects and exemplary embodiments of the present application, examples of which are illustrated in the accompanying structures and drawings. While aspects of the present application will be described in conjunction with exemplary embodiments, including methods, materials, and examples, such description is not intended to be limiting, and the scope of the present application is intended to encompass all equivalents, alternatives, and modifications that are commonly known or incorporated herein. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present application belongs. Those skilled in the art will recognize many techniques and materials similar or equivalent to those described herein, which can be used in the practice of aspects and embodiments of the present application. The described aspects and embodiments of the present application are not limited to the methods and materials described.

[0008] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise.

[0009] Ranges may be expressed herein as from "about" one particular value, and / or to "about" another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant in relation to the other endpoint, as well as independently of the other endpoint. It is also understood that there are a number of values ​​disclosed herein, and that each value is also herein disclosed as "about" that particular value in addition to the value itself. For example, if the value "10" is disclosed, "about 10" is also disclosed. It is also understood that when a value is disclosed, "less than or equal to" "greater than or equal to" the value, and possible ranges between the values ​​are also disclosed, as appropriately understood by one of ordinary skill in the art. For example, if the value "10" is disclosed, "less than or equal to 10" and "greater than or equal to 10" are also disclosed.

[0010] At various places in the present specification, particular features of compounds are disclosed in groups or ranges, and such disclosure is specifically intended to include every individual subcombination of the members of such groups and ranges.

[0011] The compounds described herein can be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise specified. Compounds of the present application containing asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically inactive starting materials are known in the art, such as resolution of racemic mixtures or stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present application. Cis and trans geometric isomers of the compounds of the present application are described and can be isolated as a mixture of isomers or as separated isomeric forms.

[0012] Resolution of racemic mixtures of compounds can be achieved by any of a number of methods known in the art. One method involves fractional recrystallization using a chiral resolving acid, which is an optically active, salt-forming organic acid. Suitable resolving agents for fractional recrystallization include, for example, optically active acids such as D- and L-tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, and lactic acid, or various optically active camphorsulfonic acids, such as β-camphorsulfonic acid. Other resolving agents suitable for fractional crystallization include stereomerically pure forms (e.g., S- and R-forms, or diastereomerically pure forms) of α-methylbenzylamine, 2-phenylglycinol, norephedrine, ephedrine, t-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, and the like.

[0013] Resolution of racemic mixtures can also be achieved by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). Suitable elution solvent compositions can be determined by those skilled in the art.

[0014] In compounds with two or more chiral centers, unless otherwise indicated, each chiral center in the compound can independently be either (R) or (S).

[0015] The compounds of the present application also include tautomeric forms. Tautomeric forms result from the interchange of a single bond with an adjacent double bond, accompanied by the migration of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy more than one position on the heterocyclic ring system (e.g., 1H- and 3 / f-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, H- and 2H-isoindole, and 1H- and 2 / / -pyrazole). Tautomeric forms can exist in equilibrium or can be sterically locked into one form by appropriate substitution.

[0016] The compounds of the present application may also include all isotopes of atoms present in the intermediates or final compounds. Isotopes include atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the present application may be replaced or substituted with a naturally or non-naturally occurring isotope of the atom. In some embodiments, the compounds contain at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced or substituted with deuterium. In some embodiments, the compounds contain two or more deuterium atoms.

[0017] I. Definition As used herein, the term "compound" is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted. This term is also meant to refer to compounds of the present application regardless of how they are prepared, including by synthesis, by biological process (e.g., metabolic or enzymatic transformation), or a combination thereof.

[0018] All compounds and pharmaceutically acceptable salts thereof can be found together with or isolated from other substances, such as water and solvents (e.g., hydrates and solvates). When in the solid state, the compounds described herein and their salts can exist in various forms, for example, in the form of solvates, including hydrates. The compounds can be in any solid state form, such as polymorphs or solvates, and therefore, unless otherwise specified, references herein to compounds and salts thereof should be understood to encompass any solid state form of the compound.

[0019] In some embodiments, the compounds of the present application or salts thereof are substantially isolated. By "substantially isolated," it is meant that the compound is at least partially or substantially separated from the environment in which it was formed or detected. Partial separation can include, for example, a composition enriched in the compounds of the present application.

[0020] Substantial separation can include compositions that contain at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of a compound of the present application or a salt thereof.

[0021] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit-risk ratio.

[0022] As used herein, the expressions "ambient temperature" and "room temperature" are understood in the art and generally refer to a temperature such as a reaction temperature, i.e., the temperature of the room in which a reaction is carried out, for example, a temperature of about 20°C to about 30°C.

[0023] The present application also includes pharmaceutically acceptable salts of the compounds described herein. The term "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds, in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, and the like. The pharmaceutically acceptable salts of the present application include non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or free base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol), or acetonitrile (MeCN) are preferred. Lists of suitable salts are found in Pharmaceutical Sciences, 17th Ed., (Mack Publishing Company, Easton, 1985), p. 1418, Berge et al., J. Pharm. Sci., 1977, 66(1), 1-19, and Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Wiley, 2002). In some embodiments, the compounds described herein include N-oxide forms.

[0024] The terms "individual" or "patient" are used interchangeably and refer to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, and most preferably a human.

[0025] The phrase "therapeutically effective amount" refers to that amount of an active compound or agent that elicits the biological or medicinal response in a tissue, system, animal, individual or human that is being sought by a researcher, veterinarian, physician or other clinician.

[0026] As used herein, the term "treating" or "treatment" refers to one or more of: (1) inhibiting a disease, e.g., inhibiting a disease, condition, or disorder (i.e., preventing further progression of the pathology and / or symptoms) in an individual experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder; and (2) ameliorating a disease, condition, or disorder (i.e., reversing the pathology and / or symptoms), e.g., reducing the severity of the disease, in an individual experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder.

[0027] The term "solvate" refers to the compound formed by the interaction of a solvent with EPI, its metabolites, or salts. Suitable solvates are pharmaceutically acceptable solvates, including hydrates.

[0028] As used herein, the terms "substituted" or "optionally substituted" mean that one or more hydrogen atoms of the group to which the terms "substituted" or "optionally substituted" refer has been replaced with a lower alkyl, lower aryl, lower aralkyl, lower cyclic alkyl, lower heterocycloalkyl, hydroxy, lower alkoxy, lower aryloxy, perhaloalkoxy, aralkoxy, lower heteroaryl, lower heteroaryloxy, lower heteroarylalkyl, lower heteroaralkoxy, azido, amino, halo, lower alkylthio, oxo, lower acylalkyl, lower carboxyester, carboxyl, carboxamido, nitro, lower acyloxy, lower aminoalkyl, lower alkylaminoaryl, means substituted with one of the substituents independently selected from lower alkylaryl, lower alkylaminoalkyl, lower alkoxyaryl, lower arylamino, lower aralkylamino, sulfonyl, lower carboxamidoalkylaryl, lower carboxamidoaryl, lower hydroxyalkyl, lower haloalkyl, lower alkylaminoalkylcarboxy-, lower aminocarboxamidoalkyl, cyano, lower alkoxyalkyl, lower perhaloalkyl, and lower arylalkyloxyalkyl, provided that the substitution does not exceed the normal valence of the atom being considered and that the substitution results in a stable compound, i.e., a compound that is sufficiently robust to be isolated from a reaction mixture.

[0029] The term "alkyl" refers to a straight, branched, or cyclic chain hydrocarbon radical containing only single carbon-carbon bonds. Representative examples include methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl, and cyclohexyl, all of which may be optionally substituted.

[0030] The term "aryl" refers to an aromatic group having 5 to 14 ring atoms and at least one ring having a conjugated π-electron system; the term includes carbocyclic aryl, heterocyclic aryl, and biaryl groups, all of which may be optionally substituted.

[0031] Carbocyclic aryl groups are groups having 6 to 14 ring atoms, the ring atoms on the aromatic ring being carbon atoms. Carbocyclic aryl groups include monocyclic carbocyclic aryl groups and polycyclic or fused compounds, such as optionally substituted naphthyl groups.

[0032] Heterocyclic aryl or heteroaryl groups are groups having 5 to 14 ring atoms, where 1 to 4 heteroatoms are ring atoms in an aromatic ring and the remainder are carbon atoms. Suitable heteroatoms include oxygen, sulfur, nitrogen, and selenium. Suitable heteroaryl groups include furanyl, thienyl, pyridyl, pyrrolyl, N-lower alkylpyrrolyl, pyridyl-N-oxide, pyrimidyl, pyrazinyl, imidazolyl, and the like, all of which may be optionally substituted.

[0033] The term "biaryl" refers to an aryl group having 5 to 14 atoms and containing two or more aromatic rings, including both fused ring systems and aryl groups substituted with other aryl groups. Such groups may be optionally substituted. Suitable biaryl groups include naphthyl and biphenyl.

[0034] The terms "substituted aryl" and "substituted heteroaryl" refer to aryl and heteroaryl groups substituted with one to three substituents selected from the group consisting of lower alkyl, lower alkoxy, lower perhaloalkyl, halo, hydroxy, and amino.

[0035] The term "aralkyl" refers to an alkylene group substituted with an aryl group. Suitable aralkyl groups include benzyl, picolyl, and the like, and may be optionally substituted.

[0036] The term "heteroarylalkyl" refers to an alkylene group substituted with a heteroaryl group.

[0037] The term "alkylaryl" refers to an aryl group substituted with an alkyl group. "Lower alkylaryl" refers to a group in which alkyl is lower alkyl.

[0038] The term "lower" as referred to herein in connection with organic radicals or compounds refers to 6 or fewer carbon atoms, respectively. Such groups may be straight-chained, branched, or cyclic.

[0039] The term "higher" as referred to herein in connection with organic radicals or compounds refers to seven or more carbon atoms, respectively. Such groups may be straight-chained, branched, or cyclic.

[0040] The terms "cyclic alkyl" or "cycloalkyl" refer to an alkyl group that is cyclic, having from 3 to 10 carbon atoms, and in one aspect, from 3 to 6 carbon atoms. Suitable cyclic groups include norbornyl and cyclopropyl. Such groups may be optionally substituted.

[0041] The terms "heterocycle," "heterocyclealkyl," or "heterocycloalkyl" refer to a cyclic group of 3 to 10 atoms, in one embodiment 3 to 6 atoms including at least one heteroatom, and in a further embodiment 1 to 3 heteroatoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen. The heterocyclic group may be attached via a nitrogen or carbon atom in the ring. Heterocyclic alkyl groups include unsaturated cyclic groups, fused cyclic groups, and spirocyclic groups. Suitable heterocyclic groups include pyrrolidinyl, morpholino, morpholinoethyl, and pyridyl.

[0042] The terms "arylamino" (a) and "aralkylamino" (b) each refer to the group -NRR', where, respectively, (a) R is aryl and R' is hydrogen, alkyl, aralkyl, heterocycloalkyl, or aryl, and (b) R' is aralkyl and R' is hydrogen, aralkyl, aryl, alkyl, or heterocycloalkyl.

[0043] The term "acyl" refers to -C(O)-R, where R is alkyl, heterocycloalkyl, or aryl.

[0044] The term "carboxy ester" refers to -C(O)-OR, where R is alkyl, aryl, aralkyl, cyclic alkyl, or heterocycloalkyl, all optionally substituted.

[0045] The term "carboxyl" refers to -C(O)-OH.

[0046] The term "oxo" refers to =O in an alkyl or heterocycloalkyl group.

[0047] The term "amino" refers to -NRR', where R and R' are independently selected from hydrogen, alkyl, aryl, aralkyl, and heterocycloalkyl, all except H being optionally substituted, and R and R' can form a ring system.

[0048] The term "carboxylamide" refers to -C(O)NR2, where each R is independently hydrogen or alkyl.

[0049] The term "sulfonylamido" or "-sulfonylamido" refers to -S(=O)2R2, where each R is independently hydrogen or alkyl.

[0050] The term "halogen" or "halo" refers to -F, -Cl, -Br and -I.

[0051] The term "alkylaminoalkylcarboxy" refers to the group alkyl-NR-alk-C(O)-O-, where "alk" is an alkylene group and R is H or lower alkyl.

[0052] The term "sulfonyl" or "sulfonyl" refers to -SO2R, where R is H, alkyl, aryl, aralkyl, or heterocycloalkyl.

[0053] The term "sulfonate" or "sulfonate" refers to -SO2-OR, where R is -H, alkyl, aryl, aralkyl, or heterocycloalkyl.

[0054] The term "alkenyl" refers to an unsaturated group having 2 to 12 atoms and containing at least one carbon-carbon double bond, including straight-chain, branched-chain, and cyclic groups. Alkenyl groups may be optionally substituted. Suitable alkenyl groups include allyl. "1-alkenyl" refers to an alkenyl group in which the double bond is between the first and second carbon atoms. When an 1-alkenyl group is bonded to another group, such as a W substituent bonded to a cyclic phosphonate, it is bonded to the first carbon.

[0055] The term "alkynyl" refers to an unsaturated group having 2 to 12 atoms and containing at least one carbon-carbon triple bond, including straight-chain, branched-chain, and cyclic groups. Alkynyl groups may be optionally substituted. Suitable alkynyl groups include ethynyl. "1-alkynyl" refers to an alkynyl group in which the triple bond is between the first and second carbon atoms. When an 1-alkynyl group is bonded to another group, such as a W substituent bonded to a cyclic phosphonate, it is bonded to the first carbon.

[0056] The term "alkylene" refers to a divalent straight-chain, branched-chain, or cyclic saturated aliphatic group. In one aspect, an alkylene group contains 10 or fewer atoms. In another aspect, an alkylene group contains 6 or fewer atoms. In a further aspect, an alkylene group contains 4 or fewer atoms. An alkylene group can be straight-chain, branched, or cyclic.

[0057] The term "acyloxy" refers to an ester group -OC(O)R, where R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, or heterocycloalkyl.

[0058] The term "aminoalkyl" refers to the group NR2-alk- where "alk" is an alkylene group and R is selected from -H, alkyl, aryl, aralkyl, and heterocycloalkyl.

[0059] The term "alkylaminoalkyl" refers to the group alkyl-NR-alk- where each "alk" is an independently selected alkylene and R is H or lower alkyl. "Lower alkylaminoalkyl" refers to groups in which the alkyl and alkylene groups are lower alkyl and lower alkylene, respectively.

[0060] The term "arylaminoalkyl" refers to the group aryl-NR-alk-, where "alk" is an alkylene group and R is -H, alkyl, aryl, aralkyl, or heterocycloalkyl. In "lower arylaminoalkyl," the alkylene group is lower alkylene.

[0061] The term "alkylaminoaryl-" refers to the group alkyl-NR-aryl- where "aryl" is a divalent group and R is -H, alkyl, aralkyl, or heterocycloalkyl. In "lower alkylaminoaryl," the alkyl group is lower alkyl.

[0062] The term "alkoxyaryl" refers to an aryl group substituted with an alkyloxy group. In "lower alkyloxyaryl", the alkyl group is lower alkyl.

[0063] The term "aryloxyalkyl" refers to an alkyl group substituted with an aryloxy group.

[0064] The term "aralkyloxyalkyl" refers to the group aryl-alk-O-alk- where "alk" is an alkylene group. "Lower aralkyloxyalkyl" refers to groups where the alkylene group is lower alkylene.

[0065] The terms "alkoxy-" or "alkyloxy-" refer to an alkyl-O- group.

[0066] The term "alkoxyalkyl" or "alkyloxyalkyl" refers to the group alkyl-O-alk-, where "alk" is an alkylene group. In "lower alkoxyalkyl", each alkyl and alkylene is a lower alkyl and lower alkylene, respectively.

[0067] The term "alkylthio-" refers to an alkyl-S- group.

[0068] The term "alkylthioalkyl" refers to the group alkyl-5-alk-, where "alk" is an alkylene group. In "lower alkylthioalkyl", each alkyl and alkylene is a lower alkyl and lower alkylene, respectively.

[0069] The term "alkoxycarbonyloxy-" refers to alkyl-OC(O)-O-.

[0070] The term "aryloxycarbonyloxy-" refers to aryl-OC(O)-O-.

[0071] The term "alkylthiocarbonyloxy" refers to alkyl-SC(O)-O-.

[0072] The term "amide" includes NR2-C(O)-, RC(O)-NR 1 -, NR2-S(=O)2- and RS(=O)2-NR 1 - refers to an NR group adjacent to an acyl or sulfonyl group such as in -, where R and R 1 includes -H, alkyl, aryl, aralkyl, and heterocycloalkyl.

[0073] The term "carboxamide" includes NR2-C(O)- and RC(O)-NR 1 - where R and R 1includes -H, alkyl, aryl, aralkyl, and heterocycloalkyl. This term does not include urea, -NR-C(O)-NR-.

[0074] The term "sulfonamide" or "sulfonamide" refers to NR2-S(=O)2- and RS(=O)2-NR 1 - where R and R 1 includes -H, alkyl, aryl, aralkyl, and heterocycloalkyl. This term does not include sulfonylureas, -NR-S(=O)2-NR-.

[0075] The terms "carboxamidoalkylaryl" or "carboxamidoaryl" refer to aryl-alk-NR 1 -C(O) and ar-NR 1 -C(O)-alk-, where "ar" is aryl, "alk" is alkylene, and R 1 and R includes H, alkyl, aryl, aralkyl, and heterocycloalkyl.

[0076] The terms "sulfonamidoalkylaryl" or "sulfonamidoaryl" refer to aryl-alk-NR 1 -S(=O)2- and ar-NR 1 -S(=O)2-, where "ar" is aryl, "alk" is alkylene, and R 1 and R includes —H, alkyl, aryl, aralkyl, and heterocycloalkyl.

[0077] The term "hydroxyalkyl" refers to an alkyl group substituted with one --OH.

[0078] The term "haloalkyl" refers to an alkyl group substituted with a halo.

[0079] The term "cyano" refers to -CN.

[0080] The term "nitro" refers to -NO2.

[0081] The term "acylalkyl" refers to alkyl-C(O)-alk-, where "alk" is alkylene.

[0082] The term "aminocarboxamidoalkyl" refers to the group NR2-C(O)-N(R)-alk-, where R is an alkyl group or H, and "alk" is an alkylene group. "Lower aminocarboxamidoalkyl" refers to such groups where "alk" is lower alkylene.

[0083] The term "heteroarylalkyl" refers to an alkylene group substituted with a heteroaryl group.

[0084] II. Compounds that inhibit PD-L1 activity One aspect of the present application relates to compounds that inhibit PD-L1 activity. In some embodiments, the compounds have the general structure as shown in Formula (I): 1. A compound of Formula (I), or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, solvate, or tautomer thereof, [ka] During the ceremony, each of A and B is independently selected from the group consisting of halogen, cyano, -N3, alkyl and substituted alkyl, amine, alkylamine, and alkoxy; Z1 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Z2 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Z6 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Z7 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Each R 1is independently -H, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, or aryl, and each R 2 is independently halogen, cyano, cycloalkyl, substituted alkyl, alkenyl, alkynyl, or aryl; Z3 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z4 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z5 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z8 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z9 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z 10 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Each R 3 is independently -H, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkenyl, alkynyl, or aryl; R 4 , R 5 and R 6 each is independently halogen, cyano, cycloalkyl, substituted alkyl, alkenyl, alkenyl, alkynyl, or aryl; each of L1 and L3 is independently an alkyl, substituted alkyl, or heteroatom chain containing m atoms between ring 3 and W1, and between ring 6 and W3, where m=0, 1, 2, 3, 4, 5, or 6, and when m is 0, W1 or W3 is directly bonded to the corresponding nitrogen in ring 3 or ring 6, respectively; each of W1 and W3 is independently hydrogen, a 3-membered ring or a substituted 3-membered ring, a 4-membered ring or a substituted 4-membered ring, a 4-membered heterocyclic ring or a substituted 4-membered heterocyclic ring, a 5-membered heterocyclic ring or a substituted 5-membered heterocyclic ring, a 6-membered heterocyclic ring or a substituted 6-membered heterocyclic ring, a carboxylalkyl group or a substituted carboxylalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an ester of an amino acid, an amide of an amino acid, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid; each of L2 and L4 is independently an alkyl, substituted alkyl, or heteroatom chain containing m atoms between ring 3 and W2, and between ring 6 and W4, where m=0, 1, 2, 3, 4, 5, or 6, and when m is 0, W2 or W4 is directly bonded to the corresponding nitrogen in ring 3 or ring 6, respectively; Each of W2 and W4 is independently hydrogen, a 3-membered ring or a substituted 3-membered ring, a 4-membered ring or a substituted 4-membered ring, a 4-membered heterocyclic ring or a substituted 4-membered heterocyclic ring, a 5-membered heterocyclic ring or a substituted 5-membered heterocyclic ring, a 6-membered heterocyclic ring or a substituted 6-membered heterocyclic ring, a carboxylalkyl group or a substituted carboxylalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an ester of an amino acid, an amide of an amino acid, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid.

[0085] The compounds of formula (I) may be symmetrical about the axis DD (i.e., the left-hand portion of formula (I) is a mirror image of the right-hand portion of formula (I)) or asymmetrical (i.e., the left-hand portion of formula (I) is different from the right-hand portion of formula (I)).

[0086] The compounds of the present application also include pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, solvates, enantiomers and tautomers of the compounds of formula (I).

[0087] In some embodiments, each of A and B is independently —Cl or methyl, wherein: Z1 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Z2 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Z6 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Z7 is =N-, -N(R 1 )-, =C(R 2 )- or -S-, Each R 1 is independently —H, alkyl, cycloalkyl, or substituted alkyl, and each R 2 are independently —H, —F, or methyl; Z3 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z4 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z5 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z8 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Z9 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R5 R 6 )- and Z 10 is -N=, -N(R 3 )-, =C(R 4 )- or -C(R 5 R 6 )- and Each R 3 is independently —H, alkyl, cycloalkyl, or substituted alkyl; R 4 , R 5 and R 6 each is independently halogen, cyano, cycloalkyl, or substituted alkyl; each of L1 and L3 is independently an alkyl, substituted alkyl, or heteroatom chain containing m atoms between ring 3 and W1, and between ring 6 and W3, where m=0, 1, 2, 3, 4, 5, or 6, and when m is 0, W1 or W3 is directly bonded to the corresponding nitrogen in ring 3 or ring 6, respectively; each of W1 and W3 is independently hydrogen, a 3-membered ring or a substituted 3-membered ring, a 4-membered ring or a substituted 4-membered ring, a 4-membered heterocyclic ring or a substituted 4-membered heterocyclic ring, a 5-membered heterocyclic ring or a substituted 5-membered heterocyclic ring, a 6-membered heterocyclic ring or a substituted 6-membered heterocyclic ring, a carboxylalkyl group or a substituted carboxylalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an ester of an amino acid, an amide of an amino acid, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid; each of L2 and L4 is independently an alkyl, substituted alkyl, or heteroatom chain containing m atoms between ring 3 and W2, and between ring 6 and W4, where m=0, 1, 2, 3, 4, 5, or 6, and when m is 0, W2 or W4 is directly bonded to the corresponding nitrogen in ring 3 or ring 6, respectively; Each of W2 and W4 is independently hydrogen, a 3-membered ring, a 4-membered ring, a 5-membered heterocycle or a substituted 5-membered heterocycle, a 6-membered heterocycle or a substituted 6-membered heterocycle, a carboxylalkyl group or a substituted carboxylalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an ester of an amino acid, an amide of an amino acid, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid.

[0088] In some embodiments, the combination of rings 2 and 3, and / or the combination of rings 5 ​​and 6 of the compound of Formula (I) are independently selected from the group consisting of: [ka]

[0089] In some embodiments, the combination of rings 2 and 3, and / or the combination of rings 5 ​​and 6 of the compound of Formula (I) are independently selected from the group consisting of: [ka]

[0090] In some embodiments, each of L1, L2, L3, and L4 is independently C1-C3 alkyl.

[0091] In some embodiments, each of W1, W2, W3, and W4 is independently selected from the group consisting of: [ka]

[0092] In some embodiments, one or more of W1, W2, W3, and W4 have the general formula: [ka] In the formula, R 7 and R 8Each of R is independently —H, alkyl, or substituted alkyl. 7 is one of the following: [ka] R 8 are independently —H, alkyl, or substituted alkyl.

[0093] Specific embodiments of the compounds disclosed herein are listed, without limitation, in Table 1. The compounds of the present application also include pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, solvates, or tautomers of the compounds of Table 1. [Table 1] TIFF0007776616000009.tif237170TIFF0007776616000010.tif227170TIFF00077766160 00011.tif231170TIFF0007776616000012.tif223170TIFF0007776616000013.tif140170

[0094] The present application further includes isotopically substituted compounds of the present disclosure. "Isotopically substituted" compounds are compounds of the present application in which one or more atoms have been replaced or substituted by atoms having the same atomic number but a different atomic mass or mass number, e.g., an atomic mass or mass number different from that normally found in nature (i.e., naturally occurring). It is understood that a "radiolabeled" compound is a compound incorporating at least one radioactive isotope (e.g., a radionuclide).

[0095] III. Uses of the Compounds of Formula (I) Another aspect of the present application relates to the use of compounds of formula (I), which inhibit the interaction between PD-L1 and PD-1 and are therefore useful in the treatment of diseases and disorders associated with the activity of PD-1, as well as diseases and disorders associated with PD-L1.

[0096] In some embodiments, compounds of Formula (I) promote the formation of PD-L1 dimers, thus inhibiting the interaction between PD-L1 and PD-1. In certain embodiments, compounds of the present disclosure, or pharmaceutically acceptable salts or stereoisomers thereof, are useful for therapeutic administration to enhance, stimulate, and / or increase immunity in cancer, chronic infection, or sepsis, including enhancing responses to vaccination. In some embodiments, the present disclosure provides a method for inhibiting PD-1 / PD-L1 protein / protein interaction. The method comprises administering to an individual or patient a compound of any of the formulas described herein, or a compound listed in any of the claims and described herein, or a pharmaceutically acceptable salt or stereoisomer thereof. The compounds of the present disclosure can be used alone, in combination with other drugs or therapies, or as an adjuvant or neoadjuvant, for the treatment of diseases or disorders, including cancer or infectious diseases. Any of the compounds of the present disclosure (including any of their embodiments) can be used for the uses described herein.

[0097] Compounds of the present disclosure inhibit PD-1 / PD-L1 protein / protein interaction, thereby blocking the PD-1 pathway. Blocking PD-1 can enhance immune responses against cancer cells and infectious diseases in mammals, including humans. In some embodiments, the present disclosure provides in vivo treatment of an individual or patient using a compound of any of the formulas described herein, or a salt or stereoisomer thereof, to inhibit the growth of cancerous tumors. A compound of any of the formulas described herein, or a compound recited in any of the claims and described herein, or a salt or stereoisomer thereof, can be used to inhibit the growth of cancerous tumors. Alternatively, a compound of any of the formulas described herein, or a compound recited in any of the claims and described herein, or a salt or stereoisomer thereof, can be used in combination with other agents or standard cancer treatments, as described below. In one embodiment, the present disclosure provides a method for inhibiting tumor cell growth in vitro. The method comprises contacting tumor cells in vitro with a compound of any of the formulas described herein, or a compound recited in any of the claims and described herein, or a salt or stereoisomer thereof. In another embodiment, the present disclosure provides a method of inhibiting tumor cell growth in an individual or patient, comprising administering to the individual or patient in need of treatment a therapeutically effective amount of a compound of any of the formulas described herein, or a compound listed in any of the claims and described herein, or a salt or stereoisomer thereof.

[0098] In some embodiments, provided herein are methods of treating cancer. The methods include administering to a patient in need of treatment a therapeutically effective amount of a compound of any of the formulas described herein, a compound listed in any of the claims and described herein, or a salt thereof. Examples of cancers include cancers whose growth can be inhibited using compounds of the present disclosure and cancers that typically respond to immunotherapy.

[0099] In some embodiments, the present disclosure provides a method of enhancing, stimulating, and / or increasing an immune response in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any of the formulas described herein, a compound or composition listed in any of the claims and described herein, or a salt thereof.

[0100] Examples of cancers that can be treated using the compounds of the present disclosure include, but are not limited to, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, acute myeloid leukemia, chronic myeloid leukemia, and ovarian cancer. These cancers include leukemia, acute lymphocytic leukemia, chronic or acute leukemia including chronic lymphocytic leukemia, childhood solid tumors, lymphocytic lymphoma, bladder cancer, kidney or urethral cancer, renal pelvis cancer, neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumors, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers including asbestos-induced cancer, and combinations of the above cancers. The compounds of the present disclosure are also useful for treating metastatic cancers, particularly metastatic cancers that express PD-L1.

[0101] In some embodiments, cancers treatable using the compounds of the present disclosure include melanoma (e.g., metastatic malignant melanoma, cutaneous melanoma), kidney cancer (e.g., clear cell carcinoma), prostate cancer (e.g., hormone-refractory prostate cancer), breast cancer (e.g., invasive breast cancer), colon cancer, lung cancer (e.g., non-small cell lung cancer and small cell lung cancer), head and neck squamous cell carcinoma (e.g., squamous cell carcinoma of the head and neck), urothelial cancer (e.g., bladder cancer, non-muscle-invasive bladder cancer (NMIBC)), and microsatellite instability-high (MSIhlgh) cancers.

[0102] Additionally, the present disclosure includes refractory or recurrent malignancies whose growth can be inhibited using the compounds of the present disclosure.

[0103] In some embodiments, cancers treatable using the compounds of the present disclosure include, but are not limited to, solid tumors (e.g., prostate cancer, colon cancer, esophageal cancer, endometrial cancer, ovarian cancer, uterine cancer, kidney cancer, liver cancer, pancreatic cancer, stomach cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc.), hematological cancers (e.g., leukemias such as lymphoma, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), DLBCL, mantle cell lymphoma, non-Hodgkin's lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin's lymphoma, or multiple myeloma), and combinations of the above cancers.

[0104] In some embodiments, cancers treatable using compounds of the present disclosure include, but are not limited to, cholangiocarcinoma, bile duct cancer, biliary tract cancer, triple-negative breast cancer, rhabdomyosarcoma, small cell lung cancer, leiomyosarcoma, hepatocellular carcinoma, Ewing's sarcoma, brain cancer, brain tumor, astrocytoma, neuroblastoma, neurofibroma, basal cell carcinoma, chondrosarcoma, epithelioid sarcoma, eye cancer, fallopian tube cancer, gastrointestinal cancer, gastrointestinal stromal tumor, hairy cell leukemia, intestinal cancer, pancreatic islet cell carcinoma, oral cancer, mouth cancer, pharyngeal cancer, laryngeal cancer, lip cancer, mesothelioma, cervical cancer, nasal cavity cancer, ocular cancer, ocular melanoma, pelvic cancer, rectal cancer, renal cell carcinoma, salivary gland cancer, paranasal sinus cancer, spinal cancer, tongue cancer, tubular cancer, urethral cancer, and ureteral cancer.

[0105] In some embodiments, compounds of the present disclosure can be used to treat sickle cell disease and sickle cell anemia.

[0106] In some embodiments, diseases and indications treatable using compounds of the present disclosure include, but are not limited to, hematological cancers, sarcomas, lung cancers, gastrointestinal cancers, genitourinary cancers, liver cancers, bone cancers, nervous system cancers, gynecological cancers, and skin cancers.

[0107] Exemplary hematological cancers include lymphomas and leukemias such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin's lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin's lymphoma, myeloproliferative disorders (e.g., primary myelofibrosis (PMF), polycythemia vera (PV), and essential thrombocytosis (ET)), myelodysplastic syndromes (MDS), T-cell acute lymphoblastic lymphoma (T-ALL), and multiple myeloma (MM).

[0108] Exemplary sarcomas include chondrosarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyoma, rhabdomyosarcoma, fibroma, lipoma, hamartoma, and teratoma.

[0109] Exemplary lung cancers include non-small cell lung cancer (NSCLC) (e.g., squamous cell NSCLC), small cell lung cancer, bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, chondromatous hamartoma, and mesothelioma.

[0110] Exemplary gastrointestinal cancers include esophageal cancer (carcinoma, squamous cell, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (carcinoma, lymphoma, leiomyosarcoma, adenocarcinoma), pancreatic cancer (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine cancer (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), colon cancer (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma), and colorectal cancer (e.g., colorectal adenocarcinoma).

[0111] Exemplary genitourinary cancers include kidney cancer (adenocarcinoma, Wilms' tumor [nephroblastoma]), bladder and urethral cancer (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate cancer (adenocarcinoma, sarcoma), and testicular cancer (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma). In some embodiments, the cancer is a urological cancer (e.g., papillary renal carcinoma, testicular germ cell carcinoma, chromophobe renal cell carcinoma, clear cell renal carcinoma, or prostate adenocarcinoma).

[0112] Exemplary liver cancers include hepatocarcinoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

[0113] Exemplary bone cancers include, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochondroma (osteochondral exostosis), benign chondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, and giant cell tumor.

[0114] Exemplary nervous system cancers include skull cancer (osteoma, hemangioma, granuloma, xanthomas, osteitis deformans), meningeal cancer (meningioma, meningeal sarcoma, gliomatosis), brain cancer (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cancer (neurofibroma, meningioma, glioma, sarcoma), and neuroblastoma and Lhermitte-Duclos disease.

[0115] Exemplary gynecological cancers include uterine cancer (endometrial cancer), cervical cancer (cervical carcinoma, preneoplastic cervical dysplasia), ovarian cancer (ovarian carcinoma (serous cystadenocarcinoma, serous adenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecocytoma, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma), vulvar cancer (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), vaginal cancer (clear cell carcinoma, squamous cell carcinoma, sarcoma botryoides (embryonic rhabdomyosarcoma)), and fallopian tube cancer (carcinoma).

[0116] Exemplary skin cancers include melanoma, basal cell carcinoma, squamous cell carcinoma (e.g., cutaneous squamous cell carcinoma), Kaposi's sarcoma, lenticular dysplastic nevi, lipoma, hemangioma, dermatofibroma, and keloids. In some embodiments, diseases and indications treatable using the compounds of the present disclosure include, but are not limited to, sickle cell disease (e.g., sickle cell anemia), triple-negative breast cancer (TNBC), myelodysplastic syndrome, testicular cancer, bile duct cancer, esophageal cancer, and urothelial cancer.

[0117] Blockade of the PD-1 pathway by compounds of the present disclosure can also be used to treat infectious diseases, such as viral, bacterial, fungal, and parasitic infections. The present disclosure provides a method for treating an infectious disease, such as a viral infection. The method includes administering to a patient in need of treatment a therapeutically effective amount of a compound of any of the formulas described herein, a compound listed in any of the claims and described herein, or a salt thereof. Examples of viruses that cause infectious diseases treatable by the methods of the present disclosure include, but are not limited to, human immunodeficiency virus, human papillomavirus, influenza, hepatitis A, B, C, or D virus, adenovirus, poxvirus, herpes simplex virus, human cytomegalovirus, severe acute respiratory syndrome virus, Ebola virus, and measles virus. In some embodiments, viruses causing infections treatable by the methods of the present disclosure include, but are not limited to, hepatitis viruses (types A, B, or C), herpes viruses (e.g., VZV, HSV-1, HAV-6, HSV-II, CMV, and Epstein-Barr virus), adenovirus, influenza virus, flavivirus, echovirus, rhinovirus, coxsackievirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum contagiosum virus, poliovirus, rabies virus, JC virus, Mycobacterium tuberculosis, and arboviral encephalitis virus.

[0118] The present disclosure provides a method for treating a bacterial infection. The method comprises administering to a patient in need of treatment a therapeutically effective amount of a compound of any of the formulas described herein, a compound listed in any of the claims and described herein, or a salt thereof. Non-limiting examples of pathogenic bacteria causing infections treatable by the methods of the present disclosure include chlamydia, rickettsia, mycobacteria, staphylococcus, streptococcus, pneumococcus, meningococcus, gonococcus, klebsiella, proteus, serratia, pseudomonas, legionella, diphtheria, salmonella, bacillus, cholera, tetanus, botulinum, anthrax, plague, leptospira, and lyme disease bacteria.

[0119] The present disclosure provides a method for treating a fungal infection. The method comprises administering to a patient in need of treatment a therapeutically effective amount of a compound of any of the formulas described herein, a compound listed in any of the claims and described herein, or a salt thereof. Non-limiting examples of pathogenic fungi causing infections treatable by the methods of the present disclosure include Candida (e.g., Candida albicans, Krusei, Glabrata, Tropicalis), Cryptococcus neoformans, Aspergillus (e.g., Fumigatus, Niger), Mucorales (Mucor, Absidia, Rhizopus), Sporothrix schenckii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, and Histoplasma capsulatum.

[0120] The present disclosure provides a method for treating a parasitic infection. The method comprises administering to a patient in need of treatment a therapeutically effective amount of a compound of any of the formulas described herein, a compound listed in any of the claims and described herein, or a salt thereof. Non-limiting examples of pathogenic parasites causing infections treatable by the methods of the present disclosure include Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba spp., Giardia lamblia, Cryptosporidium spp., Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondii, and Nippostrongylus brasiliensis.

[0121] The present disclosure provides a method for treating a neurodegenerative disease or disorder, the method comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any of the formulas described herein, a compound listed in any of the claims and described herein, or a salt thereof. Non-limiting examples of neurodegenerative diseases or disorders include Alzheimer's disease, Parkinson's disease, Huntington's disease, prion diseases, motor neuron diseases, spinocerebellar degeneration, and spinal muscular atrophy.

[0122] It is believed that the compounds of formula (I) or any of its embodiments may have satisfactory pharmacological profiles and promising biopharmaceutical properties, such as toxicological profiles, metabolic and pharmacokinetic properties, solubility, and permeability. It will be understood that determining appropriate biopharmaceutical properties, such as determining cytotoxicity in cells or determining the inhibition of specific targets or channels to determine potential toxicity, is within the knowledge of one of ordinary skill in the art.

[0123] In some embodiments, the compounds of the present application are useful for preventing or reducing the risk of developing any of the diseases mentioned herein, for example, for preventing or reducing the risk of developing a disease, condition or disorder in individuals who may have a predisposition to the disease, condition or disorder but who have not yet experienced or exhibited the pathology or symptoms of the disease.

[0124] Combination therapy The proliferation and survival of cancer cells can be affected by dysfunction in multiple biological pathways. Therefore, to treat such diseases, it may be useful to combine inhibitors of different mechanisms, such as enzyme inhibitors, signal transduction inhibitors, chromatin dynamics inhibitors, or immune response regulators. Targeting multiple signal transduction pathways (or multiple biomolecules involved in a given signal transduction pathway) may reduce the likelihood of drug resistance within a cell population or reduce the toxicity of treatment.

[0125] The compounds of the present disclosure can be used in combination with one or more other therapies for the treatment of diseases such as cancer or infectious diseases. Examples of diseases and indications that can be treated with combination therapy include those described herein.

[0126] Examples of cancer include solid tumors and non-solid tumors, such as liquid tumors, blood cancers. Examples of infectious diseases include viral infections, bacterial infections, fungal infections, or parasitic infections. For example, the compounds of the present disclosure can be combined with inhibitors of one or more of the following kinases for the treatment of cancer: Aktl, Akt2, Akt3, BCL2, CDK, TGF-PR, PKA, PKG, PKC, CaM-kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IDH2, IGF-1R, IR-R, PDGFotR, PDGi'PR, PI3K (alpha, beta, gamma, delta, and multiple or selective), CSF1R, KIT, FLK-1I, KDR / FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c-Met, PARP, Ron, Sea, I RKA, TRKB, TRKC, TAM kinases (Axl, Mer, Tyro3), FLT3, VEGFR / FH2, Flt4, EphAl, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK, and B-Raf. In some embodiments, the compounds of the present disclosure can be combined with one or more of the following inhibitors for the treatment of cancer or infectious diseases. Non-limiting examples of inhibitors that can be combined with the compounds of the present disclosure for the treatment of cancer and infectious diseases include FGFR inhibitors (FGFR1, FGFR2, FGFR3, or FGFR4, e.g., pemigatinib (INCY54828), INCB62079), JAK inhibitors (JAK1 and / or JAK2, e.g., ruxolitinib, baricitinib, or itacitinib (INCB39110)), IDO inhibitors (e.g., epacadostat, N LG919, or BMS-986205, MK7162), LSD1 inhibitors (e.g., INCB59872 and INCB60003), TDO inhibitors, PI3K-delta inhibitors (e.g., palsaclisib (INCB50465) and INCB50797), PI3K-gamma inhibitors such as PI3K-gamma selective inhibitors, Pirn inhibitors (e.g., INCB53914), EGFR inhibitors (also known as ErB-1 or HER-1).e.g., erlotinib, gefitinib, vandetanib, orsimertinib, cetuximab, necitumumab, or panitumumab), VEGFR inhibitors or pathway blockers (e.g., bevacizumab, pazopanib, sunitinib, sorafenib, axitinib, regorafenib, ponatinib, cabozantinib, axitinib, vandetanib, ramucirumab, lenvatinib, dib-aflibercept), PARP inhibitors (e.g., olaparib, rucaparib, veliparib, talazoparib, or niraparib), CSF1R inhibitors, TAM receptor tyrosine kinases (Tyro-3, Axl, and Mer), adenosine receptor antagonists (e.g., A2a / A2b receptor antagonists), HPK1 inhibitors, chemokine receptor inhibitors (e.g., CC1 receptor antagonists), R2 or CCR5 inhibitors), SHP1 / 2 phosphatase inhibitors, histone deacetylase inhibitors (HDACs) such as HDAC8 inhibitors, angiogenesis inhibitors, interleukin receptor inhibitors, bromo- and extra-terminal family member inhibitors (e.g., bromodomain inhibitors or BET inhibitors such as INCB54329 and INCB57643), arginase inhibitors (INCB001158), PARP inhibitors (such as rucaparib or olaparib), sitravatinib, combinations of B-Raf inhibitors and MEK inhibitors (such as encorafenib and binimetinib, dabrafenib and trametinib, or cobimetinib and vemurafenib), adenosine receptor antagonists, or combinations thereof.

[0127] In some embodiments, compounds of the present disclosure can be combined with a TLR7 agonist (eg, imiquimod).

[0128] The compounds of the present disclosure can also be used in combination with other methods of treating cancer, such as chemotherapy, radiation therapy, tumor-targeted therapy, adjuvant therapy, immunotherapy, or surgery. Examples of immunotherapy include cytokine therapy (e.g., interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, monoclonal antibodies, bispecific or multispecific antibodies, antibody-drug conjugates, adoptive T cell transfer, Toll receptor agonists, STING agonists, RIG-I agonists, oncolytic virotherapy, and immunomodulatory small molecules (including thalidomide or JAK1 / 2 inhibitors, PI3K6 inhibitors, etc.). The compounds can be administered in combination with one or more anti-cancer agents, such as chemotherapeutic agents. Examples of chemotherapeutic agents include abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezomib, intravenous busulfan, oral busulfan, calcitonin, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, Eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, hitorelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, interferon alfa 2a, irinotecan, lapatinib ditosilate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, mechlorethamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycin C, mitotane,Mitoxantrone, nandrolone phenylpropionate, nelarabine, nofetumomab, oxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, sorafenib, strontium iodide ... Examples of the anti-cancer drugs include leptozocin, sunitinib, sunitinib malate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, and zoledronate.

[0129] Other anti-cancer agents include antibody therapeutics such as trastuzumab (Herceptin), antibodies against costimulatory molecules such as CTLA-4 (e.g., ipilimumab), 4-1BB (e.g., urelumab, utomilumab), antibodies against PD-1 and PD-L1, or antibodies against cytokines (e.g., IL-10, TGF-b, etc.). Examples of antibodies against PD-1 and / or PD-L1 that can be combined with compounds of the present disclosure for the treatment of cancer or infectious diseases, such as viral, bacterial, fungal, and parasitic infections, include, but are not limited to, nivolumab, pembrolizumab, atezolizumab, durvalumab, avelumab, and SHR-1210.

[0130] The compounds of the present disclosure can be used in combination with one or more immune checkpoint inhibitors for the treatment of diseases such as cancer or infectious diseases.

[0131] Exemplary immune checkpoint inhibitors include inhibitors of immune checkpoint molecules such as CBL-B, CD27, CD28, CD40, CD122, CD96, CD73, CD47, OX40, GITR, CSF1R, JAK, PI3K delta, PI3K gamma, TAM, arginase, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, TIGIT, CD112R, VISTA, PD-1, PD-L1, and PD-L2. In some embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, and VISTA. In some embodiments, the compounds provided herein can be used in combination with one or more agents selected from a KIR inhibitor, a TIGIT inhibitor, a LAIR1 inhibitor, a CD160 inhibitor, a 2B4 inhibitor, and a TGFRbeta inhibitor.

[0132] In some embodiments, the inhibitor of an immune checkpoint molecule is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

[0133] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-1, e.g., an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), pidilizumab, SHR-1210, PDR001, or AMP-224. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD-1 monoclonal antibody is pembrolizumab.

[0134] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CTLA-4, e.g., an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab or tremelimumab.

[0135] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of LAG3, e.g., an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, or INCAGN2385.

[0136] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of TIM3, e.g., an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453, or TSR-022.

[0137] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of GITR, e.g., an anti-GITR antibody. In some embodiments, the anti-GITR antibody is TRX518, MK-4166, INCAGN1876, MK-1248, AMG228, BMS-986156, GWN323, or MEDI1873.

[0138] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of OX40, e.g., an anti-OX40 antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is MEDI0562, MOXR-0916, PF-04518600, GSK3174998, or BMS-986178. In some embodiments, the OX40L fusion protein is MEDI6383.

[0139] The compounds of the present disclosure may also be used in combination with one or more anti-inflammatory agents, steroids, immunosuppressants, or therapeutic antibodies.

[0140] A compound of any of the formulas described herein, a compound recited in any of the claims and described herein, or a salt thereof, can be combined with another immunogenic substance, such as cancer cells, purified tumor antigens (including recombinant proteins, peptides, and carbohydrate molecules), cells, and cells transfected with genes encoding immune-stimulating cytokines. Non-limiting examples of tumor vaccines that can be used include peptides of melanoma antigens (such as peptides of gplOO, MAGE antigens, Trp-2, MARTI, and / or tyrosinase), or tumor cells transfected to express the cytokine GM-CSF.

[0141] A compound of any of the formulas described herein, a compound listed in any of the claims and described herein, or a salt thereof, can be used in combination with vaccination protocols for the treatment of cancer. In some embodiments, tumor cells are transduced to express GM-CSF. In some embodiments, tumor vaccines include proteins derived from viruses implicated in human cancers, such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV), and Kaposi's herpes sarcoma virus (KHSV). In some embodiments, compounds of the present disclosure can be used in combination with tumor-specific antigens, such as heat shock proteins isolated from the tumor tissue itself. In some embodiments, a compound of any of the formulas described herein, a compound listed in any of the claims and described herein, or a salt thereof can be combined with dendritic cell immunization to activate a potent anti-tumor response.

[0142] The compounds of the present disclosure can be used in combination with bispecific macrocyclic peptides that target effector cells expressing Fe alpha or Fe gamma receptors to tumor cells. The compounds of the present disclosure can also be combined with macrocyclic peptides that activate the host's immune responsiveness.

[0143] The compounds of the present disclosure can be used in combination with bone marrow transplantation for the treatment of various tumors of hematopoietic origin.

[0144] A compound of any of the formulas described herein, a compound listed in any of the claims and described herein, or a salt thereof, can be used in combination with a vaccine to stimulate an immune response against pathogens, toxins, and self-antigens.

[0145] When two or more agents are administered to a patient, they may be administered simultaneously, separately, sequentially, or in combination (eg, for three or more agents).

[0146] Formulations, dosage forms and routes of administration When used as a pharmaceutical, the compounds of the present disclosure can be administered in the form of a pharmaceutical composition. Accordingly, the present disclosure provides compositions comprising a compound of any of the formulas described herein, any of the compounds listed in any of the claims and described herein, or a pharmaceutically acceptable salt thereof, or any of the embodiments thereof, and at least one pharmaceutically acceptable carrier or excipient. These compositions can be prepared by methods well known in the pharmaceutical arts and can be administered by various routes, depending on whether local or systemic treatment is required and the area to be treated. Administration can be topical (including transdermal, epidermal, ocular, and mucosal delivery, including intranasal, vaginal, and rectal delivery), pulmonary (e.g., inhalation or insufflation of powder or aerosol by nebulizer, intratracheal, or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, or injection or infusion, or intracranial, e.g., intrathecal or intraventricular, administration.

[0147] Parenteral administration may be in the form of a single bolus dose or may be, for example, administered by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder, or oily bases, thickeners, and the like may be necessary or desirable.

[0148] The present application also includes pharmaceutical compositions comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof as an active ingredient in combination with one or more pharmaceutically acceptable carriers or excipients. In some embodiments, the compositions are suitable for topical administration. When preparing the compositions of the present application, the active ingredient is typically mixed with an excipient, diluted by an excipient, or enclosed within a carrier in the form of, for example, a capsule, sachet, paper, or other container. When an excipient serves as a diluent, it can be a solid, semi-solid, or liquid material and serves as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solids or in liquid media), ointments containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

[0149] When preparing formulation, active compound can be milled to provide suitable particle size before being combined with other components.If active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh.If active compound is substantially water-soluble, particle size can be adjusted by milling to provide substantially uniform distribution (for example, about 40 mesh) in formulation.

[0150] To obtain a particle size suitable for tablet formation and other formulation types, the compounds of the present application can be milled using known milling procedures, such as wet milling. Finely divided (nanoparticulate) preparations of the compounds of the present application can be prepared by processes well known in the art (see, for example, WO 2002 / 000196).

[0151] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulations may further include lubricants such as talc, magnesium stearate, and mineral oil, wetting agents, emulsifying and suspending agents, preservatives such as methyl benzoate and propyl hydroxybenzoate, sweeteners, and flavoring agents. The compositions of the present application can be formulated to provide rapid, sustained, or delayed release of the active ingredient after administration to a patient by using procedures well known in the art.

[0152] In some embodiments, the pharmaceutical composition comprises silicified microcrystalline cellulose (SMCC) and at least one compound described herein, or a pharmaceutically acceptable salt thereof. In some embodiments, the silicified microcrystalline cellulose comprises about 98% microcrystalline cellulose and about 2% silicon dioxide (w / w).

[0153] In some embodiments, the composition is a sustained-release composition comprising at least one compound described herein or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier or excipient. In some embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof and at least one component selected from microcrystalline cellulose, lactose monohydrate, hydroxypropyl methylcellulose, and polyethylene oxide. In some embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof, and microcrystalline cellulose, lactose monohydrate, and hydroxypropyl methylcellulose. In some embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof, and microcrystalline cellulose, lactose monohydrate, and polyethylene oxide. In some embodiments, the composition further comprises magnesium stearate or silicon dioxide. In some embodiments, the microcrystalline cellulose is Avicel PH102™. In some embodiments, the lactose monohydrate is Fast-flo 316™. In some embodiments, the hydroxypropyl methylcellulose is hydroxypropyl methylcellulose 2208 K4M (e.g., Methocel K4 M Premier™) and / or hydroxypropyl methylcellulose 2208 K100LV (e.g., Methocel K00LV™). In some embodiments, the polyethylene oxide is polyethylene oxide WSR1105 (e.g., Poly ox WSR 1105™).

[0154] In some embodiments, the compositions are manufactured using a wet granulation process. In some embodiments, the compositions are manufactured using a dry granulation process.

[0155] The compositions can be formulated in unit dosage form, each dosage containing from about 5 to about 1,000 mg (1 g), more usually about 100 mg to about 500 mg, of the active ingredient. In some embodiments, each dosage contains about 10 mg of the active ingredient. In some embodiments, each dosage contains about 50 mg of the active ingredient. In some embodiments, each dosage contains about 25 mg of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable as unitary administration for human subjects and other mammals, each unit containing a predetermined quantity of the active material calculated to produce the desired therapeutic effect, together with suitable pharmaceutical excipients.

[0156] The ingredients used to formulate pharmaceutical compositions are of high purity and substantially free of potentially harmful contaminants (e.g., at least national food grade, generally at least analytical grade, and more typically at least pharmaceutical grade). For human consumption in particular, compositions are preferably manufactured or formulated in accordance with good manufacturing practice as defined in applicable regulations of the U.S. Food and Drug Administration. For example, suitable formulations may be sterile and / or substantially isotonic and / or in full compliance with all U.S. Food and Drug Administration good manufacturing practice regulations.

[0157] The active compound may be effective over a wide dosage range and will generally be administered in a therapeutically effective amount. However, it will be understood that the amount of compound actually administered will usually be determined by the physician according to the relevant circumstances, including the condition to be treated, the selected route of administration, the compound actually administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, etc.

[0158] Therapeutic dosages of the compounds of the present application may vary depending, for example, on the particular application for which the treatment is given, the method of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compound of the present application in a pharmaceutical composition may vary depending on many factors, including dosage, chemical characteristics (e.g., hydrophobicity), and the route of administration. For example, the compounds of the present application may be provided in an aqueous physiological buffer solution containing about 0.1 to about 10% w / v of the compound for parenteral administration. Some typical dosage ranges are about 1 pg to about 1 g per kg of body weight per day. In some embodiments, the dosage range is about 0.01 mg to about 100 mg per kg of body weight per day. The dosage may depend on variables such as the type and progression of the disease or disorder, the overall health of the particular patient, the relative biological availability of the selected compound, the excipient formulation, and its route of administration. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0159] To prepare solid compositions such as tablets, the primary active ingredient is mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of the compounds of the present application. When these preformulation compositions are referred to as homogeneous, the active ingredient is typically dispersed evenly throughout the composition so that the composition can be readily subdivided into similarly effective unit dosage forms, such as tablets, pills, and capsules. This solid preformulation is then subdivided into unit dosage forms of the type described above, containing, for example, from about 0.1 to about 1000 mg of the active ingredient of the present application.

[0160] The tablets or pills of the present application can be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which resists disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, including many polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0161] Liquid forms into which the compounds and compositions of the present application can be incorporated for oral or injectable administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions including edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0162] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. The compositions can be nebulized using an inert gas. Nebulized solutions can be inhaled directly from the nebulizing device, or the nebulizing device can be attached to a face mask, tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions can be administered orally or nasally from devices that deliver the formulation in an appropriate manner.

[0163] Topical formulations can include one or more conventional carriers. In some embodiments, ointments can include water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, etc. Cream carrier compositions can be based on water in combination with glycerol and one or more other ingredients, such as glyceryl monostearate, PEG-glyceryl monostearate, and cetylstearyl alcohol. Gels can be formulated using isopropyl alcohol and water, appropriately combined with other ingredients such as glycerol and hydroxyethylcellulose. In some embodiments, topical formulations include at least about 0.1, at least about 0.25, at least about 0.5, at least about 1, at least about 2, or at least about 5% by weight of a compound of the present application. Topical formulations can be suitably packaged, for example, in 100g tubes, optionally accompanied by instructions for the treatment of a selected indication, such as psoriasis or other skin conditions.

[0164] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of the administration, such as prophylaxis or treatment, the condition of the patient, the method of administration, etc. In therapeutic applications, compositions can be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will depend on the disease state being treated and the judgment of the attending physician, depending on factors such as the severity of the disease, the age, weight, and general condition of the patient, etc.

[0165] The compositions administered to patients may be in the form of pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or may be sterile filtered. Aqueous solutions may be packaged for immediate use or lyophilized, with the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the compound formulations is typically 3-11, more preferably 5-9, and most preferably 7-8. It will be understood that the use of certain of the aforementioned excipients, carriers, or stabilizers may result in the formation of pharmaceutical salts.

[0166] Therapeutic dosages of the compounds of the present application may vary depending, for example, on the particular application for which the treatment is given, the method of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compound of the present application in a pharmaceutical composition may vary depending on many factors, including dosage, chemical characteristics (e.g., hydrophobicity), and the route of administration. For example, the compounds of the present application may be provided in an aqueous physiological buffer solution containing about 0.1 to about 10% w / v of the compound for parenteral administration. Some typical dosage ranges are about 1 pg to about 1 g per kg of body weight per day. In some embodiments, the dosage range is about 0.01 mg to about 100 mg per kg of body weight per day. The dosage may depend on variables such as the type and progression of the disease or disorder, the overall health of the particular patient, the relative biological availability of the selected compound, the excipient formulation, and its route of administration. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0167] IV.Synthesis method The compounds of the present application (including salts thereof) may be prepared using known organic synthesis techniques, or may be synthesized according to any of a number of possible synthetic routes.

[0168] The reactions for preparing the compounds of the present application can be carried out in a suitable solvent that can be easily selected by one skilled in the art of organic synthesis. A suitable solvent can be substantially non-reactive with the starting materials (reactants), intermediates, or products at the temperature at which the reaction is carried out, for example, at a temperature ranging from the freezing temperature of the solvent to the boiling point of the solvent. A given reaction can be carried out in one solvent or a mixture of two or more solvents. Depending on the specific reaction step, one skilled in the art can select a solvent suitable for the particular reaction step.

[0169] The preparation of the compounds of the present application can involve the protection and deprotection of various chemical groups. The need for protection and deprotection, and the selection of appropriate protecting groups, can be easily determined by one skilled in the art. Protecting group chemistry is described, for example, in Kocienski, Protecting Groups, (Thieme, 2007), Robertson, Protecting Group Chemistry, (Oxford University Press, 2000), Smith et al., March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 6th Ed. (Wiley, 2007), Peturssion et al., "Protecting Groups in Carbohydrate Chemistry," J. Chem. Educ., 1997, 77(11), 1297, and Wuts et al., Protective Groups in Organic Synthesis, 4th Ed. (Wiley, 2006).

[0170] Reactions can be monitored according to any suitable method known in the art, for example, product formation can be monitored by spectroscopic means such as nuclear magnetic resonance spectroscopy (e.g., 1H or 13C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), mass spectrometry, or by chromatographic methods such as high performance liquid chromatography (HPLC) or thin layer chromatography (TLC).

[0171] The present application is further illustrated by the following examples, which should not be construed as limiting. The contents of all references, patents, and published patent applications, as well as figures and tables, cited throughout this application are hereby incorporated by reference. [Example]

[0172] Chemical synthesis examples Example 1 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-((5-oxopyrrolidin-2-yl)methyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 1) [ka] Compound 1D-1 (1.00 g, 8.7 mmol, 1.0 equivalent) and trimethylamine (1.76 g, 17.37 mmol, 2.0 equivalents) were dissolved in DCM (20 mL). 4-Methylbenzenesulfonyl chloride (1.99 g, 10.42 mmol, 1.20 equivalents) was added at 0 °C and stirred for 1 hour. The mixture was concentrated and purified by column chromatography to give compound 1D (2.1 g, yield: 89.8%).

[0173] [ka] In a 500 mL round-bottom flask, 3-bromo-2-chlorophenol (12.4 g, 0.060 mol, 1.0 equiv.), B2Pin2 (16.4 g, 0.065 mol, 1.08 equiv.), KOAc (20.5 g, 0.210 mol, 3.5 equiv.), and Pd(dppf)Cl 2· DCM (4.1 g, 5.1 mmol, 0.085 equiv) was added, followed by dioxane (300 mL), and the final mixture was backfilled with N and stirred for 3 h at 95° C. The reaction mixture was then cooled to room temperature, filtered, the filter cake was washed with dioxane (100 mL), and the filtrate was used directly in the next step.

[0174] To the previous filtrate, 3-bromo-2-chlorophenol (12.0 g, 0.059 mol, 0.99 equiv.), K2CO3 (24.8 g, 0.180 mol, 3.0 equiv.), and Pd(dppf)Cl2·DCM (2.1 g, 2.40 mmol, 0.042 equiv.) were added, followed by HO (80 mL). The final mixture was backfilled with N2 and stirred at 85 °C for 3.5 h. The reaction mixture was then cooled to room temperature, filtered, and the filter cake was washed with EtOAc (300 mL). Brine (300 mL) was added to the filtrate, and the layers were separated. The aqueous phase was extracted with EtOAc (100 mL × 2). The combined organic phase was decolorized with activated charcoal at room temperature overnight. The mixture was filtered through a pad of Celite, the filter cake was washed with EtOAc, and the combined organic phase was concentrated in vacuo. The residue was purified by recrystallization from DCM / PE=1.5 / 1 to give the desired product SM1-2A (10.1 g, yield: 66%).

[0175] [ka] To a stirred mixture of SM1-2A (10.1 g, 0.039 mol, 1.0 equiv.) in DCM (200 mL) was added DIPEA (19.4 g, 0.151 mol, 3.8 equiv.) at 0 °C, followed by TfO (26.8 g, 0.095 mol, 2.4 equiv.) at this temperature, and the mixture was stirred at room temperature for an additional 2 h. Water (100 mL) was added, and the layers were separated. The aqueous phase was extracted with DCM (100 mL). The combined organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by recrystallization from EtOH / HO = 1 / 1 to give the desired product SM1-3A (18.3 g, 89% yield).

[0176] [ka] To a stirred solution of SM1-3A (14.2 g, 0.027 mol, 1.0 equiv) in dioxane (80 mL), B2Pin2 (27.8 g, 0.109 mol, 4.0 equiv), KOAc (16 g, 0.164 mol, 6.0 equiv), and Pd(dppf)Cl₂·DCM (3.3 g, 4.1 mmol, 0.15 equiv) were added at room temperature, followed by a nitrogen atmosphere and stirring at 85 °C for 2 h. After cooling to room temperature, EtOAc (150 mL) and water (150 mL) were added to the mixture, which was then separated. The aqueous phase was extracted with EtOAc (100 mL). The combined organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was dissolved in 50 mL of EtOAc, and then 300 mL of PE was slowly added to the solution to form a black suspension. After stirring for 30 minutes, it was filtered, and the filter cake was washed with 140 mL of 6 / 1 PE / EtOAc. The filtrate was concentrated in vacuo, and the residue was recrystallized with 150 mL of EtOH to give the desired product 1A (9.5 g, 75% yield) as an off-white solid.

[0177] [ka] Compound 1A (1.0 g, 2.1 mmol, 1.0 equiv.), compound 1B (1.138 g, 5.32 mmol, 2.5 equiv.), K2CO3 (1.45 g, 10.5 mmol, 5.0 equiv.), and Pd(dppf)Cl2·DCM (0.52 g, 0.63 mmol, 0.3 equiv.) were dissolved in DMF (15 mL) and HO (3 mL) under a N2 atmosphere. The reaction mixture was stirred at 100 °C for 3 h. After cooling, 30 mL of water and 30 mL of EtOAc were added. The organic phase was washed with water and concentrated to give a residue, which was then washed with EtOAc to give compound 1C (0.80 g, 77.8% yield). LCMS(ESI):C 24 H 14 Calculated for Cl2N6O2; [M+H]+: 489.1, Found: 489.1. 1H NMR (500 MHz, DMSO-d6) δ 8.11 (s, 2H), 7.97 (d, J = 1.9 Hz, 2H), 7.71 (dd, J = 7.9, 1.7 Hz, 2H), 7.48 (t, J = 7.6 Hz, 2H), 7.32 (dd, J = 7.6, 1.7 Hz, 2H), 7.25 (d, J = 1.9 Hz, 2H), 6.31 (s, 2H).

[0178] [ka] Compound 1C (100 mg, 0.20 mmol, 1.0 equiv.), compound 1D (121 mg, 0.45 mmol, 2.2 equiv.), and CsCO (333 mg, 1.02 mmol, 5.0 equiv.) were dissolved in DMF (2.0 mL). The reaction mixture was stirred at 60 °C for 1 h. After cooling, 10 mL of water was added, extracted with EtOAc, and concentrated. It was purified by preparative HPLC to give compound 1 (80 mg, yield: 57.3%). LCMS(ESI):C 34 H 28 Calculated for Cl2N8O4; [M+H]+: 683.2, Found: 683.2. 1 H NMR (500 MHz, DMSO-d6) δ 8.11 (s, 2H), 8.01 (d, J = 1.2Hz, 2H), 7.87 (s, 2H), 7.70 (d, J = 6.4 Hz, 2H), 7.49 (t, J = 6.4 Hz, 2H), 7.32 (d, J = 6 Hz, 2H), 7.26 (d, J = 1.6 Hz, 2H), 3.88-3.94 (m, 6H), 2.21-2.27 (m, 2H), 2.05-2.18 (m, 4H), 1.72-1.77 (m, 2H).

[0179] Example 2 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(((S)-5-oxopyrrolidin-2-yl)methyl)thieno[2,3-d]pyrimidin-4(3H)-one) (Compound 2) [ka] Following the procedure for compound 1, the title compound 2 was obtained by substituting 6-bromothieno[2,3-d]pyrimidin-4(3H)-one for 1B. LCMS(ESI):C 34 H 26 Calculated for Cl2N6O4S2; [M+H]+: 717.1, 719.1, Found: 717.1, 719.1. 1H NMR (500 MHz, DMSO-d6): δ 8.43 (s, 2H), 7.85 (s, 2H), 7.80 (d, J = 7.7 Hz, 2H), 7.70 (s, 2H), 7.61-7.55 (m, 3H), 7.50 (d, J = 7.4 Hz, 2H), 4.06-4.01 (m, 4H), 4.01-3.96 (m, 2H), 2.29-2.22 (m, 2H), 2.18-2.09 (m, 4H), 1.81-1.73 (m, 2H).

[0180] Example 3 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(7-fluoro-3-((5-oxopyrrolidin-2-yl)methyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 3) [ka] Compound 1B (1.0 g, 4.6 mmol, 1.0 equiv.) and Selectfluor (2.48 g, 7.0 mmol, 1.5 equiv.) were dissolved in CHCN (16 mL) and DMF (4.0 mL). The reaction was carried out at 50 °C for 2 h. Compound 3A (0.30 g, 27.7% yield) was obtained by precipitation with 40 mL of water. LCMS (ESI): Calculated for C6H3BrFN3O; [M+H]+: 231.9, Found: 231.9.

[0181] Following the procedure for compound 1, the title compound 3 was obtained by substituting 6-bromo-7-fluoropyrrolo[2,1-f][1,2,4]triazin-4(1H)-one (3A) for 1B. LCMS(ESI):C 34 H 26 Calculated for Cl2F2N8O4; [M+H]+: 719.1721.1, Found: 719.1, 721.1. 1 H NMR (500 Mhz, DMSO-d6): δ 8.01 (d, J = 1.2 Hz, 2H), 7.87 (s, 2H), 7.72 (d, J = 6.4 Hz, 2H), 7.49 (t, J = 6.4 Hz, 2H), 7.32 (d, J = 6 Hz,2H), 7.26 (d, J = 1.6 Hz, 2H), 3.94-3.88 (m, 6H), 2.27-2.22 (m, 2H), 2.18-2.10 (m, 4H), 1.78-1.77 (m, 2H).

[0182] Example 4 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(5-methyl-3-(((S)-5-oxopyrrolidin-2-yl)methyl)-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one) (Compound 4) [ka] Following the procedure for compound 1, substituting 6-bromo-5-methyl-3,5-dihydro-4H-pyrrolo[3,2-d]pyrimidin-4-one for 1B, the title compound 4 was obtained. LCMS(ESI):C 36 H 32 Calculated for Cl2N8O4; [M+H]+: 711.2, 713.2, Found: 711.2, 713.2. 1H NMR (500 MHz, DMSO-d6) δ 8.10 (d, J = 2.0 Hz, 2H), 7.83-7.79 (m, 4H), 7.48 (t, J = 7.5 Hz, 2H), 7.29 (d, J = 7.5 Hz, 2H), 4.08 (s, 6H), 3.99-3.95 (m, 6H), 2.23-2.19 (m, 2H), 2.12-2.04 (m, 2H), 2.02-1.96 (m, 2H), 1.79-1.76 (m, 2H).

[0183] Example 5 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 5) [ka] Following the procedure for compound 1, the title compound 5 was obtained by substituting 7-bromopyrrolo[1,2-a]pyrazin-1(2H)-one for 1B. LCMS(ESI):C 36 H 30 Calculated for Cl2N6O4; [M+H]+: 681.2, Found: 683.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.87 (s, 2H), 7.79 (s, 2H), 7.71-7.65 (m, 2H), 7.51-7.42 (m, 4H), 7.33-7.27 (m, 2H), 7.25 (s, 2H), 6.87 (d, J = 5.9 Hz, 2H), 4.02-3.94 (m, 3H), 3.76-3.69 (m, 3H), 2.23-2.14 (m, 2H), 2.13-2.02 (m, 4H), 1.81-1.72 (m, 2H).

[0184] Example 6 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(1-methyl-5-(((S)-5-oxopyrrolidin-2-yl)methyl)-1,5-dihydro-4H-imidazo[4,5-c]pyridin-4-one) (Compound 6) [ka] Following the procedure for compound 1, the title compound 6 was obtained by substituting 2-bromo-1-methyl-1,5-dihydro-4H-imidazo[4,5-c]pyridin-4-one for 1B. LCMS(ESI):C 36 H 32 Calculated for Cl2N8O4; [M+H]+: 711.2, 713.2, Found: 711.2, 713.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.87 (s, 2H), 7.79 (s, 2H), 7.71-7.65 (m, 2H), 7.51-7.42 (m, 4H), 7.33-7.27 (m, 2H), 7.25 (s, 2H), 6.87 (d, J = 5.9 Hz, 2H), 4.02-3.94 (m, 3H), 3.76-3.69 (m, 3H), 2.23-2.14 (m, 2H), 2.13-2.02 (m, 4H), 1.81-1.72 (m, 2H).

[0185] Example 7 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(5-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 7) [ka] Following the procedure for compound 1, substituting 2-bromopyrazolo[1,5-a]pyrazin-4(5H)-one for 1B gave the title compound 7. LCMS(ESI):C 34 H 28Calculated for Cl2N8O4; [M+H]+: 683.2, 685.2, Found: 683.2, 685.2. 1 H NMR (500 MHz, DMSO-d6) δ 7.89-7.87 (m, 4H), 7.80 (s, 2H), 7.57 (t, J = 10 Hz, 2H), 7.48-7.46 (s, 2H), 7.40 (s, 2H), 7.20 (t, J = 4.8Hz, 2H), 4.05-3.97 (m, 4H), 3.87-3.83 (m, 2H), 2.26-2.21 (m, 2H), 2.14-2.07 (m, 4H), 1.79-1.74 (m, 2H).

[0186] Example 8 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(5-(((S)-5-oxopyrrolidin-2-yl)methyl)thieno[3,2-c]pyridin-4(5H)-one) (Compound 8) [ka] Following the procedure for compound 1, substituting 2-bromothieno[3,2-c]pyridin-4(5H)-one for 1B gave the title compound 8. LCMS(ESI):C 36 H 28 Calculated for Cl2N4O4S2; [M+H]+: 715.1, 717.1, Found: 715.1, 717.1. 1 H NMR (500 MHz, DMSO-d6) δ 7.80-7.76 (m, 6H), 7.61-7.54 (m, 4H), 7.47 (dd, J = 7.6, 1.6 Hz, 2H), 6.97 (d, J = 7.2 Hz, 2H), 4.15-4.09 (m, 2H), 4.00-3.87 (m, 4H), 2.26-2.12 (m, 2H), 2.11-1.89 (m, 4H), 1.85-1.70 (m, 2H).

[0187] Example 9 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-(((S)-5-oxopyrrolidin-2-yl)methyl)-3,4-dihydropyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 9) [ka] Following the procedure for compound 1, substituting 7-bromo-3,4-dihydropyrrolo[1,2-a]pyrazin-1(2H)-one for 1B gave the title compound 9. LCMS(ESI):C 36 H 34 Calculated for Cl2N6O4; [M+H]+: 684.2, 686.2, Found: 684.2, 686.2. 1 H NMR (500 MHz, DMSO-d6) δ 7.77 (s, 2H), 7.58 (d, J = 6 Hz, 2H), 7.47 (s, 2H), 7.41 (t, J = 6 Hz, 2H), 7.20 (d, J = 6 Hz, 2H), 7.00 (s, 2H), 4.15 (t, J = 4.4 Hz, 4H), 3.53 (s, 4H).

[0188] Example 10 Preparation of 8,8'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3,7-dimethyl-1-(((S)-5-oxopyrrolidine)-2-yl)methyl)-3,7-dihydro-1H-purine-2,6-dione) (Compound 10) [ka] Following the procedure for compound 1, substituting 8-bromo-3,7-dimethyl-3,7-dihydro-1H-purine-2,6-dione for 1B, gave the title compound 10. LCMS(ESI):C 36 H 34 Cl2N 10 Calculated values ​​for O6: [M+H]+: 772.2, 774.2, Found: 772.2, 774.2. 1H NMR (500 MHz, DMSO-d6) δ 7.81 (d, J = 5.0 Hz, 2H), 7.68 (d, J = 5.0 Hz, 2H), 7.52 (t, J = 10.0 Hz, 2H), 7.12 (d, J = 5.0 Hz, 2H), 3.98-3.92 (m, 6H), 3.58 (s, 6H), 3.32 (s, 6H), 2.38 -2.26 (m, 2H), 2.14-2.09 (m, 4H), 1.83-1.76 (m, 2H).

[0189] Example 11 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrrolo[1,2-d][1,2,4]triazin-1(2H)-one) (Compound 11) [ka] Following the procedure for compound 1, substituting 8-bromo-3,7-dimethyl-3,7-dihydro-1H-purine-2,6-dione for 1B, gave the title compound 11. LCMS(ESI):C 34 H 28 Calculated for Cl2N8O4; [M+H]+: 682.1, 684.1, Found: 682.1, 684.1. 1 H 1 H NMR (500 MHz, DMSO-d6) δ 8.36 (s, 2H), 7.92 (d, J = 1.8 Hz, 2H), 7.71-7.63 (m, 6H), 7.54-7.48 (m, 2H), 7.30-7.24 (m, 2H), 4.25-4.09 (m, 6H), 2.40-2.26 (m, 4H), 1.99-1.77 (m, 4H).

[0190] Example 12 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(7-(((S)-5-oxopyrrolidin-2-yl)methyl)imidazo[1,2-a]pyrazin-8(7H)-one) (Compound 12) [ka] To a stirred solution of compound 12A (770 mg, 3.53 mmol, 1 equiv.) in DMF (10 mL) was added CsCO (2302 mg, 7.06 mmol, 2.0 equiv.) and 2-bromo-1,1-dimethoxyethane (891 mg, 5.30 mmol, 1.5 equiv.) at 25 °C. The resulting mixture was stirred at 80 °C for 16 h. After cooling to 25 °C, the reaction was quenched with water (10 mL) and extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAC (5:1 to 3:1) to give compound 12B (451 mg, yield: 41.7%).

[0191] To a stirred solution of compound 12B (451 mg, 1.47 mmol, 1.0 equiv) in EtOH / HO (5 mL / 1 mL) was added NaOH (89 mg, 2.22 mmol, 1.5 equiv) at 25 °C. The resulting mixture was stirred at 80 °C for 2 h. After cooling to 25 °C, the reaction was quenched with HOAc solution (1 M in water) and adjusted to pH 6.0. It was extracted with EtOAc (30 mL × 2), and the combined organic layer was washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give compound 12C (410 mg, yield: 100%).

[0192] To a stirred solution of compound 12C (410 mg, 1.47 mmol, 1.0 equiv) in DMF (5 mL) was added NH4Cl (786 mg, 14.7 mmol, 10 equiv), DIEA (508 mg, 4.41 mmol, 3.0 equiv), and HATU (1040 mg, 2.94 mmol, 2 equiv) at 25 °C. The resulting mixture was stirred at 25 °C for 16 h. The reaction was quenched with water (100 mL) and extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1 to 2:1) to give compound 12D (300 mg, yield: 66.8%). LCMS (ESI): CH 12 Calculated for BrN3O3; [M+H]+: 277.01, 279.00, Found: 246.11, 248.09.

[0193] Compound 12D (300 mg, 1.08 mmol, 1.0 equiv) was dissolved in HOAc (3.0 mL) at 25 °C, and the resulting mixture was stirred at 105 °C for 4 h. After the reaction was completed, the mixture was concentrated under reduced pressure to give compound 12E (230 mg, 100% yield). LCMS (ESI): calculated for CHBrNO; [M+H]: 214.02, 216.02, found: 214.09, 216.07.

[0194] Following the procedure for compound 1, replacing 1B with 12E gave the title compound 12. LCMS (ESI): C 34 H 28 Calculated for Cl2N8O4; [M+H]+: 682.2, 684.2, Found: 682.2, 684.2. 1 H NMR (500 Mhz, DMSO-d6) δ 1H NMR (500 Mhz, DMSO-d6) 8.43 (d, J = 2.0 Hz, 2H), 8.23 ​​(d, J = 7.9 Hz, 2H), 7.79 (d, J = 6.5 Hz, 2H), 7.63 (d, J = 5.7 Hz, 2H), 7.57 (t, J = 7.7 Hz, 2H), 7.38 (d, J = 7.5 Hz, 2H), 7.15 (dd, J = 5.9, 1.7 Hz, 2H), 4.04-4.02 (m, 1H), 4.00-3.97 (m, 2H), 3.87-3.84 (m, 2H), 3.84-3.81 (m, 1H), 2.31-2.18 (m, 3H), 2.13-2.08 (m, 3H), 1.81-1.74 (m, 2H).

[0195] Example 13 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-fluoro-7-(((S)-5-oxopyrrolidin-2-yl)methyl)imidazo[1,2-a]pyrazin-8(7H)-one) (Compound 13) [ka] A mixture of 2-bromo-1,1-dimethoxyethane (5.8 mL, 49.3 mmol, 4.2 equiv) and HBr (1.4 mL, 48% in water, 12.5 mol, 1.08 equiv) was stirred at 100 °C. After 2 h, the mixture was cooled to room temperature and slowly added dropwise to a stirred solution of NaHCO in propan-2-ol (30 mL). A white solid separated. After filtration, 13A (1.5 g, 11.6 mmol, 1.0 equiv) was added to the filtrate, and the reaction was stirred at 100 °C for 2 h. After cooling, the mixture was filtered to obtain a cake, which was dissolved in DCM (50 mL), washed with saturated sodium bicarbonate solution (50 mL × 2), and dried over anhydrous magnesium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The mixture was concentrated under reduced pressure to give compound 13B (1.5 g, yield: 84%). LCMS (ESI): Calculated for C6H4ClN3; [M+H]+: 154.01, Found: 154.23

[0196] To a solution of compound 13B (1.5 g, 9.8 mmol, 1.0 equiv.) in MeCN (20 mL) was added Selectfluor (3.47 g, 9.8 mmol, 1.0 equiv.) at 25 °C, and the reaction was stirred at 70 °C for 1 h. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1 to 3:1) to give 13C (0.56 g, 33.5% yield). LCMS (ESI): calculated for CHClFN; [M+H]: 171.56, found: 172.10

[0197] To a solution of compound 13C (363 mg, 2.12 mmol, 1 equiv.) in THF (10 mL) was added n-BuLi (1.56 mL, 1.6 M in THF, 2.54 mmol, 1.2 equiv.) at -60 °C, and the reaction was stirred at -60 °C for 30 min. Br2 (0.16 mL, 3.18 mmol, 1.5 equiv.) was then added to the mixture at -60 °C and stirred for 2 h. The mixture was quenched with saturated NH4Cl solution (3.0 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1 to 3:1) to give 13D (183 mg, 34.6% yield). LCMS (ESI): calculated for CH2BrClFN3; [M+H]+: 250.46, found: 250.32.

[0198] Compound 13D (83 mg, 0.33 mmol, 1.0 equiv) was dissolved in HCOOH (2.0 mL) at 25 °C, and the reaction was stirred at 110 °C for 1.5 h. After completion of the reaction, the mixture was concentrated under reduced pressure to give 13E (76 mg, 100% yield). LCMS (ESI): Calcd for CHBrFN0; [M+H]: 234.9, Found: 234.1

[0199] Following the procedure for compound 1, replacing 1B with 13E gave the title compound 13. LCMS (ESI): C 34 H 26 Calculated for Cl2F2N8O4; [M+H]+: 719.5, 721.5, Found: 719.5, 721.5. 1H NMR (500 MHz, DMSO-d6) δ 8.42 (d, J = 2.0 Hz, 2H), 8.22 (d, J = 7.9 Hz, 2H), 7.62 (d, J = 5.7 Hz, 2H), 7.56 (t, J = 7.7 Hz, 2H), 7.37 (d, J = 7.5 Hz, 2H), 7.14 (dd, J = 5.9, 1.7 Hz, 2H), 4.03 - 3.81 (m, 6H), 2.32-2.19 (m, 3H), 2.14-2.09 (m, 3H), 1.80-1.73 (m, 2H).

[0200] Example 14 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(((S)-5-oxopyrrolidin-2-yl)methyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one) (Compound 14) [ka] Following the procedure for compound 1, replacing 1B with 6-bromo-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one gave the title compound 14. LCMS (ESI): C 34 H 28 Calculated for Cl2N8O4; [M+H]+: 683.5, 685.5, Found: 683.5, 685.5. 1 H NMR (500 MHz, DMSO-d6) δ 12.40 (s, 2H), 8.17 (s, 2H), 7.83 (s, 2H), 7.71 (d, J = 2.5 Hz, 2H), 7.54 (t, J = 5.0 Hz, 2H), 7.38 (d, J = 5.0 Hz, 2H), 6.89 (d, J = 2.5 Hz, 2H), 4.08- 4.01 (m, 2H), 3.98-3.93 (m, 4H), 2.24-2.17 (m, 2H), 2.10-2.03 (m, 4H), 1.79-1.74 (m, 2H).

[0201] Example 15 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(7-(((S)-5-oxopyrrolidin-2-yl)methyl)-[1,2,4]triazolo[1,5-a]pyrazin-8(7H)-one) (Compound 15) [ka] To a solution of 3-chloropyrazin-2-amine (1.0 g, 7.78 mmol, 1.0 equiv) in dioxane (100 mL), isothiocyanato-O-ethylcarboisothiocyanatidate (1.18 g, 7.9 mmol, 1.02 equiv) was added dropwise and stirred at room temperature for 24 h. The solvent was evaporated, and the residue was purified by flash chromatography on silica gel using a gradient of 0% to 5% MeOH in DCM. The title compound 15B was obtained as a yellow solid (1.13 g, 56%). LCMS (ESI): Calculated for C8H9ClN4O2S; [M+H]+: 261.0, 263.0, Found: 261.0, 263.0.

[0202] Compound 15B (0.73 g, 2.98 mmol, 1.0 equiv.), hydroxylamine hydrochloride (1.03 g, 74.2 mmol, 25 equiv.), and diisopropylethylamine (1.15 g, 1.55 mL, 8.9 mmol, 3.0 equiv.) were dissolved in MeOH (4.0 mL) and EtOH (4.0 mL). The reaction mixture was heated to 60 °C for 3 h. The solvent was evaporated, and the residue was suspended in DCM (30 mL) and water (6.0 mL). The suspension was stirred for 10 min, and the solid was removed by filtration. The aqueous phase was extracted four times with DCM, and the combined organic layers were dried over NaSO, and the solvent was removed in vacuo. The residue was combined with the filtered solid compound 15C (808 mg, 170% yield) and used in the next step without purification. LCMS (ESI): Calculated for C5H4ClN5; [M+H]+: 170.0, 172.0, Found: 170.0, 172.0.

[0203] Cuprous bromide (1.06 g, 4.8 mmol), tert-butyl nitrite (505 mg, 0.605 mL, 4.8 mmol), and compound 15C were dissolved in acetonitrile (60 mL). The reaction mixture was heated to 75 °C for 35 min. After cooling, saturated NaHCO solution (20 mL) was added, separated, and extracted with EtOAc (40 mL). The combined organic layers were then dried over NaSO, and the solvent was removed in vacuo. The crude material was purified by flash chromatography on silica gel (0% to 50% EtOAc in heptane) to give an off-white solid, 15D (202 mg, 29%). LCMS (ESI): calculated for CHBrClN; [M+H]: 232.9, 234.9, 236.9, found: 232.9, 234.9, 236.9.

[0204] Compound 15D (100 mg, 0.465 mmol, 1.0 equiv) was dissolved in 5.0 mL of formic acid and heated to 100 °C for 8 h. The reaction solution was cooled, concentrated, and the solid precipitated to give compound 15E (102 mg, 55.9%). LCMS (ESI): Calcd for CHBrNO; [M+H]: 215.0, 217.0, Found: 215.0, 217.0.

[0205] Following the procedure for compound 1, substituting 15E for 1B gave the title compound 15. LCMS (ESI): C 32 H 26 Cl2N 10 Calculated values ​​for O4: [M+H]+: 685.5, 687.5, Found: 685.5, 687.5. 1H NMR (500 MHz, DMSO-d6) δ 8.09 (d, J = 5.0 Hz, 2H), 7.95 (d, J = 5.0 Hz, 2H), 7.75 (s, 2H), 7.63 (t, J = 7.5 Hz, 2H), 7.58 (t, J = 5.0 Hz, 2H), 7.45 (d, J = 10.0 Hz, 2H), 4.13- 4.01 (m, 4H), 3.99-3.93 (m, 2H), 2.32-2.26 (m, 2H), 2.19-2.07 (m, 4H), 1.80-1.74 (m, 2H).

[0206] Example 16 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(4-fluoro-2-((5-oxopyrrolidin-2-yl)methyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 16) [ka] Compound 16A (200 mg, 0.938 mmol, 1.0 equiv) and Selectfluor (300 mg, 0.75 mmol, 0.8 equiv) were dissolved in DCM (4.0 mL) and HO (4 mL) and stirred at 25 °C overnight. Water (20 mL) was added to the reaction mixture, which was then extracted with DCM (20 mL × 3). The combined organic layer was dried over NaSO, filtered, and concentrated under reduced pressure to give the crude product. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (3:1) to give compound 16B (110 mg, 49.4% yield). LCMS (ESI): calculated for CHBrFNO; [M+H]: 231.01, 233.01, found: 231.01, 233.01.

[0207] Following the procedure for compound 1, replacing 1B with 16B gave the title compound 16. LCMS (ESI): C 36 H 28 Calculated for Cl2F2N6O4; [M+H]+: 717.2, 719.2, Found: 717.2, 719.2. 1H NMR (500 MHz, DMSO-d6) δ 8.10 (d, J = 2.5 Hz, 2H), 7.87 (d, J = 5.0 Hz, 2H), 7.66 (d, J = 7.5 Hz, 2H), 7.45 (d, J = 5.0 Hz, 2H), 7.40 (d, t = 7.5 Hz, 2H), 7.32 (d, J = 2.5 Hz, 2H), 6.73 (d, J = 2.5 Hz, 2H), 3.93 - 3.81 (m, 6H), 2.24 - 2.19 (m, 2H), 2.12 - 2.04 (m, 4H), 1.77 - 1.69 (m, 2H).

[0208] Example 17 Preparation of 6,6'-(2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)bis(3-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 17) [ka] Following the procedure for compound 1, substituting 3-bromo-2-methylphenol for 3-bromo-2-chlorophenol, gave the title compound 17. LCMS (ESI): C 36 H 34 Calculated for N8O4; [M+H]+ 643.3, 645.3, Found: 643.3, 645.3. 1 H NMR (500 Mhz, DMSO-d6) δ 8.09 (s, 2H), 7.83-7.87 (m, 4H), 7.40-7.42 (m, 2H), 7.31 (t, J = 6.4 Hz), 7.09 (s, 2H), 3.89-3.93 (m, 6H), 2.18-2.26 (m, 2H), 2.04-2.11 (m, 10H), 1.73-1.77 (m, 2H).

[0209] Example 18 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-((5-oxopyrrolidin-3-yl)methyl)pyrrolo[2,1-f]][1,2,4]triazin-4(3H)-one) (Compound 18) [ka] Following the procedure for compound 1, replacing 1D-1 with 4-(hydroxymethyl)pyrrolidin-2-one gave the title compound 18. LCMS (ESI): C 34 H 28 Calculated for Cl2N8O4; [M+H]+: 683.2, 685.2, Found: 683.2, 685.2. 1 H NMR (500 Mhz, DMSO-d6) δ 8.61 (d, J = 1.6 Hz, 2H), 8.09 (s, 2H), 7.94 (d, J = 1.6 Hz, 2H), 7.68 (m, 4H), 7.52 (t, J = 7.6 Hz, 2H), 6.45 (m, 2H), 4.24-3.71 (m, 4H), 3.46 (m, 2H), 3.31 (m, 2H), 2.55-2.47 (m, 2H), 2.45 (m, 2H), 2.36 (m, 2H).

[0210] Example 19 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(2-(2-oxoimidazolidin-1-yl)ethyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 19) [ka] Following the procedure for compound 1, replacing 1D-1 with 1-(2-hydroxyethyl)imidazolidin-2-one gave the title compound 19. LCMS (ESI): C 34 H 30 Cl2N 10 Calculated values ​​for O4: [M+H]+: 712.2, 714.2, Found: 712.2, 714.2. 1 H NMR (500 Mhz, DMSO-d6) δ 8.11 (s, 2H), 7.97 (d, J = 1.9 Hz, 2H), 7.71 (dd, J = 7.9, 1.7 Hz, 2H), 7.48 (t, J = 7.6 Hz, 2H), 7.32 (dd, J = 7.6, 1.7 Hz, 2H), 7.25 (d, J = 1.9 Hz, 2H), 6.31 (s, 2H), 3.98 (t, J = 5.6 Hz, 4H), 3.45 (dd, J = 9.0, 6.7 Hz, 4H), 3.38 (t, J = 5.4 Hz, 4H), 3.21 (dd, J = 8.9, 6.8 Hz, 4H).

[0211] Example 20 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(2-hydroxyethyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 20) [ka] Following the procedure for compound 1, substituting 2-bromoethan-1-ol for 1D gave the title compound 20. LCMS (ESI): C 28 H 22 Calculated for Cl2N6O4; [M+H]+: 577.1, 579.1, Found: 577.1, 579.1. 1H NMR (500 Mhz, DMSO-d6) δ 8.09-7.98 (m, 3H), 7.91 (d, J = 4.0 Hz, 1H), 7.70 (d, J = 7.7 Hz, 2H), 7.48 (m, 2H), 7.38-7.21 (m, 4H), 3.93 (m, 4H), 3.63 (m, 4H).

[0212] Example 21 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(2-(1H-pyrazol-1-yl)ethyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 21) [ka] Following the procedure for compound 1, replacing 1D-1 with 2-(1H-pyrazol-1-yl)ethan-1-ol gave the title compound 21. LCMS (ESI): C 34 H 26 Cl2N 10 Calculated for O2: [M+H]+: 677.2, 679.2, Found: 677.2, 679.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.96 (d, J = 1.9 Hz, 2H), 7.74 (d, J = 2.2 Hz, 2H), 7.70 (dd, J = 7.7, 1.7 Hz, 2H), 7.52-7.46 (m, 4H), 7.43 (s, 2H), 7.32 (dd, J = 7.6, 1.6 Hz, 2H), 7.28 (d, J = 1.9 Hz, 2H), 6.26-6.22 (m, 2H), 4.46 (t, J = 5.7 Hz, 4H), 4.28 (t, J = 5.7 Hz, 4H).

[0213] Example 22 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(2-(1H-1,2,4-triazol-1)-yl)ethyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 22) [ka] Following the procedure for compound 1, replacing 1D-1 with 2-(1H-1,2,4-triazol-1-yl)ethan-1-ol gave the title compound 22. LCMS (ESI): C 32 H 24 Cl2N 12 Calculated for O2; [M+H]+: 679.2, 681.2, Found: 679.2, 681.2. 1H NMR (500 Mhz, DMSO-d6) δ 8.54 (s, 2H), 7.99 (t, J = 1.7 Hz, 4H), 7.73-7.66 (m, 4H), 7.48 (t, J = 7.6 Hz, 2H), 7.32 (dd, J = 7.5, 1.6 Hz, 2H), 7.28 (d, J = 1.7 Hz, 2H), 4.56 (d, J = 5.8 Hz, 4H), 4.29 (t, J = 5.7 Hz, 4H).

[0214] Example 23 Preparation of 2,2'-((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(4-oxopyrrolo[2,1-f][1,2,4]triazine-6,3(4H)-diyl))diacetic acid (Compound 23) [ka] Following the procedure for compound 1, the title compound 23A was obtained by replacing 1D with 2-bromomethyl acetate. Compound 23A (30 mg, 0.05 mmol, 1.0 equiv.) and LiOH (4.5 mg, 0.2 mmol, 4 equiv.) were added to methanol (2.0 mL) and HO (0.5 mL) and stirred for 1 hour. The mixture was purified by preparative HPLC to give compound 23 (1.1 mg, yield: 3.8%). LCMS (ESI): C 28 H 18 Calculated for Cl2N6O6; [M+H]+: 605.1, 607.1, Found: 605.1, 607.1. 1 H NMR (500 Mhz, DMSO-d6) 1 H NMR (500 MHz, DMSO-d6) δ 8.54 (s, 2H), 7.99 (t, J = 1.7 Hz, 4H), 7.73-7.66 (m, 4H), 7.48 (t, J = 7.6 Hz, 2H), 7.32 (dd, J = 7.5, 1.6 Hz, 2H), 7.28 (d, J = 1.7 Hz, 2H), 4.56 (d, J = 5.8 Hz, 4H)

[0215] Example 24 Preparation of 3,3'-((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(4-oxopyrrolo[2,1-f][1,2,4]triazine-6,3(4H)-diyl))dipropionic acid (Compound 24) [ka] Following the procedure for compound 23, substituting methyl 3-bromopropanoate for methyl 2-bromoacetate, gave the title compound 24. LCMS (ESI): C 30 H 22 Calculated for Cl2N6O6; [M+H]+: 633.1, 635.1, Found: 633.1, 635.1. 1 H NMR (500 Mhz, DMSO-d6) δ 8.61 (d, J = 1.6 Hz, 2H), 8.28 (s, 2H), 7.94 (d, J = 1.6 Hz, 2H), 7.68 (m, 4H), 7.61-7.31 (m, 2H), 4.22 (dq, J = 25.6, 5.9 Hz, 2H), 2.68 (t, J = 5.9 Hz, 2H).

[0216] Example 25 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(2-(2H-1,2,3-triazol-2-yl)ethyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 25) [ka] Following the procedure for compound 1, replacing 1D-1 with 2-(2H-1,2,3-triazol-2-yl)ethan-1-ol gave the title compound 25. LCMS (ESI): C 32 H 24 Cl2N 12 Calculated for O2; [M+H]+: 679.1, 681.1, Found: 679.1, 681.1. 1H NMR (500 Mhz, DMSO-d6) δ 8.54 (s, 2H), 7.99 (t, J = 1.7 Hz, 4H), 7.73-7.66 (m, 4H), 7.48 (t, J = 7.6 Hz, 2H), 7.32 (dd, J = 7.5, 1.6 Hz, 2H), 7.28 (d, J = 1.7 Hz, 2H), 4.56 (d, J = 5.8 Hz, 4H), 4.29 (t, J = 5.7 Hz, 4H).

[0217] Example 26 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(2-(4-methylpiperazin-1-yl)ethyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 26) [ka] Following the procedure for compound 1, replacing 1D-1 with 2-(4-methylpiperazin-1-yl)ethan-1-ol gave the title compound 26. LCMS (ESI): C 38 H 42 Cl2N 10 Calculated values ​​for O2: [M+H]+: 741.3, 743.3, Found: 741.3, 743.3. 1 H NMR (500 Mhz, DMSO-d6) δ 8.41 (d, J = 1.6 Hz, 2H), 8.19 (d, J = 5.1 Hz, 2H), 8.01 (d, J = 1.6 Hz, 2H), 7.68-7.65 (m, 4H), 7.52-7.49 (m, 2H), 4.06 (t, J = 6.4 Hz, 1.6 Hz, 4H), 2.64-2.55 (m, 20H), 2.29 (s, 6H).

[0218] Example 27 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-((5-oxotetrahydrofuran-2-yl)methyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 27) [ka] Following the procedure for compound 1, replacing 1D-1 with 5-(hydroxymethyl)dihydrofuran-2(3H)-one gave the title compound 27. LCMS (ESI): C 34 H 26 Calculated for Cl2N6O6; [M+H]+: 685.1, 687.1, Found: 685.1, 687.1. 1 H NMR (500 Mhz, DMSO-d6) δ 8.61 (d, J = 1.6 Hz, 2H), 8.18 (d, J = 5.1 Hz, 2H), 7.94 (d, J = 1.6 Hz, 2H), 7.68 (m, 4H), 7.52 (m, 2H), 4.95 (m, 2H), 4.43 (m, 2H), 4.36 (m, 2H), 2.40-2.13 (m, 6H), 2.13-1.93 (m, 2H).

[0219] Example 28 Preparation of 5,5'-((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(4-oxopyrrolo[2,1-f][1,2,4]triazine-6,3(4H)-diyl))bis(4-hydroxypentanoic acid) (Compound 28) [ka] Compound 27 (5.0 mg, 7.3 μmol, 1.0 equiv.) and LiOH (0.7 mg, 30 μmol, 4.0 equiv.) were dissolved in MeOH / HO (2 mL / 0.5 mL) at 25° C. and stirred for 1 hour. The reaction solution was purified by preparative liquid chromatography to give compound 28 (3.0 mg, 57% yield). LCMS (ESI): C 34 H 30Calculated for Cl2N6O8; [M+H]+: 721.1, 723.1, Found: 721.1, 723.1. 1 H NMR (500 MHz, DMSO-d6) δ 8.60 (d, J = 1.6 Hz, 2H), 8.18 (d, J = 5.1 Hz, 2H), 7.94 (d, J = 1.6 Hz, 2H), 7.68-7.65 (m, 4H), 7.52-7.50 (m, 2H), 4.96-6.92 (m, 2H), 4.43-4-42 (m, 2H), 4.36-4.33 (m, 2H), 3.90(s, 2H) 2.40-2.13 (m, 6H), 2.13-1.93 (m, 2H).

[0220] Example 29 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-2-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 29) [ka] To a stirred mixture of compound 29A (2.0 g, 9.8 mmol, 1.0 equiv.) in ACN (20 mL) was added CsCO (6.4 g, 19.6 mmol, 2.0 equiv.) and compound 29B (2.0 g, 14.7 mmol, 1.5 equiv.) at 0 °C. The resulting mixture was stirred at 25 °C for 1 h. The reaction was quenched with water and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (100 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (3:1) to give compound 29C (1.4 g, yield: 54.7%). LCMS (ESI): Calcd for C8H9BrN2O3; [M+H]+: 261.1, 263.1, Found: 261.1, 263.1.

[0221] A mixture of compound 29C (1.4 g, 5.36 mmol, 1.0 equiv) and NH3 / MeOH (7.0 M, 30 mL) was stirred in a sealed tube at 110 °C for 2 h. After concentration, the residue was purified by flash column chromatography eluting with DCM / MeOH (10:1) to give compound 29D (1.0 g, 81.9% yield). LCMS (ESI): calculated for C7H6BrN3O; [M+H]+: 228.1, 230.1, found: 228.1, 230.1.

[0222] Following the procedure for compound 1, replacing 1B with 29D gave the title compound 29. LCMS (ESI): C 36 H 32 Calculated for Cl2N8O4; [M+H]+: 711.2, 713.2, Found: 711.2, 713.2. 1 H NMR (500 MHz, DMSO-d6) δ 7.87 (d, J = 2.5 Hz, 2H), 7.81 (s, 2H), 7.78 (s, 2H), 7.56 (t, J = 10.0 Hz, 2H), 7.47 (d, J = 10.0 Hz, 2H), 7.35 (s, 2H), 4.06-3.98 (m, 4H), 3.94-3.88 (m, 2H), 2.37-2.32 (m, 2H), 2.32 (s, 6H), 2.19-2.14 (m, 2H), 2.09-2.05 (m, 2H), 1.81-1.75 (m, 2H).

[0223] Example 30 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-2-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 30) [ka] Following the procedure for compound 29, substituting methyl 4-bromo-1H-pyrrole-2-carboxylate for 29A gave the title compound 30C. LCMS (ESI): Calculated for C8H7BrN2O; [M+H]+: 226.0, 228.0, Found: 226.0, 228.0.

[0224] Following the procedure for compound 1, substituting 30C for 1B gave the title compound 30. LCMS (ESI): C 38 H 34 Calculated for Cl2N6O4; [M+H]+: 709.2, 711.2, Found: 709.2, 711.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.87 (s, 2H), 7.79 (s, 2H), 7.71-7.65 (m, 2H), 7.51-7.42 (m, 4H), 7.33-7.27 (m, 2H), 6.87 (d, J = 5.9 Hz, 2H), 4.02-3.94 (m, 3H), 3.76-3.69 (m, 3H), 2.28(s, 6H), 2.23-2.14 (m, 2H), 2.13-2.02 (m, 4H), 1.81-1.72 (m, 2H).

[0225] Example 31 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-cyclopropyl-2-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 31) [ka] Following the procedure for compound 30, substituting 2-bromo-1-cyclopropylethan-1-one for 1-bromopropan-2-one, gave the title compound 31. LCMS (ESI): C 42 H 38 Calculated for Cl2N6O4; [M+H]+: 761.2, 763.3, Found: 761.2, 763.3. 1H NMR (500 Mhz, DMSO-d6) δ 7.83 (s, 2H), 7.76 (s, 2H), 7.66-7.64 (m, 2H), 7.51-7.48 (m, 4H), 7.29-7.28 (m, 2H), 7.19 (s, 2H), 4.15-4.14 (m, 4H), 4.06 (s, 2H), 2.32-2.30 (m, 2H), 2.16-2.10 (m, 4H), 1.86-1.81 (m, 4H), 0.96-0.94 (m, 4H), 0.82-0.81 (m, 2H),0.65-0.63 (m, 2H).

[0226] Example 32 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-cyclopropyl-5-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 32) [ka] Following the procedure for compound 29, substituting 2-bromo-1-cyclopropylethan-1-one for 1-bromopropan-2-one, gave the title compound 32. LCMS (ESI): C 40 H 36 Calculated for Cl2N8O4; [M+H]+: 763.2, 765.2, Found: 763.2, 765.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.87 (d, J = 2.5 Hz,2H), 7.81 (s, 2H), 7.78 (s, 2H), 7.56 (t, J = 10.0 Hz, 2H), 7.47 (d, J = 10.0 Hz, 2H), 7.35 (s, 2H), 4.06-3.98 (m, 4H), 3.94-3.88 (m, 2H),2.35-2.33(m, 2H), 2.16-2.10 (m, 4H), 1.88-1.85 (m, 4H), 0.97-0.93 (m, 4H), 0.83-0.82 (m, 2H),0.65-0.61 (m, 2H).

[0227] Example 33 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-ethyl-5-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 33) [ka] Following the procedure for compound 29, substituting 1-bromobutan-2-one for 1-bromopropan-2-one, gave the title compound 33. LCMS (ESI): C 38 H 36 Calculated for Cl2N8O4; [M+H]+: 739.2, 741.2, Found: 739.2, 741.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.86 (d, J = 2.5 Hz, 2H), 7.82 (s, 2H), 7.78 (s, 2H), 7.57 (t, J = 10.0 Hz, 2H), 7.46 (d, J = 10.0 Hz, 2H), 7.36 (s, 2H), 4.05-3.99 (m, 4H), 3.95-3.90 (m, 2H), 2.35-2.32 (m, 2H), 2.29-2.31 (m, 4H), 2.19-2.14 (m, 2H), 2.12-2.10 (m, 6H),2.09-2.05 (m, 2H), 1.81-1.75 (m, 2H).

[0228] Example 34 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-cyclopropyl-3-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 34) [ka] To a solution of 34A (4.73 g, 23.18 mmol, 1.0 equiv) in DMF (100 mL) was added NaH (1.11 g, 60% in oil, 27.9 mmol, 1.2 equiv) at 0 °C and stirred for 10 min. Next, (aminooxy)diphenylphosphine oxide (6.5 g, 27.9 mmol, 1.2 equiv) was added to the mixture at 0 °C, and the reaction was stirred at 25 °C for 15 h. The mixture was poured into water (100 mL) and extracted with EtOAc (50 mL × 2). The layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:0 to 10:1) to give 38B (4.99 g, yield: 98.8%). LCMS (ESI): Calculated for C6H7BrN2O2; [M+H]+: 219.04, 221.03, Found: 219.35, 221.38.

[0229] To a solution of 34B (1.2 g, 5.50 mmol, 1 equiv) in cyclopropanecarbonitrile (1.53 g, 22.83 mmol, 4.15 equiv) was added HCl (4.0 M in dioxane, 3.5 mL, 16.5 mmol, 3 equiv) at 25 °C, and the mixture was stirred at 82 °C for 15 h. The reaction mixture was concentrated under reduced pressure to give 34C (1.56 g, 100% yield). LCMS (ESI): C 10 H 12 Calculated for BrN3O2; [M+H]+: 285.0, 287.0, Found: 285.0, 287.0.

[0230] To a solution of 34C (1.56 g, 5.47 mmol, 1.0 equiv) in MeCN (15 mL) was added EtN (2.0 mL) at 25 °C, and the mixture was stirred at 85 °C for 7 h. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE: EtOAc = 5:1 to 2:1) to give 34D (1.28 g, 92% yield). LCMS (ESI): calculated for CHBrNO; [M+H]: 254.09, 256.13, found: 254.00, 256.02.

[0231] Following the procedure for compound 1, replacing 1B with 34D gave the title compound 34. LCMS (ESI): C38 H 38 Calculated for Cl2N6O8; [M+H] + :830.72, Actual value:830.41. 1 H NMR (500 MHz, DMSO-d6) δ 8.23 ​​(s, 2H), 7.85 (s, 2H), 7.75-7.68 (m, 2H), 7.52 (t, J = 7.6 Hz, 2H), 7.39 (s, 2H), 7.33 (d, J = 7.4 Hz, 2H), 4.12-4.05 (m, 2H), 3.94-3.89 (m, 6H), 2.27-2.22 (m, 2H), 2.16-2.08 (m, 4H), 1.97-2.94 (M, 8H), 1.79-1.77 (m,2H).

[0232] Example 35 Preparation of 6,6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-methyl-3-(((S)-5-oxopyrrolidin-2-yl)methyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 35) [ka] Following the procedure for compound 34, substituting acetonitrile for cyclopropanecarbonitrile, gave the title compound 35. LCMS (ESI): C 36 H 32 Calculated for Cl2N8O4; [M+H] + :711.2,713.2,Actual values:711.2,713.2. 1H NMR (500 Mhz, DMSO-d6): 8.12 (s, 2H), 8.03 (d, J = 1.2 Hz, 2H), 7.85 (s, 2H), 7.72 (d, J = 6.4 Hz, 2H), 7.48 (t, J = 6.4 Hz, 2H), 7.25 (d, J = 1.6 Hz, 2H), 3.94-3.88 (m, 4H), 3.02 (s, 6H), 2.27-2.21 (m, 2H), 2.15-2.08 (m, 4H), 1.77-1.72 (m,2H).

[0233] Example 36 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-5-(((S)-pyrrolidin-2-yl)methyl)pyrazolo[1,5-a]pyrazin-4(5H)-one) dihydrochloride (Compound 36) [ka] To a stirred solution of compound 36A-1 (3.0 g, 14.9 mmol, 1.0 equiv.) in DCM (30 mL), trimethylamine (3.01 g, 29.9 mmol, 2 equiv.), DMAP (0.18 g, 1.5 mmol, 0.1 equiv.), and 4-methylbenzenesulfonyl chloride (3.70 g, 19.4 mmol, 1.30 equiv.) were added at 0° C. The resulting mixture was stirred at 25° C. for 16 h. The reaction was quenched with water (50 mL) and extracted with DCM (50 mL×3). The combined organic layer was washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give compound 36A (5.0 g, yield: 94.5%). LCMS (ESI): C 17 H 25 Calculated value for NO5S; [M+H]+: 356.1, Found: 356.1

[0234] [ka] Following the procedure for compound 1, substituting 29D for 1B and 36A for 1D gave the title compound 36C.

[0235] [ka] To a stirred mixture of compound 36C (30 mg, 0.03 mmol, 1.0 equiv) in DCM (10 mL) was added HCl / dioxane (4.0 M, 3.0 mL) at 25 °C. The resulting mixture was stirred at 25 °C for 3 h and concentrated under reduced pressure. The residue was dissolved in ACN / HO (1:4) (8.0 mL) and lyophilized to give compound 36 (24.6 mg, yield: 96.1%). LCMS (ESI): C 36 H 38 Calculated for Cl4N8O2; [M+H]+: 683.3, 685.3, Found: 683.3, 685.3. 1 H NMR (500 MHz, DMSO-d6) δ 7.90-7.84 (m, 4H), 7.60-7.54 (m, 2H), 7.49-7.44 (m, 2H), 7.40-7.36 (m, 2H), 4.45-4.35 (m, 2H), 4.29-4.21 (m, 2H), 3.74-3.64 (m, 2H), 3.52-3.43 (m, 4H), 3.13-3.02 (m, 2H), 2.39 (s, 6H), 2.19-2.09 (m, 2H), 2.06-1.93 (m, 2H), 1.92-1.86 (m, 2H), 1.80-1.70 (m, 2H).

[0236] Example 37 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-5-(((R)-pyrrolidin-2-yl)methyl)pyrazolo[1,5-a]pyrazin-4(5H)-one) dihydrochloride (Compound 37) [ka] Following the procedure for compound 36, replacing 36A-1 with tert-butyl (R)-2-(hydroxymethyl)pyrrolidine-1-carboxylate gave the title compound 37. LCMS (ESI): C 36 H 38Calculated for Cl4N8O2; [M+H]+: 683.3, 685.3, Found: 683.3, 685.3. 1 H NMR (500 MHz, DMSO-d6) δ 7.90-7.84 (m, 4H), 7.61-7.54 (m, 2H), 7.49-7.43 (m, 2H), 7.40-7.36 (m, 2H), 4.45-4.35 (m, 2H), 4.29-4.21 (m, 2H), 3.74-3.64 (m, 2H), 3.52-3.43 (m, 4H), 3.12-3.02 (m, 2H), 2.39 (s, 6H), 2.19-2.09 (m, 2H), 2.06-1.93 (m, 2H), 1.92-1.87 (m, 2H), 1.80-1.71 (m, 2H).

[0237] Example 38 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-2-(((S)-pyrrolidin-2-yl)methyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 38) [ka] Following the procedures for Compound 29 and Compound 36, substituting 4-bromo-1H-pyrrole-2-methyl carboxylate for 29A, the title compound 38 was obtained. LCMS (ESI): C 36 H 38 Calculated for Cl4N8O2; [M+H]+: 681.3, 683.3, Found: 681.3, 683.3. 1H NMR (500 MHz, DMSO-d6) δ 8.13 (s, 2H), 7.91-7.86 (m, 4H), 7.62-7.55 (m, 2H), 7.49-7.44 (m, 2H), 7.42-7.37 (m, 2H), 4.45-4.35 (m, 2H), 4.29-4.23 (m, 2H), 3.74-3.66 (m, 2H), 3.52-3.47 (m, 4H), 3.13-3.02 (m, 2H), 2.32 (s, 6H), 2.17-2.08 (m, 2H), 2.06-1.95 (m, 2H), 1.93-1.89 (m, 2H), 1.80-1.74 (m, 2H).

[0238] Example 39 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-2-(((R)-pyrrolidin-2-yl)methyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 39) [ka] Following the procedure for compound 38, replacing 36A-1 with tert-butyl (R)-2-(hydroxymethyl)pyrrolidine-1-carboxylate gave the title compound 39. LCMS (ESI): C 36 H 38 Calculated for Cl4N8O2; [M+H]+: 681.3, 683.3, Found: 681.3, 683.3. 1H NMR (500 MHz, DMSO-d6) δ 8.13 (s, 2H), 7.91-7.86 (m, 4H), 7.62-7.55 (m, 2H), 7.49-7.44 (m, 2H), 7.42-7.37 (m, 2H), 4.45-4.35 (m, 2H), 4.29-4.23 (m, 2H), 3.74-3.66 (m, 2H), 3.52-3.47 (m, 4H), 3.13-3.02 (m, 2H), 2.32 (s, 6H), 2.17-2.08 (m, 2H), 2.06-1.95 (m, 2H), 1.93-1.89 (m, 2H), 1.80-1.74 (m, 2H).

[0239] Example 40 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-2-(((R)-pyrrolidin-2-yl)methyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 40) [ka] Following the procedure for compound 29, substituting 29A with methyl 5-bromo-1H-1,2,4-triazole-3-carboxylate gave the title compound 40. LCMS (ESI): C 34 H 30 Cl2N 10 Calculated for O4; [M+H]+: 713.2, 715.2, Found: 713.2, 715.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.78 (s, 2H), 7.72-7.64 (m, 2H), 7.51-7.43 (m, 4H), 7.32-7.27 (m, 2H), 6.89 (d, J = 5.9 Hz, 2H), 4.02-3.94 (m, 3H), 3.76-3.69 (m, 3H), 2.29 (s, 6H), 2.23-2.14 (m, 2H), 2.13-2.02 (m, 4H), 1.81-1.72 (m, 2H).

[0240] Example 41 Preparation of (3S,3'S)-1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(5-methyl-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2,6-diyl))bis(methylene))bis(pyrrolidine-3-carboxylic acid) (Compound 41) [ka] Compound 29D (1.0 g, 4.4 mmol, 1 equiv.) and CsCO (2.14 g, 6.6 mmol, 1.5 equiv.) were dissolved in DMF (10 mL). CHI (0.75 g, 5.3 mmol, 1.2 equiv.) was added to the reaction mixture at 0 °C and stirred for 1 h. Water (20 mL) was added to precipitate the product, affording compound 41A (0.95 g, 89.5% yield). LCMS (ESI): calculated for C8H8BrN3O; [M+H]+: 242.0, 244.0, found: 242.0, 244.0.

[0241] Compound 1A (180 mg, 0.37 mmol, 1 equiv.), compound 41A (201 mg, 0.81 mmol, 2.2 equiv.), CsCO (618 mg, 1.85 mmol, 5.0 equiv.), and Pd(dppf)Cl·DCM (62 mg, 0.07 mmol, 0.20 equiv.) were dissolved in DMF (5.0 mL) and HO (1.0 mL) under a N atmosphere. The reaction was carried out at 90 °C for 2 h. After cooling, 30 mL of water was added, and the resulting mixture was filtered. The filter cake was washed with EtOAc (3 mL × 3) to give compound 41B (180 mg, 90.1% yield). LCMS (ESI): C 28 H 22 Calculated for Cl2N6O2; [M+H]+: 545.1, 546.1, Found: 545.1, 546.1.

[0242] Compound 41B (100 mg, 0.19 mmol, 1.0 equiv.) and selenium dioxide (404 mg, 1.89 mmol, 20 equiv.) were dissolved in dioxane (35 mL). The reaction was carried out at 90° C. for 24 hours. After cooling, 150 mL of water and 100 mL of DCM were added for extraction. The organic phase was dried over sodium sulfite and concentrated to give the residue as compound 41C (80 mg, yield: 76.2%). LCMS (ESI): C 28 H 18 Calculated for Cl2N6O4; [M+H]+: 573.1, 575.1, Found: 573.1, 575.1.

[0243] Compound 41C (40 mg, 0.07 mmol, 1 equiv.), compound 41D (80 mg, 0.7 mmol, 10 equiv.), and titanium tetraisopropanolate (187 mg, 0.7 mmol, 10 equiv.) were dissolved in THF (10 mL). The reaction was carried out at 60 °C for 2 h. After cooling to 25 °C, NaBH (26 mg, 0.7 mmol, 10 equiv.) was added, and the reaction was stirred at 25 °C. After 2 h, 2 mL of MeOH was added, the resulting mixture was filtered, and the filter cake was washed with MeOH (1 mL). The filtrate was collected and purified by preparative HPLC to give compound 41 (17.6 mg, yield: 32.6%). LCMS (ESI): C 38 H 36 Calculated for Cl2N8O6; [M+H]+: 771.1, 773.1, Found: 771.1, 773.1. 1 H NMR (500 MHz, DMSO-d6) δ 8.10 (s, 2H), 7.93-7.85 (m, 2H), 7.59-7.56 (m, 2H), 7.52-7.47 (m, 2H), 7.45 (s, 2H), 3.56-3.47 (m, 4H), 3.41 (s, 6H), 3.25-3.16 (m, 4H), 3.14-3.06 (m, 4H), 3.05-2.98 (m, 2H), 2.26-2.17 (m, 2H), 2.14-2.06 (m, 2H).

[0244] Example 42 Preparation of 2,2'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-(((2-hydroxyethyl)amino)methyl)-5-methylpyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 42) [ka] Following the procedure for compound 41, substituting 2-aminoethan-1-ol for 41D, gave the title compound 42. LCMS (ESI): C 32 H 32 Calculated for Cl2N8O4; [M+H]+: 663.2, 665.2, Found: 663.2, 665.2. 1 H NMR (500 MHz, DMSO-d6) δ 8.10 (s, 2H), 7.92-7.87 (m, 2H), 7.62-7.55 (m, 2H), 7.51-7.46 (m, 2H), 7.45 (s, 2H), 5.33 (s, 2H), 4.35 (s, 4H), 3.73-3.70 (m, 4H), 3.60-3.55 (m, 2H), 3.54 (s, 6H), 3.24-3.09 (m, 4H).

[0245] Example 43 Preparation of (3S,3'S)-1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(2-methyl-1-oxo-1,2-dihydropyrrolo[1,2-a]pyrazine-7,3-diyl))bis(methylene))bis(pyrrolidine-3-carboxylic acid) (Compound 43) [ka] Following the procedure for compound 41, substituting 30C for 29D, gave the title compound 43. LCMS (ESI): C 40 H 38 Calculated for Cl2N6O6; [M+H]+: 769.2, 771.2, Found: 769.2, 771.2. 1H NMR (500 Mhz, DMSO-d6) δ 7.90 (s, 2H), 7.69-7.68 (m, 4H), 7.49 (t, J = 6Hz, 2H), 7.37-7.32 (m, 4H), 3.72-3.52 (m, 16H), 3.44 (s, 6H).

[0246] Example 44 Preparation of N,N'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-methyl-1-oxo-1,2-dihydropyrrolo[1,2-a]pyrazine-7,3-diyl))bis(methylene))bis(azanediyl))bis(ethane-2,1-diyl))diacetamide (Compound 44) [ka] Following the procedure for compound 43, substituting N-(2-aminoethyl)acetamide for (S)-pyrrolidine-3-carboxylic acid gave the title compound 44. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O4; [M+H]+: 743.3, 745.3, Found: 743.3, 745.3. 1 H NMR (500 MHz, DMSO-d6) δ 8.59 (s, 2H), 8.15 (t, J = 5.7 Hz, 2H), 7.77 (d, J = 1.8 Hz, 2H), 7.64 (dd, J = 7.8, 1.7 Hz, 2H), 7.45 (t, J = 7.6 Hz, 2H), 7.33 (s, 2H), 7.28 (dd, J = 7.5, 1.7 Hz, 2H), 7.23 (d, J = 1.7 Hz, 2H), 4.11 (s, 6H), 3.32 -3.30 (m, 4H), 3.24 - 3.10 (m, 4H), 3.07 - 2.99 (m, 4H), 2.01 (s, 6H).

[0247] Example 45 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(((2-hydroxyethyl)amino)methyl)-2-methylpyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 45) [ka] Following the procedure for compound 43, substituting 2-aminoethan-1-ol for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 45. LCMS (ESI): C 34 H 34 Calculated for Cl2N6O4; [M+H]+: 661.2, 663.2, Found: 661.2, 663.2. 1 H NMR (500 MHz, DMSO-d6) δ 8.59 (s, 2H), 8.14 (t, J = 5.7 Hz, 2H), 7.76 (d, J = 1.8 Hz, 2H), 7.63 (dd, J = 7.8, 1.7 Hz, 2H), 7.42 (t, J = 7.6 Hz, 2H), 7.34 (s, 2H), 7.26 (dd, J = 7.5, 1.7 Hz, 2H), 4.09 (s, 6H), 3.62 (m, 2H), 3.35 -3.31 (m, 4H), 3.23 - 3.18 (m, 4H), 3.06 - 2.97 (m, 4H).

[0248] Example 46 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-(((S)-3-hydroxypyrrolidin-1-yl)methyl)-2-methylpyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 46) [ka] Following the procedure for compound 43, substituting (S)-pyrrolidin-3-ol for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 46. LCMS (ESI): C 38 H 38Calculated for Cl2N6O4; [M+H]+: 713.2, 715.2, Found: 713.2, 715.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.90 (s, 2H), 7.71-7.70 (m, 4H), 7.47 (t, J = 6 Hz, 2H), 7.32-7.26(m, 4H), 3.70-3.52 (m, 16H), 3.42 (s, 6H).

[0249] Example 47 Preparation of (3S,3'S)-1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(pyrrolidine-3-carboxylic acid) (Compound 47) [ka] To a stirred mixture of compound 29D (150 mg, 0.65 mmol, 1 equiv.) in DMF (2.0 mL) was added CsCO (536 mg, 1.64 mmol, 2.5 equiv.) and 2-bromo-1,1-dimethoxyethane (221 mg, 1.32 mmol, 2 equiv.) at 0 °C. The resulting mixture was stirred at 80 °C for 16 h. After cooling to 25 °C, the reaction was quenched with water (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (1:2) to give compound 47A (95 mg, yield: 45.7%). LCMS (ESI): C 11 H 14 Calculated for BrN3O3; [M+H]+: 316.1, 318.1, Found: 284.1, 286.1.

[0250] To a stirred mixture of compound 47A (95 mg, 0.30 mmol, 2.2 equiv) in DMF (5.00 mL) was added a solution of compound 1A (65 mg, 0.14 mmol, 1 equiv), Pd(dppf)Cl₂·DCM (22 mg, 0.03 mmol, 0.2 equiv), and Cs₂CO₃ (223 mg, 0.70 mmol, 5 equiv) in HO (1.0 mL) at 25 °C. The resulting mixture was stirred at 80 °C for 2 h. The mixture was cooled to 25 °C and quenched with water. The resulting mixture was extracted with EtOAc (30 mL × 3). The combined organic layers were washed with brine (50 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (1:2) to give compound 47B (80 mg, 84.4% yield). LCMS(ESI):C 34 H 34 Calculated for Cl2N6O6; [M+H]+: 693.2, 695.2, Found: 693.2, 695.2.

[0251] To a stirred mixture of compound 47B (65 mg, 0.09 mmol, 1 eq) in HCl / 1,4-dioxane (1.0 M, 3.00 mL), HO (3.00 mL) was added at 25° C. The resulting mixture was stirred at 85° C. for 2 hours. The mixture was cooled to 25° C. and quenched with water. The resulting mixture was extracted with DCM (30 mL×3). The combined organic layer was washed with saturated aqueous sodium bicarbonate solution (50 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give compound 47C (50 mg, yield: 89.2%). LCMS (ESI): C 30 H 22 Calculated for Cl2N6O4; [M+H]+: 601.2, 603.2, Found: 601.2, 603.2.

[0252] To a stirred mixture of compound 47C (30 mg, 0.05 mmol, 1.0 equiv) in DCM (10 mL) was added (S)-pyrrolidine-3-carboxylic acid (56 mg, 0.50 mmol, 10 equiv) and 1 drop of HOAc at 25° C. After stirring at 25° C. for 0.5 h, sodium triacetoxyborohydride (211 mg, 1 mmol, 20 equiv) was added. The resulting mixture was stirred at 25° C. for 16 h. The reaction mixture was concentrated, and the residue was dissolved in MeOH and purified by preparative liquid phase chromatography to give compound 47 (16.6 mg, yield: 41.5%). LCMS (ESI): C 40 H 40 Calculated for Cl2N8O6; [M+H]+: 799.2, 801.2, Found: 799.2, 801.2. 1H NMR (500 MHz, DMSO-d6) δ 7.87-7.81 (m, 2H), 7.78 (s, 2H), 7.73-7.68 (m, 2H), 7.59-7.52 (m, 2H), 7.47-7.43 (m, 2H), 3.92-3.72 (m, 4H), 2.93-2.83 (m, 4H), 2.81-2.64 (m, 8H), 2.64-2.56 (m, 2H), 2.30 (s, 6H), 1.97-1.87 (m, 2H), 1.83-1.74 (m, 2H).

[0253] Example 48 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-(2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)-3-methylpyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 48) [ka] Following the procedure for compound 47, substituting 30C for 29D and (S)-pyrrolidin-3-ol for (S)-pyrrolidin-3-ol gave the title compound 48. LCMS (ESI): C 40 H 42 Calculated for Cl2N6O4; [M+H]+: 741.7, 743.7, Found: 741.7, 743.7. 1 H NMR (500 MHz, DMSO-d6) δ 7.81 (s, 2H), 7.70-7.63 (m, 2H), 7.48 (t, J = 7.6 Hz, 2H), 7.36 (s, 2H), 7.29 (d, J = 7.4 Hz, 2H), 7.24 (d, J = 1.7 Hz, 2H), 5.54 (s, 2H), 4.50-4.41 (m, 2H), 4.30-4.19 (m, 4H), 3.84-3.46 (m, 8H), 3.22-3.07 (m, 4H), 2.28 (s, 6H), 2.04-1.93 (m, 2H), 1.90-1.78 (m, 2H).

[0254] Example 49 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-2-(2-((((S)-5-oxopyrrolidin-2-yl)methyl)amino)ethyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 49) [ka] Following the procedure for compound 48, substituting (S)-5-(aminomethyl)pyrrolidin-2-one for (S)-pyrrolidin-3-ol, gave the title compound 49. LCMS (ESI): C 42 H 44 Calculated for Cl2N8O4; [M+H]+: 795.3, 797.3, Found: 795.3, 797.3. 1H NMR (500 MHz, DMSO-d6) δ 8.66 (s, 2H), 7.82 (d, J = 1.8 Hz, 2H), 7.67 (d, J = 9.2 Hz, 4H), 7.48 (t, J = 7.6 Hz, 2H), 7.37 (s, 2H), 7.29 (dd, J = 7.5, 1.7 Hz, 2H), 7.24 (d, J = 1.7 Hz, 2H), 4.25-4.16 (m, 4H), 3.87-3.80 (m, 2H), 3.27-3.22 (m, 4H), 3.12-3.02 (m, 4H), 2.28 (s, 6H), 2.22-2.14 (m, 6H), 1.80-1.75 (m, 2H).

[0255] Example 50 Preparation of (2S,2'S,4S,4'S)-1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-1-oxopyrrolo[1,2-a]pyrazine-7,2(1H)-diyl))bis(ethane-2,1-diyl))bis(4-hydroxypyrrolidine-2-carboxylic acid) (Compound 50) [ka] Following the procedure for compound 48, substituting (2S,4S)-4-hydroxypyrrolidine-2-carboxylic acid for (S)-pyrrolidin-3-ol, gave the title compound 50. LCMS (ESI): C 42 H 42 Calculated for Cl2N6O8; [M+H]+: 829.7, 831.7, Found: 829.7, 831.7. 1H NMR (500 MHz, DMSO-d6) δ 7.81 (d, J = 1.8 Hz, 2H), 7.65 (dd, J = 7.8, 1.7 Hz, 2H), 7.46 (t, J = 7.6 Hz, 2H), 7.34 (s, 2H), 7.27 (dd, J = 7.7, 1.7 Hz, 2H), 7.20 (d, J = 1.7 Hz, 2H), 5.50 (s, 2H), 4.37 - 4.33 (m, 2H), 4.21 - 4.09 (m, 6H), 3.73 - 3.68 (m, 2H), 3.23 - 3.17 (m, 2H), 2.26 (s, 6H), 2.19 - 2.12 (m, 4H).

[0256] Example 51 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-(2-((2-hydroxyethyl)amino)ethyl)-3-methylpyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 51) [ka] Following the procedure for compound 48, substituting 2-aminoethan-1-ol for (S)-pyrrolidin-3-ol, gave the title compound 51. LCMS (ESI): C 36 H 38 Calculated for Cl2N6O4; [M+H]+: 689.2, 691.2, Found: 689.2, 691.2. 1H NMR (500 MHz, DMSO-d6) δ 8.61 (s, 4H), 7.81 (d, J = 1.8 Hz, 2H), 7.67 (dd, J = 7.9, 1.6 Hz, 2H), 7.48 (t, J = 7.7 Hz, 2H), 7.36 (s, 2H), 7.29 (dd, J = 7.6, 1.7 Hz, 2H), 7.23 (d, J = 1.7 Hz, 2H), 5.31 (s, 2H), 4.22 (t, J = 6.7 Hz, 4H), 3.65 (t, J = 6.7 Hz, 4H), 3.26 - 3.18 (m, 4H), 3.09 - 3.01 (m, 4H), 2.26 (s, 6H).

[0257] Example 52 Preparation of 2,2'-(2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(5-(2-((2-hydroxyethyl)amino)ethyl)-6-methylpyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 52) [ka] Following the procedure for compound 47, substituting 2-aminoethan-1-ol for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 52. LCMS (ESI): C 34 H 36 Calculated for Cl2N8O4; [M+H]+: 691.2, 693.2, Found: 691.2, 693.2. 1 H NMR (500 MHz, DMSO-d6) δ 7.86-7.83 (m, 2H), 7.78 (s, 2H), 7.72-7.69 (m, 2H), 7.58-7.53 (m, 2H), 7.46-7.44 (m, 2H), 3.85-3.71 (m, 8H), 3.31-3.25 (m, 2H), 2.94-2.86 (m, 6H), 2.80-2.74 (m, 4H), 2.30 (s, 6H).

[0258] Example 53 Preparation of (3S,3'S)-1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-1-oxopyrrolo[1,2-a]pyrazine-7,2(1H)-diyl))bis(ethane-2,1-diyl))bis(pyrrolidine-3-carboxylic acid) (Compound 53) [ka] Following the procedure for compound 48, substituting (S)-pyrrolidine-3-carboxylic acid for (S)-pyrrolidin-3-ol, gave the title compound 53. LCMS (ESI): C 42 H 42 Calculated for Cl2N6O6; [M+H]+: 797.2, 799.2, Found: 797.2, 799.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.81 (s, 2H), 7.67 (d, J = 6 Hz, 2H), 7.48 (t, J = 6 Hz, 2H), 7.35 (s, 2H), 7.28 (d, J = 6 Hz, 2H), 7.23 (s, 2H), 3.92-3.82 (m, 4H), 2.93-2.85 (m, 4H), 2.80-2.65 (m, 8H), 2.64-2.55 (m, 2H), 2.29 (s, 6H), 1.97-1.89 (m, 2H), 1.84-1.76 (m, 2H).

[0259] Example 54 Preparation of N,N'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-1-oxopyrrolo[1,2-a]pyrazine-7,2(1H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(ethane-2,1-diyl))diacetamide (Compound 54) [ka] Following the procedure for compound 48 and substituting N-(2-aminoethyl)acetamide for (S)-pyrrolidin-3-ol gave the title compound 54. LCMS (ESI): C40 H 44 Calculated for Cl2N8O4; [M+H]+: 771.3, 773.3, Found: 771.3, 773.3. 1 H NMR (500 MHz, DMSO-d6) δ 8.61 (s, 2H), 8.13 (t, J = 5.7 Hz, 2H), 7.81 (d, J = 1.8 Hz, 2H), 7.67 (dd, J = 7.8, 1.7 Hz, 2H), 7.48 (t, J = 7.6 Hz, 2H), 7.36 (s, 2H), 7.29 (dd, J = 7.5, 1.7 Hz, 2H), 7.24 (d, J = 1.7 Hz, 2H), 4.25-4.13 (m, 4H), 3.32 (q, J = 6.1 Hz, 4H), 3.27-3.19 (m, 4H), 3.07-2.99 (m, 4H), 2.27 (s, 6H), 1.85 (s, 6H).

[0260] Example 55 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-2-(2-((pyrrolidin-2-ylmethyl)amino)ethyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 55) [ka] Following the procedure for compound 48 and substituting tert-butyl 2-(aminomethyl)pyrrolidine-1-carboxylate for (S)-pyrrolidin-3-ol gave the title compound 55A.

[0261] To a stirred solution of 55A (20 mg, 0.021 mmol, 1 equiv) in MeOH (1.0 mL) was added HCl / dioxane (4.0 M, 1.0 mL) and stirred at 25° C. for 4 h. The reaction mixture was concentrated and purified by preparative HPLC to give compound 55 (7.9 mg, 51% yield). LCMS (ESI): C 42 H 48 Calculated for Cl2N8O2.4HCl; [M+H]+: 767.6, 769.6, Found: 767.6, 769.6. 1 H NMR (500 MHz, DMSO-d6) δ 9.28 (s, 4H), 9.10 (s, 4H),7.82 (s, 2H), 7.68-7.66 (m, 2H), 7.47 (t, J = 7.5 Hz, 2H), 7.37 (s, 2H), 7.29 (d, J = 5.0 Hz, 2H), 7.24 (d, J = 5.0 Hz, 2H), 4.24-4.18 (m, 4H), 3.83 - 3.75 (m, 4H), 3.33 - 3.25 (m, 8H), 2.65 - 2.60 (m, 2H), 2.29 (s, 6H), 2.16 - 2.08 (m, 2H), 2.00 - 1.93 (m, 2H), 1.90 - 1.83 (m, 2H), 1.73 - 1.65 (m, 2H).

[0262] Example 56 Preparation of 4,4'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-1-oxopyrrolo[1,2-a]pyrazine-7,2(1H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxybutanoic acid) (Compound 56) [ka] Following the procedure for compound 48, substituting 4-amino-3-hydroxybutanoic acid for (S)-pyrrolidin-3-ol, gave the title compound 56. LCMS (ESI): C 40 H 42 Calculated for Cl2N6O8; [M+H] + :805.2,807.2,Actual values:805.2,807.2. 1H NMR (500 MHz, DMSO-d6) δ 8.43 (s, 2H), 7.81 (dd, J = 4.9, 1.7 Hz, 2H), 7.67 (dd, J = 7.8, 1.8 Hz, 2H), 7.48 (t, J = 7.7 Hz, 2H), 7.36 (d, J = 6.8 Hz, 2H), 7.29 (d, J = 7.6 Hz, 2H), 7.23 (dd, J = 4.9, 1.6 Hz, 2H), 5.66 (s, 2H), 3.62 (t, J = 4.2 Hz, 2H), 3.27-3.16 (m, 4H), 3.09 (d, J = 12.0 Hz, 4H), 2.99-2.88 (m, 4H), 2.43 (dd, J = 23.3, 6.3 Hz, 4H), 2.26 (s, 6H).

[0263] Example 57 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 57) [ka] Following the procedure for compound 47, substituting L-serine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 57. LCMS (ESI): C 36 H 36 Calculated for Cl2N8O8; [M+H] + :779.2,781.2,Actual values:779.2,781.2. 1H NMR (500 MHz, DMSO-d6): δ 7.87-7.81 (m, 2H), 7.78 (s, 2H), 7.73-7.68 (m, 2H), 7.59-7.52 (m, 2H), 7.47-7.43 (m, 2H), 4.99-4.92 (m, 2H), 3.95-3.80 (m, 6H), 3.70-3.58 (m, 4H), 3.50-3.41 (m, 2H), 2.99-2.87 (m, 4H), 2.30 (s, 6H).

[0264] Example 58 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-1-oxopyrrolo[1,2-a]pyrazine-7,2(1H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(4-hydroxybutanoic acid) (Compound 58) [ka] Following the procedure for compound 48, substituting L-homoserine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 58. LCMS (ESI): C 40 H 42 Calculated for Cl2N6O8; [M+H] + :805.2,807.2,Actual values:805.2,807.2. 1 H NMR (500 MHz, DMSO-d6) δ 8.35 (s, 2H), 7.80 (s, 2H), 7.73-7.65 (m, 2H), 7.47 (t, J = 7.6 Hz, 2H), 7.35 (s, 2H), 7.28 (d, J = 7.4 Hz, 2H), 7.23 (d, J = 1.7 Hz, 2H), 5.56 (s, 2H), 4.54-4.44 (m, 4H), 4.34-4.23 (m, 4H), 4.08-3.94 (m, 4H), 3.55-3.30 (m, 2H), 2.28 (s, 6H), 1.88-1.76 (m, 2H).

[0265] Example 59 Preparation of (2S,2'S,4R,4'R)-1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-1-oxopyrrolo[1,2-a]pyrazine-7,2(1H)-diyl))bis(ethane-2,1-diyl))bis(4-hydroxypyrrolidine-2-carboxylic acid) (Compound 59) [ka] Following the procedure for compound 48, substituting (2R,4S)-4-hydroxypyrrolidine-2-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 59. LCMS (ESI): C 42 H 42 Calculated for Cl2N6O8; [M+H]+: 829.2, 831.2, Found: 829.2, 831.2. 1H NMR (500 MHz, DMSO-d6) δ 8.35 (s, 2H), 7.80 (s, 2H), 7.73-7.65 (m, 2H), 7.47 (t, J = 7.6 Hz, 2H), 7.35 (s, 2H), 7.28 (d, J = 7.4 Hz, 2H), 7.23 (d, J = 1.7 Hz, 2H), 5.56 (s, 2H), 4.54-4.44 (m, 4H), 4.34-4.23 (m, 4H), 4.08-3.94 (m, 4H), 3.55-3.30 (m, 2H), 2.28 (s, 6H), 1.88-1.76 (m, 2H).

[0266] Example 60 Preparation of (2S,2'S,3R,3'R)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxybutanoic acid) (Compound 60) [ka] Following the procedure for compound 47, substituting L-threonine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 60. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O8; [M+H] + :807.2,809.2,Actual values:807.2,809.2. 1 H NMR (500 MHz, DMSO-d6): δ 7.91-7.86 (m, 2H), 7.79 (s, 2H), 7.59-7.52 (m, 2H), 7.48-7.42 (m, 2H), 7.35 (s, 2H), 4.14-4.03 (m, 4H), 3.82-3.72 (m, 4H), 3.07-2.97 (m, 4H), 2.82-2.74 (m, 4H), 2.38 (s, 6H), 1.23 (s, 6H).

[0267] Example 61 Preparation of (2R,2'R)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 61) [ka] Following the procedure for compound 47, substituting D-serine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 61. LCMS (ESI): C 36 H 36 Calculated for Cl2N8O8; [M+H] + :779.2,781.2,Actual values:779.2,781.2. 1H NMR (500 MHz, DMSO-d6): δ 7.87-7.81 (m, 2H), 7.78 (s, 2H), 7.73-7.68 (m, 2H), 7.59-7.52 (m, 2H), 7.47-7.43 (m, 2H), 4.99-4.92 (m, 2H), 3.95-3.80 (m, 6H), 3.70-3.58 (m, 4H), 3.50-3.41 (m, 2H), 2.99-2.87 (m, 4H), 2.30 (s, 6H).

[0268] Example 62 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(4-hydroxybutanoic acid) (Compound 62) [ka] Following the procedure for compound 47, substituting L-homoserine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 62. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O8; [M+H] + :807.2,809.2,Actual values:807.2,809.2. 1 H NMR (500 MHz, DMSO-d6): δ 8.36 (s, 2H), 7.88 (d, J = 7.7 Hz, 2H), 7.78 (s, 2H), 7.58-7.51 (m, 2H), 7.44 (d, J = 7.5 Hz, 2H), 7.35 (s, 2H), 5.38-5.25 (m, 2H), 4.12-4.06 (m, 4H), 3.50-3.46 (m, 6H), 3.17-3.13 (m, 2H), 2.99-2.93 (m, 2H), 2.85-2.77 (m, 2H), 2.37 (s, 6H), 2.04-1.94 (m, 4H).

[0269] Example 63 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-1-oxopyrrolo[1,2-a]pyrazine-7,2(1H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 63) [ka] Following the procedure for compound 48, substituting L-serine for (S)-pyrrolidin-3-ol, gave the title compound 63. LCMS (ESI): C 38 H 38 Calculated for Cl2N6O8; [M+H] + :777.2,779.2,Actual values:777.2,779.2. 1 H NMR (500 MHz, DMSO-d6) δ 8.23 ​​(s, 2H), 7.82 (s, 2H), 7.72-7.64 (m, 2H), 7.49 (t, J = 7.6 Hz, 2H), 7.37 (s, 2H), 7.30 (d, J = 7.4 Hz, 2H), 7.25 (d, J = 1.7 Hz, 2H), 5.58 (s, 2H), 4.50-4.41 (m, 4H), 4.30-4.19 (m, 4H), 4.01-3.87 (m, 4H), 3.58-3.33 (m, 2H), 2.29 (s, 6H).

[0270] Example 64 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-methyl-4-oxopyrrolo[2,1-f][1,2,4]triazine-6,3(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 64) [ka] Following the procedure for compound 34, replacing cyclopropanecarbonitrile with acetonitrile, gave the core structure 6-bromo-2-methylpyrrolo[2,1-f][1,2,4]triazin-4(3H)-one. Following the procedure for compound 47, replacing 29D with 6-bromo-2-methylpyrrolo[2,1-f][1,2,4]triazin-4(3H)-one and (S)-pyrrolidine-3-carboxylic acid with L-serine gave the title compound 64. LCMS (ESI): C 36 H 36 Calculated for Cl2N8O8; [M+H] + :779.2,781.2,Actual values:779.2,781.2. 1 H NMR (500 MHz, DMSO-d6) δ 7.88 (s, 2H), 7.83 (s, 2H), 7.54-7.53 (m, 2H), 7.45-7.43 (m, 2H), 7.36 (s, 2H), 4.02 (s, 4H), 3.78 (s, 2H), 3.70 (s, 2H), 2.99-2.96 (m, 4H), 2.37 (s, 6H), 2.03-2.01 (m, 5H), 1.23 (s, 4H).

[0271] Example 65 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-ethyl-4-oxopyrrolo[2,1-f][1,2,4]triazine-6,3(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 65) [ka] Compound 34B (1.0 g, 4.6 mmol, 1.0 equivalent) was dissolved in HOAc (10 mL) at 0° C., and propionyl chloride (634 mg, 6.85 mmol, 1.5 equivalent) was added to the mixture and stirred for 4 hours. The mixture was concentrated and purified by silica gel column chromatography eluting with PE / EtOAc (3:1) to give compound 65A (0.80 g, yield: 63.7%). LCMS (ESI): CH11 Calculated for BrN2O3; [M+H] + :275.0,277.0,Actual values:275.0,277.0.

[0272] A mixture of compound 65A (0.80 g, 2.91 mmol) and NH3 / CH3OH (7.0 M, 20 mL) was stirred in a sealed tube at 110 °C for 2 h. After concentration, the residue was purified by flash column chromatography eluting with DCM / MeOH (10:1) to give compound 65B (0.43 g, 61.1% yield). LCMS (ESI): calculated for C8H8BrN3O; [M+H]+: 251.0, 253.0, found: 251.0, 253.0.

[0273] Following the procedure for compound 47, substituting 65B for 29D and L-serine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 65. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O8; [M+H] + :807.2,809.2,Actual values:807.2,809.2. 1 H NMR (500 MHz, DMSO-d6) δ 7.89 (s, 2H), 7.83 (s, 2H), 7.53-7.52 (m, 2H), 7.46-7.44 (m, 2H), 7.37 (s, 2H), 4.02 (s, 4H), 3.78 (s, 2H), 3.70 (s, 2H), 2.99-2.94 (m, 4H), 2.34-2.33 (m, 4H), 2.05-2.03 (m, 5H), 1.96-1.94 (m, 6H), 1.23 (s, 4H).

[0274] Example 66 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-ethyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(methylene))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 66) [ka] Following the procedure for compound 29, the core structure 2-bromo-6-ethylpyrazolo[1,5-a]pyrazin-4(5H)-one was obtained by replacing 1-bromopropan-2-one with 1-bromobutan-2-one.

[0275] Following the procedure for compound 47, substituting 2-bromo-6-ethylpyrazolo[1,5-a]pyrazin-4(5H)-one for 29D and L-serine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 66. LCMS (ESI): C 36 H 36 Calculated for Cl2N8O8; [M+H]+: 779.2, 781.2, Found: 779.2, 781.2. 1 H NMR (500 MHz, DMSO-d6) δ 7.85-7.81 (m, 2H), 7.77 (s, 2H), 7.72-7.69 (m, 2H), 7.58-7.53 (m, 2H), 7.47-7.44 (m, 2H), 4.99-4.93 (m, 2H), 3.91-3.82 (m, 4H), 3.71-3.59 (m, 4H), 3.51-3.42 (m, 2H), 2.99-2.87 (m, 4H), 2.37-2.35 (m, 4H), 1.97-1.91 (m, 6H).

[0276] Example 67 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-aminopropanoic acid) (Compound 67) [ka] Following the procedure for compound 55 and substituting (S)-2-amino-3-((tert-butoxycarbonyl)amino)propanoic acid for tert-butyl 2-(aminomethyl)pyrrolidine-1-carboxylate gave the title compound 67. LCMS (ESI): C 36 H 38 Cl2N 10 Calculated for O6; [M+H] + :777.2,779.2,Actual value:777.2,779.2.581. 1 H NMR (500 MHz, DMSO-d6): δ 7.86-7.83 (m, 2H), 7.79 (s, 2H), 7.71-7.69 (m, 2H), 7.57-7.53 (m, 2H), 7.46-7.44 (m, 2H), 4.32-4.28 (m, 2H), 3.93-3.80 (m, 4H), 3.48-3.45 (m, 2H), 3.15(t, J = 10.0 Hz, 4H) 3.00-2.90 (m, 8H), 2.31 (s, 6H).

[0277] Example 68 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-acetamidopropanoic acid) (Compound 68) [ka] Compound 68A (1.0 g, 4.2 mmol, 1 eq) and trimethylamine (850 mg, 8.4 mmol, 2 eq) were dissolved in DCM (10 mL) at 0° C. Acetyl chloride (395 mg, 5.04 mmol, 1.2 eq) was added dropwise to the reaction mixture and stirred for 1 h. Water (10 mL) was added for extraction. The organic layer was concentrated to give compound 68B (0.82 g, yield: 69.7%). LCMS (ESI): C 13 H 16 Calculated for N2O5; [M+H] + :281.1, Actual value:281.1.

[0278] Compound 68B (0.50 g, 1.78 mmol, 1 eq) and Pd / C (10% wet, 50 mg) were added to CHOH (10 mL) under H atmosphere at 25 °C and stirred for 5 h. The reaction mixture was filtered and concentrated to give compound 68C (250 mg, yield: 95.9%). LCMS (ESI): CH 10 Calculated for N2O3; [M+H] + :147.1, Actual value:147.1.

[0279] Following the procedure for compound 47, substituting compound 68C for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 68. LCMS (ESI): C 40 H 42 Cl2N 10 Calculated for O8; [M+H] + :861.2,863.2,Actual values:861.2,863.2. 1 H NMR (500 MHz, DMSO-d6): δ 7.85-7.82 (m, 2H), 7.78 (s, 2H), 7.72-7.69 (m, 2H), 7.58-7.54 (m, 2H), 7.50-7.47 (t, J = 7.5 Hz, 2H), 7.46-7.44 (m, 2H), 4.64-4.60 (m, 2H), 3.93-3.78 (m, 6H), 3.36-3.32 (m, 4H), 2.94-2.90 (m, 4H), 2.30 (s, 6H), 1.95 (s, 6H).

[0280] Example 69 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-(dimethylamino)propanoic acid) (Compound 69) [ka] Compound 68A (1.0 g, 4.2 mmol, 1.0 equiv.) and HCHO (37% in water, 4.0 mL) were added to CHOH (10 mL) at 25° C. and stirred for 30 minutes. Sodium triacetoxyborohydride (4.45 g, 21 mmol, 5 equiv.) was added to the reaction mixture and stirred for 2 hours. The reaction mixture was concentrated and purified by preparative HPLC to give compound 69A (0.90 g, yield: 80.5%). LCMS (ESI): C 13 H 18 Calculated for N2O4; [M+H] + :266.1,Actual value:266.1.

[0281] Compound 69A (0.90 g, 3.4 mmol, 1 eq) and Pd / C (10% wet, 90 mg) were added to CHOH (10 mL) under H atmosphere at 25 °C and stirred for 5 h. The reaction mixture was filtered and concentrated to give compound 69B (430 mg, yield: 96.3%). LCMS (ESI): CH 12 Calculated for N2O2; [M+H] + :132.1, Actual value:132.1.

[0282] Following the procedure for compound 47, substituting compound 69B for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 69. LCMS (ESI): C 40 H 46 Cl2N 10 Calculated for O6; [M+H] + :833.3,835.3,Actual values:833.3,835.3. 1 H NMR (500 MHz, DMSO-d6): δ 7.86-7.82 (m, 2H), 7.79 (s, 2H), 7.73-7.69 (m, 2H), 7.58-7.55 (m, 2H), 7.50-7.47 (t, J = 7.5 Hz, 2H), 7.46-7.43 (m, 2H), 4.64-4.61 (m, 2H), 3.94-3.78 (m, 6H), 3.36-3.33 (m, 4H), 2.94-2.90 (m, 4H), 2.37 (s, 12H), 2.30 (s, 6H).

[0283] Example 70 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(propane-3,1-diyl))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 70) [ka] Following the procedure for compound 47, substituting 3-bromo-1,1-dimethoxypropane for 2-bromo-1,1-dimethoxyethane and L-serine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 70. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O8; [M+H] + :807.2,809.2,Actual values:807.2,809.2. 1 H NMR (500 MHz, DMSO-d6): 1 H NMR (500 Mhz, DMSO-d6) δ 7.88 (s, 2H), 7.83 (s, 2H), 7.54-7.53 (m, 2H), 7.44-7.43 (m, 2H), 7.36 (s, 2H), 4.02 (s, 4H), 3.78 (s, 2H), 3.70 (s, 2H), 2.99-2.97 (m, 4H), 2.37 (s, 6H), 2.02-2.00 (m, 5H), 1.23 (s, 4H).

[0284] Example 71 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(5-methyl-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2,6-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 71) [ka] To a mixture of (methoxymethyl)triphenylphosphonium chloride (215 mg, 0.63 mmol, 1.2 equiv.) and dry THF (10 mL), LiHMDS (1.6 M, 0.39 mL, 1.5 equiv.) was added and stirred at 0° C. for 30 minutes. Compound 41C (300 mg, 0.52 mmol, 1.0 equiv.) was added and stirred at 25° C. for 1 hour. The reaction was quenched with water (10 mL) and extracted with EtOAc (10 mL×3). The combined organic layers were washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated and purified by silica gel column chromatography eluting with PE / EtOAc (1:2) to give compound 71A (180 mg, yield: 54.7%). LCMS (ESI): C 32 H 26 Calculated for Cl2N6O4; [M+H] + :628.1,630.1,Actual value:628.1,630.1.

[0285] A mixture of compound 71A (0.18 g, 0.397 mmol) and HCl / MeOH (1.0 M, 20 mL) was stirred at 25° C. for 1 hour, and the mixture was concentrated under reduced pressure to give compound 71B (130 mg, yield: 54.7%). LCMS (ESI): C 30 H 22 Calculated for Cl2N6O4; [M+H] + :601.1,603.1,Actual value:601.1,603.1.

[0286] Following the procedure for compound 47, compound 47C was replaced with compound 71B and (S)-pyrrolidine-3-carboxylic acid was replaced with L-serine to give the title compound 71. LCMS (ESI): C 36 H 36 Calculated for Cl2N8O8; [M+H] + :779.2,781.2,Actual values:779.2,781.2. 1H NMR (500 Mhz, DMSO-d6) δ 7.84-7.82 (m, 2H), 7.70-7.69 (m, 2H), 7.66 (s, 2H), 7.62 (t, J = 1.0 Hz, 2H), 7.56-7.50 (m, 2H), 4.97-4.79 (m, 2H), 3.92 (t, J = 5.6 Hz, 2H), 3.78-3.54 (m, 4H), 3.53-3.45 (m, 2H), 3.39 (s, 6H), 2.98 -2.85(m, 4H), 2.85-2.66 (m, 4H).

[0287] Example 72 Preparation of N,N'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(1-oxo-1,2-dihydropyrrolo[1,2-a]pyrazine-7,3-diyl))bis(methylene))bis(azanediyl))bis(ethane-2,1-diyl))diacetamide (Compound 72) [ka] Following the procedure for compound 41, substituting 7-bromo-3-methylpyrrolo[1,2-a]pyrazin-1(2H)-one for 41A and N-(2-aminoethyl)acetamide for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 72. LCMS (ESI): C 36 H 36 Calculated for Cl2N8O4; [M+H] + :715.2,717.2,Actual values:715.2,717.2. 1H NMR (500 Mhz, DMSO-d6) δ: 10.52 (s, 2H), 8.14(t, J = 4.8, 2H), 7.99 (s, 2H), 7.72-7.70 (m, 2H), 7.55 (s, 2H), 7.49 (t, J = 6.4Hz, 2H), 7.33-7.30 (m, 4H), 4.02 (s, 4H), 3.74 (s, 1H), 3.36-3.32 (m, 4H), 3.04 (s, 4H), 1.85 (s, 6H).

[0288] Example 73 Preparation of N,N'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(1-oxo-2-(((S)-5-oxopyrrolidin-2-yl)methyl)-1,2-dihydropyrrolo[1,2-a]pyrazine-7,3-diyl))bis(methylene))bis(azanediyl))bis(ethane-2,1-diyl))diacetamide (Compound 73) [ka] Compound 72 (200 mg, 0.28 mmol, 1.0 equiv.), EtN (142 mg, 1.4 mmol, 5 equiv.), BocO (306 mg, 1.4 mmol, 5 equiv.), and 4-dimethylaminopyridine (3.4 mg, 0.028 mmol, 0.1 equiv.) were added to DCM (5 mL) and stirred for 2 h. Water (10 mL) was added to the reaction mixture, which was extracted with DCM (10 mL x 3). The organic layer was then concentrated to give compound 73A (280 mg), which was used in the next step without purification. LCMS (ESI): C 46 H 52 Calculated for Cl2N8O8; [M+H] + :915.3,917.3,Actual values:915.3,917.3.

[0289] Compound 73A (280 mg, 0.31 mmol, 1.0 equiv), CsCO (398 mg, 1.22 mmol, 4 equiv), and compound 1D (247 mg, 0.92 mmol, 3.0 mmol) were added to DMF (3.0 mL) and stirred at 65 °C for 1 h. The reaction was filtered, and the filtrate was purified by preparative HPLC to give compound 73B (25 mg, 21.5%). LCMS (ESI): C 56 H 66 Cl2N 10 O 10 Calculated value for [M+H] + :1109.4,1111.4,Actual values:1109.4,1111.4.

[0290] Compound 73B (25 mg, 22.5 μmol) was added to trifluoroacetic acid (3 mL) and stirred at 25° C. for 1 hour. The reaction was purified by preparative HPLC to give compound 73 (10 mg, yield: 48.8%). LCMS (ESI): C 46 H 50 Cl2N 10 Calculated for O6; [M+H] + :909.3,911.3,Actual values:909.3,911.3. 1 H NMR (500 MHz, DMSO-d6) δ 8.59 (s, 2H), 8.15 (t, J = 5.7 Hz, 2H), 7.77 (d, J = 1.8 Hz, 2H), 7.64 (dd, J = 7.8, 1.7 Hz, 2H), 7.45 (t, J = 7.6 Hz, 2H), 7.33 (s, 2H), 7.28 (dd, J = 7.5, 1.7 Hz, 2H), 7.23 (d, J = 1.7 Hz, 2H), 4.02-3.94 (m, 3H), 3.76-3.69 (m, 3H), 3.32 -3.30 (m, 4H), 3.24 - 3.1 (m, 4H), 3.15-3.13(m, 2H), 3.07 - 2.99 (m, 4H), 2.23-2.14 (m, 2H), 2.13-2.04 (m, 4H), 2.01 (s, 6H), 1.81-1.72 (m, 2H).

[0291] Example 74 Preparation of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-methyl-3-(((((S)-5-oxopyrrolidin-2-yl)methyl)amino)methyl)pyrrolo[1,2-a]pyrazin-1(2H)-one) (Compound 74) [ka] Following the procedure for compound 43, substituting (S)-5-(aminomethyl)pyrrolidin-2-one for (S)-pyrrolidine-3-carboxylic acid gave the title compound 74. LCMS (ESI): C 40 H 40Calculated for Cl2N8O4; [M+H] + :767.2,769.2,Actual values:767.2,769.2. 1 H NMR (500 Mhz, DMSO-d6) δ: 7.95 (s, 2H), 7.73-7.71 (m, 4H), 7.60 (s, 2H), 7.50 (t, J = 6Hz, 2H), 7.31-7.28 (m, 4H), 4.27 (s, 4H), 3.88 (s, 4H), 3.47 (s, 6H), 3.17 (s, 2H), 2.23-2.14 (m, 8H), 1.82-1.80 (m, 2H).

[0292] Example 75 Preparation of 6'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2-(((S)-pyrrolidin-2-yl)methyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 75) [ka] Compound 75A (534 mg, 2.28 mmol, 1.02 equiv.), DIPEA (500 mg, 4.56 mmol, 2.0 equiv.), and HATU (1.12 g, 2.74 mmol, 1.2 equiv.) were added to DMF (5 mL) and stirred at 25° C. for 15 minutes. Compound 34B (500 mg, 2.28 mmol, 1.0 equiv.) was added to the reaction mixture and stirred at 25° C. overnight. Water (20 mL) was added to the reaction mixture, which was then extracted with DCM (20 mL×3). The combined organics were dried over Na2SO4, filtered, concentrated, and purified by silica gel column chromatography eluting with PE:EtOAc (1:1) to give 75B (355 mg, yield: 35.4%). LCMS (ESI): C 17 H 24 Calculated for BrN3O5; [M+H]+: 430.3, 432.3, Found: 430.3, 432.3.

[0293] A mixture of compound 75B (335 mg, 0.778 mmol, 1 equiv) and NH3 / MeOH (7.0 M, 15 mL) was stirred in a sealed tube at 110 °C for 8 h. After concentration, the residue was purified by flash column chromatography eluting with PE / EtOAc (1:1) to give compound 75C (278 mg, yield: 86.9%). LCMS (ESI): C 16 H 21 Calculated for BrN4O3; [M+H]+: 397.2, 399.2, Found: 397.2, 399.2, 297.2, 299.2.

[0294] Compound 1A (54 mg, 0.114 mmol, 1 equiv.), compound 75C (100 mg, 0.25 mmol, 2.2 equiv.), CsCO (200 mg, 1.25 mmol, 5 equiv.), and Pd(dppf)Cl·DCM (30 mg, 0.07 mmol, 0.25 equiv.) were dissolved in DMF (5.0 mL) and HO (1.0 mL) under N. The reaction was carried out at 90 °C for 3 h. After cooling, 30 mL of water was added, and the resulting mixture was filtered. The filter cake was washed with ethyl acetate (3 mL × 3) to give compound 75D (21 mg, 22.1% yield). LCMS (ESI): C 44 H 48 Calculated for Cl2N8O8; [M+H]+: 855.8, 857.8, Found: 855.8, 857.8.

[0295] To a solution of 75D (54 mg, 0.025 mmol, 1 equiv) was added a solution of MeOH and HCl in dioxane (4.0 M, 2.0 mL) at 25° C. The mixture was stirred at 25° C. for 4 h. The reaction mixture was concentrated and purified by preparative HPLC to give compound 75 (8.3 mg, yield: 50.2%). LCMS (ESI): C 34 H 32 Calculated for Cl2N8O2.2HCl; [M+H]+: 655.5, 657.5, Found: 655.5, 657.5. 1H NMR (500 MHz, DMSO-d6): δ 11.95 (s, 2H), 9.38 (s, 2H), 9.18 (s, 2H), 7.96 (s, 2H), 7.71 (d, J = 2.5 Hz, 2H), 7.46 (d, J = 5.0 Hz, 2H), 7.31 (d, J = 2.5 Hz, 2H), 7.24 (s, 2H), 3.90 (t, J = 5.0 Hz, 2H), 3.34 - 3.30 (m, 4H), 3.19 - 3.17 (m, 2H), 3.08 - 3.03 (m, 2H), 2.24 - 2.17 (m, 2H), 2.00 - 1.97 (m, 2H), 1.91 - 1.86 (m, 2H), 1.75 - 1.66 (m, 2H).

[0296] Example 76 Preparation of 6,6'-(2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-2-(((S)-pyrrolidin-2-yl)methyl)pyrrolo[2,1-f][1,2,4]triazin-4(3H)-one) (Compound 76) [ka] To a solution of compound 75D (50 mg, 0.05 mmol, 1.0 equiv) in DMF (2.5 mL) was added CsCO (85 mg, 0.25 mmol, 5.0 equiv) and stirred at 25 °C for 10 min, followed by the addition of CHCl (48 mg, 0.30 mmol, 6.0 equiv). The reaction mixture was stirred for 1 h. The reaction mixture was added to water (20 mL), and the solution was extracted with DCM (20 mL × 3). The combined organics were dried over NaSO, filtered, concentrated, and purified by silica gel column chromatography eluting with PE / EtOAc (1:1) to give compound 76A (30 mg, yield: 58.4%). LCMS (ESI): C 46 H 52 Calculated for Cl2N8O6; [M+H]+: 883.8, 885.8, Found: 883.8, 885.8.

[0297] To a solution of compound 76A (30 mg, 0.034 mmol, 1.0 equiv) was added MeOH (2 mL) and a solution of HCl in dioxane (4.0 M, 2.0 mL) at 25° C. The mixture was stirred at 25° C. for 4 h. The reaction mixture was concentrated and purified by preparative HPLC to give compound 76 (10 mg, yield: 43.1%). LCMS (ESI): C 36 H 36 Calculated for Cl2N8O2; [M+H]+: 682.2, 684.2, Found: 682.2, 684.2. 1 H NMR (500 MHz, DMSO-d6) δ 9.35 (s, 2H), 9.14 (s, 2H), 8.00 (d, J = 5.0 Hz, 2H), 7.73 (d, J = 2.5 Hz, 2H), 7.50 (d, J = 7.5 Hz, 2H), 7.33 (d, J = 10.0 Hz, 2H), 7.26 (s, 2H), 4.03 -3.99 (m, 2H), 3.43 (s, 6H), 3.34 - 3.30 (m, 4H), 3.23 - 3.17 (m, 4H), 2.27 - 2.20 (m, 2H), 2.03 - 1.97 (m, 2H), 1.94 - 1.86 (m, 2H), 1.79 - 1.71 (m, 2H).

[0298] Example 77 Preparation of diethyl 2,2'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))(2S,2'S)-bis(3-hydroxypropanoate) (Compound 77) [ka] Following the procedure for compound 47, substituting L-ethyl serine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 77. LCMS (ESI): C 40 H 44Calculated for Cl2N8O8; [M+H]+: 835.2, 837.2, Found: 835.2, 837.2. 1 H NMR (500 MHz, DMSO-d6) δ 7.90-7.83 (m, 4H), 7.60-7.53 (m, 2H), 7.48-7.43 (m, 2H), 7.38 (s, 2H), 4.40-4.19 (m, 8H), 3.95-3.85 (m, 3H), 3.58-3.33 (m, 11H), 2.36 (s, 6H), 1.23-1.25 (m, 6H).

[0299] Example 78 Preparation of diethyl 1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))(3S,3'S)-bis(pyrrolidine-3-carboxylic acid) (Compound 78) [ka] Following the procedure for compound 47, substituting (S)-ethyl pyrrolidine-3-carboxylate for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 78. LCMS (ESI): C 44 H 48 Calculated for Cl2N8O6; [M+H]+: 855.3, 857.3, Found: 855.3, 857.3. 1H NMR (500 MHz, DMSO-d6) δ 7.86-7.82 (m, 2H), 7.78 (s, 2H), 7.72-7.69 (m, 2H), 7.56 (dd, J = 8.6, 7.7 Hz, 2H), 7.46-7.44 (m, 2H), 4.21-4.08 (m, 4H), 3.93-3.73 (m, 4H), 2.94-2.85 (m, 4H), 2.82-2.69 (m, 6H), 2.69-2.64 (m, 2H), 2.63-2.57 (m, 2H), 2.30 (s, 6H), 2.00-1.90 (m, 2H), 1.85-1.75 (m, 2H), 1.28-1.21 (m, 6H).

[0300] Example 79 Preparation of 1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(piperidine-2-carboxylic acid) (Compound 79) [ka] Following the procedure for compound 47, substituting piperidine-2-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 79. LCMS (ESI): C 42 H 44 Calculated for Cl2N8O6; [M+H]+: 827.3, 829.3, Found: 827.3, 829.3. 1H NMR (500 MHz, DMSO-d6) δ 7.88 - 7.85 (m, 2H), 7.84 (s, 2H), 7.74 - 7.71 (m, 2H), 7.59 - 7.55 (m, 2H), 7.47 - 7.46 (m, 2H), 3.97 - 3.91 (m, 2H), 3.87 - 3.81 (m, 2H), 3.31 - 3.25 (m, 2H), 2.98 - 2.90 (m, 2H), 2.87 - 2.75 (m, 4H), 2.68 - 2.60 (m, 2H), 2.29 (s, 6H), 1.89 - 1.80 (m, 4H), 1.67 - 1.50 (m, 8H).

[0301] Example 80 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(5-methyl-4-oxo-4,5-dihydropyrazolo[1,5-a]pyrazine-2,6-diyl))bis(methylene))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 80) [ka] Following the procedure for compound 41, substituting L-serine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 80. LCMS (ESI): C 34 H 32 Calculated for Cl2N8O8; [M+H] + :751.58, Actual value:751.15. 1 H NMR (500 MHz, DMSO-d6) δ 7.83 (s, 2H), 7.73 - 7.66 (m, 2H), 7.51 (t, J = 7.6 Hz, 2H), 7.38 (s, 2H), 7.31 (d, J = 7.4 Hz, 2H), 7.26 (d, J = 1.7 Hz, 2H), 5.56 (s, 2H), 4.53 - 4.42 (m, 4H), 3.92 (s, 6H), 3.53 (s, 4H), 3.49 - 3.34 (m, 2H).

[0302] Example 81 Preparation of 2,2'-(2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-5-(((S)-pyrrolidin-3-yl)methyl)pyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 81) [ka] Following the procedure for compound 36, the title compound 81 was obtained by replacing (S)-tert-butyl 2-(hydroxymethyl)pyrrolidine-1-carboxylate (36A-1) with (S)-tert-butyl 3-(hydroxymethyl)pyrrolidine-1-carboxylate. LCMS (ESI): C 36 H 36 Calculated for Cl2N8O2 [M+H] + :683.2,685.2,Actual values:683.2,685.2. 1 H NMR (500 Mhz, DMSO-d6) δ 7.85 (s, 2H), 7.70 - 7.64 (m, 2H), 7.49 (t, J = 7.6 Hz, 2H), 7.39 (s, 2H), 7.26 (d, J = 7.4 Hz, 2H), 7.21 (d, J = 1.7 Hz, 2H), 5.54 (s, 2H), 4.50 - 4.41 (m, 2H), 4.26 - 4.18 (m, 4H), 3.84 - 3.46 (m, 4H), 3.12 - 3.06 (m, 4H), 2.27 (s, 6H), 2.18 - 2.09 (m, 2H).

[0303] Example 82 Preparation of 3,3'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(oxetane-3-carboxylic acid) (Compound 82) [ka] Following the procedure for compound 47, substituting 3-aminooxetane-3-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 82. LCMS (ESI): C 38 H 36 Calculated for Cl2N8O8; [M+H] + :803.65,Actual value:803.25. 1 H NMR (500 MHz, DMSO-d6) δ 7.88 (dd, J = 7.8, 1.8 Hz, 2H), 7.83 (s, 2H), 7.56 (q, J = 7.3 Hz, 2H), 7.45 (dd, J = 7.6, 1.8 Hz, 2H), 7.38 (s, 2H), 4.76 - 4.71 (s, 4H), 4.60 - 4.46 (m, 4H), 4.24 - 4.18 (m, 4H), 3.44 (s, 2H), 3.18 - 3.14 (m, 4H), 2.40 (s, 6H).

[0304] Example 83 Preparation of 2,2'-(2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-5-(2-(piperazin-1-yl)ethyl)pyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 83) [ka] Following the procedure for compound 47, substituting piperazine for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 83. LCMS (ESI): C 38 H 42 Cl2N 10 Calculated for O2; [M+H] + :741.3,743.3,Actual values:741.3,743.3. 1H NMR (500 MHz, DMSO-d6) δ 7.88 - 7.85 (m, 2H), 7.84 (s, 2H), 7.74 - 7.71 (m, 2H), 7.59 - 7.55 (m, 2H), 7.48 - 7.45 (m, 2H), 3.86 - 3.72 (m, 4H), 2.70 - 2.67 (m, 8H), 2.58 - 2.54 (m, 12H), 2.34 - 2.31 (m, 2H), 2.29 (s, 6H).

[0305] Example 84 Preparation of (2S,2'S,5S,5'S)-1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(5-hydroxypiperidine-2-carboxylic acid) (Compound 84) [ka] Following the procedure for compound 47, substituting (2S,5S)-5-hydroxypiperidine-2-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid gave the title compound 84. LCMS (ESI): C 42 H 44 Calculated for Cl2N8O8; [M+H] + :859.3,861.3,Actual values:859.3,861.3. 1 H NMR (500 MHz, DMSO-d6) δ 7.88 - 7.85 (m, 2H), 7.84 (s, 2H), 7.74 - 7.71 (m, 2H), 7.59 - 7.55 (m, 2H), 7.46 (s, 2H), 4.24 - 4.18 (m, 2H), 3.97 - 3.71 (m, 6H), 3.43 - 3.37 (m, 2H), 3.03 - 2.96 (m, 2H), 2.92 - 2.81 (m, 4H), 2.79 - 2.73 (m, 2H), 2.29 (s, 6H), 1.90 - 1.63 (m, 8H).

[0306] Example 85 Preparation of 1,1'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxycyclobutane-1-carboxylic acid) (Compound 85) [ka] Following the procedure for compound 47, substituting 1-amino-3-hydroxycyclobutane-1-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 85. LCMS (ESI): C 40 H 40 Calculated for Cl2N8O8; [M+H] + :831.71, Actual value:799.42. 1 H NMR (500 MHz, DMSO-d6) δ 7.89 (dd, J = 7.8, 1.8 Hz, 2H), 7.80 (d, J = 12.7 Hz, 2H), 7.56 - 7.45 (m, 6H), 7.36 (d, J = 2.6 Hz, 2H), 4.16 (s, 2H), 4.09 - 4.02 (m, 4H), 3.35 - 3.29 (m ,4H), 2.39 (m, 6H), 2.44 - 2.38 (m, 4H), 2.24- 2.20 (m, 4H).

[0307] Example 86 Preparation of 4,4'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(morpholine-3-carboxylic acid) (Compound 86) [ka] Following the procedure for compound 47, substituting morpholine-3-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 86. LCMS (ESI): C 40 H40 Calculated for Cl2N8O8; [M+H] + :830.23, Actual value:830.20. 1 H NMR (500 MHz, DMSO-d6) δ 7.89 (d, J = 7.8 Hz, 2H), 7.78 (s, 2H), 7.56 (t, J = 7.7 Hz, 2H), 7.44 (d, J = 7.6 Hz, 2H), 7.35 (s, 2H), 4.09 (t, J = 6.5 Hz, 4H), 3.85 - 3.53 (m, 10H), 3.24 - 2.76 (m, 8H), 2.40 (s, 6H).

[0308] Example 87 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(propane-1,2-diyl))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 87) [ka] To a stirred mixture of compound 29D (300 mg, 1.32 mmol, 1.0 equiv) in DMF (6 mL) was added CsCO (857 mg, 2.63 mmol, 2.0 equiv) and 1-bromopropan-2-one (288 mg, 2.1 mmol, 1.6 equiv) at 0 °C. The resulting mixture was stirred at 25 °C for 1 h. The reaction was quenched with water and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (100 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (2:1) to give compound 87A (300 mg, yield: 80.7%). LCMS (ESI): Calculated for C8H9BrN2O3; [M+H]+: 284.11, 286.11, Found: 284.11, 286.11.

[0309] To a stirred solution of compound 87A (200 mg, 0.709 mmol, 1.0 equiv) in DCM (10.00 mL) was added ethyl L-serine (234 mg, 1.76 mmol, 2.5 equiv) and 1 drop of HOAc at 25° C. After stirring at 25° C. for 0.5 h, sodium triacetoxyborohydride (895 mg, 4.21 mmol, 6.0 equiv) was added. The resulting mixture was stirred at 25° C. for 16 h. The reaction mixture was concentrated and purified by preparative liquid chromatography to give compound 87B (71 mg, yield: 25.1%). LCMS (ESI): C 15 H 21 Calculated for BrNO; [M+H]: 401.3, 403.3, Found: 401.3, 403.3.

[0310] Compound 87B (70 mg, 0.17 mmol, 2.2 equiv.), compound 1A (37 mg, 0.079 mmol, 1.0 equiv.), CsCO (129 mg, 0.40 mmol, 5 equiv.), and Pd(dppf)Cl·DCM (15 mg, 0.05 mmol, 0.30 equiv.) were dissolved in DMF (5.0 mL) and HO (1.0 mL) under a N atmosphere. The reaction was carried out at 90 °C for 2 h. After cooling, 30 mL of water was added, followed by extraction with 30 mL of EtOAc. The organic phase was washed with water and concentrated to give a residue. The reaction was purified by preparative HPLC to give compound 87 (6.9 mg, 9% yield). LCMS (ESI): C 38 H 40 Calculated for Cl2N8O8; [M+H]+: 807.7, 809.7, Found: 807.7, 809.7. 1H NMR (500 Mhz, DMSO-d6) δ 11.64 (s, 2H), 8.24 (s, 2H), 7.83 (s, 2H), 7.71 - 7.63 (m, 2H), 7.46 (t, J = 7.6 Hz, 2H), 7.35 (s, 2H), 7.30 (d, J = 7.4 Hz, 2H), 7.25 (d, J = 1.7 Hz, 2H), 5.58 (s, 2H), 4.50 - 4.41 (m, 4H), 4.30 - 4.19 (m, 4H), 4.01 - 3.87 (m, 2H), 3.79 - 3.67 (m, 6H), 3.54 - 3.23 (m, 2H), 2.21 (s, 6H).

[0311] Example 88 Preparation of 1,1'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(4-hydroxycyclohexane-1-carboxylic acid) (Compound 88) [ka] Following the procedure for compound 47, substituting 1-amino-4-hydroxycyclohexane-1-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 88. LCMS (ESI): C 44 H 48 Calculated for Cl2N8O8; [M+H] + :887.3,889.3,Actual values:887.3,889.3. 1H NMR (500 MHz, DMSO-d6) δ 7.88 - 7.85 (m, 2H), 7.84 (s, 2H), 7.74 - 7.71 (m, 2H), 7.59 - 7.54 (m, 2H), 7.46 (s, 2H), 4.25 - 4.21 (m, 2H), 3.83 - 3.79 (m, 4H), 3.76 - 3.70 (m, 2H), 3.50 - 3.47 (m, 2H), 2.87 - 2.71 (m, 4H), 2.29 (s, 6H), 1.95 - 1.85 (m, 4H), 1.82 - 1.69 (m, 8H), 1.62 - 1.52 (m, 4H).

[0312] Example 89 Preparation of (2S,2'S,3S,3'S)-1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(3-hydroxypyrrolidine-2-carboxylic acid) (Compound 89) [ka] Following the procedure for compound 47, substituting (2S,3S)-3-hydroxypyrrolidine-2-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 89. LCMS (ESI): C 40 H 40 Calculated for Cl2N8O8; [M+H] + :831.2,833.2,Actual values:831.2,833.2. 1H NMR (500 MHz, DMSO-d6) δ 7.88 - 7.85 (m, 2H), 7.84 (s, 2H), 7.74 - 7.71 (m, 2H), 7.59 - 7.54 (m, 2H), 7.46 (s, 2H), 4.97 - 4.93 (m, 2H), 4.17 - 4.10 (m, 2H), 3.98 - 3.77 (m, 4H), 3.30 (s, 2H), 3.04 - 2.93 (m, 2H), 2.91 - 2.74 (m, 6H), 2.29 (s, 6H), 1.89 - 1.72 (m, 4H).

[0313] Example 90 Preparation of 2,2'-(2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(5-(2-(((1r,3r)-3-hydroxycyclobutyl)amino)ethyl)-6-methylpyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 90) [ka] Following the procedure for compound 47, substituting (1R,3R)-3-aminocyclobutan-1-ol for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 90. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O4; [M+H] + :743.69, Actual value:743.53. 1 H NMR (500 MHz, DMSO-d6) δ 8.83 (s, 4H), 7.93 - 7.77 (m, 2H), 7.57 (t, J = 7.7 Hz, 2H), 7.46 (dd, J = 7.6, 1.8 Hz, 2H), 7.39 (s, 2H), 5.29 (s, 2H), 4.33 (m, 2H), 4.24 (m, 4H), 3.82 (d, J = 9.2 Hz, 2H), 3.21 - 3.08 (m, 4H), 2.40- 2.32 (m, 4H), 2.38 (d, J = 9.3 Hz, 6H), 2.11 - 1.99 (m, 4H).

[0314] Example 91 Preparation of 2,2'-(2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(5-(2-(((1s,3s)-3-hydroxycyclobutyl)amino)ethyl)-6-methylpyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 91) [ka] Following the procedure for compound 47, substituting (1S,3S)-3-aminocyclobutan-1-ol for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 91. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O4; [M+H] + :743.69, Actual value:743.51. 1 H NMR (500 MHz, DMSO-d6) δ 8.83 (s, 4H), 7.95 - 7.76 (m, 2H), 7.57 (t, J = 7.7 Hz, 2H), 7.46 (dd, J = 7.6, 1.7 Hz, 2H), 7.38 (s, 2H), 5.42 (s, 2H), 4.30 (m, 2H), 4.24 (t, J = 6.6 Hz, 4H), 3.91 (p, J = 7.3 Hz, 2H), 3.19 - 3.06 (m, 6.9 Hz, 4H), 2.38- 2.30 (m, 4H), 2.36 (s, 6H), 2.09 - 1.97 (m, 4H).

[0315] Example 92 Preparation of 2,2'-(2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(5-(((2S,4R)-4-hydroxypyrrolidin-2-yl)methyl)-6-methylpyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 92) [ka] Following the procedure for compound 29, substituting ((2S,4R)-4-hydroxypyrrolidin-2-yl)methyl 4-methylbenzenesulfonate for (S)-(5-oxopyrrolidin-2-yl)methyl 4-methylbenzenesulfonate gave the title compound 92. LCMS (ESI): C 36 H 36 Calculated for Cl2N8O4; [M+H] + :715.64, Actual value:715.24. 1 H NMR (500 MHz, DMSO-d6) δ 8.19 (m, 2H), 7.89 - 7.85 (m, 2H), 7.81 - 7.72 (m, 2H), 7.59 - 7.51 (m, 2H), 7.46 - 7.42 (m, 2H), 7.38 - 7.29 (m, 2H), 4.26 (s, 2H), 4.13 - 4.01 (m, 2H), 3.95 - 3.81 (m, 2H), 3.71 - 3.74 (m, 2H), 3.06 - 3.01 (m, 2H), 2.74 - 2.67 (m, 2H), 2.35 (m, 6H), 1.86 - 1.81 (m, 2H), 1.66 - 1.60 (m, 2H).

[0316] Example 93 Preparation of 2,2'-(2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(5-(2-(2-(2-hydroxyethyl)pyrrolidin-1-yl)ethyl)-6-methylpyrazolo[1,5-a]pyrazin-4(5H)-one) (Compound 93) [ka] Following the procedure for compound 47, substituting 2-(pyrrolidin-2-yl)ethan-1-ol for (S)-pyrrolidine-3-carboxylic acid gave the title compound 93. LCMS (ESI): C 42 H 48 Calculated for Cl2N8O4; [M+H] + :799.80, Actual value:799.12. 1H NMR (500 MHz, DMSO-d6) δ 7.89 (dd, J = 7.8, 1.8 Hz, 2H), 7.81 (s, 2H), 7.56 (t, J = 7.7 Hz, 2H), 7.45 (dd, J = 7.8, 1.8 Hz, 2H), 7.36 (s, 2H), 4.12 (s, 4H), 3.51 - 3.37 (m, 14H), 3.13 (s, 2H), 2.36 (s, 6H), 1.99 - 1.89 (m, 4H), 1.80 - 1.72 (m, 4H), 1.43 (s, 4H).

[0317] Example 94 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(propane-2,1-diyl))bis(azanediyl))bis(3-hydroxypropanoic acid) (Compound 94) [ka] To a stirred mixture of compound 29D (100 mg, 0.44 mmol, 1 equiv.) in THF (10 mL), compound 94A (82 mg, 0.88 mmol, 2.0 equiv.), PPh3 (173 mg, 0.66 mmol, 1.5 equiv.), and DIAD (133 mg, 0.66 mmol, 1.5 equiv.) were added at 0° C. under N2. The resulting mixture was stirred at 25° C. under N2 for 16 h. The reaction was quenched with water (10 mL) and extracted with EtOAc (10 mL×3). The combined organic layers were washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (1:1) to give compound 94B (80 mg, yield: 60.0%). LCMS (ESI): C 10 H 11 Calculated for BrClNO; [M+H]: 304.1, 306.1, Found: 304.1, 306.1.

[0318] To a stirred mixture of compound 94B (80 mg, 0.26 mmol, 1 equiv) in DMF (5.0 mL) was added KCO (72 mg, 0.52 mmol, 2.0 equiv) and HO (1.0 mL) at 25 °C. The resulting mixture was stirred at 60 °C for 2 h. After cooling, the reaction was quenched with water (30 mL) and extracted with EtOAc (10 mL × 3). The combined organic layer was washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give compound 94C (65 mg, yield: 86.6%). LCMS (ESI): C 10 H 12 Calculated for BrN3O2; [M+H]+: 286.01, 288.01, Found: 286.01, 288.01.

[0319] To a stirred mixture of compound 94C (65 mg, 0.22 mmol, 1.0 equiv) in DCM (5.0 mL) was added Dess-Martin periodate (186 mg, 0.44 mmol, 2.0 equiv) at 25 °C. The resulting mixture was stirred at 25 °C for 2 h. The reaction was quenched with water (30 mL) and extracted with DCM (10 mL × 3). The combined organic layer was washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give compound 94D (60 mg, yield: 92.7%). LCMS (ESI): C 10 H 10 Calculated for BrN3O2; [M+H]+: 284.01, 286.01, Found: 284.01, 286.01.

[0320] To a stirred mixture of compound 94D (60 mg, 0.20 mmol, 1.0 equiv) in DMA (5.0 mL) was added ethyl L-serine hydrochloride (136 mg, 0.8 mmol, 4.0 equiv), DIEA (103 mg, 0.8 mmol, 4.0 equiv), and 1 drop of HOAc at 25° C. After stirring for 0.5 h, sodium triacetoxyborohydride (339 mg, 1.6 mmol, 8.0 equiv) was added, and the resulting mixture was stirred at 25° C. for 12 h. The reaction was quenched with water (30 mL) and extracted with EtOAc (15 mL×3). The combined organic layer was washed with brine (20 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give compound 94E (30 mg, yield: 35.4%). LCMS (ESI): C 15 H 21 Calculated for BrN4O4; [M+H]+: 401.1, 403.1, Found: 401.1, 403.1.

[0321] To a stirred mixture of compound 1A (17 mg, 0.04 mmol, 1 equiv.) in DMF (5.0 mL), compound 94E (30 mg, 0.08 mmol, 2.1 equiv.), Pd(dppf)Cl DCM (6.0 mg, 0.01 mmol, 0.20 equiv.), CsCO (58 mg, 0.20 mmol, 5.0 equiv.), and HO (1.0 mL) were added at 25 °C. The resulting mixture was stirred at 80 °C under N for 2 h. After cooling to 25 °C, aqueous NaOH (2.0 M, 1.0 mL) was added, and the resulting mixture was stirred at 60 °C for 1 h. After cooling to 25 °C, the resulting mixture was filtered. The filtrate was collected and purified by preparative HPLC to give compound 94 (5.0 mg, yield: 17%). LCMS (ESI): C 38 H 40 Calculated for Cl2N8O8; [M+H]+: 807.2, 809.2, Found: 807.2, 809.2. 1H NMR (500 MHz, DMSO-d6) δ 7.88 - 7.85 (m, 2H), 7.84 (s, 2H), 7.74 - 7.71 (m, 2H), 7.59 - 7.55 (m, 2H), 7.46 - 7.44 (m, 2H), 4.95 - 4.81 (m, 4H), 4.48 - 4.36 (m, 2H), 3.67 - 3.39 (m, 6H), 2.97 - 2.83 (m, 4H), 2.28 (s, 6H), 1.32 (s, 6H).

[0322] Example 95 Preparation of (2S,2'S)-2,2'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxy-2-methylpropanoic acid) (Compound 95) [ka] Following the procedure for compound 47, substituting (S)-2-amino-3-hydroxy-2-methylpropanoic acid for (S)-pyrrolidine-3-carboxylic acid gave the title compound 95. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O8; [M+H] + :807.69, Actual value:807.01. 1 H NMR (500 MHz, DMSO-d6) δ 8.06 (s, 2H), 7.90 - 7.82 (m, 4H), 7.57 (t, J = 7.7 Hz, 2H), 7.46 (dd, J = 7.6, 1.7 Hz, 2H), 7.38 (s, 2H), 4.33 - 4.26 (t, J = 7.6 Hz, 4H), 3.89 (s, 2H), 3.77-7.73 (m, 4H), 3.62 - 3.55 (m, 4H), 3.17 (d, J = 7.8 Hz, 6H), 2.38 (s, 6H). (s, 6H).

[0323] Example 96 Preparation of (2R,2'R)-2,2'-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxy-2-methylpropanoic acid) (Compound 96) [ka] Following the procedure for compound 47, substituting (R)-2-amino-3-hydroxy-2-methylpropanoic acid for (S)-pyrrolidine-3-carboxylic acid gave the title compound 96. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O8; [M+H] + :807.69, Actual value:807.01. 1 H NMR (500 MHz, DMSO-d6) δ 8.06 (s, 2H), 7.90 - 7.82 (m, 4H), 7.57 (t, J = 7.7 Hz, 2H), 7.46 (dd, J = 7.6, 1.7 Hz, 2H), 7.38 (s, 2H), 4.33 - 4.26 (t, J = 7.6 Hz, 4H), 3.89 (s, 2H), 3.77-7.73 (m, 4H), 3.62 - 3.55 (m, 4H), 3.17 (d, J = 7.8 Hz, 6H), 2.38 (s, 6H). (s, 6H).

[0324] Example 97 Preparation of (1s,1's,3s,3's)-((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(cyclobutane-3,1-diyl)diacetate (Compound 97) [ka] Following the procedure for compound 47, substituting (1S,3S)-3-aminocyclobutylacetic acid hydrochloride for (S)-pyrrolidine-3-carboxylic acid gave the title compound 97. LCMS (ESI): C 42 H 46 Calculated for Cl4N8O6; [M+H] + :900.68, Actual value:828.12. 1 H NMR (500 MHz, DMSO-d6) δ 8.81 (s, 4H), 7.93 - 7.74 (m, 2H), 7.55 (t, J = 7.7 Hz, 2H), 7.44 (dd, J = 7.6, 1.7 Hz, 2H), 7.36 (s, 2H), 5.40 (s, 2H), 4.28 (m, 2H), 4.22 (t, J = 6.6 Hz, 4H), 3.90 (p, J = 7.3 Hz, 2H), 3.18 - 3.05 (m, 6.9 Hz, 4H), 2.36- 2.29 (m, 4H), 2.35 (s, 6H), 2.06 - 1.94 (m, 4H), 2.02 (s, 6H).

[0325] Example 98 Preparation of ((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(ethane-2,1-diyl) diacetate (Compound 98) [ka] Following the procedure for compound 47, substituting ethyl 2-aminoacetate hydrochloride for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 98. LCMS (ESI): C 38 H 40 Calculated for Cl2N8O6; [M+H] + :775.2,777.2,Actual values:775.2,777.2. 1H NMR (500 MHz, DMSO-d6): δ 7.88 - 7.85 (m, 2H), 7.84 (s, 2H), 7.75 - 7.71 (m, 2H), 7.59 - 7.54 (m, 2H), 7.48 - 7.46 (m, 2H), 4.14 - 4.08 (m, 4H), 3.83 - 3.76 (m, 4H), 3.07 - 3.01 (m, 4H), 2.86 - 2.81 (m, 2H), 2.76 - 2.72 (m, 4H), 2.29 (s, 6H), 2.02 (s, 6H).

[0326] Example 99 Preparation of diethyl 1,1'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxycyclobutane-1-carboxylate) (Compound 99) [ka] Following the procedure for compound 47, substituting ethyl 1-amino-3-hydroxycyclobutane-1-carboxylate for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 99. LCMS (ESI): C 44 H 48 Calculated for Cl2N8O8; [M+H] + :887.3,889.3,Actual values:887.3,889.3. 1H NMR (500 MHz, DMSO-d6) δ 7.89 (dd, J = 7.8, 1.8 Hz, 2H), 7.81 (d, J = 12.7 Hz, 2H), 7.56 - 7.44 (m, 6H), 7.36 (d, J = 2.6 Hz, 2H), 4.17 (s, 2H), 4.12 (m, 4H), 4.08 - 4.02 (m, 4H), 3.34 - 3.29 (m,4H), 2.39 (m, 6H), 2.44 - 2.38 (m, 4H), 2.24- 2.20 (m, 4H), 1.24(d, J = 10Hz, 6H).

[0327] Example 100 Preparation of diethyl 2,2'-(((((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-1-oxopyrrolo[1,2-a]pyrazine-7,2(1H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))(2S,2'S)-bis(3-hydroxypropanoate) (Compound 100) [ka] Following the procedure of Example 48, substituting L-ethyl serine for (S)-pyrrolidin-3-ol, gave the title compound 100. LCMS (ESI): C 42 H 46 Calculated for Cl2N6O8; [M+H] + :833.3,835.3,Actual values:833.3,835.3. 1H NMR (500 MHz, DMSO-d6) δ 7.76 (d, J = 1.8 Hz, 2H), 7.67 (dd, J = 7.8, 1.7 Hz, 2H), 7.46 (t, J = 7.7 Hz, 2H), 7.28 (ddd, J = 5.0, 3.7, 1.7 Hz, 4H), 7.18 (d, J = 1.7 Hz, 2H), 4.83 (s, 2H), 4.07 (q, J = 7.1 Hz, 4H), 3.99 - 3.91 (m, 4H), 3.58 - 3.52 (m, 4H), 2.82 (dt, J = 11.7, 6.9 Hz, 2H), 2.70 - 2.63 (m, 2H), 2.52 (d, J = 1.9 Hz, 2H), 2.27 (s, 6H), 1.16 (d, J = 7.2 Hz, 6H).

[0328] Example 101 Preparation of (1r,1'r,3r,3'r)-1,1'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxycyclobutane-1-carboxylic acid) (Compound 101) [ka] Following the procedure of Example 47, substituting (1R,3R)-1-amino-3-hydroxycyclobutane-1-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 101. LCMS (ESI): C 40 H 40 Calculated for Cl2N8O8; [M+H]+: 831.2, 833.2, Found: 831.2, 833.2. 1H NMR (500 MHz, DMSO-d6) δ 7.89 - 7.85 (m, 2H), 7.84 (s, 2H), 7.75 - 7.71 (m, 2H), 7.59 - 7.54 (m, 2H), 7.48 - 7.46 (m, 2H), 5.02 - 4.98 (m, 2H), 4.80 - 4.77 (m, 2H), 3.92 - 3.84 (m, 2H), 3.83 - 3.78 (m, 4H), 2.87 - 2.73 (m, 4H), 2.68 - 2.62 (m, 4H), 2.29 (s, 6H), 2.24 - 2.19 (m, 4H).

[0329] Example 102 Preparation of (1S,1'S,3S,3'S)-1,1'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))bis(3-hydroxycyclobutane-1-carboxylic acid) (Compound 102) [ka] Following the procedure of Example 47, substituting (1S,3S)-1-amino-3-hydroxycyclobutane-1-carboxylic acid for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 102. LCMS (ESI): C 40 H 40 Calculated for Cl2N8O8; [M+H]+: 831.2, 833.2, Found: 831.2, 833.2. 1H NMR (500 MHz, DMSO-d6) δ 7.89 - 7.85 (m, 2H), 7.84 (s, 2H), 7.75 - 7.71 (m, 2H), 7.59 - 7.54 (m, 2H), 7.48 - 7.46 (m, 2H), 5.02 - 4.98 (m, 2H), 4.80 - 4.77 (m, 2H), 3.92 - 3.84 (m, 2H), 3.83 - 3.78 (m, 4H), 2.87 - 2.73 (m, 4H), 2.68 - 2.62 (m, 4H), 2.29 (s, 6H), 2.24 - 2.19 (m, 4H).

[0330] Example 103 Preparation of diethyl 1,1'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))(1R,1'R,3R,3'R)-bis(3-hydroxycyclobutane-1-carboxylic acid) (Compound 103) [ka] Following the procedure of Example 47, substituting ethyl (1R,3R)-1-amino-3-hydroxycyclobutane-1-carboxylate for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 103. LCMS (ESI): C 44 H 48 Calculated for Cl2N8O8; [M+H]+: 887.3, 889.3, Found: 887.3, 889.3. 1H NMR (500 MHz, DMSO-d6) δ 7.95 - 7.88 (m, 2H), 7.77 (s, 2H), 7.60 - 7.54 (m, 2H), 7.49 - 7.42 (m, 2H), 7.37 (s, 2H), 5.12 - 5.05 (m, 2H), 4.12 - 4.02 (m, 6H), 4.01 - 3.94 (m, 4H), 2.75 - 2.61 (m, 6H), 2.41 (s, 6H), 2.10 - 1.98 (m, 8H), 1.19 - 1.13 (m, 6H).

[0331] Example 104 Preparation of diethyl 1,1'-((((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(6-methyl-4-oxopyrazolo[1,5-a]pyrazine-2,5(4H)-diyl))bis(ethane-2,1-diyl))bis(azanediyl))(1S,1'S,3S,3'S)-bis(3-hydroxycyclobutane-1-carboxylic acid) (Compound 104) [ka] Following the procedure of Example 47, substituting (1S,3S)-1-amino-3-hydroxycyclobutane-1-carboxylate for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 104. LCMS (ESI): C 44 H 48 Calculated for Cl2N8O8; [M+H]+: 887.3, 889.3, Found: 887.3, 889.3. 1 H NMR (500 MHz, DMSO-d6) δ 7.95 - 7.88 (m, 2H), 7.77 (s, 2H), 7.60 - 7.54 (m, 2H), 7.49 - 7.42 (m, 2H), 7.37 (s, 2H), 5.12 - 5.05 (m, 2H), 4.12 - 4.02 (m, 6H), 4.01 - 3.94 (m, 4H), 2.75 - 2.61 (m, 6H), 2.41 (s, 6H), 2.10 - 1.98 (m, 8H), 1.19 - 1.13 (m, 6H).

[0332] Example 105 Preparation of diethyl 1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(3-methyl-1-oxopyrrolo[1,2-a]pyrazine-7,2(1H)-diyl))bis(ethane-2,1-diyl))(3S,3'S)-bis(pyrrolidine-3-carboxylic acid) (Compound 105) [ka] Following the procedure of Example 48, substituting (S)-ethyl pyrrolidine-3-carboxylate for (S)-pyrrolidin-3-ol, gave the title compound 105. LCMS (ESI): C 46 H 50 Calculated for Cl2N6O6; [M+H] + :853.84, Actual value:853.43. 1 H NMR (500 MHz, DMSO-d6) δ 7.77 (d, J = 1.8 Hz, 2H), 7.67 (dd, J = 7.8, 1.7 Hz, 2H), 7.47 (t, J = 7.6 Hz, 2H), 7.32 - 7.26 (m, 4H), 7.18 (d, J = 1.7 Hz, 2H), 4.07 (q, J = 7.1 Hz, 4H), 3.98 (t, J = 7.1 Hz, 4H), 3.04 - 2.95 (m, 2H), 2.81 (t, J = 8.8 Hz, 2H), 2.69 (dd, J = 9.2, 6.1 Hz, 2H), 2.65 - 2.55 (m, 8H), 2.27 (s, 6H), 1.99 - 1.90 (m, 4H), 1.18 (t, J = 7.1 Hz, 6H).

[0333] Example 106 Preparation of diethyl 1,1'-(((2,2'-dichloro[1,1'-biphenyl]-3,3'-diyl)bis(2-methyl-1-oxo-1,2-dihydropyrrolo[1,2-a]pyrazine-7,3-diyl))bis(methylene))(3S,3'S)-bis(pyrrolidine-3-carboxylic acid) (Compound 106) [ka] Following the procedure of Example 43, substituting (S)-ethyl pyrrolidine-3-carboxylate for (S)-pyrrolidine-3-carboxylic acid, gave the title compound 106. LCMS (ESI): C 44 H 46 Calculated for Cl2N6O6; [M+H] + :825.79, Actual value:825.51. 1 H NMR (500 MHz, DMSO-d6) δ 7.81 (s, 2H), 7.71 - 7.65 (m, 2H), 7.50 (t, J = 7.6 Hz, 2H), 7.38 (s, 2H), 7.31 (d, J = 7.4 Hz, 2H), 7.26 (d, J = 1.7 Hz, 2H), 4.05 (q, J = 7.1 Hz, 4H), 3.94 (s, 6H), 3.55 (s, 4H), 3.21 - 3.08 (m, 4H), 3.04 - 2.93 (m, 4H), 2.63 - 2.55 (m, 2H), 2.04 - 1.94 (m, 2H), 1.90 - 1.77 (m, 2H), 1.14 (t, J = 7.1 Hz, 6H).

[0334] Biological Examples Example 107 PD-1 / PD-L1 Homogeneous Time-Resolved Fluorescence (HTRF) Binding Assay Assays were performed in standard black 384-well polystyrene plates with a final volume of 20 μL. Inhibitors were first serially diluted in DMSO and then added to the plate wells prior to the addition of other reaction components. The final concentration of DMSO in the assay was 1%. Assays were performed at 25°C in PBS buffer (pH 7.4) containing 0.05% Tween-20 and 0.1% BSA. Recombinant human PD-L1 protein (19-238) with a C-terminal His tag was purchased from AcroBiosystems (PD1-H5229). Recombinant human PD-1 protein (25-167) with a C-terminal Fe tag was also purchased from AcroBiosystems (PD1-H5257). PD-L1 and PD-1 proteins were diluted in assay buffer, and 10 μL was added to the plate wells. The plate was centrifuged, and the proteins were pre-incubated with the inhibitors for 40 minutes. Following incubation, 10 μL of HTRF detection buffer supplemented with Fe-specific europium cryptate-labeled anti-human IgG (PerkinElmer-AD0212) and anti-His antibody conjugated to SureLight® allophycocyanin (APC, PerkinElmer-AD0059H) was added. After centrifugation, the plate was incubated at 25°C for 60 minutes and then read on a PHERAstar FS plate reader (665 nm / 620 nm ratio). Final concentrations in the assay were 3 nM PD1, 10 nM PD-L1, 1 nM europium anti-human IgG, and 20 nM anti-His-allophycocyanin. IC was determined by curve fitting of percent control activity versus the logarithm of inhibitor concentration. 50 Measurements were carried out. [Table 2] A: Activity<10nM, B: 10nM≦Activity<100nM, C: Activity≧100nM

[0335] Example 108 PD-L1 Dimerization To determine whether the compounds could specifically dimerize the extracellular domain of PD-L1, the compounds were tested in a biochemical protein-protein interaction assay.

[0336] (1) For each column of 10 samples, compounds were serially diluted 1:3 in DMSO.

[0337] (2) Using Echo, 0.2 μL of compound solution from each row was transferred to a 384-well assay plate, with each column containing two replicates.

[0338] (3) 20 μL of the prepared mixture containing PD-L1-Eu and PD-L1-A2 solutions was added to the assay plate and centrifuged at 1000 rpm for 1 minute.

[0339] (4) Incubated at 25°C for 120 minutes.

[0340] (5) The fluorescent signal was read using an Envision 2104 plate reader.

[0341] (6) The ratio (665 nm / 615 nm) signal was read on Envision.

[0342] (7) The raw data were analyzed using equation (V. Data Analysis). [Table 3] A: EC50<50nM, B:50nM≦EC50<100nM, C:EC50≧100nM

[0343] Example 109 PDL1 Jurkat-NFAT Reporter Assay Preparation of Hep3B-OS8-hPDL1 1. Hep3B-OS8-hPDL1 cells were cultured in 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin, and also containing 100 μg / mL of G418 and hygromycin B.

[0344] 2. Resuspend the cells in RPMI 1640 medium containing 10% FBS to a cell density of 1.25 x 10 5 Adjusted to cells / mL.

[0345] 3. Cells were seeded into 96-well flat-bottom plates (1.25x10 4 cells / 100 μL / well).

[0346] b. Preparation of compound solutions 4. The medium was removed from the pre-plated Hep3B-OS8-PDL1 cells and washed once with 200 μL of assay medium.

[0347] 5. Compound dilutions were prepared in RPMI 1640 medium containing 10% FBS according to the layout.

[0348] 6. Nine concentrations of compound (3, 1, 0.3, 0.1, 0.03, 0.01, 0.003, 0.001, and 0.0003 μM) were added to each well in a volume of 50 μL. Keytruda at a concentration of 5 μg / mL was included as a positive control.

[0349] 7. Incubate at 37°C and 5% CO2 for 20-30 minutes.

[0350] c. Preparation of Jurkat-NFAT-PD1 8. Jurkat-NFAT-PD1 cells were cultured in 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin, and also supplemented with 1000 μg / mL hygromycin B and 0.3 μg / mL puromycin.

[0351] 9. On the second day of the assay, resuspend the cells in RPMI 1640 medium containing 10% FBS to a cell density of 2.5 x 10 5 Adjusted to cells / mL.

[0352] 10. Cells were seeded into 96-well flat-bottom plates (1.25x10 4 cells / 50 μL / well).

[0353] 11. The assay plate was incubated in a humidified 37°C, 5% CO2 incubator for 6 hours.

[0354] 12. Cultured cells were allowed to equilibrate at room temperature for 5-10 minutes.

[0355] 13. An equal volume (100 μL / well) of ONE-Glo™ Luciferase Assay System was added to each well, and the cells were allowed to lyse completely for at least 3 minutes before being measured in a luminometer. [Table 4] A: EC50<500nM, B: EC50≧500nM

[0356] Example 110 Co-culture assay of Hep3B-OS8-hPDL1 and T cells Tumor preparation 1. Hep3B-OS8-hPDL1 cells were cultured in 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin, and also containing 100 μg / mL of G418 and hygromycin B.

[0357] 2. Hep3B-OS8-hPDL1 cells were harvested and treated with 10 μg / mL mitomycin C at 37°C for 1.5 hours, after which the cells were thoroughly washed four times with PBS.

[0358] 3. Resuspend the cells in RPMI 1640 medium containing 10% FBS to a cell density of 5 x 10 5 Adjusted to cells / mL.

[0359] 4. Cells were seeded into 96-well flat-bottom plates (2.5x10 4 cells / 50 μL / well).

[0360] b. CD3+ T cell isolation (30 mL blood) 5. Human blood samples from individual donors were diluted with an equal volume of sterile PBS, for example, 25 mL of sterile PBS was added to 25 mL of fresh whole blood and mixed thoroughly by gentle shaking.

[0361] 6.15 mL of Lymphoprep medium was transferred to a new 50 mL centrifuge tube.

[0362] 7. Add the diluted blood sample as gently as possible to the surface of the Ficoll medium, ensuring that there is a clear boundary between the two liquids, and ensure that the volume ratio of Ficoll to diluted blood (30 mL) is 1:2.

[0363] 8. The tube was gently agitated and centrifuged at 1000 x g for 25 minutes at 20°C with acceleration (5) and minimum deceleration (0) during centrifugation.

[0364] 9. After centrifugation, a total of four interfaces were observed: plasma, mononuclear cells, Ficoll medium, and RBC layers. The tube was then moved as gently as possible to keep the four interfaces separate. The second layer of mononuclear cells was carefully aspirated and transferred to a new sterile centrifuge tube. If necessary, a certain amount of plasma was aspirated instead of the Ficoll medium.

[0365] 10. Sterile PBS in an amount three times the volume of PBMC was added to the tube containing PBMC.

[0366] After washing the cells twice with 11.5-10 mL of PBS, the cells were counted using a cytometer. They were centrifuged at 350 × g for 10 minutes at 20 °C with an acceleration and deceleration setting of 5.

[0367] 12. Resuspend the cells in the recommended medium to a final concentration of 5x10 PBMCs. 7 Adjusted to cells / mL.

[0368] 13. CD3+ T cells were isolated using the EasySep™ Human T Cell Isolation Kit (STEMCELL Technologies #17951) and seeded into 96-well flat-bottom plates (5 x 10 4 cells / 100 μL / well).

[0369] c. Preparation of compound solutions 14. Compound dilutions were prepared in RPMI 1640 medium containing 10% FBS according to the layout. 15. Compounds were added to each well in a volume of 50 μL (three compounds (GLC01-258, GLC01-269, and GLC01-465) at seven concentrations (0.03, 0.1, 0.3, 1, 3, 10, and 30 μM), and six compounds (GLC01-411, GLC01-292, GLC01-445, GLC01-475, GLC01-470, and GLC01-468) at the same concentration (1 μM).

[0370] 16. Keytruda at a concentration of 5 μg / mL was included as a positive control.

[0371] Incubated at 17.37°C and 5% CO2 for 72 hours.

[0372] 18. The supernatant was collected by centrifugation and IFN-γ was measured by ELISA. [Table 5] A: EC50<50nM, B:50nM≦EC50<100nM, C:EC50≧100nM

[0373] Example 111 Mouse PK Studies (1) Compounds were weighed and dissolved in a vehicle of 5% saline solution at 1 mg / mL, shaken vigorously, and sonicated to form a clear, colorless solution, which was orally administered at a dose of 10 mg / kg to groups of three overnight-fasted mice.

[0374] (2) Blood samples were collected from the submandibular vein using sodium heparin for anticoagulation. After collection, the blood was placed on ice and centrifuged within 1 hour to separate the plasma (centrifugation conditions: 8000 rpm, 6 minutes, 2-8°C). Blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours.

[0375] (3) The samples were stored in a -20°C freezer. 160 μL of ice-cold acetonitrile containing the internal standard was added to the plasma sample (40 μL), vortexed for 3 minutes, and centrifuged at 11,000 rpm for 5 minutes. 100 μL of the supernatant was added to 100 μL of water, and 5 μL of the supernatant was injected into the LC / MS / MS system to detect the compound (if the compound was an ester, the acid was detected). The results are shown in Table 6. [Table 6] A: Cmax ≥ 1000 ng / mL B: 100ng / mL≦Cmax<1000ng / mL C: Cmax<100ng / mL D:AUC last ≥ 2000 ng*h / mL E: 500ng*h / mL≦AUC last <2000ng*h / mL F:AUC last <500 ng*h / mL

[0376] While various embodiments have been described above, it should be understood that such disclosure is presented by way of example only, and not limitation. Thus, the breadth and scope of the subject compositions and methods should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0377] The above description is intended to teach those skilled in the art how to practice the present application and is not intended to detail all obvious modifications and variations that will become apparent to those skilled in the art upon reading the present description. However, all such obvious modifications and variations are intended to be included within the scope of the present application as defined by the claims. The claims are intended to cover any sequence of components and steps that is effective to achieve the intended purpose, unless the context specifically indicates to the contrary.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, solvate, or tautomer thereof. 【Chemistry 1】 (In the formula, （１） Each of A and B is a halogen, a cyano, or a —N ３ independently selected from the group consisting of alkyl and substituted alkyl, amine, alkylamine, and alkoxy; Z １ is =N-, -N(R １ ) -, = C (R ２ )- or -S-; Z ２ is =N-, -N(R １ ) -, = C (R ２ )- or -S-; Z ６ is =N-, -N(R １ ) -, = C (R ２ )- or -S-; Z ７ is =N-, -N(R １ ) -, = C (R ２ )- or -S-; Each R １ is independently —H, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, or aryl, and each R ２ is independently halogen, cyano, cycloalkyl, substituted alkyl, alkenyl, alkynyl, or aryl; Z ３ is a nitrogen atom or a carbon atom, Z ４ is -N=, -N(R ３ ) -, = C (R ４ ) - or -C(R ５ R ６ ) - and Z ５ is a nitrogen atom or a carbon atom, Z ８ is a nitrogen atom or a carbon atom, Z ９ is -N=, -N(R ３ ) -, = C (R ４ ) - or -C(R ５ R ６ ) - and Z １０ is a nitrogen atom or a carbon atom, Each R ３ is independently —H, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, or aryl; R ４ , R ５ and R ６ each is independently halogen, cyano, cycloalkyl, substituted alkyl, alkenyl, alkynyl, or aryl; L １ and L ３ each independently represents a ring 3 and a ring W １ Between ring 6 and W ３ and wherein the alkyl, the substituted alkyl, or the heteroatom chain each contains m atoms, where m=0, 1, 2, 3, 4, 5, or 6; and when m is 0, W １ Or W ３ are each directly attached to the corresponding nitrogen atom in ring 3 or ring 6, L2 and L ４ each independently represents a ring 3 and a ring W ２ Between ring 6 and W ４ and wherein the alkyl, the substituted alkyl, or the heteroatom chain each contains m atoms, where m=0, 1, 2, 3, 4, 5, or 6; and when m is 0, W ２ Or W ４ are each directly attached to the corresponding nitrogen atom in ring 3 or ring 6, One or more of W 1 , W 2 , W 3 , and W 4 have the general formula: 【Chemistry 2】 wherein R 7 is alkyl or substituted alkyl and R 8 is —H, alkyl, or substituted alkyl; or （２） each of A and B is independently —Cl or methyl; Z １ is =N-, -N(R １ ) -, = C (R ２ )- or -S-; Z ２ is =N-, -N(R １ ) -, = C (R ２ )- or -S-; Z ６ is =N-, -N(R １ ) -, = C (R ２ )- or -S-; Z ７ is =N-, -N(R １ ) -, = C (R ２ )- or -S-; Each R １ is independently —H, alkyl, cycloalkyl, or substituted alkyl, and each R ２ are independently —H, —F, or methyl; Z ３ is a nitrogen atom or a carbon atom, Z ４ is -N=, -N(R ３ ) -, = C (R ４ ) - or -C(R ５ R ６ ) - and Z ５ is a nitrogen atom or a carbon atom, Z ８ is a nitrogen atom or a carbon atom, Z ９ is -N=, -N(R ３ ) -, = C (R ４ ) - or -C(R ５ R ６ ) - and Z １０ is a nitrogen atom or a carbon atom, Each R ３ is independently —H, alkyl, cycloalkyl, or substituted alkyl; R ４ , R ５ and R ６ each is independently halogen, cyano, cycloalkyl, or substituted alkyl; L １ and L ３ each independently represents a ring 3 and a ring W １ Between ring 6 and W ３ and wherein the alkyl, the substituted alkyl, or the heteroatom chain each contains m atoms, where m=0, 1, 2, 3, 4, 5, or 6; and when m is 0, W １ Or W ３ are each directly attached to the corresponding nitrogen atom in ring 3 or ring 6, L2 and L ４ each independently represents a ring 3 and a ring W ２ Between ring 6 and W ４ and wherein the alkyl, the substituted alkyl, or the heteroatom chain each contains m atoms, where m=0, 1, 2, 3, 4, 5, or 6; and when m is 0, W ２ Or W ４ are each directly attached to the corresponding nitrogen atom in ring 3 or ring 6, One or more of W 1 , W 2 , W 3 , and W 4 have the general formula: 【Transformation 3】 wherein R 7 is alkyl or substituted alkyl, and R 8 is —H, alkyl, or substituted alkyl.

2. 2. The compound of claim 1, wherein rings 2 and 3, and / or rings 5 ​​and 6 of the compound are independently selected from the group consisting of: 【Chemistry 4】

3. 2. The compound of claim 1, wherein rings 2 and 3, and / or rings 5 ​​and 6 of the compound are independently selected from the group consisting of: 【Transformation 5】

4. L １ , L ２ , L ３ and L ４ Each of these is independently C １ ~C ３ The compound of any one of claims 1 to 3, which is alkyl.

5. R ７ is one of the following: 【Transformation 6】 R ８ The compound of claim 1 , wherein: is independently —H, alkyl, or substituted alkyl.

6. R ７ is one of the following: 【Transformation 7】 R ８ The compound of claim 1 , wherein: is independently —H, alkyl, or substituted alkyl.

7. A compound selected from the group consisting of: 【Transformation 8】 【change】 【change】 【change】 【change】 【change】 【change】

8. 8. The compound of claim 7, wherein the compound is selected from the group consisting of: 【Chemistry 9】 【change】

9. The compound has the formula 【Chemistry 10】 and or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, deuterated compound, or tautomer thereof.

10. The compound has the formula 【Chemistry 11】 and or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, deuterated compound, or tautomer thereof.

11. The compound has the formula 【Chemistry 12】 and or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, deuterated compound, or tautomer thereof.

12. A compound according to any one of claims 1 to 11; and a pharmaceutically acceptable carrier.

13. 13. Use of the pharmaceutical composition of claim 12 in the preparation of a medicament for the treatment of a disease associated with PD-L1.

14. The use according to claim 13, wherein the disease is cancer.

15. The use according to claim 13, wherein the disease is an immune-related disease.

16. The use according to claim 13, wherein the disease is an infectious disease.

17. 17. The use according to claim 16, wherein the infection is a hepatitis B virus infection.

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