Bifunctional degraders and uses thereof

Abstract

Provided herein a compound and compositions that bind to p62 and a target of interest, and promote degradation of the target of interest. In some embodiments, a compound described herein is a compound of formula I: A-B-C (I) or a pharmaceutically acceptable salt thereof, wherein A is an agent that binds or associated with p62; B is a linker moiety; and C is a target binding moiety.

Description

BIFUNCTIONAL DEGRADERS AND USES THEREOF BACKGROUND

[0001] p62 is a multifunctional protein involved in the lysosomal degradation of ubiquinated proteins via the autophagy pathway. Liu WJ, et al., Cell Mol Biol Lett.2016 Dec 13;21:29. The ubiquitin-proteasome system (UPS) and autophagy are “two major degradative pathways of proteins in eukaryotic cells.” Shin WH, et al., BMB Rep.2020 Jan;53(1):56-63. These systems are integral to cell survival in both normal conditions and under stress. Liu WJ, et al., Cell Mol Biol Lett.2016 Dec 13;21:29. SUMMARY

[0002] p62 is an autophagy substrate that has been shown to deliver ubiquitinated proteins to the lysosome for degradation. Liu WJ, et al., Cell Mol Biol Lett. 2016 Dec 13;21:29. The therapeutic potential of harnessing cellular degradation systems to direct removal of specific targets has been well appreciated and extensively investigated for more than a decade. See, for example, Bondeson & Crews Annu Rev Pharmacol Toxicol 57:107-123 (Sept. 6, 2017), and references cited therein. p62 contributes to the autophagy process by interacting with ubiquitin or polyubiquitin chains on certain cargo, and then delivering the cargo to the lysosome for degradation. An overview of the relationship between p62 and the autophagy process is described by Liu WJ, et al., in Cell Mol Biol Lett.2016 Dec 13;21:29.

[0003] The present application provides, among other things, new compounds and uses thereof that utilize p62 to activate autophagy of cargo or target of interest. Without being bound by theory, it is understood that compounds described herein bind to both p62 and a target of interest, thereby causing p62 to deliver the cargo of interest to the the lysosome and thereby begin the degradation process of the target of interest.

[0004] In some embodiments, the present disclosure provides a compound formula I: A-B-C I or a pharmaceutically acceptable salt thereof, wherein A is a moiety that binds or associates with p62;B is a linker moiety; and C is a target binding moiety.

[0005] In some embodiments, a compound of formula I is a compound of formula II-1 or II- 2:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, - ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa;B is a linker moiety; and C is a target binding moiety.

[0006] In some embodiments, a compound of formula I is a compound of formula III-1 or III- 2:or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa;B is a linker moiety; and C is a target binding moiety.

[0007] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound described herein, and a pharmaceutically acceptable carrier, filler, or diluent.

[0008] In some embodiments, the present disclosure provides a method of treating a disease, disorder, or condition in a subject comprising administering to the subject a compound described herein.

[0009] In some embodiments, the present disclosure provides a method of inducing degradation of a target in a biological sample, comprising contacting the biological sample with a compound described herein.

[0010] In some embodiments, the present disclosure provides compounds that bind to p62. In some embodiments, a compound that binds p62 is a compound is of formula X:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa.

[0011] In some embodiments, a compound that binds p62 is a compound is of formula XI:or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic;each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa. BRIEF DESCRIPTION OF THE DRAWING

[0012] FIG.1 is a bar graph illustrating degradation levels of NSCLC-associated EML4-ALK fusion with agents and inactive controls.

[0013] FIG. 2 is a bar graph illustrating dose dependent, compound mediated cellular co- localization of endogenous p62 and endogenous ALK at 1 hour, 6 hours, and 24 hours. Co- treatment with an early (PIK-III) or late (BafA1) autophagy inhibitor sustains p62 and ALK co- localization out to 24 hours. DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

[0014] The present disclosure provides compounds and compositions useful for binding to p62, and in some embodiments, such compounds and compositions are useful for inducing degradation of a particular target via binding of p62 and a target of interest. In some embodiments, such compounds include those of the formulae described herein, or a pharmaceutically acceptable salt thereof, wherein each variable is as defined and described herein. Compounds and Definitions

[0015] Compounds of this disclosure include those described generally above and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of Elements, CAS version, Handbook of Chemistry and Physics, 75thEd. Additionally, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry”, 5thEd., Ed.: Smith, M.B. andMarch, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.

[0016] Unless otherwise stated, structures depicted herein are meant to include all stereoisomeric (e.g., enantiomeric or diastereomeric) forms of the structure, as well as all geometric or conformational isomeric forms of the structure. For example, the R and S configurations of each stereocenter are contemplated as part of the disclosure. Therefore, single stereochemical isomers, as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of provided compounds are within the scope of the disclosure. For example, in some cases, Tables 1 and 2 show one or more stereoisomers of a compound, and unless otherwise indicated, represents each stereoisomer alone and / or as a mixture. Unless otherwise stated, all tautomeric forms of provided compounds are within the scope of the disclosure.

[0017] Unless otherwise indicated, structures depicted herein are meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including replacement of hydrogen by deuterium or tritium, or replacement of a carbon by13C- or14C-enriched carbon are within the scope of this disclosure.

[0018] About or approximately: As used herein, the term "approximately" or "about," as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In general, those skilled in the art, familiar within the context, will appreciate the relevant degree of variance encompassed by "about" or "approximately" in that context. For example, in some embodiments, the term "approximately" or "about" may encompass a range of values that are within (i.e., ±) 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.

[0019] Administering: As used herein, the term "administering" or "administration" typically refers to the administration of a composition to a subject to achieve delivery of an agent that is, or is included in, a composition to a target site or a site to be treated. Those of ordinary skill in the art will be aware of a variety of routes that may, in appropriate circumstances, be utilized for administration to a subject, for example a human. For example, in some embodiments, administration may be ocular, oral, parenteral, topical, etc. In some particular embodiments, administration may be bronchial (e.g., by bronchial instillation), buccal, dermal (which may be or comprise, for example, one or more of topical to the dermis, intradermal, interdermal, transdermal,etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a specific organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreal, etc. In some embodiments, administration may be parenteral. In some embodiments, administration may be oral. In some particular embodiments, administration may be intravenous. In some particular embodiments, administration may be subcutaneous. In some embodiments, administration may involve only a single dose. In some embodiments, administration may involve application of a fixed number of doses. In some embodiments, administration may involve dosing that is intermittent (e.g., a plurality of doses separated in time) and / or periodic (e.g., individual doses separated by a common period of time) dosing. In some embodiments, administration may involve continuous dosing (e.g., perfusion) for at least a selected period of time. In some embodiments, administration may comprise a prime- and-boost protocol. A prime-and-boost protocol can include administration of a first dose of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) followed by, after an interval of time, administration of a second or subsequent dose of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine). In the case of an immunogenic composition, a prime-and-boost protocol can result in an increased immune response in a patient.

[0020] Aliphatic: The term “aliphatic” refers to a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as “cycloaliphatic”), that has a single point or more than one points of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-12 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms (e.g., C1-6). In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms (e.g., C1-5). In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms (e.g., C1-4). In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms (e.g., C1-3), and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms (e.g., C1-2). Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl groups and hybrids thereof. A preferred aliphatic group is C1-6alkyl.

[0021] Alkyl: The term “alkyl”, used alone or as part of a larger moiety, refers to a saturated, optionally substituted straight or branched chain hydrocarbon group having (unless otherwise specified) 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2 carbon atoms (e.g., C1-12, C1-10, C1-8, C1-6, C1-4, C1-3, or C1-2). Exemplary alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, and heptyl.

[0022] Alkylene: The term “alkylene” is refers to a bivalent alkyl group. In some embodiments, “alkylene” is a bivalent straight or branched alkyl group. In some embodiments, an "alkylene chain" is a polymethylene group, i.e., -(CH2)n-, wherein n is a positive integer, e.g., from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3. An optionally substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms is optionally replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group and also include those described in the specification herein. It will be appreciated that two substituents of the alkylene group may be taken together to form a ring system. In certain embodiments, two substituents can be taken together to form a 3- to 7-membered ring. The substituents can be on the same or different atoms. The suffix “-ene” or “-enyl” when appended to certain groups herein are intended to refer to a bifunctional moiety of said group. For example, “-ene” or “-enyl”, when appended to “cyclopropyl” becomes “cyclopropylene” or “cyclopropylenyl” and is intended to refer to a bifunctional cyclopropyl group, e.g.,.

[0023] Alkenyl: The term “alkenyl”, used alone or as part of a larger moiety, refers to an optionally substituted straight or branched chain or cyclic hydrocarbon group having at least one double bond and having (unless otherwise specified) 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3 carbon atoms(e.g., C2-12, C2-10, C2-8, C2-6, C2-4, or C2-3). Exemplary alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, and heptenyl. The term “cycloalkenyl” refers to an optionally substituted non-aromatic monocyclic or multicyclic ring system containing at least one carbon-carbon double bond and having about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, and cycloheptenyl.

[0024] Alkynyl: The term “alkynyl”, used alone or as part of a larger moiety, refers to an optionally substituted straight or branched chain hydrocarbon group having at least one triple bond and having (unless otherwise specified) 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3 carbon atoms (e.g., C2-12, C2-10, C2-8, C2-6, C2-4, or C2-3). Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and heptynyl.

[0025] Antagonist: As will be understood by those skilled in the art, the term “antagonist” generally refers to an agent whose presence or level correlates with decreased level or activity of a target, as compared with that observed absent the agent (or with the agent at a different level). In some embodiments, an antagonist is one whose presence or level correlates with a target level or activity that is comparable to or less than a particular reference level or activity (e.g., that observed under appropriate reference conditions, such as presence of a known antagonist, e.g., a positive control). In some embodiments, an antagonist may be a direct antagonist in that it exerts its influence directly on (e.g., interacts directly with) the target; in some embodiments, an antagonist may be an indirect antagonist in that it exerts its influence indirectly (e.g., by acting on, such as interacting with, a regulator of the target, or with some other component or entity.

[0026] Aryl: The term “aryl” refers to monocyclic and bicyclic ring systems having a total of six to fourteen ring members (e.g., C6-C14), wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. In some embodiments, an “aryl” group contains between six and twelve total ring members (e.g., C6-C12). The term “aryl” may be used interchangeably with the term “aryl ring”. In certain embodiments, “aryl” refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Unless otherwise specified, “aryl” groups are hydrocarbons. In some embodiments, an “aryl” ring system is an aromatic ring (e.g., phenyl) that is fused to a non-aromatic ring (e.g., cycloalkyl). Examples of aryl rings include that are fused include.

[0027] Bicyclic: The term “bicyclic ring” or “bicyclic ring system” refers to any bicyclic ring system, i.e., carbocyclic or heterocyclic, saturated or having one or more units of unsaturation, having one or more atoms in common between the two rings of the ring system. Thus, the term includes any permissible ring fusion, such as ortho-fused or spirocyclic. As used herein, the term “heterobicyclic” is a subset of “bicyclic” that requires that one or more heteroatoms are present in one or both rings of the bicycle. Such heteroatoms may be present at ring junctions and are optionally substituted, and may be selected from nitrogen (including N-oxides), oxygen, sulfur (including oxidized forms such as sulfones and sulfonates), phosphorus (including oxidized forms such as phosphates), boron, etc. In some embodiments, a bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. As used herein, theterm “bridged bicyclic” refers to any bicyclic ring system, i.e., carbocyclic or heterocyclic, saturated or partially unsaturated, having at least one bridge. As defined by IUPAC, a “bridge” is an unbranched chain of atoms or an atom or a valence bond connecting two bridgeheads, where a “bridgehead” is any skeletal atom of the ring system which is bonded to three or more skeletal atoms (excluding hydrogen). In some embodiments, a bridged bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Such bridged bicyclic groups are well known in the art and include those groups set forth below where each group is attached to the rest of the molecule at any substitutable carbon or nitrogen atom. Unless otherwise specified, a bridged bicyclic group is optionally substituted with one or more substituents as set forth for aliphatic groups. Additionally or alternatively, any substitutable nitrogen of a bridged bicyclic group is optionally substituted. Exemplary bicyclic rings include:Exemplary bridged bicyclics include:

[0028] Biological sample: As used herein, the term “biological sample” typically refers to a sample obtained or derived from a biological source (e.g., a tissue or organism or cell culture) of interest, as described herein. In some embodiments, a source of interest comprises an organism, such as an animal or human. In some embodiments, a biological sample is or comprises biological tissue or fluid. In some embodiments, a biological sample may be or comprise bone marrow; blood; blood cells; ascites; tissue or fine needle biopsy samples; cell-containing body fluids; free floating nucleic acids; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph; gynecological fluids; skin swabs; vaginal swabs; oral swabs; nasal swabs; washings or lavages such as a ductal lavages or broncheoalveolar lavages; aspirates; scrapings; bone marrow specimens; tissue biopsy specimens; surgical specimens; feces, other body fluids, secretions, and / or excretions; and / or cells therefrom, etc. In some embodiments, a biological sample is or comprises cells obtained from an individual. In some embodiments, obtained cells are or include cells from an individual from whom the sample is obtained. In some embodiments, a sample is a “primaiy sample” obtained directly from a source of interest by any appropriate means. For example, in some embodiments, a primaiy biological sample is obtained by methods selected from the group consisting of biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of body fluid (e g., blood, lymph, feces etc.), etc. In some embodiments, as will be clear from context, the term “sample” refers to a preparation that is obtained by processing (e.g., by removing one or more components of and / or by adding one or more agents to) a primary sample. For example, filtering using a semi-permeable membrane. Such a “processed sample” may comprise, for example, nucleic acids or proteins extracted from a sample or obtained by subjecting a primarysample to techniques such as amplification or reverse transcription of mRNA, isolation and / or purification of certain components, etc.

[0029] Carrier: As used herein, the term “carrier” refers to a diluent, adjuvant, excipient, or vehicle with which a composition is administered. In some exemplary embodiments, carriers can include sterile liquids, such as, for example, water and oils, including oils of petroleum, animal, vegetable or synthetic origin, such as, for example, peanut oil, soybean oil, mineral oil, sesame oil and the like. In some embodiments, carriers are or include one or more solid components.

[0030] Composition: Those skilled in the art will appreciate that the term “composition” may be used to refer to a discrete physical entity that comprises one or more specified components. In general, unless otherwise specified, a composition may be of any form – e.g., gas, gel, liquid, solid, etc.

[0031] Cycloaliphatic: As used herein, the term “cycloaliphatic” refers to a monocyclic C3-8hydrocarbon or a bicyclic C6-12hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point or more than one points of attachment to the rest of the molecule.

[0032] Cycloalkyl: As used herein, the term “cycloalkyl” refers to an optionally substituted saturated ring monocyclic or polycyclic system of about 3 to about 10 ring carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0033] Dosage form or unit dosage form: Those skilled in the art will appreciate that the term “dosage form” may be used to refer to a physically discrete unit of an active agent (e.g., a therapeutic or diagnostic agent) for administration to a subject. Typically, each such unit contains a predetermined quantity of active agent. In some embodiments, such quantity is a unit dosage amount (or a whole fraction thereof) appropriate for administration in accordance with a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., with a therapeutic dosing regimen).

[0034] Dosing regimen or therapeutic regimen: Those skilled in the art will appreciate that the terms “dosing regimen” and “therapeutic regimen” may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses. In some embodiments, a dosing regimencomprises a plurality of doses each of which is separated in time from other doses. In some embodiments, individual doses are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen).

[0035] Effective Amount: The term “effective amount” refers to the amount of a compound sufficient to effect beneficial or desired results (e.g., a therapeutic, ameliorative, inhibitory, or preventative result). An effective amount can be administered in one or more administrations, applications, or dosages and is not intended to be limited to a particular formulation or administration route.

[0036] Excipient: As used herein, the term “excipient” refers to a non-therapeutic agent that may be included in a pharmaceutical composition, for example, to provide or contribute to a desired consistency or stabilizing effect. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like.

[0037] Halogen: The term “halogen” or “halo” means F, Cl, Br, or I.

[0038] Heteroaliphatic: The term “heteroaliphatic” or “heteroaliphatic group”, as used herein, denotes an optionally substituted hydrocarbon moiety having, in addition to carbon atoms, from one to five heteroatoms, that may be straight–chain (i.e., unbranched), branched, or cyclic (“heterocyclic”) and may be completely saturated or may contain one or more units of unsaturation, but which is not aromatic. The term “heteroatom” refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen. The term “nitrogen” also includes a substituted nitrogen. Unless otherwise specified, heteroaliphatic groups contain 1–10 carbon atoms wherein 1–3 carbon atoms are optionally andindependently replaced with heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, heteroaliphatic groups contain 1–4 carbon atoms, wherein 1–2 carbon atoms are optionally and independently replaced with heteroatoms selected from oxygen, nitrogen, and sulfur. In yet other embodiments, heteroaliphatic groups contain 1–3 carbon atoms, wherein 1 carbon atom is optionally and independently replaced with a heteroatom selected from oxygen, nitrogen, and sulfur. Suitable heteroaliphatic groups include, but are not limited to, linear or branched, heteroalkyl, heteroalkenyl, and heteroalkynyl groups. For example, a 1- to 10 atom heteroaliphatic group includes the following exemplary groups: -O-CH3, -CH2-O-CH3, -O-CH2- CH2-O-CH2-CH2-O-CH3, and the like.

[0039] Heteroaryl: The terms “heteroaryl” and “heteroar–”, used alone or as part of a larger moiety, e.g., “heteroaralkyl”, or “heteroaralkoxy”, refer to monocyclic or bicyclic ring groups having 5 to 10 ring atoms (e.g., 5- to 6-membered monocyclic heteroaryl or 9- to 10-membered bicyclic heteroaryl); having 6, 10, or 14 π-electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl, imidazo[1,2-a]pyrimidinyl, imidazo[1,2-a]pyridyl, imidazo[4,5-b]pyridyl, imidazo[4,5-c]pyridyl, pyrrolopyridyl, pyrrolopyrazinyl, thienopyrimidinyl, triazolopyridyl, and benzoisoxazolyl. The terms “heteroaryl” and “heteroar–”, as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring (i.e., a bicyclic heteroaryl ring having 1 to 3 heteroatoms). Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzotriazolyl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H–quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyrido[2,3–b]–1,4–oxazin–3(4H)–one, 4H-thieno[3,2-b]pyrrole, and benzoisoxazolyl. The term “heteroaryl” may be used interchangeably with the terms “heteroaryl ring”, “heteroaryl group”, or “heteroaromatic”, any of which terms include rings that are optionally substituted. Example monocyclic and bicyclic heteroaryl rings include:

[0040] Heteroatom: The term “heteroatom” as used herein refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen.

[0041] Heterocycle: As used herein, the terms “heterocycle”, “heterocyclyl”, “heterocyclic radical”, and “heterocyclic ring” are used interchangeably and refer to a stable 3- to 8-membered monocyclic, a 6- to 10-membered bicyclic, or a 10- to 16-membered polycyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, such as one to four, heteroatoms, as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes a substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0–3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR+(as in N-substituted pyrrolidinyl). A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and thiamorpholinyl. A heterocyclyl group may be mono-, bi-, tri-, or polycyclic, preferably mono-, bi-, or tricyclic, more preferably mono- or bicyclic. A bicyclic heterocyclic ring also includes groups in which the heterocyclic ring is fused to one or more aryl rings. Exemplary bicyclic heterocyclic groups include indolinyl, isoindolinyl, benzodioxolyl, 1,3-dihydroisobenzofuranyl, 2,3-dihydrobenzofuranyl, and tetrahydroquinolinyl. A bicyclic heterocyclic ring can also be a spirocyclic ring system (e.g., 7- to 11-membered spirocyclic fused heterocyclic ring having, in addition to carbon atoms, one or more heteroatoms as defined above (e.g., one, two, three or four heteroatoms)). A bicyclic heterocyclic ring can also be a bridged ring system (e.g., 7- to 11-membered bridged heterocyclic ring having one, two, or three bridging atoms.

[0042] Modulator: The term “modulator,” as used herein, refers to a compound (e.g., a small molecule) that can alter the activity of another molecule (e.g., a protein). For example, in someembodiments, a modulator can cause an increase or decrease in the magnitude of a certain activity of a type of molecule as compared to the magnitude of the activity in the absence of the modulator. For example, a modulator can be an agonist or an antagonist of a particular target, as those terms are defined herein. For example, in some embodiments, a modulator is an agonist. In some embodiments, a modulator is an antagonist.

[0043] Oral: The phrases “oral administration” and “administered orally” as used herein have their art-understood meaning referring to administration by mouth of a compound or composition.

[0044] Parenteral: The phrases “parenteral administration” and “administered parenterally” as used herein have their art-understood meaning referring to modes of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0045] Partially unsaturated: As used herein, the term “partially unsaturated” refers to a ring moiety that includes at least one double or triple bond between ring atoms. The term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic (e.g., aryl or heteroaryl) moieties, as herein defined.

[0046] Patient or subject: As used herein, the term “patient” or “subject” refers to any organism to which a provided composition is or may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and / or therapeutic purposes. Typical patients or subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, a patient is a human. In some embodiments, a patient or a subject is suffering from or susceptible to one or more disorders or conditions. In some embodiments, a patient or subject displays one or more symptoms of a disorder or condition. In some embodiments, a patient or subject has been diagnosed with one or more disorders or conditions. In some embodiments, a patient or a subject is receiving or has received certain therapy to diagnose and / or to treat a disease, disorder, or condition.

[0047] Pharmaceutical composition: As used herein, the term “pharmaceutical composition” refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in unit dose amount appropriate for administration in a therapeutic or dosing regimen that shows a statistically significant probabilityof achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream, or foam; sublingually; ocularly; transdermally; or nasally, pulmonary, and to other mucosal surfaces.

[0048] Pharmaceutically acceptable: As used herein, the phrase “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0049] Pharmaceutically acceptable salt: The term “pharmaceutically acceptable salt”, as used herein, refers to salts of such compounds that are appropriate for use in pharmaceutical contexts, i.e., salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2–hydroxy–ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2–naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3–phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p–toluenesulfonate, undecanoate, valerate salts, and the like.

[0050] Further, acids which are generally considered suitable for the formation of pharmaceutically useful salts from basic pharmaceutical compounds are discussed, for example, by P. Stahl et al., Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley-VCH; S. Berge et al., Journal of Pharmaceutical Sciences 1977, 66(1), 1-19; P. Gould, International J. of Pharmaceutics 1986, 33, 201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; and in The Orange Book (Food & Drug Administration, Washington, D.C. on their website). These disclosures are incorporated herein by reference.

[0051] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(C1–4alkyl)4salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.

[0052] Polypeptide: The term “polypeptide”, as used herein, generally has its art-recognized meaning of a polymer of at least three amino acids, e.g., linked to each other by peptide bonds. Those of ordinary skill in the art will appreciate that the term “polypeptide” is intended to be sufficiently general as to encompass not only polypeptides having a complete sequence recited herein, but also to encompass polypeptides that represent functional fragments (i.e., fragments retaining at least one activity) of such complete polypeptides. Moreover, those of ordinary skill in the art understand that protein sequences generally tolerate some substitution without destroying activity. Thus, any polypeptide that retains activity and shares at least about 30-40% overall sequence identity, often greater than about 50%, 60%, 70%, or 80%, and further usually including at least one region of much higher identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99% in one or more highly conserved regions, usually encompassing at least 3-4 and often up to 20 or more amino acids, with another polypeptide of the same class, is encompassed within the relevant term “polypeptide” as used herein. Polypeptides may contain L-amino acids, D-aminoacids, or both and may contain any of a variety of amino acid modifications or analogs known in the art. Useful modifications include, e.g., terminal acetylation, amidation, methylation, etc. In some embodiments, proteins may comprise natural amino acids, non-natural amino acids, synthetic amino acids, and combinations thereof.

[0053] Prevent or prevention: As used herein, the terms “prevent” or “prevention”, when used in connection with the occurrence of a disease, disorder, and / or condition, refer to reducing the risk of developing the disease, disorder and / or condition and / or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time.

[0054] Substituted or optionally substituted: As described herein, compounds of the invention may contain “optionally substituted” moieties. In general, the term “substituted,” whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. “Substituted” applies to one or more hydrogens that are either explicit or implicit from the structure (e.g.,refers to at leastrefers to at least). Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds. The term “stable,” as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes provided herein. Groups described as being “substituted” preferably have between 1 and 4 substituents, more preferably 1 or 2 substituents. Groups described as being “optionally substituted” may be unsubstituted or be “substituted” as described above.

[0055] Suitable monovalent substituents on a substitutable carbon atom of an “optionally substituted” group are independently halogen; –(CH2)0–4R°; –(CH2)0–4OR°; -O(CH2)0–4R°, –O– (CH2)0–4C(O)OR°; –(CH2)0–4CH(OR°)2; –(CH2)0–4SR°; –(CH2)0–4Ph, which may be substituted with R°; –(CH2)0–4O(CH2)0–1Ph which may be substituted with R°; –CH=CHPh, which may be substituted with R°; –(CH2)0–4O(CH2)0–1-pyridyl which may be substituted with R°; –NO2; –CN; –N3; -(CH2)0–4N(R°)2; –(CH2)0–4N(R°)C(O)R°; –N(R°)C(S)R°; –(CH2)0–4N(R°)C(O)NR°2; -N(R°)C(S)NR°2; –(CH2)0–4N(R°)C(O)OR°; - N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)NR°2; -N(R°)N(R°)C(O)OR°; –(CH2)0–4C(O)R°; C(S)R°; –(CH2)0–4C(O)OR°; –(CH2)0–4C(O)SR°; -(CH2)0–4C(O)OSiR°3; –(CH2)0–4OC(O)R°; – OC(O)(CH2)0–4SR°; –(CH2)0–4SC(O)R°; –(CH2)0–4C(O)NR°2; –C(S)NR°2; –C(S)SR°; – SC(S)SR°, -(CH2)0–4OC(O)NR°2; -C(O)N(OR°)R°; –C(O)C(O)R°; –C(O)CH2C(O)R°; – C(NOR°)R°; -(CH2)0–4SSR°; –(CH2)0–4S(O)2R°; –(CH2)0–4S(O)2OR°; –(CH2)0–4OS(O)2R°; – S(O)2NR°2; -(CH2)0–4S(O)R°; -N(R°)S(O)2NR°2; –N(R°)S(O)2R°; –N(OR°)R°; –C(NH)NR°2; – P(O)2R°; -P(O)R°2; -OP(O)R°2; –OP(O)(OR°)2; SiR°3; –(C1–4straight or branched alkylene)O– N(R°)2; or –(C1–4straight or branched alkylene)C(O)O–N(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, C1–6aliphatic, –CH2Ph, –O(CH2)0–1Ph, -CH2-(5- to 6-membered heteroaryl ring), or a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R°, taken together with their intervening atom(s), form a 3- to 12-membered saturated, partially unsaturated, or aryl mono– or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.

[0056] Suitable monovalent substituents on R° (or the ring formed by taking two independent occurrences of R° together with their intervening atoms), are independently halogen, –(CH2)0–2R●, –(haloR●), –(CH2)0–2OH, –(CH2)0–2OR●, –(CH2)0–2CH(OR●)2, -O(haloR●), –CN, –N3, –(CH2)0–2C(O)R●, –(CH2)0–2C(O)OH, –(CH2)0–2C(O)OR●, –(CH2)0–2SR●, –(CH2)0–2SH, –(CH2)0–2NH2, – (CH2)0–2NHR●, –(CH2)0–2NR●2, –NO2, –SiR●3, –OSiR●3, -C(O)SR●, –(C1–4straight or branched alkylene)C(O)OR●, or –SSR●wherein each R●is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently selected from C1–4aliphatic, – CH2Ph, –O(CH2)0–1Ph, or a 3- to 6-membered saturated, partially unsaturated, or aryl ring having0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =O and =S.

[0057] Suitable divalent substituents on a saturated carbon atom of an “optionally substituted” group include the following: =O (“oxo”), =S, =NNR*2, =NNHC(O)R*, =NNHC(O)OR*, =NNHS(O)2R*, =NR*, =NOR*, –O(C(R*2))2–3O–, or –S(C(R*2))2–3S–, wherein each independent occurrence of R*is selected from hydrogen, C1–6aliphatic which may be substituted as defined below, or an unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0–4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that are bound to vicinal substitutable carbons of an “optionally substituted” group include: –O(CR*2)2–3O–, wherein each independent occurrence of R*is selected from hydrogen, C1–6aliphatic which may be substituted as defined below, or an unsubstituted 5–6–membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0058] Suitable substituents on the aliphatic group of R*include halogen, –R●, -(haloR●), -OH, –OR●, –O(haloR●), –CN, –C(O)OH, –C(O)OR●, –NH2, –NHR●, –NR●2, or –NO2, wherein each R●is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C1–4aliphatic, –CH2Ph, –O(CH2)0–1Ph, or a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0059] Suitable substituents on a substitutable nitrogen of an “optionally substituted” group include –R†, –NR†2, –C(O)R†, –C(O)OR†, –C(O)C(O)R†, – C(O)CH2C(O)R†, -S(O)2R†, -S(O)2NR†2, –C(S)NR†2, –C(NH)NR†2, or –N(R†)S(O)2R†; wherein each R†is independently hydrogen, C1–6aliphatic which may be substituted as defined below, unsubstituted –OPh, or an unsubstituted 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R†, taken together with their intervening atom(s) form an unsubstituted 3- to 12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0060] Suitable substituents on the aliphatic group of R†are independently halogen, – R●, -(haloR●), –OH, –OR●, –O(haloR●), –CN, –C(O)OH, –C(O)OR●, –NH2, –NHR●, –NR●2,or -NO2, wherein each R●is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C1–4aliphatic, –CH2Ph, –O(CH2)0–1Ph, or a 3- to 6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0061] Small molecule: As used herein, the term “small molecule” means a low molecular weight organic and / or inorganic compound. In general, a “small molecule” is a molecule that is less than about 5 kilodaltons (kD) in size. In some embodiments, a small molecule is less than about 4 kD, 3 kD, about 2 kD, or about 1 kD. In some embodiments, the small molecule is less than about 800 daltons (D), about 600 D, about 500 D, about 400 D, about 300 D, about 200 D, or about 100 D. In some embodiments, a small molecule is less than about 2000 g / mol, less than about 1500 g / mol, less than about 1000 g / mol, less than about 800 g / mol, or less than about 500 g / mol. In some embodiments, a small molecule is not a polymer.

[0062] In some embodiments, a small molecule does not include a polymeric moiety. In some embodiments, a small molecule is not and / or does not comprise a protein or polypeptide (e.g., is not an oligopeptide or peptide). In some embodiments, a small molecule is not and / or does not comprise a polynucleotide (e.g., is not an oligonucleotide). In some embodiments, a small molecule is not and / or does not comprise a polysaccharide; for example, in some embodiments, a small molecule is not a glycoprotein, proteoglycan, glycolipid, etc.). In some embodiments, a small molecule is not a lipid.

[0063] In some embodiments, a small molecule is a modulating agent (e.g., is an inhibiting agent or an activating agent). In some embodiments, a small molecule is biologically active. In some embodiments, a small molecule is detectable (e.g., comprises at least one detectable moiety). In some embodiments, a small molecule is a therapeutic agent.

[0064] Those of ordinary skill in the art, reading the present disclosure, will appreciate that certain small molecule compounds described herein may be provided and / or utilized in any of a variety of forms such as, for example, crystal forms (e.g., polymorphs, solvates, etc), salt forms, protected forms, pro-drug forms, ester forms, isomeric forms (e.g., optical and / or structural isomers), isotopic forms, etc.

[0065] Those of ordinary skill in the art will appreciate that certain small molecule compounds have structures that can exist in one or more steroisomeric forms. In some embodiments, such a small molecule may be utilized in accordance with the present disclosure in the form of anindividual enantiomer, diastereomer or geometric isomer, or may be in the form of a mixture of stereoisomers; in some embodiments, such a small molecule may be utilized in accordance with the present disclosure in a racemic mixture form.

[0066] Those of skill in the art will appreciate that certain small molecule compounds have structures that can exist in one or more tautomeric forms. In some embodiments, such a small molecule may be utilized in accordance with the present disclosure in the form of an individual tautomer, or in a form that interconverts between tautomeric forms.

[0067] Those of skill in the art will appreciate that certain small molecule compounds have structures that permit isotopic substitution (e.g.,2H or3H for H;11C,13C or14C for12C;13N or15N for14N;17O or18O for16O;36Cl for35Cl or37Cl;18F for19F;131I for127I; etc.). In some embodiments, such a small molecule may be utilized in accordance with the present disclosure in one or more isotopically modified forms, or mixtures thereof.

[0068] In some embodiments, reference to a particular small molecule compound may relate to a specific form of that compound. In some embodiments, a particular small molecule compound may be provided and / or utilized in a salt form (e.g., in an acid-addition or base-addition salt form, depending on the compound); in some such embodiments, the salt form may be a pharmaceutically acceptable salt form.

[0069] In some embodiments, where a small molecule compound is one that exists or is found in nature, that compound may be provided and / or utilized in accordance in the present disclosure in a form different from that in which it exists or is found in nature. Those of ordinary skill in the art will appreciate that, in some embodiments, a preparation of a particular small molecule compound that contains an absolute or relative amount of the compound, or of a particular form thereof, that is different from the absolute or relative (with respect to another component of the preparation including, for example, another form of the compound) amount of the compound or form that is present in a reference preparation of interest (e.g., in a primary sample from a source of interest such as a biological or environmental source) is distinct from the compound as it exists in the reference preparation or source. Thus, in some embodiments, for example, a preparation of a single stereoisomer of a small molecule compound may be considered to be a different form of the compound than a racemic mixture of the compound; a particular salt of a small molecule compound may be considered to be a different form from another salt form of the compound; a preparation that contains only a form of the compound that contains one conformational isomer((Z) or (E)) of a double bond may be considered to be a different form of the compound from one that contains the other conformational isomer ((E) or (Z)) of the double bond; a preparation in which one or more atoms is a different isotope than is present in a reference preparation may be considered to be a different form; etc.

[0070] Those skilled in the art will appreciate that a bond designated asin a small molecule structure, as used herein, refers to a bond that, in some embodiments, is a single (e.g., saturated) bond, and in some embodiments, is a double (e.g., unsaturated) bond. For example, the following structure:is intended to encompass both.

[0071] Those skilled in the art will further appreciate that, in small molecule structures, the symbols and ---, as used herein, are interchangeable, and each refer to a point of attachment between two atoms. Additionally or alternatively, the symbolrefers to a point of attachment ring in a spirocyclic manner.

[0072] Treat: As used herein, the terms “treat,” “treatment,” or “treating” refer to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and / or condition, for example, for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and / or condition. Compounds for Binding p62

[0073] In certain aspects, the present disclosure provides compounds for binding to p62. In some embodiments, said compounds are bifunctional, where they comprise a p62 binding moiety, that binds or associates with p62, and a target binding moiety, that binds or associates with a target, and thereby promotes autophagic degradation of the target. As used herein, a moiety “associates”with a target or p62 through various chemical interactions known to those of skill in the art. For example, a moiety can associate with a target or p62 by hydrogen bonding, Van der Waals forces, London dispersion forces, ionic bonding, and the like. In some embodiments, p62 binding is determined according to methods described herein, for example in Example C1. In some embodiments, a compound for binding p62 and a target of interest is a compound of formula I: A-B-C I or a pharmaceutically acceptable salt thereof, wherein A is a moiety that binds or associates with p62; B is a linker moiety; and C is a target binding moiety.

[0074] In some embodiments, A is a polypeptide or peptidomimetic moiety that binds or associates with p62. In some embodiments, A is a moiety comprising two or three conjugated amino acids (e.g., a canonical or non-canonical amino acid) linked by a peptide bond (an amide linking two consecutive amino acids from C1 of one amino acid to N2 of another amino acid). In some embodiments, A is a peptidomimetic moiety. A peptidomimetic moiety, as used herein, is a small protein-like chain that mimics a peptide. In some embodiments, a peptidomimetic is a moiety that comprises modified peptides, structural mimetics to peptides, peptidic foldamers, mechanistic mimetics, or other structural analogs.

[0075] In some embodiments, A is a moiety:and a compound of formula I is a compound of formula II-1 or II-2:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)- C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2,wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, - C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety.

[0076] In some embodiments, a compound described herein is of formula II-1:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)- C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, - C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety.

[0077] In some embodiments, a compound described herein is of formula II-2:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)- C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, - C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety.

[0078] In some embodiments, A is a moiety:and a compound of formula I is a compound of formula III-1 or III-2:or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)- C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, - C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety.

[0079] In some embodiments, a compound described herein is a compound of formula III-1:or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)- C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc;each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, - C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety.

[0080] In some embodiments, a compound described herein is a compound of formula III-2:or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)- C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen;each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, - C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety.

[0081] Descriptions of values and variables herein are intended to apply to all provided formulae (e.g., any of formula II-1, II-2, III-1, III-2, etc.), unless otherwise specified.

[0082] As described generally herein, Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S. In some embodiments, Ring A is triazole, imidazole, oxazole, thiazole, pyrrole, pyrazole, pyridine, pyrimidine, pyrazine, or triazine.

[0083] In some embodiments, Ring A is selected from:where --- represents a point o3f attachment to G , andrepresents a point of attachment to G2.

[0084] As described generally herein, G1is 5- to 12-membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb. In some embodiments, G1is 5- to 12- membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, C6-C12aryl or guanidine, wherein G1is optionally substituted with one or more Rb. In some embodiments, G1is 5- to 12- membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S or 4- to 6- membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, optionally substituted with one or more Rb.

[0085] In some embodiments, G1is 5- to 12-membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, and optionally substituted with one or more Rb. In someembodiments, G1is 5- to 6-membered monocyclic heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S, and optionally substituted with one or more Rb. In some embodiments, G1is unsubstituted 5- to 6-membered monocyclic heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S. In some embodiments, G1is 5- to 6-membered monocyclic heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S, and substituted with one or more Rb. In some embodiments, G1is 5- to 6-membered monocyclic heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S, substituted with one or more of halogen, -NH2, or optionally substituted C1-C6aliphatic. In some embodiments, G1is 5- to 6-membered monocyclic heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S, substituted with one or more of halogen, -NH2, or C1-C6aliphatic substituted with –C(NH)NR°2or –(CH2)0–4C(O)NR°2. In some embodiments, G1is 5- to 6-membered monocyclic heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S, substituted with one or more of -NH2or -CH3. In some embodiments, G1is imidazolyl, pyridinyl, pyrrolyl, pyrazolyl, pyrimidinyl, or thiazolyl substituted with one or more of Rb. In some embodiments, G1is imidazolyl, pyridinyl, pyrrolyl, pyrazolyl, pyrimidinyl, or thiazolyl, substituted with one or more of halogen, -NH2, or C1-C6aliphatic. In some embodiments, G1is optionally substituted imidazolyl or pyridinyl substituted with one or more of halogen, -NH2,or C1-C6aliphatic. In some embodiments, G1is unsubstituted imidazolyl or pyridinyl. In some embodiments, G1is imidazolyl or pyridinyl substituted with one or more of –NH2or -CH3.

[0086] In some embodiments, G1is 7- to 12-membered bicyclic heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, optionally substituted with one or more Rb. In some embodiments, G1is 7 to 12-membered bicyclic heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, optionally substituted with one or more –N(Ra)2or halogen. In some embodiments, G1is 7- to 12-membered bicyclic heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, substituted with one or more of halogen, -NH2, or optionally substituted C1-C6aliphatic. In some embodiments, G1is 7- to 12-membered bicyclic heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, optionally substituted with one or more of halogen, -NH2,or –CH3. In some embodiments, G1is benzoimidazolyl, indazolyl, indolyl, or pyrrolopyridinyl optionally substituted with one or more Rb. In some embodiments, G1is benzoimidazolyl or indolyl optionally substituted with one or more Rb. In some embodiments, G1is benzoimidazolyl, indazolyl, indolyl, or pyrrolopyridinyl substituted with one or more of halogen, -NH2,or C1-C6aliphatic. In some embodiments, G1is benzoimidazolyl or indolyl substituted with one or more ofhalogen, -NH2,or C1-C6aliphatic. In some embodiments, G1is benzoimidazolyl, indazolyl, indolyl, or pyrrolopyridinyl substituted with one or more of -NH2or CH3. In some embodiments, G1is optionally substituted benzoimidazolyl or indolyl substituted with one or more of –NH2or – CH3.

[0087] In some embodiments, G1is 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S and optionally substituted with one or more Rb. In some embodiments, G1is 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S and optionally substituted with one or more of halogen, -NH2, or C1-C6aliphatic. In some embodiments, G1is 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S and optionally substituted with -NH2or –CH3.

[0088] In some embodiments, G1is C6-C12aryl optionally substituted with one or more Rb. In some embodiments, G1is C6-C12aryl optionally substituted with one or more of halogen, -N(R°)2, R°, -NO2, or –C(NH)N(R°)2. In some embodiments, G1is C6-C12aryl optionally substituted with halogen or –NH2.

[0089] In some embodiments, G1is guanidine.

[0090] In some embodiments, G1is –C(O)NH2.

[0091] In some embodiments, G1is selected from:

[0092] In some embodiments, G1is selected from:wherein represents a point of attachment to moiety B, andrepresents a point of attachment to moiety G2.

[0093] In some embodiments, G1is selected from:whereinrepresents a point of attachment to moiety B, and2represents a point of attachment to moiety G .

[0094] As described generally herein, G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, or C1-C6aliphatic-4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, wherein G2is optionally substituted with one or more Rb.

[0095] In some embodiments, G2is C1-C6aliphatic, optionally substituted with one or more Rb. In some embodiments, G2is C1-C6alkyl, optionally substituted with one or more Rb. In some embodiments, G2is C1-C6alkyl, optionally substituted with –N(Ra)2. In some embodiments, G2is C1-C6alkyl, substituted with –N(Ra)2. In some embodiments, G2is methylene, ethylene, propylene or n-butlyene, substituted with –NH2.

[0096] In some embodiments, G2is –N(Ra)-C1-C6aliphatic, optionally substituted with one or more Rb. In some embodiments, G2is –NH-C1-C6aliphatic, optionally substituted with one or more Rb. In some embodiments, G2is –NH-C1-C6alkyl, optionally substituted with one or more Rb. In some embodiments, G2is –NH-C1-C6alkylene, optionally substituted with C1-C6aliphatic.

[0097] In some embodiments, G2is C1-C6aliphatic–N(Ra)-, optionally substituted with one or more Rb. In some embodiments, G2is C1-C6aliphatic-NH-, optionally substituted with one ormore Rb. In some embodiments, G2is C1-C6alkylene-NH-, optionally substituted with one or more Rb. In some embodiments, G2is C1-C6alkylene-NH-, optionally substituted with C1-C6aliphatic.

[0098] In some embodiments, G2is –O-C1-C6aliphatic, optionally substituted with one or more Rb. In some embodiments, G2is –O-C1-C6alkylene, optionally substituted with one or more Rb.

[0099] In some embodiments, G2is –C(O)-C1-C6aliphatic, optionally substituted with one or more Rb. In some embodiments, G2is –C(O)-C1-C6alkylene, optionally substituted with one or more Rb.

[0100] In some embodiments, G2is C6-C12aryl-C0-C6aliphatic, optionally substituted with one or more Rb. In some embodiments, G2is C6-C12aryl, optionally substituted with one or more Rb. In some embodiments, G2is phenyl, optionally substituted with one or more Rb. In some embodiments, G2is C6-C12aryl-C1-C6aliphatic, optionally substituted with one or more Rb. In some embodiments, G2is phenyl-C1-C6aliphatic, optionally substituted with one or more Rb.

[0101] In some embodiments, G2is (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, optionally substituted with one or more Rb. In some embodiments, G2is (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, optionally substituted with one or more –N(Ra)2. In some embodiments, G2is piperdine-C1-C6aliphatic, optionally substituted with one or more N(Ra)2. In some embodiments, G2is piperdine-C1-C6aliphatic, optionally substituted with one or more NH2.

[0102] In some embodiments, G2is 2- to 10-membered heteroaliphatic, optionally substituted with one or more Rb. In some embodiments, G2–C1-C3aliphatic-NH-C1-C3aliphatic, optionally substituted with one or more Rb.

[0103] In some embodiments, G2is selected from:whereinrepresents a point of attachment to –C(O)-N(Ra)- in formulae II-1 or II-2, or to A in formulae III-1 or III-2, andrepresents a point of attachment to G1.

[0104] As described herein, G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc. In some embodiments (e.g., of compounds of formula II-1 or III-1), G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc. In some embodiments (e.g., of compounds of formula II-2 or III- 2), G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C1-C6aliphatic, C3-C12cycloaliphatic, 4- to9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C6aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, wherein G3is optionally substituted with one or more Rc.

[0105] In some embodiments, G3is C1-C7aliphatic, optionally substituted with one or more Rc. In some embodiments, G3is C1-C7alkyl, optionally substituted with one or more Rc. In some embodiments, G3is methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, hexyl or heptyl. In some embodiments, G3is C1-C7alkylene, optionally substituted with one or more Rc. In some embodiments, G3is methylene, ethylene, propylene, butylene, pentylene, hexylene, or heptylene. In some embodiments, G3is C1-C7aliphatic, optionally substituted with one or more of: optionally substituted C3-C6cycloaliphatic, -ORa, or -C(O)-ORa. In some embodiments, G3is C1-C7aliphatic optionally substituted with an optionally substituted C3-C6cycloaliphatic. In some embodiments, G3is C1-C7aliphatic optionally substituted with an optionally substituted C1-C6aliphatic. In some embodiments, G3is C1-C7aliphatic optionally substituted with C1-C6aliphatic, which is optionally substituted with –(CH2)0–4OR° or –(CH2)0–4SR°. In some embodiments, G3is C1-C6aliphatic optionally substituted with -ORa.

[0106] In some embodiments, G3is C1-C6aliphatic-C(O)-N(Ra)-C0-C6aliphatic, optionally substituted with one or more Rc. In some embodiments, G3is C1-C6aliphatic-C(O)-N(Ra)-, optionally substituted with one or more Rc. In some embodiments, G3is C1-C6aliphatic-C(O)- N(Ra)-C1-C6aliphatic, optionally substituted with one or more Rc. In some embodiments, G3is C1-C6aliphatic-C(O)-N(Ra)-C1-C6aliphatic, optionally substituted with one or more -ORa.

[0107] In some embodiments of formula II-1 or III-1, G3is:wherein --- represents a point of attachment to moiety B, andrepresents a point of attachment to –C(O)-N(Ra) in formula II-1, and Ring A in formula III-1.

[0108] In some embodiments of formula II-2 or III-2, G3is:.

[0109] In some embodiments, G3is C3-C12cycloaliphatic optionally substituted with one or more Rc. In some embodiments, G3is C3-C6cycloaliphatic optionally substituted with one or more Rc. In some embodiments, G3is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, optionally substituted with one or more Rc.

[0110] In some embodiments, G3is 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, optionally substituted with one or more Rc. In some embodiments, G3is 4- to 6-membered monocyclic heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, optionally substituted with one or more Rc. In some embodiments, G3is 6- to 9-membered bicyclic heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, optionally substituted with one or more Rc. In some embodiments, G3is pyrrolidine, piperazine, tetrahydropyran, or piperdine, each optionally substituted with one or more Rc. In some embodiments, G3is 4- to 6-membered monocyclic heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, optionally substituted with optionally substituted C1-C6aliphatic. In some embodiments, G3is 4- to 6-membered monocyclic heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, optionally substituted with C1-C6aliphatic, said C1- C6aliphatic being optionally substituted with –(CH2)0–4C(O)NR°2or –(CH2)0–4C(O)R°, wherein R° is C1–6aliphatic or 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, optionally substituted with –(CH2)0–2R●.

[0111] In some embodiments, G3is C1-C6aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, optionally substituted with one or more Rc. In some embodiments, G3is C1-C6aliphatic-C(O)- 4- to 6-membered monocyclic heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, optionally substituted with one or more Rc. In some embodiments, G3is C1-C6aliphatic-C(O)- 6- to 9-membered bicyclic heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, optionally substituted with one or more Rc.

[0112] In some embodiments of formula II-1 or III-1, G3is selected from:, wherein --- represents a point of attachment to moiety B.

[0113] In some embodiments of formula II-1 or III-1, G3is selected from:

[0114] In some embodiments of formula II-2 or III-2, G3is selected from:

[0115] In some embodiments of formula II-2 or III-2, G3is selected from:.

[0116] As described herein, each Rais independently selected from H and optionally substituted C1-C6aliphatic. In some embodiments, Rais H. In some embodiments, Rais optionally substituted C1-C6aliphatic.

[0117] As described herein, each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen. In some embodiments, Rbis –N(Ra)2. In some embodiments, Rbis optionally substituted C1-C6aliphatic. In some embodiments, Rbis halogen.

[0118] As described herein, each Rcis independently selected from optionally substituted C1- C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa. In some embodiments, Rcis optionally substituted C1-C6aliphatic. In some embodiments, Rcis optionally substituted C3-C6cycloaliphatic. In some embodiments, Rcis optionally substituted C6-C12aryl. In some embodiments, Rcis optionally substituted 4- to 12- membered heteroaryl. In some embodiments,Rcis optionally substituted 4- to 6-membered heterocycle. In some embodiments, Rcis -ORa. In some embodiments, Rcis -C(O)-C1-C6aliphatic. In some embodiments, Rcis -C(O)-ORa.

[0119] As described herein, B is a linker moiety. In some embodiments, B is a linker moiety that is a bivalent moiety covalently bonded to G1(in formula II-2 and III-2) or G3(in formula II-1 and III-1), on one end, and to moiety C on the other. In some embodiments, B is optionally substituted C2-30aliphatic group wherein one or more carbons are optionally and independently replaced by -Cy-, -NRZ-, -N(RZ)C(O)-, -C(O)N(RZ)-, -N(RZ)C(O)O-, -OC(O)N(RZ)-, - N(RZ)C(O)N(RZ) -, -C(O)N(RZ)SO2(RZ)-, -SO2(RZ)N(RZ)C(O)-,-OC(O)O-, -O-, -C(O)-, -OC(O)- , -C(O)O-, -SO-, -SO2-; where each -Cy- is independently an optionally substituted 3-12 membered bivalent C3-C6cycloaliphatic, 5- to 12-membered heterocyclyl ring having 1-3 heteroatoms selected from N, O, S, 5- to 6-membered heteroaryl ring having 1-3 heteroatoms selected from N, O, S; and each RZis independently H or an optionally substituted group selected from C1-C20aliphatic, or C3-C12cycloaliphatic.

[0120] In some embodiments, B is an optionally substituted C2-C10aliphatic group, an optionally substituted 2- to 10-membered heteroaliphatic group, -C2-C10aliphatic-Cy-, or -C2-C10aliphatic-C(O)-. In some embodiments, B is an optionally substituted C2-C10aliphatic group, or an optionally substituted 2- to 10-membered heteroaliphatic group. In some embodiments, B is an optionally substituted C2-C10aliphatic group. In some embodiments, B is an optionally substituted 2- to 10-membered heteroaliphatic group.

[0121] In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are replaced by -O-. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are replaced by -C(O)-. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -Cy- wherein -Cy- is an optionally substituted C3-C6cycloalkyl. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -Cy- wherein -Cy- is an optionally substituted 6 membered heteroaryl ring having 1 nitrogen heteroatom.

[0122] In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -O- and / or -C(O)-. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one ormore carbons are independently replaced by -O- and / or -N(RZ)C(O)-, wherein RZis H. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -O- and / or -C(O)N(RZ)-, wherein RZis H. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -N(RZ)C(O)- and / or -C(O)- and RZis H. In some embodiments, B is a linker that is is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -C(O)N(RZ)-and / or -C(O)- and RZis H. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -Cy- and / or -C(O)-, wherein -Cy- is C6cycloalkyl and / or a 6 membered bivalent heterocyclyl ring having 2 nitrogen heteroatoms.

[0123] In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -O-, -N(RZ)C(O)-, and / or -Cy- , wherein -Cy- is a substituted 3-12 membered bivalent heteroaryl ring having 1-4 nitrogen heteroatoms and RZis H. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -O-, -C(O)N(RZ)-, and / or -Cy-, wherein -Cy- is a substituted 3-12 membered bivalent heteroaryl ring having 1-4 nitrogen heteroatoms and RZis H. . In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by - N(RZ)C(O)-, -O-, and / or -C(O)- and RZis H. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -C(O)N(RZ)-, -O-, and / or -C(O)- and RZis H.

[0124] In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -O-, -N(RZ)C(O)-, -C(O)-, and / or -Cy-, wherein -Cy- is C6cycloalkyl and RZis H. In some embodiments, B is a linker that is an optionally substituted C2-30aliphatic group wherein one or more carbons are independently replaced by -O-, -C(O)N(RZ)-, -C(O)-, and / or -Cy-, wherein -Cy- is C6cycloalkyl and RZis H.

[0125] In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein one carbon is replaced by -C(O)-. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein one or more carbons are replaced by -C(O)-. In some embodiments, B is a linker that is an optionally substituted C1-6aliphatic group wherein twocarbons are replaced by -C(O)-. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein one or more carbons are independently replaced by -N(RZ)C(O)- wherein RZis H. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein one or more carbons are independently replaced by -C(O)N(RZ)-wherein RZis H. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein one carbon is replaced by -Cy- wherein -Cy- is a 6 membered bivalent heterocyclyl ring having 1 nitrogen heteroatom. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein one carbon is replaced by -Cy- wherein -Cy- is a 6 membered bivalent heterocyclyl ring having 2 nitrogen heteroatoms.

[0126] In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein two carbons are replaced by -O-. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein one or more carbons are independently replaced by - C(O)- and / or -Cy- wherein -Cy- is an optionally substituted C3-C6cycloalkyl. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein two carbons are independently replaced by -C(O)- and -Cy- wherein -Cy- is an optionally substituted 3-12 membered bivalent heteroaryl ring having 1-4 nitrogen heteroatoms. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein two carbons are independently replaced by -C(O)- and -Cy- wherein -Cy- is an optionally substituted 6 membered bivalent heteroaryl ring having 1 nitrogen heteroatom.. In some embodiments, B is a linker that is an optionally substituted C1-10aliphatic group wherein one or more carbons are independently replaced by -C(O)- and / or -Cy- wherein -Cy- is an optionally substituted C7cycloalkyl.

[0127] In some embodiments, moiety A is derived from a compound in Table A: Table A

[0128] In some embodiments, B is a linker moiety selected from Table B: Table B , , , , ,, , , , ,,wherein t is an integer between 1and 20.

[0129] In some embodiments, B is a linker moiety selected from:, wherein t is an integer between 1 and 20.

[0130] As defined generally above, C is a target binding moiety. In some embodiments, C is a target binding moiety that binds to a kinase, a kinase fusion, a protein of interest and / or a fusion protein. In some embodiments, C is a target binding moiety that binds to ALK, BRD4, EML4, Myddosome (IRAK4), Myddosome (MALT1), FGFR, RET, HTT, Tau, NLRP3 Inflammasome, EGFR / RTK, Androgen Receptor, ACC2, KMO, IAPP, TSPO, STING, cGAS, or USP30. In some embodiments, C is a target binding moiety that binds to ALK, Myddosome (IRAK4), MALT1, BRD4, FGFR, or RET.

[0131] In some embodiments, C is a target binding moiety that binds to ALK. In some embodiments, C is a target binding moiety that binds to ALK and is derived from a compound described in WO2017053657, WO2010143664, J Med Chem.2015, 10;58(23):9296-9308, J. Med.Chem. 2019, 62, 10927-10954, ACS Omega, Vol 7, 2022, 32442-32456, KR1691536, W02012106540, W02006021881, US7964592, Clinical Cancer Research (2015), 21(11), 2436- 2439, Journal of Oncology Pharmacy Practice (2017), 23(8), 602-614, WO2009143389, Drugs (2021), 81(2), 267-275, New England Journal of Medicine (2020), 383(21), 2018-2029, WO2013132376, W02017004342, Cancer Discovery (2018), 8(10), 1227-1236, W02020069106, Mol Cancer Then 2021 Sep;20(9):1499-1507, EMBO Molecular Medicine 14: e!4296 (2022), Acta Pharmaceutica Sinica Bl 1(2), 2021, 355-372, J Med Chem. 2016, 14;59(7):3392-408, or Cancer Lett. 2016, 28;372(2): 179-86.

[0132] In some embodiments, C is a target binding moiety that binds to ALK and is derived from a compound selected from:

[0133] In some embodiments, C is a target binding moiety that binds to ALK and is selected from:

[0134] In some embodiments, C is a target binding moiety that binds to BRD4. In some embodiments, C is a binding moiety that binds to BRD4 and is derived from a compound described in Nature.2010 Dec 23;468(7327):1067-73. In some embodiments, C is a target binding moiety that binds to BRD4 and is derived from a compound:

[0135] In some embodiments, C is a target binding moiety that binds to BRD4 and is:

[0136] In some embodiments, C is a target binding moiety that binds to Myddosome (IRAK4). In some embodiments, C is a target binding moiety that binds to Myddosome (IRAK4) and is derived from a compound described in WO2011043371, WO2019133531, WO2015068856, US20180111917, WO2019160915, WO2018098367, Bioorg Med Chem Lett 2006;16(11):2842– 5, ACS MedChem Lett., 2019, 10, p1081, WO2015103453, Cell ChemBio, 2020, 27, p11-10, WO2018060174, ACS MedChem Lett., 2021, 12, p82, WO2019099926, Bioorganic & Medicinal Chemistry Letters (2014), 24(9), 2066-2072, Bioorganic & Medicinal Chemistry Letters (2008), 18(11), 3211-3214, WO2021018118, or WO2022147465.

[0137] In some embodiments, C is a target binding moiety that binds to Myddosome (IRAK4) and is derived from:

[0138] In some embodiments, C is a target binding moiety that binds to Myddosome (IRAK4) and is selected from:

[0139] In some embodiments, C is a target binding moiety that binds to Myddosome (MALT1). In some embodiments, C is a target binding moiety that binds to Myddosome (MALT1) and is derived from a compound described in J Med Chem., 2020 Dec 10;63(23):14594-14608, J Med Chem. 2020 Dec 10;63(23):14576-14593, Cancer Cell. 2012 Dec 11;22(6):812-24, WO2020111087, or WO2018020474. In some embodiments, C is a target binding moiety that binds to Myddosome (MALT1) and is derived from:

[0140] In some embodiments, C is a target binding moiety that binds to Myddosome (MALT1) that is selected from:

[0141] In some embodiments, C is a target binding moiety that binds to FGFR. In some embodiments, C is a target binding moiety that binds to FGFR and is derived from a compound described in WO2008075068, WO2006000420, Cancer Discovery, 2018, 8(3):354-369, Mol Cancer Ther. 2017, 16(6) 1010-1020, Oncotarget, 20167: 24252-24268, WO2013108809, or US20160136168. In some embodiments, C is a target binding moiety that binds to FGFR, and is derived from:.

[0142] In some embodiments, C is a target binding moiety that binds to FGFR and is selected from: ,

[0143] In some embodiments, C is a target binding moiety that binds to RET. In some embodiments, C is a target binding moiety that binds to RET and is derived from a compound described in ACS Med. Chem. Lett.2021, 12, 12, Mol. Cancer Ther.2021, 20 (12_Supplement): P233 (PDB: 7DUA), WO2018071447, Int. J. Mol. Sci.2021, 22(4), 1887, or WO2017079140. In some embodiments, C is a target binding moiety that binds to RET and is derived from:.

[0144] In some embodiments, C is a target binding moiety that binds to RET and is selected

[0145] In some embodiments, C is a target binding moiety that binds to HTT. In some embodiments, C is a target binding moiety that binds to HTT and is derived from a compound described in ACS Med Chem Lett 2020, 63, 8608-8633, WO2020176424, WO2021127265, or ACIE 2017, 56, 11530-11533. In some embodiments, C is a target binding moiety that binds to HTT and is derived from:

[0146] In some embodiments, C is a target binding moiety that binds to HTT and is selected from:

[0147] In some embodiments, C is a target binding moiety that binds to Tau. In some embodiments, C is a target binding moiety that binds to Tau and is derived from a compound described in eLife 2019; 8:e45457, WO2021011913, JMC 2016, 59, 4778-4789, E J Nucl Med Mol Imag 2019, 46, 2178-2189, or JMC 2019, 62, 2974-2987. In some embodiments, C is a target binding moiety that binds to Tau and is derived from:.

[0148] In some embodiments, C is a target binding moiety that binds to Tau and is selected from:

[0149] In some embodiments, C is a target binding moiety that binds to NLRP3Inflammasome. In some embodiments, C is a target binding moiety that binds to NLRP3 Inflammasome and is derived from a compound described in WO2019092170. In some embodiments, C is a target binding moiety that binds to NLRP3 Inflammasome and is selected from:.

[0150] In some embodiments, C is a target binding moiety that binds to EGFR / RTK. In some embodiments, C is a target binding moiety that binds to EGFR / RTK and is derived from a compound described in Cell Chem Bio 2018, 25, 67-77. In some embodiments, C is a target binding moiety that binds to EGFR / RTK and is selected from:

[0151] In some embodiments, C is a target binding moiety that binds to the androgen receptor. In some embodiments, C is a target binding moiety that binds to the androgen receptor and is derived from a compound described in ACS Med Chem Lett 2020, 11, 1539-1547 or WO2018071606. In some embodiments, C is a target binding moiety that binds to the androgen receptor and is selected from:.

[0152] In some embodiments, C is a target binding moiety that binds to ACC2. In some embodiments, C is a target binding moiety that binds to ACC2 and is derived from a compound described in WO201307169 or BOMCL 2011, 21, 6314-6318. In some embodiments, C is a target binding moiety that binds to ACC2 and is derived from:.

[0153] In some embodiments, C is a target binding moiety that binds to ACC2 and is selected from:.

[0154] In some embodiments, C is a target binding moiety that binds to KMO. In some embodiments, C is a target binding moiety that binds to KMO and is selected from a compound described in JMC 2017, 60, 3383-3404. In some embodiments, C is a target binding moiety that binds to KMO and is derived from a compound:

[0156] In some embodiments, C is a target binding moiety that binds to IAPP. In some embodiments, C is a target binding moiety that is derived from a compound described in Diabetologica 2018, 61, 2215-2224. In some embodiments, C is a target binding moiety that binds to IAPP and is derived from a compound:

[0157] In some embodiments, C is a target binding moiety that binds to IAPP and is:

[0158] In some embodiments, C is a target binding moiety that binds to TSPO. In some embodiments, C is a target binding moiety that binds to TSPO and is derived from a compound described in JMC 2017, 60, 7897-7909, Life Sci 1983, 32, 1849-1856, J. Pharmacol. Exp. Ther. 1992, 262, 971−978, or JMC 2015, 58, 7449-7464. In some embodiments, C is a target binding moiety derived from:.

[0159] In some embodiments, C is a target binding moiety that binds to TSPO and is selected from:.

[0160] In some embodiments, C is a target binding moiety that binds to STING. In some embodiments, C is a target binding moiety that binds to STING and is derived from a compound described in ACS Med Chem Lett 2019, 10, 92-97, WO2020132582, or Science 2020, 369, 6506, eaba6098. In some embodiments, C is a target binding moiety that binds to STING and is derived from:

[0161] In some embodiments, C is a target binding moiety that binds to STING and is selected from:

[0162] In some embodiments, C is a target binding moiety that binds to cGAS. In some embodiments, C is a target binding moiety that binds to cGAS and is derived from a compound described in Nat Commun 10, 2261, 2019 or JOC 2020, 85, 1579. In some embodiments, C is a target binding moiety that binds to cGAS and is derived from:

[0163] In some embodiments, C is a target binding moiety that binds to cGAS and is selected from: ,

[0164] In some embodiments, C is a target binding moiety that binds to USP30. In some embodiments, C is a target binding moiety that binds to USP30 and is derived from a compound described in WO2020212350. In some embodiments, C is a target binding moiety that binds to USP30 and is derived from:.

[0165] In some embodiments, C is a target binding moiety that binds to USP30 and is selected from:.

[0166] In some embodiments, C is a target binding moiety that binds to p62. In some embodiments, C is a target binding moiety that binds to p62 and is derived from a compound described in WO2020022783 or Nat Commun.2017 Jul 24;8(1):102. In some embodiments, C is a target binding moiety that binds to p62 and is selected from:.

[0167] In some embodiments, C is a target binding moiety that binds to p62 and is selected from:.

[0168] In some embodiments, C is C1-C20aliphatic substituted with halogen. In some embodiments, C is C1-C20aliphatic-Cl.

[0169] In some embodiments, a compound described herein is a compound of formula IVa:or a pharmaceutically acceptable salt thereof, wherein B, C, Ra, Rb, and G3are as described in classes and subclasses herein.

[0170] In some embodiments, a compound described herein is a compound of formula IVb:or a pharmaceutically acceptable salt thereof, wherein B, C, Ra, and G3are as described in classes and subclasses herein.

[0171] In some embodiments, a compound described herein is a compound of formula IVc-1or a pharmaceutically acceptable salt thereof, wherein B, C, Ra, and G3are as described in classes and subclasses herein.

[0172] In some embodiments, a compound described herein is a compound of formula IVc-2or a pharmaceutically acceptable salt thereof, wherein B, C, Ra, and G3are as described in classes and subclasses herein.

[0173] In some embodiments, a compound described herein is a compound of formula IVd-1or a pharmaceutically acceptable salt thereof, wherein B, C, Ra, and G3are as described in classes and subclasses herein.

[0174] In some embodiments, a compound described herein is a compound of formula IVd-2or a pharmaceutically acceptable salt thereof, wherein B, C, Ra, and G3are as described in classes and subclasses herein.

[0175] In some embodiments, a compound described herein is a compound of formula IVe-1or a pharmaceutically acceptable salt thereof, wherein B, C, Ra, Rb, and G3are as described in classes and subclasses herein.

[0176] In some embodiments, a compound described herein is a compound of formula IVe-2or a pharmaceutically acceptable salt thereof, wherein B, C, Ra, and G3are as described in classes and subclasses herein.

[0177] In some embodiments, a compound described herein is a compound of formula IVfor a pharmaceutically acceptable salt thereof, wherein B, C, Ra, and G3are as described in classes and subclasses herein.

[0178] In some embodiments, a compound described herein is a compound of formula IVgor a pharmaceutically acceptable salt thereof, wherein B, C, Ra, and G3are as described in classes and subclasses herein.

[0179] In some embodiments, a compound described herein is a compound of formula IVhor a pharmaceutically acceptable salt thereof, wherein B, C, Ra, and G3are as described in classes and subclasses herein, and each of W1, W2, W3, and W4is independently selected from N, CH, and CRb, and wherein Rbis described in classes and subclasses herein.

[0180] In some embodiments, a compound described herein is a compound of formula IVior a pharmaceutically acceptable salt thereof, wherein B, C, Ra, Rb, and G3are as described in classes and subclasses herein.

[0181] In some embodiments, a compound described herein is a compound of formula IVjor a pharmaceutically acceptable salt thereof, wherein RaG3are as described in classes and subclasses herein, and each of W5, W6, W7, and W8is independently selected from N and CRd,wherein each Rdis indepdnently selected from H, –N(Ra)2, optionally substituted C1-C6aliphatic, halogen, or moiety B-C wherein one of W1, W2, W3or W4is CRd, and Rdis moiety B-C,.

[0182] In some embodiments, a compound of formula I-IVj is selected from Table 1, or a pharmaceutically acceptable salt thereof: Table 1

[0183] In some embodiments, a compound of formula I is selected from:

[0184] In some embodiments, a compound that binds p62 is a compound is of formula X:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen;each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa.

[0185] In some embodiments, a compound that binds p62 is a compound is of formula XI:or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, - ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa.

[0186] In some embodiments of formulae X or XI, any of variables Ring A, G1, G2, G3, Ra, Rb, and Rcare as defined classes and subclasses with respect to in formula I herein.

[0187] In some embodiments, a compound of formula X or XI is selected from Table 2, or a pharmaceutically acceptable salt thereof: Table 2

[0188] In some embodiments, provided compounds are provided and / or utilized in a salt form (e.g., a pharmaceutically acceptable salt form). Reference to a compound provided herein is understood to include reference to salts thereof, unless otherwise indicated.Uses, Formulation, and Administration Pharmaceutically Acceptable Compositions

[0189] According to another embodiment, the present disclosure provides a composition comprising a compound described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. In certain embodiments, a composition described herein is formulated for administration to a patient in need of such composition. In some embodiments, a composition described herein is formulated for oral administration to a patient.

[0190] Compounds and compositions, according to method of the present disclosure, are administered using any amount and any route of administration effective for treating or lessening the severity of a disorder provided herein. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the particular agent, its mode of administration, and the like. Compounds described herein are preferably formulated in unit dosage form for ease of administration and uniformity of dosage.

[0191] Compositions of the present disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, intraperitoneally, intracisternallyor via an implanted reservoir. In some embodiments, the compositions are administered orally, intraperitoneally or intravenously.

[0192] Sterile injectable forms of the compositions described herein may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

[0193] For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similardispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.

[0194] Injectable formulations can be sterilized, for example, by filtration through a bacterial- retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0195] In order to prolong the effect of a compound of the present disclosure, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide- polyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

[0196] In some embodiments, provided pharmaceutically acceptable compositions are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions described herein are administered without food. In other embodiments, pharmaceutically acceptable compositions described herein are administered with food. Pharmaceutically acceptable compositions described herein may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient iscombined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.

[0197] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and / or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and / or i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0198] Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.

[0199] The active compounds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch.Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0200] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0201] Alternatively, pharmaceutically acceptable compositions described herein may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

[0202] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0203] Pharmaceutically acceptable compositions described herein may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

[0204] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.

[0205] For topical applications, provided pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of compounds described herein include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0206] For ophthalmic use, provided pharmaceutically acceptable compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutically acceptable compositions may be formulated in an ointment such as petrolatum.

[0207] Pharmaceutically acceptable compositions described herein may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well- known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0208] Dosage forms for topical or transdermal administration of a compound disclosed herein include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of this disclosure. Additionally, the present disclosure contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel. Diseases, Disorders, and Conditions

[0209] The present disclosure encompasses an insight that compounds and compositions provided herein can be used to direct autophagy to a target. The present disclosure also encompasses an insight that directing autophagy to particular targets can be useful for treating particular diseases, disorders, and conditions. In some embodiments, a disease, disorder, or condition is disease, disorder, or condition is selected from NASH, NAFLD, cancer (e.g., cervical cancer, colon cancer, breast cancer, lung cancer, stomach cancer, gastrointestinal cancer, pancreatic cancer, prostate cancer, leukemia, melanoma, lymphoma), Burkitt lymphoma, active B- cell-like diffuse large B-cell lymphomas, diffuse large B-cell lymphomas, primary central nervous system lymphomas, IgM-secreting lymphoplasmacytic lymphoma, Waldenstrom macroglobulinemia, gout, atherosclerosis, Alzheimer’s disease, diabetes (e.g., Type II diabetes), experimental autoimmune encephalitis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, cryopyrin-associated periodic syndromes, Parkinson’s disease, Dementia with Lewy bodies, multiple systems atrophy, neuroaxonal dystrophies, primary age-related tauopathy (PART) dementia, chronic traumatic encephalopathy, Progressive supranuclear palsy (PSP), Corticobasal degeneration (CBD), Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), Lytico-bodig disease (Parkinson-dementia complex of Guam), Ganglioglioma and gangliocytoma, Meningioangiomatosis, Postencephalitic parkinsonism, Subacute sclerosing panencephalitis (SSPE), Lead encephalopathy, Tuberous sclerosis, Pantothenate kinase-associated neurodegeneration, Lipofuscinosis, Spinal and bulbar muscular atrophy (SBMA) / Kennedy’s disease, rheumatoid arthritis, psoriasis, Systemic lupus erythematosus, Aicardia-Goutieres syndrome, ataxia, Familial chilblain lupus, Huntington’s disease, Spinocerebellar ataxia, Familial amylotrophic lateral sclerosis, Frontotemporal dementia (FTLD-TDP), and Amyotrophic lateral sclerosis.

[0210] In some embodiments, a particular target binding moiety (e.g, moiety C’ in formula I, above) can be selected to modulate (and thereby induce degradation of) a target of choice to to treat a disease, disorder, or condition of choice. A person of skill in the art can identify particularbinding moieties based on the desired disease, disorder, or condition to be treated, using known binding moieties.

[0211] In some embodiments, a target is a lipid droplet, and a disease, disorder or condition to be treated is selected from NASH and NAFLD.

[0212] In some embodiments, a target is COP9, and a disease, disorder or condition is cancer.

[0213] In some embodiments, a target is MYC, and a disease, disorder or condition is Burkitt lymphoma, cervical cancer, colon cancer, breast cancer, lung cancer, or stomach cancer.

[0214] In some embodiments, a target is myddosome, and a disease, disorder or condition is active B-cell-like diffuse large B-cell lymphomas (ABC DLBCL), diffuse large B-cell lymphomas, primary central nervous system lymphomas, IgM-secreting lymphoplasmacytic lymphoma, or Waldenstrom macroglobulinemia.

[0215] In some embodiments, a target is inflammasome, and a disease, disorder or condition is gout, atherosclerosis, Alzheimer’s disease, Type-II diabetes, experimental autoimmune encephalitis, multiple sclerosis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, cryopyrin-associated periodic syndromes (CAPS), colon cancer, breast cancer, melanoma, hepatitis C virus-associated hepatocellular carcinoma, and gastrointestinal cancers.

[0216] In some embodiments, a target is KRAS, and a disease, disorder or condition is colorectal cancer, lung cancer, leukemia, pancreatic cancer,

[0217] In some embodiments, a target is α-synuclein, and a disease, disorder or condition is Parkinson’s disease, dementia with Lewy bodies, multiple systems atrophy, or neuroaxonal dystrophies.

[0218] In some embodiments, a target is tau, and a disease, disorder or condition is Alzheimer’s disease, Primary age-related tauopathy (PART) dementia, chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), Lytico-bodig disease (Parkinson-dementia complex of Guam), ganglioglioma, gangliocytoma, meningioangiomatosis, postencephalitic parkinsonism, subacute sclerosing panencephalitis (SSPE), lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, or lipofuscinosis.

[0219] In some embodiments, a target is estrogen receptor-α, and a disease, disorder or condition is breast cancer.

[0220] In some embodiments, a target is an androgen receptor, and a disease, disorder or condition is prostate cancer or spinal and bulbar muscular atrophy (SBMA) / Kennedy’s disease.

[0221] In some embodiments, a target is KSR1, and a disease, disorder or condition is cancer.

[0222] In some embodiments, a target is islet amyloid polypeptide (IAPP), and a disease, disorder or condition is type-II diabetes.

[0223] In some embodiments, a target is IRAK4, and a disease, disorder or condition is rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), or psoriasis.

[0224] In some embodiments, a target is PINK1, and a disease, disorder or condition is Parkinson’s disease, Huntingtin’s disease, or spinocerebellar ataxia.

[0225] In some embodiments, a target is TDP43, and a disease, disorder or condition is familial amylotrophic lateral sclerosis or frontotemporal dementia (FTLD-TDP). Exemplary Embodiments

[0226] The present disclosure provides the following non-limiting numbered embodiments. Embodiment 1. A compound of formula I: A-B-C I or a pharmaceutically acceptable salt thereof, wherein A is a moiety that binds or associated with p62; B is a linker moiety; and C is a target binding moiety. Embodiment 2. The compound of Embodiment 1, wherein A is a polypeptide or peptidomimetic moiety that binds or associates with p62. Embodiment 3. The compound of Embodiments 1 or 2, wherein A is a dipeptide or peptidomimetic moiety that binds or associates with p62. Embodiment 4. The compound of Embodiment 1, wherein the compound is of formula II-1 or II-2:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, - C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety. Embodiment 5. The compound of Embodiment 1, wherein the compound is of formula III- 1 or III-2:or a pharmaceutically acceptable salt thereof, wherein:Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, - C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6- membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety. Embodiment 6. The compound of Embodiments 4 or 5, wherein G1is optionally substituted 5- to 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S. Embodiment 7. The compound of Embodiments 4 or 5, wherein G1is 5- to 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S, optionally substituted with one or more of –N(Ra)2, or halogen.Embodiment 8. The compound of Embodiments 4 or 5, wherein G1is 7 to 12-membered bicyclic heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, optionally substituted with one or more –N(Ra)2, or halogen. Embodiment 9. The compound of any one of Embodiments 4-8, wherein the compound is of formula II-1 or III-1. Embodiment 10. The compound of Embodiment 9, wherein G1is selected from:Embodiment 11. The compound of any one of Embodiments 4-8, wherein the compound is of formula II-2 or III-2. Embodiment 12. The compound of Embodiment 11, wherein G1is selected from:wherein represents a point of attachment to moiety B, and represents a point of attachment to moiety G2.Embodiment 13. The compound of any one of Embodiments 4-12, wherein G2is C1-C6aliphatic or C1-C6aliphatic-4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, wherein G2is optionally substituted with one or more Rb. Embodiment 14. The compound of Embodiment 13, wherein G2is C1-C6aliphatic optionally substituted with one or more Rb. Embodiment 15. The compound of Embodiment 14, wherein G2is C1-C6aliphatic optionally substituted with one or more –N(Ra)2. Embodiment 16. The compound of Embodiment 15, wherein G2is C1-C6aliphatic, substituted with –NH2. Embodiment 17. The compound of Embodiment 16, wherein G2is C1-C6aliphatic, substituted with –NH2, wherein the carbon atom bonded to –NH2is in an S enantiomeric configuration. Embodiment 18. The compound of Embodiment 13, wherein G2is C1-C6aliphatic-4- to 6- membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, wherein G2is optionally substituted with one or more Rb. Embodiment 19. The compound of Embodiment 18, wherein G2is –CH2-4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S. Embodiment 20. The compound of any one of Embodiments 4-12, wherein G2is selected from:whereinrepresents a point of attachment to –C(O)-N(Ra)- in formulae II-1 or II-2, or to A in formulae III-1 or III-2, and represents a point of attachment to G1. Embodiment 21. The compound of any one of Embodiments 4-20, wherein G3is C1-C7aliphatic-C(O)N(Ra)- or 4- to 7-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, wherein G3is optionally substituted with one or more Rc. Embodiment 22. The compound of Embodiment 21, wherein G3is C1-C7aliphatic- C(O)N(Ra) optionally substituted with halogen or an optionally substituted C1-C6aliphatic. Embodiment 23. The compound of Embodiment 22, wherein G3is C1-C7alkyl-C(O)N(Ra) optionally substituted with halogen or an optionally substituted C1-C6aliphatic. Embodiment 24. The compound of Embodiments 9, 10, or 13-20, wherein G3is selected from:wherein - represents a point of attachment to moiety B.-- Embodiment 25. The compound of any one of Embodiments 11-20, wherein G3is selected from:Embodiment 26. The compound of Embodiment 24, wherein G3is selected from:wherein --- represents a point ofattachment to moiety B. Embodiment 27. The compound of Embodiment 25, wherein G3is selected from:. Embodiment 28. The compound of any one of Embodiments 5-27, wherein Ring A is :Embodiment 29. The compound of Embodiment 4, wherein the compound is of formula IVa:or a pharmaceutically acceptable salt thereof. Embodiment 30. The compound of Embodiment 4, wherein the compound is of formula IVbor a pharmaceutically acceptable salt thereof. Embodiment 31. The compound of Embodiment 4, wherein the compound is of formula IVc- 1:or a pharmaceutically acceptable salt thereof. Embodiment 32. The compound of Embodiment 4, wherein the compound is of formula IVd- 1:or a pharmaceutically acceptable salt thereof. Embodiment 33. The compound of Embodiment 4, wherein the compound is of formula IVe- 1:or a pharmaceutically acceptable salt thereof. Embodiment 34. The compound of Embodiment 4, wherein the compound is of formula IVf:or a pharmaceutically acceptable salt thereof. Embodiment 35. The compound of Embodiment 4, wherein the compound is of formula IVg:or a pharmaceutically acceptable salt thereof. Embodiment 36. The compound of Embodiment 4, wherein the compound is of formula IVhor a pharmaceutically acceptable salt thereof, wherein ach of W1, W2, W3, and W4is independently selected from N, CH, and CRb. Embodiment 37. The compound of Embodiment 4, wherein the compound is of formula IVior a pharmaceutically acceptable salt thereof. Embodiment 38. The compound of Embodiment 4, wherein the compound is of formula IVjor a pharmaceutically acceptable salt thereof, wherein each of W5, W6, W7, and W8is independently selected from N and CRd,wherein each Rdis indepdnently selected from H,–N(Ra)2, optionally substituted C1-C6aliphatic, halogen, or moiety B-C wherein one of W1, W2, W3or W4is CRd, and Rdis moiety B-C. Embodiment 39. The compound of any one of Embodiments 1-38, wherein the linker is an optionally substituted C2-30aliphatic group wherein one or more carbons are optionally and independently replaced by -Cy-, -NRZ-, -N(RZ)C(O)-, -C(O)N(RZ)-, -N(RZ)C(O)O-, - OC(O)N(RZ)-, -N(RZ)C(O)N(RZ) -, -OC(O)O-, -O-, -C(O)-, -OC(O)-, - C(O)O-, -SO-, -SO2-, wherein each -Cy- is independently an optionally substituted 3-12 membered bivalent heterocyclyl ring having 1-3 heteroatoms selected from N, O, and S, an optionally substituted 3-8 membered bivalent heteroaryl ring having 1-4 heteroatoms selected from N, O, and S, an optionally substituted C3-C6cycloalkyl, or an optionally substituted C6-C12aryl, and each RZis independently H or an optionally substituted group selected from C1-C20aliphatic, or C3-C12cycloaliphatic. Embodiment 40. The compound of any one of Embodiments 1-39, wherein B is a linker moiety selected from Table B. Embodiment 41. The compound of any one of Embodiments 1-40, wherein C is a is a target binding moiety that binds to ALK, BRD4, p62, Myddosome (IRAK4), Myddosome (MALT1), FGFR, RET, HTT, EML4, Tau, NLRP3 Inflammasome, EGFR / RTK, Androgen Receptor, ACC2, KMO, IAPP, TSPO, STING, cGAS, or USP30. Embodiment 42. The compound of Embodiment 1, wherein the compound is selected from Table 1. Embodiment 43. A pharmaceutical composition comprising a compound of any one of Embodiments 1-42, and a pharmaceutically acceptable carrier, filler, or diluent. Embodiment 44. A method of treating a disease, disorder, or condition in a subject comprising administering a compound of any one of Embodiments 1-42, or the pharmaceutical composition of Embodiment 43. Embodiment 45. The method of Embodiment 44, wherein the disease, disorder, or condition is selected from NASH, NAFLD, cancer (e.g., cervical cancer, colon cancer, breast cancer, lung cancer, stomach cancer, gastrointestinal cancer, pancreatic cancer, prostate cancer, leukemia, melanoma, lymphoma), Burkitt lymphoma, active B-cell-like diffuse large B- cell lymphomas, diffuse large B-cell lymphomas, primary central nervous system lymphomas, IgM-secreting lymphoplasmacytic lymphoma, Waldenstrommacroglobulinemia, gout, atherosclerosis, Alzheimer’s disease, diabetes (e.g., Type II diabetes), experimental autoimmune encephalitis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, cryopyrin-associated periodic syndromes, Parkinson’s disease, Dementia with Lewy bodies, multiple systems atrophy, neuroaxonal dystrophies, primary age-related tauopathy (PART) dementia, chronic traumatic encephalopathy, Progressive supranuclear palsy (PSP), Corticobasal degeneration (CBD), Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), Lytico-bodig disease (Parkinson- dementia complex of Guam), Ganglioglioma and gangliocytoma, Meningioangiomatosis, Postencephalitic parkinsonism, Subacute sclerosing panencephalitis (SSPE), Lead encephalopathy, Tuberous sclerosis, Pantothenate kinase-associated neurodegeneration, Lipofuscinosis, Spinal and bulbar muscular atrophy (SBMA) / Kennedy’s disease, rheumatoid arthritis, psoriasis, Systemic lupus erythematosus, Aicardia-Goutieres syndrome, ataxia, Familial chilblain lupus, Huntington’s disease, Spinocerebellar ataxia, Familial amylotrophic lateral sclerosis, Frontotemporal dementia (FTLD-TDP), and Amyotrophic lateral sclerosis. Embodiment 46. A method of inducing degradation of a target in a biological sample, comprising contacting the biological sample with a compound of any one of Embodiments 1-42, or a pharmaceutical composition of Embodiment 43. Embodiment 47. A compound of any one of Embodiments 1-42, or a pharmaceutical composition of Embodiment 43, for use in medicine. Embodiment 48. Use of a compound of any one of Embodiments 1-42, or a pharmaceutical composition of Embodiment 43, in the treatment of a disease, disorder, or condition. Embodiment 49. The use of Embodiment 48, wherein the disease, disorder, or condition is NASH, NAFLD, cancer (e.g., cervical cancer, colon cancer, breast cancer, lung cancer, stomach cancer, gastrointestinal cancer, pancreatic cancer, prostate cancer, leukemia, melanoma, lymphoma), Burkitt lymphoma, active B-cell-like diffuse large B-cell lymphomas, diffuse large B-cell lymphomas, primary central nervous system lymphomas, IgM-secreting lymphoplasmacytic lymphoma, Waldenstrom macroglobulinemia, gout, atherosclerosis, Alzheimer’s disease, diabetes (e.g., Type II diabetes), experimental autoimmune encephalitis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, cryopyrin-associated periodic syndromes, Parkinson’s disease, Dementia with Lewybodies, multiple systems atrophy, neuroaxonal dystrophies, primary age-related tauopathy (PART) dementia, chronic traumatic encephalopathy, Progressive supranuclear palsy (PSP), Corticobasal degeneration (CBD), Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), Lytico-bodig disease (Parkinson- dementia complex of Guam), Ganglioglioma and gangliocytoma, Meningioangiomatosis, Postencephalitic parkinsonism, Subacute sclerosing panencephalitis (SSPE), Lead encephalopathy, Tuberous sclerosis, Pantothenate kinase-associated neurodegeneration, Lipofuscinosis, Spinal and bulbar muscular atrophy (SBMA) / Kennedy’s disease, rheumatoid arthritis, psoriasis, Systemic lupus erythematosus, Aicardia-Goutieres syndrome, ataxia, Familial chilblain lupus, Huntington’s disease, Spinocerebellar ataxia, Familial amylotrophic lateral sclerosis, Frontotemporal dementia (FTLD-TDP), and Amyotrophic lateral sclerosis Embodiment 50. A compound of formula X:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, - C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa. Embodiment 51. A compound of formula XI:or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, - C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc;each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa. Embodiment 52. The compound of Embodiments 50 or 51, wherein the compound is selected from Table 2. EXAMPLES

[0227] As described in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. It will be appreciated that, although the general methods depict the synthesis of certain compounds of the present disclosure, the following general methods and other methods known to one of ordinary skill in the art can be applied to all compounds and subclasses and species of each of these compounds, as described herein. Table of AbbreviationsAnalytical Instrumentation and Purification

[0228] NMR Instrument Details: Varian 400MHz, Probe-1: Auto XID Probe 2: ATB.

[0229] LCMS Instrument Details: Shimadzu LCMS-2010EV system coupled to SPD-M20A PDA and ELS detectors. Softa model 400. LCMS Method 1 - Acidic conditions Column: X-Select C18 CSH (3.0*50) mm 2.5µ; Make: Waters Mobile Phase A: 0.05% formic acid in water: Acetonitrile ( 95:5); pH= 3.5 Mobile Phase B: 0.05% formic acid in Acetonitrile Column oven temperature: 50 C Flow rate: 1.2 ml / minute PDA: 210nm Maxplot Gradient program :MS Parameters Mode: Dual (+ / -) Detector voltage: 1.5KV Scan rang: 80-2000amu Scan speed: 2000LCMS Method 2 - Basic conditions Column: X-Select C18 CSH (3.0*50) mm 2.5µm ; Make: Waters Mobile Phase A : 5mM Ammonium Bicarb; pH= 8.8 Mobile Phase B: Acetonitrile Column oven temperature: 50 C Flow rate: 1.2 ml / minute PDA: 210nm Maxplot Gradient program :MS Parameters Mode: Dual (+ / -) Detector voltage: 1.5KV Scan rang: 80-2000amu Scan speed: 2000 HPLC Method 1 – Acidic Conditions Column : X-Select CSH C18 (4.6*150) mm; 5µ; Make: Waters Mobile Phase: A - 0.1% Formic acid in water : Acetonitrile(95:05) ; pH=3.5 B - Acetonitrile Flow Rate: 1.0. mL / minute PDA : 210nm maxplot Gradient program :HPLC Method 2 – Basic Conditions Column : Xbridge C18 (4.6*150) mm, 5µ; Make: Waters Mobile Phase A - 0.1% NH3 in water; pH=9.5 B – Acetonitrile Flow Rate: 1.2. mL / minute PDA : 210nm maxplot Gradient program :INT-1. Synthesis of (Z)-N2,Nw,Nw'-tris(tert-butoxycarbonyl)-L-arginine

[0230] Step-1. Synthesis of (Z)-N2,Nw,Nw'-tris(tert-butoxycarbonyl)-L-arginine, INT-1

[0231] (2S)-2-amino-5-carbamimidamidopentanoic acid (8.70 g, 49.9 mmol) was added into a solution of t-BuOH (150 mL) and H2O (150 mL) in a 500 mL round-bottom flask. The mixture was cooled to 0 °C in an ice bath and NaOH (6.96 g, 174 mmol) was added. The solution was stirred for 5 min at 0 °C. Then Boc2O (43.30 g, 199.0 mmol) was added to the mixed solution in portions. The reaction mixture was stirred for two days at room temperature. The t-BuOH (150 mL) was evaporated under reduced pressure and the residue was acidified with citric acid to pH=3.The above solution extracted with ethyl acetate (100 mL) for 3 times. The combined organic phase was dried with anhydrous sodium sulfate and evaporated in a vacuum to afford (2S)-2-{[(tert- butoxy)carbonyl]amino}-5-{[(Z)-{[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl] imino})methyl]amino}pentanoic acid, INT-1. Yield: 7.73 g, 33%; Appearance: White solid;1H NMR (300 MHz, DMSO-d6) δ 12.09 (s, 1H), 9.41- 8.85 (m, 2H), 7.06 (J = 8.0 Hz, 1H), 3.94 – 3,73 (m, 3H), 1.61 – 1.53 (m, 2H), 1.49 (s, 9H), 1.41 (s, 9H), 1.39 – 1.26 (m, 11H). HPLC purity: 85.0%; LCMS Calculated for C21H38N4O8: 474.27; Observed: 475.3[M+H]+. INT-2. Synthesis of (2S)-5-(2-aminopyridin-3-yl)-2-{[(tert-butoxy)carbonyl]amino}pentanoic acid

[0232] Step-1. Synthesis of methyl (2S)-5-(2-aminopyridin-3-yl)-2-{[(tert-butoxy) carbonyl]amino}pent-4-ynoate

[0233] To a solution of 3-iodopyridin-2-amine (350 mg, 1.59 mmol) in DMF (2 mL) was added TEA (802 mg, 7.95 mmol), Pd / (PPh3)Cl2(111 mg, 0.16 mmol), CuI (30.2 mg, 0.16 mmol) and methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}pent-4-ynoate (540 mg, 2.38 mmol). The mixture solution was stirred at 80 °C for 1 h under N2. The reaction mixture was poured into water (20 mL), extracted with EA (20 mL x 3). The combined organic layer was washed with brine (10 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by flash silica gel chromatography (EA-PE = 0%-50%) to give methyl (2S)-5-(2-aminopyridin-3- yl)-2-{[(tert-butoxy)carbonyl]amino}pent-4-ynoate (430 mg, 1.34 mmol, 100% purity, 84.8% yield) as yellow oil.

[0234] Step-2. Synthesis of methyl (2S)-5-(2-aminopyridin-3-yl)-2-{[(tert-butoxy) carbonyl] amino} pentanoate

[0235] To a solution of methyl (2S)-5-(2-aminopyridin-3-yl)-2-{[(tert-butoxy) carbonyl]amin o}pent-4-ynoate (430 mg, 1.34 mmol) in MeOH (5 mL) was added Pd / C (426 mg, 4.02 mmol) under N2. The suspension was degassed with H2for 6 times. The mixture was stirred at rt for 16 h under H2. After filtration via Celite pad, the organic layer was concentrated under reduced pressure to dryness to give methyl (2S)-5-(2-aminopyridin-3-yl)-2-{[(tert-butoxy)carbonyl]amino} pentanoate (433 mg, 1.33 mmol, 100% purity, 100% yield) as yellow oil.

[0236] Step-3. Synthesis of (2S)-5-(2-aminopyridin-3-yl)-2-{[(tert- butoxy)carbonyl]amino}pentanoic acid, INT-2

[0237] To a solution of methyl (2S)-5-(2-aminopyridin-3-yl)-2-{[(tert- butoxy)carbonyl]amino} pentanoate (430 mg, 1.32 mmol) in H2O (2 mL) and THF (2 mL) was added LiOH (63.3 mg, 2.64 mmol). The mixture solution was stirred at RT for 1 h. The reaction mixture was concentrated to give (2S)-5-(2-aminopyridin-3-yl)-2-{[(tert-butoxy) carbonyl]amino}pentanoic acid, INT-2 (408 mg, 1.31 mmol, 100% purity, 100% yield) as a yellow solid. INT-3. Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H-imidazol-1-yl)pentanoic acid

[0238] Step-1. Synthesis of Benzyl (S)-2-((tert-butoxycarbonyl)amino)-5-hydroxypentanoate

[0239] To a stirred solution of 2-benzyl 1-tert-butyl 5-oxopyrrolidine-1,2-dicarboxylate (10.0 g, 31.3 mmol, 1.0 eq.) in tetrahydrofuran (120.0 mL) and water (20.0 mL) was added sodium borohydride (2.36 g, 62.6 mmol, 2.0 eq.) at 0 °C. The mixture was stirred for 1 h at 0 °C and allowed to warm to room temperature, then stirred at room temperature for 1 h. LCMS showed the reaction was completed and product was produced. The reaction mixture was concentrated in vacuo and the residue was suspended in EA and washed with water. The aqueous layer was extracted with EA and the combined organic extracts washed with water, brine, dried and concentrated in vacuo to give the crude that was purified by flash chromatography eluting with 0-50% EA in PE to afford benzyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-hydroxypentanoate (3.96 g, 100% purity, 38% yield)) as a colorless oil.

[0240] Step-2. Synthesis of Benzyl (S)-2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H- imidazol-1-yl)pentanoate

[0241] To a solution of benzyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-hydroxypentanoate (3.96 g, 12.2 mmol, 1.0 eq.) in Tetrahydrofuran (40.0 mL ) was added 2-nitro-1H-imidazole (2.75 g, 24.4 mmol, 2.0 eq.) , triphenylphosphine (6.39 g, 24.4 mmol, 2.0 eq.) and diisopropyl azodicarboxylate (4.93 g, 24.4 mmol, 2.0 eq.). The reaction was stirred at room temperature for 16 h. LCMS showed the reaction was completed and the product was produced. The reaction mixture was concentrated to give benzyl (S)-2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H-imidazol-1- yl)pentanoate (13.2 g, crude). The crude was used for next step directly without further purification.

[0242] Step-3. Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H-imidazol-1- yl)pentanoic acid

[0243] To a solution of benzyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H- imidazol-1-yl)pentanoate (13.2 g, 31.5 mmol, crude) in methanol (120.0 mL) was added lithiumol (1.50 g, 63.0 mmol, 2.0 eq.) and water (40.0 mL). The reaction was stirred at room temperature for 16 h. LCMS showed the reaction was completed and evaporated to remove solvent in vacuo. The residue was dissolved in water and EA, the resulting mixture was extracted three times with EA to remove by-products of the previous reaction. The water layer was acidified with 1 N HCl (aq) until pH = 1 and extracted with EA. The combined organic layer was washed with brine, dried over sodium sulfate, filtered, and evaporated under reduced pressure to afford (2S)-2-{[(tert- butoxy)carbonyl]amino}-5-(2-nitro-1H-imidazol-1-yl)pentanoic acid (INT-3). Yield: 3.34 g, 84%; Appearance: White solid;1H NMR (400 MHz, DMSO-d6) δ 12.61 (s, 1H), 7.66 (d, J = 0.8 Hz, 1H), 7.39 (s, 1H), 7.18 (d, J = 0.8 Hz, 1H), 7.11 (d, J = 8.0 Hz, 1H), 4.44 – 4.35 (m, 2H), 3.90 – 3.84 ( m, 1H), 1.95 – 1.75 (m, 2H), 1.72 – 1.64 (m, 1H), 1.56 – 1.44 (m, 1H), 1.35 (s, 9H); HPLC purity: 100%; LCMS Calculated for C13H20N4O6: 328.14; Observed: 219.1 [M+H]+. INT-4 & INT-9. Synthesis of (2S)-5-(2-amino-1H-1,3-benzodiazol-1-yl)-2-{[(tert-butoxy) carbonylamino}pentanoic acid.

[0244] Step-1. Synthesis of 1-benzyl 2,5-dioxopyrrolidin-1-yl (2S)-2-{[(tert- butoxy)carbonyl]amino} pentanedioate

[0245] Into a 500 mL 3-necked round-bottom was placed (4S)-5-(benzyloxy)-4-{[(tert- butoxy)carbonyl]amino}-5-oxopentanoic acid (10.0 g, 29.6 mmol) and NHS (3.73 g, 32.5 mmol) in DCM (150 mL) was added DCC (7.27 g, 35.5 mmol) at 0°C. The resulting mixture was stirred overnight at room temperature under nitrogen atmosphere. The reaction mixture was diluted with water (200 mL) and extracted with DCM (300 mLx3). The combined organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuum to afford the crude 1-benzyl 2,5-dioxopyrrolidin-1-yl (2S)-2-{[(tert-butoxy)carbonyl]amino} pentanedioate (11.5 g, 26.5 mmol, 92% purity, 83% yield) as a white solid, which was used for next step directly.

[0246] Step-2. Synthesis of benzyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5- hydroxypentanoate

[0247] Into a 1-L 3-necked round-bottom was placed 1-benzyl 2,5-dioxopyrrolidin-1-yl (2S)- 2-{[(tert-butoxy)carbonyl]amino}pentanedioate (12.0 g, 27.6 mmol) in THF (120 mL), followed by dropwise addition of a solution of NaBH4(2.09 g, 55.2 mmol) in THF / H2O (5:1, 60 mL) at 0°C under nitrogen atmosphere. The resulting mixture was stirred for 1.0 hour at 0°C under nitrogen atmosphere. The resulting mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mLx3). The combined organic phase was washed with sat brine (200 mL), dried over Na2SO4, filtered, and concentrated in vacuum. The residue was purified by silica gel chromatography (Petroleum ether / ethyl acetate =1:1) to give benzyl (2S)-2-{[(tert- butoxy)carbonyl]amino}-5-hydroxypentanoate (7.40 g, 22.8 mmol, 96% purity, 90% yield) as colorless oil.

[0248] Step-3. Synthesis of benzyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H-1,3- benzodiazol-1-yl)pentanoate, INT-4

[0249] To a stirred solution of 2-nitro-1H-1,3-benzodiazole (1.00 g, 6.18 mmol), benzyl (2S)- 2-{[(tert-butoxy)carbonyl]amino}-5-hydroxypentanoate (2.0 g, 6.18 mmol) and PPh3 (2.58 g, 9.88 mmol) in THF (30 mL) was added DIAD (1.87 g, 9.27 mmol) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 2 hours at room temperature. The resulting mixture was concentrated in vacuum, the residue was purified by prep-HPLC (NH3·H2O buffer) to give benzyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H-1,3-benzodiazol-1- yl)pentanoate, INT-4 (2.10 g, 4.48 mmol, 95.6% purity, 72.5% yield) as a yellow solid. LCMS Calculated for C17H22N4O6: 378.385; Observed: 323.1 [(M-t-Bu+H)+].

[0250] Step-4. Synthesis of (2S)-5-(2-amino-1H-1,3-benzodiazol-1-yl)-2-{[(tert- butoxy)carbonyl] amino}pentanoic acid, INT-9

[0251] Into a 100 mL round-bottom flask, was placed benzyl (2S)-2-{[(tert- butoxy)carbonyl]amino}-5-(2-nitro-1H-1,3-benzodiazol-1-yl)pentanoate (2.00 g, 4.26 mmol) in MeOH (30 mL) was added Pd / C (455 mg, 4.26 mmol, 10%) at room temperature under nitrogen atmosphere and then the reaction mixture was degassed and purged with hydrogen for three times. The resulting reaction mixture was stirred for 2 hours under hydrogen atmosphere. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give (2S)-5-(2-amino-1H-1,3- benzodiazol-1-yl)-2-{[(tert-butoxy)carbonyl]amino}pentanoic acid, INT-9. Yield: 1.0 g, 65%; 1H NMR (300 MHz, DMSO-d6) δ 7.35 (s, 1H), 7.14 (t, J = 7.7 Hz, 2H), 6.91 – 6.74 (m, 2H), 6.63 (s, 2H), 4.79 – 4.66 (m, 1H), 3.96 – 3.90 (m, 2H), 1.67 – 1.55 (m, 2H), 1.38 (s, 9H), 1.26 – 1.08 (m, 2H). HPLC purity: 85.2%; LCMS Calculated for C17H24N4O4: 348.18; Observed:349.1[M+H]+. INT-5 & INT-6. Synthesis of 9-ethyl-6,6-dimethyl-11-oxo-8-(piperazin-1-yl)-6,11-dihydro-5H- benzo[b]carbazole-3-carbonitrile (INT-6).

[0252] Step-1. Synthesis of tert-butyl 4-(3-cyano-9-ethyl-6,6-dimethyl-11-oxo-6,11-dihydro- 5H-benzo[b]carbazol-8-yl)piperazine-1-carboxylate (INT-5)

[0253] To a solution of 9-ethyl-8-iodo-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazole- 3-carbonitrile (10.0 g, 22.7 mmol) and tert-butyl piperazine-1-carboxylate (4.63 g, 24.9 mmol) in THF (200 mL) were added Pd2(dba)3(2.07 g, 2.27 mmol), S-Phos (930 mg, 2.27 mmol) and NaHMDS (1M, 90 mL, 90.8 mmol) at room temperature. The resulting mixture was stirred for 3 hours at 70°C under nitrogen atmosphere. The reaction mixture was cooled to room temperature. The reaction was quenched by the addition of water (100.0 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3x200 mL). The combined organic layers were washed with saturated brine (100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under vacuum. The residue was applied onto a silica gel column and eluted with petroleum ether / ethyl acetate (0% to 30% ethyl acetate) to afford tert-butyl 4-{3-cyano-9-ethyl- 6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol-8-yl}piperazine-1-carboxylate (11.0 g, 22.0 mmol, 90% purity, 95% yield) as a yellow solid.

[0254] Step-2. Synthesis of 9-ethyl-6,6-dimethyl-11-oxo-8-(piperazin-1-yl)-6,11-dihydro- 5H-benzo[b]carbazole-3-carbonitrile, (INT-6)

[0255] A solution of tert-butyl 4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H- benzo[b]carbazol-8-yl}piperazine-1-carboxylate (100 mg, 200 µmol) in DCM (1 mL) and TFA (1 mL) at room temperature and then stirred for 3.0 hours. The resulting mixture was concentrated in vacuum. The residue was purified by prep-HPLC (NH3·H2O buffer) to afford 9-ethyl-6,6- dimethyl-11-oxo-8-(piperazin-1-yl)-5H,6H,11H-benzo[b]carb azole-3-carbonitrile (29.7 mg, 74.7 µmol) as a white solid (INT-6). Yield: 29.7 mg, 37.2%; Appearance: White solid;1H NMR (300 MHz, DMSO-d6) δ 8.32 (d, J = 8.2 Hz, 1H), 8.08 – 7.97 (m, 2H), 7.60 (dd, J = 8.2, 1.4 Hz, 1H), 7.33 (s, 1H), 2.90 (s, 8H), 2.72 (d, J = 7.5 Hz, 2H), 1.76 (bs, 6H), 1.27 (t, J = 7.0, 6.5 Hz, 3H). HPLC purity: 99.3%; LCMS Calculated for C25H26N4O: 398.21; Observed: 399.3[M+H]+. INT-7. Synthesis of (R)-8-(4-(4-(3-aminopyrrolidin-1-yl)butanoyl)piperazin-1-yl)-9-ethyl-6,6- dimethyl-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrile.

[0256] Step-1. Synthesis of methyl 4-[(3R)-3-{[(tert-butoxy)carbonyl]amino}pyrrolidin-1- yl]butanoate

[0257] To a solution of methyl 4-bromobutanoate (193 mg, 1.07 mmol) in DMF (5 mL) was added methyl 4-bromobutanoate (193 mg, 1.07 mmol), K2CO3(147 mg, 1.07 mmol) and NaI (160 mg, 1.07 mmol) , the mixture reaction was stirred at 80 °C for 3 h. The reaction was added to water (60 mL) and extracted with EA (60 mL x 3). The organic layer was washed with water (40 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated to give residue, which was purified by flash silica gel chromatography (PE / EA = 1 / 1) to give product methyl 4-[(3R)-3-{[(tert- butoxy)carbonyl]amino}pyrrolidin-1-yl]butanoate (250 mg, 872 µmol, 90% purity, 82% yield) as a colorless oil

[0258] Step-2. Synthesis of 4-[(3R)-3-{[(tert-butoxy)carbonyl]amino}pyrrolidin-1- yl]butanoic acid

[0259] To a solution of methyl 4-[(3R)-3-{[(tert-butoxy)carbonyl]amino}pyrrolidin-1- yl]butanoate (300 mg, 1.04 mmol) in THF (3 mL) / MeOH (1 mL) / H2O (1 mL) was added lithiumol hydrate (218 mg, 5.20 mmol), the reaction mixture was stirred for 2 h at rt. The reaction mixture was concentrated to give 4-[(3R)-3-{[(tert-butoxy) carbonyl]amino}pyrrolidin-1- yl]butanoic acid (283 mg, 1.04 mmol, 90% purity, 100% yield) as a yellow oil.

[0260] Step-3. Synthesis of 9-ethyl-6,6-dimethyl-11-oxo-8-(piperazin-1-yl)-5H,6H,11H- benzo[b]carbazole-3-carbonitrile

[0261] To a solution of tert-butyl 4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H- benzo[b] carbazol-8-yl}piperazine-1-carboxylate (440 mg, 882 µmol) in DCM (5 mL ) was added TFA (71.4 µL, 882 µmol). The reaction mixture was stirred at rt for 1 h. The reaction mixture was concentrated to give 9-ethyl-6,6-dimethyl-11-oxo-8-(piperazin-1-yl)-5H,6H,11H- benzo[b]carbazole-3-carbonitrile (330 mg, 828 µmol, 93% purity, 94% yield) as a yellow oil.

[0262] Step-4. Synthesis of tert-butyl N-[(3R)-1-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4- oxobutyl]pyrrolidin-3-yl]carbamate

[0263] To a solution of 9-ethyl-6,6-dimethyl-11-oxo-8-(piperazin-1-yl)-5H,6H,11H- benzo[b]carbazole-3-carbonitrile (350 mg, 878 µmol) in DMF (10 mL ) was added 4-[(3R)-3- {[(tert-butoxy)carbonyl]amino}pyrrolidin-1-yl]butanoic acid (285 mg, 1.05 mmol), DIPEA (723 µL, 4.39 mmol) and HATU (497 mg, 1.31 mmol) , the mixture reaction was stirred at rt for 4 h. The reaction was added water (200 mL) and extracted with EA (300 mL x 3). The organic layer was washed with water (200 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated to give residue, which was purified by flash silica gel chromatography (DCM / MeOH = 10 / 1) to give product tert-butyl N-[(3R)-1-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H- benzo[b]carbazol-8-yl}piperazin-1-yl)-4- oxobutyl]pyrrolidin-3-yl]carbamate (450 mg, 689 µmol, 100% purity, 79% yield) as a yellow solid.

[0264] Step-5. Synthesis of 8-(4-{4-[(3R)-3-aminopyrrolidin-1-yl]butanoyl}piperazin-1-yl)- 9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazole-3-carbonitrile

[0265] To a solution of tert-butyl N-[(3R)-1-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4- oxobutyl]pyrrolidin-3-yl]carbamate (400 mg, 612 µmol) in DCM (5 mL ) was added TFA (1.5 mL, 612 µmol) , the reaction mixture was stirred at rt for 1 h. The reaction mixture was concentrated to give 8-(4-{4-[(3R)-3- aminopyrrolidin-1-yl]butanoyl}piperazin-1-yl)-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H- benzo[b]carbazole-3-carbonitrile INT-7 (300 mg, 542 µmol, 94% purity, 89% yield) as a yellow oil. LCMS Calculated for C33H40N6O2: 552.32; Observed: 553.2 [M+H]+.INT-8. Synthesis of 4-((R)-3-((S)-2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H-imidazol-1- yl)pentanamido)pyrrolidin-1-yl)butanoic acid.

[0266] Step-1. Synthesis of ethyl (R)-4-(3-((tert-butoxycarbonyl)amino)pyrrolidin-1- yl)butanoate

[0267] To a solution of tert-butyl N-[(3R)-pyrrolidin-3-yl]carbamate (1g, 5.36 mmol) in DMF (10 mL) was added ethyl 4-bromobutanoate (1.25 g, 6.43 mmol) , K2CO3(2.20 g, 16.0 mmol) at rt. The reaction mixture was stirred for 2 h at 80 °C. The reaction mixture was added water (20 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (MeOH / DCM = 0-10%) to give ethyl 4-[(3R)-3-{[(tert-butoxy)carbonyl]amino}pyrrolidin-1-yl]butanoate (1.26 g, 4.19 mmol, 100% purity, 78% yield) as a light yellow oil.

[0268] Step-2. Synthesis of ethyl (R)-4-(3-aminopyrrolidin-1-yl)butanoate

[0269] To a solution of ethyl 4-[(3R)-3-{[(tert-butoxy)carbonyl]amino}pyrrolidin-1- yl]butanoate (160mg, 532 µmol) in DCM (4 mL) was added TFA (1 ml, 4.19 mmol) at rt. The reaction mixture was stirred for 2 h at rt. The reaction mixture was concentrated under reduced pressure to give ethyl 4-[(3R)-3-aminopyrrolidin-1-yl]butanoate (106 mg, 529 µmol, 100% purity, 100% yield ) as a light yellow oil.

[0270] Step-3. Synthesis of ethyl 4-((R)-3-((S)-2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H- imidazol-1-yl)pentanamido)pyrrolidin-1-yl)butanoate

[0271] To a solution of ethyl 4-[(3R)-3-aminopyrrolidin-1-yl]butanoate (106mg, 529 µmol) in DMF (5 mL) was added (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H-imidazol-1- yl)pentanoic acid (190 mg, 581 µmol) , DIPEA (343 mg, 2.64 mmol) , DEPBT (237 mg, 793µmol) at rt. The reaction mixture was stirred for 2 h at rt. The reaction mixture was added water (20 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (MeOH / DCM = 0-10%) to give ethyl 4-[(3R)-3-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H-imidazol-1- yl)pentanamido] pyrrolidin-1-yl]butanoate (170 mg, 332 µmol, 100% purity, 62% yield) as a light yellow oil.

[0272] Step-4. Synthesis of 4-((R)-3-((S)-2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H- imidazol-1-yl)pentanamido)pyrrolidin-1-yl)butanoic acid

[0273] To a solution of ethyl 4-[(3R)-3-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro- 1H-imidazol-1-yl)pentanamido]pyrrolidin-1-yl]butanoate (170mg, 332 µmol) in THF (5 mL) and H2O (1 mL) was added LiOH (39.8 mg, 1.66 mmol) at rt. The reaction mixture was stirred for 2 h at rt. The reaction mixture was adjust pH to 5-6 with 1 N HCl (aq.) and then extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (MeOH / DCM = 0-30%) to give 4-[(3R)-3-[(2S)-2-{[(tert- butoxy)carbonyl]amino}-5-(2-nitro-1H-imidazol-1-yl)pentanamido]pyrrolidin-1-yl]butanoic acid, INT-8 (120 mg, 248 µmol, 100% purity, 75% yield) as a light yellow oil. LCMS Calculated for C21H34N6O7: 482.25; Observed: 483.1 [M+H]+. INT-10. Synthesis of 8-(4-{4-[(4R)-4-amino-3,3-dimethylpyrrolidin-1-yl]butanoyl}piperazin-1- yl)-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazole-3-carbonitrile

[0274] Step-1. Synthesis of methyl 4-[(4R)-4-{[(tert-butoxy)carbonyl]amino}-3,3- dimethylpyrrolidin-1-yl]butanoate

[0275] To a solution of tert-butyl N-[(3R)-4,4-dimethylpyrrolidin-3-yl]carbamate (250 mg, 1.16 mmol) and methyl 4-bromobutanoate (376 mg, 2.08 mmol) in MeCN (5 mL) was added DIPEA (599 mg, 4.64 mmol), the mixture was stirred at 80 °C for 2 h. After the reaction was completed, the mixture was concentrated under reduced pressure to give the crude residue, which was purified by flash silica gel chromatography (DCM / MeOH = 20 / 1) to give the product methyl 4-[(4R)-4-{[(tert-butoxy)carbonyl]amino}-3,3-dimethylpyrrolidin-1-yl]butanoate (300 mg, 954 µmol) as a yellow oil.

[0276] Step-2. Synthesis of 4-[(4R)-4-{[(tert-butoxy)carbonyl]amino}-3,3- dimethylpyrrolidin-1-yl]butanoic acid

[0277] To a solution of methyl 4-[(4R)-4-{[(tert-butoxy)carbonyl]amino}-3,3- dimethylpyrrolidin-1-yl]butanoate (300 mg, 954 µmol) in THF (5 mL) and water (2.5 mL) was added LiOH (45.6 mg, 1.9 mmol), the mixture was stirred at rt for 2 h. After the reaction was completed, the mixture was concentrated under reduced pressure to give the crude product 4-[(4R)- 4-{[(tert-butoxy)carbonyl]amino}-3,3-dimethylpyrrolidin-1-yl]butanoic acid (279 mg, 927 µmol) which was directly used in the next steps.

[0278] Step-3. Synthesis of tert-butyl N-[(3R)-1-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]-4,4-dimethylpyrrolidin-3- yl]carbamate

[0279] To a solution of 4-[(4R)-4-{[(tert-butoxy)carbonyl]amino}-3,3-dimethylpyrrolidin-1- yl]butanoic acid (200 mg, 665 µmol), DIPEA (429 mg, 3.32 mmol) and HATU (379 mg, 997 µmol) in DMF (5 mL) was added 9-ethyl-6,6-dimethyl-11-oxo-8-(piperazin-1-yl)-5H,6H,11H- benzo[b]carbazole-3-carbonitrile (265 mg, 665 µmol), the mixture was stirred at rt for 16 h. After the reaction was completed, the mixture was concentrated under reduced pressure to give the crude residue, which was purified by prep-HPLC(ACN / water / 0.1% FA) to give the product tert-butyl N- [(3R)-1-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol-8- yl}piperazin-1-yl)-4-oxobutyl]-4,4-dimethylpyrrolidin-3-yl]carbamate (263 mg, 386 µmol) as a yellow solid.

[0280] Step-4. Synthesis of 8-(4-{4-[(4R)-4-amino-3,3-dimethylpyrrolidin-1- yl]butanoyl}piperazin-1-yl)-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazole-3- carbonitrile

[0281] To a solution of tert-butyl N-[(3R)-1-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]-4,4-dimethylpyrrolidin-3- yl]carbamate (70 mg, 102 µmol) in DCM (3 mL) was added TFA (1.5 mL), the mixture was stirred at rt for 40 min. After the mixture was completed, the mixture was concentrated under reduced pessure to give the crude residue, which was purified by prep-HPLC(ACN / water / 0.1% NH4HCO3) to give the product 8-(4-{4-[(4R)-4-amino-3,3-dimethylpyrrolidin-1-yl]butanoyl}piperazin-1-yl)- 9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazole-3-carbonitrile, (INT-10). Yield: 15 mg, 25%; Appearance: White solid;1H NMR (400 MHz, DMSO-d6) δ 8.31 (d, J = 8.4 Hz, 1H), 8.07 (s, 1H), 8.00 (s, 1H), 7.60 (dd, J = 8.0, 1.2 Hz, 1H), 7.39 (s, 1H), 3.65 (s, 4H), 3.01 – 2.79 (m, 6H), 2.75 (q, J = 7.6 Hz, 2H), 2.46 – 2.07 (m, 7H), 1.75 (s, 6H), 1.70 – 1.55 (m, 2H), 1.29 (t, J = 7.6 Hz, 3H), 1.08 – 0.69 (m, 6H); HPLC purity: 92.64%; LCMS Calculated for C35H44N6O2: 580.78; Observed: 581.1 [M+H]+. INT-11. Synthesis of (R)-8-(4-(4-(7-amino-5-azaspiro[2.4]heptan-5-yl)butanoyl)piperazin-1- yl)-9-ethyl-6,6-dimethyl-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrile.

[0282] Step-1. Synthesis of methyl 4-[(7R)-7-{[(tert-butoxy)carbonyl]amino}-5- azaspiro[2.4]heptan-5-yl]butanoate

[0283] A solution of methyl 4-bromobutanoate (300 mg, 1.65 mmol), DIEA (637 mg, 4.94 mmol) and tert-butyl N-[(7R)-5-azaspiro[2.4]heptan-7-yl]carbamate (354 mg, 1.65 mmol) in MeCN (5 mL) was stirred for 2 hours at 80°C. The resulting mixture was concentrated in vacuum to afford methyl 4-[(7R)-7-{[(tert-butoxy)carbonyl]amino}-5-azaspiro[2.4]heptan-5-yl]butanoate(301 mg, 965 µmol, 86.0% purity, 59.0% yield) as a yellow oil, which was used for next step directly.

[0284] Step-2. Synthesis of 4-[(7R)-7-{[(tert-butoxy)carbonyl]amino}-5-azaspiro[2.4]heptan- 5-yl] butanoic acid

[0285] A solution of methyl 4-[(7R)-7-{[(tert-butoxy)carbonyl]amino}-5- azaspiro[2.4]heptan-5-yl]butanoate (350 mg, 1.12 mmol) and NaOH (134 mg, 3.36 mmol) in MeOH / H2O (1:1, 10 mL) was stirred for 2 hours at room temperature. The resulting mixture was concentrated in vacuum to afford crude 4-[(7R)-7-{[(tert-butoxy)carbonyl]amino}-5- azaspiro[2.4]heptan-5-yl]butanoic acid (256 mg, 860 µmol, 86.0% purity, 77.0% yield) as a yellow oil, which was used for next step directly.

[0286] Step-3. Synthesis of tert-butyl N-[(7R)-5-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11- oxo-5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]-5-azaspiro[2.4]heptan-7-yl] carbamate

[0287] A mixture of 4-[(7R)-7-{[(tert-butoxy)carbonyl]amino}-5-azaspiro[2.4]heptan-5- yl]butanoic acid (300 mg, 1.00 mmol), DIEA (398 mg, 1.00 mmol) and 9-ethyl-6,6-dimethyl-11- oxo-8-(piperazin-1-yl)-5H,6H,11H-benzo[b]carbazole-3-carbonitrile (398 mg, 1.00 mmol) in DMF (5 mL) was stirred at room temperature for 10 min. HATU (455 mg, 1.20 mmol) was then added, and the reaction mixture was stirred for 1.0 hour at room temperature. The resulting mixture was concentrated in vacuum, the residue was purified by prep-HPLC (NH3·H2O buffer) to give tert-butyl N-[(7R)-5-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol- 8-yl}piperazin-1-yl)-4-oxobutyl]-5-azaspiro[2.4]heptan-7-yl]carbamate (346 mg, 509 µmol, 87.0% purity, 51.0% yield) as a yellow solid.

[0288] Step-4. Synthesis of 8-(4-{4-[(7R)-7-amino-5-azaspiro[2.4]heptan-5- yl]butanoyl}piperazin-1-yl)-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazole-3- carbonitrile, INT-11.

[0289] A solution of tert-butyl N-[(7R)-5-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]-5-azaspiro[2.4]heptan-7- yl]carbamate (180 mg, 265 µmol) in TFA / DCM (1:1, 5 mL) was stirred for 1 hour at room temperature. The resulting mixture was concentrated in vacuum to afford 8-(4-{4-[(7R)-7-amino- 5-azaspiro[2.4]heptan-5-yl]butanoyl}piperazin-1-yl)-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazole-3-carbonitrile INT-11 (127 mg, 220 µmol, 85% purity, 83% yield) as a brown oil, which was used for next step directly. INT-12. Synthesis of (2S)-5-{2-amino-5-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]pyridin-3-yl}-2-{[(tert- butoxy)carbonyl]amino}pentanoic acid

[0290] Step-1. Synthesis of methyl (2S)-5-(2-amino-5-bromopyridin-3-yl)-2-{[(tert- butoxy)carbonyl]amino}pent-4-ynoate

[0291] To a solution of methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}pent-4-ynoate (250 mg, 1.10 mmol), 5-bromo-3-iodopyridin-2-amine (273 mg, 916 µmol), Pd(PPh3)2Cl2(128 mg, 183 µmol) and CuI (17.4 mg, 91.6 µmol) in DMF (5 mL) was added TEA (463 mg, 4.58 mmol) at rt. The reaction mixture was stirred for 4 h at 80 °C. The reaction mixture was added water (20 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (MeOH / DCM = 0-5%) to give methyl (2S)-5-(2-amino-5-bromopyridin-3-yl)-2-{[(tert-butoxy)carbonyl]amino}pent-4-ynoate (258 mg, 647 µmol, 100% purity, 71% yield) as a yellow solid.

[0292] Step-2. Synthesis of (3E)-4-{6-amino-5-[(4S)-4-{[(tert-butoxy)carbonyl]amino}-5- methoxy-5-oxopent-1-yn-1-yl]pyridin-3-yl}but-3-enoic acid

[0293] To a solution of methyl (2S)-5-(2-amino-5-bromopyridin-3-yl)-2-{[(tert-butoxy) carbonyl]amino}pent-4-ynoate (290 mg, 728 µmol), but-3-enoic acid (155 mg, 1.81 mmol),(acetyloxy)palladio acetate (24.4 mg, 109 µmol), P(o-tol)3(66.3 mg, 218 µmol) in CH3CN (5 mL) was added TEA (367 mg, 3.63 mmol) at rt. The reaction mixture was stirred for 0.5 h at 120 °C under microwave irradition. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (MeOH / DCM = 0-10%) to give (3E)-4-{6-amino-5-[(4S)-4-{[(tert-butoxy)carbonyl]amino}-5-methoxy-5-oxopent-1-yn-1- yl]pyridin-3-yl}but-3-enoic acid (185 mg, 458 µmol, 92% purity, 63% yield) as a brown solid.

[0294] Step-3. Synthesis of 4-{6-amino-5-[(4S)-4-{[(tert-butoxy)carbonyl]amino}-5- methoxy-5-oxopentyl]pyridin-3-yl}butanoic acid

[0295] To a solution of (3E)-4-{6-amino-5-[(4S)-4-{[(tert-butoxy)carbonyl]amino}-5- methoxy-5-oxopent-1-yn-1-yl]pyridin-3-yl}but-3-enoic acid (185 mg, 458 µmol) in MeOH (5 mL) was added Pd / C (97.3 mg, 91.6 µmol) at rt. The reaction mixture was stirred for 16 h at rt under H2. The mixture was filtered through a celite pad, and the filtrate was concentrated under reduced pressure to give 4-{6-amino-5-[(4S)-4-{[(tert-butoxy)carbonyl]amino}-5-methoxy-5- oxopentyl]pyridin-3-yl}butanoic acid (155 mg, 378 µmol, 85% purity, 83% yield) as a light yellow solid.

[0296] Step-4. Synthesis of methyl (2S)-5-{2-amino-5-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl- 11-oxo-5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]pyridin-3-yl}-2-{[(tert- butoxy)carbonyl]amino}pentanoate

[0297] To a solution of 4-{6-amino-5-[(4S)-4-{[(tert-butoxy)carbonyl]amino}-5-methoxy-5- oxopentyl]pyridin-3-yl}butanoic acid (155 mg, 378 µmol) and 9-ethyl-6,6-dimethyl-11-oxo-8- (piperazin-1-yl)-5H,6H,11H-benzo[b]carbazole-3-carbonitrile (150 mg, 378 µmol) in DCM (5 mL) was added DIPEA (242 mg, 1.88 mmol) and HATU (172 mg, 453 µmol) at rt. The reaction mixture was stirred for 2 h at rt. The reaction mixture was added water (20 mL) and extracted with DCM (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (MeOH / DCM = 0-10%) to give methyl (2S)-5-{2-amino-5-[4- (4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4- oxobutyl]pyridin-3-yl}-2-{[(tert-butoxy)carbonyl]amino}pentanoate (103 mg, 130 µmol, 91% purity, 35% yield) as a yellow solid.

[0298] Step-5. Synthesis of (2S)-5-{2-amino-5-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]pyridin-3-yl}-2-{[(tert-butoxy) carbonyl]amino}pentanoic acid

[0299] To a solution of methyl (2S)-5-{2-amino-5-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11- oxo-5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]pyridin-3-yl}-2-{[(tert- butoxy) carbonyl]amino}pentanoate (103 mg, 130 µmol) in THF (5 mL) and H2O (2 mL) was added LiOH·H2O (27.2 mg, 650 µmol) at rt. The reaction mixture was stirred for 2 h at rt. The reaction mixture was concentrated under reduced pressure to give (2S)-5-{2-amino-5-[4-(4-{3- cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4- oxobutyl] pyridin-3-yl}-2-{[(tert-butoxy)carbonyl]amino}pentanoic acid (100 mg, 128 µmol, 91% purity, 100% yield) as a Li salt, which was used for the next step without further purification. INT-13. Synthesis of (S)-5-(2-amino-4-(5-(4-(3-cyano-9-ethyl-6,6-dimethyl-11-oxo-6,11- dihydro-5H-benzo[b]carbazol-8-yl)piperazin-1-yl)-5-oxopentyl)-1H-imidazol-1-yl)-2-((tert- butoxycarbonyl)amino)pentanoic acid.

[0300] Step-1. Synthesis of tert-butyl (S)-2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H- imidazol-1-yl)pentanoate

[0301] A mixture of tert-butyl (2S)-5-bromo-2-{[(tert-butoxy)carbonyl]amino}pentanoate (5.00 g, 14.1 mmol), K2CO3(5.83 g, 42.3 mmol), 2-nitro-1H-imidazole (1.75 g, 15.5 mmol) and DMF (50 mL) at room temperature was stirred for 12 hours at 50 ° C. The reaction mixture was cooled to room temperature and quenched by the addition of water (100.0 mL). The resultingmixture was extracted with ethyl acetate (3x200 mL). The combined organic layers were washed with saturated salt water (3x100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated in vacuum to afford tert-butyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2- nitro-1H-imidazol-1-yl)pentanoate (4.50 g, 11.7 mmol, 90% purity, 82% yield) as a yellow solid.

[0302] Step-2. Synthesis of tert-butyl (2S)-5-(4-bromo-2-nitro-1H-imidazol-1-yl)-2-{[(tert- butoxy)carbonyl]amino}pentanoate

[0303] A mixture of tert-butyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H- imidazol-1-yl)pentanoate (4.00 g, 10.4 mmol), NBS (2.77g, 15.6 mmol) and DMF (50 mL) at room temperature was stirred for 12 hours at 50 ° C. The reaction mixture was cooled to room temperature and quenched by the addition of water (100.0 mL). The resulting mixture was extracted with ethyl acetate (3x100 mL). The combined organic layers were washed with saturated salt water (3x100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated in vacuum to afford tert-butyl (2S)-5-(4-bromo-2-nitro-1H-imidazol-1-yl)-2-{[(tert- butoxy)carbonyl]amino}pentanoate (3.00 g, 6.47 mmol, 90% purity, 65% yield) as a yellow solid.

[0304] Step-3. Synthesis of methyl (4E)-5-{1-[(4S)-5-(tert-butoxy)-4-{[(tert- butoxy)carbonyl]amino}-5-oxopentyl]-2-nitro-1H-imidazol-4-yl}pent-4-enoate

[0305] To a solution of tert-butyl (2S)-5-(4-bromo-2-nitro-1H-imidazol-1-yl)-2-{[(tert- butoxy)carbonyl]amino}pentanoate (2.00 g, 4.31 mmol) and methyl pent-4-enoate (1.47 g, 12.9 mmol) in DMF (20 mL) was added TEA (2.17 g, 21.5 mmol) and Pd2(dba)3(788 mg, 862 µmol), P(O-Tol)3(1.31 g, 4.31 mmol). The resulting mixture was stirred for 12 hours at 100 ° C under nitrogen atmosphere. The mixture was allowed to cool down to room temperature and concentrated in vacuum. The residue was purified by reverse phase flash with the following conditions (column, C18 silica gel; mobile phase, H2O / 0.1% NH4OH in MeCN, 10% to 50% gradient in 10 min; detector, UV 254 nm) to afford (4E)-5-{1-[(4S)-5-(tert-butoxy)-4-{[(tert- butoxy)carbonyl]amino}-5-oxopentyl]-2-nitro-1H-imidazol-4-yl}pent-4-enoate (1.00 g, 2.01 mmol, 90% purity, 47% yield) as a yellow oil.

[0306] Step-4. Synthesis of (4E)-5-{1-[(4S)-5-(tert-butoxy)-4-{[(tert- butoxy)carbonyl]amino}-5-oxopentyl]-2-nitro-1H-imidazol-4-yl}pent-4-enoic acid

[0307] To a stirred solution of methyl (4E)-5-{1-[(4S)-5-(tert-butoxy)-4-{[(tert- butoxy)carbonyl]amino}-5-oxopentyl]-2-nitro-1H-imidazol-4-yl}pent-4-enoate (500 mg, 1.00 mmol) in MeOH (5 mL) and H2O (2 mL) was added LiOH (205 mg, 5.00 mmol) at roomtemperature. The resulting mixture was stirred for 2 hours at room temperature. The mixture was concentrated in vacuum to afford the crude product (4E)-5-{1-[(4S)-5-(tert-butoxy)-4-{[(tert- butoxy)carbonyl]amino}-5-oxopentyl]-2-nitro-1H-imidazol-4-yl}pent-4-enoic acid (300 mg, 621 µmol, 90% purity, 62% yield) as a yellow solid.

[0308] Step-5. Synthesis of (2S)-5-(4-bromo-2-nitro-1H-imidazol-1-yl)-2-{[(tert- butoxy)carbonyl]amino}pentanoate

[0309] To a solution of 9-ethyl-6,6-dimethyl-11-oxo-8-(piperazin-1-yl)-5H,6H,11H- benzo[b]carbazole-3-carbonitrile (450 mg, 1.13 mmol) in DMF (10 mL) were added K2CO3(426 mg, 3.09 mmol) at room temperature, stirring for 15 min at room temperature. To above mixture was added (4E)-5-{1-[(4S)-5-(tert-butoxy)-4-{[(tert-butoxy)carbonyl]amino}-5-oxopentyl]-2- nitro-1H-imidazol-4-yl}pent-4-enoic acid (500 mg, 1.03 mmol) and HATU (585 mg, 1.54 mmol) at room temperature. The resulting mixture was stirred for 1 hour at room temperature. The reaction was quenched by the addition of water (200 mL) at room temperature. The precipitated solids were collected by filtration and washed with water (3x100 mL). The resulting solid was dried under infrared light. This resulted in tert-butyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{4- [(1E)-5-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol-8-yl}piperazin- 1-yl)-5-oxopent-1-en-1-yl]-2-nitro-1H-imidazol-1-yl}pentanoate (400 mg, 463 µmol, 90% purity, 50% yield) as a yellow solid.

[0310] Step-6. Synthesis of (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{4-[(1E)-5-(4-{3- cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-5- oxopent-1-en-1-yl]-2-nitro-1H-imidazol-1-yl}pentanoic acid.

[0311] To a stirred solution of tert-butyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{4-[(1E)-5- (4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol-8-yl} piperazin-1-yl)-5- oxopent-1-en-1-yl]-2-nitro-1H-imidazol-1-yl}pentanoate (400 mg, 463 µmol) in MeOH (10 mL) and H2O (5 mL) was added NaOH (18.5 mg, 463 µmol) at room temperature. The resulting mixture was stirred for 1 hour at 50 °C. The reaction mixture was cooled to room temperature and concentrated in vacuum. The residue was purified by reverse flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water, 30% to 55% gradient in 10 min; detector, UV 220 nm. This resulted in (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{4- [(1E)-5-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol-8-yl}piperazin- 1-yl)-5-oxopent-1-en-1-yl]-2-nitro-1H-imidazol-1-yl}pentanoic acid, INT-13. Yield: 300 mg,90%; Appearance: Yellow solid;1H NMR (300 MHz, DMSO-d6) δ 8.22 (d, J = 3.2 Hz, 1H), 8.05 (s, 1H), 7.96 – 7.91 (m, 2H), 7.68 (s, 1H), 7.47 – 7.30 (m, 2H), 6.46 – 6.28 (m, 1H), 5.99 – 5.97 (m, 1H), 4.29 – 4.28 (m, 1H), 3.69 – 3.67 (m, 4H), 2.92 – 2.86 (m, 6H), 2.82 – 2.69 (m, 4H), 2.57 (d, J = 6.4 Hz, 2H), 1.79 – 1.60 (m, 10H), 1.36 (s, 9H), 1.29 (t, J = 5.4 Hz, 3H). HPLC purity: 90%; LCMS Calculated for C43H50N8O8:806.38; Observed: 807.5 [M+H]+. INT-14. Synthesis of tert-butyl N-[(1S)-4-amino-1-{[(3R)-1-ethylpyrrolidin-3- yl]carbamoyl}butyl]carbamate.

[0312] Step-1. Synthesis of tert-butyl N-[(3R)-1-ethylpyrrolidin-3-yl]carbamate

[0313] To a stirred solution of tert-butyl N-[(3R)-pyrrolidin-3-yl]carbamate (4.00 g, 21.4 mmol) and bromoethane (2.78 g, 25.6 mmol) in MeCN (50 mL) was added K2CO3(8.84 g, 64.1 mmol) at room temperature. The resulting mixture was stirred for 16 hours at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 150 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated in vacuum. This resulted in tert-butyl N-[(3R)-1-ethylpyrrolidin-3-yl]carbamate (2.40 g, 11.1 mmol, 85% purity, 53% yield) as a yellow solid.

[0314] Step-2. Synthesis of (R)-1-ethylpyrrolidin-3-amine hydrochloride

[0315] A solution of tert-butyl N-[(3R)-1-ethylpyrrolidin-3-yl]carbamate (2.40 g, 11.1 mmol) in HCl / dioxane (4M, 20 mL) was stirred for 1 hour at room temperature. The resulting mixture was concentrated in vacuum to afford (R)-1-ethylpyrrolidin-3-amine as HCl salt (1.60 g, 10.6 mmol, 80% purity, crude) as a brown solid, which was used for next step directly.

[0316] Step-3. Synthesis of tert-butyl N-[(1S)-4-{[(benzyloxy)carbonyl]amino}-1-{[(3R)-1- ethylpyrrolidin-3-yl]carbamoyl}butyl]carbamate

[0317] A mixture of (2S)-5-{[(benzyloxy)carbonyl]amino}-2-{[(tert- butoxy)carbonyl]amino}pentanoic acid (2.00 g, 5.45 mmol), (3R)-1-ethylpyrrolidin-3-amine hydrochloride (985 mg, 6.54 mmol) and NMI (1.55 g, 19.0 mmol) in MeCN (20 mL) was stirred at 0 for 10 min. TCFH (1.83 g, 6.54 mmol) was then added, and the reaction mixture was stirred for 3.0 hours at room temperature. The reaction was diluted with water (100 mL) and extracted with ethyl acetate (100 mLx3). The combined organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuum. The residue was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate (10% to 40% ethyl acetate) to afford tert-butyl N-[(1S)-4-{[(benzyloxy)carbonyl]amino}-1-{[(3R)-1- ethylpyrrolidin-3-yl]carbamoyl} butyl]carbamate (1.80 g, 3.89 mmol, 95% purity, 71% yield) as a white solid.

[0318] Step-4. Synthesis of tert-butyl N-[(1S)-4-amino-1-{[(3R)-1-ethylpyrrolidin-3- yl]carbamoyl}butyl]carbamate, (INT-14)

[0319] Into a 50 mL round-bottom flask, was placed tert-butyl N-[(1S)-4- {[(benzyloxy)carbonyl]amino}-1-{[(3R)-1-ethylpyrrolidin-3-yl]carbamoyl}butyl]carbamate (1.8 g, 3.89 mmol) in MeOH (20 mL) was added Pd / C (372 mg, 3.50 mmol, 10%) at room temperature under nitrogen atmosphere and then the reaction mixture was degassed and purged with hydrogen for three times. The resulting reaction mixture was stirred for 3 hours under hydrogen atmosphere (1atm pressure). The reaction mixture was filtered and the filtrate was concentrated in vacuum to give tert-butyl N-[(1S)-4-amino-1-{[(3R)-1-ethylpyrrolidin-3-yl]carbamoyl}butyl]carbamate as a white solid, (INT-14). Yield: 1.20 g, 94%;1H NMR (300 MHz, DMSO-d6) δ 7.92 (d, J = 7.40 Hz, 1H), 6.74 (d, J = 8.5 Hz, 1H), 4.19 ̽ 4.03 (m, 1H), 3.93 ̽ 3.81 (m, 1H), 2.95 ̽ 2.83 (m, 1H), 2.67 ̽ 2.53 (m, 4H), 2.45 ̽ 2.24 (m, 4H), 2.15 ̽ 1.98 (m, 1H), 1.66 ̽ 1.42 (m, 4H), 1.38 (s, 9H), 1.01 (t, J = 7.2 Hz, 3H). HPLC purity: 90%; LCMS Calculated for: C16H32N4O3:328.25; Observed: 329.1 [M+H]+. INT-15. Synthesis of 8-[4-(4-aminobutanoyl)piperazin-1-yl]-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazole-3-carbonitrile.

[0320] Step-1. Synthesis of tert-butyl N-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]carbamate

[0321] A mixture of 9-ethyl-6,6-dimethyl-11-oxo-8-(piperazin-1-yl)-5H,6H,11H- benzo[b]carbazole-3-carbonitrile (500 mg, 1.25 mmol), 4-{[(tert- butoxy)carbonyl]amino}butanoic acid (254 mg, 1.25 mmol) and DIEA (483 mg, 3.75 mmol) in DMF (10 mL) was stirred at room temperature for 10 min. HATU (570 mg, 1.50 mmol) was then added, and the reaction mixture was stirred for 1 hour at room temperature. The resulting mixture was concentrated in vacuum, the residue was purified by prep-HPLC (NH3·H2O buffer) to give tert-butyl N-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H-benzo[b]carbazol-8- yl}piperazin-1-yl)-4-oxobutyl]carbamate (690 mg, 1.18 mmol, 98.0% purity, 95.0% yield) as a white solid.

[0322] Step-2. Synthesis of 8-[4-(4-aminobutanoyl)piperazin-1-yl]-9-ethyl-6,6-dimethyl-11- oxo-5H,6H,11H-benzo[b]carbazole-3-carbonitrile, (INT-15)

[0323] A solution of tert-butyl N-[4-(4-{3-cyano-9-ethyl-6,6-dimethyl-11-oxo-5H,6H,11H- benzo[b]carbazol-8-yl}piperazin-1-yl)-4-oxobutyl]carbamate (700 mg, 1.19 mmol) in TFA / DCM (1:1, 10 mL) was stirred for 1 hour at room temperature. The resulting mixture was concentrated in vacuum to afford 8-[4-(4-aminobutanoyl)piperazin-1-yl]-9-ethyl-6,6-dimethyl-11-oxo- 5H,6H,11H-benzo[b]carbazole-3-carbonitrile as TFA salt, (INT-15). Yield: 600 mg, crude;1H NMR (300 MHz, DMSO-d6) δ 12.75 (s, 1H), 8.33 (d, J = 8.1 Hz, 1H), 8.08 (s, 1H), 8.01 (s, 1H), 7.62 (d, J =1.4 Hz, 1H), 7.39 (s, 1H), 3.81 – 3.55 (m, 4H), 3.05 – 2.92 (m, 4H), 2.92 – 2.80 (m,2H), 2.80 – 2.70 (m, 2H), 1.82 (t, J = 7.5 Hz, 2H), 1.76 (s, 6H), 1.36 – 1.16 (m, 5H). HPLC purity: 92.4%; LCMS Calculated for C29H33N5O2: 483.26; Observed:484.3[M+H]+. INT-16. Synthesis of (R)-26-amino-55-fluoro-4,7-dimethyl-6-oxo-11H-3-oxa-7-aza-2(3,5)- pyridina-1(4,3)-pyrazola-5(1,2)-benzenacyclooctaphane-15-carbonitrile, (INT-16):

[0324] Step-1. Synthesis of 3-[(1R)-1-(5-fluoro-2-iodophenyl)ethoxy]pyridin-2-amine

[0325] To a stirred solution of (1S)-1-(5-fluoro-2-iodophenyl)ethanol (200 g, 751 mmol, 1.00 equiv) and 2-aminopyridin-3-ol (91.1 g, 827 mmol, 1.10 equiv) and PPh3 (295 g, 1.13 mol, 1.50 equiv) in THF (4.00 L) was added DIAD (228 g, 1.13 mol, 1.50 equiv) dropwise at 0 under N2 atmosphere. The resulting mixture was stirred for 12 hours at room temperature under N2 atmosphere. The reaction was quenched with H2O (100 mL) at 0 . The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with ethyl acetate / petroleum ether (0 to 50% ethyl acetate) to afford 3-[(1R)-1-(5-fluoro-2- iodophenyl)ethoxy]pyridin-2-amine (220 g, 57%) as a yellow oil.

[0326] Step-2. Synthesis of methyl 2-[(1R)-1-[(2-aminopyridin-3-yl)oxy]ethyl]-4- fluorobenzoate

[0327] To a solution of 3-[(1R)-1-(5-fluoro-2-iodophenyl)ethoxy]pyridin-2-amine (220 g, 614 mmol, 1.00 equiv) and TEA (186 g, 1.84 mol, 3.0 equiv) in MeOH (8.00 L) was added Pd(dppf)Cl2(44.9 g, 61.4mmol, 0.1 equiv) in a pressure tank. The mixture was purged with carbon monoxide for three times and then was pressurized to 20.0 Psi with carbon monoxide at 100 for 12 hours.The reaction mixture was cooled to room temperature and filtered to remove insoluble solids. The filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with ethyl acetate / petroleum ether (0 to 50% ethyl acetate) to afford methyl 2-[(1R)-1-[(2-aminopyridin-3-yl)oxy]ethyl]-4-fluorobenzoate (220 g, 86%) as a green oil.

[0328] Step-3. Synthesis of methyl 2-[(1R)-1-[(2-amino-5-bromopyridin-3-yl)oxy]ethyl]-4- fluorobenzoate

[0329] To a stirred solution of methyl 2-[(1R)-1-[(2-aminopyridin-3-yl)oxy]ethyl]-4- fluorobenzoate (120 g, 413 mmol, 1.00 equiv) in MeCN (1.00 L) were added NBS (88.3 g, 496 mmol, 1.20 equiv) dropwise at 0 under N2. The resulting mixture was stirred for 2 hours at 0 - 10 under N2 atmosphere. The resulting mixture was concentrated under vacuum. The residue was dissolved in ethyl acetate (1.00 L). The mixture was basified to pH 8-9 with sat NaHCO3solution. The resulting mixture was extracted with ethyl acetate (3 x 500 mL). The combined organic layers were washed with brine (2x200 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with ethyl acetate / petroleum ether (0 to 30% ethyl acetate) to afford methyl 2-[(1R)-1-[(2-amino-5-bromopyridin-3-yl)oxy]ethyl]-4-fluorobenzoate (60.0 g, 33%) as a yellow solid.

[0330] Step-4. Synthesis of methyl (R)-2-(1-((2-amino-5-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)pyridin-3-yl)oxy)ethyl)-4-fluorobenzoate

[0331] To a solution of methyl 2-[(1R)-1-[(2-amino-5-bromopyridin-3-yl)oxy]ethyl]-4- fluorobenzoate (60.0 g, 163 mmol, 1.00 equiv) and KOAc (39.9 g, 406 mmol, 2.50 equiv), bis(pinacolato)diboron (49.5 g, 195 mmol, 1.20 equiv) in dioxane (600 mL) was added Pd(dppf)Cl2(13.2 g, 16.3 mmol, 0.100 equiv). After stirring for 16 hours at 80 oC under a nitrogen atmosphere. The mixture was allowed to cool down to room temperature and used to next step directly.

[0332] Step-5. Synthesis of methyl 2-[(1R)-1-([2-amino-5-(3-([(tert-butoxycarbonyl)(methyl) amino]methyl-5-cyano-1H-pyrazol-4-yl)pyridin-3-yl]oxyethyl]-4-fluorobenzoate

[0333] To the mixture from previous step was added tert-butyl N-[(5-cyano-4-iodo-1H- pyrazol-3-yl)methyl]-N-methylcarbamate (47.9 g, 132 mmol, 1.10 equiv), H2O (200 mL), K2CO3(49.8 g, 360 mmol, 3.00 equiv) and Pd(DTBPF)Cl2(15.7 g, 24.0 mmol, 0.20 equiv) at room temperature. After stirring for 16 hours at 100 oC under nitrogen atmosphere, the mixture wasallowed to cool down to room temperature and concentrated under reduced pressure. The residue was dissolved in ethyl acetate (500 mL) and H2O (500 mL). The resulting mixture was extracted with ethyl acetate (3 x 500 mL). The combined organic layers were washed with brine (2 x 200 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with ethyl acetate / petroleum ether (0 to 80% ethyl acetate) to afford methyl 2-[(1R)-1-([2-amino-5-(3-([(tert- butoxycarbonyl)(methyl)amino]methyl-5-cyano-1H-pyrazol-4-yl)pyridin-3-yl]oxyethyl]-4- fluorobenzoate (25.0 g, 31 %) as a yellow solid.

[0334] Step-6. Synthesis of methyl 2-[(1R)-1-[(2-amino-5-(3-cyano-5- [(methylamino)methyl]-2H-pyrazol-4-ylpyridin-3-yl)oxy]ethyl]-4-fluorobenzoate

[0335] To a stirred solution of methyl 2-[(1R)-1-([2-amino-5-(3-([(tert- butoxycarbonyl)(methyl)amino] methyl-5-cyano-1H-pyrazol-4-yl)pyridin-3-yl]oxyethyl]-4- fluorobenzoate (25.0 g, 47.7 mmol, 1.00 equiv) in HCl(gas) in 1,4-dioxane (4M, 250 mL). The resulting mixture was stirred for 2 hours at room temperature. The resulting mixture was concentrated under vacuum. The crude product methyl 2-[(1R)-1-[(2-amino-5-(3-cyano-5- [(methylamino)methyl]-2H-pyrazol-4-ylpyridin-3-yl)oxy]ethyl]-4-fluorobenzoate as HCl salt (18.0 g, 71%) as a light yellow solid, which was used in the next step directly without further purification.

[0336] Step-7. Synthesis of 2-[(1R)-1-[(2-amino-5-(3-cyano-5-[(methylamino)methyl]-2H- pyrazol-4-ylpyridin-3-yl)oxy]ethyl]-4-fluorobenzoic acid

[0337] To a stirred solution of methyl 2-[(1R)-1-[(2-amino-5-(3-cyano-5- [(methylamino)methyl]-2H-pyrazol-4-ylpyridin-3-yl)oxy]ethyl]-4-fluorobenzoate (18.0 g, 42.4 mmol, 1.00 equiv) in MeOH (100 mL) and H2O (50 mL) was added NaOH (5.09 g, 127 mmol, 3.00 equiv) in portions at room temperature. The resulting mixture was stirred for 3 hours at 40 . The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under vacuum to give crude product 2-[(1R)-1-[(2-amino-5-(3-cyano-5- [(methylamino)methyl]-2H-pyrazol-4-ylpyridin-3-yl)oxy]ethyl]-4-fluorobenzoic acid (13.0 g, 56%) as light yellow solid, which was used in the next step directly.

[0338] Step-8. Synthesis of (R)-26-amino-55-fluoro-4,7-dimethyl-6-oxo-11H-3-oxa-7-aza- 2(3,5)-pyridina-1(4,3)-pyrazola-5(1,2)-benzenacyclooctaphane-15-carbonitrile, PH-CSM-INT- 10 / CSM-2362073 (INT-16)

[0339] To a stirred solution of 2-[(1R)-1-[(2-amino-5-(3-cyano-5-[(methylamino)methyl]-2H- pyrazol-4-ylpyridin-3-yl)oxy]ethyl]-4-fluorobenzoic acid (13.0 g, 31.7 mmol, 1.00 equiv) and K2CO3(8.76 g, 63.3 mmol, 2.00 equiv) in DMF (200 mL) was added PyBOP (24.7 g, 47.5 mmol, 1.50 equiv) in portions at room temperature. The resulting mixture was stirred for 2 hours at room temperature. The mixture was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in H2O (0.01% TFA), 30% to 55% gradient in 15 min; detector, UV 254 nm. To give (R)-26-amino-55-fluoro-4,7-dimethyl-6-oxo- 11H-3-oxa-7-aza-2(3,5)-pyridina-1(4,3)-pyrazola-5(1,2)-benzenacyclooctaphane-15-carbonitrile, PH-CSM-INT-10 / CSM-2362073 (INT-16). Yield: 6.0 g, 45%. Appearance: white solid; 1H NMR (400 MHz, DMSO-d6, ppm) δ 14.70 (s, 1H), 8.1 (bs, 2H), 7.64 (s, 1H), 7.60 – 7.53 (m, 2H), 7.25 (td, J = 8.5, 2.7 Hz, 1H), 7.08 (s, 1H), 5.77 (m, 1H), 4.55 (d, J = 5.1 Hz, 1H), 4.36 (d, J = 5.1 Hz, 1H), 3.02 (s, 3H), 1.73 (d, J = 6.2 Hz, 3H). HPLC purity: 85.0%; LCMS Calculated for C20H17FN6O2: 392.14; Observed: 393.2 [M+H]+.

[0340] Step-9. Synthesis of tert-butyl N-[(5-cyano-4-iodo-1H-pyrazol-3-yl)methyl]-N- methylcarbamate INT-17

[0341] To a stirred solution of tert-butyl N-[(5-cyano-1H-pyrazol-3-yl)methyl]-N- methylcarbamate (45.0 g, 190 mmol, 1.00 equiv) in MeCN (400 mL) was added NIS (42.9 g, 190 mmol, 1.00 equiv) in portions at room temperature. The resulting mixture was stirred for 16 hours at 80 . The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with ethyl acetate / petroleum ether (0 to 50% ethyl acetate) to afford tert-butyl N-[(5-cyano-4-iodo-1H-pyrazol-3-yl)methyl]-N-methylcarbamate INT-17 (40.0 g, 52%) as a yellow solid. INT-18. Synthesis of (R)-26-amino-55-fluoro-4,7-dimethyl-6-oxo-11-(piperidin-4-ylmethyl)- 11H-3-oxa-7-aza-2(3,5)-pyridina-1(4,3)-pyrazola-5(1,2)-benzenacyclooctaphane-15- carbonitrile.

[0342] Step-1. Synthesis of tert-butyl (R)-4-((26-amino-15-cyano-55-fluoro-4,7-dimethyl-6- oxo-11H-3-oxa-7-aza-2(3,5)-pyridina-1(4,3)-pyrazola-5(1,2)-benzenacyclooctaphane-11- yl)methyl) piperidine-1-carboxylate

[0343] To a stirred solution of (R)-26-amino-55-fluoro-4,7-dimethyl-6-oxo-11H-3-oxa-7-aza- 2(3,5)-pyridina-1(4,3)-pyrazola-5(1,2)-benzenacyclooctaphane-15-carbonitrile (300 mg, 764 µmol) and tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (197 mg, 916 µmol), PPh3(298 mg, 1.14 mmol) in THF (10 mL) was added DIAD (230 mg, 1.14 mmol) at 0 under nitrogen atmosphere. The resulting mixture was stirred for 3 hours at 0 under nitrogen atmosphere. The reaction was quenched with H2O (1 mL) at 0 . The resulting mixture was concentrated in vacuum. The residue was purified by prep-HPLC (NH3·H2O buffer) to afford tert-butyl (R)-4- ((26-amino-15-cyano-55-fluoro-4,7-dimethyl-6-oxo-11H-3-oxa-7-aza-2(3,5)-pyridina-1(4,3)- pyrazola-5(1,2)-benzenacyclooctaphane-11-yl)methyl)piperidine-1-carboxylate (200 mg, 339 µmol , 80% purity, 44% yield) as a yellow oil.1H NMR (300 MHz, DMSO-d6) δ 7.66 ̽ 7.56 (m, 3H), 7.50 ̽ 7.44 (m, 1H), 6.82 (d, J = 1.9 Hz, 1H), 5.62 (q, J = 6.8 Hz, 1H), 4.26 ̽ 4.22 (m, 2H), 3.99 ̽ 3.89 (m, 2H), 3.00 (s, 3H), 2.81 ̽ 2.61 (m, 3H), 2.18 ̽ 2.03 (m, 2H), 1.69 (d, J = 6.1 Hz, 3H), 1.61 ̽ 1.46 (m, 4H), 1.40 (s, 9H). HPLC purity: 80%; LCMS Calculated for: C31H36FN7O4:589.28; Observed: 590.3 [M+H]+. INT-19. Synthesis of 2-chloro-7-cyclopropylpyrazolo[1,5-a]pyrimidine-6-carboxylic acid

[0344] Step-1. Synthesis of ethyl (2Z)-2-[(Z)-cyclopropanecarbonyl]-3-ethoxyprop-2-enoate

[0345] Into a 250 mL 3-necked round-bottom was placed ethyl 3-cyclopropyl-3- oxopropanoate (5.00 g, 32.0 mmol) and (diethoxymethoxy)ethane (9.48 g, 64.0 mmol) in Ac2O(60 mL) at room temperature. The resulting mixture was stirred overnight at 130 °C under nitrogen atmosphere. The mixture was allowed to cool down to room temperature and concentrated in vacuum to afford ethyl (2Z)-2-[(Z)-cyclopropanecarbonyl]-3-ethoxyprop-2-enoate (5.77 g, 27.2 mmol, 82.4% purity, 84.5% yield) as a yellow oil, which was used for next step directly.

[0346] Step-2. Synthesis of ethyl 2-chloro-7-cyclopropylpyrazolo[1,5-a]pyrimidine-6- carboxylate

[0347] A solution of ethyl (2Z)-2-[(Z)-cyclopropanecarbonyl]-3-ethoxyprop-2-enoate (4.00 g, 18.8 mmol) and 3-chloro-1H-pyrazol-5-amine (2.20 g, 18.8 mmol) in EtOH (80 mL) was stirred for overnight at 80oC. The mixture was allowed to cool down to room temperature, the resulting mixture was filtered, the filter cake was washed with Et2O (3x 30 mL). The resulting mixture was concentrated in vacuum to afford ethyl 2-chloro-7-cyclopropylpyrazolo[1,5-a]pyrimidine-6- carboxylate (2.62 g, 9.89 mmol, 97.4% purity, 52.5% yield) as a white solid.1H NMR (300 MHz, DMSO-d6) δ 8.76 (s, 1H), 6.94 (s, 1H), 4.37 (q, J = 7.1 Hz, 2H), 2.98 – 2.84 (m, 1H), 1.73 – 164 (m, 2H), 1.36 (t, J = 7.1 Hz, 3H), 1.25 – 1.14 (m, 2H).

[0348] Step-3. Synthesis of 2-chloro-7-cyclopropylpyrazolo[1,5-a]pyrimidine-6-carboxylic acid (INT-19)

[0349] To a solution of ethyl 2-chloro-7-cyclopropylpyrazolo[1,5-a]pyrimidine-6-carboxylate (2.00 g, 7.52 mmol) in EtOH (20 mL) and H2O (20 mL) was added NaOH (889 mg, 22.5 mmol). The reaction mixture was stirred for 2 hours at room temperature. The resulting mixture was adjusted pH=3 with 2M HCl, the precipitated solids were collected by filtration and washed with water (3 x 20 mL). The resulting solid was dried under infrared light. This resulted in 2-chloro-7- cyclopropylpyrazolo[1,5-a]pyrimidine-6-carboxylic acid (INT-19). Yield: 1.8 g, 83%; HPLC purity: 98.6%; LCMS Calculated for C10H8ClN3O2: 237.03; Observed:238.1[M+H]+. INT-20. Synthesis of 1-(2-{4-[(3R)-3-aminopyrrolidin-1-yl]butoxy}-5-chloropyridin-3-yl)-3-{2- chloro-7-cyclopropylpyrazolo[1,5-a]pyrimidin-6-yl}urea.

[0350] Step-1. Synthesis of 4-[(5-chloro-3-nitropyridin-2-yl)oxy]butan-1-ol

[0351] A solution of tert-butyl N-(17-hydroxy-3,6,9,12,15-pentaoxaheptadecan-1- yl)carbamate (2.16 g, 5.67 mmol) and butane-1,4-diol (2.33 g, 25.9 mmol) in DMF (10.0 mL) was added t-BuOK (2.90 g, 25.9 mmol) at 0 . The mixture was stirred for 2 hours at 0 . The mixture was warmed to room temperature and concentrated in vacuum. The residue was purified by prep- HPLC (NH3·H2O buffer) to afford 4-[(5-chloro-3-nitropyridin-2-yl)oxy]butan-1-ol (2.56 g, 10.4 mmol, 86% purity, 40% yield) as a brown oil.

[0352] Step-2. Synthesis of 4-[(5-chloro-3-nitropyridin-2-yl)oxy]butanal

[0353] A solution of 4-[(5-chloro-3-nitropyridin-2-yl)oxy]butan-1-ol (2.00 g, 8.10 mmol) and IBX (2.94 g, 10.5 mmol) in MeCN (30 mL) was stirred for 2 hours at 80 . The mixture was allowed to cool down to room temperature and filtered. The filter cake was washed with MeCN (3x 30 mL) and the filtrate was concentrated under reduced pressure to afford crude 4-[(5-chloro-3-nitropyridin-2-yl)oxy]butanal (1.46 g, 6.00 mmol, 86% purity, 74% yield) as a yellow oil, which was used for next step directly.

[0354] Step-3. Synthesis of tert-butyl N-[(3R)-1-{4-[(5-chloro-3-nitropyridin-2- yl)oxy]butyl}pyrrolidin-3-yl]carbamate

[0355] To a solution of 4-[(5-chloro-3-nitropyridin-2-yl)oxy]butanal (1.80 g, 7.35 mmol) and tert-butyl N-[(3R)-pyrrolidin-3-yl]carbamate (1.36 g, 7.35 mmol) in MeOH (30 mL) was added NaBH3CN (1.49 g, 22.0 mmol). The reaction mixture was stirred for 2 hours at room temperature. The resulting mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic phase was washed with brine (30 mL), dried over Na2SO4, filtered and concentrated in vacuum. The residue was purified by silica gel chromatography (Petroleum / ethyl acetate = 1:1) to give tert-butyl N-[(3R)-1-{4-[(5-chloro-3-nitropyridin-2-yl)oxy]butyl}pyrrolidin- 3-yl]carbamate (1.20 g, 2.89 mmol, 92% purity, 40% yield) as a brown solid.

[0356] Step-4. Synthesis of tert-butyl N-[(3R)-1-{4-[(3-amino-5-chloropyridin-2- yl)oxy]butyl} pyrrolidin-3-yl]carbamate

[0357] Into a 250 mL round-bottom flask, was placed tert-butyl N-[(3R)-1-{4-[(5-chloro-3- nitropyridin-2-yl)oxy]butyl}pyrrolidin-3-yl]carbamate (1.00 g, 2.41 mmol) in MeOH (20 mL) was added Raney-Ni (282 mg, 4.82 mmol) at room temperature under nitrogen atmosphere and then the reaction mixture was degassed and purged with hydrogen for three times. The resulting reaction mixture was stirred for 2 hours under hydrogen atmosphere. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give tert-butyl N-[(3R)-1-{4-[(3-amino-5- chloropyridin-2-yl)oxy]butyl}pyrrolidin-3-yl]carbamate (725 mg, 1.88 mmol, 85% purity, 78% yield) as an off-white solid.

[0358] Step-5. Synthesis of tert-butyl N-[(3R)-1-[4-({5-chloro-3-[({2-chloro-7- cyclopropylpyrazolo[1,5-a]pyrimidin-6-yl}carbamoyl)amino]pyridin-2-yl}oxy)butyl]pyrrolidin- 3-yl]carbamate

[0359] Into a 40 mL 4-necked round-bottom was placed 2-chloro-7-cyclopropylpyrazolo[1,5- a]pyrimidine-6-carboxylic acid (306 mg, 1.29 mmol), DPPA (530 mg, 1.93 mmol) and TEA (390 mg, 3.87 mmol) in dioxane (10 mL) was stirred at room temperature for 30 min. tert-butyl N- [(3R)-1-{4-[(3-amino-5-chloropyridin-2-yl)oxy]butyl}pyrrolidin-3-yl]carbamate (500 mg, 1.29 mmol) was then added, and the reaction mixture was stirred for 4.0 hours at 100oC. The mixture was allowed to cool down to room temperature and the resulting mixture was filtered. The filtratewas concentrated in vacuum. The residue was purified by silica gel chromatography, eluted with petroleum ether : ethyl acetate =1:1 to afford tert-butyl N-[(3R)-1-[4-({5-chloro-3-[({2-chloro-7- cyclopropylpyrazolo[1,5-a]pyrimidin-6-yl}carbamoyl)amino]pyridin-2-yl}oxy)butyl]pyrrolidin- 3-yl]carbamate (113 mg, 183 µmol, 76% purity, 14% yield) as a brown solid.

[0360] Step-6. Synthesis of 1-(2-{4-[(3R)-3-aminopyrrolidin-1-yl]butoxy}-5-chloropyridin- 3-yl)-3-{2-chloro-7-cyclopropylpyrazolo[1,5-a]pyrimidin-6-yl}urea, INT-20

[0361] A solution of tert-butyl N-[(3R)-1-[4-({5-chloro-3-[({2-chloro-7- cyclopropylpyrazolo[1,5-a]pyrimidin-6-yl}carbamoyl)amino]pyridin-2-yl}oxy)butyl]pyrrolidin- 3-yl]carbamate (140 mg, 225 µmol) in TFA / DCM (1:1, 5 mL) was stirred for 1 hour at room temperature. The resulting mixture was concentrated in vacuum to afford 1-(2-{4-[(3R)-3- aminopyrrolidin-1-yl]butoxy}-5-chloropyridin-3-yl)-3-{2-chloro-7-cyclopropylpyrazolo[1,5- a]pyrimidin-6-yl}urea INT-20 as TFA salt (103 mg, 199 µmol, 88% purity, 94% yield) as a brown oil. INT-21 & INT-22. Synthesis of methyl (2E)-3-(4-{[(4S)-4-{[(3R)-1-acetylpyrrolidin-3- yl]carbamoyl}-4-{[(tert-butoxy)carbonyl]amino}butyl]amino}-3-nitrophenyl)prop-2-enoate, INT-22

[0362] Step-1. Synthesis of tert-butyl N-[(1S)-1-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}- 4-{[(benzyloxy)carbonyl]amino}butyl]carbamate

[0363] A mixture of (2S)-5-{[(benzyloxy)carbonyl]amino}-2-{[(tert- butoxy)carbonyl]amino}pentanoic acid (2.1 g, 5.73 mmol), 1-[(3R)-3-aminopyrrolidin-1- yl]ethan-1-one hydrochloride (1.03 g, 6.30 mmol) and DIEA (2.20 g, 17.1 mmol) in DMF (30.0 mL) was stirred at room temperature for 10 min. HATU (2.61 g, 6.87 mmol) was then added, and the reaction mixture was stirred for 1.0 hour at room temperature. The mixture diluted with water (200 mL) and extracted with ethyl acetate (200 mLx3). The combined organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuum. The residue was purified by silica gel column chromatography, eluted with petroleum ether: ethyl acetate (50% to 80% ethyl acetate) to give tert-butyl N-[(1S)-1-{[(3R)-1-acetylpyrrolidin-3- yl]carbamoyl}-4-{[(benzyloxy)carbonyl]amino} butyl]carbamate (2.00 g, 4.19 mmol, 95% purity, 73% yield) as a white solid.

[0364] Step-2. Synthesis of tert-butyl N-[(1S)-1-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}- 4-aminobutyl]carbamate, INT-21

[0365] To a solution of tert-butyl N-[(1S)-1-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}-4- {[(benzyloxy)carbonyl]amino}butyl]carbamate (2.0 g, 4.19 mmol) in MeOH (20 mL) was added Pd / C (445 mg, 4.19 mmol, 10%) under nitrogen atmosphere in a 100 mL round-bottom flask. The mixture was hydrogenated at room temperature for 3 hours under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure to give tert-butyl N-[(1S)-1-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}-4-aminobutyl]carbamate (1.40 g, 4.08 mmol, 90% purity, 95% yield) as colorless oil.

[0366] Step-3. Synthesis of methyl (2E)-3-(4-{[(4S)-4-{[(3R)-1-acetylpyrrolidin-3- yl]carbamoyl}-4-{[(tert-butoxy)carbonyl]amino}butyl]amino}-3-nitrophenyl)prop-2-enoate

[0367] Into a 100 mL 3-necked round-bottom was placed tert-butyl N-[(1S)-1-{[(3R)-1- acetylpyrrolidin-3-yl]carbamoyl}-4-aminobutyl]carbamate (900 mg, 2.62 mmol), methyl (2E)-3- (4-fluoro-3-nitrophenyl)prop-2-enoate (884 mg, 3.93 mmol), DIEA (1.01 g, 7.86 mmol) and 1,4- dioxane (10.0 mL) at room temperature. The resulting mixture was stirred overnight at 110 °C under nitrogen atmosphere (1atm). The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated in vacuum, the residue was purified by reverse flash chromatography with the following conditions: column, C18 silica gel; mobile phase, Aqueous phase of 0.05% ammonia in acetonitrile, 60% to 80% gradient in 20 min; detector, UV 254 nm. to afford methyl (2E)-3-(4-{[(4S)-4-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}-4-{[(tert-butoxy)carbonyl]amino} butyl]amino}-3-nitrophenyl)prop-2-enoate (1.00 g, 1.82 mmol, 90% purity, 69% yield) as a yellow solid.

[0368] Step-4. Synthesis of methyl methyl 3-(4-{[(4S)-4-{[(3R)-1-acetylpyrrolidin-3- yl]carbamoyl}-4-{[(tert-butoxy)carbonyl]amino}butyl]amino}-3-aminophenyl)propanoate

[0369] To a solution of methyl (2E)-3-(4-{[(4S)-4-{[(3R)-1-acetylpyrrolidin-3- yl]carbamoyl}-4-{[(tert-butoxy)carbonyl]amino}butyl]amino}-3-nitrophenyl)prop-2-enoate (600 mg, 1.09 mmol) in MeOH (12.0 mL) was added Pd / C (115 mg, 1.09 mmol, 10%) under nitrogen atmosphere in a 50 mL round-bottom flask. The mixture was hydrogenated at room temperature for 2 hours under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure to give methyl 3-(4-{[(4S)-4-{[(3R)-1-acetylpyrrolidin- 3-yl]carbamoyl}-4-{[(tert-butoxy)carbonyl]amino}butyl]amino}-3-aminophenyl)propanoate (450 mg, 865 µmol, 85% purity, 79% yield) a yellow oil.

[0370] Step-5. Synthesis of methyl 3-{1-[(4S)-4-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}- 4-{[(tert-butoxy)carbonyl]amino}butyl]-2-amino-1H-1,3-benzodiazol-5-yl}propanoate

[0371] Into a 50 mL round-bottom was placed methyl 3-(4-{[(4S)-4-{[(3R)-1- acetylpyrrolidin-3-yl]carbamoyl}-4-{[(tert-butoxy)carbonyl]amino}butyl]amino}-3- aminophenyl)propanoate (400 mg, 769 µmol), carbononitridic bromide (488 mg, 4.61 mmol) and EtOH (8.0 mL). The resulting mixture was stirred for 2 hours at room temperature. The resulting mixture was concentrated in vacuum, the residue was purified by reverse flash chromatography with the following conditions: column, C18 silica gel; mobile phase, Aqueous phase of 0.05% ammonia in acetonitrile, 40% to 70% gradient in 20 min; detector, UV 254 nm. to afford methyl 3-{1-[(4S)-4-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}-4-{[(tert- butoxy)carbonyl]amino}butyl]-2-amino-1H-1,3-benzodiazol-5-yl}propanoate (250 mg, 459 µmol, 80% purity, 59% yield) as white solid.

[0372] Step-6. Synthesis of methyl (2E)-3-(4-{[(4S)-4-{[(3R)-1-acetylpyrrolidin-3- yl]carbamoyl}-4-{[(tert-butoxy)carbonyl]amino}butyl]amino}-3-nitrophenyl)prop-2-enoate, (INT-22).

[0373] A solution of methyl 3-{1-[(4S)-4-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}-4- {[(tert-butoxy)carbonyl]amino}butyl]-2-amino-1H-1,3-benzodiazol-5-yl}propanoate (240 mg, 440 µmol) in THF: H2O (2:1, 4.8 mL) were added LiOH (18.4 mg, 1.76 mmol) at room temperature. The resulting mixture was stirred for 2 hours at room temperature. The mixture wasacidified to pH 2-3 with 1 M HCl. The precipitated solids were collected by filtration and washed with water (50 mL). This resulted in 3-{1-[(4S)-4-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}-4- {[(tert-butoxy)carbonyl]amino}butyl]-2-amino-1H-1,3-benzodiazol-5-yl}propanoic acid, (INT- 22). Yield: 210 mg, 90.1%; Appearance: White solid;1H NMR (300 MHz, DMSO-d6) δ 8.76 – 8.52 (m, 1H), 7.39 – 7.13 (m, 1H), 7.02 – 6.91 (m, 2H), 6.73 (d, J = 8.0 Hz, 1H), 6.20 (s, 2H), 4.27 – 4.07 (m, 1H), 3.52 – 3.08 (m, 9H), 2.86 – 2.70 (m, 2H), 2.17 (t, J = 7.7 Hz, 2H), 2.01 – 1.84 (m, 2H), 1.66 – 1.54 (m, 5H), 1.35 (s, 9H). HPLC purity: 92.4%; LCMS Calculated for C26H38N6O6: 530.29; Observed: 531.2[M+H]+. INT-23 & INT-24. Synthesis of (16R)-19-amino-4-(6-bromohexyl)-13-fluoro-8,16-dimethyl-9- oxo-17-oxa-4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2,5,10(15),11,13,18,20 - octaene-3-carbonitrile, INT-23 & (16R)-19-amino-5-(6-bromohexyl)-13-fluoro-8,16-dimethyl- 9-oxo-17-oxa-4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa- 1(22),2(6),3,10(15),11,13,18,20-octaene-3-carbonitrile, INT-24:

[0374] Step-1. Synthesis of (16R)-19-amino-4-(6-bromohexyl)-13-fluoro-8,16-dimethyl-9- oxo-17-oxa-4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2,5,10(15),11,13,18,20- octaene-3-carbonitrile

[0375] Into a 40 mL vial was placed (16R)-19-amino-13-fluoro-8,16-dimethyl-9-oxo-17-oxa- 4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2,5,10(15),11,13,18,20-octaene-3- carbonitrile (200 mg, 509 µmol), 1,6-dibromohexane (246 mg, 1.01 mmol), K2CO3(280 mg, 2.03 mmol) and MeCN (4.0 mL). The resulting reaction was allowed to heat to 60 ℃ for 3.0 hours. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated in vacuum, the residue was purified by reverse flash chromatography with the following conditions: column, C18 silica gel; mobile phase, Aqueous phase of 0.05% ammonia in acetonitrile, 70% to 100% gradient in 20 min; detector, UV 254 nm. to afford (16R)-19-amino-4- (6-bromohexyl)-13-fluoro-8,16-dimethyl-9-oxo-17-oxa-4,5,8,20- tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22), 2,5,10(15),11,13,18,20-octaene-3-carbonitrileINT-23 (150 mg, 270 µmol, 90% purity, 53% yield) as a white solid and (16R)-19-amino-5-(6- bromohexyl)-13-fluoro-8,16-dimethyl-9-oxo-17-oxa-4,5,8,20- tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2(6),3,10(15),11,13,18,20-octaene-3- carbonitrile INT-24 (70.0 mg, 126 µmol, 89% purity, 25% yield) as a white solid. INT-25. Synthesis of methyl 5-{[(1R)-1-(5-fluoro-2-hydroxyphenyl)ethyl]amino}pyrazolo [1,5- a]pyrimidine-3-carboxylate, INT-25

[0376] Step-1. Synthesis of methyl 5-{[(1R)-1-(5-fluoro-2- hydroxyphenyl)ethyl]amino}pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0377] A mixture of methyl 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate (2.20 g, 10.4 mmol), 2-[(1R)-1-aminoethyl]-4-fluorophenol hydrochloride (2.00 g, 10.4 mmol) and DIEA (4.02 g, 31.2 mmol) in MeCN (20 mL) at room temperature and the reaction mixture was stirred for 3 hours at 80 °C. The resulting mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by prep-HPLC (NH3.H2O buffer) to afford methyl 5-{[(1R)-1-(5-fluoro-2-hydroxyphenyl)ethyl]amino}pyrazolo[1,5- a]pyrimidine-3-carboxylate (2.30 g, 6.96 mmol, 85.0% purity, 53.2% yield) as a yellow solid.

[0378] Step-2. Synthesis of methyl 5-{[(1R)-1-(2-{[(2S)-1-{[(tert- butoxy)carbonyl]amino}propan-2-yl]oxy}-5-fluorophenyl)ethyl]amino}pyrazolo[1,5- a]pyrimidine-3-carboxylate

[0379] A mixture of methyl 5-{[(1R)-1-(5-fluoro-2- hydroxyphenyl)ethyl]amino}pyrazolo[1,5-a] pyrimidine-3-carboxylate (2.30 g, 6.96 mmol) , tert-butyl N-[(2R)-2-hydroxypropyl]carbamate (1.82 g, 10.4 mmol) and PPh3(2.72 g, 10.4 mmol) in THF (20 mL) and DCM (20 mL ) at 0 °C. Then to above mixture was added DTBAD (2.39 g, 10.4 mmol) at 0 °C under nitrogen atmosphere, and the reaction mixture was stirred for 2 hours at room temperature. The resulting mixture was quenched with H2O (5 mL) and concentrated under reduced pressure. The residue was purified by prep-HPLC (NH3.H2O buffer) to afford methyl 5-{[(1R)-1-(2-{[(2S)-1-{[(tert-butoxy)carbonyl]amino}propan-2-yl]oxy}-5-fluorophenyl)ethyl]amino}pyrazolo[1,5-a] pyrimidine-3-carboxylate (1.60 g, 3.28 mmol, 90.0% purity, 50.5% yield) as a yellow oil.

[0380] Step-3. Synthesis of methyl 5-{[(1R)-1-(2-{[(2S)-1-aminopropan-2-yl]oxy}-5- fluorophenyl) ethyl]amino}pyrazolo[1,5-a]pyrimidine-3-carboxylate, INT-25

[0381] A mixture of methyl 5-{[(1R)-1-(2-{[(2S)-1-{[(tert-butoxy)carbonyl]amino}propan- 2-yl]oxy}-5-fluorophenyl)ethyl]amino}pyrazolo[1,5-a]pyrimidine-3-carboxylate (1.60 g, 3.28 mmol) in HCl / dioxane (20 mL, 4M) at room temperature, the reaction mixture was stirred for 1 hour at room temperature. The resulting mixture was concentrated under reduced pressure to afford methyl 5-{[(1R)-1-(2-{[(2S)-1-aminopropan-2-yl]oxy}-5-fluorophenyl)ethyl]amino} pyrazolo[1,5-a]pyrimidine-3-carboxylate as HCl salt (1.40 g, 3.61 mmol, crude) as a yellow solid, which was used for next step directly. INT-26. Synthesis of 2-[(16R)-19-amino-3-cyano-13-fluoro-8,16-dimethyl-9-oxo-17-oxa- 4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2,5,10(15),11,13,18,20-octaen-4- yl]acetic acid.

[0382] Step-1. Synthesis of tert-butyl 2-[(16R)-19-amino-3-cyano-13-fluoro-8,16-dimethyl- 9-oxo-17-oxa-4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2,5,10(15),11,13,18,20- octaen-4-yl]acetate

[0383] A mixture of (16R)-19-amino-13-fluoro-8,16-dimethyl-9-oxo-17-oxa-4,5,8,20- tetraaza tetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2,5,10(15),11,13,18,20-octaene-3-carbonitrile (400 mg, 1.01 mmol), K2CO3(418 mg, 3.03 mmol), tert-butyl 2-bromoacetate (294 mg, 1.51 mmol) and DMF (4 mL) at room temperature. The resulting mixture was stirred for 3 hours at room temperature. The residue was purified by reverse flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water, 30% to 70% gradient in 20 min; detector, UV 254 nm. This resulted in tert-butyl 2-[(16R)-19-amino-3-cyano-13-fluoro-8,16- dimethyl-9-oxo-17-oxa-4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2,5,10(15), 11,13,18,20-octaen-4-yl]acetate (200 mg, 394 µmol, 85% purity, 39% yield) as a yellow solid and tert-butyl (R)-2-(26-amino-13-cyano-55-fluoro-4,7-dimethyl-6-oxo-11H-3-oxa-7-aza-2(3,5)-pyridina-1(4,5)-pyrazola-5(1,2)-benzenacyclooctaphane-11-yl)acetate (90 mg, 184 µmol, 85% purity, 18% yield) as a yellow solid.

[0384] Step-2. Synthesis of 2-[(16R)-19-amino-3-cyano-13-fluoro-8,16-dimethyl-9-oxo-17- oxa-4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2,5,10(15),11,13,18,20-octaen-4- yl]acetic acid, INT-26

[0385] A solution of tert-butyl 2-[(16R)-19-amino-3-cyano-13-fluoro-8,16-dimethyl-9-oxo- 17-oxa-4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵]docosa-1(22),2,5,10(15),11,13,18,20-octaen- 4-yl]acetate (200 mg, 394 µmol) in TFA:DCM (1:1, 2 mL) was stirred for 12.0 hours at room temperature. The resulting mixture was concentrated in vacuum to afford 2-[(16R)-19-amino-3- cyano-13-fluoro-8,16-dimethyl-9-oxo-17-oxa-4,5,8,20-tetraazatetracyclo[16.3.1.0²,⁶.0¹⁰,¹⁵] docosa-1(22),2,5,10(15),11,13,18,20-octaen-4-yl]acetic acid as TFA salt (170 mg, 377 µmol, crude) as a yellow oil, which was used for next step directly. INT-27. Synthesis of (R)-6-(7-((tert-butoxycarbonyl)amino)-5-azaspiro[2.4]heptan-5- yl)hexanoic acid.

[0386] Step-1. Synthesis of methyl (R)-6-(7-((tert-butoxycarbonyl)amino)-5- azaspiro[2.4]heptan-5-yl)hexanoate

[0387] To a stirred solution of tert-butyl N-[(7R)-5-azaspiro[2.4]heptan-7-yl]carbamate (500 mg, 2.35 mmol) and methyl 6-bromohexanoate (491 mg, 2.35 mmol), K2CO3(972 mg, 7.05 mmol) in MeCN (5 mL) at room temperature. The resulting mixture was stirred for 2 hours at 80 °C. The mixture was allowed to cool down to room temperature. The residue was purified by reverse phase flash with the following conditions (column, C18 silica gel; mobile phase, H2O / 0.1% NH4OH in MeCN, 10% to 60% gradient in 12 min; detector, UV 254 nm.) to afford methyl (R)-6-(7-((tert- butoxycarbonyl)amino)-5-azaspiro[2.4]heptan-5-yl)hexanoate (780 mg, 2.29 mmol, 91.2% purity, 97.5% yield) as a yellow solid.

[0388] Step-2. Synthesis of (R)-6-(7-((tert-butoxycarbonyl)amino)-5-azaspiro[2.4]heptan-5- yl)hexanoic acid, INT-27

[0389] A solution of methyl (R)-6-(7-((tert-butoxycarbonyl)amino)-5-azaspiro[2.4]heptan-5- yl)hexanoate (780 mg, 2.29 mmol) in THF: H2O (2:1, 16 mL) were added NaOH (274 mg, 6.86 mmol) at room temperature. The resulting mixture was stirred for 1.0 hour at room temperature. The mixture was acidified to pH 2-3 with 1 M HCl. The precipitated solids were collected by filtration and washed with water (50 mL). This resulted in (R)-6-(7-((tert-butoxycarbonyl)amino)- 5-azaspiro[2.4]heptan-5-yl)hexanoic acid, INT-27. Yield: 720 mg, crude; Appearance: White solid; HPLC purity: 88.1%; LCMS Calculated for C17H30N2O4: 326.22; Observed: 327.1[M+H]+. INT-28. Synthesis of 4-[(4R)-4-{[( tert-butoxy)carbonyl]amino}-3,3-dimethylpyrrolidin-1- yl]butanoic acid.

[0390] Step-1. Synthesis of methyl 4-[(4R)-4-{[( tert-butoxy)carbonyl]amino}-3,3- dimethylpyrrolidin-1-yl]butanoate

[0391] Into a 50 mL round-bottom was placed tert-butyl N-[(3R)-4,4-dimethylpyrrolidin-3- yl]carbamate hydrochloride (200 mg, 797 µmol), methyl 4-bromobutanoate (25 mg, 1.19 mmol), K2CO3(329 mg, 2.39 mmol) and MeCN (4 mL). The resulting mixture was stirred for 5.0 hours at 80°C under nitrogen atmosphere. The mixture was allowed to cool down to room temperature, diluted with water (50 mL) and extracted with ethyl acetate (50 mLx3). The combined organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuum to afford crude methyl 4-[(4R)-4-{[(tert-butoxy)carbonyl]amino}-3,3- dimethyl pyrrolidin-1-yl]butanoate (210 mg, 667 µmol, 70% purity, 84% yield) as a yellow oil. Step-2. Synthesis of 4-[(4R)-4-{[(tert-butoxy)carbonyl]amino}-3,3-dimethylpyrrolidin-1- yl]butanoic acid, INT-28

[0392] A mixture of methyl 4-[(4R)-4-{[(tert-butoxy)carbonyl]amino}-3,3- dimethylpyrrolidin-1-yl] butanoate (110 mg, 349 µmol) and LiOH·H2O (43.6 mg, 1.04 mmol) in THF / H2O (2:1, 3 mL) was stirred at room temperature for 3.0 hours. The resulting mixture was concentrated under reduced pressure, the crude product was purified by prep-HPLC (NH3.H2Obuffer) to give 4-[(4R)-4-{[( tert-butoxy)carbonyl]amino}-3,3-dimethylpyrrolidin-1-yl]butanoic acid (100 mg, 332 µmol, 75% purity, 85% yield) as a white solid. Example A1. Synthesis of (S)-N-((R)-1-acetylpyrrolidin-3-yl)-2-amino-5- guanidinopentanamide (A-1):

[0393] Step-1. Synthesis of tert-butyl (R)-(1-acetylpyrrolidin-3-yl)carbamate

[0394] To a stirred solution of tert-butyl N-[(3R)-pyrrolidin-3-yl]carbamate (5.00 g, 26.8 mmol) and pyridine (6.34 g, 80.3 mmol) in DCM (50 mL) was added AcCl (2.50 g, 32.1 mmol) at 0 °C. After addition, the reaction mixture was stirred at room temperature for 3 hours. The resulting mixture was diluted with water (50 mL) and extracted with DCM (50 mLx2). The combined organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuum. The residue was purified by silica gel chromatography, diluted with petroleum ether: ethyl acetate =1:2 to afford tert-butyl N-[(3R)-1-acetylpyrrolidin-3-yl]carbamate (4.59 g, 20.1 mmol, 90% purity, 75% yield) as a white solid.

[0395] Step-2. Synthesis of (R)-1-(3-aminopyrrolidin-1-yl)ethan-1-one

[0396] A solution of tert-butyl N-[(3R)-1-acetylpyrrolidin-3-yl]carbamate (5.10 g, 22.3 mmol) in TFA (50 mL) was stirred for 2.0 hours at room temperature. The resulting mixture was concentrated in vacuum to afford crude product 1-[(3R)-3-aminopyrrolidin-1-yl]ethan-1-one as TFA salt (2.97 g, 23.1 mmol, crude) as a brown oil.

[0397] Step-3. Synthesis of tert-butyl N-[(1S)-1-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}- 4-{[(Z)-{[(tert-butoxy)carbonyl]amino}({[(tertbutoxy)carbonyl]imino})methyl]amin o}butyl]carbamate

[0398] To a stirred solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(Z)-{[(tert- butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]amino}pentanoic acid (500 mg, 1.05 mmol), 1-[(3R)-3-aminopyrrolidin-1-yl]ethan-1-one (134 mg, 1.05 mmol) and DIEA (134mg, 1.05 mmol) in DCM (10 mL) was added HATU (399 mg, 1.05 mmol) at -40 degrees C. After 2.0 hours, the reaction was warmed to room temperature and concentrated in vacuum. The residue was purified by prep-HPLC (NH3.H2O buffer) to afford tert-butyl N-[(1S)-1-{[(3R)-1- acetylpyrrolidin-3-yl]carbamoyl}-4-{[(Z)-{[(tert-butoxy)carbonyl]amino}({[(tert- butoxy)carbonyl]imin o})methyl]amino}butyl]carbamate (424 mg, 725 µmol, 90% purity, 69% yield) as a light yellow solid.

[0399] Step-4. Synthesis of (S)-N-((R)-1-acetylpyrrolidin-3-yl)-2-amino-5- guanidinopentanamide, (A-1):

[0400] A solution of tert-butyl N-[(1S)-1-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}-4-{[(Z)- {[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]amino}butyl] carbamate (450 mg, 769 µmol) in dioxane (5 mL) was added dropwise HCl / dioxane (2 M in dioxane, 3.8 mL, 7.68 mmol) and stirred at room temperature for 3.0 h. The resulting mixture was concentrated in vacuum to afford (2S)-N-[(3R)-1-acetylpyrrolidin-3-yl]-2-amino-5-carbamimidamido pentanamide as HCl salt (A-1). Yield: 99.9 mg, 45.8%; Appearance: a brown semi-solid;1H NMR (300 MHz, DMSO-d6) δ 4.34 – 4.17 (m, 1H), 3.70 – 3.22 (m, 4H), 3.20 – 3.12 (m, 2H), 2.20 – 1.96 (m, 1H), 1.97 – 1.91 (m, 3H), 1.80 – 1.71 (m, 4H), 1.57 – 1.43 (m, 2H). HPLC purity: 97.0%; LCMS Calculated for C12H24N6O2: 284.20; Observed: 285.20[M+H]+.

[0401] The following examples were prepared using standard chemical manipulations and procedures similar to those used for the preparation of the previous examples. Analytical data is given in the table below.Example A2. Scheme 3. Synthesis of (S)-2-amino-5-guanidino-N-((S)-1-(methylamino)-1-oxo- 3-(tetrahydro-2H-pyran-4-yl)propan-2-yl)pentanamid (A-23):

[0402] Step-1. Synthesis of tert-butyl (S)-(1-(methylamino)-1-oxo-3-(tetrahydro-2H-pyran-4- yl)propan-2-yl)carbamate

[0403] To a solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-(oxan-4-yl)propanoic acid (200 mg, 0.731 mmol), MeNH2yHCl (98.5 mg, 1.46 mmol), HATU (333 mg, 0.877 mmol) and DIEA (376 mg, 2.92 mmol) in DMF (3 mL) was stirring for 3 h at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate (1:1) to afford tert-butyl N-[(1S)-1- (methylcarbamoyl)-2-(oxan-4-yl)ethyl]carbamate (200 mg, 0.698 mmol, 80% purity, 95% yield) as a white solid.

[0404] Step-2. Synthesis of (S)-2-amino-N-methyl-3-(tetrahydro-2H-pyran-4- yl)propanamide hydrochloride

[0405] To a solution of tert-butyl N-[(1S)-1-(methylcarbamoyl)-2-(oxan-4-yl)ethyl]carbamate (100 mg, 0.349 mmol) in HCl / dioxane (4M, 1.2 mL) was stirring for 1 hour at room temperature. The resulting mixture was concentrated under reduced pressure, the crude product (2S)-2-amino- N-methyl-3-(oxan-4-yl)propenamide as HCl salt (80.0 mg, 359 µmol, 80% purity, 98% yield) as a white solid, which was used in the next step directly without further purification.

[0406] Step-3. Synthesis of tert-butyl N-[(Z)-{[(tert-butoxy)carbonyl]amino}({[(4S)-4- {[(tert-butoxy)carbonyl]amino}-4-{[(1S)-1-(methylcarbamoyl)-2-(oxan-4- yl)ethyl]carbamoyl}butyl]amino})methylidene]carbamate

[0407] To a solution of (2S)-2-amino-N-methyl-3-(oxan-4-yl)propanamide hydrochloride (70 mg, 0.314 mmol), (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(Z)-{[(tert- butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]amino}pentanoic acid, INT-1(123 mg, 0.261 mmol), HATU (119 mg, 0.314 mmol) and DIEA (167 mg, 1.30 mmol) in DMF (1 mL) was stirring for 2 hours at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (NH3·H2O buffer) to afford tert-butyl N-[(Z)-{[(tert-butoxy)carbonyl]amino}({[(4S)-4-{[(tert-butoxy)carbonyl]amino}-4-{[(1S)-1- (methylcarbamoyl)-2-(oxan-4-yl)ethyl]carbamoyl}butyl]amino})methylidene]carbamate (60.0 mg, 0.933 mmol, 92% purity, 29% yield) as a white solid.

[0408] Step-4. Synthesis of (S)-2-amino-5-guanidino-N-((S)-1-(methylamino)-1-oxo-3- (tetrahydro-2H-pyran-4-yl)propan-2-yl)pentanamide (A-23)

[0409] To a solution of tert-butyl N-[(Z)-{[(tert-butoxy)carbonyl]amino}({[(4S)-4-{[(tert- butoxy)carbonyl]amino}-4-{[(1S)-1-(methylcarbamoyl)-2-(oxan-4-yl)ethyl]carbamoy l}butyl]amino})methylidene]carbamate (50 mg, 0.777 mmol) in HCI / dioxane (4M, 0.5 mL) was stirring for 1 hour at room temperature. The resulting mixture was concentrated under reduced pressure, the residue was purified by prep-HPLC (0.02% HCl Water phase) to afford (2S)-2- amino-5-carbamimidamido-N-[(1S)-1-(methylcarbamoyl)-2-(oxan-4-yl)ethyl]pentanamide as HCl salt (A-23). Yield: 15.0 mg, 56%; Appearance: White solid.1H NMR (300 MHz, DMSO- d6) δ 8.72 (d, J = 7.8 Hz, 1H), 8.41 – 8.25 (m, 3H), 8.11 – 8.02 (m, 1H), 7.87 – 7.78 (m, 1H), 4.37 – 4.24 (m, 1H), 3.90 – 3.80 (m, 3H), 3.32 – 3.20 (m, 2H), 3.19 – 3.09 (m, 2H), 2.59 (d, J = 4.5 Hz, 3H), 1.83 – 1.70 (m, 2H), 1.65 – 1.46 (m, 7H), 1.21– 1.05 (m, 2H). HPLC purity: 98%; LCMS Calculated for C15H30N6O3: 342.24; Observed: 343.3 [M+H]+. Example A3. Synthesis of (2S,3S)-2-((S)-2-amino-3-(6-aminopyridin-3-yl)propanamido)-N,3- dimethylpentanamide, (A-24):

[0410] Step-1. Synthesis of methyl (S)-3-(6-aminopyridin-3-yl)-2-((tert- butoxycarbonyl)amino) propanoate

[0411] BrCH2CH2Br (259 mg, 1.38 mmol) was added to a stirred suspension of Zn (1.79 g, 27.6 mmol) in dry DMF (30 mL) , and the mixture was stirred at 50 °C for 30 min. The reaction mixture was allowed to cool to room temperature. TMS-Cl (29.9 mg, 276 µmol) was added to the mixture, and the mixture was stirred for a further 30 min vigorously. methyl (2R)-2-{[(tert- butoxy)carbonyl]amino}-3-iodopropanoate (1.70 g, 5.19 mmol) in DMF was added to the reaction mixture. The reaction mixture was continued to stir at room temperature for 2 h. The reaction mixture was then standing for another 30 min, the supernatant liquid was transferred to the mixture of 5-bromopyridin-2-amine (600 mg, 3.46 mmol), Pd2(dba)3 (79.2 mg, 86.5 µmol) S-phos (71.0 mg, 173 µmol) via syringe. The reaction mixture was stirred at 50 °C for 16 h. After pouring into water, the mixture was extracted with ethyl acetate (2×50 mL). The combined organic layers were washed with brine (2×50 mL), dried over sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (MeOH / DCM = 0-10%) to give methyl (2S)-3- (6-aminopyridin-3-yl)-2-{[(tert-butoxy)carbonyl]amino}propanoate (650 mg, 2.20 mmol) as a white solid.

[0412] Step-2. Synthesis of (S)-3-(6-aminopyridin-3-yl)-2-((tert- butoxycarbonyl)amino)propanoic acid

[0413] To a solution of methyl (2S)-3-(6-aminopyridin-3-yl)-2-{[(tert-butoxy)carbonyl] amino}propanoate (640 mg, 2.16 mmol) in THF (5 mL) and H2O (1 mL) was added LiOH (258 mg, 10.8 mmol) at rt. The reaction mixture was stirred for 2 h at rt. The reaction mixture wasadjusted pH to 5-6 with 1 N HCl (aq) and concentrated under reduced pressure to give a residue which was purified by reverse combi flash (Biotage, 130 g Agela C18, MeCN / H2O(0.5% FA) to give (2S)-3-(6-aminopyridin-3-yl)-2-{[(tert-butoxy)carbonyl]amino}propanoic acid (300 mg, 1.06 mmol) as a white solid.

[0414] Step-3. Synthesis of tert-butyl ((S)-3-(6-aminopyridin-3-yl)-1-(((2S,3S)-3-methyl-1- (methylamino)-1-oxopentan-2-yl)amino)-1-oxopropan-2-yl)carbamate

[0415] To a solution of (2S)-3-(6-aminopyridin-3-yl)-2-{[(tert- butoxy)carbonyl]amino}propanoic acid (270 mg, 959 µmol), (2S,3S)-2-amino-N,3- dimethylpentanamide (115 mg, 797 µmol) and DIPEA (514 mg, 3.98 mmol) in DMF (5 mL) was added DEPBT (261 mg, 876 µmol) at rt. The reaction mixture was stirred for 16 h at rt. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by reverse combi flash (Biotage, 10 g Agela C18, MeCN / H2O(0.1% NH4HCO3) to give tert-butyl N-[(1S)-2-(6-aminopyridin-3-yl)-1-{[(1S,2S)-2-methyl-1- (methylcarbamoyl)butyl]carbamoyl}ethyl]carbamate (40.0 mg, 98.1 µmol) as a white solid.

[0416] Step-4. Synthesis of (2S,3S)-2-((S)-2-amino-3-(6-aminopyridin-3-yl)propanamido)- N,3-dimethyl pentanamide

[0417] A solution of tert-butyl N-[(1S)-2-(6-aminopyridin-3-yl)-1-{[(1S,2S)-2-methyl-1- (methyl carbamoyl)butyl]carbamoyl}ethyl]carbamate (120 mg, 294 µmol) in 4 M HCl (1,4- dioxane solution) (5 mL) was stirred for 1 h at rt. The reaction mixture was concentrated under reduced pressure to give a residue, which was further purified by Prep-HPLC (ACN / water / 0.1% NH4HCO3) to give (2S, 3S)-2-[(2S)-2-amino-3-(6-aminopyridin-3-yl)propanamido]-N,3- dimethylpentanamide (A-24). Yield: 12.2 mg, 13%; Appearance: White solid;1H NMR (400 MHz, DMSO) δ 7.96 – 7.79 (m, 2H), 7.72 (d, J = 2.0 Hz, 1H), 7.21 (dd, J = 8.4, 2.4 Hz, 1H), 6.34 (d, J = 8.4 Hz, 1H), 5.66 (s, 2H), 4.09 (t, J = 8.0 Hz, 1H), 2.76 – 2.65 (m, 1H), 2.57 (d, J = 4.8 Hz, 3H), 2.47 – 2.31 (m, 2H), 1.77 (s, 2H), 1.66 – 1.57 (m, 1H), 1.43 – 1.28 (m, 1H), 1.04 – 0.89 (m, 1H), 0.85 – 0.70 (m, 6H); HPLC purity: 93%; LCMS Calculated for C15H25N5O2: 307.20; Observed: 308.4 [M+H]+. Example A4. Synthesis of (2S)-N-[(3R)-1-acetylpyrrolidin-3-yl]-2-amino-5-(2-aminopyridin-3- yl) pentanamide (A-25):

[0418] Step-1. Synthesis of tert-butyl N-[(3R)-1-acetylpyrrolidin-3-yl]carbamate

[0419] To a solution of tert-butyl N-[(3R)-pyrrolidin-3-yl]carbamate (200 mg, 1.07 mmol) in DCM (5 mL) was added TEA (324 mg, 3.21 mmol), and acetyl chloride (125 mg, 1.60 mmol). The mixture solution was stirred at RT for 2 h. The reaction mixture was concentrated and purified by flash silica gel chromatography (MeOH-DCM = 0%-5%) to give tert-butyl N-[(3R)-1- acetylpyrrolidin-3-yl]carbamate (200 mg, 0.88 mmol, 100% purity, 81.9% yield) as colorless oil.

[0420] Step-2. Synthesis of 1-[(3R)-3-aminopyrrolidin-1-yl] ethan-1-one

[0421] To a solution of tert-butyl N-[(3R)-1-acetylpyrrolidin-3-yl]carbamate (200 mg, 0.88 mmol) in DCM (3 mL) was added TFA (1mL). The mixture solution was stirred at RT for 1 h.The reaction mixture was concentrated to give 1-[(3R)-3-aminopyrrolidin-1-yl]ethan-1-one (112 mg, 0.88 mmol, 100% purity, 100% yield) as colorless oil.

[0422] Step-3. Synthesis of tert-butyl N-[(1S)-1-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}- 4-(2-aminopyridin-3-yl)butyl]carbamate

[0423] To a solution of 1-[(3R)-3-aminopyrrolidin-1-yl]ethan-1-one (50 mg, 0.39 mmol) in DMF (5 mL) was added DIPEA (152 mg, 1.17 mmol) , (2S)-5-(2-aminopyridin-3-yl)-2-{[(tert- butoxy)carbonyl]amino}pentanoic acid, INT-2 (120 mg, 0.39 mmol) and DEPBT (139 mg, 0.47 mmol). The mixture solution was stirred at RT for 16 h. The reaction mixture was poured into water (20 mL), extracted with EA(20 mLx3). The combined organic layer was washed with anhydrous sodium sulfate and filtrated. The filtrate was concentrated and purification by silica gel chromatography (MeOH-DCM = 0%-20%) to give tert-butyl N-[(1S)-1-{[(3R)-1- acetylpyrrolidin-3-yl]carbamoyl}-4-(2-aminopyridin-3-yl)butyl]carbamate (60.0 mg, 0.14 mmol, 100% purity, 36.8% yield) as a yellow solid.

[0424] Step-4. Synthesis of (2S)-N-[(3R)-1-acetylpyrrolidin-3-yl]-2-amino-5-(2- aminopyridin-3-yl) pentanamide, A-25.

[0425] To a solution of tert-butyl N-[(1S)-1-{[(3R)-1-acetylpyrrolidin-3-yl]carbamoyl}-4-(2- aminopyridin-3-yl)butyl]carbamate (40 mg, 0.10 mmol) in DCM (2 mL) was added TFA (1 mL). The mixture solution was stirred at RT for 2 h.The mixture was concentrated in vacuum to give crude compound. The mixture was diluted with MeOH, and adjusted to pH 9 with NH4OH. The mixture was further purified by pre-HPLC (1 / 1000 NH4HCO3, acid method) to give (2S)-N-[(3R)- 1-acetylpyrrolidin-3-yl]-2-amino-5-(2-aminopyridin-3-yl)pentanamide (A-25). Yield: 15.4 mg, 50.6%; Appearance: White solid; Yield: 16.0mg, 50.6%; Appearance: White solid;1H NMR (400 MHz, DMSO-d6) δ 8.17 - 7.97 (m, 1H), 7.77 (dd, J = 4.8, 1.6 Hz, 1H), 7.17 (d, J = 7.2 Hz, 1H), 6.46 (dd, J = 7.2, 5.0 Hz, 1H), 5.65 (d, 2H), 4.30 -4.15 (m, 1H), 3.96 -3.61 (m, 1H), 3.49-3.42 (m, 2H), 3.24 - 3.11 (m, 2H), 2.37 (t, J = 6.8 Hz, 2H), 2.11 -1.94 (m, 1H), 1.91 (d, J = 8.4 Hz, 3H), 1.85-1.69 (m, 1H), 1.63 -1.36 (m, 4H); HPLC purity: 96.52%; LCMS Calculated for C16H25N5O2: 319.41 Observed: 320.2 [M+H]+.

[0426] The following example was prepared using standard chemical manipulations and procedures similar to those used for the preparation of the previous examples. Analytical data is given in the table below.Example A5. Synthesis of (2S,3S)-2-((S)-2-amino-3-(1H-pyrrolo[2,3-b]pyridin-3- yl)propanamido)-N,3-dimethylpentanamide, (A-27) and (2S,3S)-2-((R)-2-amino-3-(1H- pyrrolo[2,3-b]pyridin-3-yl)propanamido)-N,3-dimethylpentanamide (A-28):

[0427] Step-1. Synthesis of 3-bromo-1-(4-methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine

[0428] To a solution of 3-bromo-1H-pyrrolo[2,3-b]pyridine (1.00 g, 5.07 mmol) in Dichloromethane (5 mL) were added Tosyl chloride (966 mg, 5.07 mmol), 4- Dimethylaminopyridine (619 mg, 5.07 mmol) and Triethylamine (513 mg, 5.07 mmol). The resulting mixture was stirred at rt for 2 h. The reaction mixture was poured into water (10 mL), extracted with DCM (20 mL x 3). The combined organic layer was washed with brine (10 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by flash silica gel chromatography (SiO2, Pet. Ether / EtOAc = 10 / 3) to give 3-bromo-1-(4- methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine (900 mg, 2.56 mmol, 100% purity, 56% yield) as a white solid.

[0429] Step-2. Synthesis of methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-[1-(4- methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]propanoate

[0430] 1,2-Dibromoethane (255 mg, 1.36 mmol) was added to a stirred suspension of Zinc powder (1.77 g, 27.2 mmol) in DMF (5 mL) , and the mixture was stirred at 50 °C for 30 min. The reaction mixture was allowed to cool to rt. Chlorotrimethylsilane (29 mg, 272 µmol) was added to the mixture, and the mixture was stirred for a further 30 min vigorously. methyl (2R)-2-{[(tert- butoxy)carbonyl]amino}-3-iodopropanoate (1.11 g, 3.40 mmol) in DMF (5 mL) was added to the reaction mixture, which was then stirred at rt for 2 h. The reaction mixture was then standing foranother 30 min, the supernatant liquid was transferred to the 3-bromo-1-(4- methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridine (600 mg, 1.70 mmol), Tris(dibenzylideneacetone)dipalladium (92.4 mg, 101 µmol) and X-Phos (96.7 mg, 203 µmol) via syringe. The reaction mixture was stirred at 50 °C for 16 h. The reaction mixture was poured into water (20 mL), extracted with EtOAc (60 mL x 3). The combined organic layer was washed with brine (20 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by flash silica gel chromatography (SiO2, Pet. Ether / EtOAc = 10 / 7) to give methyl (2S)- 2-{[(tert-butoxy)carbonyl]amino}-3-[1-(4-methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-3- yl]propanoate (450 mg, 950 µmol, 87% purity, 56% yield) as a white solid.

[0431] Step-3. Synthesis of (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-[1-(4- methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]propanoic acid

[0432] To a solution of methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-[1-(4- methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]propanoate (450 mg, 950 µmol) in Tetrahydrofuran (3 mL) were added Lithium hydroxide (22.7 mg, 950 µmol) and Water (1 mL). The resulting mixture was stirred at rt for 1 h. The reaction mixture was concentrated to give (2S)- 2-{[(tert-butoxy)carbonyl]amino}-3-[1-(4-methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-3- yl]propanoic acid (436 mg, 948 µmol, 100% purity, 100% yield) as a white solid.

[0433] Step-4. Synthesis of tert-butyl N-[(1S)-1-{[(1S,2S)-2-methyl-1- (methylcarbamoyl)butyl] carbamoyl}-2-[1-(4-methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin- 3-yl]ethyl]carbamate

[0434] To a solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-[1-(4- methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]propanoic acid (450 mg, 979 µmol) in N,N-Dimethylformamid (10 mL) were added DEPBT (583 mg, 1.95 mmol), N,N- Diisopropylethylamine (126 mg, 979 µmol) and (2S,3S)-2-amino-N,3-dimethylpentanamide (210 mg, 1.46 mmol). The resulting mixture was stirred at rt for 16 h. The reaction mixture was poured into water (50 mL), extracted with EtOAc (100 mL x 3). The combined organic layer was washed with brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and purified by flash silica gel chromatography (SiO2, Pet. Ether / EtOAc = 5 / 4) to give tert-butyl N-[(1S)-1-{[(1S,2S)-2-methyl-1-(methylcarbamoyl)butyl]carbamoyl}-2-[1-(4- methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]ethyl]carbamate (380 mg, 648 µmol, 89% purity, 66% yield) as a yellow solid.

[0435] Step-5. Synthesis of tert-butyl N-[(1S)-1-{[(1S,2S)-2-methyl-1- (methylcarbamoyl)butyl] carbamoyl}-2-{1H-pyrrolo[2,3-b]pyridin-3-yl}ethyl]carbamate

[0436] To a solution of tert-butyl N-[(1S)-1-{[(1S,2S)-2-methyl-1- (methylcarbamoyl)butyl]carbamoyl}-2-[1-(4-methylbenzenesulfonyl)-1H-pyrrolo[2,3-b]pyridin- 3-yl]ethyl]carbamate (450 mg, 768 µmol) in Methanol (5 mL ) was added Potassium carbonate (530 mg, 3.84 mmol). The resulting mixture was stirred at 65 °C for 16 h. The reaction mixture was filtered and the filtrate concentrated to give tert-butyl N-[(1S)-1-{[(1S,2S)-2-methyl-1- (methylcarbamoyl)butyl]carbamoyl}-2-{1H-pyrrolo[2,3-b]pyridin-3-yl}ethyl]carbamate (330 mg, 764 µmol, 100% purity, 99% yield) as a white solid.

[0437] Step-6. Synthesis of (2S,3S)-2-[(2S)-2-amino-3-{1H-pyrrolo[2,3-b]pyridin-3- yl}propanamido]-N,3-dimethylpentanamide (16.5 mg, 49.7 µmol) and (2S,3S)-2-[(2R)-2-amino- 3-{1H-pyrrolo[2,3-b]pyridin-3-yl}propanamido]-N,3-dimethylpentanamide

[0438] To a solution of tert-butyl N-[(1S)-1-{[(1S,2S)-2-methyl-1- (methylcarbamoyl)butyl]carbamoyl}-2-{1H-pyrrolo[2,3-b]pyridin-3-yl}ethyl]carbamate (100 mg, 231 µmol) was added HCl-dioxane (729 mg, 20.0 mmol) . The resulting mixture was stirred at rt for 1 h. Adjusted the pH=7-8 with NH4OH and filtered. The reaction solution was concentrated and purified with prep-HPLC(ACN / H2O / 0.5% NH4HCO3) to give crude product, which was sent for SFC (Mobile phase: CO2 / EtOH[1%NH3(7M in MeOH)]=60 / 40. Flow rate: 3 ml / min. Back pressure: 2000 psi. Column: Daicel AD-3 (4.6*100mm 3um)) to give (2S,3S)-2-[(2S)-2-amino-3- {1H-pyrrolo[2,3-b]pyridin-3-yl}propanamido]-N,3-dimethylpentanamide (A-27). Yield: 16.5 mg, 21%; Appearance: White solid;1H NMR (400 MHz, DMSO-d6) δ 11.34 (s, 1H), 8.16 (dd, J = 4.8, 1.2 Hz, 1H), 7.99 – 7.85 (m, 3H), 7.26 (d, J = 2.4 Hz, 1H), 7.00 (dd, J = 7.6, 4.8 Hz, 1H), 4.08 (t, J = 8.4 Hz, 1H), 3.54 (dd, J = 7.6, 5.6 Hz, 1H), 3.03 (dd, J = 14.4, 5.2 Hz, 1H), 2.73 (dd, J = 14.4, 8.0 Hz, 1H), 2.55 (d, J = 4.8 Hz, 3H), 1.95 – 1.64 (m, 2H), 1.64 – 1.55 (m, 1H), 1.33 – 1.22 (m, 1H), 0.99 – 0.84 (m, 1H), 0.78 – 0.68 (m, 6H); HPLC purity: 100%; LCMS Calculated for C17H25N5O2: 331.20; Observed: 332.4 [M+H]+. and (2S,3S)-2-[(2R)-2-amino-3-{1H-pyrrolo[2,3- b]pyridin-3-yl}propanamido]-N,3-dimethylpentanamide (A-28). Yield: 39.8 mg, 52%; Appearance: White solid;1H NMR (400 MHz, DMSO-d6) δ 11.36 (s, 1H), 8.16 (dd, J = 4.8, 1.2 Hz, 1H), 8.00 – 7.84 (m, 3H), 7.28 (d, J = 2.8 Hz, 1H), 7.00 (dd, J = 7.6, 4.8 Hz, 1H), 4.07 (t, J = 8.0 Hz, 1H), 3.49 (dd, J = 7.6, 4.4 Hz, 1H), 3.04 (dd, J = 14.4, 4.4 Hz, 1H), 2.83 (dd, J = 14.4, 7.6 Hz, 1H), 2.56 (d, J = 4.8 Hz, 3H), 1.94 – 1.69 (m, 2H), 1.63 – 1.50 (m, 1H), 1.31 – 1.21 (m, 1H),0.97 – 0.80 (m, 1H), 0.74 (t, J = 7.2 Hz, 3H), 0.66 (d, J = 6.8 Hz, 3H); HPLC purity: 99.54%; LCMS Calculated for C17H25N5O2: 331.20; Observed: 332.4 [M+H]+.

[0439] The following examples were prepared using standard chemical manipulations and procedures similar to those used for the preparation of the previous examples. Analytical data is given in the table below.Example A6. Synthesis of (S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-((R)-1,4,4- trimethylpyrrolidin-3-yl)pentanamide (A-39):

[0440] Step-1. Synthesis of tert-butyl (R)-(1,4,4-trimethylpyrrolidin-3-yl)carbamate

[0441] To a solution of tert-butyl N-[(3R)-4,4-dimethylpyrrolidin-3-yl]carbamate (200mg, 933 µmol) in MeOH (5 mL) was added Paraformaldehyde (419 mg, 4.66 mmol), NaBH4(175 mg, 2.79 mmol) at rt. The reaction mixture was stirred for 2 h at 40 °C. The reaction mixture was added water (20 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated under reducedpressure to give tert-butyl N-[(3R)-1,4,4-trimethylpyrrolidin-3-yl]carbamate (150 mg, 656 µmol, 100% purity, 70% yield) as a colorless oil.

[0442] Step-2. Synthesis of (R)-1,4,4-trimethylpyrrolidin-3-amine

[0443] To a solution of tert-butyl N-[(3R)-1,4,4-trimethylpyrrolidin-3-yl]carbamate (150mg, 656 µmol) in DCM (5 mL) was added TFA (1 ml, 6.56 mmol) at rt. The reaction mixture was stirred for 1 h at rt. The reaction mixture was concentrated under reduced pressure to give (3R)- 1,4,4-trimethylpyrrolidin-3-amine (84.0 mg, 655 µmol, 100% purity, 100% yield) as a light-yellow oil.

[0444] Step-3. Synthesis of tert-butyl ((S)-5-(2-nitro-1H-imidazol-1-yl)-1-oxo-1-(((R)-1,4,4- trimethylpyrrolidin-3-yl)amino)pentan-2-yl)carbamate

[0445] To a solution of (3R)-1,4,4-trimethylpyrrolidin-3-amine (50mg, 389 µmol) in DMF (5 mL) was added (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H-imidazol-1-yl)pentanoic acid, INT-3 (127 mg, 389 µmol) , DEPBT (139 mg, 466 µmol) , DIPEA (50.5 mg, 389 µmol) at rt. The reaction mixture was stirred for 2 h at rt. The reaction mixture was added water (20 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by flash silica gel chromatography (MeOH / DCM = 0-10%) to give tert-butyl N-[(1S)-4-(2-nitro-1H-imidazol-1-yl)- 1-{[(3R)-1,4,4-trimethylpyrrolidin-3-yl]carbamoyl}butyl]carbamate (70.0 mg, 159 µmol, 100% purity, 41% yield) as a light yellow oil.

[0446] Step-4. tert-butyl ((S)-5-(2-amino-1H-imidazol-1-yl)-1-oxo-1-(((R)-1,4,4- trimethylpyrrolidin-3-yl)amino)pentan-2-yl)carbamate, A-39

[0447] To a solution of tert-butyl N-[(1S)-4-(2-nitro-1H-imidazol-1-yl)-1-{[(3R)-1,4,4- trimethylpyrrolidin-3-yl]carbamoyl}butyl]carbamate (100mg, 228 µmol) in MeOH (5 mL) was added Pd / C (36.2 mg, 342 µmol) at rt. The reaction mixture was stirred for 1 h at rt. The mixture was filtered through a celite pad, and the filtrate was concentrated under reduced pressure to give tert-butyl N-[(1S)-4-(2-amino-1H-imidazol-1-yl)-1-{[(3R)-1,4,4-trimethylpyrrolidin-3-yl] carbamoyl}butyl]carbamate (93.0 mg, 227 µmol, 100% purity, 99% yield) as a light yellow oil.

[0448] Step-5. Synthesis of (S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-((R)-1,4,4- trimethylpyrrolidin-3-yl)pentanamide

[0449] To a solution of tert-butyl N-[(1S)-4-(2-amino-1H-imidazol-1-yl)-1-{[(3R)-1,4,4- trimethylpyrrolidin-3-yl]carbamoyl}butyl]carbamate (93mg, 227 µmol) in DCM (5 mL) wasadded TFA (1 ml, 227 µmol) at rt. The reaction mixture was stirred for 1 h at rt. Then the mixture was concentrated. The residue was dissolved in MeOH and adjusted PH to 9 with NH3 / MeOH and then purified by pre-HPLC (0.1%NH4OH method) to give (2S)-2-amino-5-(2-amino-1H-imidazol- 1-yl)-N-[(3R)-1,4,4-trimethylpyrrolidin-3-yl]pentanamide (A-39). Yield: 23.0 mg, 32%; Appearance: White solid; 1H NMR (400 MHz, DMSO) δ 7.78 – 7.65 (m, 1H), 6.51 (d, J = 1.2 Hz, 1H), 6.33 (d, 1H), 5.31 (d, J = 32.0 Hz, 2H), 3.92 (dd, J = 15.6, 7.2 Hz, 1H), 3.63 (t, J = 7.2 Hz, 2H), 3.16 (dd, J = 7.6, 5.0 Hz, 1H), 2.84 – 2.75 (m, 1H), 2.31 (d, J = 8.8 Hz, 1H), 2.27 – 2.14 (m, 5H), 1.70 – 1.43 (m, 3H), 1.36 – 1.23 (m, 1H), 1.04 (s, 3H), 0.82 (d, J = 9.2 Hz, 3H); HPLC purity: 100%; LCMS Calculated for C15H28N6O: 308.43; Observed: 309.3 [M+H]+. Example A7. Synthesis of (S)-N-((R)-1-acetylpyrrolidin-3-yl)-2-amino-5-(2-amino-1H- imidazol-1-yl)pentanamide (A-40)

[0450] Step-1. Synthesis of tert-butyl (R)-(1-acetylpyrrolidin-3-yl)carbamate

[0451] To a solution of tert-butyl (R)-pyrrolidin-3-ylcarbamate (2 g, 10.7 mmol) in DCM (50 mL) was added acetyl chloride (1.67 g, 21.4 mmol) and triethylamine (2.16 g, 21.4 mmol) at 0 °C. The mixture was stirred at 0 °C for 2 h. The mixture was concentrated in vacuum. The residue waspurified by flash silica gel chromatography (MeOH / DCM = 0-4%) to afford tert-butyl (R)-(1- acetylpyrrolidin-3-yl)carbamate (2.4 g, 10.6 mmol, 100% purity, 99% yield) as a white solid.

[0452] Step-2. Synthesis of (R)-1-(3-aminopyrrolidin-1-yl)ethan-1-one hydrochloride

[0453] To a solution of tert-butyl (R)-(1-acetylpyrrolidin-3-yl)carbamate (2.4 g, 10.5 mmol) in 4 M HCl in MeOH (50 mL ) was stirred at RT for 2 h. The mixture was concentrated in vacuum to give (R)-1-(3-aminopyrrolidin-1-yl)ethan-1-one hydrochloride (1.72 g, 10.4 mmol, 100% purity, 100% yield) as a white solid. The crude was used for the next step directly without further purification.

[0454] Step-3. Synthesis of methyl (S)-5-(((R)-1-acetylpyrrolidin-3-yl)amino)-4-((tert- butoxycarbon-yl)amino)-5-oxopentanoate

[0455] To a solution of (S)-2-((tert-butoxycarbonyl)amino)-5-methoxy-5-oxopentanoic acid (2.58 g, 9.90 mmol) in DMF (30 mL ) was added (R)-1-(3-aminopyrrolidin-1-yl)ethan-1-one hydrochloride (1.63 g, 9.90 mmol), EDCI (2.64 g, 13.8 mmol), HOBT (1.86 g, 13.8 mmol) and DIPEA (6.39 g, 49.5 mmol) at RT. The mixture was stirred at RT for 10 h. The mixture was quenched with water, extracted with EA. The organic layers were collected, washed with water, concentrated in vacuum. The residue was purified by flash silica gel chromatography (MeOH / DCM = 0-4%) to give methyl (S)-5-(((R)-1-acetylpyrrolidin-3-yl)amino)-4-((tert- butoxycarbon-yl)amino)-5-oxopentanoate (1.50 g, 4.03 mmol, 100% purity, 41% yield) as the colorless oil.

[0456] Step-4. Synthesis of (S)-5-(((R)-1-acetylpyrrolidin-3-yl)amino)-4-((tert- butoxycarbonyl)amino)-5-oxopentanoic acid

[0457] To a solution of methyl (S)-5-(((R)-1-acetylpyrrolidin-3-yl)amino)-4-((tert- butoxycarbon-yl)amino)-5-oxopentanoate (1.50 g, 4.03 mmol) in THF (30 mL ) was added LiOH (106 mg, 4.43 mmol) and H2O (10 mL ) at RT. The mixture was stirred at RT for 10 h. The mixture was quenched with 1 M HCl, extracted with EA. The organic layers were concentrated in vacuum to give (S)-5-(((R)-1-acetylpyrrolidin-3-yl)amino)-4-((tert-butoxycarbonyl)amino)-5- oxopentanoic acid (1.30 g, 3.63 mmol, 100% purity, 90% yield) as a white solid. The crude was used for the next step directly without further purification.

[0458] Step-5. Synthesis of tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-hydroxy- 1-oxopent-an-2-yl)carbamate

[0459] To a stirred solution of (S)-5-(((R)-1-acetylpyrrolidin-3-yl)amino)-4-((tert- butoxycarbonyl)-amino)-5-oxopentanoic acid (500 mg, 1.39 mmol) in DCM (10 mL ) was added BH3(4 mL, 1M in THF ) at 0 °C. The mixture was stirred at 0 °C for 0.5 h. The mixture was quenched with MeOH, concentrated in vacuum. The residue was purified by flash silica gel chromatography (MeOH / DCM = 0-4%) to give tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3- yl)amino)-5-hydroxy-1-oxopent-an-2-yl)carbamate (125 mg, 363 µmol, 100% purity, 26% yield) as the colorless oil.

[0460] Step-6. Synthesis of tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-(2-nitro- 1H-imidaz-ol-1-yl)-1-oxopentan-2-yl)carbamate

[0461] A solution of tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-hydroxy-1- oxopent-an-2-yl)carbamate (125 mg, 363 µmol) in THF (8 mL ) was added to PPh3(190 mg, 726 µmol), 2-nitro-1H-imidazole (82 mg, 726 µmol) and DIAD (146 mg, 726 µmol) at RT. The mixture was stirred at RT for 12 h. The mixture was quenched with water, extracted with EA. The organic layers were concentrated in vacuum. The residue was purified by flash silica gel chromatography (MeOH / DCM = 0-4%) to give tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3- yl)amino)-5-(2-nitro-1H-imidaz-ol-1-yl)-1-oxopentan-2-yl)carbamate (85 mg, 193 µmol, 100% purity, 53% yield) as the colorless oil.

[0462] Step-7. Synthesis of tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-(2-amino- 1H-imidazol-1-yl)-1-oxopentan-2-yl)carbamate

[0463] To a solution of tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-(2-nitro-1H- imidaz-ol-1-yl)-1-oxopentan-2-yl)carbamate (85 mg, 193 µmol) in MeOH (30 mL ) was added 10% Pd / C (30 mg ) at RT. The mixture was stirred at RT for 16 h under H2atmosphere. The mixture was filtered, concentrated in vacuum to give tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3- yl)amino)-5-(2-amino-1H-imidazol-1-yl)-1-oxopentan-2-yl)carbamate (79 mg, 192 µmol, 100% purity, 100% yield) as the colorless oil. The crude was used for the next step directly without further purification.

[0464] Step-8. Synthesis of (S)-N-((R)-1-acetylpyrrolidin-3-yl)-2-amino-5-(2-amino-1H- imidazol-1-yl)pentanamide

[0465] To a solution of tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-(2-amino-1H- imidazol-1-yl)-1-oxopentan-2-yl)carbamate (79 mg, 193 µmol) in (20 mL, 4 M in dioxane) was stirred at RT for 1 h. The resulting mixture was concentrated. The residue was purified by prep-HPLC (ACN / water / 0.1% NH4OH) to give (S)-N-((R)-1-acetylpyrrolidin-3-yl)-2-amino-5-(2- amino-1H-imidazol-1-yl)pentanamide (A-40). Yield: 33.1 mg, 52%; Appearance: colorless oil;1H NMR (400 MHz, DMSO-d6) δ 8.05 (dd, J1 = 7.2, J2 = 6.8 Hz, 1H), 6.52 (d, J = 1.2 Hz, 1H), 6.34 (d, J = 1.2 Hz, 1H), 5.27 (s, 2H), 4.27 – 4.16 (m, 1H), 3.65 – 3.62 (m, 2H), 3.50 – 3.42 (m, 2H), 3.22 – 3.08 (m, 3H), 2.01 – 1.90 (m, 1H), 1.93 (d, J = 9.2 Hz, 3H), 1.86 – 1.70 (m, 1H), 1.67 – 1.41 (m, 3H), 1.30 – 1.26 (m, 1H); HPLC purity: 94.3 %; LCMS Calculated for C14H24N6O2: 308.39; Observed: 309.4 [M+H]+.

[0466] The following examples were prepared using standard chemical manipulations and procedures similar to those used for the preparation of the previous examples. Analytical data is given in the table below.Example A8. Synthesis of (2S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-[(1S)-1- (methylcarbamoyl)-2-(oxan-4-yl)ethyl]pentanamide (A-52):

[0467] Step-1. Synthesis of tert-butyl N-[(1S)-1-(methylcarbamoyl)-2-(oxan-4- yl)ethyl]carbamate

[0468] To a solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-(oxan-4-yl)propanoic acid (100 mg, 365 µmol), Methylamine hydrochloride (29.5 mg, 438 µmol), DIPEA (140 mg, 1.09 mmol) in DMF (5 mL) was added HATU (180 mg, 474 µmol) at rt. The reaction mixture was stirred for 2 h at rt. The reaction mixture was added water (20 mL) and extracted with EA (30 mL x 3). The organic layer was washed with water (20 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated to give residue, which was purified by flash silica gel chromatography (DCM / MeOH = 20 / 1) to give tert-butyl N-[(1S)-1-(methylcarbamoyl)-2-(oxan- 4-yl)ethyl]carbamate (92.0 mg, 321 µmol, 100% purity, 88% yield) as a light yellow oil.

[0469] Step-2. Synthesis of (2S)-2-amino-N-methyl-3-(oxan-4-yl)propanamide

[0470] A solution of tert-butyl N-[(1S)-1-(methylcarbamoyl)-2-(oxan-4-yl)ethyl]carbamate (92 mg, 321 µmol) in HCl in dioxane (4M) (5 mL) was stirred for 1 h at rt. The reaction mixture was concentrated to give (2S)-2-amino-N-methyl-3-(oxan-4-yl)propanamide (59.2 mg, 317 µmol, 100% purity, 99% yield) as a light yellow oil.

[0471] Step-3. Synthesis of tert-butyl N-[(1S)-1-{[(1S)-1-(methylcarbamoyl)-2-(oxan-4- yl)ethyl]carbamoyl}-4-(2-nitro-1H-imidazol-1-yl)butyl]carbamate

[0472] To a solution of (2S)-2-amino-N-methyl-3-(oxan-4-yl)propanamide (59 mg, 316 µmol), (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H-imidazol-1-yl)pentanoic acid, INT- 3 (124 mg, 379 µmol), DEPBT (113 mg, 379 µmol) in DMF (5 mL) was added DIPEA (122 mg, 948 µmol) at rt. The reaction mixture was stirred for 2 h at rt. The reaction mixture was added water (20 mL) and extracted with EA (30 mL X 3). The organic layer was washed with water (20 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated to give residue, which was purified by flash silica gel chromatography (DCM / MeOH = 20 / 1) to give tert-butyl N-[(1S)-1- {[(1S)-1-(methylcarbamoyl)-2-(oxan-4-yl)ethyl]carbamoyl}-4-(2-nitro-1H-imidazol-1- yl)butyl]carbamate (105 mg, 211 µmol, 100% purity, 67% yield) as a colorless oil.

[0473] Step-4. Synthesis of tert-butyl N-[(1S)-4-(2-amino-1H-imidazol-1-yl)-1-{[(1S)-1- (methylcarbamoyl)-2-(oxan-4-yl)ethyl]carbamoyl}butyl]carbamate

[0474] To a solution of tert-butyl N-[(1S)-1-{[(1S)-1-(methylcarbamoyl)-2-(oxan-4- yl)ethyl]carbamoyl}-4-(2-nitro-1H-imidazol-1-yl)butyl]carbamate (105 mg, 211 µmol) in MeOH (5 mL) was added Pd / C (10 mg) at rt. The reaction mixture was stirred for 1 h at rt under H2. The reaction mixture was filtered and concentrated to give tert-butyl N-[(1S)-4-(2-amino-1H-imidazol- 1-yl)-1-{[(1S)-1-(methylcarbamoyl)-2-(oxan-4-yl)ethyl]carbamoyl}butyl]carbamate (98.0 mg, 210 µmol, 100% purity, 100% yield) as a light yellow solid.

[0475] Step-5. Synthesis of (2S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-[(1S)-1- (methylcarbamoyl)-2-(oxan-4-yl)ethyl]pentanamide

[0476] To a solution of tert-butyl N-[(1S)-4-(2-amino-1H-imidazol-1-yl)-1-{[(1S)-1- (methylcarbamoyl)-2-(oxan-4-yl)ethyl]carbamoyl}butyl]carbamate (98 mg, 210 µmol) in DCM (5 mL) was added TFA (1 mL, 210 µmol) at rt. The reaction mixture was stirred for 1 h at rt. The reaction mixture was adjusted pH to 7-8 with NH3in MeOH and concentrated to give residue, which was further purified by Prep-HPLC (ACN / water / 0.1% FA) to give (2S)-2-amino-5-(2- amino-1H-imidazol-1-yl)-N-[(1S)-1-(methylcarbamoyl)-2-(oxan-4-yl)ethyl]pentanamide, (A-52).Yield: 7.8 mg, 9.2%; Appearance: White solid;1H NMR (400 MHz, DMSO-d6) δ 8.06 – 7.78 (m, 2H), 6.52 (d, J = 1.2 Hz, 1H), 6.34 (d, J = 1.2 Hz, 1H), 5.23 (s, 2H), 4.36 – 4.23 (m, 1H), 3.79 (dd, J = 11.2, 2.4 Hz, 2H), 3.63 (t, J = 7.2 Hz, 2H), 3.18 (tt, J = 11.6, 10.8 Hz, 3H), 2.56 (d, J = 4.6 Hz, 3H), 1.68 – 1.31 (m, 9H), 1.21 – 1.03 (m, 2H); HPLC purity: 82.6%; LCMS Calculated for C17H30N6O3: 366.24; Observed: 367.4 [M+H]+. Example A9. Synthesis of (S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-2-methyl-N-((2S,3S)-3- methyl-1-(methylamino)-1-oxopentan-2-yl)pentanamide, (A-53).

[0477] Step-1. Synthesis of (9H-fluoren-9-yl)methyl ((2S,3S)-3-methyl-1-(methylamino)-1- oxopentan-2-yl)carbamate

[0478] To a solution of (2S,3S)-2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)-3- methylpentanoic acid (300 mg, 848 µmol) in DMF (3 mL) was added DEPBT (301 mg, 1.01mmol), DIEA (735 µL, 4.23 mmol) and methanamine hydrochloride (68.1 mg, 1.01 mmol). The mixture was stirred at rt for 3 h. Then water was added to the mixture and the precipitate was collected by filtration to give (9H-fluoren-9-yl)methyl ((2S,3S)-3-methyl-1-(methylamino)-1- oxopentan-2-yl)carbamate (228 mg, 622 µmol, 100% purity, 73.5% yield) as a light yellow solid

[0479] Step-2. Synthesis of (2S,3S)-2-amino-N,3-dimethylpentanamide

[0480] A mixture of (9H-fluoren-9-yl)methyl ((2S,3S)-3-methyl-1-(methylamino)-1- oxopentan-2-yl)carbamate (228 mg, 622 µmol) and dimethylamine (3 mL, 622 µmol) in THF / MeOH (3 mL) was stirred at rt overnight. Then the mixture was concentrated. The residue was dissolved with water (10 mL) and washed with EtOAc (2 x 10 mL). The aqueous phase was concentrated to give (2S,3S)-2-amino-N, 3-dimethylpentanamide (88.8 mg, 615 µmol, 100% purity, 99% yield) as colorless oil.

[0481] Step-3. Synthesis of tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2- methylpent-4-enoate

[0482] To a solution of (2S)-2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)-2-methylpent- 4-enoic acid (1 g, 2.84 mmol) in DCM (40 mL) was added tert-butyl 2,2,2- trichloroethanimidate (2.46 g, 11.3 mmol). The mixture was stirred at 35 °C for 3 days. Then the mixture was concentrated and the residue was purified by silica gel column (PE: EtOAc= 30: 1) to give tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methylpent-4-enoate (980 mg, 2.40 mmol, 100% purity, 85% yield) as colorless oil.

[0483] Step-4. Synthesis of tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5- hydroxy-2-methylpentanoate

[0484] To a solution of tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2- methylpent-4-enoate 190 mg, 466 µmol) in THF (10 mL) at 0 °C, was added 9-BBN (3.72 mL, 1.86 mmol). The mixture was stirred at rt overnight. Then the mixture was cooled to 0 °C, and quenched with water. Then a solution of NaOAc (633 mg, 4.66 mmol) in H2O (2 mL) and H2O2(2 mL, 25.7 mmol) was added dropwise. The mixture was stirred at rt for 1 h. Then the mixture was diluted with EtOAc (35 mL) and washed with brine (3 x 30 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography (PE / EtOAc = 2 / 1) to give tert-butyl (S)-2-((((9H-fluoren-9- yl)methoxy)carbonyl)amino)-5-hydroxy-2-methylpentanoate (173 mg, 406 µmol, 100% purity, 87% yield) as colorless oil.

[0485] Step-5. Synthesis of tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2- methyl-5-(2-nitro-1H-imidazol-1-yl)pentanoate

[0486] At 0 °C, to a solution of tert-butyl (S)-2-((((9H-fluoren-9- yl)methoxy)carbonyl)amino)-5-hydroxy-2-methylpentanoate (173 mg, 406 µmol), 2-nitro-1H- imidazole (55.0 mg, 487 µmol) and PPh3 (170 mg, 649 µmol) in THF (2 mL) was added DIAD (127 µL, 649 µmol). The mixture was stirred at rt overnight. Then the mixture was concentrated. The residue was purified by silica gel column (PE: EtOAc = 2: 1) to give tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methyl-5-(2-nitro-1H-imidazol-1- yl)pentanoate (170 mg, 326 µmol, 100% purity, 80.5% yield) as a white solid.

[0487] Step-6. Synthesis of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methyl-5- (2-nitro-1H-imidazol-1-yl)pentanoic acid

[0488] To a solution of tert-butyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2- methyl-5-(2-nitro-1H-imidazol-1-yl)pentanoate (170 mg, 326 µmol) in DCM (1.5 mL) was added TFA (0.5 mL, 6.72 mmol). The mixture was stirred at rt for 2 h. Then the mixture was concentrated to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methyl-5-(2-nitro-1H-imidazol-1- yl)pentanoic acid (150 mg, 322 µmol, 100% purity, 99% yield) as a white solid, which was used directly in the nest step without purification.

[0489] Step-7. Synthesis of (9H-fluoren-9-yl)methyl ((S)-2-methyl-1-(((2S,3S)-3-methyl-1- (methylamino)-1-oxopentan-2-yl)amino)-5-(2-nitro-1H-imidazol-1-yl)-1-oxopentan-2- yl)carbamate

[0490] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-methyl-5-(2- nitro-1H-imidazol-1-yl)pentanoic acid (90 mg, 193 µmol) in DMF (3 mL) were added DEPBT (69.0 mg, 231 µmol), DIEA (100 µL, 579 µmol) and (2S,3S)-2-amino-N,3-dimethylpentanamide (139 mg, 965 µmol). The mixture was stirred at rt overnight. Then the mixture was diluted with EtOAc (30 mL) and washed with brine (3 x 20 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography (DCM / MeOH = 20 / 1) to give (9H-fluoren-9-yl)methyl ((S)-2-methyl-1-(((2S,3S)-3-methyl-1- (methylamino)-1-oxopentan-2-yl)amino)-5-(2-nitro-1H-imidazol-1-yl)-1-oxopentan-2- yl)carbamate (37.0 mg, 62.6 µmol, 100% purity, 32% yield) as an off-white solid.

[0491] Step-8. Synthesis of (9H-fluoren-9-yl)methyl ((S)-5-(2-amino-1H-imidazol-1-yl)-2- methyl-1-(((2S,3S)-3-methyl-1-(methylamino)-1-oxopentan-2-yl)amino)-1-oxopentan-2- yl)carbamate

[0492] To a solution of (9H-fluoren-9-yl)methyl ((S)-2-methyl-1-(((2S,3S)-3-methyl-1- (methylamino)-1-oxopentan-2-yl)amino)-5-(2-nitro-1H-imidazol-1-yl)-1-oxopentan-2- yl)carbamate (37 mg, 62.6 µmol) in MeOH (3 mL) was added Pd / C (6.63 mg, 62.6 µmol). The mixture was stirred at rt for 1 h under H2(1 atm). Then the mixture was filtered. The filtrate was concentrated to give (9H-fluoren-9-yl)methyl ((S)-5-(2-amino-1H-imidazol-1-yl)-2-methyl-1- (((2S,3S)-3-methyl-1-(methylamino)-1-oxopentan-2-yl)amino)-1-oxopentan-2-yl)carbamate (32.0 mg, 57.0 µmol, 100% purity, 91% yield) as a light brown solid.

[0493] Step-9. Synthesis of (S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-2-methyl-N-((2S,3S)- 3-methyl-1-(methylamino)-1-oxopentan-2-yl)pentanamide (A-53)

[0494] A solution of (9H-fluoren-9-yl)methyl ((S)-5-(2-amino-1H-imidazol-1-yl)-2-methyl-1- (((2S,3S)-3-methyl-1-(methylamino)-1-oxopentan-2-yl)amino)-1-oxopentan-2-yl)carbamate (32 mg, 57.0 µmol) in dimethylamine (3 mL, 57.0 µmol) was stirred at rt for 1 h. Then the mixture was concentrated. The residue was purified by silica gel column (DCM: MeOH: NH3= 30: 1: 0.1) to give the crude product, which was further purified by pre-HPLC (ACN / water / 0.1% NH4HCO3) to give (S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-2-methyl-N-((2S,3S)-3-methyl-1- (methylamino) -1-oxopentan-2-yl)pentanamide, (A-53). Yield: 12.0 mg, 62.5%; Appearance: Brown oil; 1H NMR (400 MHz, DMSO-d6) δ 8.06-7.94 (m, 2H), 6.49 (d, J = 1.4 Hz, 1H), 6.33 (d, J = 1.4 Hz, 1H), 5.20 (s, 2H), 4.10-4.06 (m, 1H), 3.59 (t, J = 7.0 Hz, 2H), 2.58 (d, J = 4.6 Hz, 3H), 2.10-1.90 (m, 2H), 1.74 – 1.55 (m, 3H), 1.52 – 1.32 (m, 3H), 1.12 (s, 3H), 1.07 – 0.94 (m, 1H), 0.85 – 0.72 (m, 6H); HPLC purity: 100%; LCMS Calculated for C16H30N6O2: 338.24; Observed: 339.2 [M+H]+. Example A10. Synthesis of (S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-((R)-1-(4-(4- methylpiperazin-1-yl)-4-oxobutyl)pyrrolidin-3-yl)pentanamide, (A-54)

[0495] Step-1. Synthesis of tert-butyl ((S)-1-(((R)-1-(4-(4-methylpiperazin-1-yl)-4- oxobutyl)pyrrolidin-3-yl)amino)-5-(2-nitro-1H-imidazol-1-yl)-1-oxopentan-2-yl)carbamate

[0496] To a solution of 4-((R)-3-((S)-2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H-imidazol- 1-yl)pentanamido)pyrrolidin-1-yl)butanoic acid (60 mg, 124 µmol), 1-methylpiperazine (25 mg, 248 µmol), 1-[(dimethylamino)(dimethyliminio)methyl]-3-oxo-2H,3H-3lambda5-[1,2,3]triazolo [5,4-b]pyridin-3-ylium-2-ide; hexafluoro-lambda5-phosphanuide (94 mg, 248 µmol) and N,N- diisopropylethylamine (48 mg, 372 µmol) in DMF (3 mL ) was stirred in RT for 3 h. The reaction mixture was added water and extracted with ethyl acetate (3 mL x 3). The organic layer was washed with water (2 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated to give residue, which was purified by flash chromatography (MeOH / DCM = 0-6%) to give tert-butyl ((S)-1-(((R)-1-(4-(4-methylpiperazin-1-yl)-4-oxobutyl)pyrrolidin-3-yl)amino)-5-(2-nitro-1H- imidazol-1-yl)-1-oxopentan-2-yl)carbamate (66 mg, 116 µmol, 94% purity, 94% yield) as the colorless oil.

[0497] Step-2. Synthesis of tert-butyl ((S)-5-(2-amino-1H-imidazol-1-yl)-1-(((R)-1-(4-(4- methylpiperazin-1-yl)-4-oxobutyl)pyrrolidin-3-yl)amino)-1-oxopentan-2-yl)carbamate

[0498] To a solution of tert-butyl ((S)-1-(((R)-1-(4-(4-methylpiperazin-1-yl)-4- oxobutyl)pyrrolidin-3-yl)amino)-5-(2-nitro-1H-imidazol-1-yl)-1-oxopentan-2-yl)carbamate (46 mg, 81 µmol) in MeOH (3 mL ) was added 10% Pd / C (14 mg) at RT. The reaction mixture was stirred for 2 h under H2. The reaction mixture was filtered, concentrated to give tert-butyl ((S)-5- (2-amino-1H-imidazol-1-yl)-1-(((R)-1-(4-(4-methylpiperazin-1-yl)-4-oxobutyl)pyrrolidin-3- yl)amino)-1-oxopentan-2-yl)carbamate (58 mg, 108 µmol, 90% purity, 96% yield) as a yellow solid, which was used for next step directly without further purification.

[0499] Step-3. Synthesis of (S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-((R)-1-(4-(4- methylpiperazin-1-yl)-4-oxobutyl)pyrrolidin-3-yl)pentanamide

[0500] To a solution of tert-butyl ((S)-5-(2-amino-1H-imidazol-1-yl)-1-(((R)-1-(4-(4- methylpiperazin-1-yl)-4-oxobutyl)pyrrolidin-3-yl)amino)-1-oxopentan-2-yl)carbamate (58 mg, 108 µmol) in DCM (3 mL ) was added TFA (1 mL ) at rt. The mixture was stirred at rt for 1 h. The mixture was concentrated in vacuum to give crude product. The residue was purified by prep- HPLC (ACN / water / 0.1% NH3.H2O) to give(S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-((R)-1- (4-(4-methylpiperazin-1-yl)-4-oxobutyl)pyrrolidin-3-yl)pentanamide (A-54). Yield: 33.4 mg, 71%; Appearance: White solid;1H NMR (400 MHz, DMSO-d6) δ 7.60 (s, 1H), 6.49 (d, J = 1.6 Hz, 1H), 6.34 (d, J = 1.6 Hz, 1H), 4.88 (s, 2H), 4.24 – 4.09 (m, 1H), 3.64 (t, J = 7.2 Hz, 2H), 3.42 (t, J = 5.2 Hz, 4H), 3.14 – 3.09 (m, 1H), 2.66 – 2.59 (m, 2H), 2.41 – 2.24 (m, 10H), 2.18 (s, 3H), 2.11 – 2.02 (m, 1H), 1.69 – 1.53 (m, 6H), 1.41 – 1.29 (m, 1H); HPLC purity: 100.0%; LCMS Calculated for C28H30N4O2S: 434.59; Observed: 435.3 [M+H]+. Example A11. Synthesis of (S)-N-((R)-1-acetylpyrrolidin-3-yl)-2-amino-5-(2-amino-1H- benzo[d]imidazol-1-yl)pentanamide (A-55):

[0501] Step-1. Synthesis of tert-butyl (R)-(1-acetylpyrrolidin-3-yl)carbamate

[0502] To a solution of tert-butyl N-[(3R)-pyrrolidin-3-yl]carbamate (300 mg, 1.61 mmol) in DCM (3 mL ) was added acetyl chloride (252 mg, 3.22 mmol), TEA (325 mg, 3.22 mmol) at 0 °C. The resulting mixture was stirred at 0 °C for 3 h. The mixture was quenched with water and the aqueous layer was extracted with DCM (50 mL x 3). The organic layers were concentrated. The residue was purified by flash chromatography (MeOH / DCM = 0-6%) to give tert-butyl (R)-(1- acetylpyrrolidin-3-yl)carbamate (265 mg, 1.16mmol, 100% purity, 72% yield) as a white solid.

[0503] Step-2. Synthesis of (R)-1-(3-aminopyrrolidin-1-yl)ethan-1-one

[0504] To a solution of tert-butyl (R)-(1-acetylpyrrolidin-3-yl)carbamate (175 mg, 766 μmol) in DCM (4 mL ) was added TFA (2 mL ). The resulting mixture was stirred at RT for 1 h. The reaction mixture was concentrated to give the crude (R)-1-(3-aminopyrrolidin-1-yl)ethan-1-one (98 g, 432 μmol, 100% purity, 56% yield) as a yellow oil.

[0505] Step-3. Synthesis of tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-(2-nitro- 1H-benzo[d]imidazol-1-yl)-1-oxopentan-2-yl)carbamate

[0506] To a solution of (R)-1-(3-aminopyrrolidin-1-yl)ethan-1-one (50 mg, 390 µmol) in DMF (2 mL ) was added (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H-1,3-benzodiazol-1- yl)pentanoic acid (147 mg, 390 μmol), DEPBT (175 mg, 585 μmol) and DIPEA (252 mg, 1.95 mmol) . The resulting mixture was stirred at RT for 16 h. The mixture was diluted with water, extracted with EA (60 mL x 3). The organic layers were concentrated. The residue was purified by flash chromatography (MeOH / DCM = 0-6%) to give tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin- 3-yl)amino)-5-(2-nitro-1H-benzo[d]imidazol-1-yl)-1-oxopentan-2-yl)carbamate (100 mg, 204 μmol, 100% purity, 53% yield) as the green oil.

[0507] Step-4. Synthesis of tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-(2-amino- 1H-benzo[d]imidazol-1-yl)-1-oxopentan-2-yl)carbamate

[0508] To a solution of tert-butyl (S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-(2-nitro-1H- benzo[d]imidazol-1-yl)-1-oxopentan-2-yl)carbamate (100mg, 204 μmol) in MeOH (3 mL ) was added 10% Pd / C (33 mg). The reaction mixture was stirred for 2 h under H2. The reaction mixture was filtered, concentrated to give the crude tert-butyl ((S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)- 5-(2-amino-1H-benzo[d]imidazol-1-yl)-1-oxopentan-2-yl)carbamate (94 mg, 204 μmol) as a green oil, which was used for next step directly without further purification.

[0509] Step-5. Synthesis of (S)-N-((R)-1-acetylpyrrolidin-3-yl)-2-amino-5-(2-amino-1H- benzo[d]imi-dazol-1-yl)pentanamide

[0510] To a solution of tert-butyl (S)-1-(((R)-1-acetylpyrrolidin-3-yl)amino)-5-(2-amino-1H- benzo[d]imidazol-1-yl)-1-oxopentan-2-yl)carbamate (94 mg, 204 μmol) in DCM (2 mL ) was added TFA (1 mL ). The resulting mixture was stirred at RT for 1 h. The mixture was concentrated in vacuum. The residue was purified by prep-HPLC (ACN / water / 0.1% FA) to give (S)-N-((R)-1- acetylpyrrolidin-3-yl)-2-amino-5-(2-amino-1H-benzo[d]imidazol-1-yl)pentanamide, (A-55). Yield: 22.9 mg, 31%; Appearance: colorless oil; 1H NMR (400 MHz, DMSO-d6) δ 8.58 (dd, J = 13.2, 6.4 Hz, 1H), 8.32 (s, 2H), 7.16 (t, J = 7.6 Hz, 2H), 6.95 (dt, J = 20.8, 7.2 Hz, 4H), 4.22 (dd,J = 30.8, 4.4 Hz, 1H), 4.02 (s, 2H), 3.51 (ddt, J = 17.2, 11.0, 5.6 Hz, 3H), 3.31 (dd, J = 11.2, 5.2 Hz, 1H), 3.20 (ddd, J = 12.4, 7.6, 3.6 Hz, 1H), 2.01 (ddd, J = 25.6, 13.2, 7.6 Hz, 1H), 1.90 (s, 3H), 1.76 – 1.63 (m, 5H); HPLC purity: 98.2%; LCMS Calculated for C18H26N6O2: 358.45; Observed: 359.2 [M+H]+.

[0511] The following examples were prepared using standard chemical manipulations and procedures similar to those used for the preparation of the previous examples. Analytical data is given in the table below.Example A12. Synthesis of (S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-((S)-1-(5- (methoxymethyl)-1,3,4-oxadiazol-2-yl)-2,2-dimethylpropyl)pentanamide, (A-57)

[0512] Step 1. Synthesis of tert-butyl (S)-(1-hydrazineyl-3,3-dimethyl-1-oxobutan-2- yl)carbamate

[0513] To a solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-3,3-dimethylbutanoic acid (2 g, 8.64 mmol) in THF (20 mL) was added carbonyldiimidazole (1.54 g, 9.50 mmol) and the reaction mixture was stirred at room temperature under nitrogen atmosphere. After 1.5 h, hydrazine hydrate (1.26 mL, 25.9 mmol) was added, and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated, and the residue was diluted withethyl acetate (100 ml), washed with water (2 x 50 ml). The organic layer was dried over sodium sulphate, filtered, and concentrated under reduced pressure to afford the desired intermediate (2.00 g, 8.15 mmol, 94.7% yield, 90% purity) as a colourless gum.

[0514] Step 2. Synthesis of tert-butyl (S)-(1-(2-(2-methoxyacetyl)hydrazineyl)-3,3-dimethyl- 1-oxobutan-2-yl)carbamate

[0515] To a solution of tert-butyl N-[(1S)-1-(hydrazinecarbonyl)-2,2- dimethylpropyl]carbamate (1.5 g, 6.11 mmol) in THF (10 ml) was added sodium bicarbonate (589 mg, 7.02 mmol) and methoxyacetyl chloride (695 mg, 6.41 mmol) at 0 ° C under nitrogen atmosphere in a single neck round bottom flask and the reaction mixture was slowly warmed to rt for 30 min. The reaction mixture was concentrated, and the residue was dissolved in ethyl acetate (100 mL) and water (30 mL). The organic layer was separated, dried over sodium sulphate, filtered, and concentrated under reduced pressure, then the obtained 2.5 g of crude compound was purified by flash chromatography with the elution of 45 % ethyl acetate in pet-ether which afforded the title compound (2.00 g, 6.30 mmol, 103% yield, 96% purity) as a colorless gum.

[0516] Step 3. Synthesis of tert-butyl (S)-(1-(5-(methoxymethyl)-1,3,4-oxadiazol-2-yl)-2,2- dimethylpropyl)carbamate

[0517] To a stirred solution of triphenylphosphine (2.96 g, 11.3 mmol) and iodine (2.86 g, 11.3 mmol) in DCM (5 ml) was added a solution of tert-butyl N-[(1S)-1-[N (1.8 g, 5.67 mmol) tert- butyl (S)-(1-(2-(2-methoxyacetyl)hydrazineyl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate (1.8 g, 5.67 mmol) and triethylamine (2.40 g, 23.8 mmol) in DCM (5 mL) at RT. The reaction mixture was stirred at RT under nitrogen atmosphere for 2 h. The reaction mixture was diluted with DCM (100 mL), washed with saturated sodium thiosulphate solution (2 x 50 mL) and water (50 mL). The organic layer was dried over sodium sulphate, filtered and concentrated under reduced pressure. The obtained crude was purified via flash chromatography and the product was isolated with the elution of 20 % ethyl acetate in pet-ether to afford the title compound (1.18 g, 3.94 mmol, yield: 69.8%, 95% purity) as a pale brown gum.

[0518] Step 4. Syntheis of (S)-1-(5-(methoxymethyl)-1,3,4-oxadiazol-2-yl)-2,2- dimethylpropan-1-amine

[0519] To a solution of tert-butyl N-[(1S)-1-[5-(methoxymethyl)-1,3,4-oxadiazol-2-yl]-2,2- dimethylpropyl]carbamate (1.18 g, 3.94 mmol) in DCM (10 mL) was added trifluoroacetic acid (4.49 g, 39.4 mmol) at 0 °C and the reaction mixture was stirred at RT for 19 h (Note: After 16 h,5 eqiuiv. of TFA was added as the reaction was not completed). The reaction mixture was concentrated under reduced pressure and the residue was basified with saturated sodium bicarbonate solution to pH~8 and extracted with DCM (100 mL). The organic layer was washed with water (10 mL) and brine (10 ml), dried over sodium sulphate, filtered and concentrated under reduced pressure to afford (1S)-1-[5-(methoxymethyl)-1,3,4-oxadiazol-2-yl]-2,2-dimethylpropan- 1-amine (630 mg, 3.16 mmol, 80.2% yield, 99% purity) as a brown oil.

[0520] Step 5. Synthesis of tert-butyl ((S)-1-(((S)-1-(5-(methoxymethyl)-1,3,4-oxadiazol-2- yl)-2,2-dimethylpropyl)amino)-5-(2-nitro-1H-imidazol-1-yl)-1-oxopentan-2-yl)carbamate

[0521] To a solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H-imidazol-1- yl)pentanoic acid (1.14 g, 3.49 mmol) and (1S)-1-[5- (methoxymethyl)-1,3,4-oxadiazol-2-yl]-2,2- dimethylpropan-1-amine (0.58 g, 2.91 mmol) in Ethyl acetate (1.5 mL) was added triethylamine (883 mg, 8.73 mmol) and T3P (2.76 g, 4.36 mmol) at 0 °C and the reaction mixture was stirred at RT in a single neck RB under nitrogen atmosphere for 3 h. The reaction mixture was diluted with ethyl acetate (100 mL), washed with water (2x50 mL), 10 % sodium bicarbonate solution (50 mL) and brine solution (25 mL). The organic layer was dried over sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl N-[(1S)-1-{[(1S)-1-[5- (methoxymethyl)- 1,3,4-oxadiazol-2-yl]-2,2-dimethylpropyl]carbamoyl}-4-(2-nitro-1H-imidazol-1- yl)butyl]carbamate (1.40 g, 2.74 mmol, 94.5% yield, 96% purity) as a sticky yellow solid.

[0522] Step 6. Synthesis of tert-butyl ((S)-5-(2-amino-1H-imidazol-1-yl)-1-(((S)-1-(5- (methoxymethyl)-1,3,4-oxadiazol-2-yl)-2,2-dimethylpropyl)amino)-1-oxopentan-2-yl)carbamate

[0523] To a solution of tert-butyl N-[(1S)-1-{[(1S)-1-[5-(methoxymethyl)-1,3,4-oxadiazol-2- yl]-2,2-dimethylpropyl]carbamoyl}-4-(2-nitro-1Himidazol-1-yl)butyl]carbamate (1.3 g, 2.55 mmol) in methanol (5 mL) was purged with nitrogen for 5 min then palladium (271 mg, 2.55 mmol) (10 % on carbon) was added and the reaction mixture was stirred at RT under hydrogen atmosphere for 8 h. The reaction mixture was passed through celite bed, washed with methanol (250 ml) and the filtrate was concentrated to afford tert-butyl N-[(1S)-4-(2- amino-1H-imidazol- 1-yl)-1-{[(1S)-1-[5-(methoxymethyl)-1,3,4-oxadiazol-2-yl]-2,2- dimethylpropyl]carbamoyl}butyl]carbamate (1.20 g, 2.50 mmol, 98.3% yield, 94% purity) as a white solid.

[0524] Step 7. Syntheis of (S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-((S)-1-(5- (methoxymethyl)-1,3,4-oxadiazol-2-yl)-2,2-dimethylpropyl)pentanamide

[0525] To a solution of tert-butyl N-[(1S)-4-(2-amino-1H-imidazol-1-yl)-1-{[(1S)-1-[5- (methoxymethyl)-1,3,4-oxadiazol-2-yl]-2,2- dimethylpropyl]carbamoyl}butyl]carbamate (0.2 g, 416 µmol) in DCM (3 mL) was added trifluoroacetic acid (327 µL, 2.07 mmol) at 0 °C and the reaction mixture was stirred at room temperature under nitrogen atmosphere. After 3 h, an aliquot was submitted for analysis. The reaction mixture is concentrated under reduced pressure to afford the crude product as a brown gum which was then purified by prep. HPLC purification. The collected prep HPLC fraction was freeze dried for 16 h to afford 0.15 g (95% yield) of (S)-2- amino-5-(2-amino-1H-imidazol-1-yl)-N-((S)-1-(5-(methoxymethyl)-1,3,4-oxadiazol-2-yl)-2,2- dimethylpropyl)pentanamide, (A-57). Appearance: White solid.1H NMR (400 MHz, DMSO-d6) δ 8.977 (d, J = 8 Hz, 1H), 7.80-8.5 (br s, 2H), 6.944 (d, J= 2.4 Hz, 1H), 6.891 (d, J = 2.4 Hz, 1H), 4.988 (d, J = 8 Hz, 1H), 4.599-4.680 (m, 2H), 3.997 (br s, 1H), 3.812 (br s, 2H), 3.316-3.336 (br s, 3H), 1.594-1.646 (m, 4H), 1.004 (s, 9H); LCMS purity: 98.59% (M+H = 380.2; HPLC purity: 95.65%; LCMS calculated for C17H29N7O3: 379.47; Observed: 380.2 [M+H]+. Example A13. Synthesis of (2S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-[(1S)-1-[5- (methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2-dimethylpropyl]pentanamide hydrochloride salt (A- 58)

[0526] Step-1. Synthesis of (2S)-2-{[(benzyloxy)carbonyl]amino}-3,3-dimethylbutanoic acid

[0527] To a turbid solution of (2S)-2-amino-3,3-dimethylbutanoic acid (1 g, 7.62 mmol) in tetrahydrofuran (14 mL) and water (6 mL) was added sodium bicarbonate (1.59 g, 19.0 mmol) in one lot at 0 °C. The resulting mixture was stirred at RT for 20 mins followed by the dropwise addition of benzyl chloroformate (3.88 g, 11.4 mmol) at 0 °C. After the addition, reaction mixture was stirred at RT for 16 hours. After 16 hours, reaction mixture was concentrated under reduced pressure to remove THF. The residual layer was cooled to 0 °C, acidified to pH =2 using 1.5N HCl carefully, extracted with EtOAc (3 x 50 mL). The combined organic layer was washed with water (1 x 20 mL) dried over sodium sulfate, filtered through a cotton plug and concentrated under reduced pressure to get the crude which was purified by silica gel column chromatography. Fractions were collected by eluting with 18% EtOAc in Pet Ether. Appropriate fractions were combined together and concentrated under reduced pressure to afford (2S)-2-{[(benzyloxy) carbonyl]amino}-3,3-dimethylbutanoic acid (1.12 g, 4.25 mmol, 55% yield, 94% purity) as a white solid.

[0528] Step-2. Synthesis of benzyl N-[(1S)-1-carbamoyl-2,2-dimethylpropyl]carbamate

[0529] To an ice cooled solution of (2S)-2-{[(benzyloxy)carbonyl]amino}-3,3- dimethylbutanoic acid (13.4 g, 36.8 mmol) in dimethylformamide (250 mL ) were added ammonium chloride (19.6 g, 368 mmol), hydroxybenzotriazole (7.44 g, 55.1 mmol), EDC-HCl (10.5 g, 55.1 mmol) and N,N-diisopropylethylamine (25.5 mL, 147 mmol). The resulting mixture was stirred at RT for 16 hours. After 16 hours reaction mixture was diluted with water (400 mL), extracted with EtOAc (2 x 500 mL). The combined organic layer was washed with water (1 x 250 mL), brine (1x 200 mL), dried over sodium sulfate, filtered through a cotton plug and concentrated under reduced pressure to obtain the crude which was purified by silica gel column chromatography. Fractions were collected by eluting with 30% EtOAc in petroleum ether. Appropriate fractions were combined together and concentrated under reduced pressure to afford benzyl N-[(1S)-1-carbamoyl-2,2-dimethylpropyl]carbamate (8.06 g, 30.4 mmol, 82% yield, 97% purity) as a white solid.

[0530] Step-3. Synthesis of benzyl N-[(1S)-1-cyano-2,2-dimethylpropyl]carbamate

[0531] To an ice cooled solution of benzyl N-[(1S)-1-carbamoyl-2,2- dimethylpropyl]carbamate (9.95 g, 37.2 mmol) in methylene chloride (200 mL) was added triethylamine (20.5 mL, 148 mmol) in one lot followed by the dropwise addition of phosphorus oxychloride (4.16 mL, 44.6 mmol). The resulting mixture was stirred at RT for 16 hours. After 16 hours reaction mixture was concentrated under reduced pressure to get the brown semi solid which was quenched with ice water (150 mL), extracted with EtOAc (2 x 200 mL). Combined organic layer was washed with aq.10% NaHCO3 (1 x 100 mL), water (1 x 100 mL), brine (1 x 100 mL), dried over sodium sulfate, filtered through a cotton plug and concentrated under reduced pressure to get the crude benzyl N-[(1S)-1-cyano-2,2-dimethylpropyl]carbamate (9.00 g, 36.5 mmol, 99% purity, 98% yield) as a brown liquid.

[0532] Step-4. Synthesis of benzyl N-[(1S)-1-[(Z)-N'-hydroxycarbamimidoyl]-2,2- dimethylpropyl]carbamate

[0533] To solution of benzyl N-[(1S)-1-cyano-2,2-dimethylpropyl]carbamate (9.1 g, 36.5 mmol) in ethyl alcohol (150 mL) was added hydroxylamine (50 wt. % in H2O) (17.1 mL, 292 mmol) in one lot. The resulting mixture was stirred at 80 °C for 3 hours. After 3 hours reaction mixture was concentrated under reduced pressure, co-evaporated with toluene (2x 100 mL) to get the crude benzyl N-[(1S)-1-[(Z)-N'-hydroxycarbamimidoyl]-2,2-dimethylpropyl]carbamate (17.8 g, 64.0 mmol, crude, 89% purity).

[0534] Step-5. Synthesis of benzyl N-[(1S)-1-[5-(methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2- dimethylpropyl]carbamate

[0535] To a solution of benzyl N-[(1S)-1-[(Z)-N (20, 63.6 mmol) in dimethylformamide (80 mL ) were added pyridine (12.78 mL, 159 mmol) and 4-dimethylaminopyridine (776 mg, 6.36 mmol) in one lot. The resulting reaction mixture was cooled to 0 °C, and added 2-methoxyacetyl chloride (8.7 mL, 95.4 mmol) dropwise. After the addition reaction mixture was stirred at RT for 4 hours then heated to 100 °C for 16 hours. After 16 hours the reaction mixture was cooled to RT, diluted with water (600 mL), extracted with EtOAc (2 x 500 mL). Combined organic layer was washed with 1.5N HCl (1x 100 mL), water (1 x 100 mL), brine (1x 150 mL), dried over sodium sulfate, filtered through a cotton plug and concentrated under reduced pressure to get the crude which was purified by silica gel column chromatography. Fractions were collected by eluting with 25% EtOAc in Pet Ether. Appropriate fractions were combined and concentrated under reduced pressure to afford benzyl N-[(1S)-1-[5-(methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2- dimethylpropyl] carbamate (5.00 g, 7.5 mmol, 24% yield, 87% purity) as a white solid.

[0536] Step-6. Synthesis of (1S)-1-[5-(methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2- dimethylpropan-1-amine hydrochloride

[0537] A solution of benzyl N-[(1S)-1-[5-(methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2- dimethylpropyl] carbamate (1.5 g, 3.89 mmol) in conc. HCl (30 mL, 970 mmol) was heated to 100 °C for 16 hours. After 16 hours reaction mixture was concentrated under reduced pressure, co- evaporated with toluene to afford (1S)-1-[5-(methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2- dimethylpropan-1-amine hydrochloride (610 mg, 2.58 mmol, 66% yield, 61% purity) as a pale brown liquid.

[0538] Step-7. Synthesis of tert-butyl N-[(1S)-1-{[(1S)-1-[5-(methoxymethyl)-1,2,4- oxadiazol-3-yl]-2,2-dimethylpropyl]carbamoyl}-4-(2-nitro-1H-imidazol-1-yl)butyl]carbamate

[0539] To an ice cooled solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H- imidazol-1-yl)pentanoic acid (1.2 g, 3.35 mmol) and (1S)-1-[5-(methoxymethyl)-1,2,4-oxadiazol- 3-yl]-2,2-dimethylpropan-1-amine hydrochloride (1 g, 4.28 mmol) in DMF (30 mL) was added N,N-diisopropylethylamine (2.89 mL, 16.7 mmol) dropwise followed by the addition of HATU (1.91 g, 5.02 mmol). The resulting mixture was stirred at RT for 16 hours. After 16 hours of reaction mixture was diluted with water (100 mL), extracted with EtOAc (3 x 100 mL). Combined organic layer was washed with water (1 x 80 mL) dried over sodium sulfate, filtered through acotton plug and concentrated under reduced pressure to get the crude which was purified by silica gel column chromatography. Fractions were collected by eluting with 3% MeOH in DCM. Appropriate fractions were combined together and concentrated under reduced pressure to afford tert-butyl N-[(1S)-1-{[(1S)-1-[5-(methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2- dimethylpropyl]carbamoyl}-4-(2-nitro-1H-imidazol-1-yl)butyl]carbamate (552 mg, 1.08 mmol, 32% yield, 92% purity) as a off white solid.

[0540] Step-8. Synthesis of tert-butyl N-[(1S)-4-(2-amino-1H-imidazol-1-yl)-1-{[(1S)-1-[5- (methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2-dimethylpropyl]carbamoyl}butyl]carbamate

[0541] To a solution of tert-butyl N-[(1S)-1-{[(1S)-1-[5-(methoxymethyl)-1,2,4-oxadiazol-3- yl]-2,2-dimethylpropyl]carbamoyl}-4-(2-nitro-1H-imidazol-1-yl)butyl]carbamate (600 mg, 1.08 mmol) in ethyl alcohol (30 mL ) and water (4 mL ) was added ammonium chloride (577 mg, 10.8 mmol) and iron (603 mg, 10.8 mmol). The resulting mixture was heated to 80 °C for 3 hours. After 3 hours an aliquot of the reaction mixture was cooled to RT, filtered through a celite bed, washed thoroughly with MeOH (125 mL). Filtrate was concentrated under reduced pressure, co- evaporated with toluene to get the crude tert-butyl N-[(1S)-4-(2-amino-1H-imidazol-1-yl)-1- {[(1S)-1-[5-(methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2-dimethylpropyl]carbamoyl}butyl] carbamate (255 mg, 531 µmol, 49% yield, 25% purity) as an off-white solid.

[0542] Step 9: (2S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-[(1S)-1-[5-(methoxymethyl)- 1,2,4-oxadiazol-3-yl]-2,2-dimethylpropyl]pentanamide hydrochloride

[0543] To a solution of tert-butyl N-[(1S)-4-(2-amino-1H-imidazol-1-yl)-1-{[(1S)-1-[5- (methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2-dimethylpropyl]carbamoyl}butyl]carbamate (1 g, 521 µmol) in DCM (10 mL ) was added trifluoroacetic acid (5 mL, 65.3 mmol) dropwise at 0 °C. The resulting reaction mixture was stirred at RT for 2 hours. After 2 hours reaction mixture was concentrated under reduced pressure to yield the crude which was purified by preparative HPLC (0.1%TFA in water and acetonitrile) to afford product as a TFA salt. For removing TFA salt, compound was re-purified by preparative HPLC (0.1% HCl in water and acetonitrile). Appropriate fraction was lyophilized to afford (2S)-2-amino-5-(2-amino-1H-imidazol-1-yl)-N-[(1S)-1-[5- (methoxymethyl)-1,2,4-oxadiazol-3-yl]-2,2-dimethylpropyl]pentanamide hydrochloride, (A-58). Yield: 39.7mg, 18% yield; Appearance: White hygroscopic solid; 1H-NMR (400 MHz, DMSO- d6): δ 12.12 (s, 1H), 9.03 (d, J = 8.80 Hz, 1H), 8.44 (d, J = 3.20 Hz, 3H), 7.78 (s, 2H), 6.96 (s, 1H), 6.92 (s, 1H), 4.92 (d, J = 8.80 Hz, 1H), 4.77 (s, 2H), 4.16-4.18 (m, 1H), 3.94-3.84 (m, 2H), 3.36(s, 3H), 1.76-1.65 (m, 4H), 1.00 (s, 9H); HPLC purity: 99.4%; LCMS Calculated for C17H29N7O3: 379.23; Observed: 380.2 [M+H]+. Example A14. Synthesis of (S)-1-(2-amino-5-(2-amino-1H-imidazol-1-yl)pentanamido)-N- methylcyclohexane-1-carboxamide (A-59)

[0544] Step 1. Synthesis of methyl 1-aminocyclohexane-1-carboxylate

[0545] To a stirring solution of 1-aminocyclohexane-1-carboxylic acid (1 g, 6.98 mmol) in Methanol (12 ml) was added thionyl chloride (2.54 mL, 34.9 mmol) and a drop of DMF at 0 °C and heated to 65°C for 4h. Reaction monitoring by TLC and LCMS showed complete consumption of starting material and formation of desired product mass. The reaction mixture was then concentrated under reduced pressure to afford the desired intermediate (1.06 g, 6.74 mmol, 88.4% purity, 97.2% yield) as a waxy solid.

[0546] Step 2. Synthesis of methyl (S)-1-(2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H- imidazol-1-yl)pentanamido)cyclohexane-1-carboxylate

[0547] To a stirring solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H- imidazol-1-yl)pentanoic acid, INT-3 (2.00 g, 6.10 mmol) in ethyl acetate (4 mL) and then was added triethylamine hydrochloride (4.19 g, 30.5 mmol). The reaction mixture was cooled to 0 °C and T3P (2.42 g, 7.63 mmol) and methyl 1-aminocyclohexane-1- carboxylate (1.2 g, 7.63 mmol) were added and stirred at room temperature for 16 h. Then the reaction mixture quenched with water (50 mL) and extracted with EtOAc (2 x 150 mL). The combined organic layers were concentrated under reduced pressure and the obtained crude was purified via silica gel column chromatography. Elution of the column using 10-15% EtOAc in petroleum ether and the concentration of column fractions under the reduced pressure afforded the desired intermediate (1.6 g, 3.42 mmol, 44.9% yield, 97.8% purity) as gummy solid.

[0548] Step-3. Synthesis of (S)-1-(2-((tert-butoxycarbonyl)amino)-5-(2-nitro-1H-imidazol-1- yl)pentanamido)cyclohexane-1-carboxylic acid (exp.32)

[0549] To a stirring solution of methyl 1-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro- 1H-imidazol-1-yl)pentanamido]cyclohexane-1- carboxylate (1.4 g, 2.99 mmol) in THF (50 mL) and water (50 mL) was added lithium hydroxide. Mono hydrate (499 mg, 11.9 mmol) and stirred at room temperature for 16 h. The reaction mixture was concentrated under the reduced pressure, cooled with ice-cold water, and then was added 1.5N HCl dropwise till pH~6. Formation of white solid was observed which was then filtered and then dried under vacuum for overnight to afford the desired intermediate (1.29 g, 2.86 mmol, 99.6% purity, 95.5% yield) as a white solid.

[0550] Step 4. Synthesis of tert-butyl (S)-(1-((1-(methylcarbamoyl)cyclohexyl)amino)-5-(2- nitro-1H-imidazol-1-yl)-1-oxopentan-2-yl)carbamate

[0551] To a stirring solution of 1-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-(2-nitro-1H- imidazol-1-yl)pentanamido]cyclohexane-1-carboxylic acid (200 mg, 441 µmol) in ethyl acetate (5 mL) was added triethylamine (222 mg, 2.20 mmol) and cooled to 0 °C. Then T3P (560 mg, 882 µmol) and methanamine, hydrochloride (89.1 mg, 1.32 mmol) was added, and the above reaction mixture was stirred for 16 h at RT. Reaction mixture was diluted with Ethyl acetate (35 mL) and washed with water (2 x 10 mL). The organic layer was dried over anhydrous sodium sulphate, filtered, and concentrated under the reduced pressure to afford the desired intermediate (155 mg, 334 µmol, 91.7% purity, 75.6% yield) as white solid.

[0552] Step 5. Synthesis of tert-butyl (S)-(1-((1-(methylcarbamoyl)cyclohexyl)amino)-5-(2- nitro-1H-imidazol-1-yl)-1-oxopentan-2-yl)carbamate

[0553] To a solution of tert-butyl N-[(1S)-1-{[1-(methylcarbamoyl)cyclohexyl]carbamoyl}-4- (2-nitro-1H-imidazol-1-yl)butyl]carbamate (170 mg, 332 µmol) in methanol (3.0 ml) was added Pd / C (42.3 mg, 39.8 µmol) which was triturated with methanol before adding. The reaction mixture was stirred at RT for 16 h under hydrogen atmosphere. Reaction mixture was filtered through celite bed and was washed with MeOH (3 x 30 mL). The obtained filtrate was concentrated under reduced pressure to get the desired intermediate (162 mg, 372 µmol, 112% yield, 81.4% purity) ) as a pale yellow liquid.

[0554] Step 6. Synthesis of (S)-1-(2-amino-5-(2-amino-1H-imidazol-1-yl)pentanamido)-N- methylcyclohexane-1-carboxamide

[0555] To an ice-cooled solution of tert-butyl N-[(1S)-4-(2-amino-1H-imidazol-1-yl)-1-{[1- (methylcarbamoyl)cyclohexyl]carbamoyl}butyl]carbamate (200 mg, 371 µmol) in DCM ( 2 mL ) was added 4M HCL in Dioxane (41.0 mg, 1.11 mmol) over 5 min. The resulting mixture was warmed to room temperature and stirred at RT for 6 h. Reaction mixture was concentrated under reduced pressure and the obtained crude was purified by prep-HPLC purification to afford (S)-1- (2-amino-5-(2-amino-1H-imidazol-1-yl)pentanamido)-N-methylcyclohexane-1-carboxamide, (A- 59) as a HCl salt (26 mg, 76.8 µmol, 99.5% purity, 20.8% yield). Appearance: White solid. LCMS purity: 99.55%; HPLC purity: 99.72%.1H-NMR (400 MHz, DMSO-d6): δ 12.00 (s, 1H), 8.44 (s, 1H), 8.35 (br s, 3H), 7.74 (s, 2H), 7.61 (d, J = 4.80 Hz, 1H), 7.04 (s, 1H), 6.95 (s, 1H), 4.05 (br s, 1H), 3.94-3.95 (m, 2H), 2.55 (d, J = 4.80 Hz, 3H), 2.17-2.20 (m, 1H), 1.70-1.72 (m, 6H), 1.50- 1.55 (m, 6H), 1.19-0.00 (m, 2H), . LCMS calculated for C16H28N6O2: 336.44; Observed: 337.2 [M+H]+. Example A15. Synthesis of 1-acetyl-3-[(2S)-2-amino-5-(2-amino-1H-imidazol-1- yl)pentanamido]-N-methylpyrrolidine-3-carboxamide; formic acid salt, (A-60)

[0556] Step-1. Synthesis of methyl 1-acetyl-3-((tert-butoxycarbonyl)amino)pyrrolidine-3- carboxylate

[0557] To a solution of methyl 3-{[(tert-butoxy)carbonyl]amino}pyrrolidine-3-carboxylat...

Claims

CLAIMS 1. A compound of formula I: A-B-C I or a pharmaceutically acceptable salt thereof, wherein A is a moiety that binds or associates with p62; B is a linker moiety; and C is a target binding moiety.

2. The compound of claim 1, wherein A is a polypeptide or peptidomimetic moiety that binds or associates with p62.

3. The compound of claims 1 or 2, wherein A is a dipeptide or peptidomimetic moiety that binds or associates with p62.

4. The compound of claim 1, wherein the compound is of formula II-1 or II-2:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatomsselected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, - ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety.

5. The compound of claim 1, wherein the compound is of formula III-1 or III-2:or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatomsselected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa; B is a linker moiety; and C is a target binding moiety.

6. The compound of claims 4 or 5, wherein G1is optionally substituted 5- to 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S.

7. The compound of claims 4 or 5, wherein G1is 5- to 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S, optionally substituted with one or more of –N(Ra)2, or halogen.

8. The compound of claims 4 or 5, wherein G1is 7 to 12-membered bicyclic heteroaryl comprising 1 to 4 heteroatoms selected from N, O, and S, optionally substituted with one or more –N(Ra)2, or halogen.

9. The compound of any one of claims 4-8, wherein the compound is of formula II-1 or III- 1.

10. The compound of claim 9, wherein G1is selected from:

11. The compound of any one of claims 4-8, wherein the compound is of formula II-2 or III- 2.

12. The compound of claim 11, wherein G1is selected from:wherein represents a point of attachment to moiety B, and represents a point of attachment to moiety G2.

13. The compound of any one of claims 4-12, wherein G2is C1-C6aliphatic or C1-C6aliphatic-4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, wherein G2is optionally substituted with one or more Rb.

14. The compound of claim 13, wherein G2is C1-C6aliphatic optionally substituted with one or more Rb.

15. The compound of claim 14, wherein G2is C1-C6aliphatic optionally substituted with one or more –N(Ra)2.

16. The compound of claim 15, wherein G2is C1-C6aliphatic, substituted with –NH2.

17. The compound of claim 16, wherein G2is C1-C6aliphatic, substituted with –NH2, wherein the carbon atom bonded to -NH2is in an S enantiomeric configuration.

18. The compound of claim 13, wherein G2is C1-C6aliphatic-4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, wherein G2is optionally substituted with one or more Rb.

19. The compound of claim 18, wherein G2is –CH2-4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S.

20. The compound of any one of claims 4-12, wherein G2is selected from:whereinrepresents a point of attachment to –C(O)-N(Ra)- in formulae II-1 or II-2, or to A in formulae III-1 or III-2, andrepresents a point of attachment to G1.

21. The compound of any one of claims 4-20, wherein G3is C1-C7aliphatic-C(O)N(Ra)- or 4- to 7-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, wherein G3is optionally substituted with one or more Rc.

22. The compound of claim 21, wherein G3is C1-C7aliphatic-C(O)N(Ra) optionally substituted with halogen or an optionally substituted C1-C6aliphatic.

23. The compound of claim 22, wherein G3is C1-C7alkyl-C(O)N(Ra) optionally substituted with halogen or an optionally substituted C1-C6aliphatic.

24. The compound of claims 9, 10, or 13-20, wherein G3is selected from:, wherein --- represents a point of attachment to moiety B.

25. The compound of any one of claims 11-20, wherein G3is selected from:

26. The compound of claim 24, wherein G3is selected from:wherein --- represents a point ofattachment to moiety B.

27. The compound of claim 25, wherein G3is selected from:

28. The compound of any one of claims 5-27, wherein Ring A is :

29. The compound of claim 4, wherein the compound is of formula IVa:or a pharmaceutically acceptable salt thereof.

30. The compound of claim 4, wherein the compound is of formula IVbor a pharmaceutically acceptable salt thereof.

31. The compound of claim 4, wherein the compound is of formula IVc-1:or a pharmaceutically acceptable salt thereof.

32. The compound of claim 4, wherein the compound is of formula IVd-1:or a pharmaceutically acceptable salt thereof.

33. The compound of claim 4, wherein the compound is of formula IVe-1:or a pharmaceutically acceptable salt thereof.

34. The compound of claim 4, wherein the compound is of formula IVf:or a pharmaceutically acceptable salt thereof.

35. The compound of claim 4, wherein the compound is of formula IVg:or a pharmaceutically acceptable salt thereof.

36. The compound of claim 4, wherein the compound is of formula IVhor a pharmaceutically acceptable salt thereof, wherein ach of W1, W2, W3, and W4is independently selected from N, CH, and CRb,.

37. The compound of claim 4, wherein the compound is of formula IVior a pharmaceutically acceptable salt there.

38. The compound of claim 4, wherein the compound is of formula IVjor a pharmaceutically acceptable salt thereof, wherein each of W5, W6, W7, and W8is independently selected from N and CRd,wherein each Rdis indepdnently selected from H, – N(Ra)2, optionally substituted C1-C6aliphatic, halogen, or moiety B-C wherein one of W1, W2, W3or W4is CRd, and Rdis moiety B-C.

39. The compound of any one of claims 1-38, wherein the linker is an optionally substituted C2-30aliphatic group wherein one or more carbons are optionally and independently replaced by - Cy-, -NRZ-, -N(RZ)C(O)-, -C(O)N(RZ)-, -N(RZ)C(O)O-, -OC(O)N(RZ)-, -N(RZ)C(O)N(RZ) -, - OC(O)O-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -SO-, -SO2-, wherein each -Cy- is independently an optionally substituted 3-12 membered bivalent heterocyclyl ring having 1-3 heteroatoms selected from N, O, and S, an optionally substituted 3-8 membered bivalent heteroaryl ring having 1-4 heteroatoms selected from N, O, and S, an optionally substituted C3-C6cycloalkyl, or an optionally substituted C6-C12aryl, and each RZis independently H or an optionally substituted group selected from C1-C20aliphatic, or C3-C12cycloaliphatic.

40. The compound of any one of claims 1-39, wherein B is a linker moiety selected from Table B.

41. The compound of any one of claims 1-40, wherein C is a is a target binding moiety that binds to ALK, BRD4, p62, Myddosome (IRAK4), Myddosome (MALT1), FGFR, RET, HTT, EML4, Tau, NLRP3 Inflammasome, EGFR / RTK, Androgen Receptor, ACC2, KMO, IAPP, TSPO, STING, cGAS, or USP30.

42. The compound of claim 1, wherein the compound is selected from Table 1.

43. A pharmaceutical composition comprising a compound of any one of claims 1-42, and a pharmaceutically acceptable carrier, filler, or diluent.

44. A method of treating a disease, disorder, or condition in a subject comprising administering a compound of any one of claims 1-42, or the pharmaceutical composition of claim 43.

45. The method of claim 44, wherein the disease, disorder, or condition is selected from NASH, NAFLD, cancer (e.g., cervical cancer, colon cancer, breast cancer, lung cancer, stomach cancer, gastrointestinal cancer, pancreatic cancer, prostate cancer, leukemia, melanoma, lymphoma), Burkitt lymphoma, active B-cell-like diffuse large B-cell lymphomas, diffuse large B-cell lymphomas, primary central nervous system lymphomas, IgM-secreting lymphoplasmacytic lymphoma, Waldenstrom macroglobulinemia, gout, atherosclerosis, Alzheimer’s disease, diabetes (e.g., Type II diabetes), experimental autoimmune encephalitis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, cryopyrin-associated periodic syndromes, Parkinson’s disease, Dementia with Lewy bodies, multiple systems atrophy, neuroaxonal dystrophies, primary age-related tauopathy (PART) dementia, chronic traumatic encephalopathy, Progressive supranuclear palsy (PSP), Corticobasal degeneration (CBD), Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), Lytico-bodig disease (Parkinson-dementia complex of Guam), Ganglioglioma and gangliocytoma, Meningioangiomatosis, Postencephalitic parkinsonism, Subacute sclerosing panencephalitis (SSPE), Lead encephalopathy, Tuberous sclerosis, Pantothenate kinase-associated neurodegeneration, Lipofuscinosis, Spinal and bulbar muscular atrophy (SBMA) / Kennedy’s disease, rheumatoid arthritis, psoriasis, Systemic lupus erythematosus, Aicardia-Goutieressyndrome, ataxia, Familial chilblain lupus, Huntington’s disease, Spinocerebellar ataxia, Familial amylotrophic lateral sclerosis, Frontotemporal dementia (FTLD-TDP), and Amyotrophic lateral sclerosis.

46. A method of inducing degradation of a target in a biological sample, comprising contacting the biological sample with a compound of any one of claims 1-42, or a pharmaceutical composition of claim 43.

47. A compound of any one of claims 1-42, or a pharmaceutical composition of claim 43, for use in medicine.

48. Use of a compound of any one of claims 1-42, or a pharmaceutical composition of claim 43, in the treatment of a disease, disorder, or condition.

49. The use of claim 48, wherein the disease, disorder, or condition is NASH, NAFLD, cancer (e.g., cervical cancer, colon cancer, breast cancer, lung cancer, stomach cancer, gastrointestinal cancer, pancreatic cancer, prostate cancer, leukemia, melanoma, lymphoma), Burkitt lymphoma, active B-cell-like diffuse large B-cell lymphomas, diffuse large B-cell lymphomas, primary central nervous system lymphomas, IgM-secreting lymphoplasmacytic lymphoma, Waldenstrom macroglobulinemia, gout, atherosclerosis, Alzheimer’s disease, diabetes (e.g., Type II diabetes), experimental autoimmune encephalitis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, cryopyrin-associated periodic syndromes, Parkinson’s disease, Dementia with Lewy bodies, multiple systems atrophy, neuroaxonal dystrophies, primary age-related tauopathy (PART) dementia, chronic traumatic encephalopathy, Progressive supranuclear palsy (PSP), Corticobasal degeneration (CBD), Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), Lytico-bodig disease (Parkinson-dementia complex of Guam), Ganglioglioma and gangliocytoma, Meningioangiomatosis, Postencephalitic parkinsonism, Subacute sclerosing panencephalitis (SSPE), Lead encephalopathy, Tuberous sclerosis, Pantothenate kinase-associated neurodegeneration, Lipofuscinosis, Spinal and bulbar muscular atrophy (SBMA) / Kennedy’s disease, rheumatoid arthritis, psoriasis, Systemic lupus erythematosus, Aicardia-Goutieres syndrome, ataxia, Familial chilblain lupus, Huntington’sdisease, Spinocerebellar ataxia, Familial amylotrophic lateral sclerosis, Frontotemporal dementia (FTLD-TDP), and Amyotrophic lateral sclerosis 50. A compound of formula X:or a pharmaceutically acceptable salt thereof, wherein: G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa.

51. A compound of formula XI:or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O, and S; G1is 5- to 12-membered heteroaryl comprising 1 to 6 heteroatoms selected from N, O, and S, 4- to 6-membered heterocycle comprising 1 to 4 heteroatoms selected from N, O, and S, C6-C12aryl, guanidine, –C(O)NH2, or –C(NH)NH2, wherein G1is optionally substituted with one or more Rb; G2is C1-C6aliphatic, -N(Ra)-C1-C6aliphatic, C1-C6aliphatic-N(Ra)-, -O-C1-C6aliphatic, -C(O)-C1-C6aliphatic, C6-C12aryl-C0-C6aliphatic, 2- to 10-membered heteroaliphatic, or (4- to 6-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S)-C1-C6aliphatic, wherein G2is optionally substituted with one or more Rb; G3is C1-C7aliphatic, C1-C7aliphatic-C(O)N(Ra)-C0-C6aliphatic, C3-C12cycloaliphatic, 4- to 9-membered heterocycle comprising 1- to 3- heteroatoms selected from N, O, and S, C1-C7aliphatic-C(O)-4- to 9-membered heterocycle comprising 1 to 3 heteroatoms selected from N, O, and S, or S(O)2, wherein G3is optionally substituted with one or more Rc; each Rais independently selected from H and optionally substituted C1-C6aliphatic; each Rbis independently selected from –N(Ra)2, optionally substituted C1-C6aliphatic, and halogen; each Rcis independently selected from optionally substituted C1-C6aliphatic, optionally substituted C3-C6cycloaliphatic, optionally substituted C6-C12aryl, optionally substituted 4- to 12- membered heteroaryl, optionally substituted 4- to 6-membered heterocycle, -ORa, -C(O)-C1-C6aliphatic, and -C(O)-ORa.

52. The compound of claims 50 or 51, wherein the compound is selected from Table 2.

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