Anti-HER2 immunoconjugates and uses thereof

Abstract

The present invention provides immunoconjugates of Formula I comprising an anti-HER2 antibody linked by conjugation to one or more 8-Het-2-aminobenzazepine derivatives. The present invention also provides 8-Het-2-aminobenzazepine derivative intermediate compositions comprising reactive functional groups. Such intermediate compositions are suitable substrates for forming immunoconjugates via linkers or linking moieties. The present invention further provides methods for treating cancer using the immunoconjugates.

Description

Technical field

[0001] Cross-reference of related applications This application claims the benefit of priority to U.S. Provisional Application No. 63 / 124,367, filed December 11, 2020, which is incorporated by reference in its entirety.

[0002] The invention generally relates to immunoconjugates comprising an anti-HER2 antibody conjugated to one or more 8-Het-2-aminobenzazepine molecules. [Background technology]

[0003] New compositions and methods for the delivery of antibodies and immune adjuvants are needed to reach inaccessible tumors and / or expand treatment options for cancer patients and other subjects. The invention provides such compositions and methods. [Summary of the invention]

[0004] The invention is generally directed to immunoconjugates comprising an anti-HER2 antibody linked by conjugation to one or more 8-Het-2-aminobenzazepine derivatives. The invention is further directed to 8-Het-2-aminobenzazepine derivative intermediate compositions containing reactive functional groups. Such an intermediate composition is a suitable substrate for the formation of an immunoconjugate, in which the antibody, by linker L, has the formula: [ka] 8-Het-2-aminobenzazepine (HxBz) moiety, where Het is selected from heterocyclyldiyl and heteroaryldiyl, and R 1 ,R 2 ,R 3 , and R 4 One of them is connected to L. R 1-4 and X 1-4 Substituents are defined herein.

[0005] The invention is further directed to the use of such immunoconjugates in the treatment of diseases, particularly cancer.

[0006] One aspect of the invention is an immunoconjugate comprising an antibody covalently attached to a linker that is covalently attached to one or more 8-Het-2-aminobenzazepine moieties.

[0007] Another embodiment of the invention is 8-Het-2-aminobenzazepine-linker compounds.

[0008] Another aspect of the invention comprises administering a therapeutically effective amount of an immunoconjugate comprising an anti-HER2 antibody linked by conjugation to one or more 8-Het-2-aminobenzazepine moieties. This is a method for treating cancer.

[0009] Another aspect of the invention is the use of an immunoconjugate comprising an anti-HER2 antibody linked by conjugation to one or more 8-Het-2-aminobenzazepine moieties to treat cancer.

[0010] Another embodiment of the invention is a method of preparing an immunoconjugate by conjugation of one or more 8-Het-2-aminobenzazepine moieties with an anti-HER2 antibody. [Form for carrying out the invention]

[0011] Reference will now be made in detail to specific embodiments of the invention, examples of which are illustrated in the accompanying structures and formulas. While the invention will be described in conjunction with the enumerated embodiments, it will be understood that they are not intended to limit the invention to those embodiments. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents that may fall within the scope of the invention as defined by the claims.

[0012] Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. The invention is not limited to the methods and materials described.

[0013] definition The term "immunoconjugate" or "immunostimulatory antibody conjugate" refers to an antibody construct that is covalently attached to an adjuvant moiety via a linker. The term "adjuvant" refers to a substance that is capable of eliciting an immune response in a subject exposed to the adjuvant.

[0014] "Adjuvant moiety" refers to an adjuvant that is covalently attached to an antibody construct via a linker, eg, as described herein. The adjuvant moiety can elicit an immune response while bound to the antibody construct or after cleavage (eg, enzymatic cleavage) from the antibody construct following administration of the immune complex to a subject.

[0015] "Adjuvant" refers to a substance that is capable of eliciting an immune response in a subject exposed to the adjuvant.

[0016] The terms "Toll-like receptor" and "TLR" refer to any member of a family of highly conserved mammalian proteins that recognize pathogen-associated molecular patterns and function as important signaling elements in innate immunity. . TLR polypeptides share a characteristic structure that includes an extracellular domain with leucine-rich repeats, a transmembrane domain, and an intracellular domain involved in TLR signaling.

[0017] The terms "Toll-like receptor 7" and "TLR7" refer to publicly available TLR7 sequences, e.g., GenBank accession number AAZ99026 for human TLR7 polypeptide, or GenBank accession number AAK62676 for mouse TLR7 polypeptide. Refers to nucleic acids or polypeptides that share at least about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or more sequence identity.

[0018] The terms "Toll-like receptor 8" and "TLR8" refer to publicly available TLR7 sequences, e.g., GenBank accession number AAZ95441 for human TLR8 polypeptide, or GenBank accession number AAK62677 for mouse TLR8 polypeptide. Refers to nucleic acids or polypeptides that share at least about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or more sequence identity.

[0019] A "TLR agonist" is a substance that directly or indirectly binds to TLR (eg, TLR7 and / or TLR8) and induces TLR signal transduction. Any detectable difference in TLR signaling can indicate that the agonist stimulates or activates TLRs. Differences in signal transduction may be associated with, for example, expression of target genes, phosphorylation of signaling components, subcellular localization of downstream elements such as nuclear factor-κB (NF-κB), and specific components (IL-1 receptor-associated It can manifest as changes in the association of kinases (such as IRAK) with other proteins or intracellular structures, or in the biochemical activity of components such as kinases (such as mitogen-activated protein kinases (MAPK)).

[0020] "Antibody" refers to a polypeptide that includes an antigen-binding region (including complementarity determining regions (CDRs)) from an immunoglobulin gene or fragment thereof. The term "antibody" specifically refers to monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibodies that exhibit a desired biological activity. Contains fragments. Exemplary immunoglobulin (antibody) structural units include tetramers. Each tetramer is composed of two identical pairs of polypeptide chains, each pair consisting of one "light" (approximately 25 kDa) and one "heavy" chain (approximately 50 kDa) connected by disulfide bonds. ~70kDa). Each chain is made up of structural domains called immunoglobulin domains. These domains vary in size and function, e.g., variable domains or regions on the light and heavy chains (respectively, V L and V H ), and constant domains or regions on the light and heavy chains (respectively, C L and C H ) are classified into different categories. The N-terminus of each chain defines a variable region of approximately 100-110 or more amino acids called a paratope, which is primarily responsible for antigen recognition, ie, the antigen-binding domain. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which define the immunoglobulin classes IgG, IgM, IgA, IgD, and IgE, respectively. IgG antibodies are large molecules of approximately 150 kDa consisting of four peptide chains. IgG antibodies contain two identical class gamma heavy chains of about 50 kDa and two identical light chains of about 25 kDa, thus a tetrameric quaternary structure. The two heavy chains are linked to each other by disulfide bonds and each to a light chain. The resulting tetramer has two identical halves, which together form a Y-like shape. Each end of the fork contains the same antigen binding domain. There are four IgG subclasses in humans (IgG1, IgG2, IgG3, and IgG4), named in order of their abundance in serum (ie, IgG1 is the most abundant). Typically, the antigen-binding domain of an antibody will be most important in the specificity and affinity of binding to cancer cells.

[0021] "Antibody construct" refers to an antibody or fusion protein that includes (i) an antigen binding domain and (ii) an Fc domain.

[0022] In some embodiments, the binding agent is an antigen-binding antibody "fragment," which is a construct that includes at least the antigen-binding region of an antibody, alone or together with other components that constitute an antigen-binding construct. be. For example, (i)V L ,V H ,C L , and CH 1 (ii) F(ab’), a bivalent fragment consisting of two Fab fragments connected by a disulfide bridge in the hinge region; 2 fragment, (iii) the V of a single arm of the antibody. L and V H (iv) F(ab’) using mild reducing conditions 2 Fab' fragments resulting from breaking the disulfide bridges of the fragments, (v) disulfide-stabilized Fv fragments (dsFv), and (vi) allowing the two domains to be synthesized as a single polypeptide chain. The two domains of the Fv fragment (i.e., V L and V H Many different types of antibody "fragments" are known in the art, including single chains (scFvs), which are monovalent molecules consisting of (scFv).

[0023] The antibody or antibody fragment can be part of a larger construct, eg, a conjugate or fusion construct of the antibody fragment to additional regions. For example, in some embodiments, antibody fragments can be fused to an Fc region as described herein. In other embodiments, the antibody fragment (e.g., Fab or scFv) includes, for example, a transmembrane domain (optionally with an intervening linker or "stalk" (e.g., hinge region)) and any cell-to-cell signaling. By fusion to a domain, it can be part of a chimeric antigen receptor or a chimeric T cell receptor.

[0024] "Epitope" means any antigenic or epitopic determinant of an antigen to which an antigen-binding domain binds (ie, in a paratope of the antigen-binding domain). Antigenic determinants usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three-dimensional structural characteristics and specific charge characteristics.

[0025] The term "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. There are three major classes of Fc receptors: (1) FcγR, which binds IgG, (2) FcαR, which binds IgA, and (3) FcεR, which binds IgE. The FcγR family includes several members such as FcγI (CD64), FcγRIIA (CD32A), FcγRIIB (CD32B), FcγRIIIA (CD16A), and FcγRIIIB (CD16B). Fcγ receptors have different affinities for IgG and also have different affinities for IgG subclasses (eg, IgG1, IgG2, IgG3, and IgG4).

[0026] Nucleic acid or amino acid sequence "identity" as referred to herein can be determined by comparing the nucleic acid or amino acid sequence of interest to a reference nucleic acid or amino acid sequence. Percent identity measures the number of nucleotides or amino acid residues that are the same (i.e., identical) between an optimally aligned sequence of interest and a reference sequence over the length of the longest sequence (i.e., the sequence of interest or the length of the reference array (whichever is longer). Sequence alignments and percent identity calculations can be performed using available software programs. Examples of such programs include CLUSTAL-W, T-Coffee, ALIGN (for alignment of nucleic acid and amino acid sequences), BLAST programs (e.g. BLAST2.1, BL2SEQ, BLASTp, BLASTn, etc.), and FASTA programs (e.g. , FASTA3x, FASTM, and SSEARCH) (for sequence alignments and sequence similarity searches). Sequence alignment algorithms are also used, e.g., Altschul et al., J. Molecular Biol., 215(3):403-410(1990), Beigert et al., Proc. Natl. Acad. Sci. USA, 106(10). :3770-3775(2009), Durbin et al., eds., Biological Sequence Analysis: Probalistic Models of Proteins and Nucleic Acids, Cambridge University Press, Cambridge, UK (2009), Soding, Bioinformatics, 21(7):951- 960 (2005), Altschul et al., Nucleic Acids Res., 25(17):3389-3402 (1997), and Gusfield, Algorithms on Strings, Trees and Sequences, Cambridge University Press, Cambridge UK (1997). sea ​​bream. The percent (%) identity of a sequence can also be calculated as, for example, 100x[(identical positions) / min(TG A , T.G. B )] can be calculated as TG A and T.G. B is T.G. A and T.G. B is the sum of the number of residues and internal gap positions in peptide sequences A and B in the alignment that minimizes . See, eg, Russell et al., J. Mol Biol., 244:332-350 (1994).

[0027] The binding agent includes Ig heavy chain and light chain variable region polypeptides that together form an antigen binding site. Each heavy and light chain variable region is a polypeptide that includes three complementarity determining regions (CDR1, CDR2, and CDR3) connected by framework regions. The binding agent can be any of the various types of binding agents known in the art, including Ig heavy and light chains. For example, a binding agent can be an antibody, an antigen-binding antibody "fragment," or a T cell receptor.

[0028] "Biosimilar" means an approved antibody construct that has similar activity properties to a HER2-targeting antibody, such as, for example, trastuzumab (HERCEPTIN™, Genentech, Inc.) or pertuzumab (PERJETA, Genentech, Inc.). refers to

[0029] "Biobetter" refers to an approved antibody construct that is an improvement on a previously approved antibody construct, such as labetuzumab. A biobetter can have one or more modifications to a previously approved antibody construct (eg, an altered glycan profile, or a unique epitope). Biobetters are recombinant protein drugs in the same class as existing biopharmaceuticals, but they are not identical and are better than the original. Biobetter is not just a new drug, nor is it a generic version of a drug. Biosimilars and biobetters are both variants of a biological drug, with the former being a close copy of the original drug and the latter improving in terms of efficacy, safety, and tolerability or dosing regimen. has been done.

[0030] "Amino acid" refers to any monomer unit that can be incorporated into a peptide, polypeptide, or protein. Amino acids include naturally occurring α-amino acids and their stereoisomers, as well as non-naturally occurring (non-naturally occurring) amino acids and their stereoisomers. A "stereoisomer" of a given amino acid refers to isomers that have the same molecular formula and intramolecular bonding, but different three-dimensional arrangements of bonds and atoms (eg, l-amino acids and corresponding d-amino acids). Amino acids can be glycosylated (eg, N-linked glycans, O-linked glycans, phosphoglycans, C-linked glycans, or glypication) or deglycosylated. Amino acids may be referred to herein by either the commonly known three letter symbol or the one letter symbol recommended by the IUPAC-IUB Biochemical Nomenclature Commission.

[0031] Naturally occurring amino acids are those encoded by the genetic code, as well as amino acids that are subsequently modified, such as hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine. Naturally occurring alpha amino acids include alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), Arginine (Arg), Lysine (Lys), Leucine (Leu), Methionine (Met), Asparagine (Asn), Proline (Pro), Glutamine (Gln), Serine (Ser), Threonine (Thr), Valine (Val), Examples include, but are not limited to, tryptophan (Trp), tyrosine (Tyr), and combinations thereof. Naturally occurring stereoisomers of α-amino acids include, but are not limited to, D-alanine (D-Ala), D-cysteine ​​(D-Cys), D-aspartic acid (D-Asp), and D-glutamic acid ( D-Glu), D-phenylalanine (D-Phe), D-histidine (D-His), D-isoleucine (D-Ile), D-arginine (D-Arg), D-lysine (D-Lys), D-leucine (D-Leu), D-methionine (D-Met), D-asparagine (D-Asn), D-proline (D-Pro), D-glutamine (D-Gln), D-serine (D -Ser), D-threonine (D-Thr), D-valine (D-Val), D-tryptophan (D-Trp), D-tyrosine (D-Tyr), and combinations thereof.

[0032] Naturally occurring amino acids include those formed in proteins by post-translational modifications such as citrulline (Cit).

[0033] Non-natural (non-naturally occurring) amino acids include, but are not limited to, amino acid analogs, amino acid mimetics, synthetic amino acids, N-substituted glycines in either the L or D configuration that function in a manner similar to naturally occurring amino acids. , and N-methyl amino acids. For example, an "amino acid analog" has the same basic chemical structure as a naturally occurring amino acid (i.e., hydrogen, carboxyl group, carbon attached to an amino group) but with modified side chain groups or It can be an unnatural amino acid with a peptide backbone, such as homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. "Amino acid mimetic" refers to a compound that has a structure that differs from the general chemical structure of an amino acid, but functions in a manner similar to a naturally occurring amino acid.

[0034] "Linker" refers to a functional group that covalently joins two or more moieties in a compound or material. For example, a linking moiety can serve to covalently attach an adjuvant moiety to an antibody construct in an immunoconjugate.

[0035] "Linking moiety" refers to a functional group that covalently bonds two or more moieties in a compound or material. For example, a linking moiety can serve to covalently attach an adjuvant moiety to an antibody in an immune complex. Useful linkages for linking linking moieties to proteins and other materials include, but are not limited to, amides, amines, esters, carbamates, ureas, thioethers, thiocarbamates, thiocarbonates, and thioureas.

[0036] "Divalent" refers to a chemical moiety that contains two points of attachment for linking two functional groups; a multivalent linking moiety can have additional points of attachment for linking additional functional groups. can. Divalent radicals may be designated with the suffix "diyl". For example, divalent linking moieties include divalent polymeric moieties such as divalent poly(ethylene glycol), divalent cycloalkyl, divalent heterocycloalkyl, divalent aryl, and divalent heteroaryl groups. "Divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group" means a cycloalkyl, heterocycloalkyl, heterocycloalkyl, or heterocycloalkyl group having two points of attachment for covalently linking two moieties within a molecule or material. Refers to an aryl or heteroaryl group. A cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be substituted or unsubstituted. A cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be substituted with one or more groups selected from halo, hydroxy, amino, alkylamino, amido, acyl, nitro, cyano, and alkoxy.

[0037] wavy line [ka] represents the point of attachment of the specified chemical moiety. It will be understood that when two squiggly lines are present in a designated chemical moiety, the chemical moiety may be used to be read on both sides, i.e., left to right or right to left.

[0038] "Alkyl" refers to a straight or branched saturated aliphatic radical having the indicated number of carbon atoms. Alkyl can contain any number of carbons, for example 1-12. Examples of alkyl groups include methyl (Me, -CH 3 ), ethyl (Et, -CH 2 CH 3 ), 1-propyl (n-Pr, n-propyl, -CH 2 CH 2 CH 3 ), 2-propyl(i-Pr, i-propyl, -CH(CH 3 ) 2 ), 1-butyl (n-Bu, n-butyl, -CH 2 CH 2 CH 2 CH 3 ), 2-methyl-1-propyl (i-Bu, i-butyl, -CH 2 CH(CH 3 ) 2 ), 2-butyl(s-Bu, s-butyl, -CH(CH 3 )CH 2 CH 3 ), 2-methyl-2-propyl(t-Bu, t-butyl, -C(CH 3 ) 3 ), 1-pentyl (n-pentyl, -CH 2 CH 2 CH 2 CH 2 CH 3 ), 2-pentyl(-CH(CH 3 )CH 2 CH 2 CH 3 ), 3-pentyl(-CH(CH 2 CH 3 ) 2 ), 2-methyl-2-butyl(-C(CH 3 ) 2 CH 2 CH 3 ), 3-methyl-2-butyl(-CH(CH 3 )CH(CH 3 ) 2 ), 3-methyl-1-butyl(-CH 2 CH 2 CH(CH 3 ) 2 ), 2-methyl-1-butyl(-CH 2 CH(CH 3 )CH 2 CH 3 ), 1-hexyl(-CH 2 CH 2 CH 2 CH 2 CH 2 CH 3 ), 2-hexyl(-CH(CH 3 )CH 2 CH 2 CH 2 CH 3 ), 3-hexyl(-CH(CH 2 CH 3 )(CH 2 CH 2 CH 3 )), 2-methyl-2-pentyl(-C(CH 3 ) 2 CH 2 CH 2 CH 3 ), 3-methyl-2-pentyl(-CH(CH 3 )CH(CH 3 )CH 2 CH 3 ), 4-methyl-2-pentyl(-CH(CH 3 )CH 2 CH(CH 3 ) 2 ), 3-methyl-3-pentyl(-C(CH 3 )(CH 2 CH 3 ) 2 ), 2-methyl-3-pentyl(-CH(CH 2 CH 3 )CH(CH 3 ) 2 ), 2,3-dimethyl-2-butyl(-C(CH 3 ) 2 CH(CH 3 ) 2 ), 3,3-dimethyl-2-butyl(-CH(CH 3 )C(CH 3 ) 3 , 1-heptyl, 1-octyl and the like, but are not limited to these. Alkyl groups may be substituted or unsubstituted. A "substituted alkyl" group may be substituted with one or more groups selected from halo, hydroxy, amino, oxo (=O), alkylamino, amido, acyl, nitro, cyano, and alkoxy.

[0039] The term "alkyldiyl" refers to a divalent alkyl radical. Examples of alkyldiyl groups include methylene (-CH 2 -), ethylene (-CH 2 CH2 -), propylene (-CH 2 CH 2 CH 2 -), etc., but are not limited to these. Alkyldiyl groups may also be referred to as "alkylene" groups.

[0040] "Alkenyl" refers to a straight or branched, unsaturated aliphatic radical having the number of carbon atoms indicated and at least one carbon-carbon double bond, sp2. Alkenyl can contain from 2 to about 12 or more carbon atoms. Alkenyl groups are radicals with "cis" and "trans" orientations, or alternatively, "E" and "Z" orientations. Examples include ethylenyl or vinyl (-CH=CH 2 ), allyl(-CH 2 CH=CH 2 )=, butenyl, pentenyl, and isomers thereof. Alkenyl groups may be substituted or unsubstituted. A "substituted alkenyl" group may be substituted with one or more groups selected from halo, hydroxy, amino, oxo (=O), alkylamino, amido, acyl, nitro, cyano, and alkoxy.

[0041] The term "alkenylene" or "alkenyldiyl" refers to a straight or branched divalent hydrocarbon radical. Examples include ethylenylene or vinylene (-CH=CH-), allyl (-CH 2 Examples include, but are not limited to, CH=CH-).

[0042] "Alkynyl" refers to a straight or branched unsaturated aliphatic radical having the number of carbon atoms indicated and at least one carbon-carbon triple bond, sp. Alkynyl can contain from 2 to about 12 or more carbon atoms. For example, C 2 -C 6 Alkynyl is ethynyl (-C≡CH), propynyl (propargyl, -CH 2 C≡CH), butynyl, pentynyl, hexynyl, and isomers thereof. Alkynyl groups may be substituted or unsubstituted. A "substituted alkynyl" group may be substituted with one or more groups selected from halo, hydroxy, amino, oxo (=O), alkylamino, amido, acyl, nitro, cyano, and alkoxy.

[0043] The term "alkynylene" or "alkynyldiyl" refers to a divalent alkynyl radical.

[0044] The terms "carbocycle," "carbocyclyl," "carbocycle," and "cycloalkyl" refer to saturated or partially unsaturated, monocyclic rings containing from 3 to 12 ring atoms, or the number of atoms indicated. , refers to a fused bicyclic, or bridged polycyclic ring assembly. Saturated monocyclic carbocyclic rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Saturated bicyclic and polycyclic carbocyclic rings include, for example, norbornane, [2.2.2]bicyclooctane, decahydronaphthalene, and adamantane. Carbocyclic groups can also be partially unsaturated, having one or more double or triple bonds within the ring. Representative carbocyclic groups that are partially unsaturated include cyclobutene, cyclopentene, cyclohexene, cyclohexadiene (1,3- and 1,4-isomers), cycloheptene, cycloheptadiene, cyclooctadiene (1 , 3-, 1,4- and 1,5-isomers), norbornene, and norbornadiene.

[0045] The term "cycloalkyldiyl" refers to a divalent cycloalkyl radical.

[0046] "Aryl" is a group of 6 to 20 carbon atoms (C 6 ~C 20 ) refers to a monovalent aromatic hydrocarbon radical. Aryl groups may be monocyclic, fused to form bicyclic or tricyclic groups, or linked by bonds to form biaryl groups. Representative aryl groups include phenyl, naphthyl, and biphenyl. Other aryl groups include benzyl having a methylene linking group. Some aryl groups have 6 to 12 ring members, such as phenyl, naphthyl, or biphenyl. Other aryl groups have 6 to 10 ring members such as phenyl or naphthyl.

[0047] The term "arylene" or "aryldiyl" refers to a group of 6 to 20 carbon atoms (C 6 ~C 20 ) means a divalent aromatic hydrocarbon radical. Some aryldiyl groups are represented as "Ar" in exemplary structures. Aryldiyl includes bicyclic radicals containing saturated, partially unsaturated rings, or aromatic rings fused to aromatic carbocyclic rings. Typical aryldiyl groups include radicals derived from benzene (phenyldiyl), substituted benzene, naphthalene, anthracene, biphenylene, indenylene, indanilene, 1,2-dihydronaphthalene, 1,2,3,4-tetrahydronaphthyl, etc. These include, but are not limited to: Aryldiyl groups are also referred to as "arylene" and are optionally substituted with one or more substituents as described herein.

[0048] The terms "heterocycle," "heterocyclyl," and "heterocyclic ring" are used interchangeably herein and refer to a heteroatom in which at least one ring atom is selected from nitrogen, oxygen, phosphorous, and sulfur. a saturated or partially unsaturated (i.e., having one or more double and / or triple bonds within the ring) carbocyclic ring of from 3 to about 20 ring atoms, with the remaining ring atoms being C Refers to a radical in which one or more ring atoms are optionally substituted independently with one or more substituents described below. Heterocycles are monocyclic rings having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 4 heteroatoms selected from N, O, P, and S), or 7 to 10 bicyclo with ring members (4 to 9 carbon atoms and 1 to 6 heteroatoms selected from N, O, P, and S), e.g., bicyclo[4,5], [5, 5], [5,6], or [6,6] system. Heterocyclic rings are described in Paquette, Leo A.; “Principles of Modern Heterocyclic Chemistry” (W.A. Benjamin, New York, 1968), especially chapters 1, 3, 4, 6, 7, and 9, “The Chemistry of Heterocyclic Compounds, A series of Monographs” (John Wiley&Sons, New York, 1950 to present), especially volumes 13, 14, 16, 19, and 28, and J. Am. Chem. Soc. (1960) 82:5566. "Heterocyclyl" also includes radicals in which a heterocyclic radical is fused with a saturated, partially unsaturated ring, or an aromatic carbocyclic or heterocyclic ring. Examples of heterocycles are morpholin-4-yl, piperidin-1-yl, piperazinyl, piperazin-4-yl-2-one, piperazin-4-yl-3-one, pyrrolidin-1-yl, thiomorpholin- 4-yl, S-dioxothiomorpholin-4-yl, azocan-1-yl, azetidin-1-yl, octahydropyrido[1,2-a]pyrazin-2-yl, [1,4]diazepane -1-yl, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, homopiperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl , thiepanil, oxazepinyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofura Nyl, pyrazolidinylimidazolinyl, imidazolidinyl, 3-azabishico[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 3H-indolylquinolidinyl, and N- These include, but are not limited to, pyridyl urea. Spiroheterocyclyl moieties are also included within this definition. Examples of spiroheterocyclyl moieties include azaspiro[2.5]octanyl and azaspiro[2.4]heptanyl. Examples of heterocyclic groups in which two ring atoms are substituted with oxo (=O) moieties are pyrimidinonyl and 1,1-dioxo-thiomorpholinyl. The heterocyclic groups herein are optionally substituted independently with one or more substituents as described herein.

[0049] The term "heterocyclyldiyl" refers to a dicyclic group of from 3 to about 20 ring atoms in which at least one ring atom is a heteroatom selected from nitrogen, oxygen, phosphorus, and sulfur, and the remaining ring atoms are C. refers to a valent, saturated, or partially unsaturated (i.e., having one or more double and / or triple bonds in the ring) carbocyclic radical, in which one or more ring atoms are optionally independently substituted with one or more substituents, as indicated. Examples of 5- and 6-membered heterocyclyldiyl include morpholinyldiyl, piperidinyldiyl, piperazinyldiyl, pyrrolidinyldiyl, dioxanyldiyl, thiomorpholyldiyl, and S-dioxothiomol. An example is horyldiyl.

[0050] The term "heteroaryl" refers to a 5-, 6-, or 7-membered monovalent aromatic radical containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur. Contains a fused ring system of 20 atoms (at least one is aromatic). Examples of heteroaryl groups are pyridinyl (including, for example, 2-hydroxypyridinyl), imidazolyl, imidazopyridinyl, pyrimidinyl (including, for example, 4-hydroxypyrimidinyl), pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, Thienyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, sinolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl , thiadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. Heteroaryl groups are optionally substituted independently with one or more substituents as described herein.

[0051] The term "heteroaryldiyl" refers to a 5-, 6-, or 7-membered divalent aromatic radical containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur. Contains a fused ring system of 5 to 20 atoms (at least one is aromatic). Examples of 5- and 6-membered heteroaryldiyl include pyridyldiyl, imidazolyldiyl, pyrimidyldiyl, pyrazolyldiyl, triazolyldiyl, pyrazinyldiyl, tetrazolyldiyl, furyldiyl, thienyldiyl, isoxazolyldiyl, thiazolyldiyl, oxadiazole. Included are lyldiyl, oxazolyldiyl, isothiazolyldiyl, and pyrrolyldiyl.

[0052] A heterocycle or heteroaryl group may be carbon (carbon-bonded) or nitrogen (nitrogen-bonded) bonded, where such is possible. By way of example and not limitation, a carbon-bonded heterocycle or heteroaryl can be pyridine at position 2, 3, 4, 5, or 6, pyridazine at position 3, 4, 5, or 6, pyrimidine at position 2, 4, 5, or 6, positions 2, 3, 5, or 6 of pyrazine; positions 2, 3, 4, or 5 of furan, tetrahydrofuran, thiofuran, thiophene, pyrrole, or tetrahydropyrrole; positions 2, 4 of oxazole, imidazole, or thiazole; or 5, isoxazole, pyrazole, or isothiazole in position 3, 4, or 5; aziridine in position 2 or 3; azetidine in position 2, 3, or 4; quinoline in position 2, 3, 4, 5, 6 , 7, or 8, or 3, 4, 5, 6, 7, or 8 of isoquinoline.

[0053] By way of example and not limitation, nitrogen-bonded heterocycles or heteroaryls include aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2- Bonded at position 1 of pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, 1H-indazole, position 2 of isoindole or isoindoline, position 4 of morpholine, and position 9 of carbazole or β-carboline.

[0054] The terms "halo" and "halogen" by themselves or as part of another substituent refer to a fluorine, chlorine, bromine, or iodine atom.

[0055] The term "carbonyl", by itself or as part of another substituent, means C(=O) or -C(=O)-, i.e., any other group within the moiety that is double bonded to oxygen and has a carbonyl. refers to a carbon atom bonded to two groups.

[0056] As used herein, the phrase "quaternary ammonium salt" refers to an alkyl substituent (e.g., C 1 ~C 4 Refers to a tertiary amine that has been quaternized with alkyl).

[0057] The terms "treat," "treatment," and "treating" refer to alleviation, amelioration, alleviation of symptoms, or making a symptom, injury, pathology, or disease more tolerable by a patient, or reducing the rate of progression of symptoms. an injury, pathology, disease (e.g. cancer), or condition, including any objective or subjective parameter, such as reducing, reducing the frequency or duration of a symptom or disease, or in some circumstances preventing the onset of a symptom. Refers to any sign of success in treating or ameliorating (e.g., cognitive impairment). Treatment or amelioration of symptoms may be based on any objective or subjective parameter, including, for example, the results of a physical exam.

[0058] The terms "cancer," "neoplasm," and "tumor" refer to the autonomous, uncontrolled growth of cells in which they exhibit an abnormal growth phenotype characterized by a significant loss of control over cell proliferation. Used herein to refer to cells that exhibit Cells of interest for detection, analysis and / or treatment in the context of the invention include cancer cells (e.g. cancer cells from individuals with cancer), malignant cancer cells, pre-metastatic cancer cells, Includes metastatic cancer cells and non-metastatic cancer cells. Cancers of almost all tissues are known. The phrase "cancer burden" refers to the amount of cancer cells or cancer volume in a subject. Therefore, reducing cancer burden refers to reducing the number of cancer cells or cancer cell volume in a subject. As used herein, the term "cancer cell" is a cancer cell (e.g., from any cancer that an individual may be treated with, e.g., that may be isolated from an individual who has cancer) ), or any cell derived from a cancer cell, e.g., a clone of a cancer cell. For example, cancer cells may be from an established cancer cell line, may be primary cells isolated from an individual with cancer, or may be primary cells isolated from an individual with cancer. For example, it may be a progeny cell from a cell. In some embodiments, the term may also refer to a portion of a cancer cell, a subcellular portion of a cancer cell, a cell membrane portion, or a cell lysate. Many types of cancer are known to those skilled in the art, including solid tumors such as carcinoma, sarcoma, glioblastoma, melanoma, lymphoma, and myeloma, and circulating cancers such as leukemia.

[0059] As used herein, the term "cancer" refers to solid tumor cancers (e.g., skin, lung, prostate, breast, stomach, bladder, colon, ovary, pancreas, kidney, liver, glioblastoma, marrow). blastoma, leiomyosarcoma, head and neck squamous cell carcinoma, melanoma, and neuroendocrine), and liquid cancer (e.g., blood cancer), carcinoma, soft tissue tumor, sarcoma, metamorphoma, melanoma, leukemia, lymphoma, and microscopic Includes any form of cancer that includes residual disease and includes, but is not limited to, brain cancer, including both primary and metastatic tumors.

[0060] Cancer "pathology" includes all phenomena that impair patient well-being. These include abnormal or uncontrolled cell proliferation, metastasis, interference with the normal function of neighboring cells, release of cytokines or other secreted products at abnormal levels, and suppression or exacerbation of inflammatory or immunological responses. including, but not limited to, neoplasms, premalignant tumors, malignant tumors, and invasion of surrounding or distant tissues or organs, such as lymph nodes.

[0061] As used herein, the phrases "cancer recurrence" and "tumor recurrence" and grammatical variants thereof refer to the further growth of neoplastic or cancerous cells after diagnosis of cancer. . In particular, recurrence may occur when further cancerous cell growth occurs in the cancerous tissue. Similarly, "tumor spread" occurs when cells of a tumor spread to local or distant tissues and organs, and thus includes tumor metastasis. "Tumor invasion" occurs when tumor growth spreads locally and impairs the function of the involved tissues by compacting, destroying, or preventing normal organ function.

[0062] As used herein, the term "metastasis" refers to the growth of a cancerous tumor in an organ or body part that is not directly connected to the organ of the original cancerous tumor. Metastasis will be understood to include micrometastasis, which is the presence of undetectable amounts of cancerous cells in an organ or body part that is not directly connected to the organ of the original cancerous tumor. Metastasis can also be defined as several steps in the process, such as detachment of cancer cells from the original tumor site and migration and / or invasion of cancer cells to other parts of the body.

[0063] The phrases "effective amount" and "therapeutically effective amount" refer to the dose or amount of a substance, such as an immune complex, to which it is administered that produces a therapeutic effect. The exact dose depends on the purpose of treatment and can be ascertained by one skilled in the art using known techniques (e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992), Lloyd, The Art, Science and Technology of Pharmaceutical Compounding(1999), Pickar,Dosage Calculations(1999), Goodman&Gilman's The Pharmacological Basis of Therapeutics,11 th Edition (McGraw-Hill, 2006), and Remington: The Science and Practice of Pharmacy, 22 ndEdition, (Pharmaceutical Press, London, 2012). ) In the case of cancer, a therapeutically effective amount of an immune complex reduces the number of cancer cells, reduces tumor size, inhibits (i.e. slows to some extent, and preferably slows) the invasion of cancer cells into peripheral organs. may inhibit (i.e., slow, preferably halt) tumor metastasis to some extent, inhibit tumor growth to some extent, and / or alleviate to some extent one or more of the symptoms associated with cancer. . To the extent that the immune complex can prevent the growth and / or death of existing cancer cells, it may be cytostatic and / or cytotoxic. In the case of cancer treatment, efficacy can be measured, for example, by assessing time to disease progression (TTP) and / or determining response rate (RR).

[0064] "Recipient," "individual," "subject," "host," and "patient" are used interchangeably and refer to any mammalian subject (e.g., human) for whom diagnosis, treatment, or therapy is desired. Point. "Mammal" means any animal classified as a mammal, including humans, domesticated animals, and farm animals, as well as zoo animals such as dogs, horses, cats, cows, sheep, goats, pigs, camels, or sporting animals; Refers to pet animals. In certain embodiments, the mammal is a human.

[0065] The term "synergistic adjuvant" or "synergistic combination" in the context of the present invention refers to receptor agonists, cytokines, and adjuvants that, in combination, elicit a synergistic effect on immunity compared to the administration of either alone. Contains a combination of two immunomodulatory agents such as polypeptides. In particular, the immunoconjugates disclosed herein include a synergistic combination of the claimed adjuvant and antibody construct. These synergistic combinations upon administration induce a greater effect on immunity compared to, for example, when the antibody construct or adjuvant is administered in the absence of the other moiety. Additionally, there was a reduction (as measured by the total number of antibody constructs or the total number of adjuvants administered as part of an immune complex) compared to when either the antibody construct or the adjuvant were administered alone. amount of immune complex may be administered.

[0066] As used herein, the term "administering" refers to parenteral, intravenous, intraperitoneal, intramuscular, intratumoral, intralesional, intranasal, or subcutaneous administration, oral administration, as a suppository, etc. refers to administration, topical contact, intrathecal administration, or implantation of a sustained release device, such as a mini-osmotic pump, into the subject.

[0067] When used herein to modify numerical values, the terms "about" and "approximately" indicate a close range around the numerical value. Therefore, when "X" is a value, "about X" or "approximately References to “about X” or “approximately Be specific. Thus, "about X" and "approximately X" are intended to teach and provide written descriptive support for a claim limitation of, for example, "0.98X."

[0068] HER2 antibody The immunoconjugate of the invention includes an antibody construct that includes an antigen binding domain that specifically recognizes and binds HER2.

[0069] In certain embodiments, the immunoconjugate of the invention comprises an anti-HER2 antibody. In one embodiment of the invention, the anti-HER2 antibodies of the immunoconjugates of the invention are humanized anti-HER2 antibodies, such as those described in Table 3 of US5821337, which is specifically and expressly incorporated herein by reference. Includes huMAb4D5-1, huMAb4D5-2, huMAb4D5-3, huMAb4D5-4, huMAb4D5-5, huMAb4D5-6, huMAb4D5-7, and huMAb4D5-8. These antibodies contain human framework regions with the complementarity determining regions of a murine antibody (4D5) that binds HER2. The humanized antibody huMAb4D5-8, also referred to as trastuzumab, is commercially available under the trade name HERCEPTIN™ (Genentech, Inc.).

[0070] Trastuzumab (CAS180288-69-1, HERCEPTIN®, huMAb4D5-8, rhuMAb HER2, Genentech) inhibits the extracellular domain of HER2 (US5677171, US5821337, US6054297, US6165464, US6339142, US6407213, US6639055, US6719971, US6800738, US7074404, Coussens et al(1985)Science230:1132-9, Slamon et al(1989)Science244:707-12, Slamon et al(2001)New Engl.J.Med.3 44 IgG1 kappa, a recombinant DNA-derived monoclonal antibody, is a humanized version of the murine anti-HER2 antibody (4D5) that selectively binds with high affinity to 4D5.

[0071] In one embodiment of the invention, the antibody construct or antigen binding domain comprises the CDR regions of trastuzumab. In one embodiment of the invention, the anti-HER2 antibody further comprises a framework region of trastuzumab. In one embodiment of the invention, the anti-HER2 antibody further comprises one or both variable regions of trastuzumab.

[0072] In another embodiment of the invention, the anti-HER2 antibody of the immunoconjugate of the invention comprises a humanized anti-HER2 antibody, such as humanized 2C4, as described in US7862817. An exemplary humanized 2C4 antibody is pertuzumab (CAS Reg. No. 380610-27-5), PERJETA™ (Genentech, Inc.). Pertuzumab is a HER dimerization inhibitor (HDI) in which HER2 forms active heterodimers or homodimers with other HER receptors (such as EGFR / HER1, HER2, HER3, and HER4) It functions to inhibit ability. For example, Harari and Yarden, Oncogene19:6102-14(2000); Yarden and Sliwkowski.Nat Rev Mol Cell Biol2:127-37(2001), Sliwkowski Nat Struct Biol10:158-9(2003), Cho et al.Nature421: 756-60 (2003), and Malik et al. Pro Am Soc Cancer Res 44:176-7 (2003). PERJETA™ is approved for the treatment of breast cancer.

[0073] In one embodiment of the invention, the antibody construct or antigen binding domain comprises the CDR regions of pertuzumab. In one embodiment of the invention, the anti-HER2 antibody further comprises a pertuzumab framework region. In one embodiment of the invention, the anti-HER2 antibody further comprises one or both variable regions of pertuzumab.

[0074] The immunoconjugates of the invention include antibodies that target, bind, or recognize HER2. Within the scope of embodiments of the invention are functional variants of the antibody constructs or antigen binding domains described herein. As used herein, the term "functional variant" refers to an antibody construct that has an antigen binding domain that has substantial or significant sequence identity or similarity to a parent antibody construct or antigen binding domain; A functional variant retains the biological activity of the antibody construct or antigen binding domain of which it is a variant. A functional variant, e.g., as described herein, retains the ability to recognize target cells expressing HER2 to a similar extent, to the same extent, or to a higher extent than the parent antibody construct or antigen binding domain. (parent antibody construct or antigen binding domain).

[0075] With respect to an antibody construct or antigen binding domain, a functional variant is, for example, at least about 30%, about 50%, about 75%, about 80%, about 85%, about 90% different in amino acid sequence from the antibody construct or antigen binding domain. %, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more.

[0076] A functional variant can include, for example, a parent antibody construct or antigen binding domain amino acid sequence with at least one conservative amino acid substitution. Alternatively or additionally, a functional variant can include an amino acid sequence of a parent antibody construct or antigen binding domain that has at least one non-conservative amino acid substitution. In this case, it is preferred that the non-conservative amino acid substitutions do not interfere with or inhibit the biological activity of the functional variant. Non-conservative amino acid substitutions can enhance the biological activity of a functional variant such that the biological activity of the functional variant is increased compared to the parent antibody construct or antigen binding domain.

[0077] Antibodies comprising the immunoconjugates of the invention include Fc engineered variants. In some embodiments, mutations in the Fc region that result in regulated binding to one or more Fc receptors include the following mutations: SD(S239D), SDIE(S239D / I332E), SE(S267E), SELF( S267E / L328F), SDIE(S239D / I332E), SDIEAL(S239D / I332E / A330L), GA(G236A), ALIE(A330L / I332E), GASDALIE(G236A / S239D / A330L / I332E), V9(G237D / P238D / P271G / A330R), and one or more of V11 (G237D / P238D / H268D / P271G / A330R), and / or the following amino acids: 1 in E345R, E233, G237, P238, H268, P271, L328, and A330 can contain more than one mutation. Additional Fc region modifications to modulate Fc receptor binding are described, for example, in U.S. Patent Application No. 2016 / 0145350 and U.S. Patent Nos. 7,416,726 and 5,624,821, which are incorporated by reference in their entirety Incorporated herein.

[0078] Antibodies containing the immunoconjugates of the invention include carbohydrate chain variants such as afucosylation. In some embodiments, the Fc region of the binding agent is modified to have an altered glycosylation pattern of the Fc region compared to the native, unmodified Fc region.

[0079] Amino acid substitutions in antibody constructs or antigen binding domains of the invention are preferably conservative amino acid substitutions. Conservative amino acid substitutions are known in the art, in which one amino acid with specific physical and / or chemical properties is replaced with another amino acid with the same or similar chemical or physical properties. Contains amino acid substitutions. For example, conservative amino acid substitutions include an acidic / loaded polar amino acid (e.g. Asp or Glu) substituted with another acidic / loaded polar amino acid, a nonpolar side substituted with another amino acid with a nonpolar side chain. Amino acids with chains (e.g. Ala, Gly, Val, Ile, Leu, Met, Phe, Pro, Trp, Cys, Val, etc.), basic / positive polarity substituted with another basic / positive polarity amino acid Amino acids (e.g. Lys, His, Arg, etc.), uncharged amino acids with a polar side chain substituted by another uncharged amino acid with a polar side chain (e.g. Asn, Gln, Ser, Thr, Tyr, etc.), beta Beta Amino acids with branched side chains substituted with another amino acid with a branched side chain (e.g., Ile, Thr, and Val), Amino acids with an aromatic side chain substituted with another amino acid with an aromatic side chain (eg, His, Phe, Trp, and Tyr).

[0080] An antibody construct or antigen binding domain can consist essentially of the specified amino acid sequence(s) described herein, such that other components, e.g. does not substantially alter the biological activity of the construct or antigen-binding domain functional variant.

[0081] In some embodiments, the antibody in the immunoconjugate contains a modified Fc region, and the modification modulates the binding of the Fc region to one or more Fc receptors.

[0082] In some embodiments, the antibody in the immune complex (e.g., an antibody conjugated to at least two adjuvant moieties) has one or more Fc receptors compared to a native antibody that lacks mutations in the Fc region. (e.g., increased binding or decreased binding) The Fc region contains one or more modifications (eg, amino acid insertions, deletions, and / or substitutions) that result in the Fc region. In some embodiments, the antibody in the immune complex has one or more modifications within the Fc region (e.g., amino acid insertions, deletions, and / or substitution). In some embodiments, the antibody in the immune complex reduces binding of the antibody to FcγRIIB compared to a native antibody lacking mutations in the Fc region, while FcγRI(CD64), FcγRIIA(CD32A) , and / or one or more modifications (e.g., amino acid insertions, deletions, and / or substitutions) within the Fc region of the antibody that maintain or have increased binding to FcRγIIIA (CD16a). contains. In some embodiments, the antibody in the immunoconjugate contains one or more modifications within the Fc region that increase binding of the Fc region of the antibody to FcγRIIB.

[0083] In some embodiments, modulated binding is provided by mutations within the Fc region of the antibody compared to the native Fc region of the antibody. The mutation can be in the CH2 domain, CH3 domain, or a combination thereof. "Native Fc region" is synonymous with "wild-type Fc region" and is the same as the amino acid sequence of an Fc region found in nature or the amino acid sequence of an Fc region found in a naturally occurring antibody (e.g., cetuximab). Contains identical amino acid sequences. Native sequence human Fc regions include native sequence human IgG1 Fc regions, native sequence human IgG2 Fc regions, native sequence human IgG3 Fc regions, and native sequence human IgG4 Fc regions, as well as naturally occurring variants thereof. Natural sequence Fc includes various allotypes of Fc (Jefferis et al., (2009) mAbs, 1(4):332-338).

[0084] In some embodiments, mutations in the Fc region that result in modulated binding to one or more Fc receptors include the following mutations: SD(S239D), SDIE(S239D / I332E), SE(S267E), SELF (S267E / L328F), SDIE(S239D / I332E), SDIEAL(S239D / I332E / A330L), GA(G236A), ALIE(A330L / I332E), GASDALIE(G236A / S239D / A330L / I332E), V9(G237D / P238D / P271G / A330R), and one or more of V11 (G237D / P238D / H268D / P271G / A330R), and / or one of the following amino acids: E233, G237, P238, H268, P271, L328, and A330 The above mutations can be included. Additional Fc region modifications to modulate Fc receptor binding are described, for example, in US2016 / 0145350 and US7416726 and US5624821, which are incorporated herein by reference in their entirety.

[0085] In some embodiments, the Fc region of the antibody of the immunoconjugate is modified to have an altered glycosylation pattern of the Fc region compared to the native, unmodified Fc region.

[0086] Human immunoglobulins are glycosylated at residue Asn297 within the Cγ2 domain of each heavy chain. This N-linked oligosaccharide is composed of the core heptasaccharide, N-acetylglucosamine 4 mannose 3 (GlcNAc4Man3). Removal of heptasaccharides by endoglycosidase or PNGase F is known to result in conformational changes in the antibody Fc region, which significantly reduces antibody binding affinity for FcγR activation and reduces effector function. Can be done. Core heptasaccharides are often decorated with galactose, bisected GlcNAc, fucose, or sialic acid, which differentially affects Fc binding to activating and inhibitory FcγRs. Furthermore, α2,6-sialylation enhances anti-inflammatory activity in vivo, while defucosylation leads to improved FcγRIIIa binding and a 10-fold increase in antibody-dependent cytotoxicity and phagocytosis. This has been proven. Thus, specific glycosylation patterns can be used to control inflammatory effector functions.

[0087] In some embodiments, the modification to alter the glycosylation pattern is a mutation. For example, substitution with Asn297. In some embodiments, Asn297 is mutated to glutamine (N297Q). Methods for controlling immune responses with antibodies that modulate FcγR-regulated signaling are described, for example, in U.S. Pat. are incorporated herein by reference in their entirety.

[0088] In some embodiments, the antibodies of the immunoconjugate are modified to contain engineered Fab regions with non-naturally occurring glycosylation patterns. For example, hybridomas can be genetically engineered to secrete afucosylated mAbs, deallylated mAbs, or deglycosylated Fc with specific mutations that allow for increased FcRγIIIa binding and effector function. In some embodiments, the antibodies of the immunoconjugate are engineered to be afucosylated.

[0089] In some embodiments, the entire Fc region of the antibody within the immune complex is exchanged with a different Fc region, such that the Fab region of the antibody is conjugated to the non-native Fc region. In some embodiments, Fc-modified antibodies with non-natural Fc domains also include one or more amino acid modifications, such as the S228P mutation within the IgG4Fc, that modulate the stability of the described Fc domain. In some embodiments, Fc-modified antibodies with non-natural Fc domains also include one or more amino acid modifications described herein that modulate Fc binding to FcR.

[0090] In some embodiments, a modification that modulates the binding of an Fc region to an FcR does not alter the binding of an antibody's Fab region to its antigen when compared to a native, unmodified antibody. In other embodiments, modifications that modulate the binding of the Fc region to FcR also increase the binding of the Fab region of the antibody to its antigen when compared to a native, unmodified antibody.

[0091] In some embodiments, the antibody in the immunoconjugate is glycosylated.

[0092] In some embodiments, the antibodies in the immunoconjugate contain engineered cysteines at sites where they are available for conjugation but do not disrupt immunoglobulin folding and assembly, or antigen binding and effector function. Cysteine-engineered antibodies that provide site-specific conjugation of adjuvants, labels, or drug moieties through substitution (Junutula, et al., 2008b Nature Biotech., 26(8):925-932, Dornan et al. (2009) Blood 114(13):2721-2729, US7521541, US7723485, US2012 / 0121615, WO2009 / 052249). A "cysteine ​​engineered antibody" or "cysteine ​​engineered antibody variant" is an antibody in which one or more residues of the antibody are replaced with a cysteine ​​residue. The cysteine ​​engineered antibody can be conjugated to the 8-Het-2-aminobenzazepine adjuvant moiety as an 8-Het-2-aminobenzazepine-linker compound with uniform stoichiometry (e.g., a single For antibodies with one engineered cysteine ​​site, up to two 8-Het-2-aminobenzazepine moieties per antibody).

[0093] In some embodiments, the cysteine ​​engineered antibodies used to prepare the immunoconjugates of Table 3 have a cysteine ​​residue introduced at the 149-lysine site of the light chain (LC K149C). In other embodiments, the cysteine ​​engineered antibody has a cysteine ​​residue introduced at the 118-alanine site (EU numbering) (HC A118C) of the heavy chain. This site is alternatively numbered 121 by sequential numbering or 114 by Kabat numbering. In other embodiments, cysteine ​​engineered antibodies have cysteine ​​residues introduced in the light chain of G64C or R142C according to Kabat numbering, or in the heavy chain of D101C, V184C, or T205C according to Kabat numbering.

[0094] 8-HET-2-aminobenzazepine adjuvant compound The immunoconjugate of the invention comprises an 8-Het-2-aminobenzazepine adjuvant moiety. An adjuvant moiety described herein is a compound that elicits an immune response (ie, an immunostimulant). Generally, the adjuvant moieties described herein are TLR agonists. TLRs are type I transmembrane proteins involved in the initiation of innate immune responses in vertebrates. TLRs recognize a variety of pathogen-associated molecular patterns from bacteria, viruses, and fungi and serve as the first line of defense against invading pathogens. TLRs elicit overlapping but distinct biological responses due to differences in cellular expression and the signaling pathways they initiate. When engaged (e.g., by natural stimuli or synthetic TLR agonists), TLRs activate nuclear factor-κB (NF-κB) and IL-1 through the adapter protein myeloid differentiation primary response gene 88 (MyD88). Initiates a signaling cascade that leads to the recruitment of receptor-associated kinase (IRAK). Phosphorylation of IRAK then leads to recruitment of TNF-receptor-associated factor 6 (TRAF6), which results in phosphorylation of the NF-κB inhibitor I-κB. As a result, NF-κB enters the cell nucleus and initiates transcription of genes whose promoters contain NF-κB binding sites, such as cytokines. Additional regulatory modes for TLR signaling include TIR domain-containing adapter-induced interferon-β (TRIF)-dependent induction of TNF receptor-associated factor 6 (TRAF6) and activation of the MyD88-independent pathway through TRIF and TRAF3. , leading to phosphorylation of interferon response factor 3 (IRF3). Similarly, the MyD88-dependent pathway also activates several IRF family members, including IRF5 and IRF7, while the TRIF-dependent pathway also activates the NF-κB pathway.

[0095] Typically, the adjuvant moieties described herein are TLR7 and / or TLR8 agonists. Both TLR7 and TLR8 are expressed on monocytes and dendritic cells. In humans, TLR7 is also expressed on plasmacytoid dendritic cells (pDCs) and B cells. TLR8 is primarily expressed in cells of myeloid origin, namely monocytes, granulocytes, and myeloid dendritic cells. TLR7 and TLR8 can detect the presence of "foreign" single-stranded RNA within cells as a means of responding to viral invasion. Treatment of TLR8-expressing cells with TLR8 agonists can result in the production of high levels of IL-12, IFN-γ, IL-1, TNF-α, IL-6, and other inflammatory cytokines. Similarly, stimulation of TLR7-expressing cells such as pDCs with TLR7 agonists can result in the production of high levels of IFN-α and other inflammatory cytokines. TLR7 / TLR8 engagement and resultant cytokine production can activate dendritic cells and other antigen-presenting cells and drive diverse innate and acquired immune response mechanisms leading to tumor destruction.

[0096] Exemplary 8-Het-2-aminobenzazepine compounds (Hx) of the invention are shown in Table 1. Each compound was synthesized, purified, characterized by mass spectrometry and shown to have the indicated mass. Additional experimental procedures can be found in the Examples. Activity against HEK293NFKB reporter cells expressing human TLR7 or human TLR8 was measured according to Example 202. The 8-Het-2-aminobenzazepine compounds of Table 1 demonstrate surprising and unexpected properties of TLR8 agonist selectivity that may predict useful therapeutic activity for treating cancer and other disorders. . [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11]

[0097] 8-HET-2-aminobenzazepine-linker compound The immunoconjugate of the invention is prepared by conjugation of an anti-HER2 antibody and an 8-Het-2-aminobenzazepine-linker compound, HxBzL. 8-Het-2-aminobenzazepine-linker compounds include an 8-Het-2-aminobenzazepine (Hx) moiety covalently attached to a linker unit. The linker unit contains functional groups and subunits that affect stability, permeability, solubility, and other pharmacokinetic, safety, and efficacy properties of the immunoconjugate. The linker unit includes a reactive functional group that reacts with, or conjugates to, a reactive functional group of the antibody. For example, a nucleophilic group such as a lysine side chain amino of an antibody reacts with an electrophilic reactive functional group of an Hx linker compound to form an immunoconjugate. Also, for example, cysteine ​​thiol of an antibody reacts with the maleimide or bromoacetamide group of the Hx-linker compound to form an immune complex.

[0098] Suitable electrophilic reactive functional groups for Hx linker compounds include N-hydroxysuccinimidyl (NHS) ester and N-hydroxysulfosuccinimidyl (sulfo-NHS) ester (amine-reactive), carbodiimide (amine-reactive), and carboxyl reactivity), hydroxymethylphosphine (amine reactivity), maleimide (thiol reactivity), halogenated acetamides (thiol reactivity) such as N-iodoacetamide, aryl azides (primary amine reactivity), fluorinated aryl azides (reactivity due to carbon hydrogen (C-H) insertion), pentafluorophenyl (PFP) ester (amine reactivity), terafluorophenyl (TFP) ester (amine reactivity), imido ester (amine reactivity), isocyanate (hydroxyl Examples include, but are not limited to, vinyl sulfones (thiol-, amine-, and hyphoroxyl-reactive), pyridyl disulfides (thiol-reactive), and benzophenone derivatives (reactive via C-H bond insertion). Additional reagents include Hermanson, Bioconjugate Techniques 2 nd Edition, Academic Press, 2008, but is not limited to these.

[0099] The invention provides solutions to the limitations and challenges to the design, preparation, and use of immunoconjugates. Some linkers can be unstable in the bloodstream, thereby releasing an unacceptable amount of adjuvant / drug before internalization within target cells (Khot, A. et al (2015) Bioanalysis 7(13):1633-1648). Other linkers may provide stability in the bloodstream, but intracellular release efficacy may be negatively affected. Linkers that provide the desired intracellular release typically have low stability in the bloodstream. Stated another way, blood flow stability and intracellular release are typically inversely related. Additionally, standard conjugation processes depend on the amount of adjuvant / drug moiety loaded onto the antibody, i.e. drug loading, the amount of aggregates formed in the conjugation reaction, and the yield of final purified conjugate that can be obtained. are interrelated. For example, aggregate formation generally correlates positively with the number of equivalents of adjuvant / drug moieties and derivatives thereof conjugated to the antibody. Under high drug loading, the formed aggregates must be removed for therapeutic use. As a result, drug loading-mediated aggregate formation can reduce immune complex yield and make process scale-up difficult.

[0100] An exemplary embodiment has the formula II: [ka] 8-Het-2-aminobenzazepine-linker compound, wherein Het is selected from heterocyclyldiyl and heteroaryldiyl; R 1 ,R 2 ,R 3 , and R 4 But independently, H, C 1 -C 12 Alkyl, C 2 -C 6 alkenyl, C 2 -C 6 Alkynyl, C 3 -C 12 Carbocyclyl, C 6 -C 20 Aryl, C 2 -C 9 heterocyclyl, and C 1 -C 20 selected from the group consisting of heteroaryl, where alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heterocyclyl, and heteroaryl are independently and optionally -(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 1 -C 12 alkyldiyl)-OR 5 , -(C 3 -C 12 carbocyclyl), -(C 3 -C 12 carbocyclyl)-*, -(C 3 -C 12 carbocyclyl)-(C 1 -C 12 alkyldiyl)-NR 5 -*, -(C 3 -C 12 carbocyclyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 3 -C 12 carbocyclyl)-NR 5 -C(=NR 5 )NR 5 -*, -(C 6 -C 20 aryl), -(C 6 -C 20 aryl)-*, -(C 6 -C 20 aryldiyl)-N(R 5 )-*, -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-(C 2 -C 20 heterocyclyldiyl)-*, -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-NR 5 -C(=NR 5a )N(R 5 )-*, -(C 2 -C 20 heterocyclyl), -(C 2 -C 20 heterocyclyl)-*, -(C 2 -C 9 heterocyclyl)-(C 1 -C 12 alkyldiyl)-NR 5 -*, -(C 2 -C 9 heterocyclyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 2 -C 9 heterocyclyl)-C(=O)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 2 -C 9 heterocyclyl)-NR 5 -C(=NR 5a )NR 5 -*, -(C 2 -C 9 heterocyclyl)-NR 5 -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 2 -C 9 heterocyclyl)-(C 6 -C 20 aryldiyl)-*, -(C 1 -C 20 heteroaryl), -(C 1 -C 20 heteroaryl)-*, -(C 1 -C 20 heteroaryl)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 1 -C 20 heteroaryl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 1 -C 20 heteroaryl)-NR 5 -C(=NR 5a )N(R 5 )-*, -(C 1 -C 20 heteroaryl)-N(R 5 )C(=O)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -C(=O)-*, -C(=O)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -C(=O)-(C 2 -C 20 heterocyclyldiyl)-*, -C(=O)N(R 5 ) 2 , -C(=O)N(R 5 )-*, -C(=O)N(R 5 )-(C 1 -C 12 alkyldiyl)-N(R 5 )C(=O)R 5 , -C(=O)N(R 5 )-(C 1 -C 12 alkyldiyl)-N(R 5 )C(=O)N(R 5 ) 2 , -C(=O)NR 5 -(C 1 -C12 alkyldiyl)-N(R 5 )CO 2 R 5 , -C(=O)NR 5 -(C 1 -C 12 alkyldiyl)-N(R 5 )C(=NR 5a )N(R 5 ) 2 , -C(=O)NR 5 -(C 1 -C 12 alkyldiyl)-NR 5 C(=NR 5a )R 5 , -C(=O)NR 5 -(C 1 -C 8 alkyldiyl)-NR 5 (C 2 -C 5 heteroaryl), -C(=O)NR 5 -(C 1 -C 20 heteroaryldiyl)-N(R 5 )-*, -C(=O)NR 5 -(C 1 -C 20 heteroaryldiyl)-*, -C(=O)NR 5 -(C 1 -C 20 heteroaryldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -C(=O)NR 5 -(C 1 -C 20 heteroaryldiyl)-(C 2 -C 20 heterocyclyldiyl)-C(=O)NR 5 -(C 1 -C 12 alkyldiyl)-NR 5 -*, -N(R 5 ) 2 , -N(R 5 )-*, -N(R 5 )C(=O)R 5 , -N(R 5 )C(=O)-*, -N(R 5 )C(=O)N(R 5 ) 2 , -N(R 5 )C(=O)N(R 5 )-*, -N(R 5 )CO 2 R 5 , -NR 5 C(=NR 5a )N(R 5 ) 2 , -NR 5 C(=NR 5a )N(R 5 )-*, -NR 5 C(=NR 5a )R 5 , -N(R 5 )C(=O)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -N(R 5 )-(C 2 -C 5 heteroaryl), -N(R 5 )-S(=O) 2 -(C 1 -C 12 alkyl), -O-(C 1 -C 12 alkyl), -O-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -O-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -O-C(=O)N(R 5 ) 2 , -O-C(=O)N(R 5 )-*, -S(=O) 2 -(C 2 -C 20 heterocyclyldiyl)-*, -S(=O) 2 -(C 2 -C 20 heterocyclyldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -S(=O) 2 -(C 2 -C 20 heterocyclyldiyl)-(C 1 -C 12 alkyldiyl)-NR 5 -*,as well as -S(=O) 2 -(C 2 -C 20 heterocyclyldiyl)-(C 1 -C 12 substituted with one or more groups selected from alkyldiyl)-OH, or R 2 and R 3 together form a 5- or 6-membered heterocyclyl ring, X 1 ,X 2 ,X 3 , and X 4 but independently, the combination, C(=O), C(=O)N(R 5 ), O, N(R 5 ), S, S(O) 2 , and S(O) 2 N(R 5 ), R 5 But independently, H, C 6 -C 20 Aryl, C 3 -C 12 Carbocyclyl, C 6 -C 20 Aryljiyl, C 1 -C 12 alkyl, and C 1 -C 12 selected from the group consisting of alkyldiyl or two R 5 the groups together form a 5- or 6-membered heterocyclyl ring, R 5a However, C 6 -C 20 Aryl and C 1 -C 20 selected from the group consisting of heteroaryl, In the formula, the asterisk * indicates the binding site of L, and R 1 ,R 2 ,R 3 , and R 4 One of them is bonded to L, L is Q-C(=O)-PEG-, Q-C(=O)-PEG-C(=O)N(R 6 )-(C 1 -C 12 alkyldiyl)-C(=O)-Gluc-, Q-C(=O)-PEG-O-, Q-C(=O)-PEG-O-C(=O)-, Q-C(=O)-PEG-C(=O)-, Q-C(=O)-PEG-C(=O)-PEP-, Q-C(=O)-PEG-N(R 6 )-, Q-C(=O)-PEG-N(R 6 )-C(=O)-, Q-C(=O)-PEG-N(R 6 )-PEG-C(=O)-PEP-, Q-C(=O)-PEG-N + (R 6 ) 2 -PEG-C(=O)-PEP-, Q-C(=O)-PEG-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-, Q-C(=O)-PEG-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)N(R 6 )C(=O)-(C 2 -C 5 monoheterocyclyldiyl)-, Q-C(=O)-PEG-SS-(C 1 -C 12 alkyldiyl)-OC(=O)-, Q-C(=O)-PEG-SS-(C 1 -C 12 alkyldiyl)-C(=O)-, Q-C(=O)-(C 1 -C 12 alkyldiyl)-C(=O)-PEP-, Q-C(=O)-(C 1 -C 12 alkyldiyl)-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-, Q-C(=O)-(C1 -C 12 alkyldiyl)-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-N(R 5 )-C(=O), Q-C(=O)-(C 1 -C 12 alkyldiyl)-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-N(R 6 )C(=O)-(C 2 -C 5 monoheterocyclyldiyl)-, Q-(CH 2 ) m -C(=O)N(R 6 )-PEG-, Q-(CH 2 ) m -C(=O)N(R 6 )-PEG-C(=O)N(R 6 )-(C 1 -C 12 alkyldiyl)-C(=O)-Gluc-, Q-(CH 2 ) m -C(=O)N(R 6 )-PEG-O-, Q-(CH 2 ) m -C(=O)N(R 6 )-PEG-O-C(=O)-, Q-(CH 2 ) m -C(=O)N(R 6 )-PEG-C(=O)-, Q-(CH 2 ) m -C(=O)N(R 6 )-PEG-N(R 5 )-, Q-(CH 2 ) m -C(=O)N(R 6 )-PEG-N(R 5 )-C(=O)-, Q-(CH 2 ) m -C(=O)N(R 6 )-PEG-C(=O)-PEP-, Q-(CH 2 ) m -C(=O)N(R 6 )-PEG-SS-(C 1 -C 12 alkyldiyl)-OC(=O)-, Q-(CH 2 ) m -C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-, Q-(CH 2 ) m -C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)N(R 6 )C(=O)-, and Q-(CH 2 ) m -C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)N(R 6 )C(=O)-(C 2 -C 5 a linker selected from the group consisting of monoheterocyclyldiyl)-, R 6 but independently H or C 1 -C 6 is alkyl, PEG has the formula:-(CH 2 CH 2 O) n -(CH 2 ) m -, m is an integer from 1 to 5, n is an integer from 2 to 50, Gluc has the formula: [ka] has PEP is the formula: [ka] , where AA is independently selected from natural or non-natural amino acid side chains, or one or more of AA and the adjacent nitrogen atom form a 5-membered proline amino acid. form, the wavy line indicates the bonding point, Cyc is optionally F, Cl, NO 2 , -OH, -OCH 3 , and C substituted with one or more groups selected from glucuronic acid having the structure: 6 -C 20 Aryldiyl and C 1 -C 20 selected from heteroaryldiyl, [ka] R 7 But -CH(R 8 )O-, -CH 2 -,-CH 2 N(R 8 )-, and -CH(R 8 )O-C(=O)-, where R 8 But, H, C 1 -C 6 Alkyl, C(=O)-C 1 -C 6 Alkyl, and -C(=O)N(R 9 ) 2 selected from, where R 9 But independently, H, C 1 -C 12 alkyl, and -(CH 2 CH 2 O) n -(CH 2 ) m -OH, where m is an integer from 1 to 5, n is an integer from 2 to 50, or two R 9 the groups together form a 5- or 6-membered heterocyclyl ring, y is an integer from 2 to 12, z is 0 or 1, Q is F, Cl, NO 2 , and S.O. 3 - selected from the group consisting of N-hydroxysuccinimidyl, N-hydroxysulfosuccinimidyl, maleimide, and phenoxy substituted with one or more groups independently selected from Alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl are independently and optionally F, Cl, Br, I, -CN, -CH 3 ,-CH 2 CH 3 ,-CH=CH 2 , -C≡CH, -C≡CCH 3 ,-CH 2 CH 2 CH 3 , -CH(CH 3 ) 2 ,-CH 2 CH(CH 3 ) 2 ,-CH 2 OH, -CH 2 OCH 3 ,-CH 2 CH 2 OH, -C(CH 3 ) 2 OH, -CH(OH)CH(CH 3 ) 2 , -C(CH 3 ) 2 CH 2 OH, -CH 2 CH 2 S.O. 2 CH 3 ,-CH 2 OP(O)(OH) 2 ,-CH 2 F, -CHF 2 ,-CF 3 ,-CH 2 C.F. 3 ,-CH 2 CHF 2 , -CH(CH 3 )CN, -C(CH 3 ) 2 CN, -CH 2 CN, -CH 2 N.H. 2 ,-CH 2 NHSO 2 CH 3 ,-CH 2 NHCH 3 ,-CH 2 N(CH 3 ) 2 ,-CO 2 H,-COCH 3 ,-CO 2 CH 3 ,-CO 2 C(CH 3 ) 3 , -COCH(OH)CH 3 ,-CONH 2 ,-CONHCH 3 ,-CON(CH 3 ) 2 , -C(CH 3 ) 2 CONH 2 ,-NH 2 , -NHCH 3 , -N(CH 3 ) 2 ,-NHCOCH 3 , -N(CH 3)COCH 3 ,-NHS(O) 2 CH 3 , -N(CH 3 )C(CH 3 ) 2 CONH 2 , -N(CH 3 )CH 2 CH 2 S(O) 2 CH 3 , -NHC(=NH)H, -NHC(=NH)CH 3 , -NHC(=NH)NH 2 ,-NHC(=O)NH 2 ,-NO 2 , =O, -OH, -OCH 3 ,-OCH 2 CH 3 ,-OCH 2 CH 2 OCH 3 ,-OCH 2 CH 2 OH, -OCH 2 CH 2 N(CH 3 ) 2 , -O(CH 2 CH 2 O) n -(CH 2 ) m C.O. 2 H, -O(CH 2 CH 2 O) n H, -OCH 2 F,-OCHF 2 ,-OCF 3 , -OP(O)(OH) 2 , -S(O) 2 N(CH 3 ) 2 ,-SCH 3 , -S(O) 2 CH 3 , and -S(O) 3 Substituted with one or more groups independently selected from H.

[0101] Exemplary embodiments of the 8-Het-2-aminobenzazepine-linker compounds of Formula II provide that Q is: [ka] including being selected from.

[0102] Exemplary embodiments of the 8-Het-2-aminobenzazepine-linker compounds of Formula II include that Q is phenoxy substituted with one or more F.

[0103] Exemplary embodiments of the 8-Het-2-aminobenzazepine-linker compounds of Formula II include that Q is 2,3,5,6-tetrafluorophenoxy.

[0104] Exemplary embodiments of 8-Het-2-aminobenzazepine-linker (HxBzL) compounds are selected from Table 2a and Table 2b. Each compound was synthesized, purified, characterized by mass spectrometry and shown to have the indicated mass. Additional experimental procedures can be found in the Examples. The 8-Het-2-aminobenzazepine-linker compounds of Tables 2a and 2b exhibit surprising and unexpected TLR8 agonist selectivity that may predict useful therapeutic activity for treating cancer and other disorders. Demonstrate characteristics that do not. The 8-Het-2-aminobenzazepine-linker intermediates of Tables 2a and 2b, Formula II compounds, are used conjugated with antibodies by the method of Example 201 to form immunoconjugates of Tables 3a and 3b. form. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9] [Table 2-10] [Table 2-11] [Table 2-12] [Table 2-13] [Table 2-14] [Table 2-15] [Table 2-16] [Table 2-17] [Table 2-18] [Table 2-19] [Table 2-20] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9] [Table 3-10] [Table 3-11] [Table 3-12] [Table 3-13] [Table 3-14] [Table 3-15] [Table 3-16] [Table 3-17] [Table 3-18] [Table 3-19] [Table 3-20] [Table 3-21] [Table 3-22] [Table 3-23] [Table 3-24] [Table 3-25] [Table 3-26] [Table 3-27] [Table 3-28] [Table 3-29] [Table 3-30]

[0105] immune complex An exemplary embodiment of an immunoconjugate comprises an anti-HER2 antibody covalently attached to one or more 8-Het-2-aminobenzazepine (Hx) moieties by a linker and has the formula I: Ab-[L-Hx] p I has During the ceremony, the Ab is an antibody construct having an antigen binding domain that binds to HER2; p is an integer from 1 to 8, Hx is the formula: [ka] 8-Het-2-aminobenzazepine moiety with Het selected from heterocyclyldiyl and heteroaryldiyl, R 1 ,R 2 ,R 3 , and R 4 But independently, H, C 1 -C 12 Alkyl, C 2 -C 6 alkenyl, C 2 -C 6 Alkynyl, C 3 -C 12 Carbocyclyl, C 6 -C 20 Aryl, C 2 -C 9 heterocyclyl, and C 1 -C 20 selected from the group consisting of heteroaryl, where alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heterocyclyl, and heteroaryl are independently and optionally -(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 1 -C 12 alkyldiyl)-OR 5 , -(C 3 -C 12 carbocyclyl), -(C 3 -C 12 carbocyclyl)-*, -(C 3 -C 12 carbocyclyl)-(C 1 -C 12 alkyldiyl)-NR 5 -*, -(C 3 -C 12 carbocyclyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 3 -C 12 carbocyclyl)-NR 5 -C(=NR 5 )NR 5 -*, -(C 6 -C 20 aryl), -(C 6 -C 20 aryl)-*, -(C 6 -C 20 aryldiyl)-N(R 5 )-*, -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-(C 2 -C 20 heterocyclyldiyl)-*, -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-NR 5 -C(=NR 5a )N(R 5 )-*, -(C 2 -C 20 heterocyclyl), -(C 2 -C 20 heterocyclyl)-*, -(C 2 -C 9 heterocyclyl)-(C 1 -C 12 alkyldiyl)-NR 5 -*, -(C 2 -C 9 heterocyclyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 2 -C 9 heterocyclyl)-C(=O)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 2 -C 9 heterocyclyl)-NR 5 -C(=NR 5a )NR 5 -*, -(C 2 -C 9 heterocyclyl)-NR 5 -(C 6 -C 20 aryldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 2 -C 9 heterocyclyl)-(C 6 -C 20 aryldiyl)-*, -(C 1 -C 20 heteroaryl), -(C 1 -C 20 heteroaryl)-*, -(C 1 -C 20 heteroaryl)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -(C 1 -C 20 heteroaryl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -(C 1 -C 20 heteroaryl)-NR 5 -C(=NR 5a )N(R 5 )-*, -(C 1 -C 20 heteroaryl)-N(R 5 )C(=O)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -C(=O)-*, -C(=O)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -C(=O)-(C 2 -C 20 heterocyclyldiyl)-*, -C(=O)N(R 5 ) 2 , -C(=O)N(R 5 )-*, -C(=O)N(R 5 )-(C 1 -C 12 alkyldiyl)-N(R 5 )C(=O)R 5 , -C(=O)N(R 5 )-(C 1 -C 12 alkyldiyl)-N(R 5 )C(=O)N(R 5 ) 2 , -C(=O)NR 5 -(C 1 -C 12 alkyldiyl)-N(R 5 )CO 2 R 5 , -C(=O)NR 5 -(C 1 -C 12 alkyldiyl)-N(R 5 )C(=NR 5a )N(R 5 ) 2 , -C(=O)NR 5 -(C 1 -C12 alkyldiyl)-NR 5 C(=NR 5a )R 5 , -C(=O)NR 5 -(C 1 -C 8 alkyldiyl)-NR 5 (C 2 -C 5 heteroaryl), -C(=O)NR 5 -(C 1 -C 20 heteroaryldiyl)-N(R 5 )-*, -C(=O)NR 5 -(C 1 -C 20 heteroaryldiyl)-*, -C(=O)NR 5 -(C 1 -C 20 heteroaryldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -C(=O)NR 5 -(C 1 -C 20 heteroaryldiyl)-(C 2 -C 20 heterocyclyldiyl)-C(=O)NR 5 -(C 1 -C 12 alkyldiyl)-NR 5 -*, -N(R 5 ) 2 , -N(R 5 )-*, -N(R 5 )C(=O)R 5 , -N(R 5 )C(=O)-*, -N(R 5 )C(=O)N(R 5 ) 2 , -N(R 5 )C(=O)N(R 5 )-*, -N(R 5 )CO 2 R 5 , -NR 5 C(=NR 5a )N(R 5 ) 2 , -NR 5 C(=NR 5a )N(R 5 )-*, -NR 5 C(=NR 5a )R 5 , -N(R 5 )C(=O)-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -N(R 5 )-(C 2 -C 5 heteroaryl), -N(R 5 )-S(=O) 2 -(C 1 -C 12 alkyl), -O-(C 1 -C 12 alkyl), -O-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -O-(C 1 -C 12 alkyldiyl)-N(R 5 )-*, -O-C(=O)N(R 5 ) 2 , -O-C(=O)N(R 5 )-*, -S(=O) 2 -(C 2 -C 20 heterocyclyldiyl)-*, -S(=O) 2 -(C 2 -C 20 heterocyclyldiyl)-(C 1 -C 12 alkyldiyl)-N(R 5 ) 2 , -S(=O) 2 -(C 2 -C 20 heterocyclyldiyl)-(C 1 -C 12 alkyldiyl)-NR 5 -*,as well as -S(=O) 2 -(C 2 -C 20 heterocyclyldiyl)-(C 1 -C 12 substituted with one or more groups selected from alkyldiyl)-OH, or R 2 and R 3 together form a 5- or 6-membered heterocyclyl ring, X 1 ,X 2 ,X 3 , and X 4 but independently, the combination, C(=O), C(=O)N(R 5 ), O, N(R 5 ), S, S(O) 2 , and S(O) 2 N(R 5 ), R 5 But independently, H, C 6 -C 20 Aryl, C 3 -C 12 Carbocyclyl, C 6 -C 20 Aryljiyl, C 1 -C 12 alkyl, and C 1 -C 12 selected from the group consisting of alkyldiyl or two R 5 the groups together form a 5- or 6-membered heterocyclyl ring, R 5a However, C 6 -C 20 Aryl and C 1 -C 20 selected from the group consisting of heteroaryl, In the formula, the asterisk * indicates the binding site of L, and R 1 ,R 2 ,R 3 , and R 4 One of them is bonded to L, L is -C(=O)-PEG-, -C(=O)-PEG-C(=O)N(R 6 )-(C 1 -C 12 alkyldiyl)-C(=O)-Gluc-, -C(=O)-PEG-O-, -C(=O)-PEG-O-C(=O)-, -C(=O)-PEG-C(=O)-, -C(=O)-PEG-C(=O)-PEP-, -C(=O)-PEG-N(R 6 )-, -C(=O)-PEG-N(R 6 )-C(=O)-, -C(=O)-PEG-N(R 6 )-PEG-C(=O)-PEP-, -C(=O)-PEG-N + (R 6 ) 2 -PEG-C(=O)-PEP-, -C(=O)-PEG-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-, -C(=O)-PEG-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)N(R 6 )C(=O)-(C 2 -C 5 monoheterocyclyldiyl)-, -C(=O)-PEG-SS-(C 1 -C 12 alkyldiyl)-OC(=O)-, -C(=O)-PEG-SS-(C 1 -C 12 alkyldiyl)-C(=O)-, -C(=O)-(C 1 -C 12 alkyldiyl)-C(=O)-PEP-, -C(=O)-(C 1 -C 12 alkyldiyl)-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-, -C(=O)-(C 1 -C 12 alkyldiyl)-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-N(R 5 )-C(=O), -C(=O)-(C 1 -C 12 alkyldiyl)-C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-N(R 6 )C(=O)-(C2 -C 5 monoheterocyclyldiyl)-, -Succinimidyl-(CH 2 ) m -C(=O)N(R 6 )-PEG-, -Succinimidyl-(CH 2 ) m -C(=O)N(R 6 )-PEG-C(=O)N(R 6 )-(C 1 -C 12 alkyldiyl)-C(=O)-Gluc-, -Succinimidyl-(CH 2 ) m -C(=O)N(R 6 )-PEG-O-, -Succinimidyl-(CH 2 ) m -C(=O)N(R 6 )-PEG-O-C(=O)-, -Succinimidyl-(CH 2 ) m -C(=O)N(R 6 )-PEG-C(=O)-, -Succinimidyl-(CH 2 ) m -C(=O)N(R 6 )-PEG-N(R 5 )-, -Succinimidyl-(CH 2 ) m -C(=O)N(R 6 )-PEG-N(R 5 )-C(=O)-, -Succinimidyl-(CH 2 ) m -C(=O)N(R 6 )-PEG-C(=O)-PEP-, -Succinimidyl-(CH 2 ) m -C(=O)N(R 6 )-PEG-SS-(C 1 -C 12 alkyldiyl)-OC(=O)-, -Succinimidyl-(CH 2 ) m -C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)-, -Succinimidyl-(CH 2 ) m -C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)N(R 6 )C(=O)-, and -Succinimidyl-(CH 2 ) m -C(=O)-PEP-N(R 6 )-(C 1 -C 12 alkyldiyl)N(R 6 )C(=O)-(C 2 -C 5 a linker selected from the group consisting of monoheterocyclyldiyl)-; R 6 but independently, H or C 1 -C 6 is alkyl, PEG has the formula:-(CH 2 CH 2 O) n -(CH 2 ) m -, m is an integer from 1 to 5, n is an integer from 2 to 50, Gluc has the formula: [ka] has PEP has the formula: [ka] , where AA is independently selected from natural or non-natural amino acid side chains, or one or more of AA and the adjacent nitrogen atom form a 5-membered proline amino acid. form, the wavy line indicates the bonding point, Cyc is optionally F, Cl, NO 2 , -OH, -OCH 3 , and C substituted with one or more groups selected from glucuronic acid having the structure: 6 -C 20 Aryldiyl and C 1 -C 20 selected from heteroaryldiyl, [ka] R 7 But -CH(R 8 )O-, -CH 2 -,-CH 2 N(R 8 )-, and -CH(R 8 )O-C(=O)-, where R 8 But, H, C 1 -C 6 Alkyl, C(=O)-C 1 -C 6 Alkyl, and -C(=O)N(R 9 ) 2 selected from, where R 9 But independently, H, C 1 -C 12 alkyl, and -(CH 2 CH 2 O) n -(CH 2 ) m -OH, where m is an integer from 1 to 5, n is an integer from 2 to 50, or two R 9 the groups together form a 5- or 6-membered heterocyclyl ring, y is an integer from 2 to 12, z is 0 or 1, Alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl are independently and optionally F, Cl, Br, I, -CN, -CH 3 ,-CH 2 CH 3 ,-CH=CH 2 , -C≡CH, -C≡CCH 3 ,-CH 2 CH 2 CH 3 , -CH(CH 3 ) 2 ,-CH 2 CH(CH 3 ) 2 ,-CH 2 OH, -CH 2 OCH 3 ,-CH 2 CH 2 OH, -C(CH 3 ) 2 OH, -CH(OH)CH(CH 3 ) 2 , -C(CH 3 ) 2 CH 2 OH, -CH 2 CH 2 S.O. 2 CH 3 ,-CH 2 OP(O)(OH) 2 ,-CH 2 F, -CHF 2 ,-CF 3 ,-CH 2 C.F. 3 ,-CH 2 CHF 2 , -CH(CH 3 )CN, -C(CH 3 ) 2 CN, -CH 2 CN, -CH 2 N.H. 2 ,-CH 2 NHSO 2 CH 3 ,-CH 2 NHCH 3 ,-CH 2 N(CH 3 ) 2 ,-CO 2 H,-COCH 3 ,-CO 2 CH 3 ,-CO 2 C(CH 3 ) 3 , -COCH(OH)CH 3 ,-CONH 2 ,-CONHCH 3 ,-CON(CH 3 ) 2 , -C(CH 3 ) 2 CONH 2 , -NH 2 , -NHCH 3 , -N(CH 3 ) 2 ,-NHCOCH 3 , -N(CH 3 )COCH 3 ,-NHS(O) 2 CH 3 , -N(CH 3 )C(CH 3 ) 2 CONH 2 , -N(CH 3 )CH 2 CH 2 S(O) 2 CH 3 , -NHC(=NH)H, -NHC(=NH)CH 3 , -NHC(=NH)NH 2 ,-NHC(=O)NH 2 ,-NO 2 , =O, -OH, -OCH 3 ,-OCH2 CH 3 ,-OCH 2 CH 2 OCH 3 ,-OCH 2 CH 2 OH, -OCH 2 CH 2 N(CH 3 ) 2 , -O(CH 2 CH 2 O) n -(CH 2 ) m C.O. 2 H, -O(CH 2 CH 2 O) n H, -OCH 2 F,-OCHF 2 ,-OCF 3 , -OP(O)(OH) 2 , -S(O) 2 N(CH 3 ) 2 ,-SCH 3 , -S(O) 2 CH 3 , and -S(O) 3 Said immunoconjugate, or a pharmaceutically acceptable salt thereof, is substituted with one or more groups independently selected from H.

[0106] Exemplary embodiments of the immunoconjugate of Formula I include that the antibody is selected from trastuzumab and pertuzumab, or biosimilars or biobetters thereof.

[0107] Exemplary embodiments of the immunoconjugate of Formula I include that Het is selected from the group consisting of pyridyldiyl, pyrimidyldiyl, pyrazolyldiyl, piperazinyldiyl, piperidinyldiyl, and pyrazinyldiyl.

[0108] An exemplary embodiment of the immunoconjugate of formula I is 1 is a combination and R 1 includes that is H.

[0109] An exemplary embodiment of the immunoconjugate of formula I is 2 is a combination and R 2 However, C 1 -C 8 Including being alkyl.

[0110] An exemplary embodiment of the immunoconjugate of formula I is 2 and X 3 are each a combination and R 2 and R 3 But independently, C 1 -C 8 Alkyl, -O-(C 1 -C 12 alkyl), -(C 1 -C 12 alkyldiyl)-OR 5 , -(C 1 -C 8 alkyldiyl)-N(R 5 )CO 2 R 5 , -(C 1 -C 12 alkyl)-OC(O)N(R 5 ) 2 , -O-(C 1 -C 12 alkyl)-N(R 5 )CO 2 R 5 , and -O-(C 1 -C 12 alkyl)-OC(O)N(R 5 ) 2 including being selected from.

[0111] An exemplary embodiment of the immunoconjugate of Formula I is R 2 However, C 1 -C 8 is alkyl and R 3 But -(C 1 -C 8 alkyldiyl)-N(R 5 )CO 2 R 5 including being.

[0112] An exemplary embodiment of the immunoconjugate of Formula I is R 2 But -CH 2 CH 2 CH 3 and R 3 But -CH 2 CH 2 CH 2 N.H.C.O. 2 (t-Bu), -OCH 2 CH 2 N.H.C.O. 2 (cyclobutyl), and -CH 2 CH 2 CH 2 N.H.C.O. 2 (cyclobutyl).

[0113] An exemplary embodiment of the immunoconjugate of Formula I is R 2 and R 3 , each independently, -CH 2 CH 2 CH 3 ,-OCH 2 CH 3 ,-OCH 2 C.F. 3 ,-CH 2 CH 2 C.F. 3 ,-OCH 2 CH 2 OH, and -CH 2 CH 2 CH 2 Including being selected from OH.

[0114] An exemplary embodiment of the immunoconjugate of Formula I is R 2 and R 3 , respectively -CH 2 CH 2 CH 3 including being.

[0115] An exemplary embodiment of the immunoconjugate of Formula I is R 2 But -CH 2 CH 2 CH 3 and R 3 But -OCH 2 CH 3 including being.

[0116] An exemplary embodiment of the immunoconjugate of formula I is 3 -R 3 but, [ka] including being selected from the group consisting of.

[0117] An exemplary embodiment of the immunoconjugate of formula I is 4 is a combination and R 4 includes that is H.

[0118] An exemplary embodiment of the immunoconjugate of Formula I is R 1 is connected to L.

[0119] An exemplary embodiment of the immunoconjugate of Formula I is R 2 or R 3 is connected to L.

[0120] An exemplary embodiment of the immunoconjugate of formula I is 3 -R 3 -L is [ka] where the wavy line indicates the point of attachment to N.

[0121] An exemplary embodiment of the immunoconjugate of Formula I is R 4 However, C 1 -C 12 Including being alkyl.

[0122] An exemplary embodiment of the immunoconjugate of Formula I is R 4 But -(C 1 -C 12 alkyldiyl)-N(R 5 )-*, where the asterisk * indicates the binding site of L.

[0123] Exemplary embodiments of the immunoconjugate of Formula I include that L is -C(=O)-PEG- or -C(=O)-PEG-C(=O)-.

[0124] Exemplary embodiments of the immunoconjugate of Formula I include that L is attached to cysteine ​​thiol of the antibody.

[0125] Exemplary embodiments of the immunoconjugate of Formula I include, for PEG, m is 1 or 2 and n is an integer from 2 to 10.

[0126] An exemplary embodiment of the immunoconjugate of Formula I includes that n is 10 for PEG.

[0127] An exemplary embodiment of the immunoconjugate of formula I is where L comprises PEP, PEP is a dipeptide, and the formula: [ka] including having.

[0128] An exemplary embodiment of the immunoconjugate of formula I is AA 1 and A.A.2 but independently, H, -CH 3 , -CH(CH 3 ) 2 ,-CH 2 (C 6 H 5 ), -CH 2 CH 2 CH 2 CH 2 N.H. 2 ,-CH 2 CH 2 CH 2 NHC(NH)NH 2 ,-CHCH(CH 3 )CH 3 ,-CH 2 S.O. 3 H, and -CH 2 CH 2 CH 2 NHC(O)NH 2 selected from or AA 1 and A.A. 2 to form a five-membered proline amino acid.

[0129] An exemplary embodiment of the immunoconjugate of formula I is AA 1 is -CH(CH 3 ) 2 and A.A. 2 But -CH 2 CH 2 CH 2 NHC(O)NH 2 including being.

[0130] An exemplary embodiment of the immunoconjugate of formula I is AA 1 and A.A. 2 but independently, GlcNAc aspartate, -CH 2 S.O. 3 H, and -CH 2 OPO 3 Including being selected from H.

[0131] Exemplary embodiments of the immunoconjugate of formula I are those in which PEP has the formula: [ka] , where AA 1 and A.A. 2 independently selected from the side chains of naturally occurring amino acids.

[0132] An exemplary embodiment of the immunoconjugate of formula I is where L comprises PEP, PEP is a tripeptide, and the formula: [ka] including having.

[0133] An exemplary embodiment of the immunoconjugate of formula I is where L comprises PEP, PEP is a tetrapeptide, and the formula: [ka] including having.

[0134] An exemplary embodiment of the immunoconjugate of Formula I is: A.A. 1 is selected from the group consisting of Abu, Ala, and Val; A.A. 2 is selected from the group consisting of Nle(O-Bzl), Oic, and Pro, A.A. 3 But Ala and Met(O) 2 selected from the group consisting of A.A. 4 But Oic, Arg(NO 2 ), Bpa, and Nle(O-Bzl).

[0135] Exemplary embodiments of the immunoconjugate of Formula I include L comprises PEP, and PEP is Ala-Pro-Val, Asn-Pro-Val, Ala-Ala-Val, Ala-Ala-Pro-Ala, Ala-Ala-Pro-Val, and Ala-Ala-Pro-Nva.

[0136] An exemplary embodiment of the immunoconjugate of Formula I is where L comprises PEP, and PEP has the structure: [ka] including being selected from.

[0137] Exemplary embodiments of the immunoconjugate of Formula I have the structure: [ka] In the formula, the wavy line is R 5 including indicating a bond to.

[0138] Formula Ia: [ka] An exemplary embodiment of an immunoconjugate of formula I having:

[0139] An exemplary embodiment of the immunoconjugate of formula Ia is 4 is a combination and R 4 includes that is H.

[0140] Exemplary embodiments of the immunoconjugate of Formula Ia include that Het is selected from the group consisting of pyridyldiyl, pyrimidyldiyl, pyrazolyldiyl, piperazinyldiyl, piperidinyldiyl, and pyrazinyldiyl.

[0141] An exemplary embodiment of the immunoconjugate of formula Ia is 2 and X 3 are each a bond and R 2 and R 3 But independently, C 1 -C 8 Alkyl, -O-(C 1 -C 12 alkyl), -(C1-C 12 alkyldiyl)-OR 5 , -(C 1 -C 8 alkyldiyl)-N(R 5 )CO 2 R 5 , and -O-(C 1 -C 12 alkyl)-N(R 5 )CO 2 R 5 including being selected from.

[0142] Formula Ib~Ii: [ka] [ka] An exemplary embodiment of an immunoconjugate of Formula Ia selected from Ii.

[0143] An exemplary embodiment of the immunoconjugate of formula Ia is 2 and X 3 are each a bond and R 2 and R 3 But independently, C 1 -C 8 Alkyl, -O-(C 1 -C 12 alkyl), -(C1-C 12 alkyldiyl)-OR 5 , -(C 1 -C 8 alkyldiyl)-N(R 5 )CO 2 R 5 , and -O-(C 1 -C 12 alkyl)-N(R 5 )CO 2 R 5 including being selected from.

[0144] An exemplary embodiment of the immunoconjugate of formula Ia is 2 and X 3 are each a combination and R 2 However, C 1 -C 8 is alkyl and R 3 But -O-(C 1 -C 12 alkyl) and -O-(C 1 -C 12 alkyl)-N(R 5 )CO 2 R 5 including being selected from.

[0145] The invention includes all reasonable combinations and permutations of features of the Formula I embodiments.

[0146] In certain embodiments, immunoconjugate compounds of the invention include those that have immunostimulatory activity. The antibody-drug conjugate of the invention selectively delivers an effective dose of 8-Het-2-aminobenzazepine drug to tumor tissue, thereby providing a higher selectivity (ie, lower effective doses) can be achieved while increasing the therapeutic index ("therapeutic window").

[0147] Drug loading is expressed as p, which is the number of Hx moieties per antibody in the formula I immunoconjugate. Drug (Hx) loading can range from 1 to about 8 drug moieties (D) per antibody. The immunoconjugate of Formula I comprises a mixture or collection of antibodies conjugated with a range of from 1 to about 8 drug moieties. In some embodiments, the number of drug moieties that can be conjugated to an antibody is limited by the number of reactive or available amino acid side chain residues, such as lysine and cysteine. In some embodiments, free cysteine ​​residues are introduced into antibody amino acid sequences by the methods described herein. In such embodiments, p may be 1, 2, 3, 4, 5, 6, 7, or 8, or ranges thereof, such as 1-8 or 2-5. In any such embodiment, p and n are equal (ie, p=n=1, 2, 3, 4, 5, 6, 7, or 8, or some range therebetween). Exemplary immunoconjugates of Formula I include, but are not limited to, antibodies with 1, 2, 3, or 4 engineered cysteine ​​amino acids (Lyon, R. et al. (2012) Methods in Enzym.502:123-138). In some embodiments, one or more free cysteine ​​residues are already present in the antibody to form an intrachain disulfide bond without the use of manipulation, in which case the existing free cysteine ​​residues are used to form an intrachain disulfide bond. , the antibody may be conjugated to a drug. In some embodiments, the antibody is exposed to reducing conditions prior to conjugation of the antibody to generate one or more free cysteine ​​residues.

[0148] In some immunoconjugates, p may be limited by the number of binding sites on the antibody. For example, if the linkage is a cysteine ​​thiol, as in certain exemplary embodiments described herein, the antibody may have only one or a limited number of cysteine ​​thiol groups, or The drug may have only one or a limited number of sufficiently reactive thiol groups to which it may be attached. In other embodiments, one or more lysine amino groups in the antibody may be available and reactive for conjugation with an Hx-linker compound of Formula II. In certain embodiments, higher drug loading, eg, p>5, can cause aggregation, insolubility, toxicity, or loss of cell permeability of certain antibody-drug conjugates. In certain embodiments, the average drug loading of the immunoconjugate ranges from 1 to about 8, about 2 to about 6, or about 3 to about 5. In certain embodiments, antibodies are subjected to denaturing conditions to reveal reactive nucleophilic groups such as lysine or cysteine.

[0149] The loading of the immune complex (drug / antibody ratio) can be controlled in different ways, e.g. by (i) limiting the molar excess of Hx-linker intermediate compound over the antibody and (ii) limiting the conjugation reaction time or temperature. and (iii) partial or limited reductive denaturation conditions for optimized antibody reactivity.

[0150] It is to be understood that when two or more nucleophilic groups of an antibody react with a drug, the resulting product is a mixture of immunoconjugated compounds with a distribution of one or more drug moieties attached to the antibody. . The average number of drugs per antibody can be calculated from the mixture by a dual ELISA antibody assay that is specific for the antibody and specific for the drug. Individual immune complex molecules can be identified in mixtures by mass spectrometry and separated by HPLC, e.g. hydrophobic interaction chromatography (e.g. McDonagh et al. (2006) Prot. Engr. Design & Selection19(7) ):299-307, Hamblett et al. (2004) Clin. Cancer Res.10:7063-7070, Hamblett, K.J., et al. “Effect of drug loading on the pharmacology, pharmacokinetics, and toxicity of an anti-CD30 antibody -drug conjugate,"Abstract No.624,American Association for Cancer Research,2004 Annual Meeting,March27-31,2004,Proceedings of the AACR,Volume 45,March 2004,Alley, S.C.,et al."Controlling the location of drug attachment in antibody-drug conjugates,” Abstract No. 627, American Association for Cancer Research, 2004 Annual Meeting, March 27-31, 2004, Proceedings of the AACR, Volume 45, March 2004). In certain embodiments, homogeneous immune complexes with a single loading value can be isolated from a conjugation mixture by electrophoresis or chromatography.

[0151] Exemplary embodiments of the immunoconjugates of formula I are selected from Table 3 immunoconjugates. In vitro evaluation of immune complex activity was performed according to the method of Example 203. [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4]

[0152] Composition of immune complexes The invention provides compositions, e.g., pharmaceutical or pharmacologically acceptable carriers, comprising a plurality of immunoconjugates described herein, and optionally a carrier thereof, e.g. provides a composition or formulation that is acceptable for The immune complexes may be the same or different in composition, i.e., the composition may include immune complexes with the same number of adjuvants linked to the same position on the antibody construct, and / or different adjuvants on the antibody construct. Immune complexes with the same number of Hx adjuvants linked to positions, different numbers of adjuvants linked to the same positions on the antibody construct, or different numbers of adjuvants linked to different positions on the antibody construct. can include immune complexes with.

[0153] In an exemplary embodiment, the composition comprising an immunoconjugate compound comprises a mixture of immunoconjugate compounds, and the average drug (Hx) load per antibody in the mixture of immunoconjugate compounds is from about 2 to about 5 It is.

[0154] The immunoconjugate compositions of the invention can have an average adjuvant to antibody construct ratio (DAR) of about 0.4 to about 10. Those skilled in the art will recognize that the number of 8-Het-2-aminobenzazepine adjuvants conjugated to an antibody construct can vary from immunoconjugate to immunoconjugate in a composition comprising multiple immunoconjugates of the invention. Therefore, the adjuvant-to-antibody construct (eg, antibody) ratio can be measured as an average, which can be referred to as the drug-to-antibody ratio (DAR). The adjuvant-to-antibody construct (eg, antibody) ratio can be assessed by any suitable means, many of which are known in the art.

[0155] The average number of adjuvant sites per antibody (DAR) in the preparation of immune complexes from conjugation reactions can be characterized by conventional means such as mass spectrometry, ELISA assays, and HPLC. Quantitative distribution of immune complexes in the composition in terms of p can also be determined. In some cases, the separation, purification, and characterization of homogeneous immune complexes for which p is a particular value from other drug-loaded immune complexes is accomplished by means such as reversed-phase HPLC or electrophoresis. can be achieved.

[0156] In some embodiments, the composition further comprises one or more pharmaceutically or pharmacologically acceptable excipients. For example, the immunoconjugates of the invention can be formulated for parenteral administration, such as IV administration or administration into a body cavity or lumen of an organ. Alternatively, the immune complex can be injected intratumorally. Injectable compositions generally include a solution of the immune complex dissolved in a pharmaceutically acceptable carrier. Among the acceptable vehicles and solvents that may be employed are water and isotonic solutions of one or more salts such as sodium chloride, eg, Ringer's solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid can similarly be used in the preparation of injectables. These compositions are desirably sterile and generally free of undesirable materials. These compositions can be sterilized by conventional, well-known sterilization techniques. The compositions may contain pharmaceutical additives as necessary to approximate physiological conditions, such as pH adjusting agents and buffers, toxicity modifiers, such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, etc. It may contain acceptable auxiliary substances.

[0157] The composition can contain any suitable concentration of immunoconjugate. The concentration of immunoconjugate in the composition can vary widely and is selected based primarily on fluid volume, viscosity, body weight, etc., depending on the particular mode of administration chosen and the needs of the patient. In certain embodiments, the concentration of the immunoconjugate in the injectable solution formulation ranges from about 0.1% (w / w) to about 10% (w / w).

[0158] How to treat cancer with immune complexes The invention provides methods for treating cancer. The method comprises administering a therapeutically effective amount of an immunoconjugate described herein (e.g., as a composition described herein) to a subject in need thereof, e.g., having cancer, for treatment of cancer. including administering to a subject in need. The method includes administering a therapeutically effective amount of an immunoconjugate (IC) selected from Table 3a and Table 3b.

[0159] It is contemplated that the immunoconjugates of the invention can be used, for example, to treat a variety of hyperproliferative diseases or disorders characterized by overexpression of tumor antigens. Exemplary hyperproliferative disorders include solid tumors, benign or malignant, and hematological disorders such as leukemias and lymphoid malignancies.

[0160] In another aspect, an immunoconjugate for use as a pharmaceutical is provided. In certain embodiments, the invention provides an immunoconjugate for use in a method of treating an individual comprising administering to the individual an effective amount of the immune conjugate. In one such embodiment, the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent, eg, as described herein.

[0161] In a further aspect, the invention provides the use of an immunoconjugate in the manufacture or preparation of a medicament. In one embodiment, the pharmaceutical agent is for the treatment of cancer, and the method includes administering to an individual with cancer an effective amount of the pharmaceutical agent. In one such embodiment, the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent, eg, as described herein.

[0162] Carcinoma is a malignant tumor derived from epithelial tissue. Epithelial cells line the external surfaces of the body, line the internal cavities, and form the lining of glandular tissue. Examples of carcinoma include adenocarcinoma (cancer that develops in glandular (secretory) cells such as breast cancer, pancreatic cancer, lung cancer, prostate cancer, stomach cancer, gastroesophageal cancer, and colon cancer), adrenocortical cancer, hepatocellular carcinoma, Renal cell carcinoma, ovarian cancer, carcinoma in situ, ductal carcinoma, breast cancer, basal cell carcinoma, squamous cell carcinoma, transitional cell carcinoma, colon cancer, nasopharyngeal carcinoma, polycystic renal cell carcinoma, oat cell carcinoma, large cell lung cancer , small cell lung cancer, non-small cell lung cancer, etc., but are not limited to these. Carcinomas may be found in the prostate, pancreas, colon, brain (usually as secondary metastases), lungs, breasts, and skin. In some embodiments, the method for treating non-small cell lung cancer contains an antibody construct capable of binding to HER2 (e.g., trastuzumab, pertuzumab, biosimilars thereof, or biobetters thereof) including administering an immune complex.

[0163] Soft tissue tumors are a highly diverse group of rare tumors derived from connective tissue. Examples of soft tissue tumors include alveolar soft tissue sarcoma, angiomatoid fibrous histiocytoma, chondromuscular oxidative fibroma, skeletal chondrosarcoma, extraskeletal myxoid chondrosarcoma, clear cell sarcoma, and desmoplastic small round cell Tumor, dermatofibrosarcoma protuberance, endometrial stromal tumor, Ewing sarcoma, fibromatosis (desmoid), infantile fibrosarcoma, gastrointestinal stromal tumor, giant cell tumor of bone, tenosynovial giant cell tumor, inflammatory muscle fiber Blastic tumor, uterine leiomyoma, leiomyosarcoma, lipoblastoma, typical lipoma, spindle cell or pleomorphic lipoma, atypical lipoma, chondrolipoma, well-differentiated liposarcoma, myxoid / round cell liposarcoma, pleomorphic liposarcoma, myxoid malignant fibrous histiocytoma, high-grade malignant fibrous histiocytoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, mesothelioma, neuroblastoma, Osteochondroma, osteosarcoma, primitive neuroectodermal tumor, alveolar rhabdomyosarcoma, embryonic rhabdomyosarcoma, benign or malignant schwannoma, synovial sarcoma, Evan's tumor, nodular fasciitis, desmoid type Fibromatosis, solitary fibrous tumor, dermatofibrosarcoma protuberans (DFSP), angiosarcoma, epithelioid hemangioendothelioma, tenosynovial giant cell tumor (TGCT), pigmented villonodular synovitis (PVNS), fibrosis Sexual dysplasia, myxofibrosarcoma, fibrosarcoma, synovial sarcoma, malignant peripheral nerve sheath tumors, neurofibromas, pleomorphic adenomas of soft tissues, as well as fibroblasts, myofibroblasts, histiocytes, vascular cells / endothelium cells, and neoplasms derived from nerve sheath cells.

[0164] Sarcomas are a rare type of cancer that arises in cells of mesenchymal origin, such as bones or soft tissues of the body, including cartilage, fat, muscle, blood vessels, fibrous tissue, or other connective or supporting tissues. Different types of sarcoma are based on where the cancer forms. For example, osteosarcoma forms in bone, liposarcoma in fat, and rhabdomyosarcoma in muscle. Examples of sarcomas include Askin's tumor, botryoides sarcoma, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant Schwannoma, osteosarcoma, and soft tissue sarcomas (e.g., alveolar soft tissue sarcoma, angiosarcoma, cystic sarcoma, fjord fibrous sarcoma) DFSP, desmoid tumor, deplastic small round cell tumor, epithelial sarcoma, extraosseous chondrosarcoma, extraosseous osteosarcoma, fibrous sarcoma, peptic gastrointestinal stromal tumor (GIST), vascular cancer, angiosarcoma ( (more commonly referred to as "angiosarcoma"), Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, synovial sarcoma, and undifferentiated tumor. plastic sarcoma), but are not limited to these.

[0165] A metamorphoma can contain several different types of tissue, including, for example, hair, muscle, and bone (e.g., any and / or all of the three germ layers: endoderm, mesoderm, and ectoderm). is a type of germ cell tumor that can include tissue derived from. Deformities occur most frequently on the ovaries in women, testicles in men, and the coccyx in children.

[0166] Melanoma is a form of cancer that develops in melanocytes (cells that make the pigment melanin). Melanoma can develop in moles (cutaneous melanoma) but can also develop in other pigmented tissues such as the eyes or intestines.

[0167] Merkel cell carcinoma is a rare type of skin cancer that usually appears as flesh-colored or bluish-red nodules on the face, head, or neck. Merkel cell carcinoma is also called neuroendocrine carcinoma of the skin. In some embodiments, the method for treating Merkel cell carcinoma comprises an immunotherapy comprising an antibody construct capable of binding to HER2 (e.g., trastuzumab, pertuzumab, biosimilars thereof, or biobetters thereof). and administering the complex. In some embodiments, the Merkel cell carcinoma has metastasized at the time the administration occurs.

[0168] Leukemia is a cancer that begins in blood-forming tissues such as the bone marrow, where large numbers of abnormal blood cells are produced and enter the bloodstream. For example, leukemia can originate from bone marrow-derived cells that normally mature in the bloodstream. Leukemias are named by the rate of onset and progression of the disease (eg, acute versus chronic) and the type of white blood cells affected (eg, bone marrow versus lymph). Myeloid leukemia is also called myeloid or myeloblastic leukemia. Lymphocytic leukemia is also called lymphoblastic or lymphocytic leukemia. Lymphocytic leukemia cells can collect in lymph nodes, which can become swollen. Examples of leukemias include, but are not limited to, acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), and chronic lymphocytic leukemia (CLL).

[0169] Lymphoma is a cancer that affects cells of the immune system. For example, lymphomas can be derived from bone marrow-derived cells that normally mature in the lymphatic system. There are two basic categories of lymphoma. One category of lymphoma is Hodgkin lymphoma (HL), which is characterized by the presence of a type of cell called Reed-Sternberg cells. Currently, six types of HL are recognized. Examples of Hodgkin's lymphoma include nodular sclerosis classic Hodgkin's lymphoma (CHL), mixed cellular CHL, lymphopenic CHL, lymphocyte-rich CHL, and nodular lymphocyte-predominant HL.

[0170] Another category of lymphoma is non-Hodgkin's lymphoma (NHL), which includes a large and diverse group of cancers of immune system cells. Non-Hodgkin's lymphoma is further divided into cancers with an indolent (slow-growing) course and cancers with an rapid (rapid-growing) course. Currently, 61 types of NHL are recognized. Examples of non-Hodgkin's lymphoma include AIDS-related lymphoma, anaplastic large cell lymphoma, angioimmunoblastic lymphoma, blastic NK cell lymphoma, Burkitt's lymphoma, Burkitt-like lymphoma (small non-cleaved cell lymphoma), chronic Lymphocytic leukemia / small lymphocytic lymphoma, cutaneous T-cell lymphoma, diffuse large B-cell lymphoma, enteropathic T-cell lymphoma, follicular lymphoma, hepatosplenic gamma delta T-cell lymphoma, T-cell leukemia, lymphoma Blastic lymphoma, mantle cell lymphoma, marginal zone lymphoma, nasal T-cell lymphoma, childhood lymphoma, peripheral T-cell lymphoma, primary central nervous system lymphoma, transformed lymphoma, therapy-related T-cell lymphoma, and Waldenstrees These include, but are not limited to, mummacroglobulinemia.

[0171] Brain tumors include any cancer of brain tissue. Examples of brain tumors include gliomas (e.g., glioblastomas, astrocytomas, oligodendrogliomas, ependymomas, etc.), meningiomas, pituitary adenomas, and vestibular schwannoma, primitive neuroectodermal tumors ( including, but not limited to, medulloblastoma).

[0172] The immunoconjugates of the invention can be used in therapy either alone or in combination with other agents. For example, the immunoconjugate may be administered with at least one additional therapeutic agent, such as a chemotherapeutic agent. Such combination therapy includes combined administration (where two or more therapeutic agents are included in the same or separate formulations), and separate administration, where administration of the immunoconjugate is administered in combination with the additional therapeutic agent and / or It may occur before, simultaneously with, and / or after administration of the adjuvant. Immunoconjugates can also be used in combination with radiation therapy.

[0173] The immunoconjugates of the invention (and any additional therapeutic agents) may be administered by any suitable means, including parenterally, intrapulmonally, and intranasally, as well as intralesional administration, if desired for topical treatment. obtain. Parenteral injections include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Dosing can be carried out by any suitable route, eg, by injection, such as intravenous or subcutaneous injection, depending in part on whether administration is brief or chronic. A variety of dosing schedules are contemplated herein, including, but not limited to, single or multiple administrations over various time points, bolus administration, and pulse infusion.

[0174] The immunoconjugate can be administered to a subject in need thereof in any therapeutically effective amount using any suitable dosing regimen, such as those utilized for labetuzumab, its biosimilars, and its biobetters. Ru. For example, the method can include administering the immune complex to provide a dose of about 100 ng / kg to about 50 mg / kg to the subject. Immunoconjugate doses can range from about 5 mg / kg to about 50 mg / kg, about 10 μg / kg to about 5 mg / kg, or about 100 μg / kg to about 1 mg / kg. The immunoconjugate dose can be about 100, 200, 300, 400, or 500 μg / kg. The immunoconjugate dose can be about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg. Immunoconjugate doses can also fall outside these ranges depending on the particular conjugate and the type and severity of cancer being treated. The frequency of administration can range from a single dose to multiple doses per week, or more frequently. In some embodiments, the immunoconjugate is administered from about 1 time per month to about 5 times per week. In some embodiments, the immunoconjugate is administered once a week.

[0175] In another aspect, the invention provides a method for preventing cancer. The method includes administering to a subject a therapeutically effective amount of an immunoconjugate (eg, as a composition as described above). In certain embodiments, the subject is susceptible to the particular cancer being prevented. For example, the method can include administering the immune complex to provide a dose of about 100 ng / kg to about 50 mg / kg to the subject. Immunoconjugate doses can range from about 5 mg / kg to about 50 mg / kg, about 10 μg / kg to about 5 mg / kg, or about 100 μg / kg to about 1 mg / kg. The immunoconjugate dose can be about 100, 200, 300, 400, or 500 μg / kg. The immunoconjugate dose can be about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg. Immunoconjugate doses can also fall outside these ranges depending on the particular conjugate and the type and severity of cancer being treated. The frequency of administration can range from a single dose to multiple doses per week, or more frequently. In some embodiments, the immunoconjugate is administered from about 1 time per month to about 5 times per week. In some embodiments, the immunoconjugate is administered once a week.

[0176] Some embodiments of the invention provide methods for treating cancer as described above, where the cancer is breast cancer. Breast cancer can arise from different areas of the breast, and a number of different types of breast cancer have been characterized. For example, the immunoconjugates of the invention may be used for ductal carcinoma in situ, invasive ductal carcinoma (e.g., tubular, medullary, mucinous, papillary, or cribriform carcinoma), lobular carcinoma in situ, invasive It can be used to treat other forms of breast cancer, such as lobular carcinoma, inflammatory breast cancer, and triple-negative (tests negative for estrogen receptors, progesterone receptors, and excess HER2 protein) breast cancer. In some embodiments, methods for treating breast cancer include antibody constructs that can bind to HER2, or tumors that overexpress HER2 (e.g., trastuzumab, pertuzumab, biosimilars, or biobetters thereof). comprising administering an immunoconjugate containing.

[0177] In some embodiments, the cancer is susceptible to pro-inflammatory responses induced by TLR7 and / or TLR8.

Example

[0178] Example L-5 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-[2-Amino-4- [ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrazol-1-yl]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2 Synthesis of ,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-5 [ka] tert-Butyl3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-(4,4,5,5-tetramethyl- Preparation of 1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanate, HxBzL-5a PPh 3 (1.35 g, 5.15 mmol, 1 eq.) and DEAD (0.89 g, 5.15 mmol, 0.94 mL, 1 eq.) were added at 0 °C, stirred at 25 °C for 0.5 h, then tert-butyl 3-[2- [2-[2-[2-[2-[2-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid Salt (3.02g, 5.15mmol, 1eq) was added and then stirred at 25°C for 16 hours. The reaction mixture was diluted with 20 mL of water and extracted with EtOAc (50 mL*3). Wash the mixed organic layer with brine (20mL*3) and add Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate=50 / 1 to ethyl acetate:MeOH=10:1) to obtain HxBzL-5a (3.5 g, 4.59 mmol, yield 89.04%) as a yellow oil.

[0179] tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-[2-amino-4-[ethoxy(propyl)] Preparation of HxBzL-5c HxBzL-2a (625 mg, 819 umol, 2.5 eq.), 2-amino-8-bromo-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide, HxBzL-2b (120 mg, 328 umol, 1 eq.), Na in water (0.3mL) 2 C.O. 3 (69.5 mg, 655 umol, 2 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride, Pd(dppf)Cl in DMF (3 mL). 2 (23.9 mg, 32.8 umol, 0.1 eq.) was degassed and then N 2 The mixture was heated to 120°C for 5 hours. The mixture was filtered, concentrated under reduced pressure, and the residue was subjected to preparative HPLC (TFA conditions, column: Phenomenex luna C18 250*50mm*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 35% ~65%, 10 min) to give HxBzL-5c (300 mg, 290 umol, 88.4% yield, TFA) as a yellow solid.

[0180] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-[2-amino-4-[ethoxy(propyl)carbamoyl]- Preparation of 3H-1-benzazepin-8-yl]pyrazol-1-yl]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-5d To a solution of HxBzL-5c (300 mg, 325 umol, 1 eq.) in water (3 mL) and MeCN (0.5 mL) was added HCl (12M, 407 uL, 15 eq.) and then stirred at 80.degree. C. for 0.5 h. The mixture was concentrated under reduced pressure to give HxBzL-5d (200 mg, 222 umol, 68.1% yield, HCl) as a colorless oil.

[0181] Preparation of HxBzL-5 HxBzL-5d (80.0 mg, 88.7 umol, 1 eq., HCl) and sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (119 mg, 443 umol) in DCM (1 mL) and DMA (1 mL). , 5 eq.) was added 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, EDCI (84.9 mg, 443 umol, 5 eq.) and then stirred at 25° C. for 0.5 h. The mixture was filtered, concentrated under reduced pressure, and the residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 25 ~ 50%, 8 min) to give HxBzL-5 (30 mg, 24.8 umol, 28.01% yield, TFA) as a yellow oil. 1 H NMR (400MHz, MeOD) δ 8.20 (s, 1H), 7.93 (s, 1H), 7.65-7.61 (m, 1H), 7.59 (s, 1H), 7.55-7.52 (m, 1H), 7.40 (s , 1H), 4.36 (t, J = 4.8 Hz, 2H), 3.96 (q, J = 7.2 Hz, 2H), 3.89-3.82 (m, 4H), 3.74 (t, J = 7.2 Hz, 2H), 3.63 -3.52 (m, 36H), 3.42 (s, 2H), 2.95 (t, J = 5.6 Hz, 2H), 1.76 (sxt, J = 7.2 Hz, 2H), 1.20 (t, J = 7.2 Hz, 3H) , 0.99 (t, J = 7.6 Hz, 3H).LC / MS[M+H]1094.4 (calculated value), LC / MS[M+H]1094.3 (observed value).

[0182] Example L-7 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[4-[5-[2-amino -4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]piperazin-1-yl]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-7 [ka] 2-Amino-N-ethoxy-N-propyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3H-1-benzazepine-4-carboxamide, HxBz -Preparation of 4b 2-Amino-8-bromo-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide, HxBz-4a (0.5 g, 1.37 mmol, 1 eq.) in dioxane (10 mL), 4,4, 5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (520 mg, 2.05 mmol, 1.5 eq.) , Pd(dppf)Cl 2 (99.9 mg, 137 umol, 0.1 eq), KOAc (335 mg, 3.41 mmol, 2.5 eq) in N 2 The mixture was stirred at 100° C. for 1 hour. The crude HxBz-4b was used in the next step without purification and was obtained (564 mg, 1.36 mmol, 99.96% yield) as a black liquid.

[0183] tert-Butyl 4-[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin--8-yl]pyrimidin-2-yl]piperazine-1-carboxylate, HxBz-4 preparation of HxBz-4b (0.45 g, 1.09 mmol, 1 eq.) in dioxane (4 mL) and water (0.5 mL), Pd(dppf)Cl 2 (39.8mg, 54.4umol, 0.05eq), K 2 C.O. 3 (376 mg, 2.72 mmol, 2.5 eq.), tert-butyl 4-(5-bromopyrimidin-2-yl)piperazine-1-carboxylate (374 mg, 1.09 mmol, 1 eq.) in N 2 The mixture was stirred at 100° C. for 1 hour. The mixture was concentrated to remove dioxane and the residue was diluted with EtOAc (10 mL) and water (5 mL). organic layer with Na 2 S.O. 4 Dry on top and concentrate to give a residue. The residue was subjected to column chromatography (SiO 2 , petroleum ether / ethyl acetate=1 / 0~0 / 1, then EA:MeOH=1.5:1), then preparative HPLC, column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1% TFA)-ACN], B%: 30% to 55%, 8 min) to obtain HxBz-4 (0.35 g, 637 umol, yield 58.5%) as a brown oil. 1 H NMR (400MHz, MeOD) δ8.74 (s, 2H), 7.72-7.63 (m, 2H), 7.60 (d, J = 1.6 Hz, 1H), 7.45 (s, 1H), 3.99 (q, J = 7.2 Hz, 2H), 3.93-3.88 (m, 4H), 3.77 (t, J = 7.2 Hz, 2H), 3.60-3.51 (m, 4H), 3.43 (s, 2H), 1.80-1.75 (m, 2H) ), 1.51 (s, 9H), 1.22 (t, J = 7.2 Hz, 3H), 1.02 (t, J = 7.2 Hz, 3H).LC / MS[M+H]550.3 (calculated value), LC / MS [M+H]550.2 (observed value).

[0184] Preparation of 2-amino-N-ethoxy-8-(2-piperazin-1-ylpyrimidin-5-yl)-N-propyl-3H-1-benzazepine-4-carboxamide, HxBz-3 To a mixture of HxBz-4 (20 mg, 36.4 umol, 1 eq) in DCM (5 mL) was added HCl / EtOAc (4M, 5 mL, 550 eq) and stirred at 25 °C for 0.5 h. The mixture was concentrated to give HxBz-3 (10.5 mg, 21.4 umol, 58.9% yield, 99.233% purity, HCl) as a white solid. 1 H NMR (400MHz, MeOD) δ8.70 (s, 2H), 7.65-7.47 (m, 3H), 7.32 (s, 1H), 4.14-3.96 (m, 4H), 3.86 (q, J = 7.2 Hz, 2H), 3.64 (t, J = 7.2 Hz, 2H), 3.31 (s, 2H), 3.25-3.21 (m, 4H), 1.71-1.62 (m, 2H), 1.08(t, J = 7.2 Hz, 3H ), 0.89 (t, J = 7.2 Hz, 3H).LC / MS[M+H]450.3 (calculated value), LC / MS[M+H]450.1 (observed value).

[0185] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[1[3-[2-amino-4-[3-(tert -butoxycarbonylamino)propyl-ethoxy-carbamoyl]-3H-1-benzazepin-8-yl]phenyl]sulfonylazetidin-3-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy] Preparation of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-7a A mixture of HxBz-3 (110 mg, 176 umol, 1 eq.) in DMF (3 mL) was added with DIEA (63.5 mg, 491 umol, 2.8 eq.) and 3-[2-[2-[2-[2-[2- [2-[2-[2-[2-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Ethoxy]propanoic acid (99.2 mg, 140 umol, 0.8 eq.) was added and then stirred at 25° C. for 0.5 h. The mixture was purified by preparative HPLC (column: Waters Xbridge Prep OBD C18 150*40mm*10um, mobile phase: [water (10mMNH 4 HCO 3 )-ACN], B%: 20%-55%, 8 min) to give HxBzL-7a (28 mg, 24 umol, 13.7% yield) as a yellow oil.

[0186] Preparation of HxBzL-7 HxBzL-7a (78 mg, 78.8 umol, 1 eq) and sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (106 mg, 394 umol, 5 EDCI (75.5 mg, 394 umol, 5 eq.) was added to the mixture of 75.5 mg, 394 umol, 5 eq.) and then stirred at 20° C. for 0.5 h. The mixture was concentrated to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 20%~40%, 10 minutes) to obtain HxBzL. -7 (39.8 mg, 26.4 umol, yield 33.5%, purity 95.944%, 2TFA) was obtained as a colorless oil. 1H NMR (400MHz, MeOD) δ8.75 (s, 2H), 7.76-7.55 (m, 3H), 7.45 (s, 1H), 4.02-3.73 (m, 16H), 3.68-3.58 (m, 36H), 3.37 (s, 2H), 2.99 (t, J = 6.0 Hz, 2H), 2.76 (t, J = 6.0 Hz, 2H), 1.85-1.74 (m, 2H), 1.25-1.20 (m, 3H), 1.02 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1218.5 (calculated value), LC / MS[M+H]1218.3 (observed value).

[0187] Example L-8 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy Synthesis of ]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-8 [ka] Preparation of 5-bromo-2-(bromomethyl)pyrimidine, HxBz-5b PPh 3 (499mg, 1.90mmol, 1.2eq) and CBr 4 (631mg, 1.90mmol, 1.2eq), N 2 Added all at once at 0°C. The mixture was stirred at 20°C for 10 hours. Water (10 mL) was added, the aqueous phase was extracted with ethyl acetate (10 mL*3), the combined organic phase was washed with brine (10 mL), anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 8 / 1) to obtain HxBz-5b (290 mg, 1.15 mmol, yield). 72.4%) was obtained as a white solid. 1 H NMR (400 MHz, CDCl 3 ) δ8.81 (s, 2H), 4.59 (s, 2H).

[0188] Preparation of tert-butyl N-[(5-bromopyrimidin-2-yl)methyl]-N-tert-butoxycarbonyl-carbamate, HxBz-5c Cs 2 C.O. 3 (562mg, 1.73mmol, 1.5eq), N 2 was added portionwise at 20° C. and the mixture was stirred at 20° C. for 2.5 hours. Water (5 mL) was added, the aqueous phase was extracted with ethyl acetate (5 mL*3), the combined organic phase was washed with brine (5 mL), anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 5 / 1) to obtain HxBz-5c (350 mg, 901 umol, yield 78.3%) was obtained as a white solid. 1 H NMR (400 MHz, CDCl 3 ) δ8.74 (s, 2H), 5.01 (s, 2H), 1.48 (s, 18H).

[0189] Tert-Butyl N-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzaze-pin-8-yl]pyrimidin-2-yl]methyl]-N-tert-butoxy Preparation of carbonyl-carbamate, HxBz-5d Dioxane (10mL) and H 2 HxBz-5c (184 mg, 473 umol, 1.0 equiv.) and 2-amino-N-ethoxy-N-propyl-8-(4,4,5,5-tetramethyl-1,3,2- Pd(dppf)Cl was added to a mixture of dioxaborolan-2-yl)-3H-1-benzazepine-4-carboxamide (195 mg, 474 umol, 1.0 eq.). 2 ·CH 2 Cl 2 (19.3 mg, 23.7 umol, 0.05 equivalent) and K 2 C.O. 3 (163mg, 1.18mmol, 2.5eq), N 2 Degas the mixture by adding in one portion under N 2 The mixture was heated to 90°C for 2 hours. Dioxane (10 mL) was removed in vacuo, water (20 mL) was added, the aqueous phase was extracted with ethyl acetate (10 mL*3), the combined organic phases were washed with brine (10 mL) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100~200 mesh silica gel, petroleum ether / ethyl acetate = 10 / 1, 0 / 1 ~ ethyl acetate / methanol = 10 / 1) to obtain HxBz. -5d (280 mg, 470.83 umol, yield 99.35%) was obtained as a gray solid. 1 H NMR (400 MHz, MeOD) δ9.08 (s, 2H), 7.61 (s, 1H), 7.59 (d, J = 2.8 Hz, 2H), 7.38 (s, 1H), 5.08 (s, 2H), 3.98 (q, J = 7.2 Hz, 2H), 3.76 (t, J = 7.2 Hz, 2H), 1.83-1.75 (m, 2H), 1.47 (s, 18H), 1.20 (t, J = 7.2 Hz, 3H ), 1.02 (t, J = 7.2 Hz, 3H).

[0190] Preparation of 2-amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide, HxBz-5 To a solution of HxBz-5d (20.0 mg, 33.6 umol, 1.0 eq.) in EtOAc (5 mL) was added HCl / EtOAc (4M, 8.41 uL, 1.0 eq.) with N 2 was added in one portion at 20° C. and the mixture was stirred at 20° C. for 1 hour. The reaction mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1% TFA)-ACN], B%: 1%~30%, 8 min) to obtain HxBz -5 (6.2 mg, 9.84 umol, yield 29.2%, purity 98.8%, 2TFA) was obtained as a white solid. 1 H NMR (400 MHz, MeOD) δ9.22 (s, 2H), 7.82 (d, J = 2.0 Hz, 1H), 7.79-7.75 (m, 2H), 7.47 (s, 1H), 4.49 (s, 2H) ), 4.00 (q, J = 7.2 Hz, 2H), 3.78 (t, J = 7.2 Hz, 2H), 3.46 (s, 2H), 1.85-1.77 (m, 2H), 1.22 (t, J = 7.2 Hz , 3H), 1.03 (t, J = 7.2 Hz, 3H).LC / MS[M+H]395.2 (calculated value), LC / MS[M+H]395.1 (observed value).

[0191] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[5-[2-amino-4-[ethoxy(propyl)carbamoyl]- 3H-1-Benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-8a preparation of HxBz-5 (70 mg, 149 umol, 1.0 eq., 2HCl) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2- [3-Oxo-3-(2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (127mg, 179umol, 1.2eq) DIEA (77.4mg, 599umol, 104uL, 4.0eq) was added to a mixture of N 2 was added in one portion at 25° C. and the mixture was stirred at 25° C. for 0.5 h. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex luna C18 80*40mm*3um, mobile phase: [water (0.04%HCl)-ACN], B%: 12%-39%, 5.5 min). Purification provided HxBzL-8a (50.0 mg, 53.4 umol, 35.7% yield) as a yellow oil. 1 H NMR (400 MHz, MeOD) δ9.14 (s, 2H), 7.86-7.81 (m, 1H), 7.78-7.74 (m, 2H), 7.48 (s, 1H), 4.72 (s, 2H), 4.00 (q, J = 7.2 Hz, 2H), 3.85-3.71 (m, 8H), 3.69-3.58 (m, 38H), 3.47 (s, 2H), 2.62 (t, J = 6.0 Hz, 2H), 2.55 ( t, J = 6.4 Hz, 2H), 1.85-1.76 (m, 2H), 1.23 (t, J = 7.2 Hz, 3H), 1.03 (t, J = 7.2 Hz, 3H).

[0192] Preparation of HxBzL-8 HxBzL-8a (60 mg, 61.7 umol, 1.0 eq., HCl) and (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy sodium ( EDCI (71.0mg, 370umol, 6.0eq) was added to a mixture of N 2 was added in one portion at 25° C. and the mixture was stirred at 25° C. for 1 h. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1% TFA)-ACN], B%: 20%-45%, 8 min). Purification provided HxBzL-8 (38.0 mg, 30.5 umol, 49.3% yield, 93.3% purity) as a yellow oil. 1 H NMR (400 MHz, MeOD) δ9.11 (s, 2H), 7.83-7.79 (m, 1H), 7.77 (s, 1H), 7.76-7.71 (m, 1H), 7.47 (s, 1H), 4.71 (s, 2H), 4.00 (q, J = 7.2 Hz, 2H), 3.88 (t, J = 5.6 Hz, 2H), 3.85-3.75 (m, 5H), 3.70-3.57 (m, 38H), 3.47 ( s, 2H), 2.99 (t, J = 6.0 Hz, 2H), 2.62 (t, J = 4 Hz, 2H), 1.85-1.75 (m, 2H), 1.23 (t, J = 7.2 Hz, 3H), 1.02 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1163.3 (calculated value), LC / MS[M+H]1163.3 (observed value).

[0193] Example L-10 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[1-[[5-[2 -amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]-3-pyridyl]sulfonyl]azetidin-3-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-10 [ka] Preparation of tert-butyl ((1-((5-bromopyridin-3-yl)sulfonyl)azetidin-3-yl)methyl)carbamate, HxBz-7b tert-Butyl N-(azetidin-3-ylmethyl)carbamate (762 mg, 4.09 mmol, 1.05 eq) and 5-bromopyridine-3-sulfonyl chloride, HxBz-7a (1 g, 3.90 mmol, 2.26 mL) in DCM (20 mL) , 1 equivalent) in a mixture of Et 3 N (789 mg, 7.80 mmol, 1.09 mL, 2 equivalents), N 2 was added at 25° C. and then stirred at 25° C. for 1 hour. In the mixture, add H 2 O (20 mL) was added and then concentrated in vacuo to remove DCM. The desired solid precipitated from the mixture was filtered to give the desired product HxBz-7b (1.1 g, 2.71 mmol, 69.45% yield) as a white solid. 1 H NMR (DMSO-d 6 , 400MHz) δ9.09 (d, J = 2.0 Hz, 1H), 8.93 (d, J = 2.0 Hz, 1H), 8.40 (t, J = 2.0 Hz, 1H), 6.90 (t, J = 6.0 Hz, 1H), 3.80 (t, J = 8.4 Hz, 2H), 3.52 (dd, J = 6.0, 8.0 Hz, 2H), 2.93 (t, J = 6.0 Hz, 2H), 2.56-2.52 (m, 1H), 1.34 (s, 9H).

[0194] tert-Butyl((1-((5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)sulfonyl)azetidin-3-yl)methyl ) Preparation of carbamate, HxBz-7c HxBz-7b (0.75 g, 1.85 mmol, 1 eq) 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-) in dioxane (15 mL) Dioxaborolane-2-yl)-1,3,2-dioxaborolane, Pin 2 B 2 , bis(pinacolato)diboron, CAS Reg.No.78183-34-3 (703 mg, 2.77 mmol, 1.5 eq.) KOAc (362 mg, 3.69 mmol, 2 eq.) in a mixture of Pd(dppf)Cl 2 (67.5mg, 92.3umol, 0.05eq), N 2 The mixture was added at 25° C. and then stirred at 100° C. for 1 hour. The mixture was filtered and concentrated in vacuo. HxBz-7c (0.85 g, crude) was obtained as a yellow oil.

[0195] tert-Butyl((1-((5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyridin-3-yl)sulfonyl)azetidine-3- Preparation of yl)methyl)carbamate, HxBz-7d HxBz-7c (0.85 g, 1.87 mmol, 1 eq.) and 2-amino-8-bromo-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide, HxBzL-2b ( 755 mg, 2.06 mmol, 1.1 eq. 2 K in O (3 mL) 2 C.O. 3 (518mg, 3.75mmol, 2eq) and Pd(dppf)Cl 2 (68.6mg, 93.7umol, 0.05eq), N 2 The mixture was added at 25° C. and stirred at 100° C. for 1 hour. Mixture, H 2 Pour into O (50 mL). The aqueous phase was extracted with ethyl acetate (150 mL*3). The combined organic phases were washed with brine (100 mL) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100 to 200 mesh silica gel, petroleum ether / ethyl acetate = 5 / 1, 0 / 1 to EtOAc / MeOH = 10 / 1). HxBz-7d (1 g, 1.63 mmol, yield 87.05%) was obtained as an off-white solid. 1 H NMR (DMSO-d 6 , 400MHz) δ9.18 (d, J = 2.0 Hz, 1H), 8.95 (d, J = 2.0 Hz, 1H), 8.42 (t, J = 2.0 Hz, 1H), 7.55-7.51 (m, 2H), 7.49-7.45 (m, 1H), 7.30 (s, 1H), 3.96 (q, J = 7.6 Hz 2H), 3.90 (t, J = 8.0 Hz, 2H), 3.74 (t, J = 7.2 Hz, 2H) , 3.60 (dd, J = 6.0, 8.0 Hz, 2H), 3.35 (s, 2H),3.06 (d, J = 6.0 Hz, 2H), 2.69-2.58 (m, 1H), 1.77 (sxt, J = 7.2 Hz, 2H), 1.36 (s, 9H), 1.17 (t, J = 7.2 Hz, 3H), 0.99 (t, J = 7.2 Hz, 3H).

[0196] 2-Amino-8-[5-[3-(aminomethyl)azetidin-1-yl]sulfonyl-3-pyridyl]-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide, HxBz-7 preparation of CH 3 CN (10mL) and H 2 To a mixture of HxBz-7d (0.8g, 1.31mmol, 1eq) in O (10mL) was added TFA (1.49g, 13.1mmol, 967uL, 10eq) with N 2 The mixture was added at 25° C. and then stirred at 80° C. for 1 hour. The mixture was concentrated in vacuo to CH 3 CN was removed, the aqueous solution was extracted with MTBE (20*3) and discarded, and then the aqueous phase was directly lyophilized to produce HxBz-7 (0.9 g, 1.22 mmol, yield 93.07%, 2TFA) as off-white Obtained as a solid. 1 H NMR (MeOD, 400MHz) δ9.24 (d, J = 2.0 Hz, 1H), 9.04 (d, J = 2.0 Hz, 1H), 8.50 (t, J = 2.0 Hz, 1H), 7.87-7.78 (m , 2H), 7.77-7.72 (m, 1H), 7.46 (s, 1H), 4.06-3.94 (m, 4H), 3.79-3.70 (m, 4H), 3.45 (s, 2H), 3.12 (d, J = 7.6 Hz, 2H), 2.83-2.73 (m, 1H), 1.79 (sxt, J = 7.2 Hz, 2H), 1.20 (t, J = 7.2 Hz, 3H), 1.01 (t, J = 7.2 Hz, 3H) ).LC / MS[M+H]513.2 (calculated value), LC / MS[M+H]513.2 (observed value).

[0197] 1-(1-((5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyridin-3-yl)sulfonyl)azetidin-3-yl) Preparation of -3-oxo-6,9,12,15,18,21,24,27,30,33-decaoxa-2-azahexatriacontan-36-acid, HxBzL-10a Et 3 N (161mg, 1.60mmol, 222uL, 2.5eq), N 2 was added at 0° C. and then stirred at 0° C. for 1 hour. H the mixture 2 The pH of the mixture was adjusted to ~6 with TFA at 0 °C, then extracted with MTBE (10 mL) and discarded, and the aqueous phase was further extracted with DCM / i-PrOH (20 mL*3). did. Mix organic phase with anhydrous Na 2 S.O. 4 , filtered and concentrated in vacuo to give HxBzL-10a (0.6 g, 569.68 umol, 89.25% yield) as a pale yellow oil.

[0198] Preparation of HxBzL-10 HxBzL-10a (0.6g, 570umol, 1 eq) and (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxysodium (611mg, 2.28g) in DCM (10mL) and DMA (1.5mL) EDCI (437 mg, 2.28 mmol, 4 eq.) was added to a mixture of N 2 was added at 25° C. and then stirred at 25° C. for 0.5 h. The mixture was concentrated in vacuo. The residue was filtered and purified by preparative HPLC column: Phenomenex luna C18 250*50mm*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 30%~50%, 10 minutes). HxBzL-10 (370 mg, 288.76 umol, yield 50.69%) was obtained as a white solid. 1 H NMR (MeOD, 400MHz) δ9.24 (d, J = 2.0 Hz, 1H), 9.03 (d, J = 2.0 Hz, 1H), 8.51 (t, J = 2.0 Hz, 1H), 7.91-7.84 (m , 2H), 7.74 (d, J = 8.8 Hz, 1H), 7.47 (s, 1H), 4.03-3.91 (m, 4H), 3.86 (t, J = 6.0 Hz, 2H), 3.76 (t, J = 7.2 Hz, 2H), 3.66-3.49 (m, 40H), 3.47 (s, 2H), 3.21 (d, J = 6.4 Hz, 2H), 3.01-2.92 (m, 2H), 2.79-2.68 (m, 1H) ), 2.29 (t, J = 6.0 Hz, 2H), 1.78 (sxt, J = 7.2 Hz, 2H), 1.21 (t, J = 7.2 Hz, 3H), 1.01 (t, J = 7.2 Hz, 3H). LC / MS[M+H] 1281.5 (calculated value), LC / MS[M+H] 1281.6 (observed value).

[0199] Example L-15 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[2-(cyclobutoxy-carbonylamino)ethoxy-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-15 [ka] Cyclobutyl N-[2-[[2-amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carbonyl]-propyl-amino]oxyethyl ] Preparation of carbamate, HxBz-15b 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBz- in DCM (4 mL) and DMA (2 mL). EDCI (468 mg, 2.44 mmol, 4.0 eq.) was added to a mixture of 15a (250 mg, 611 umol, 1.0 eq.) and cyclobutyl N-[2-(propylaminooxy)ethyl]carbamate (201 mg, 794 umol, 1.3 eq., HCl). N 2 The mixture was added in one portion at 25° C. and stirred at 25° C. for 2 hours. DCM (4 mL) was removed in vacuo, water (10 mL) was added, the aqueous phase was extracted with ethyl acetate (10 mL*3), the combined organic phases were washed with brine (5 mL*2), anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100~200 mesh silica gel, petroleum ether / ethyl acetate = 10 / 1, 0 / 1 ~ ethyl acetate / methanol = 10 / 1) to obtain HxBz. -15b (190 mg, 313 umol, 51.2% yield) was obtained as a brown oil. 1 H NMR (400 MHz, MeOD) δ9.08 (s, 2H), 7.63 (d, J = 8.0 Hz, 1H), 7.58-7.52 (m, 2H), 7.37 (s, 1H), 4.74-4.67 (m , 2H), 4.54 (s, 2H), 3.96 (t, J = 4.8 Hz, 2H), 3.76 (t, J = 7.2 Hz, 2H), 3.33 (s, 2H), 2.20 (dd, J = 2.8, 5.2 Hz, 2H), 1.94-1.86 (m, 2H), 1.82-1.75 (m, 2H), 1.50 (s, 9H), 1.38 (d, J = 1.6 Hz, 2H), 1.01 (t, J = 7.2 Hz, 3H).

[0200] Cyclobutyl N-[2-[[2-amino-8-[2-(aminomethyl)pyrimidin-5-yl]-3H-1-benzazepine-4-carbonyl]-propyl-amino]oxyethyl]carbamate, HxBz-15 preparation of CF 3 COOH (535mg, 4.69mmol, 347uL, 15eq) was converted into N 2 was added in one portion at 25° C. and then stirred at 25° C. for 1.5 hours. DCM (5 mL) was removed in vacuo, the residue was diluted with water (10 mL), the aqueous phase was extracted with MTBE (5 mL*4) to remove excess TFA, and then the aqueous phase was lyophilized. , HxBz-15 (130 mg, 169 umol, yield 54.1%, purity 95.7%, 2TFA) was obtained as a brown solid. 1 H NMR (400 MHz, MeOD) δ = 9.21 (s, 2H), 7.85-7.76 (m, 3H), 7.49 (s, 1H), 4.66 (t, J = 7.2 Hz, 1H), 4.48 (s, 2H) ), 3.96 (t, J = 5.2 Hz, 2H), 3.76 (t, J = 7.2 Hz, 2H), 3.43 (s, 2H), 3.31 (s, 2H), 2.20-2.10 (m, 2H), 1.91 -1.83 (m, 2H), 1.81-1.74 (m, 2H), 1.70-1.60 (m, 1H), 1.57-1.47 (m, 1H), 1.00 (t, J = 7.2 Hz, 3H).LC / MS [M+H]508.3 (calculated value), LC / MS [M+H]508.1 (observed value).

[0201] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[2-(cyclobutoxycarbonyl amino)ethoxy-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]ethoxy]propanoic acid, preparation of HxBzL-15a HxBz-15 (105 mg, 181 umol, 1.0 eq., 2HCl) and Et in DMF (1.5 mL) 3 A mixture of 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-oxo- 3-(2,3,4,5,6-pentafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, PFP-PEG10-CO2H (131mg, 181umol, 1.0 equivalent), N 2 The mixture was added at 0° C. and stirred at 0° C. for 0.5 h, then at 25° C. for an additional 0.5 h. The reaction mixture was concentrated, the residue was diluted with water (5 mL), the aqueous phase was extracted with ethyl acetate (3 mL*2) and discarded, then the aqueous phase was extracted with DCM / iPrOH=3 / 1 (5 mL*3) The mixed organic layer was further extracted with anhydrous Na 2 S.O. 4 , filtered and concentrated in vacuo to give HxBzL-15a (100 mg, 95.4 umol, 52.7% yield) as a yellow oil.

[0202] Preparation of HxBzL-15 HxBzL-15a (100 mg, 95.4 umol, 1.0 eq.) and sodium (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy (128 mg, 477 umol) in DCM (1 mL) and DMA (0.5 mL). , 5.0 eq.), EDCI (91.4 mg, 477 umol, 5.0 eq.) was added to a mixture of N 2 was added in one portion at 25° C. and then stirred at 25° C. for 1 hour. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 15%~35%, 8 min). Purification provided HxBzL-15 (35.1 mg, 25.6 umol, 26.9% yield, 93.3% purity) as a pale yellow oil.1 H NMR (400 MHz, MeOD) δ9.12 (s, 2H), 7.84-7.77 (m, 3H), 7.52 (s, 1H), 4.75-4.67 (m, 3H), 3.99 (t, J = 5.2 Hz , 2H), 3.88 (t, J = 6.0 Hz, 2H), 3.82 (t, J = 6.0 Hz, 2H), 3.78 (t, J = 7.2 Hz, 2H), 3.70-3.57 (m, 38H), 3.45 (s, 2H), 3.01-2.97 (m, 2H), 2.62 (t, J = 6.0 Hz, 2H), 2.24-2.14 (m, 2H), 1.96-1.86 (m, 2H), 1.84-1.75 (m , 2H), 1.73-1.61 (m, 1H), 1.59-1.49 (m, 1H), 1.01 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1276.5 (calculated value), LC / MS[M+H]1276.6 (observed value).

[0203] Example L-16 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[2-(Cyclobutylcarbamoylamino)ethoxy-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy Synthesis of ]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBZL-16 [ka] tert-Butyl((5-(2-amino-4-((2-(3-cyclobutylureido)ethoxy)(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyrimidin-2-yl )Methyl)carbamate, preparation of HxBz-16a 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBz- in DCM (2 mL) and DMA (2 mL). 15a (250 mg, 611 umol, 1 eq.) in a solution of 1-cyclobutyl-3-[2-(propylaminooxy)ethyl]urea (231 mg, 916 umol, 1.5 eq., HCl) with EDCI (351 mg, 1.83 mmol, 3 eq.) was added and stirred at 25°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to remove DCM. The residue was diluted with water (10 mL) and extracted with EtOAc (20 mL*3). Wash the mixed organic layer with brine (20mL*2) and remove Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate=50 / 1 to ethyl acetate:MeOH=5:1) to obtain HxBz-16a (230 mg, 380 umol, yield 62.1%) as a brown solid.

[0204] 2-Amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-[2-(cyclobutylcarbamoylamino)ethoxy]-N-propyl-3H-1-benzazepine-4-carboxamide, HxBz- Preparation of 16 To a solution of HxBz-16a (230 mg, 0.38 mmol, 1 eq) in water (2 mL) and MeCN (2 mL) was added TFA (432 mg, 3.79 mmol, 0.28 mL, 10 eq), then at 80 °C for 0.5 h. Stirred. The mixture was concentrated under reduced pressure, the residue was diluted with water (2 mL), extracted with MTBE (3 mL*3) and discarded, and the aqueous phase was concentrated under reduced pressure to obtain HxBz-16 (230 mg, 371 umol, yield 97.8% TFA) was obtained as a brown solid. 1 H NMR (400 MHz, MeOD) δ 9.21 (s, 2H), 7.84-7.73 (m, 3H), 7.47 (s, 1H), 4.48 (s, 2H), 4.01-3.89 (m, 3H), 3.75 ( t, J = 7.2 Hz, 2H), 3.44 (s, 2H), 3.33 (br s, 2H), 2.19-2.10 (m, 2H), 1.81-1.68 (m, 4H), 1.64-1.55 (m, 2H) ), 1.00 (t, J = 7.2 Hz, 3H).LC / MS[M+H]507.3 (calculated value), LC / MS[M+H]507.2 (observed value).

[0205] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[2-(cyclobutylcarbamoyl amino)ethoxy-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]Ethoxy]propanoic acid, preparation of HxBzL-16a HxBz-16 (100mg, 136umol, 1eq, 2TFA) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[ 3-Oxo-3-(2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (96.2 mg, 0.14 mmol, 1 eq. ) in a solution of Et 3 N (41.3 mg, 0.41 mmol, 56.8 uL, 3 eq.) was added and then stirred at 25° C. for 0.5 h. The pH of the mixture was adjusted to ~6 with TFA at 0 °C, extracted with EtOAc (5 mL, 3 times) and discarded, and the aqueous solution was further extracted with DCM / i-PrOH (10 mL*3, 3 / 1). did. Organic layer, Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure. The crude product HxBzL-16a (120 mg, 115 umol, 84.2% yield) was obtained as a yellow oil and used in the next step without further purification.

[0206] Preparation of HxBzL-16 HxBzL-16a (70 mg, 66.9 umol, 1 eq.) and sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (71.7 mg, 267 umol) in DMA (0.5 mL) and DCM (1.5 mL). , 4 eq.) was added EDCI (51.3 mg, 267 umol, 4 eq.) and stirred at 25° C. for 0.5 h. The mixture was filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 15% to 35%, 8 minutes). . The residue was then purified by preparative HPLC (TFA conditions, column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 15%~35%, 8 minutes). Purification provided HxBzL-13 (20 mg, 13.3 umol, 19.9% ​​yield, 2TFA) as a colorless oil. 1 H NMR (400 MHz, MeOD) δ 9.09 (s, 2H), 7.80-7.71 (m, 3H), 7.47 (s, 1H), 4.69 (s, 2H), 3.95 (br t, J = 5.2 Hz, 2H ), 3.86 (t, J = 6.0 Hz, 2H), 3.80 (t, J = 6.0 Hz, 2H), 3.75 (br t, J = 7.2 Hz, 2H), 3.68-3.57 (m, 38H), 3.45 ( s, 2H), 2.97 (t, J = 6.0 Hz, 2H), 2.60 (t, J = 6.0 Hz, 2H), 2.15 (br d, J = 7.2 Hz, 2H), 1.83-1.68 (m, 4H) , 1.64-1.52 (m, 2H), 0.99 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1275.5 (calculated value), LC / MS[M+H]1275.2 (observed value).

[0207] Example L-17 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[3-(Cyclobutoxycarbonylamino)propyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy Synthesis of ]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBZL-17 [ka] Cyclobutyl N-[3-[[2-amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carbonyl]-propyl-amino]propyl ], Preparation of HxBz-14 2-amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBz-14a (0.25 g, Et 3 N (185 mg, 1.83 mmol, 255 uL, 3.0 eq.), cyclobutyl N-[3-(propylamino)propyl]carbamate (170 mg, 678 umol, 1.11 eq., HCl), and azabenzotriazole tetramethyluronium hexafluorophosphate, HATU (232 mg, 611 umol, 1.0 eq) was added in one portion at 0°C and stirred at 0°C for 0.5 h. The mixture was then diluted with water and extracted with EtOAc (20 mL x 3). Wash the organic layer with brine and remove Na 2 S.O. 4Dry on top, filter and concentrate. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 3 / 1) to obtain HxBz-14 (0.28 g, 462 umol, yield). 75.71%) as a yellow solid. 1 H NMR (MeOD, 400 MHz) δ9.04 (s, 2H), 7.52 (d, J = 8.4 Hz, 1H), 7.48 (d, J = 1.6 Hz, 1H), 7.45-7.40 (m, 1H), 6.93 (s, 1H), 4.84-4.84 (m, 1H), 4.64 (s, 4H), 3.54-3.47 (m, 2H), 3.46-3.39 (m, 2H), 3.30 (m, 2H), 3.22- 3.07 (m, 2H), 2.32-2.28 (m, 2H), 2.10-2.00 (m, 2H), 1.88-1.79 (m, 3H), 1.75-1.60 (m, 3H), 1.48 (s, 9H), 0.90 (s, 3H).LC / MS[M+H]606.3 (calculated value), LC / MS[M+H]606.2 (observed value).

[0208] Cyclobutyl N-[3-[[2-amino-8-[2-(aminomethyl)pyrimidin-5-yl]-3H-1-benzazepine-4-carbonyl]-propyl-amino]propyl]carbamate, HxBz-13 preparation of CH 3 CN (3mL) and H 2 To a mixture of HxBz-14 (0.26 g, 429 umol, 1.0 eq.) in O (1 mL) was added TFA (489 mg, 4.29 mmol, 318 uL, 10.0 eq.) in one portion at 25 °C, then stirred at 80 °C for 0.5 h. did. The mixture was then concentrated, the residue was diluted with water (10 mL), and the mixture was extracted with MTBE (10 mL x 2) to remove excess TFA. The aqueous layer was freeze-dried to obtain HxBz-13 (0.2 g, 323 umol, yield 75.20%, TFA) as a yellow solid. 1 H NMR (MeOD, 400 MHz) δ9.21 (s, 2H), 7.84-7.71 (m, 3H), 7.12 (s, 1H), 4.85-4.85 (m, 1H), 4.47 (s, 2H), 3.54 (t, J = 7.2 Hz, 2H), 3.48 (s, 2H), 3.37 (s, 2H), 3.15 (d, J = 15.6 Hz, 2H), 2.30-2.25 (m, 2H), 2.08-2.00 ( m, 2H), 1.89-1.66 (m, 6H), 1.01-0.88 (m, 3H).LC / MS[M+H]506.3 (calculated value), LC / MS[M+H]506.2 (observed value) .

[0209] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[3-(cyclobutoxycarbonyl amino)propyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]ethoxy]propanoic acid, preparation of HxBzL-17a Et 3 N (48.9mg, 484umol, 67.4uL, 3.0eq) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-oxo-3- (2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, TFP-PEG10-CO2H (114 mg, 161 umol, 1.0 equiv) , added in one portion at 0°C and then stirred at 0°C for 0.5 h. The pH of the mixture was adjusted to 5-6 with TFA at 0 °C. The mixture was then diluted with water (5 mL) and washed with MTBE (10 mL x 3). The aqueous layer was then further extracted with DCM:i-PrOH=3:1 (20 mL x 3). organic layer with Na 2 Dry over SO4, filter, and concentrate to give HxBzL-17a (0.15 g, 129 umol, 80.11% yield, TFA) as a yellow oil.

[0210] Preparation of HxBzL-17 Sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate ( 139 mg, 517 umol, 4.0 eq) and EDCI (149 mg, 776 umol, 6.0 eq) were added in one portion at 25°C and then stirred at 25°C for 0.5 h. The mixture was concentrated and filtered. The residue was then purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1% TFA)-ACN], B%: 15%~40%, 8 min). , HxBzL-17 (75.3 mg, 59.1 umol, yield 45.71%) was obtained as a yellow oil. 1 H NMR (MeOD, 400 MHz) δ9.09 (s, 2H), 7.82-7.67 (m, 3H), 7.11 (s, 1H), 4.86-4.82 (m, 1H), 4.69 (s, 2H), 3.86 (t, J = 6.0 Hz, 2H), 3.80 (t, J = 6.0 Hz, 2H), 3.66-3.48 (m, 40H), 3.38 (s, 2H), 3.22-3.06 (m, 2H), 2.97 ( t, J = 6.0 Hz, 2H), 2.64-2.58(m, 2H), 2.32-2.25 (m, 2H), 2.09-1.95 (m, 2H), 1.91-1.80 (m, 3H), 1.75-1.61 ( m, 3H), 0.93 (s, 3H).LC / MS[M+H]1274.5 (calculated value), LC / MS[M+H]1274.3 (observed value).

[0211] Example L-18 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[5-[2-amino-4 -[Ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy ]-2,3,5,6-tetrafluoro-benzenesulfonic acid, synthesis of HxBzL-18 [ka] Preparation of 2-amino-8-bromo-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide, HxBz-11b N-ethoxypropan-1-amine (9.6 g, 68.8 mmol, 1.3 eq., HCl) and 2-amino-8-bromo-3H-1-benzazepine-4-carboxylic acid in DMA (150 mL) and DCM (150 mL). , EDCI (40.6 g, 211 mmol, 4.0 eq.) was added to a mixture of HxBz-11a (14.8 g, 52.9 mmol, 1.0 eq.) with N 2 The mixture was added at 25°C. The mixture was stirred at 25°C for 2 hours. Adjust the pH of the mixture to NaHCO 3 9 and concentrated under reduced pressure to remove DCM at 45°C. The aqueous phase was extracted with ethyl acetate (100 mL x 3). The combined organic layer was washed with brine (1000mLx2) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was triturated with MTBE / PE=1 / 1 at 25° C. to obtain HxBz-11b (12.5 g, 34.1 mmol, yield 64.5%) as a white solid. 1 H NMR (MeOD, 400MHz) δ7.31 (d, J = 2.0 Hz, 1H), 7.26-7.22 (m, 1H), 7.18 (s, 1H), 7.17-7.14 (m, 1H), 3.92 (q, J = 6.8 Hz, 2H), 3.71 (t, J = 7.2 Hz, 2H), 3.31 (s, 2H), 1.79-1.70 (m, 2H), 1.15 (t, J = 7.2 Hz, 3H), 0.97 ( t, J = 7.6 Hz, 3H).

[0212] 2-Amino-N-ethoxy-N-propyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3H-1-benzazepine-4-carboxamide, HxBz Preparation of -11c HxBz-11b (500 mg, 1.37 mmol, 1.0 equiv.) in dioxane (10 mL), Pin 2 B 2 (416mg, 1.64mmol, 1.2eq), KOAc (335mg, 3.41mmol, 2.5eq), and Pd(dppf)Cl 2 (99.9mg, 136umol, 0.1eq) was degassed and N 2 Purge the mixture three times with N 2 Stirred at 95° C. for 1 hour under atmosphere. The mixture was concentrated in vacuo. The residue was poured into ice water (w / w=1 / 1) (10 mL) and stirred for 5 minutes. The aqueous phase was extracted with MTBE (10 mL x 1), then the aqueous phase was further extracted with DCM / i-PrOH=3 / 1 (10 mL x 3). The mixed organic layer (DCM / i-PrOH) was diluted with anhydrous Na 2 S.O. 4 , filtered and concentrated in vacuo to give HxBz-11c (490 mg, crude), which was used in the next step without further purification as a black solid.

[0213] Preparation of methyl 5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyrimidine-2-carboxylate, HxBz-11 Dioxane (15mL) and H 2 HxBz-11c (390 mg, 944 umol, 1.0 eq.), methyl 5-bromopyrimidine-2-carboxylate (266 mg, 1.23 mmol, 1.3 eq.), Pd(dppf)Cl in O (2 mL). 2 (69.0mg, 94.3umol, 0.1eq), K 3 P.O. 4 (401 mg, 1.89 mmol, 2.0 eq.) was degassed and N 2 Purge 3 times with N 2 Stirred at 80°C for 1 hour under atmosphere. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 5%~30%, 8 min) to obtain HxBz -11 (105 mg, 161 umol, yield 17.1%, TFA) was obtained as a white solid. 1 H NMR (MeOD, 400MHz) δ9.30 (s, 2H), 7.89 (dd, J = 2.0, 2.0 Hz, 1H), 7.83-7.74 (m, 2H), 7.47 (s, 1H), 4.06 (s, 3H), 4.00 (t, J = 6.8 Hz, 2H), 3.76 (t, J = 7.2 Hz, 2H), 3.45 (s, 2H), 1.83-1.74 (m, 2H), 1.21 (t, J = 6.8 Hz, 3H), 1.01 (t, J = 7.2 Hz, 3H).LC / MS[M+H]424.1 (calculated value), LC / MS[M+H]424.1 (observed value).

[0214] Preparation of 5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidine-2-carboxylic acid, HxBzL-18a EtOH (5 mL) and H 2 A solution of HxBz-11 (330 mg, 779 umol, 1.0 eq.) in O (0.5 mL) was added with 2 O (131 mg, 3.12 mmol, 4.0 eq.) was added. The mixture was stirred at 25°C for 2 hours. The pH of the mixture was adjusted to ~6 with HCl (4M) and concentrated in vacuo to remove EtOH. The residue was diluted with water (10 mL). The aqueous phase was extracted with DCM / i-PrOH=3 / 1 (10 mL x 3). Mix organic layer with anhydrous Na 2 S.O. 4 , filtered and concentrated in vacuo to give HxBzL-18a (200 mg, 488 umol, 62.7% yield) as a yellow solid.

[0215] Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[[1-[[5-[2-amino-4- [3-(3,3-dimethylbutanoylamino)propyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]-3-pyridyl]sulfonyl]azetidin-3-yl]methyl-methyl-amino]ethoxy Preparation of ]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate, HxBzL-18b HxBzL-18a (195 mg, 332 umol, 0.8 eq.) and tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2- (2-Aminoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate, tBuOOC-PEG 10 -NH 2 (390mg, 666umol, 1.0eq.) to a mixture of Et 3 N (126 mg, 1.25 mmol, 173 uL, 3.0 eq) and HATU (158 mg, 415 umol, 1.0 eq) were added at 0°C. The mixture was stirred at 0°C for 1 hour. The mixture was purified by preparative HPLC (column: Phenomenex luna C18 80*40mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 25%~50%, 7 min) to obtain HxBzL. -18b (80 mg, 66.4 umol, 16.0% yield, TFA) was obtained as a yellow oil.

[0216] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl] -3H-1-Benzazepin-8-yl]pyrimidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, preparation of HxBzL-18c To a solution of HxBzL-18b (80 mg, 66.4 umol, 1.0 eq., TFA) in MeCN (2 mL) and H2O (1 mL) was added HCl (12M, 83.0 uL, 15.0 eq.) and stirred at 80 °C for 1 h. . The mixture was concentrated in vacuo to give a residue, which was lyophilized to give HxBzL-18c (60 mg, 62.7 umol, 94.4% yield, HCl) as a colorless oil.

[0217] Preparation of HxBzL-18 HxBzL-18c (60 mg, 60.4 umol, 1.0 eq., 2HCl) and (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy sodium (64.7 EDCI (46.3 mg, 241 umol, 4.0 eq.) was added to a solution of EDCI (46.3 mg, 241 umol, 4.0 eq.) and then stirred at 25.degree. The mixture was concentrated in vacuo and filtered. The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1% TFA)-ACN], B%: 15%~35%, 8 min) to obtain HxBzL. -18 (36 mg, 31.3 umol, 51.9% yield) was obtained as a yellow oil. 1 H NMR (MeOD, 400MHz) δ9.27 (s, 2H), 7.90-7.81 (m, 2H), 7.75 (d, J = 8.4 Hz, 1H), 7.46 (s, 1H), 3.98 (q, J = 6.8 Hz, 2H), 3.85 (t, J = 6.0 Hz, 2H), 3.78-3.75 (m, 2H), 3.73-3.72 (m, 2H), 3.70-3.56 (m, 36H), 3.46 (s, 2H) ), 2.96 (t, J = 6.0 Hz, 2H), 1.84-1.71 (m, 2H), 1.21 (t, J = 6.8 Hz, 3H), 1.00 (t, J = 7.6 Hz, 3H).LC / MS [M+H] 1149.4 (calculated value), LC / MS [M+H] 1149.5 (observed value).

[0218] Example L-19 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2S)-1-[5 -[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-19 [ka] Preparation of methyl (2S)-1-(5-bromopyrimidine-2-carbonyl)pyrrolidine-2-carboxyl-ate, HxBzL-19b 5-bromopyrimidine-2-carboxylic acid, HxBzL-19a (400mg, 1.97mmol, 1.0eq) in DMF (8mL), Et 3 HATU (749 mg, 1.97 mmol, 1.0 eq.) was added to a mixture of N (598 mg, 5.91 mmol, 822 uL, 3.0 eq.) and methyl (2S)-pyrrolidine-2-carboxylate (342 mg, 2.07 mmol, 1.05 eq., HCl). ,N 2 Added in one portion at 0° C. and stirred at 0° C. for 30 min, then heated to 25° C. and stirred for an additional 0.5 h. Water (20 mL) was added, the aqueous phase was extracted with ethyl acetate (20 mL*4), the combined organic phase was washed with brine (10 mL*1), anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 10 / 1, 4 / 1) to obtain HxBzL-19b (320 mg, 1.02 mmol, yield). 51.7%) was obtained as a yellow oil.

[0219] Preparation of (2S)-1-(5-bromopyrimidine-2-carbonyl)pyrrolidine-2-carboxylic acid, HxBzL-19c MeOH (5 mL) and H 2 A solution of HxBzL-19b (320 mg, 1.02 mmol, 1.0 equiv) in O (5 mL) was added with LiOH H 2 O (171mg, 4.07mmol, 4.0eq), N 2 The mixture was added in one portion at 25° C. and stirred at 25° C. for 2 hours. The reaction mixture was quenched with HCl (4 M) until pH=7, MeOH (5 mL) was removed in vacuo, and the desired solid precipitated from the aqueous phase, filtered, dried and purified with HxBzL-19c (300 mg, crude The product was obtained as a pale yellow solid.

[0220] Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2S)-1-(5-bromopyrimidine-2 -Carbonyl)pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanate, preparation of HxBzL-19d HxBzL-19c (200mg, 666umol, 1.0eq) in DMF (1mL), Et 3 N (168mg, 1.67mmol, 232uL, 2.5eq) and tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-aminoethoxy )ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanate (390mg, 666umol, 1.0eq) was added with N 2 Added in one portion at 0° C. and stirred at 0° C. for 30 min, then heated to 25° C. and stirred for an additional 0.5 h. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex luna C18 250*50mm*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 20% to 60%, 10 minutes). Purification provided HxBzL-19d (300 mg, 346 umol, 51.8% yield) as a colorless oil.

[0221] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2S)-1-(5-bromopyrimidine-2-carbonyl) Preparation of pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-19e MeCN (1 mL) and H 2 To a solution of HxBzL-19d (300 mg, 345 umol, 1.0 eq.) in O (3 mL) was added HCl (12 M, 864 uL, 30 eq.) with N 2 was added in one portion at 25° C. and then stirred at 80° C. for 1 hour. The reaction mixture was concentrated in vacuo to give HxBzL-19e (250 mg, 307.99 umol, 89.09% yield) as a yellow oil.

[0222] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2S)-1-[5-[2-amino-4- [ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy Preparation of ethoxy]propanoic acid, HxBzL-19f Dioxane (3mL) and H 2 HxBzL-19e (150 mg, 185 umol, 1.0 equiv.) in O (0.3 mL), 2-amino-N-ethoxy-N-propyl-8-(4,4,5,5-tetramethyl-1,3,2- Dioxaborolan-2-yl)-3H-1-benzazepine-4-carboxamide (91.6mg, 222umol, 1.2eq), Pd(dppf)Cl 2 (13.5 mg, 18.5 umol, 0.1 equivalent) and K 2 C.O. 3 A solution of (63.8 mg, 462 umol, 2.5 eq.) was degassed and then N 2 The mixture was heated to 95°C for 2 hours under vacuum. The reaction mixture was filtered, the filtrate was concentrated in vacuo, and the residue was purified by preparative HPLC (column: Phenomenex luna C18 80*40mm*3um, mobile phase: [water (0.04%HCl)-ACN], B%: 5% ~45%, 7 min) to give HxBzL-19f (110 mg, 108 umol, 58.4% yield) as a yellow oil.

[0223] Preparation of HxBzL-19 HxBzL-19f (110 mg, 108 umol, 1.0 eq) and (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy sodium (145 mg, 540 umol, in DCM (2 mL) and DMA (0.5 mL)) EDCI (103mg, 540umol, 5.0eq) was added to a mixture of N 2 was added in one portion at 25° C. and stirred at 25° C. for 1 hour. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 10%~40%, 8 min). Purification provided HxBzL-19 (66.5 mg, 50.9 umol, 47.1% yield, 95.3% purity) as a pale yellow oil. 1 H NMR (400 MHz, MeOD) δ9.28-9.24 (m, 2H), 7.91-7.81 (m, 2H), 7.80-7.74 (m, 1H), 7.50-7.47 (m, 1H), 4.00 (q, J = 7.2 Hz, 2H), 3.88 (dt, J = 3.2, 5.6 Hz, 4H), 3.81-3.74 (m, 4H), 3.70-3.53 (m, 37H), 3.50-3.32 (m, 5H), 3.02 -2.96 (m, 2H), 2.16-1.97 (m, 4H), 1.84-1.76 (m, 2H), 1.23 (t, 7.2 Hz, 3H), 1.03 (t, 7.2 Hz, 3H).LC / MS[ M+H]1246.5 (calculated value), LC / MS[M+H]1246.7 (observed value).

[0224] Example L-24 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[2-(dimethylcarbamoylamino)ethoxy-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-24 [ka] tert-Butyl((5-(2-amino-4-((2-(3,3-dimethylureido)ethoxy)(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyrimidine-2- Preparation of yl)methyl)carbamate, HxBz-20a 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBz- in DCM (3 mL) and DMA (1 mL). EDCI (468 mg, 2.44 mmol) was added to a mixture of 14a (250 mg, 611 umol, 1 eq.) and 1,1-dimethyl-3-[2-(propylaminooxy)ethyl]urea (165 mg, 733 umol, 1.2 eq., HCl). , 4 equivalents) and stirred at 25°C for 1 hour. The mixture was concentrated in vacuo to remove DCM, the residue was diluted with water (10 mL), and the pH of the mixture was adjusted to Na 2 C.O. 3 It was adjusted to about 8 with an aqueous solution. The aqueous phase was extracted with ethyl acetate (10 mL*4). Wash the mixed organic layer with brine (20mL*1) and add anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. Purify the residue by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 0 / 1, ethyl acetate / methanol = 1 / 0, 3 / 1). HxBz-20a (260 mg, 447.75 umol, yield 73.33%) was obtained as a yellow solid.

[0225] Preparation of HxBz-20 To a solution of HxBz-20a (130 mg, 224 umol, 1 eq.) in EtOAc (3.00 mL) was added HCl / EtOAc (4M, 3.00 mL, 53.60 eq.) and then stirred at 25° C. for 1 h. The mixture was concentrated to give HxBz-20 (115 mg, 207.77 umol, 92.81% yield, 2HCl) as a pale red solid. 1 H NMR (MeOD, 400 MHz) δ9.22 (s, 2H), 7.86-7.80 (m, 2H), 7.80-7.74 (m, 1H), 7.50 (s, 1H), 4.48 (s, 2H), 3.97 (t, J = 5.2 Hz, 2H), 3.76 (t, J = 7.2 Hz, 2H), 3.45 (s, 2H), 3.38-3.34 (m, 2H), 2.74 (s, 6H), 1.83-1.73 ( m, 2H), 1.00 (t, J = 7.6 Hz, 3H).LC / MS[M+H]481.3 (calculated value), LC / MS[M+H]481.1 (observed value).

[0226] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[2-(dimethylcarbamoylamino )ethoxy-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Preparation of ethoxy]propanoic acid, HxBzL-24a A solution of HxBz-20 (65.0 mg, 117 umol, 1 eq., 2HCl) in DMF (1.00 mL) was added with Et 3N (48.0 mg, 470 umol, 4 eq.) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-oxo-3-(2, 3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-21a (83.0 mg, 117 umol, 1 eq.) was added, then Stirred at 0°C for 1 hour. The mixture was diluted with water (10 mL), the pH of the mixture was adjusted to about 6 by gradual addition of TFA, extracted with MTBE (10 mL) and discarded, and the aqueous solution was diluted with DCM:i-PrOH= It was further extracted with 3:1 (20 mL x 3). organic layer with Na 2 S.O. 4 Dry on top, filter and concentrate to give HxBzL-24a (95 mg, 93.03 umol, 79.22% yield) as a pale yellow oil.

[0227] Preparation of HxBzL-24 HxBzL-24a (90.0 mg, 88.1 umol, 1 eq) and (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy sodium (95.0 EDCI (68.0 mg, 353 umol, 4 eq.) was added to a solution of EDCI (68.0 mg, 353 umol, 4 eq.) and stirred at 25° C. for 1 hour. The mixture was concentrated and filtered. The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1% TFA)-ACN], B%: 5%~35%, 8 min) to obtain HxBzL. -24 (51 mg, 37.41 umol, 42.45% yield, TFA) was obtained as a pale yellow oil. 1 H NMR (MeOD, 400 MHz) δ9.10 (s, 2H), 7.83-7.70 (m, 3H), 7.48 (s, 1H), 4.69 (s, 2H), 3.97 (t, J = 5.2 Hz, 2H ), 3.86 (t, J = 5.6 Hz, 2H), 3.80 (t, J = 6.0 Hz, 2H), 3.78-3.74 (m, 2H), 3.65-3.55 (m, 36H), 3.45 (s, 2H) , 3.37-3.34 (m, 2H), 2.97 (t, J = 5.6 Hz, 2H), 2.74 (s, 6H), 2.60 (t, J = 6.0 Hz, 2H), 1.83-1.72 (m, 1H), 1.00 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1249.5 (calculated value), LC / MS[M+H]1249.6 (observed value).

[0228] Example L-26 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[2-hydroxyethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-26 [ka] Tert-Butyl N-[[5-[2-amino-4-[2-hydroxyethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methyl]carbamate, HxBz-22a preparation of 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBz in DCM (6 mL) and DMA (0.5 mL) EDCI (492 mg, 2.56 mmol, 3.0 eq.) was added once to a mixture of -14a (0.35 g, 855 umol, 1.0 eq.) and 2-(propylaminooxy)ethanol (200 mg, 1.28 mmol, 1.5 eq., HCl) at 25 °C. and then stirred at 25°C for 0.5 hour. The mixture was concentrated to remove DCM and the residue was purified with H 2 Diluted with O (10 mL). Adjust the pH of the mixture to NaHCO 3 It was adjusted to about 8 with an aqueous solution. The aqueous phase was then extracted with EtOAc (20 mL x 3). The organic layer was brine, Na 2 S.O. 4 Dry on top, filter and concentrate. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, ethyl acetate / MeOH=1 / 0, 10 / 1) to obtain HxBz-22a (0.37 g, 725 μmol, yield 84.77%) was obtained as a yellow oil. 1 H NMR (MeOD, 400 MHz) δ9.08-9.01 (m, 2H), 7.59 (d, J = 8.0 Hz, 1H), 7.54-7.46 (m, 2H), 7.40 (s, 1H), 4.56-4.49 (m, 2H), 4.02-3.95 (m, 2H), 3.81-3.74 (m, 2H), 3.73-3.66 (m, 2H), 1.88-1.72 (m, 2H), 1.48 (s, 9H), 0.99 (t, J = 7.6 Hz, 3H).

[0229] Preparation of 2-amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-(2-hydroxyethoxy)-N-propyl-3H-1-benzazepine-4-carboxamide, HxBz-22 H 2 O (4 mL) and CH 3 To a mixture of HxBz-22a (0.35 g, 685 umol, 1.0 eq.) in CN (0.5 mL) was added TFA (1.17 g, 10.3 mmol, 761 uL, 15.0 eq.) in one portion at 25 °C and then at 80 °C. Stirred for 0.5 hour. The mixture was extracted with MTBE (10 mL x 2) to remove excess TFA. The aqueous layer was then freeze-dried. The residue was further purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 1%~20%, 8 min) to obtain HxBz- 22 (0.32 g, 501 umol, yield 73.11%, 2TFA) was obtained as a white solid. 1 H NMR (MeOD, 400 MHz) δ9.20 (s, 2H), 7.84-7.72 (m, 3H), 7.56 (s, 1H), 4.47 (s, 2H), 4.03-3.96 (m, 2H), 3.79 (t, J = 7.2 Hz, 2H), 3.74-3.66 (m, 2H), 3.53-3.36 (m, 2H), 1.88-1.72 (m, 2H), 1.00 (t, J = 7.6 Hz, 3H). LC / MS[M+H]411.2 (calculated value), LC / MS[M+H]411.1 (observed value).

[0230] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[2-hydroxyethoxy(propyl )carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid , Preparation of HxBzL-26a Et 3 N (170mg, 1.68mmol, 234uL, 3.0eq) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-oxo-3-( 2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (396 mg, 560 umol, 1.0 equiv.) was added in one portion at 0 °C. , and then stirred at 0° C. for 0.5 h. The mixture was diluted with water (5ml) and the pH of the mixture was adjusted to about 6 with TFA at 0°C. The aqueous phase was extracted with EtOAc (10 mL) and discarded. The aqueous layer was further extracted with DCM:i-PrOH=3:1 (20 mL x 2). organic layer with Na 2 S.O. 4 Dry on top, filter, and concentrate to give HxBzL-26a (0.53 g, crude, TFA) as a yellow oil.

[0231] Preparation of HxBzL-26 HxBzL-26a (0.35 g, 329 umol, 1.0 eq., TFA) and sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (352 mg, 1.31 EDCI (378 mg, 1.97 mmol, 6.0 eq.) was added in one portion at 25° C. and then stirred at 25° C. for 0.5 h. The mixture was concentrated and filtered. The residue was then purified by preparative HPLC (column: Phenomenex luna C18 250*50mm*10um, mobile phase: [water (0.1% TFA)-ACN], B%: 20%~50%, 10 min). , HxBzL-26 (80.4 mg, 68.2 umol, yield 20.75%) was obtained as a pale yellow oil. 1H NMR (MeOD, 400 MHz) δ9.08 (s, 2H), 7.82-7.70 (m, 3H), 7.56 (s, 1H), 4.69 (s, 2H), 4.06-3.97 (m, 2H), 3.86 (t, J = 6.0 Hz, 2H), 3.83-3.76 (m, 4H), 3.74-3.69 (m, 2H), 3.65-3.57 (m, 36H), 3.46 (s, 2H), 3.02-2.92 (m , 2H), 2.60 (t, J = 6.0 Hz, 2H), 1.87-1.72 (m, 2H), 1.00 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1179.4 (calculated value) , LC / MS[M+H]1179.3 (observed value).

[0232] Example L-30 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[2-(isopropoxycarbonylamino)ethoxy-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy Synthesis of ]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBZL-30 [ka] Isopropyl N-[2-[[2-amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carbonyl]-propyl-amino]oxyethyl ] Preparation of carbamate, HxBz-27a 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBz- in DCM (5 mL) and DMA (3 mL). EDCI (656 mg, 3.42 mmol, 4.0 eq.) in a mixture of 14a (350 mg, 855 umol, 1.0 eq.) and isopropyl N-[2-(propylaminooxy)ethyl]carbamate (268 mg, 1.11 mmol, 1.3 eq., HCl). was added and stirred at 25°C for 1 hour. The mixture was concentrated under reduced pressure at 30°C. The residue was poured into ice water (w / w=1 / 1) (10 mL) and stirred for 5 minutes. Adjust the pH of the mixture to NaHCO 3 It was adjusted to about 8 with an aqueous solution. The aqueous phase was extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (10 mL x 3) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. Purify the residue by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 1 / 1, ethyl acetate / methanol = 1 / 0, 10 / 1). HxBz-27a (460 mg, 772 umol, yield 90.3%) was obtained as a yellow solid. 1 H NMR (MeOD, 400MHz) δ 9.04 (s, 2H), 7.57 (d, J = 8.0 Hz, 1H), 7.51-7.44 (m, 2H), 7.32 (s, 1H), 4.74-4.68 (m, 1H) ), 4.52 (s, 2H), 3.94 (t, J = 5.2 Hz, 2H), 3.73 (t, J = 7.2 Hz, 2H), 3.30-3.26 (m, 2H), 1.76 (sxt, J = 7.2 Hz , 2H), 1.47 (s, 9H), 1.12 (d, J = 6.0 Hz, 6H), 0.98 (t, J = 7.4 Hz, 3H).

[0233] Isopropyl N-[2-[[2-amino-8-[2-(aminomethyl)pyrimidin-5-yl]-3H-1-benzazepine-4-carbonyl]-propyl-amino]oxyethyl]carbamate, HxBz-27 preparation of MeCN (0.5mL) and H 2 To a solution of HxBz-27a (410 mg, 688 umol, 1.0 eq) in O (5 mL) was added TFA (1.18 g, 10.3 mmol, 764 uL, 15.0 eq) and then stirred at 80 °C for 1 h. The mixture was concentrated in vacuo to CH 3 CN was removed. The aqueous phase was extracted with MTBE (5 mL x 3) to remove excess TFA. The aqueous phase was freeze-dried to obtain HxBz-27 (400 mg, 553 umol, 80.3% yield, 2TFA) as a white solid. 1 H NMR (MeOD, 400MHz) δ 9.21 (s, 2H), 7.86-7.74 (m, 3H), 7.51 (s, 1H), 4.76-4.63 (m, 1H), 4.48 (s, 2H), 3.98 (t , J = 5.2 Hz, 2H), 3.77 (t, J = 7.2 Hz, 2H), 3.43 (s, 2H), 1.78 (sxt, J = 7.2 Hz, 2H), 1.12 (d, J = 6.4 Hz, 6H ), 1.00 (t, J = 7.2 Hz, 3H).LC / MS[M+H]496.2 (calculated value), LC / MS[M+H]496.1 (observed value).

[0234] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[2-(isopropoxy carbon ylamino)ethoxy-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Preparation of ethoxy]ethoxy]propanoic acid, HxBzL-30a A solution of HxBz-27 (130mg, 180umol, 1.0eq, 2TFA) in THF (2mL) was added with 3 N (54.5mg, 539umol, 75.0uL, 3.0eq) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-oxo-3- (2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (127 mg, 180 umol, 1.0 eq.) was added at 0 °C; It was then stirred for 0.5 hour. The mixture was concentrated in vacuo. The residue was diluted with water (10 mL) and the pH of the mixture was adjusted to about 6 with TFA. The aqueous phase was extracted with MTBE (5 mL x 3) and discarded. The aqueous phase was further extracted with DCM / i-PrOH=3 / 1 (10 mL x 3). The organic phase was concentrated in vacuo to give HxBzL-30a (180 mg, 174 umol, 96.7% yield) as a yellow oil.

[0235] Preparation of HxBzL-30 HxBzL-30a (180 mg, 174 umol, 1.0 eq.) and sodium (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy (186 mg, 695 umol, EDCI (266 mg, 1.39 mmol, 8.0 eq.) was added to the mixture of 4.0 eq.) and then stirred at 25° C. for 0.5 h. The mixture was concentrated in vacuo and filtered. The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1% TFA)-ACN], B%: 15%~35%, 8 min) to obtain HxBzL. -30 (91 mg, 66.0 umol, yield 38.0%, TFA) was obtained as a yellow solid. 1 H NMR (MeOD, 400MHz) δ 9.08 (s, 2H), 7.82-7.73 (m, 3H), 7.50 (s, 1H), 4.75-4.66 (m, 3H), 3.97 (t, J = 5.2 Hz, 2H ), 3.86 (t, J = 6.0 Hz, 2H), 3.80 (t, J = 6.0 Hz, 2H), 3.75 (br t, J = 7.2 Hz, 2H), 3.66-3.56 (m, 36H), 3.45- 3.42 (m, 2H), 2.96 (t, J = 6.0 Hz, 2H), 2.60 (t, J = 6.4 Hz, 2H), 1.84-1.70 (m, 2H), 1.12 (d, J = 6.0 Hz, 6H) ), 0.99 (t, J = 7.6 Hz, 3H).LC / MS[M+H]1264.4 (calculated value), LC / MS[M+H]1264.7 (observed value).

[0236] Example L-35 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[Propyl-[2-(pyrrolidine-1-carbonylamino)ethoxy]carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-35 [ka] Tert-Butyl N-[[5-[2-amino-4-[propyl-[2-(pyrrolidine-1-carbonylamino)ethoxy]carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl ]Methyl]carbamate, preparation of HxBzL-35a 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBz in DCM (4 mL) and DMA (0.5 mL) -14a (0.25 g, 611 umol, 1.3 eq.) was added to a mixture of N-[2-(propylaminooxy)ethyl]pyrrolidine-1-carboxamide (118 mg, 469 umol, 1.0 eq., HCl) and EDCI (270.12 mg, 1.41 mmol). , 3.0 eq.) were added in one portion at 25°C and then stirred at 25°C for 0.5 h. The mixture was then concentrated and filtered. The mixture was purified by preparative HPLC (column: Phenomenex luna C18 100*40mm*5um, mobile phase: [water (0.1%TFA)-ACN], B%:7%~38%, 8 min) to obtain HxBzL. -35a (0.1 g, 165 umol, yield 35.09%) was obtained as a yellow solid. 1 H NMR (MeOD, 400 MHz) δ9.08 (s, 2H), 7.88-7.68 (m, 3H), 7.50 (s, 1H), 4.54 (s, 2H), 4.02-3.89 (m, 2H), 3.76 (t, J = 7.2 Hz, 2H), 3.44 (s, 2H), 3.36 (t, J = 5.6 Hz, 2H), 3.19-3.07 (m, 4H), 1.86-1.68 (m, 6H), 1.47 ( s, 9H), 1.00 (t, J = 7.6 Hz, 3H).

[0237] 2-amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-propyl-N-[2-(pyrrolidine-1-carbonylamino)ethoxy]-3H-1-benzazepine-4-carboxamide, Preparation of HxBzL-35b H 2 O (4 mL) and CH 3 To a mixture of HxBzL-35a (0.09 g, 148 umol, 1.0 eq.) in CN (0.5 mL) was added TFA (254 mg, 2.23 mmol, 165 uL, 15.0 eq.) in one portion at 25 °C, then at 80 °C for 0.5 h. Stirred. The mixture was then extracted with MTBE (10 mL x 3) and discarded. The aqueous layer was freeze-dried to obtain HxBzL-35b (0.1 g, 136 umol, 91.76% yield, 2TFA) as a yellow solid. 1 H NMR (MeOD, 400 MHz) δ9.21 (s, 2H), 7.86-7.70 (m, 3H), 7.49 (s, 1H), 4.48 (s, 2H), 3.97 (t, J = 5.6 Hz, 2H ), 3.76 (t, J = 7.2 Hz, 2H), 3.48-3.43 (m, 2H), 3.37 (t, J = 5.2 Hz, 2H), 3.13 (s, 4H), 1.81-1.71 (m, 6H) , 1.00 (t, J = 7.6 Hz, 3H).

[0238] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[propyl-[2-( pyrrolidine-1-carbonylamino)ethoxy]carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Preparation of ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-35c Et 3 N (25.0mg, 247umol, 34.4uL, 3.0eq) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-oxo-3- (2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (69.9mg, 98.9umol, 1.2eq) at 0°C. Added in one portion and then stirred at 0° C. for 0.5 h. The mixture was diluted with water (5 mL) and the pH was adjusted to about 6 with TFA at 0°C. The mixture was then extracted with EtOAc (10 mL) and discarded. The aqueous layer was further extracted with DCM:i-PrOH=3:1 (10 mL x 2). organic layer with Na 2 S.O. 4 Dry on top, filter, and concentrate to give HxBzL-35c (0.1 g, crude, TFA) as a yellow oil.

[0239] Preparation of HxBzL-35 Sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate was added to a mixture of HxBzL-35c (0.1 g, 86.1 umol, 1.0 eq., TFA) in DCM (2 mL) and DMA (0.5 mL). (115 mg, 431 umol, 5.0 eq) and EDCI (116 mg, 603 umol, 7.0 eq) were added in one portion at 25°C and then stirred at 25°C for 0.5 h. The mixture was concentrated. The residue was then purified by preparative HPLC (column: Phenomenex Synergi C18 150*25*10um, mobile phase: [water (0.1% TFA)-ACN], B%: 10%~35%, 8 min). , HxBzL-35 (46.4 mg, 33.4 umol, yield 38.78%, TFA) was obtained as a yellow oil. 1 H NMR (MeOD, 400 MHz) δ9.09 (s, 2H), 7.85-7.66 (m, 3H), 7.49 (s, 1H), 4.70 (s, 2H), 3.97 (t, J = 5.6 Hz, 2H ), 3.90-3.84 (m, 2H), 3.80 (t, J = 6.0 Hz, 2H), 3.66-3.58 (m, 38H), 3.45 (s, 2H), 3.37 (t, J = 5.2 Hz, 2H) , 3.13 (s, 4H), 3.01-2.93 (m, 2H), 2.60 (t, J = 6.0 Hz, 2H), 1.86-1.68 (m, 6H), 1.00 (t, J = 7.6 Hz, 3H). LC / MS[M+H]1275.5 (calculated value), LC / MS[M+H]1275.6 (observed value).

[0240] Example L-36 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[1-[[5-[2 -amino-4-[3-(cyclobutoxycarbonylamino)propyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]-3-pyridyl]sulfonyl]azetidin-3-yl]methylamino]-3- Synthesis of oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-36 [ka] Ethyl 2-amino-8-(5-((3-(((tert-butoxycarbonyl)amino)methyl)azetidin-1-yl)sulfonyl)pyridin-3-yl)-3H-benzo[b]azepine-4 - Preparation of carboxylate, HxBz-32b Dioxane (50mL) and H 2 tert-Butyl N-[[1-[[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]sulfonyl] in O (5 mL) azetidin-3-yl]methyl]carbamate, HxBz-32a (5 g, 11.0 mmol, 1 eq) and ethyl 2-amino-8-bromo-3H-1-benzazepine-4-carboxylate (3.41 g, 11.0 mmol, 1 equivalent) in a solution of K 2 C.O. 3 (3.05g, 22.1mmol, 2 equivalents) and Pd(dppf)Cl 2 (403 mg, 551 umol, 0.05 equivalent), N 2 The mixture was added at 25° C. and then stirred at 90° C. for 2 hours. The mixture was filtered and concentrated to give a residue. The residue was diluted with water (100 mL) and extracted with EtOAc (50 mL x 3). The organic layer was washed with brine (50 mL) and 2 S.O. 4 Dry over, filter, and concentrate to obtain HxBz-32a, which was purified by CH at 25 °C for 15 min. 3 Trituration with CN gave HxBz-32b (5.5 g, 9.90 mmol, 89.75% yield) as a gray solid. 1 H NMR (DMSO-d 6 , 400MHz) δ9.29 (s, 1H), 8.94 (s, 1H), 8.32 (s, 1H), 7.80 (s, 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.50-7.41 (m , 2H), 7.04-6.85 (m, 3H), 4.25 (q, J = 7.2 Hz, 2H), 3.82 (t, J = 8.0 Hz, 2H), 3.58-3.52(m, 2H), 2.99-2.85 ( m, 4H), 2.56-2.51 (m, 1H), 1.35-1.30 (m, 12H).

[0241] 2-Amino-8-(5-((3-(((tert-butoxycarbonyl)amino)methyl)azetidin-1-yl)sulfonyl)pyridin-3-yl)-3H-benzo[b]azepine-4- Preparation of carboxylic acid, HxBz-32c MeOH (40mL) and H 2 A solution of HxBz-32b (3.2 g, 5.76 mmol, 1 eq.) in O (5 mL) was added with LiOH.H 2 O (725 mg, 17.3 mmol, 3 eq.) was added and then stirred at 60° C. for 4 hours. The reaction mixture was concentrated under reduced pressure to remove EtOH. The pH of the mixture was adjusted to about 5 with HCl (12M) at 0°C, then filtered and the filter cake was dried under reduced pressure to obtain the crude product. CH the crude product for 20 min at 25 °C. 3Trituration with CN gave HxBz-32c (2.7 g, 5.12 mmol, 88.86% yield) as a gray solid. 1 H NMR (DMSO-d 6 , 400MHz) δ9.34 (s, 1H), 9.02 (s, 1H), 8.42 (s, 1H), 7.98-7.92 (m, 2H), 7.89-7.83 (m, 2H), 3.83 (t, J = 8.0 Hz, 2H), 3.59-3.49 (m, 4H), 2.90 (d, J = 6.0 Hz, 2H), 2.56-2.54 (m, 1H), 1.30 (s, 9H).

[0242] Cyclobutyl N-[3-[[2-amino-8-[5-[3-[(tert-butoxycarbonylamino)methyl]azetidin-1-yl]sulfonyl-3-pyridyl]-3H-1-benzazepine-4 -Carbonyl]-propyl-amino]propyl]carbamate, preparation of HxBz-32d A solution of HxBz-32c (400 mg, 758 umol, 1 eq.) in DMF (10.0 mL) contains HATU (317 mg, 834 umol, 1.1 eq.), DIEA (490 mg, 3.79 mmol, 660 uL, 5 eq.), and cyclobutyl N-[ 3-(Propylamino)propyl]carbamate (380 mg, 1.52 mmol, 2 eq., HCl) was added and stirred at 25° C. for 1 hour. The mixture was diluted with water (50 mL) and extracted with EtOAc (30 mL x 3). The organic layer was washed with brine (20mLx3) and 2 S.O. 4 Dry on top, filter and concentrate. The residue was purified by flash silica gel chromatography (ISCO®, 1 g SepaFlash® silica flash column, eluent of 0-30% ethyl acetate / MeOH @35 mL / min) to give HxBz-32d (340 mg, 469.69 umol, yield 61.95%) was obtained as a pale yellow solid. 1 H NMR (MeOD, 400 MHz) δ9.18 (d, J = 2.0 Hz, 1H), 8.95 (d, J = 2.0 Hz, 1H), 8.42 (t, J = 2.0 Hz, 1H), 7.58-7.50 ( m, 2H), 7.49-7.43 (m, 1H), 6.93 (s, 1H), 4.85-4.76 (m, 1H), 3.90 (t, J = 8.4 Hz, 2H), 3.64-3.56 (m, 2H) , 3.54-3.48 (m, 2H), 3.47-3.39 (m, 2H), 3.32 (br s, 2H), 3.22-3.02 (m, 4H), 2.70-2.57 (m, 1H), 2.35-2.01 (m , 4H), 1.90-1.80 (m, 2H), 1.77-1.47 (m, 4H), 1.37 (s, 9H), 1.05-0.76 (m, 3H).

[0243] Cyclobutyl N-[3-[[2-amino-8-[5-[3-(aminomethyl)azetidin-1-yl]sulfonyl-3-pyridyl]-3H-1-benzazepine-4-carbonyl]-propyl- Preparation of amino]propyl]carbamate, HxBz-32 CH 3 CN (2.00mL) and H 2 To a solution of HxBz-32d (340 mg, 470 umol, 1 eq.) in O (1.00 mL) was added TFA (428 mg, 3.76 mmol, 278 uL, 8 eq.) and then stirred at 80° C. for 1 h. The mixture was concentrated and filtered. The residue was purified by preparative HPLC (column: Phenomenex luna C18 100*40mm*5um, mobile phase: [water (0.1%TFA)-ACN], B%: 5%~35%, 8 min) to obtain HxBz -32 (400 mg, 470 μmol, 99.98% yield, 2TFA) was obtained as a pale yellow solid. 1 H NMR (MeOD, 400 MHz) δ9.24 (d, J = 1.6 Hz, 1H), 9.04 (d, J = 1.6 Hz, 1H), 8.49 (s, 1H), 7.88-7.71 (m, 3H), 7.13 (br s, 1H), 4.85-4.80 (m, 1H), 4.03 (t, J = 8.4 Hz, 2H), 3.73 (dd, J = 5.6, 8.4 Hz, 2H), 3.59-3.43 (m, 4H) ), 3.38 (br s, 2H), 3.12 (br d, J = 7.6 Hz, 4H), 2.83-2.73 (m, 1H), 2.37-2.12 (m, 2H), 2.00-2.10 (m, 4H), 1.78-1.43 (m, 4H), 1.05-0.83 (m, 3H).LC / MS[M+H]624.3 (calculated value), LC / MS[M+H]624.2 (observed value).

[0244] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[1-[[5-[2-Amino-4-[3- (cyclobutoxycarbonylamino)propyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]-3-pyridyl]sulfonyl]azetidin-3-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy] Preparation of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-36a A solution of HxBz-32 (200mg, 235umol, 1eq, 2TFA) in THF (2.00mL) was added with 3 N (71.0mg, 704umol, 98.0uL, 3eq) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-oxo-3- (2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (166 mg, 235 umol, 1 eq) was added and then at 0 °C The mixture was stirred for 1 hour. The mixture was concentrated, diluted with water (10 mL), the pH of the mixture was adjusted to about 6 by gradual addition of TFA, extracted with MTBE (10 mL) and discarded, and the aqueous phase was purified with DCM: It was further extracted with i-PrOH=3:1 (20 mL x 3). organic layer with Na 2 S.O. 4 Dry on top, filter and concentrate to give HxBzL-36a (210 mg, 180.36 umol, 76.81% yield) as a pale yellow oil.

[0245] Preparation of HxBzL-36 HxBzL-33a (210 mg, 180 umol, 1 eq.) and 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonic acid (178 mg, 721 umol, 4 eq.) in DCM (4.00 mL) and DMA (0.20 mL) ) was added EDCI (138 mg, 721 umol, 4 equivalents) and then stirred at 25° C. for 1 hour. The mixture was concentrated and filtered. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.2% FA)-ACN], B%: 15%~40%, 8 min) to obtain HxBzL-36. (98 mg, 68.13 umol, yield 37.77%, FA) was obtained as a white solid. 1 H NMR (MeOD, 400 MHz) δ9.23 (d, J = 2.0 Hz, 1H), 9.02 (d, J = 2.0 Hz, 1H), 8.48 (t, J = 2.0 Hz, 1H), 7.91-7.67 ( m, 3H), 7.13 (s, 1H), 4.85-4.80 (m, 1H), 3.93 (t, J = 8.4 Hz, 2H), 3.86 (t, J = 5.6 Hz, 2H), 3.66-3.55 (m , 40H), 3.54-3.48 (m, 4H), 3.40 (br s, 2H), 3.25-3.08 (m, 4H), 2.97 (t, J = 5.6 Hz, 2H), 2.79-2.68 (m, 1H) , 2.29 (br t, J = 6.0 Hz, 3H), 1.93-1.80 (m, 3H), 1.77-1.52 (m, 4H), 1.01-0.88 (m, 3H).LC / MS[M+H]1392.5 (calculated value), LC / MS[M+H]1392.3 (observed value).

[0246] Example L-37 Cyclobutyl(2-((2-amino-8-(2-(39-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,37-dioxo -6,9,12,15,18,21,24,27,30,33-Decaoxa-2,36-diazanonatriacontyl)pyrimidin-5-yl)-N-propyl-3H-benzo[b] Synthesis of azepine-4-carboxamido)oxy)ethyl)carbamate, HxBzL-37 [ka] Cyclobutyl (2-((2-amino-8-(2-(aminomethyl)pyrimidin-5-yl)-N-propyl-3H-benzo[b]azepine-4-carboxamido)oxy) in DMF (1 ml) ethyl) carbamate, HxBzL-37a (23.6 mg, 0.046 mmol, 1 eq.), and 1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-oxo-7,10 DIPEA (49 μl, 0.28 mmol, 6 (7-azenazotriazotolyloxy-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate), PyAOP, CAS Reg.No.156311-83-0 (59 mg, 0.113 mmol, 2.4 eq.) Added. The reaction was stirred at room temperature for 2 hours, then concentrated and purified by preparative HPLC to yield HxBzL-37 (4.9 mg, 0.0042 mmol, 9%). LC / MS[M+H] 1170.6 (calculated value), LC / MS[M+H] 1170.9 (observed value).

[0247] Example L-41 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]-2-pyridyl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-41 [ka] [ka] Preparation of Tert-Butyl N-[(5-bromo-2-pyridyl)methyl]carbamate, HxBz-36b CH 3 TFA ( 9.19g, 80.6mmol, 5.97mL, 3eq) and Et 3 SiH (31.3g, 268.8mmol, 42.9mL, 10eq) was added at 0°C and stirred at 25°C for 3 hours. The reaction mixture was diluted with Na at 0 °C. 2 C.O. 3 Quenched by adding aqueous solution (200 mL) and concentrated under reduced pressure. The residue was diluted with (200 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (50 mL) and washed with Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure. The residue was purified by column chromatography (SiO 2 , petroleum ether:ethyl acetate=1:0~1:1). HxBz-36b (9 g, crude) was obtained as a pale yellow solid. 1 H NMR (CDCl 3 , 400 MHz) δ8.59 (d, J = 2.4 Hz, 1H), 7.78 (dd, J = 2.4, 8.4 Hz, 1H), 7.20 (d, J = 8.4 Hz, 1H), 5.50 (br s, 1H) ), 4.58-4.29 (m, 2H), 1.45 (s, 9H)

[0248] Preparation of tert-butyl N-[[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridyl]methyl]carbamate, HxBz-36c HxBz-36b (8.00 g, 27.9 mmol, 1 eq.) in dioxane (80 mL), Pin 2 B 2 (8.49g, 33.4mmol, 1.2eq), Pd(dppf)Cl 2 A mixture of (1.02 g, 1.39 mmol, 0.05 eq.) and KOAc (5.47 g, 55.7 mmol, 2 eq.) was degassed with N 2 I purged it 3 times. Mixture, N 2 Stirred under atmosphere at 90° C. for 2 hours and then used directly in the next step without work-up to give HxBz-36c (9.4 g, crude) as a dark brown oil.

[0249] Preparation of ethyl 2-amino-8-[6-[(tert-butoxycarbonylamino)methyl]-3-pyridyl]-3H-1-benzazepine-4-carboxylate, HxBz-36d Dioxane (80mL) and H 2 HxBz-36c (9.30 g, 27.82 mmol, 2 eq.), ethyl 2-amino-8-bromo-3H-1-benzazepine-4-carboxylic acid (4.30 g, 13.9 mmol, 1 eq.) in O (8 mL), Pd(dppf)Cl 2 (509mg, 695umol, 0.05eq), and K 2 C.O. 3 (3.84 g, 27.8 mmol, 2 eq.) was degassed and N 2 Purge 3 times with N 2 Stirred at 90°C for 3 hours under atmosphere. The reaction mixture was filtered and concentrated under reduced pressure. H the residue 2 Diluted with O (50 mL) and extracted with EtOAc (50 mL x 3). Wash the combined organic layers with brine (30 mL x 3) and add Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure. The residue was purified by column chromatography (SiO 2 , petroleum ether:ethyl acetate=1:0~0:1), then (SiO 2 , EtOAc:MeOH=1:0~5:1) to obtain HxBz-36d (2.40 g, 5.50 mmol, yield 39.5%) as a pale yellow solid. 1 H NMR (MeOD, 400 MHz) δ8.76 (s, 1H), 8.10 (br d, J = 8.0 Hz, 1H), 7.85 (s, 1H), 7.58-7.33 (m, 4H), 4.40 (s, 2H), 4.32 (q, J = 7.2 Hz, 2H), 3.05 (s, 2H), 1.48 (s, 9H), 1.38 (t, J = 7.2 Hz, 3H).

[0250] Preparation of 2-amino-8-[6-[(tert-butoxycarbonylamino)methyl]-3-pyridyl]-3H-1-benzazepine-4-carboxylic acid, HxBz-36e A solution of HxBz-36d (2.40 g, 5.50 mmol, 1 eq) in EtOH (30 mL) was added with H 2 LiOH.H in O (6 mL) 2 A solution of O (923 mg, 22.0 mmol, 4 eq.) was added and then stirred at 40° C. for 2 hours. The pH of the reaction mixture was adjusted to 5-6 by adding 1M HCl at 0 °C and then concentrated under reduced pressure to remove EtOH. H the residue 2 Diluted with O (10 mL), filtered, and dried the filter cake under reduced pressure to give HxBz-36e (1.88 g, 4.60 mmol, 83.7% yield) as a gray solid. 1 H NMR (DMSO, 400 MHz) δ9.01 (s, 1H), 8.50 (br d, J = 8.4 Hz, 1H), 7.93 (s, 1H), 7.83 (s, 2H), 7.75 (s, 1H) , 7.73-7.66 (m, 1H), 4.41 (br s, 2H), 3.51 (s, 2H), 1.40 (s, 9H).

[0251] Preparation of Tert-Butyl N-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]-2-pyridyl]methyl]carbamate, HxBz-36f EDCI (493 mg , 2.57 mmol, 3 eq.) and then stirred at 25° C. for 1 hour. The reaction mixture was concentrated under reduced pressure to remove DCM. H the residue 2 Dilute with O (10 mL) and adjust the pH of the mixture to Na at 0 °C. 2 C.O. 3 Adjusted to ~9 by adding aqueous solution and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (5 mL x 3) and washed with Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure. The residue was purified by column chromatography (SiO 2 , petroleum ether:ethyl acetate=1:0~0:1), then (SiO 2 , EtOAc:MeOH=1:0~3:1) to obtain HxBz-36f (0.33 g, 669 umol, yield 78.0%) as a pale yellow solid. 1H NMR (MeOD, 400 MHz) δ8.76 (d, J = 2.0 Hz, 1H), 8.11 (br d, J = 8.4 Hz, 1H), 7.47 (d, J = 8.4 Hz, 2H), 7.43 (d , J = 2.0 Hz, 1H), 7.40-7.34 (m, 1H), 7.29 (s, 1H), 4.40 (s, 2H), 3.95 (q, J = 7.2 Hz, 2H), 3.73 (t, J = 7.2 Hz, 2H), 3.31 (s, 2H), 1.82-1.70 (m, 2H), 1.48 (s, 9H), 1.17 (t, J = 7.2 Hz, 3H), 0.99 (t, J = 7.2 Hz, 3H).

[0252] Preparation of 2-amino-8-[6-(aminomethyl)-3-pyridyl]-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide, HxBz-36 CH 3 CN (3mL) and H 2 To a solution of HxBz-36f (0.33 g, 669 umol, 1 eq) in O (3 mL) was added TFA (610 mg, 5.35 mmol, 396 uL, 8 eq) and then stirred at 80 °C for 0.5 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. H the residue 2 Diluted with O (5 mL), extracted with MTBE (5 mL x 3) and discarded. The aqueous phase was concentrated under reduced pressure to obtain HxBz-36 (0.33 g, 530.95 umol, 79.42% yield, 2TFA) as a pale yellow solid. 1 H NMR (MeOD, 400 MHz) δ8.99 (d, J = 2.0 Hz, 1H), 8.20 (dd, J = 2.4, 8.4 Hz, 1H), 7.79-7.67 (m, 3H), 7.59 (d, J = 8.4 Hz, 1H), 7.45 (s, 1H), 4.36 (s, 2H), 3.98 (q, J = 7.2 Hz, 2H), 3.76 (t, J = 7.2 Hz, 2H), 3.43 (s, 2H) ), 1.83-1.72 (m, 2H), 1.26-1.16 (m, 3H), 1.01 (t, J = 7.2 Hz, 3H).LC / MS[M+H]394.2 (calculated value), LC / MS[ M+H]394.2 (observed value).

[0253] Example L-40 Cyclobutyl(2-((2-amino-8-(2-(38-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,37-dioxo -6,9,12,15,18,21,24,27,30,33-decaoxa-2,36-diazaoctatriacontyl)pyrimidin-5-yl)-N-propyl-3H-benzo[b ]Synthesis of azepine-4-carboxamido)oxy)ethyl)carbamate, HxBzL-40 [ka] Cyclobutyl (2-((2-amino-8-(2-(aminomethyl)pyrimidin-5-yl)-N-propyl-3H-benzo[b]azepine-4-carboxamido)oxy in DMF (0.5 ml) )ethyl) carbamate, HxBzL-40a (12.4 mg, 0.024 mmol, 1 eq.), and 1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-6, DIPEA (25.5 μl , 0.15 mmol, 6 eq.) followed by PyAOP (31.0 mg, 0.059 mmol, 2.4 eq.). The reaction was stirred at room temperature, monitored by LC / MS, then concentrated and purified by preparative HPLC to yield HxBzL-40 (6.7 mg, 0.0058 mmol, 24%). LC / MS[M+H] 1156.6 (calculated value), LC / MS[M+H] 1156.9 (observed value).

[0254] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl] -3H-1-Benzazepin-8-yl]-2-pyridyl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-41a preparation of A solution of HxBz-36 (0.15 g, 241 umol, 1 eq., 2 TFA) in THF (3 mL) contains TEA (73.3 mg, 724 umol, 3 eq.) and 3-[2-[2-[2-[2-[ 2-[2-[2-[2-[2-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Ethoxy]ethoxy]propanoic acid (171 mg, 241 umol, 1 eq) was added at 0°C and stirred at 20°C for 0.5 h. The pH of the reaction mixture was adjusted to 5-6 with TFA at 0 °C, then with H 2 Diluted with O (10 mL), extracted with EtOAc (5 mL x 3) and discarded. The aqueous phase was further extracted with DCM:i-PrOH=3:1 (5mLx3). Mixed organic layer with Na 2 S.O. 4 Dry on top, filter, and concentrate under reduced pressure to give HxBzL-41a (0.23 g, 219 umol, 90.9% yield, TFA) as a colorless oil. 1 H NMR (MeOD, 400 MHz) δ8.91 (d, J = 2.0 Hz, 1H), 8.33 (dd, J = 2.0, 8.0 Hz, 1H), 7.83-7.77 (m, 1H), 7.75-7.69 (m , 3H), 7.47 (s, 1H), 4.64 (s, 2H), 3.98 (q, J = 7.2 Hz, 2H), 3.83-3.74 (m, 4H), 3.71 (t, J = 6.4 Hz, 2H) , 3.66-3.50 (m, 36H), 3.45 (s, 2H), 2.58 (t, J = 6.0 Hz, 2H), 2.53 (t, J = 6.0 Hz, 2H), 1.83-1.73 (m, 2H), 1.21 (t, J = 7.2 Hz, 3H), 1.01 (t, J = 7.2 Hz, 3H)

[0255] Preparation of HxBzL-41 A solution of HxBzL-41a (0.18 g, 172 umol, 1 eq., TFA) in DCM (3 mL) and DMA (0.3 mL) was added with (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy Sodium (184 mg, 687 umol, 4 eq.) and EDCI (132 mg, 687 umol, 4 eq.) were added and stirred at 20° C. for 0.5 h. The reaction mixture was concentrated under reduced pressure to remove DCM and filtered. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 10%~35%, 8 minutes), HxBzL-41 (116.7 mg, 91.4 umol, 53.2% yield, TFA) was obtained as a white solid. 1 H NMR (MeOD, 400 MHz) δ9.01 (d, J = 2.0 Hz, 1H), 8.57 (dd, J = 2.0, 8.4 Hz, 1H), 7.92 (d, J = 8.4 Hz, 1H), 7.84- 7.79 (m, 2H), 7.75-7.68 (m, 1H), 7.45 (s, 1H), 4.72 (s, 2H), 3.98 (q, J = 7.2 Hz, 2H), 3.85 (t, J = 6.0 Hz , 2H), 3.82-3.72 (m, 4H), 3.67-3.51 (m, 36H), 3.45 (s, 2H), 2.96 (t, J = 6.0 Hz, 2H), 2.59 (t, J = 6.0 Hz, 2H), 1.83-1.73 (m, 2H), 1.21 (t, J = 7.2 Hz, 3H), 1.01 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1162.5 (calculated value), LC / MS[M+H]1162.5 (observed value).

[0256] Example L-42 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[3-(cyclobutoxycarbonylamino)propyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]-2-pyridyl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-42 [ka] Cyclobutyl N-[3-[[2-amino-8-[6-[(tert-butoxycarbonylamino)methyl]-3-pyridyl]-3H-1-benzazepine-4-carbonyl]-propyl-amino]propyl] Preparation of carbamate, HxBz-38b 2-Amino-8-[6-[(tert-butoxycarbonylamino)methyl]-3-pyridyl]-3H-1-benzazepine-4-carboxylic acid, HxBz-38a (0.35 g, 857 umol) in DMF (5 mL) , 1 eq.) and cyclobutyl N-[3-(propylamino)propyl]carbamate (258 mg, 1.03 mmol, 1.2 eq., HCl), HATU (326 mg, 857 umol, 1 eq.) and DIEA (332 mg, 2.57 mmol, 448 uL, 3 eq.) was added and then stirred at 20° C. for 2 hours. Add the reaction mixture at 0 °C 2Quenched with O (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (50 mL) and washed with Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, MeOH / ethyl acetate = 1 / 5) to obtain HxBz-38b (0.45 g, 744.12 umol, yield 86.84%) as a yellow solid.

[0257] Cyclobutyl N-[3-[[2-amino-8-[6-[(tert-butoxycarbonylamino)methyl]-3-pyridyl]-3H-1-benzazepine-4-carbonyl]-propyl-amino]propyl] Preparation of carbamate, HxBz-38 MeCN (5 mL) and H 2 To a solution of HxBz-38b (0.45 g, 744 umol, 1 eq) in O (5 mL) was added TFA (679 mg, 5.95 mmol, 441 uL, 8 eq) and stirred at 80 °C for 0.5 h. The reaction mixture was concentrated under reduced pressure to remove MeCN and then extracted with MTBE (5 mL) to remove excess TFA. The aqueous layer was concentrated to obtain a residue, and the residue was subjected to preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 10%~40%, 8 HxBz-38 (0.4 g, 646 umol, 86.89% yield, TFA) was obtained as a yellow solid. 1 H NMR (MeOD, 400 MHz) δ 8.99 (d, J = 1.8 Hz, 1H), 8.20 (dd, J = 2.4, 8.2 Hz, 1H), 7.80-7.66 (m, 3H), 7.59 (d, J = 8.4 Hz, 1H), 7.10 (br s, 1H), 4.85-4.80 (m, 1H), 4.36 (s, 2H), 3.54 (br t, J = 7.2 Hz, 2H), 3.47 (br s, 2H) , 3.36 (br s, 2H), 3.13 (br s, 2H), 2.42-1.96 (m, 2H), 1.92-1.79 (m, 3H), 1.77-1.59 (m, 3H), 0.94 (br s, 3H) ).LC / MS[M+H]505.3 (calculated value), LC / MS[M+H]505.3 (observed value).

[0258] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[3-(cyclobutoxycarbonyl [amino)propyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]-2-pyridyl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy Preparation of ethoxy]propanoic acid, HxBzL-42a A solution of HxBz-38 (0.15g, 204umol, 1eq, 2TFA) in THF (5mL) was added with 3 N (62.1mg, 614umol, 85.49uL, 3eq) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-oxo-3- (2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (145 mg, 205 umol, 1 eq) was added and then at 0 °C The mixture was stirred for 2 hours. Add H to the reaction mixture 2 Quenched with O (5 mL) and adjusted the pH of the mixture to ~6 with TFA, then extracted with EtOAc (10 mL) and discarded, and the aqueous phase was extracted with DCM / PrOH=3 / 1 (20 mL x 3). Further extract with Na 2 S.O. 4 Dry over, filter, and concentrate under reduced pressure to give a residue, HxBzL-42a (0.2 g, 191 umol, 93.46% yield) as a yellow oil.

[0259] Preparation of HxBzL-42 HxBzL-42a (0.2 g, 191 umol, 1 eq) and sodium in DCM (2 mL) and DMA (1 mL); 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (154 mg, 574 umol, EDCI (110 mg, 574 umol, 3 eq.) was added to a solution of 3 eq.) and then stirred at 20° C. for 1 h. The reaction mixture was concentrated under reduced pressure to remove DCM and filtered. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 20%~40%, 8 min) to obtain HxBzL-42. (0.08g, 62.83umol, yield 32.83%) was obtained as a yellow solid. 1 H NMR (MeOD, 400 MHz) δ 9.03 (d, J = 1.8 Hz, 1H), 8.61 (br d, J = 8.4 Hz, 1H), 7.95 (d, J = 8.4 Hz, 1H), 7.87-7.78 ( m, 2H), 7.73 (br s, 1H), 7.11 (s, 1H), 4.73 (s, 3H), 3.85 (t, J = 5.6 Hz, 2H), 3.80 (t, J = 5.6 Hz, 2H) , 3.67-3.50 (m, 38H), 3.64 (br s, 1H), 3.38 (br s, 2H), 3.13 (br s, 2H), 2.95 (t, J = 5.6 Hz, 2H), 2.59 (t, LC / MS[M+H ]1273.5 (calculated value), LC / MS[M+H]1273.7 (observed value).

[0260] Example L-43 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2S)-1-[5 -[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyridine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-43 [ka] Preparation of Tert-butyl (2S)-1-(5-bromopyridine-2-carbonyl)pyrrolidine-2-carboxylate, HxBzL-43a 5-bromopyridine-2-carboxylic acid (2.00g, 9.90mmol, 1.0eq) in DMF (10mL), Et 3 HATU (3.76 g, 9.90mmol, 1.0eq) in N 2 The mixture was stirred at 0°C for 30 min, then heated to 25°C and stirred for a further 0.5 h. Water (30 mL) was added, the aqueous phase was extracted with ethyl acetate (30 mL*3), the combined organic phase was washed with brine (30 mL*1), anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 20 / 1, 2 / 1) to obtain HxBzL-43a (3.40 g, 9.57 mmol, Yield: 96.6%) was obtained as a yellow oil. 1 H NMR (400 MHz, CDCl 3 ) δ8.65 (d, J = 1.6 Hz, 1H), 7.96-7.92 (m, 2H), 5.03 (dd, J = 3.2, 8.4 Hz, 1H), 3.91-3.85 (m, 2H), 2.33-2.28 (m, 2H), 2.18-2.12 (m, 2H), 1.55-1.48 (m, 9H).

[0261] tert-Butyl(2S)-1-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonyl]pyrrolidine-2-carboxylate, HxBzL Preparation of -43b HxBzL-43a (3.40 g, 9.57 mmol, 1.0 eq) in dioxane (30 mL), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2 -dioxaborolan-2-yl)-1,3,2-dioxaborolane (2.92g, 11.5mmol, 1.2eq), Pd(dppf)Cl 2 (700 mg, 957 umol, 0.1 eq.) and AcOK (2.35 g, 23.9 mmol, 2.5 eq.) were degassed and then N 2 The mixture was heated to 100°C for 3 hours. The reaction mixture was concentrated in vacuo to give HxBzL-43b (3.60 g, crude) as a black oil, which was used directly in the next step without purification.

[0262] Preparation of ethyl 2-amino-8-[6-[(2S)-2-tert-butoxycarbonylpyrrolidine-1-carbonyl]-3-pyridyl]-3H-1-benzazepine-4-carboxylate, HxBzL-43c Dioxane (45mL) and H 2 HxBzL-43b (3.60 g, 8.95 mmol, 1.0 eq.), ethyl 2-amino-8-bromo-3H-1-benzazepine-4-carboxylate (2.77 g, 8.95 mmol, 1.0 eq.) in O (5 mL), Pd(dppf)Cl 2 (655 mg, 895 umol, 0.1 equivalent) and K 3P.O. 4 A solution of (3.80g, 17.9mmol, 2.0eq) was degassed and then N 2 The mixture was heated to 95°C for 2 hours under vacuum. Dioxane (45 mL) was removed, the aqueous phase was extracted with ethyl acetate (30 mL*3), the combined organic phases were washed with brine (30 mL*1), and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 10 / 1, 0 / 1) to obtain HxBzL-43c (1.60 g, 3.17 mmol, Yield: 35.4%) was obtained as a pale yellow solid.

[0263] Preparation of 2-amino-8-[6-[(2S)-2-tert-butoxycarbonylpyrrolidine-1-carbonyl]-3-pyridyl]-3H-1-benzazepine-4-carboxylic acid, HxBzL-43d MeOH (10mL) and H 2 A solution of HxBzL-43c (1.60 g, 3.17 mmol, 1.0 equiv) in O (5 mL) was added with LiOH H 2 O (399mg, 9.51mmol, 3.0eq), N 2 was added in one portion at 25° C. and stirred at 25° C. for 10 hours. The reaction mixture was quenched with HCl (4M) until pH=7, then MeOH (10 mL) was removed and the precipitate was filtered and dried to give HxBzL-43d (1.10 g, 2.31 mmol, 72.8% yield) was obtained as a white solid. 1 H NMR (400 MHz, DMSO-d 6 ) δ8.86 (d, J = 2.0 Hz, 1H), 8.32-8.26 (m, 1H), 8.01 (d, J = 8.4 Hz, 1H), 7.95-7.65 (m, 5H), 5.04-5.01 (m , 1H), 3.79-3.82 (m, 2H), 3.52 (s, 2H), 2.36-2.27 (m, 1H), 2.03-1.94 (m, 1H), 1.89-1.77 (m, 2H), 1.45-1.23 (m, 9H).

[0264] Tert-butyl(2S)-1-[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyridine-2-carbonyl]pyrrolidine-2-carboxylate, Preparation of HxBzL-43e EDCI (322 mg, 1.68 mmol, 4.0 equiv), N 2 was added in one portion at 25° C. and then stirred at 25° C. for 1 hour. DCM (4 mL) was removed, water (8 mL) was added, then the pH of the aqueous phase was adjusted to saturated NaHCO 3 8 and extracted with ethyl acetate (5 mL*3), the combined organic phase was washed with brine (5 mL*1), and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100~200 mesh silica gel, petroleum ether / ethyl acetate = 10 / 1, 0 / 1 ~ ethyl acetate / methanol = 10 / 1) to obtain HxBzL. -43e (200 mg, 356 umol, 84.8% yield) was obtained as a brown oil.

[0265] (2S)-1-[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyridine-2-carbonyl]pyrrolidine-2-carboxylic acid, HxBzL-43f preparation of MeCN (1 mL) and H 2 To a solution of HxBzL-43e (200mg, 356umol, 1.0eq) in O (2mL) was added HCl (12M, 890uL, 30eq) with N 2 The mixture was stirred at 80° C. for 1 h and the reaction mixture was concentrated in vacuo to give HxBzL-43f (175 mg, 346 umol, 97.2% yield) as a yellow oil.

[0266] Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2S)-1-[5-[2-amino -4-[Ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyridine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy Preparation of ]ethoxy]ethoxy]propanate, HxBzL-43g HxBzL-43f (175 mg, 346 umol, 1.0 eq.) in DMF (2 mL), tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2- (2-aminoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate (203mg, 346umol, 1.0eq), and Et 3 HATU (132mg, 346umol, 1.0eq) was added to a mixture of N (105mg, 1.04mmol, 145uL, 3.0eq), N 2 Added in one portion at 0° C. and stirred at 0° C. for 30 min, then heated to 25° C. and stirred for an additional 0.5 h. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex luna C18 250*50mm*10um, mobile phase: [water (0.1%TFA)-ACN], B%: 20% to 50%, 10 minutes). Purification provided HxBzL-43g (150 mg, 126 umol, 36.5% yield, TFA) as a pale yellow oil.

[0267] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2S)-1-[5-[2-amino-4- [ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyridine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy Preparation of ethoxy]propanoic acid, HxBzL-43h MeCN (0.2mL) and H 2 To a solution of HxBzL-43g (150mg, 140umol, 1.0eq) in O (2mL) was added HCl (12M, 349uL, 30eq) with N 2 was added in one portion at 25° C. and then stirred at 80° C. for 1 hour. The reaction mixture was concentrated in vacuo to CH 3 CN was removed and the aqueous phase was lyophilized to give HxBzL-43h (140 mg, 137.64 umol, 98.48% yield) as a brown oil.

[0268] Preparation of HxBzL-43 HxBzL-43h (140 mg, 138 umol, 1.0 eq.) and sodium (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy (185 mg, 688 umol) in DCM (1.5 mL) and DMA (0.5 mL). , 5.0 eq.), EDCI (132 mg, 688 umol, 5.0 eq.) was added to a mixture of N 2 was added in one portion at 20° C. and then stirred at 20° C. for 1 hour. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex Luna 80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 10%~40%, 8 minutes). HxBzL-43 (46.3 mg, 35.5 umol, yield 25.8%, purity 95.5%) was obtained as a pale yellow oil. 1 H NMR (400 MHz, MeOD) δ8.96 (d, J = 2.0 Hz, 1H), 8.28 (dd, J = 2.4, 8.4 Hz, 1H), 8.01 (d, J = 8.4 Hz, 1H), 7.84 ( s, 2H), 7.82 (d, J = 1.6 Hz, 1H), 7.48 (s, 1H), 5.16 (dd, J = 4.4, 8.0 Hz, 1H), 4.00 (q, J = 7.2 Hz, 2H), 3.90-3.84 (m, 3H), 3.77 (t, J = 7.2 Hz, 2H), 3.68-3.56 (m, 36H), 3.53-3.43 (m, 6H), 3.22-3.14 (m, 2H), 3.01- 2.96 (m, 2H), 2.42-2.35 (m, 1H), 2.13-1.96 (m, 3H), 1.85-1.75 (m, 2H), 1.23 (t, J = 7.2 Hz, 3H), 1.03 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1245.5 (calculated value), LC / MS[M+H]1245.4 (observed value).

[0269] Example L-45 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2R)-1-[5 -[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-45 [ka] Preparation of (R)-tert-butyl 1-(5-bromopyrimidine-2-carbonyl)pyrrolidine-2-carboxylate, HxBzL-45b A solution of 5-bromopyrimidine-2-carboxylic acid, HxBzL-45a (200 mg, 985 umol, 1 eq) in DMF (3 mL), DIEA (509 mg, 3.94 mmol, 686 uL, 4 eq), and HATU (412 mg, 1.08 mmol, 1.1 eq.) at 0 °C, then stirred for 10 min, and to the mixture was added tert-butyl(2S)-pyrrolidine-2-carboxylate (186 mg, 1.08 mmol, 1.1 eq.), further at 25 °C. Stirred for 3 hours. The reaction mixture was diluted with 20 mL of water and extracted with EtOAc (20 mL x 3). Wash the combined organic layers with brine (20mLx2) and add Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was subjected to column chromatography (SiO 2 , petroleum ether / ethyl acetate=50 / 1 to 1 / 1) to obtain HxBzL-45b (200 mg, 561 umol, yield 56.99%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.18-9.10 (m, 2H), 4.70-4.41 (m, 1H), 3.75-3.48 (m, 2H), 2.42-1.87 (m, 4H), 1.56-1.30 (m, 9H)

[0270] (R)-tert-butyl 1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine-2-carbonyl)pyrrolidine-2-carboxylate, HxBzL -45c preparation HxBzL-45b (200 mg, 561 umol, 1 eq.) and Pin in dioxane (5 mL) 2 B 2 (214 mg, 842 umol, 1.5 eq) was added with KOAc (110 mg, 1.12 mmol, 2 eq) and Pd(dppf)Cl 2 (41.1 mg, 56.2 umol, 0.1 equiv.) with protected N 2 and then stirred at 90° C. for 2 hours. The mixture was filtered and concentrated under reduced pressure. The crude product HxBzL-45c (230 mg, crude) obtained as a brown solid was used in the next step without further purification.

[0271] (R)-tert-butyl 1-(5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyrimidine-2-carbonyl)pyrrolidine-2-carboxy Preparation of HxBzL-45d HxBzL-45c (230 mg, 570 umol, 1 eq.) and 2-amino-8-bromo-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide (209 mg, 570 umol, 1 eq.) in dioxane (5 mL). ) in water (0.2 mL). 2 C.O. 3 (158mg, 1.14mmol, 2eq) and Pd(dppf)Cl 2 A solution of (41.7 mg, 57 umol, 0.1 eq.) was mixed with protected N 2 and then stirred at 90° C. for 16 hours. The mixture was filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate=50 / 1 to ethyl acetate:MeOH=5:1) to obtain HxBzL-45d (240 mg, 427 umol, yield 74.8%) as a yellow oil.

[0272] (R)-1-(5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyrimidine-2-carbonyl)pyrrolidine-2-carboxylic acid, HxBzL -45e preparation H 2 To a solution of HxBzL-45d (240 mg, 427 umol, 1 eq) in O (5 mL) and MeCN (2 mL) was added HCl (12M, 355 uL, 10 eq) and then stirred at 80 °C for 1 h. The mixture was filtered and concentrated under reduced pressure to give HxBzL-45e (170 mg, 336 umol, 78.7% yield) as a yellow oil.

[0273] Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2R)-1-[5-[2-amino -4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy Preparation of ]ethoxy]ethoxy]propanate, HxBzL-45f Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy] in DMF (2 mL) In a solution of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanate (167 mg, 284 umol, 1.2 eq.) and HxBzL-45e (120 mg, 237 umol, 1 eq.) and DIEA (91.9 mg, 711 umol, 124 uL, 3 eq.), HATU (90.1 mg, 237 umol, 1 eq) was added at 0°C, stirred at 0°C for 2 hours, the mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna 80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 20%~45%, 8 min) to obtain HxBzL- 45f (120 mg, 112 umol, yield 47.2%) was obtained as a pale yellow oil.

[0274] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2R)-1-[5-[2-Amino-4- [ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy Preparation of ethoxy]propanoic acid, HxBzL-45g H 2 To a mixture of HxBzL-45f (115 mg, 107 umol, 1 eq) in O (3 mL) was added HCl (12M, 89.2 uL, 10 eq) and then stirred at 80 °C for 1 h. The mixture was filtered and concentrated under reduced pressure to obtain HxBzL-45g (105mg, 103umol, 96.3% yield) as a colorless oil. 1 H NMR (MeOD, 400 MHz) δ9.39-9.04 (m, 2H), 7.88-7.80 (m, 2H), 7.78-7.74 (m, 1H), 7.48 (d, J = 3.0 Hz, 1H), 4.90 -4.62 (m, 1H), 4.03-3.95 (m, 2H), 3.92-3.80 (m, 2H), 3.76 (t, J = 7.2 Hz, 2H), 3.72-3.67 (m, 2H), 3.66-3.57 (m, 38H), 3.48-3.38 (m, 4H), 3.29-3.11 (m, 2H), 2.47 (dt, J = 2.8, 6.2 Hz, 2H), 2.15-1.98 (m, 4H), 1.84-1.73 (m, 2H), 1.44 (s, 9H), 1.22 (t, J = 7.2 Hz, 3H), 1.01 (t, J = 7.4 Hz, 3H).

[0275] Preparation of HxBzL-45 HxBzL-45 g (105 mg, 103 umol, 1 eq.) and sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (111 mg, 413 umol, 4 eq.) in DCM (2 mL) and DMA (0.5 mL) ) was added EDCI (79.1 mg, 413 umol, 4 equivalents) and then stirred at 20° C. for 1 hour. The mixture was filtered and concentrated under residual pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna 80*30mm*3um, mobile phase: [water (0.1% TFA)-ACN], B%: 10%~40%, 8 min) to obtain HxBzL- 45 (60.0 mg, 44.1 umol, 42.8% yield, TFA) was obtained as a pale yellow oil. 1 H NMR (MeOD, 400 MHz) δ9.27-9.21 (m, 2H), 7.89-7.81 (m, 2H), 7.77-7.72 (m, 1H), 7.48-7.44 (m, 1H), 5.05-4.62 ( m, 1H), 3.99 (q, J = 7.0 Hz, 2H), 3.89-3.83 (m, 4H), 3.76 (br t, J = 7.0 Hz, 2H), 3.66-3.53 (m, 36H), 3.50- 3.42 (m, 4H), 3.28-3.20 (m, 2H), 3.16-3.05 (m, 1H), 2.99-2.94 (m, 2H), 2.46-2.26 (m, 1H), 2.12-1.97 (m, 3H) ), 1.82-1.74 (m, 2H), 1.21 (dt, J = 1.8, 7.2 Hz, 3H), 1.04-0.98 (m, 3H).LC / MS[M+H]1246.5 (calculated value), LC / MS[M+H]1246.4 (observed value).

[0276] Example L-46 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2R)-1-[5 -[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyridine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-46 [ka] [ka] Preparation of Tert-butyl (2R)-1-(5-bromopyridine-2-carbonyl)pyrrolidine-2-carboxylate, HxBzL-46b HATU (4.53 g, 11.9 mmol, 1.1 eq.), and Et 3 N (3.29 g, 32.5 mmol, 4.52 mL, 3 eq.) was added followed by tert-butyl (2R)-pyrrolidine-2-carboxylate (2.25 g, 10.8 mmol, 1 eq., HCl). The mixture was stirred at 20°C for 0.5 hour. The reaction mixture was partitioned between EtOAc (150 mL) and water (100 mL). Separate the organic phase and remove Na 2 S.O. 4 Dry on top and concentrate to give a residue. The residue was subjected to column chromatography (SiO 2 , petroleum ether / ethyl acetate = 1 / 0 to 2 / 1) to obtain HxBzL-46b (3.8 g, 10.7 mmol, yield 98.8%) as a yellow oil. 1 H NMR (400MHz, MeOD) δ8.76-8.61 (m, 1H), 8.17-8.13 (m, 1H), 7.91-7.74 (m, 1H), 5.07-4.51 (m, 1H), 3.96-3.67 (m , 2H), 2.43-2.27 (m, 1H), 2.18-1.90 (m, 3H), 1.51 (s, 3H), 1.37 (s, 6H).

[0277] tert-Butyl(2R)-1-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonyl]pyrrolidine-2-carboxylate, HxBzL -46c preparation tert HxBzL-46b (3.5 g, 9.85 mmol, 1 eq.), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3, 2-dioxaborolan-2-yl)-1,3,2-dioxaborolane, Pin 2 B 2 , bis(pinacolato)diboron, CASReg.No.78183-34-3 (3.75g, 14.8mmol, 1.5eq), KOAc (2.42g, 24.6mmol, 2.5eq), Pd(dppf)Cl 2 (721 mg, 985 umol, 0.1 eq.) was added and then stirred at 100° C. for 2 hours. The mixture was used in the next step without work-up and purification. HxBzL-46c (3.96 g, 9.84 mmol, yield 100.00%) was obtained as a black liquid.

[0278] Preparation of ethyl 2-amino-8-[6-[(2R)-2-tert-butoxycarbonylpyrrolidine-1-carbonyl]-3-pyridyl]-3H-1-benzazepine-4-carboxylate, HxBzL-46d Dioxane (100mL) and H 2 HxBzL-46c (3.96 g, 9.84 mmol, 1 eq.), ethyl 2-amino-8-bromo-3H-1-benzazepine-4-carboxylate (3.04 g, 9.84 mmol, 1 eq.) in O (8 mL), Pd(dppf)Cl 2 (360 mg, 492 umol, 0.05 equivalent), and K 2 C.O. 3 A mixture of (3.40g, 24.6mmol, 2.5eq) was stirred at 100°C for 2 hours. The reaction mixture was concentrated to obtain a residue. The residue was dissolved in EtOAc (100 mL) and washed with water (50 mL). Separate the organic phase and remove Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The crude product was subjected to column chromatography (SiO 2 , petroleum ether / ethyl acetate = 1 / 0 to 0 / 1, EA:MeOH = 5:1) to obtain HxBzL-46d (4 g, 7.93 mmol, yield 80.5%) as a yellow solid. 1 H NMR (400MHz, MeOD) δ9.07-8.72 (m, 1H), 8.29-8.16 (m, 1H), 8.12-7.78 (m, 2H), 7.62-7.40 (m, 3H), 5.17-4.47 (m , 1H), 4.34 (q, J = 7.2 Hz, 2H), 4.04-3.75 (m, 2H), 3.67-2.94 (m, 2H), 2.49-2.27 (m, 1H), 2.22-1.88 (m, 3H) ), 1.53 (s, 3H), 1.43-1.34 (m, 9H).

[0279] Preparation of 2-amino-8-[6-[(2R)-2-tert-butoxycarbonylpyrrolidine-1-carbonyl]-3-pyridyl]-3H-1-benzazepine-4-carboxylic acid, HxBzL-46e THF (20mL) and H 2 LiOH.H 2 O (582 mg, 13.9 mmol, 2 eq.) was added and then stirred at 20° C. for 3 hours. The mixture was concentrated to remove THF, then the pH of the mixture was adjusted to about 5 with HCl (4M) and a solid formed from the mixture. The mixture was filtered and the filter cake was dried in vacuo to give HxBzL-46e (3.3 g, 6.93 mmol, 99.8% yield) as a white solid.

[0280] Tert-butyl(2R)-1-[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyridine-2-carbonyl]pyrrolidine-2-carboxylate, Preparation of HxBzL-46f EDCI (483 mg, 2.52 mmol, 3 eq) was added and then stirred at 20°C for 1 hour. The reaction mixture was concentrated to remove DCM and the residue was partitioned between EtOAc (20 mL) and water (20 mL). Separate the organic phase and remove Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was subjected to column chromatography (SiO 2 , petroleum ether / ethyl acetate=1 / 0~0 / 1, EA:MeOH=5:1) to obtain HxBzL-46f (0.32 g, 570 umol, yield 67.9%) as a yellow solid. 1 H NMR (400MHz, MeOD) δ9.06-8.77 (m, 1H), 8.26-8.17 (m, 1H), 8.07-7.87 (m, 1H), 7.53- 7.36 (m, 3H), 7.30 (s, 1H) ), 5.17-4.50 (m, 1H), 4.01-3.69 (m, 6H), 3.01-2.88 (m, 2H), 2.45- 2.30 (m, 1H), 2.18-2.03 (m, 2H), 2.02-1.94 (m, 1H), 1.82-1.73 (m, 2H), 1.52 (s, 3H), 1.36 (s, 6H), 1.18 (t, J = 7.2 Hz, 3H), 1.00 (t, J = 7.2 Hz, 3H).

[0281] (2R)-1-[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyridine-2-carbonyl]pyrrolidine-2-carboxylic acid, HxBzL-46g preparation of H 2 To a mixture of HxBzL-46f (260 mg, 463 umol, 1 eq) in O (5 mL) was added HCl (12M, 579 uL, 15 eq) and then stirred at 80 °C for 1 h. The mixture was concentrated to give HxBzL-46g (0.25g, 461umol, 99.6% yield, HCl) as a yellow oil.

[0282] Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2R)-1-[5-[2-amino -4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin8-yl]pyridine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Preparation of ethoxy]ethoxy]propanate, HxBzL-46h HxBzL-46g (200mg, 369umol, 1eq, HCl) and tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[ To a mixture of 2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate (216mg, 369umol, 1eq), HATU (154mg, 406umol, 1.1eq) and DIEA (143 mg, 1.11 mmol, 193 uL, 3 eq) was added at 0°C and stirred at 0°C for 1 hour. The mixture was concentrated to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 25%~51%, 8 min) to obtain HxBzL-46h. (340 mg, 286 umol, 77.6% yield, TFA) was obtained as a yellow oil.

[0283] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[(2R)-1-[5-[2-Amino-4- [ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyridine-2-carbonyl]pyrrolidine-2-carbonyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy Preparation of ethoxy]propanoic acid, HxBzL-46i H 2 To a mixture of HxBzL-46h (340 mg, 286 umol, 1 eq., TFA) in O (20 mL) was added HCl (12 M, 358 uL, 15 eq.) and then stirred at 80 °C for 0.5 h. The mixture was concentrated to give a residue. The crude product was purified by preparative HPLC (column: Phenomenex Luna 80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 10%~40%, 8 min) to obtain HxBzL. -46i (220 mg, 209 umol, 72.9% yield, HCl) was obtained as a yellow oil.

[0284] Preparation of HxBzL-46 HxBzL-46i (180 mg, 171 umol, 1 eq., HCl) and sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (183 mg, 683 umol, EDCI (164 mg, 854 umol, 5 eq.) was added to a mixture of 4 eq.) and stirred at 15° C. for 0.5 h. The mixture was concentrated to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna 80*30mm*3um, mobile phase: [water (0.1% TFA)-ACN], B%: 15%~45%, 8 min) to obtain HxBzL- 46 (94 mg, 72.6 umol, yield 42.5%, purity 96.2%) was obtained as a colorless oil. 1 H NMR (400MHz, MeOD) δ9.10-8.85 (m, 1H), 8.43-8.16 (m, 1H), 8.11-7.94 (m, 1H), 7.91- 7.71 (m, 3H), 7.48 (s, 1H) ), 5.18-4.65 (m, 1H), 4.07-3.72 (m, 8H), 3.69-3.39 (m, 40H), 3.30- 3.13 (m, 2H), 3.00-2.97 (m, 2H), 2.59-2.23 (m, 1H), 2.19-1.66 (m, 5H), 1.25-1.21 (m, 3H), 1.05-1.00 (m, 3H).LC / MS[M+H]1245.5 (calculated value), LC / MS [M+H]1245.4 (observed value).

[0285] Example L-50(2,3,5,6-tetrafluorophenyl)3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3- [[5-[2-amino-4-[propyl(1H-pyrazol-5-ylmethoxy)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy Synthesis of ]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanate, HxBzL-50 [ka] [ka] Preparation of 5-(chloromethyl)-1H-pyrazole, HxBz-41b A solution of 1H-pyrazol-5-ylmethanol, HxBz-41a (4 g, 40.8 mmol, 1 eq.) in DCM (10 mL) was added with thionyl chloride, SOCl 2 (9.70g, 81.55mmol, 5.92mL, 2 equivalents) was added and then stirred at 0°C to 20°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain HxBz-41b (4.5 g, 38.6 mmol, 94.70% yield) as a white solid. LC / MS[M+H] 117.0 (calculated value), LC / MS[M+H] 117.0 (observed value).

[0286] Preparation of Tert-butyl N-propyl-N-(1H-pyrazol-5-ylmethoxy)carbamate, HxBz-41c To a solution of HxBz-41b (3.01 g, 17.2 mmol, 1 eq.) in DMF (20 mL) at 0 °C was added NaH (1.03 g, 25.7 mmol, 60% purity, 1.5 eq.) and the mixture was heated at this temperature. Stirred for 0.5 h then added KI (285 mg, 1.72 mmol, 0.1 eq.) and 5-(chloromethyl)-1H-pyrazole (2 g, 17.16 mmol, 1 eq.). The resulting mixture was stirred at 20°C for 12 hours. Add NH to the reaction mixture at 0 °C. 4 Quenched with 20 mL of Cl and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (50 mL) and washed with Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex luna C18 (250 * 70 mm, 15 um, mobile phase: [water (0.1% TFA)-ACN], B%: 20% ~ 45%, 20 minutes), HxBz-41c (0.6 g, 2.35 mmol, 13.69% yield) was obtained as a yellow oil. LC / MS [M+H] 256.1 (calculated), LC / MS [M+H] 256.1 (observed).

[0287] Preparation of N-(1H-pyrazol-5-ylmethoxy)propan-1-amine, HxBz-41d MeCN (2 mL) and H 2 To a solution of HxBz-41c (0.5 g, 1.96 mmol, 1 eq) in O (2 mL) was added TFA (2.23 g, 19.58 mmol, 1.45 mL, 10 eq) and then stirred at 80 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove MeCN. The aqueous layer was extracted with 20 mL of MTBE to remove excess TFA. The aqueous layer was lyophilized to give HxBz-41d (0.25 g, crude, TFA) as a yellow oil. 1 H NMR (MeOH, 400 MHz) δ 7.10 (d, J = 2.4 Hz, 1H), 6.47 (d, J = 2.4 Hz, 1H), 5.13 (s, 2H), 3.30-3.20 (m, 2H), 1.78 -1.71 (m, 2H), 1.02 (t, J = 7.2 Hz, 2H).LC / MS[M+H]156.1 (calculated value), LC / MS[M+H]156.1 (observed value).

[0288] Tert-butyl N-[[5-[2-amino-4-[propyl(1H-pyrazol-5-ylmethoxy)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methyl]carbamate, Preparation of HxBz-41f HxBz-41d (0.2 g, 743 umol, 1 eq., TFA salt) and 2-amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidine-5- in DCM (2 mL) and DMA (1 mL). EDCI (854 mg, 4.46 mmol, 6 eq) was added to a solution of HxBz-41e (304 mg, 743 umol, 1 eq), and then stirred at 20 °C for 2 h. did. NaHCO mixture 3 quenched to pH=~8, then extracted with EtOAc (30 mL x 4). The combined organic layers were washed with brine (30 mL) and washed with Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was subjected to column chromatography (SiO 2 , MeOH / ethyl acetate=1 / 5) to obtain HxBz-41f (0.35 g, 640.30 umol, yield 86.19%) as a yellow solid. LC / MS[M+H]547.3 (calculated value), LC / MS[M+H]547.3 (observed value).

[0289] 2-Amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-propyl-N-(1H-pyrazol-5-ylmethoxy)-3H-1-benzazepine-4-carboxamide, HxBz-41 preparation MeCN (2 mL) and H 2 To a solution of HxBz-41f (0.35 g, 640 umol, 1 eq) in O (2 mL) was added TFA (584 mg, 5.12 mmol, 379 uL, 8 eq) and then stirred at 80 °C for 1 h. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna 80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 1%~25%, 8 min) to obtain HxBz- 41 (0.25 g, 371 umol, 57.88% yield, 2TFA) was obtained as a yellow solid. 1 H NMR (MeOH, 400 MHz) δ 9.20 (s, 2H), 7.82-7.78 (m, 1H), 7.74 (d, J = 2.0 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.55 (d, J = 2.0 Hz, 1H), 7.26 (s, 1H), 6.31 (d, J = 2.0 Hz, 1H), 4.96 (s, 2H), 4.48 (s, 2H), 3.80 (t, J = 7.4 Hz, 2H), 3.26 (s, 2H), 1.88-1.73 (m, 2H), 1.01 (t, J = 7.4 Hz, 3H).LC / MS[M+H]447.2 (calculated value), LC / MS[M+H]447.2 (observed value).

[0290] Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-Amino-4-[Propyl (1H -pyrazol-5-ylmethoxy)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Preparation of ethoxy]ethoxy]propanate, HxBzL-50a Et 3 N (90.0mg, 889umol, 124uL, 3eq) and (2,3,5,6-tetrafluorophenyl)3-[2-[2-[2-[2-[2-[2-[2-[ 2-[2-(3-tert-butoxy-3-oxo-propoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate, t-Bu-COO-PEG10-COOTFP (226mg, 296 umol, 1 eq) was added and then stirred at 0°C for 2 hours. Add H to the reaction mixture 2 Quench with 5 mL of O, adjust the pH of the mixture to ~6 with TFA at 0 °C, extract the aqueous phase with EtOAc (10 ml*2) to remove by-products, and extract the aqueous phase with DCM / PrOH=10 / 1 (20mL x 3) and the combined organic phase was extracted with Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give compound HxBzL-50a as a yellow oil. LC / MS[M+H] 1043.56 (calculated value), LC / MS[M+H] 1043.6 (observed value).

[0291] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[propyl(1H-pyrazole- 5-ylmethoxy)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]Propanoic acid, preparation of HxBzL-50b MeCN (2 mL) and H 2 To a solution of HxBzL-50a (0.2 g, 192 umol, 1 eq) in O (2 mL) was added HCl (12M, 320 uL, 20 eq) and then stirred at 80 °C for 2 h. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 5%~35%, 8 min) to obtain HxBzL-50b. (0.13g, 132umol, yield 68.69%) was obtained as a yellow oil. LC / MS[M+H]987.5 (calculated value), LC / MS[M+H]987.6 (observed value).

[0292] Preparation of HxBzL-50 EDCI ( 77.7mg, 405umol, 4eq) was added and then stirred at 20°C for 1 hour. The reaction mixture was filtered. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 20%~40%, 8 min) to obtain HxBzL-50. (0.0216g, 19.0umol, yield 18.78%) was obtained as a yellow solid. 1 H NMR (MeOH, 400 MHz) δ 9.10 (s, 2H), 7.78 (dd, J = 1.6, 8.0 Hz, 1H), 7.71-7.66 (m, 2H), 7.57 (d, J = 2.4 Hz, 1H) , 7.48-7.37 (m, 1H), 7.26 (s, 1H), 7.28-7.24 (m, 1H),6.31 (d, J = 2.4 Hz, 1H), 4.96 (s, 2H), 4.69 (s, 2H) ), 3.86 (t, J = 6.0 Hz, 2H), 3.83-3.76 (m, 4H), 3.68-3.55 (m, 36H), 3.26 (s, 2H), 3.02-2.91 (m, 2H), 2.60 ( t, J = 6.0 Hz, 2H), 1.80 (t, J = 7.2 Hz, 2H), 1.01 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1135.5 (calculated value), LC / MS[M+H]1135.6 (observed value).

[0293] Example L-64 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[2-(1-ethyl-2-oxo-imidazolidin-4-yl)ethyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo Synthesis of -propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-64 [ka] [ka] [ka] Preparation of N-but-3-enyl-4-nitro-N-propyl-benzenesulfonamide, HxBzL-64b Cs 2 C.O. 3 (40.0 g, 123 mmol, 2.5 eq.), KI (8.16 g, 49.1 mmol, 1.0 eq.), and 4-bromobutyl-1-ene (19.9 g, 147 mmol, 15.0 mL, 3.0 eq.) were then added under N2. The mixture was stirred at 40°C for 12 hours. The reaction mixture was poured into ice water (w / w=1 / 1) (150 mL) and stirred for 10 minutes. The aqueous phase was extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with brine (100mLx2) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 10 / 1) to obtain HxBzL-64b (11 g, 36.9 mmol, Yield: 75.1%) was obtained as a yellow solid. 1 H NMR (MeOD, 400MHz)δ 8.51-8.36 (m, 2H), 8.14-7.94 (m, 2H), 5.77-5.70 (m, 1H), 5.10-4.96 (m, 2H), 3.25 (t, J = 7.2 Hz, 2H), 3.15 (t, J = 7.2 Hz, 2H), 2.31 (q, J = 7.2 Hz, 2H), 1.67-1.44 (m, 2H), 0.88 (t, J = 7.2 Hz, 3H) .LC / MS[M+H]299.1 (calculated value), LC / MS[M+H]299.0 (observed value).

[0294] Preparation of 4-nitro-N-[2-(oxiran-2-yl)ethyl]-N-propyl-benzenesulfonamide, HxBzL-64c A solution of HxBzL-64b (13.5 g, 45.3 mmol, 1.0 eq.) in DCM (200 mL) at 0 °C with meta-chloroperbenzoic acid, m-CPBA (18.4 g, 90.5 mmol, 85% purity, 2.0 eq.) was added and then stirred at 20°C for 12 hours. Filter the mixture and dilute the filtrate with saturated NaHSO 3 (30 mL x 1) and brine (100 mL). Organic phase anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 3 / 1) to obtain HxBzL-64c (12 g, 38.2 mmol, yield 84.4%) was obtained as a white solid. LC / MS[M+H]315.1 (calculated value), LC / MS[M+H]315.0 (observed value).

[0295] Preparation of N-[4-(ethylamino)-3-hydroxy-butyl]-4-nitro-N-propyl-benzenesulfonamide, HxBzL-64d To a solution of HxBzL-64c (7 g, 22 mmol, 1.0 eq.) in THF (100 mL) at 0 °C was added ethanamine (33.5 g, 445 mmol, 48.6 mL, 60% purity, 20 eq.), then at 30 °C for 2 Stir for hours. The mixture was concentrated in vacuo at 45°C. The crude product HxBzL-64d (8 g, 22.3 mmol, 99.95% yield) was used in the next step without further purification as a yellow solid. LC / MS[M+H]360.1 (calculated value), LC / MS[M+H]360.2 (observed value).

[0296] Preparation of Tert-Butyl N-ethyl-N-[2-hydroxy-4-[(4-nitrophenyl)sulfonyl-propyl-amino]butyl]carbamate, HxBzL-64e HxBzL-64d (7.6g, 21.1mmol, 1.0eq) in THF (70mL) and H 2 A solution of O (10 mL) was added with NaHCO 3 (3.55g, 42.3mmol, 1.64mL, 2.0eq) and Boc 2O (9.23g, 42.3mmol, 9.71mL, 2.0eq) was added. The mixture was stirred at 25°C for 1 hour. The resulting mixture was poured into ice water (w / w=1 / 1) (50 mL) and stirred for 10 minutes. The aqueous phase was extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (50 mL x 1) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 2 / 1) to obtain HxBzL-64e (8.6 g, 18.7 mmol, Yield: 88.5%) was obtained as a yellow oil. 1 H NMR (MeOD, 400MHz)δ 8.46-8.39 (m, 2H), 8.13-8.04 (m, 2H), 3.78-3.70 (m, 1H),3.39-3.2 (m, 3H), 3.29-3.22 (m, 2H), 3.20-3.14 (m, 2H), 3.10-3.00 (m, 1H), 1.79-1.69 (m, 1H), 1.65-1.53 ​​(m, 3H), 1.45 (s, 9H), 1.09 (t, J = 7.2 Hz, 3H), 0.90 (t, J = 7.2 Hz, 3H).

[0297] Tert-Butyl N-[2-(1,3-dioxoisoindolin-2-yl)-4-[(4-nitrophenyl)sulfonyl-propyl-amino]butyl]-N-ethyl-carbamate, HxBzL-64f preparation of Triphenylphosphine, PPh 3 (4.28 g, 16.3 mmol, 1.5 eq.) and diethyl azodicarboxylate, DEAD (2.84 g, 16.3 mmol, 2.97 mL, 1.5 eq.) were added at 0° C. and stirred at 20° C. for 1 hour. The mixture was poured into ice water (w / w=1 / 1) (50 mL) and stirred for 10 minutes. The aqueous phase was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with brine (30 mL) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 1 / 1) to obtain HxBzL-64f (8.8 g, crude product). was obtained as a yellow solid. LC / MS[M+H]589.2 (calculated value), LC / MS[M+H]589.2 (observed value).

[0298] Preparation of Tert-Butyl N-[2-amino-4-[(4-nitrophenyl)sulfonyl-propyl-amino]butyl]-N-ethyl-carbamate, HxBzL-64g A solution of HxBzL-64f (4.4 g, 7.47 mmol, 1.0 eq.) in MeOH (50 mL) was added with NH at 20 °C. 2 N.H. 2 .H 2 O (2.25g, 44.9mmol, 2.18mL, 6.0eq) was added and then stirred at 80°C for 12 hours. The mixture was filtered and the filtrate was concentrated in vacuo to give HxBzL-64g (3.4g, 7.41mmol, 99.2% yield) as a yellow oil. LC / MS[M+H]459.2 (calculated value), LC / MS[M+H]459.2 (observed value).

[0299] Preparation of N-[3-amino-4-(ethylamino)butyl]-4-nitro-N-propyl-benzenesulfonamide, HxBzL-64h To a solution of HxBzL-64g (2.9g, 6.32mmol, 1.0eq) in EtOAc (30mL) was added HCl / EtOAc (4M, 29.0mL, 18.3eq) and then stirred at 20 °C for 1 h. The mixture was concentrated in vacuo to give HxBzL-64h (2.7 g, crude, 2HCl) as a yellow solid. 1 H NMR (MeOD, 400MHz)δ 8.35 (d, J = 8.8 Hz, 2H), 8.09 (d, J = 8.8 Hz, 2H), 3.78-3.69 (m, 1H), 3.45-3.31 (m, 4H), 3.17-3.05 (m, 4H), 2.12-1.99 (m, 2H), 1.57-1.43 (m, 2H), 1.32 (t, J = 7.2 Hz, 3H), 0.80 (t, J = 7.2 Hz, 3H) .LC / MS[M+H]359.17 (calculated value), LC / MS[M+H]359.1 (observed value).

[0300] Preparation of N-[2-(1-ethyl-2-oxo-imidazolidin-4-yl)ethyl]-4-nitro-N-propyl-benzenesulfonamide, HxBzL-64i HxBzL-64h (2.7g, 7.53mmol, 1.0eq) and Et in THF (30mL) 3 To a mixture of N (1.91 g, 18.8 mmol, 2.62 mL, 2.5 eq) was added carbonyldiimidazole, CDI (2.44 g, 15.1 mmol, 2.0 eq) at 0°C. The mixture was stirred at 25°C for 12 hours. The resulting mixture was poured into ice water (w / w=1 / 1) (50 mL) and stirred for 10 minutes. The aqueous phase was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with brine (50 mL) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 0 / 1) to obtain HxBzL-64i (300 mg, 780 umol, yield 10.4%) was obtained as a yellow oil. 1 H NMR (MeOD, 400MHz)δ 8.39 (d, J = 8.8 Hz, 2H), 8.02 (d, J = 8.8 Hz, 2H), 3.99-3.95 (m, 1H), 3.77-3.63 (t, J = 8.8 Hz, 1H), 3.48-3.38 (m, 1H), 3.35-3.23 (m, 2H), 3.22-3.04 (m, 4H), 1.98-1.74 (m, 2H), 1.66-1.45 (m, 2H), 1.15 (t, J = 7.2 Hz, 3H), 0.87 (t, J = 7.2 Hz, 3H)

[0301] Preparation of 1-ethyl-4-[2-(propylamino)ethyl]imidazolidin-2-one, HxBzL-64j A solution of HxBzL-64i (300mg, 780umol, 1.0eq) in MeCN (10mL) was added with LiOH. 2 O (196 mg, 4.68 mmol, 6.0 eq.) and 2-sulfanylacetate (0.45 g, 4.24 mmol, 384 uL, 5.43 eq.) were added and then stirred at 25° C. for 2 hours. The mixture was filtered and the filtrate was concentrated in vacuo. H the residue 2 diluted with O (20 mL), then the pH of the aqueous phase was adjusted to 3-4 with HCl (1M), then extracted with EtOAc (20 mL x 3) to remove by-products, and then the aqueous phase was freeze-dried to obtain HxBzL-64j (180 mg, 763 umol, 97.8% yield, HCl) as a colorless oil. 1 H NMR (MeOD, 400MHz)δ 3.83-3.73 (m, 1H), 3.65 (t, J = 8.8 Hz, 1H), 3.28-3.13 (m, 3H), 3.12-3.03 (m, 2H), 3.02-2.93 (m, 2H), 2.01-1.84 (m, 2H), 1.79-1.67 (m, 2H), 1.11 (t, J = 7.2 Hz, 3H), 1.03 (t, J = 7.2 Hz, 3H).

[0302] Tert-Butyl N-[[5-[2-amino-4-[2-(1-ethyl-2-oxo-imidazolidin-4-yl)ethyl-propyl-carbamoyl]-3H-1-benzazepine-8- Preparation of yl]pyrimidin-2-yl]methyl]carbamate, HxBzL-64l 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBzL-64k (210 mg, 513 umol) in DMF (6 mL) , 1.0 eq.) were added HATU (205 mg, 539 umol, 1.05 eq.), DIEA (331 mg, 2.56 mmol, 447 uL, 5.0 eq.), and HxBzL-61j (145 mg, 615 umol, 1.2 eq., HCl), then Stirred at 25°C for 1 hour. The resulting mixture was poured into ice water (w / w=1 / 1) (10 mL) and stirred for 5 minutes. The aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (10 mL x 2) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. Purify the residue by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 1 / 0, ethyl acetate / methanol = 1 / 0, 3 / 1). HxBzL-64l (300 mg, 508 umol, yield 99.0%) was obtained as a yellow solid. LC / MS[M+H]591.3 (calculated value), LC / MS[M+H]591.3 (observed value).

[0303] 2-Amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-[2-(1-ethyl-2-oxo-imidazolidin-4-yl)ethyl]-N-propyl-3H- Preparation of 1-benzazepine-4-carboxamide, HxBzL-64m To a solution of HxBzL-64l (300 mg, 508 umol, 1.0 eq.) in EtOAc (5 mL) was added HCl / EtOAc (4M, 6.00 mL, 47.3 eq.) and then stirred at 25° C. for 1 h. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 1%~30%, 8 min) to obtain HxBzL-64m. (142 mg, 198 umol, yield 38.91%, 2TFA) was obtained as a yellow solid. 1 H NMR (MeOD, 400MHz)δ 9.22 (s, 2H), 7.88-7.71 (m, 3H), 7.15 (s, 1H), 4.49 (s, 2H), 3.75-3.60 (m, 2H), 3.57-3.50 (m, 4H), 3.39 (s, 2H), 3.28-3.18 (m, 3H), 2.02-1.97 (s, 1H), 1.88-1.83 (m, 1H), 1.81-1.65 (m, 2H), 1.15 -1.10 (m, 3H), 1.01-0.95 (m, 3H).LC / MS[M+H]491.28 (calculated value), LC / MS[M+H]491.3 (observed value).

[0304] Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[[5-[2-amino-4-[2-( 1-Ethyl-2-oxo-imidazolidin-4-yl)ethyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy] Preparation of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanate, HxBzL-64n HxBzL-64m (90mg, 125umol, 1.0eq, 2TFA) and Et in DMF (1mL) 3 In a solution of N (38.02 mg, 376 umol, 52.3 uL, 3.0 eq.), (2,3,5,6-tetrafluorophenyl)3-[2-[2-[2-[2-[2-[2- [2-[2-[2-(3-tert-butoxy-3-oxo-propoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanate, t-Bu-COO-PEG10- COOTFP (95.5 mg, 125 umol, 1.0 eq) was added at 0°C and then stirred at 25°C for 1 hour. Water (10 mL) was added and then the pH of the mixture was adjusted to about 6 with TFA. The aqueous layer was extracted with MTBE (5 mL x 3) to remove byproducts. The aqueous phase was further extracted with DCM / i-PrOH=3 / 1 (10 mL x 3). The organic phase (DCM / i-PrOH) was concentrated in vacuo to give HxBzL-64n (130 mg, 120 umol, 95.5% yield) as a yellow oil.

[0305] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[2-(1-ethyl -2-oxo-imidazolidin-4-yl)ethyl-propyl-carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy ]Ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, preparation of HxBzL-64o MeCN (0.5mL) and H 2 To a solution of HxBzL-64n (100 mg, 92.0 umol, 1.0 eq) in O (1 mL) was added TFA (83.9 mg, 735 umol, 54.5 uL, 8.0 eq) and then stirred at 80 °C for 1 h. The mixture was concentrated in vacuo to give a residue, the residue was diluted with water (10 mL), the aqueous phase was extracted with MTBE (10 mL) to remove excess TFA, the aqueous phase was lyophilized, HxBzL-64o (100 mg, 87.3 umol, 94.9% yield, TFA) was obtained as a yellow oil.

[0306] Preparation of HxBzL-64 HxBzL-64o (100 mg, 87.3 umol, 1.0 eq., TFA) and sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (70.2 mg, EDCI (67.0 mg, 349 umol, 4.0 eq.) was added to a solution of 262 umol, 3.0 eq.) and then stirred at 20.degree. C. for 1 hour. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 10%~35%, 8 min) to obtain HxBzL-64. (30 mg, 23.8 umol, 27.3% yield) was obtained as a yellow oil. 1 H NMR (MeOD, 400MHz)δ 9.10 (s, 2H), 7.81-7.68 (m, 3H), 7.14 (s, 1H), 4.71 (s, 2H), 3.88 (t, J = 6.0 Hz, 2H), 3.80 (t, J = 6.0 Hz, 2H), 3.68-3.59 (m, 37H), 3.55-3.50 (m, 3H), 3.40 (s, 2H), 3.26-3.20 (m, 3H), 2.98 (t, J = 6.0 Hz, 2H), 2.62 (t, J = 6.0 Hz, 2H), 2.06-1.82 (m, 2H), 1.80-1.67 (m, 2H), 1.15-1.10 (m, 3H), 1.01-0.93 (m, 3H).LC / MS[M+H]1259.5 (calculated value), LC / MS[M+H]1259.6 (observed value).

[0307] Example L-68 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[(1S)-1-[[ (1S)-1-[[4-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylcarbamoyloxymethyl] phenyl]carbamoyl]-4-ureido-butyl]carbamoyl]-2-methyl-propyl]amino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy , 2,3,5,6-tetrafluoro-benzenesulfonic acid, synthesis of HxBzL-68 [ka] [ka] 4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamide)-5-ureidopentanamide)benzyl( Preparation of (5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyrimidin-2-yl)methyl)carbamate, HxBzL-68a 2-Amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide, HxBz-5 (41.2 mg, 96umol, 1 eq., HCl), and Et 3 A solution of N (29.0 mg, 287 umol, 39.9 uL, 3 eq.) was added with (9H-fluoren-9-yl)methyl ((S)-3-methyl-1-(((S)-1- ((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxo-5-uredopentan-2-yl)amino)-1-oxobutan-2-yl)carbamate, Fmoc-Val-Cit -PNC (110 mg, 143 umol, 1.5 eq) was added at 0°C and then stirred at 25°C for 1 hour. Piperidine (24.4 mg, 287 umol, 28.3 uL, 3 eq.) was added to the mixture and stirred for an additional 1 hour at 25°C. The mixture was filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 5% to 30%, 8 minutes), HxBzL-68a (60 mg, 75.0 umol, 78.4% yield) was obtained as a yellow oil. LC / MS[M+H]800.4 (calculated value), LC / MS[M+H]800.6 (observed value).

[0308] Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[(1S)-1-[[(1S)-1 -[[4-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylcarbamoyloxymethyl]phenyl]carbamoyl]- Preparation of 4-ureido-butyl]carbamoyl]-2-methyl-propyl]amino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate, HxBzL-68b A solution of HxBzL-68a (60 mg, 65.7 umol, 1 eq., TFA) in THF (2 mL) was added with Et 3 N (19.9mg, 197umol, 27.4uL, 3eq) and (2,3,5,6-tetrafluorophenyl)3-[2-[2-[2-[2-[2-[2-[2- [2-[2-(3-tert-butoxy-3-oxo-propoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate, t-Bu-COO-PEG10-COOTFP(50.1 mg, 66 umol, 1 eq.) and then stirred at 25° C. for 1 hour. The reaction mixture was diluted with 2 mL of water, then the pH of the aqueous phase was adjusted to 5-6 with TFA, extracted with DCM / i-prOH (5 mL x 3, 3 / 1), and the combined organic phases were diluted with Na 2 S.O. 4Dry over, filter, and concentrate under reduced pressure to give HxBzL-68b (90 mg, 64.4 umol, 98.2% yield) as a yellow oil, which was used in the next step without further purification. LC / MS[M+H] 1396.8 (calculated value), LC / MS[M+H] 1396.7 (observed value).

[0309] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[(1S)-1-[[(1S)-1-[[ 4-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylcarbamoyloxymethyl]phenyl]carbamoyl]-4-ureido -Butyl]carbamoyl]-2-methyl-propyl]amino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, preparation of HxBzL-68c To a solution of HxBzL-68b (90 mg, 64 umol, 1 eq.) in water (3 mL) and MeCN (1 mL) was added TFA (73.5 mg, 644 umol, 47.7 uL, 10 eq.) and then stirred at 80 °C for 2 h. did. The reaction mixture was diluted with 2 mL of water, the pH of the aqueous phase was adjusted to 5-6 with TFA, extracted with DCM / i-prOH (5 mL x 3, 3 / 1), and the combined organic phase was diluted with Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give HxBzL-68c (100 mg, crude) as a yellow oil. LC / MS[M+H] 1340.7 (calculated value), LC / MS[M+H] 1340.6 (observed value).

[0310] Preparation of HxBzL-68 HxBzL-68c (100 mg, 74.6 umol, 1 eq) and sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (80.0 mg, 298 umol, in DCM (1 mL) and DMA (0.5 mL)) EDCI (57.2 mg, 298 umol, 4 eq.) was added to a solution of 4 eq.) and then stirred at 25° C. for 1 h. The mixture was filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 15% to 40%, 8 minutes), HxBzL-68 (20 mg, 12.75 umol, yield 17.09%) was obtained as a white solid. 1 H NMR (400 MHz, MeOD) δ 9.08 (s, 2H), 7.83-7.78 (m, 1H), 7.77-7.71 (m, 2H), 7.65 (br d, J = 7.6 Hz, 2H), 7.47 (s , 1H), 7.42-7.34 (m, 2H), 5.12 (s, 2H), 4.62 (s, 2H), 4.54-4.48 (m, 1H), 4.23-4.18 (m, 1H), 4.02-3.98 (m , 2H), 3.87 (t, J = 6.0 Hz, 2H), 3.80-3.75 (m, 2H), 3.65-3.60 (m, 36H), 3.52-3.49 (m, 2H), 3.47 (s, 2H), 3.21-3.14 (m, 2H), 2.98 (t, J = 6.0 Hz, 2H), 2.61-2.53 (m, 2H), 2.20-2.10 (m, 1H), 2.02-1.88 (m, 1H), 1.85- 1.71 (m, 3H), 1.70-1.52 (m, 2H), 1.23 (t, J = 7.2 Hz, 3H), 1.05-0.99 (m, 9H).LC / MS[M+H]1568.6 (calculated value) , LC / MS [M+H] 1568.6 (observed value).

[0311] Example L-77 4-((1-(5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyrimidin-2-yl)-3- Synthesis of oxo-6,9,12-trioxa-2-azapentadecan-15-oyl)oxy)-2,3,5,6-tetrafluorobenzenesulfonic acid, HxBzL-77 [ka] 1-(5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyridin-2-yl)-3-oxo-6,9,12-trioxa Preparation of -2-azapentadecane-15-acid, HxBzL-77b 2-Amino-8-(2-(aminomethyl)pyrimidin-5-yl)-N-ethoxy-N-propyl-3H-benzo[b]azepine-4-carboxamide, HxBzL-77a in DMF (0.5ml) (25.2 mg, 0.064 mmol, 1 eq.), and 3-(2-(2-(3-((2,5-dioxopyrrolidin-1-yl)oxy)-3-oxopropoxy)ethoxy)ethoxy)propane To a stirred solution of acid (22.2 mg, 0.064 mmol, 1 eq.) was added diisopropylethylamine, DIPEA (56 μl, 0.32 mmol, 5 eq.). The reaction was stirred at room temperature, monitored by LC / MS, and then purified by preparative HPLC to yield HxBzL-77b (28.8 mg, 0.046 mmol, 72%). LC / MS[M+H]627.3 (calculated value), LC / MS[M+H]627.6 (observed value).

[0312] Preparation of HxBzL-77 HxBzL-77b (28.8 mg, 0.046 mmol, 1 eq.) and sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (61.6 mg, 0.23 mmol, 5 eq.) in DMF (0.5 ml) To a stirred solution of was added 2,4,6-collidine (61 μl, 0.46 mmol, 10 eq.) followed by EDC (13.2 mg, 0.069, 1.5 eq.). The reaction was stirred at room temperature, monitored by LC / MS, then concentrated and purified by preparative HPLC to yield HxBzL-77 (8.1 mg, 0.0095 mmol, 21%). LC / MS[M+H]855.3 (calculated value), LC / MS[M+H]855.6 (observed value).

[0313] Example L-78 4-((1-(5-(4-(((1H-pyrazol-4-yl)methyl)(propyl)carbamoyl)-2-amino-3H-benzo[b]azepine-8- yl)pyrimidin-2-yl)-3-oxo-6,9,12,15,18,21,24,27,30,33-decaoxa-2-azahexatriacontan-36-oyl)oxy)-3 Synthesis of ,5-dichlorobenzenesulfonic acid, HxBzL-78 [ka] [ka] Preparation of 4-(chloromethyl)-1H-pyrazole, HxBzL-78a A solution of 1H-pyrazol-4-ylmethanol (2.50 g, 25.5 mmol, 1 eq.) in DCM (40.0 mL) was added with SOCl at 0 °C. 2 (6.06g, 50.9mmol, 3.70mL, 2eq) was added and then stirred at 20°C for 2 hours. The mixture was concentrated to give HxBzL-78a (4 g, crude) as a white solid.

[0314] Preparation of 4-nitro-N-propyl-N-(1H-pyrazol-4-ylmethyl)benzenesulfonamide, HxBzL-78b A solution of Cs 2 C.O. 3 (21.0 g, 64.35 mmol, 3 eq.) and HxBzL-78a (2.5 g, 21.45 mmol, 1 eq.) were added and then stirred at 20° C. for 2 hours. The mixture was diluted with water (50 mL) and extracted with EtOAc (30 mL x 3). The organic layer was washed with brine (20mLx3) and 2 S.O. 4 Dry on top, filter and concentrate. The residue was purified by flash silica gel chromatography (ISCO®, 8 g SepaFlash® silica flash column, eluent with a 0-70% ethyl acetate / petroleum ether gradient at 45 mL / min) and purified by HxBzL-78b ( 2.7 g, 8.32 mmol, yield 38.81%) was obtained as a white solid. 1 H NMR (CDCl 3 , 400 MHz) δ9.05-8.79 (m, 1H), 8.34 (d, J = 8.8 Hz, 2H), 7.97 (d, J = 8.8 Hz, 2H), 7.46 (s, 2H), 4.37 (s, 2H), 3.22-3.10 (m, 2H), 1.52 (sxt, J = 7.2 Hz, 2H), 0.82 (t, J = 7.2 Hz, 3H)

[0315] Preparation of N-(1H-pyrazol-4-ylmethyl)propan-1-amine, HxBzL-78c CH 3 A solution of HxBzL-78b (2.40 g, 7.40 mmol, 1 eq.) in CN (20.0 mL) was added with LiOH.H 2O (1.86 g, 44.40 mmol, 6 eq) and methyl 2-sulfanyl acetate (2.66 g, 25.06 mmol, 2.27 mL, 3.39 eq) were added at 0°C and then stirred at 20°C for 2 hours. The mixture was filtered and concentrated, the residue was diluted with water (40 mL), the pH of the aqueous phase was adjusted to ~1 by gradual addition of HCl (12 M) at 0 °C, and extracted with EtOAc (40 mL). The by-products were removed. The aqueous phase was lyophilized to give HxBzL-78c (1.2 g, 6.83 mmol, 92.33% yield, HCl) as a white solid. 1 H NMR (MeOD, 400 MHz) δ8.28 (s, 2H), 4.26 (s, 2H), 3.04 (br d, J = 7.6 Hz, 2H), 1.77 (qd, J = 7.6, 15.2 Hz, 2H) , 1.03 (t, J = 7.6 Hz, 3H).

[0316] tert-Butyl((5-(4-(((1H-pyrazol-4-yl)methyl)(propyl)carbamoyl)-2-amino-3H-benzo[b]azepin-8-yl)pyrimidin-2-yl ) Methyl)carbamate, preparation of HxBzL-78e 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBzL-78d (300 mg, HATU (293mg, 769.35umol, 1.05eq) and DIEA (474mg, 3.66mmol, 638uL, 5eq) and HxBzL-78c (193mg, 1.10mmol, 1.5eq, HCl) were added to a solution of 732.71umol, 1eq). The mixture was then stirred at 10°C for 1 hour. The mixture was diluted with water (30 mL) and extracted with EtOAc (3 x 30 mL). The organic layer was washed with brine (20mLx3) and 2 S.O. 4 Dry on top, filter and concentrate. The residue was purified by flash silica gel chromatography (ISCO®, 1 g SepaFlash® silica flash column, 0-40% ethyl acetate / MeOH @35 mL / min) to give tert-butyl (( 5-(4-(((1H-pyrazol-4-yl)methyl)(propyl)carbamoyl)-2-amino-3H-benzo[b]azepin-8-yl)pyrimidin-2-yl)methyl)carbamate ( 230 mg, 433.46 umol, yield 59.16%) was obtained as a pale yellow solid. LC / MS[M+H]531.28 (calculated value), LC / MS[M+H]531.2 (observed value).

[0317] N-((1H-pyrazol-4-yl)methyl)-2-amino-8-(2-(aminomethyl)pyrimidin-5-yl)-N-propyl-3H-benzo[b]azepine-4-carboxamide , Preparation of HxBzL-78f To a solution of HxBzL-78e (230 mg, 433.46 umol, 1 eq.) in EtOAc (5.00 mL) was added HCl / EtOAc (4M, 5.00 mL, 46.14 eq.) and then stirred at 15° C. for 1 h. The mixture was concentrated to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 1%~25%, 8 minutes) to obtain HxBzL-78f. (210 mg, 385.66 umol, yield 88.97%, TFA) was obtained as a white solid. 1 H NMR (MeOD, 400 MHz) δ9.20 (s, 2H), 7.83-7.73 (m, 2H), 7.73-7.60 (m, 3H), 7.13 (s, 1H), 4.72-4.58 (m, 2H) , 4.47 (s, 2H), 3.52-3.44 (m, 2H), 3.37 (br s, 2H), 1.78-1.64 (m, 2H), 0.98-0.89 (m, 3H).LC / MS[M+H ]431.2 (calculated value), LC / MS[M+H]431.3 (observed value).

[0318] tert-Butyl 1-(5-(4-(((1H-pyrazol-4-yl)methyl)(propyl)carbamoyl)-2-amino-3H-benzo[b]azepin-8-yl)pyrimidine-2- Preparation of HxBzL-78g A solution of HxBzL-78f (110 mg, 236 umol, 1 eq., HCl) in DMF (2 mL) was added with Et 3 N (72.0mg, 707umol, 98.0uL, 3eq) and (2,3,5,6-tetrafluorophenyl)3-[2-[2-[2-[2-[2-[2-[2- [2-[2-(3-tert-butoxy-3-oxo-propoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate, t-Bu-COO-PEG10-COOTFP (198 mg , 259 umol, 1.1 eq) was added and then stirred at 0°C for 1 hour. The mixture was filtered and purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 30%~50%, 8 min) to obtain HxBzL. -78g (110mg, 96.39umol, yield 40.92%, TFA) was obtained as a colorless oil. LC / MS[M+H] 1027.56 (calculated value), LC / MS[M+H] 1027.5 (observed value).

[0319] 1-(5-(4-(((1H-pyrazol-4-yl)methyl)(propyl)carbamoyl)-2-amino-3H-benzo[b]azepin-8-yl)pyrimidin-2-yl)- Preparation of 3-oxo-6,9,12,15,18,21,24,27,30,33-decaoxa-2-azahexatriacontan-36-acid, HxBzL-78h CH 3 CN (2.00mL) and H 2 To a solution of HxBz-78g (110 mg, 96.39 umol, 1 eq., TFA) in O (1.00 mL) was added TFA (88.0 mg, 771.10 mmol, 57.0 uL, 8 eq.) and then stirred at 80 °C for 1 h. did. The mixture was concentrated to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 5%~35%, 8 min) to obtain HxBzL-78h. (100 mg, 92.16 umol, yield 95.61%, TFA) was obtained as a colorless oil. 1 H NMR (MeOD, 400 MHz) δ9.09 (s, 2H), 7.85-7.63 (m, 5H), 7.13 (s, 1H), 4.69 (s, 2H), 4.62 (s, 2H), 3.80 (t , J = 6.0 Hz, 2H), 3.71 (t, J = 6.4 Hz, 2H), 3.67-3.58 (m, 36H), 3.50-3.44 (m, 2H), 3.36 (s, 2H), 2.60 (t, LC / MS[M+H]971.5 (calculated value) , LC / MS[M+H]971.6 (observed value).

[0320] Preparation of HxBzL-78 In a mixture of HxBzL-78h (110 mg, 113.27 umol, 1 eq.) and (3,5-dichloro-4-hydroxy-benzenesulfonic acid (138 mg, 566 umol, 5 eq.)) in DCM (3 mL) and DMA (0.5 mL). , DIEA (29.8 mg, 226 umol, 2 eq.) and EDCI (108 mg, 566 umol, 5 eq.) in N 2 was added at 25° C. and then stirred at 25° C. for 1 hour. The mixture was filtered and concentrated in vacuo. The residue was purified by preparative HPLC column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 10%~30%, 8 minutes, and HxBzL-78 (20mg , 16.72 umol, yield 14.76%) was obtained as a pale yellow oil. 1 H NMR (MeOD, 400 MHz) δ9.07 (s, 2H), 7.80-7.61 (m, 7H), 7.11 (s, 1H), 4.76-4.58 (m, 4H), 3.86 (t, J = 6.0 Hz , 2H), 3.79 (t, J = 6.0 Hz, 2H), 3.67-3.54 (m, 36H), 3.47 (br t, J= 7.2 Hz, 2H), 3.36 (s, 2H), 2.92 (t, J LC / MS[M+H]1195.4( Calculated value), LC / MS[M+H]1195.4 (observed value).

[0321] Example L-82 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[Propyl-[2-(trifluoromethoxy)ethoxy]carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy Synthesis of ]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-82 [ka] [ka] Preparation of Tert-Butyl N-propyl-N-[2-(trifluoromethoxy)ethoxy]carbamate, HxBzL-82b A mixture of silver triflates contained AgOTf (1.76 g, 6.84 mmol, 3 eq.), 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate), Select F, CAS Reg.No.140681-55-6 (1.21g, 3.42mmol, 1.5eq), KF (530mg, 9.12mmol, 214uL, 4eq), and tert-butyl N-(2-hydroxyethoxy)-N-propyl -carbamate, HxBzL-82a (0.5g, 2.28mmol, 1eq.) in nitrogen gas, N 2 It was added continuously below. Then ethyl acetate EtOAc (15 mL), 2-fluoropyridine, 2-FPy (664 mg, 6.84 mmol, 588 uL, 3 eq.), and trifluoromethyltrimethylsilane, TMSCF 3 (973mg, 6.84mmol, 3eq), N 2 continuous addition at 15°C. The resulting mixture was stirred at 25°C for 12 hours. The reaction mixture was filtered and concentrated under reduced pressure. The residue was subjected to column chromatography (SiO 2 , petroleum ether:MTBE=1:0~10:1) to obtain HxBzL-82b (0.26 g, 905 umol, yield 39.7%) as a colorless oil. 1 H NMR (CDCl 3 , 400 MHz) δ4.21-4.01 (m, 4H), 3.48-3.34 (m, 2H), 1.76-1.59 (m, 2H), 1.50 (s, 9H), 0.92 (t, J = 7.2 Hz, 3H )

[0322] Preparation of N-[2-(trifluoromethoxy)ethoxy]propan-1-amine, HxBzL-82c To a solution of HxBzL-82b (260 mg, 905 umol, 1 eq.) in EtOAc (1 mL) was added HCl / EtOAc (4M, 10 mL, 44.2 eq.). The mixture was stirred at 15°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to give HxBzL-82c (0.26 g, crude, HCl) as a yellow oil. 1 H NMR (MeOD, 400 MHz) δ4.42-4.36 (m, 2H), 4.35-4.29 (m, 2H), 3.34-3.28 (m, 2H), 1.83-1.72 (m, 2H), 1.06 (t, J = 7.2Hz, 3H)

[0323] Tert-Butyl N-[[5-[2-amino-4-[propyl-[2-(trifluoromethoxy)ethoxy]carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methyl] Preparation of carbamate, HxBzL-82e 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBzL- in DCM (5 mL) and DMA (5 mL). To a solution of 82d (330 mg, 805.98 umol, 1 eq.), HxBzL-82c (252 mg, 1.13 mmol, 1.4 eq., HCl) and EDCI (464 mg, 2.42 mmol, 3 eq.) were added at 0 °C, and the mixture was incubated at 15 °C. The mixture was stirred for 1 hour. The reaction mixture was concentrated under reduced pressure to remove DCM. H the residue 2 Dilute with O (20 mL) and aqueous NaHCO 3 The pH was adjusted to 8~9 and extracted with EtOAc (20 mL x 3). Wash the combined organic layers with brine (10mLx3) and add Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2 , petroleum ether:ethyl acetate=1:0~0:1) and then (SiO2, EtOAc:MeOH=1:0~5:1) to obtain a crude product. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna C18 150*30mm*5um, mobile phase: [water (0.1%TFA)-ACN], B%: 20% to 50%, 9 minutes). HxBzL-82e (0.15 g, 217 umol, yield 26.9%, TFA) was obtained as a pale yellow oil. 1 H NMR (MeOD-d 4 , 400 MHz) δ9.08 (s, 2H), 7.82-7.76 (m, 1H), 7.74-7.70 (m, 2H), 7.49 (s, 1H), 4.53 (s, 2H), 4.26-4.14 (m , 4H), 3.79 (t, J = 7.2 Hz, 2H), 3.43 (s, 2H), 1.85-1.75 (m, 2H), 1.48 (s, 9H), 1.01 (t, J = 7.2 Hz, 3H) .LC / MS[M+H]579.25 (calculated value), LC / MS[M+H]579.3 (observed value).

[0324] 2-Amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-propyl-N-[2-(trifluoromethoxy)ethoxy]-3H-1-benzazepine-4-carboxamide, HxBzL-82f preparation of To a solution of HxBzL-82e (0.26 g, 449 umol, 1 eq.) in EtOAc (1 mL) was added HCl / EtOAc (4M, 10 mL, 89.0 eq.) and then stirred at 15° C. for 1 h. The reaction mixture was filtered and the solid was dried under reduced pressure to give HxBzL-82f (185 mg, 336 umol, 74.7% yield, 2HCl) as a yellow solid. 1 H NMR (MeOD-d 4 , 400 MHz) δ9.21 (s, 2H), 7.86-7.79 (m, 2H), 7.78-7.71 (m, 1H), 7.50 (s, 1H), 4.48 (s, 2H), 4.26-4.17 (m , 4H), 3.79 (t, J = 7.2 Hz, 2H), 3.44 (s, 2H), 1.85-1.75 (m, 2H), 1.01 (t, J = 7.2 Hz, 3H).LC / MS[M+ H]479.2 (calculated value), LC / MS[M+H]479.2 (observed value).

[0325] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[propyl-[2-( trifluoromethoxy)ethoxy]carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]Ethoxy]propanoic acid, preparation of HxBzL-82g A solution of HxBzL-82f (80 mg, 145 umol, 1 eq., 2HCl) in DMF (2 mL) was added with TEA (29.4 mg, 290 umol, 40.4 uL, 2 eq.) and 3-[2-[2-[2-[2 -[2-[2-[2-[2-[2-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Ethoxy]ethoxy]ethoxy]propanoic acid, HOOC-PEG10-TFP (103 mg, 145 umol, 1 eq.) was added at 0°C and stirred at 15°C for 1 hour. The pH of the reaction mixture was adjusted to 5-6 with TFA at 0°C, then filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 15% to 50%, 8 minutes), HxBzL-82g (85mg, 75.0umol, 51.7% yield, TFA) was obtained as a pale yellow oil. 1 H NMR (MeOD-d 4 , 400 MHz) δ9.10 (s, 2H), 7.84-7.78 (m, 1H), 7.76-7.70 (m, 2H), 7.50 (s, 1H), 4.69 (s, 2H), 4.27-4.16 (m , 4H), 3.85-3.76 (m, 4H), 3.72 (t, J = 6.4 Hz, 2H), 3.65 (s, 4H), 3.64-3.58 (m, 32H), 3.44 (s, 2H), 2.60 ( t, J = 6.0 Hz, 2H), 2.53 (t, J = 6.4 Hz, 2H), 1.86-1.74 (m, 2H), 1.01 (t, J = 7.2 Hz, 3H).LC / MS[M+H ]1019.47 (calculated value), LC / MS[M+H]1019.5 (observed value).

[0326] Preparation of HxBzL-82 A solution of HxBzL-82g (80mg, 70.6umol, 1eq, TFA) in DCM (3mL) and DMA (0.5mL) was added with (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy Sodium (75.7 mg, 282 umol, 4 eq.) and EDCI (54.1 mg, 282 umol, 4 eq.) were added. The mixture was stirred at 15°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to remove DCM and filtered. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 15% to 40%, 8 minutes), HxBzL-82 (41.1 mg, 30.2 umol, 42.8% yield, TFA) was obtained as a pale yellow oil. 1H NMR (MeOD, 400 MHz) δ9.09 (br s, 2H), 7.85-7.62 (m, 3H), 7.48 (br s, 1H), 4.68 (br s, 2H), 4.24-4.20 (m, 4H) ), 3.91-3.74 (m, 5H), 3.71-3.39 (m, 40H), 2.99-2.93 (m, 2H), 2.64-2.56 (m, 2H), 1.82-1.75 (m, 2H), 1.03-0.96 (m, 3H).LC / MS[M+H]1247.4 (calculated value), LC / MS[M+H]1247.4 (observed value).

[0327] Example L-85 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4 -[Propyl-[2-(2,2,2-trifluoroethoxy)ethoxy]carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy Synthesis of ]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-85 [ka] [ka] Preparation of Tert-Butyl N-propyl-N-[2-(2,2,2-trifluoroethoxy)ethoxy]carbamate, HxBzL-85a A solution of tert-butyl N-(2-hydroxyethoxy)-N-propyl-carbamate (750 mg, 3.42 mmol, 1 eq) in THF (20 mL) was added with sodium hydride, NaH (205 mg, 5.13 mmol, 60% pure). , 1.5 eq.) was added at 0°C. After 30 minutes, 2,2,2-trifluoroethyltrifluoromethanesulfonate (1.19 g, 5.13 mmol, 1.5 eq) was added at 0°C and the mixture was stirred at 15°C for 1 hour. The reaction mixture was dissolved in aqueous NH 4 Quenched by adding Cl (30 mL) at 0 °C and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (10 mL) and washed with Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was subjected to column chromatography (SiO 2 , petroleum ether:MTBE=1:0~5:1) to obtain HxBzL-85a (0.4 g, 1.33 mmol, yield 38.8%) as a pale yellow oil. 1 H NMR (CDCl 3 , 400 MHz) δ4.07-4.01 (m, 2H), 3.92 (q, J = 8.8 Hz, 2H), 3.86-3.79 (m, 2H), 3.46-3.37 (m, 2H), 1.70-1.62 (m , 2H), 1.50 (s, 9H), 0.92 (t, J = 7.2 Hz, 3H)

[0328] Preparation of N-[2-(2,2,2-trifluoroethoxy)ethoxy]propan-1-amine, HxBzL-85b To a solution of HxBzL-85a (150 mg, 498 umol, 1 eq.) in DCM (5 mL) was added methanesulfonic acid, MsOH (144 mg, 1.49 mmol, 106 uL, 3 eq.). The mixture was stirred at 15°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to give HxBzL-85b (0.15 g, crude, MsOH salt) as a pale yellow oil. 1 H NMR (MeOD-d 4 , 400 MHz) δ4.32-4.26 (m, 2H), 4.02 (q, J = 8.8 Hz, 2H), 3.94-3.88 (m, 2H), 3.30-3.28 (m, 2H), 1.85-1.68 (m , 2H), 1.04 (t, J = 7.2 Hz, 3H)

[0329] Tert-Butyl N-[[5-[2-amino-4-[propyl-[2-(2,2,2-trifluoroethoxy)ethoxy]carbamoyl]-3H-1-benzazepin-8-yl]pyrimidine- Preparation of 2-yl]methyl]carbamate, HxBzL-85c 2-Amino-8-[2-[(tert-butoxycarbonylamino)methyl]pyrimidin-5-yl]-3H-1-benzazepine-4-carboxylic acid, HxBzL- in DMA (4 mL) and DCM (4 mL) To a solution of 82d (185 mg, 452 umol, 1 eq.) and HxBzL-85b (175 mg, 587 umol, 1.3 eq., MsOH) was added EDCI (260 mg, 1.36 mmol, 3 eq.) at 0°C. The mixture was stirred at 15°C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove DCM. H the residue 2 Dilute with O (15 mL) and adjust the pH to aqueous Na 2 C.O. 3 The solution was adjusted to about 9 at 0°C and extracted with EtOAc (10 mL x 3). Wash the combined organic layers with brine (5 mL x 3) and add Na 2 S.O. 4 Dry on top, filter and concentrate under reduced pressure to give a residue. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 15% to 50%, 8 minutes), HxBzL-85c (150 mg, 212 umol, 47.0% yield, TFA) was obtained as a pale yellow oil. 1 H NMR (MeOD-d 4 , 400 MHz) δ9.07 (s, 2H), 7.81-7.76 (m, 1H), 7.75-7.69 (m, 2H), 7.50 (s, 1H), 4.53 (s, 2H), 4.15-4.06 (m , 2H), 3.88 (q, J = 8.8 Hz, 2H), 3.82-3.74 (m, 4H), 3.45 (s, 2H), 1.84-1.74 (m, 2H), 1.48 (s, 9H), 1.00 ( t, J = 7.2 Hz, 3H).LC / MS[M+H]593.3 (calculated value), LC / MS[M+H]593.3 (observed value).

[0330] 2-Amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-propyl-N-[2-(2,2,2-trifluoroethoxy)ethoxy]-3H-1-benzazepine-4 -Preparation of carboxamide, HxBzL-85d To a solution of HxBzL-85c (130 mg, 184 umol, 1 eq., TFA) in EtOAc (1 mL) was added HCl / EtOAc (4M, 13.0 mL, 283 eq.). The mixture was stirred at 15°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to give HxBzL-85d (110 mg, crude, 2HCl) as a pale yellow solid. 1 H NMR (MeOD-d 4 , 400 MHz) δ9.21 (s, 2H), 7.85-7.78 (m, 2H), 7.77-7.72 (m, 1H), 7.50 (s, 1H), 4.48 (s, 2H), 4.15-4.08 (m , 2H), 3.89 (q, J = 8.8 Hz, 2H), 3.82-3.74 (m, 4H), 3.46 (s, 2H), 1.84-1.74 (m, 2H), 1.01 (t, J = 7.2 Hz, 3H).LC / MS[M+H]493.2 (calculated value), LC / MS[M+H]493.2 (observed value).

[0331] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[5-[2-amino-4-[propyl-[2-( (2,2,2-trifluoroethoxy)ethoxy]carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylamino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy] Preparation of ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-85e A solution of HxBzL-85d (65 mg, 115 umol, 1 eq., 2HCl) in DMF (2 mL) contains TEA (34.9 mg, 345 umol, 48.0 uL, 3 eq.) and 3-[2-[2-[2-[2 -[2-[2-[2-[2-[2-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Ethoxy]ethoxy]ethoxy]propanoic acid, HOOC-PEG10-TFP (106 mg, 149 umol, 1.3 eq) was added at 0°C and then stirred at 15°C for 0.5 h. The pH of the reaction mixture was adjusted to -6 with TFA at 0°C and filtered. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 10% to 40%, 8 minutes), HxBzL-85e (80 mg, 69.7 umol, 60.7% yield, TFA) was obtained as a yellow oil. 1 H NMR (MeOD-d 4, 400 MHz) δ9.09 (s, 2H), 7.83-7.78 (m, 1H), 7.74 (d, J = 8.8 Hz, 2H), 7.51 (s, 1H), 4.69 (s, 2H), 4.16- 4.08 (m, 2H), 3.89 (q, J = 8.8 Hz, 2H), 3.83-3.75 (m, 6H), 3.72 (t, J = 6.0 Hz, 2H), 3.67-3.56 (m, 36H), 3.46 (s, 2H), 2.60 (t, J = 6.0 Hz, 2H), 2.53 (t, J = 6.0 Hz, 2H), 1.84-1.74 (m, 2H), 1.00 (t, J = 7.2 Hz, 3H) .LC / MS[M+H] 1033.5 (calculated value), LC / MS[M+H] 1033.6 (observed value).

[0332] Preparation of HxBzL-85 A solution of HxBzL-85e (75 mg, 65.4 umol, 1 eq., TFA) in DMA (0.5 mL) and DCM (3 mL) was added with (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxy Sodium (70.1 mg, 262 umol, 4 eq.) and EDCI (62.7 mg, 327 umol, 5 eq.) were added and then stirred at 15° C. for 0.5 h. The reaction mixture was concentrated under reduced pressure to remove DCM, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 15% to 40%, 8 minutes), HxBzL-85 (79.8 mg, 58.0 umol, 88.7% yield, TFA) was obtained as a pale yellow oil. 1 H NMR (MeOD-d 4 , 400 MHz) δ9.09 (s, 2H), 7.82-7.68 (m, 3H), 7.49 (s, 1H), 4.69 (s, 2H), 4.14-4.08 (m, 2H), 3.94-3.84 (m , 4H), 3.83-3.74 (m, 6H), 3.69-3.55 (m, 36H), 3.46 (s, 2H), 2.97 (t, J = 6.0 Hz, 2H), 2.60 (t, J = 6.0 Hz, 2H), 1.85-1.73 (m, 2H), 1.00 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1261.4 (calculated value), LC / MS[M+H]1261.6 (observed value ).

[0333] 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[(1S)-2-[(2S)-2- [[(1S)-1-[[4-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylcarbamoyloxy methyl]phenyl]carbamoyl]-2-methyl-propyl]carbamoyl]pyrrolidin-1-yl]-1-methyl-2-oxo-ethyl]amino]-3-oxo-proxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy Synthesis of ]ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-91 [ka] [4-[[(2S)-2-[[(2S)-1-[(2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)propanoyl]pyrrolidine-2-carbonyl]amino]-3 -Methyl-butanoyl]amino]phenyl]methyl N-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methyl]carbamate , Preparation of HxBzL-91a 2-Amino-8-[2-(aminomethyl)pyrimidin-5-yl]-N-ethoxy-N-propyl-3H-1-benzazepine-4-carboxamide in DMF (5 mL), HxBz-5 (0.15 g , 348.08 umol, 1 eq., HCl) and [4-[[(2S)-2-[[(2S)-1-[(2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)propanoyl] Et 3 N (106 mg, 1.04 mmol, 145 uL, 3 eq.) was added. The mixture was stirred at 20°C for 2 hours. Piperidine (74.2 mg, 871 umol, 86.0 uL, 3 eq.) in DMF (1 mL) was then added. The mixture was stirred for an additional hour at 20°C. The reaction mixture was filtered. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 15%~45%, 8 min) to obtain HxBzL-91a. (0.12 g, 148 umol, yield 50.96%) was obtained as a white solid. LC / MS[M+H]811.4 (calculated value), LC / MS[M+H]811.6 (observed value).

[0334] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[[(1S)-2-[(2S)-2-[[( 1S)-1-[[4-[[5-[2-amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrimidin-2-yl]methylcarbamoyloxymethyl]phenyl ]carbamoyl]-2-methyl-propyl]carbamoyl]pyrrolidin-1-yl]-1-methyl-2-oxo-ethyl]amino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Preparation of ethoxy]ethoxy]ethoxy]propanoic acid, HxBzL-91b A solution of HxBzL-91a (0.12 g, 148 umol, 1 eq) in DMF (5 mL) was added with Et 3 N (44.9mg, 444umol, 61.8uL, 3eq) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-oxo-3- (2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, HOOC-PEG10-TFP (105mg, 148umol, 1 eq) was added and then stirred for 1 hour at 20°C. The reaction mixture was filtered and purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1% TFA)-ACN], B%: 20%~50%, 8 min), HxBzL-91b (0.15 g, 111 umol, yield 75.00%) was obtained as a white solid. LC / MS[M+H] 1351.7 (calculated value), LC / MS[M+H] 1351.8 (observed value).

[0335] Preparation of HxBzL-91 HxBzL-91b (0.15 g, 111 umol, 1 eq.) and (2,3,5,6-tetrafluoro-4-hydroxy-phenyl)sulfonyloxysodium (89.3 mg, 333 umol) in DCM (5 mL) and DMA (1 mL) , 3 eq.) was added EDCI (63.8 mg, 333 umol, 3 eq.). The mixture was stirred at 20°C for 2 hours. The reaction mixture was filtered and purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1% TFA)-ACN], B%: 20%~45%, 8 min), HxBzL-91 (0.11 g, 69.6 umol, yield 62.74%) was obtained as a yellow solid. 1 H NMR (MeOH, 400 MHz) δ 9.06 (s, 2H), 7.83-7.69 (m, 3H), 7.59 (d, J = 8.4 Hz, 1H), 7.45 (s, 1H), 7.36 (d, J = 8.4 Hz, 1H), 5.09 (s, 2H), 4.66-4.57 (m, 4H), 4.35-4.18 (m, 1H), 3.98 (q, J = 6.8 Hz, 2H), 3.85 (br t, J = 6.0 Hz, 2H), 3.81-3.68 (m, 5H), 3.67-3.54 (m, 37H), 3.45 (s, 2H), 2.96 (t, J = 5.6 Hz, 2H), 2.56-2.44 (m, 2H) ), 2.22-2.15 (m, 2H), 2.05-2.00 (m, 2H), 1.87-1.71 (m, 2H), 1.44-1.28 (m, 3H), 1.21 (br t, J = 7.2 Hz, 3H) , 1.09-0.85 (m, 9H).LC / MS[M+H]1579.65 (calculated value), LC / MS[M+H]1579.6 (observed value).

[0336] Example L-96 2-amino-8-(2-((1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-6,9,12, 15,18,21,24,27,30-Nonaoxa-3-azadotriacontan-32-yl)carbamoyl)pyrimidin-5-yl)-N-ethoxy-N-propyl-3H-benzo[b]azepine-4 -Synthesis of carboxamide, HxBzL-96 [ka] 2-Amino-8-(2-((29-amino-3,6,9,12,15,18,21,24,27-nonaoxanonacosyl)carbamoyl)pyrimidin-5-yl)-N-ethoxy -N-Propyl-3H-benzo[b]azepine-4-carboxamide, preparation of HxBzL-96b 5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl)pyrimidine-2-carboxylic acid, HxBzL-96a (0.0106g, 0.026 mmol, 1 eq) and tert-butyl (29-amino-3,6,9,12,15,18,21,24,27-nonaoxanonacosyl)carbamate (0.014 g, 0.026 mmol, 1 eq). The solution was added with triethylamine, TEA (36 μl, 0.26 mmol, 10 eq.) followed by (7-azabenzotriazol-1-yloxy) tripyrrolidoline phosphonium hexaphorphosphate, PyAOP, CAS Reg. No. 156311-83-0 ( 0.013g, 0.026mmol, 1 eq) was added. The reaction was stirred at room temperature and monitored by LC / MS. The reaction was concentrated and purified by preparative HPLC to give tert-butyl (1-(5-(2-amino-4-(ethoxy(propyl)carbamoyl)-3H-benzo[b]azepin-8-yl) pyrimidin-2-yl)-1-oxo-5,8,11,14,17,20,23,26,29-nonaoxa-2-azahentriacontan-31-yl)carbamate, then in TFA. and concentrated to obtain HxBzL-96b (17.1 mg, 0.020 mmol, 77%). LC / MS[M+H]848.5 (calculated value), LC / MS[M+H]848.8 (observed value).

[0337] Preparation of HxBzL-96 A solution of HxBzL-96b (17.1 mg, 0.020 mmol, 1.33 eq.) in DMF (0.5 ml) was added with TEA (28 μl, 0.20 mmol, 13.3 eq.) followed by N-(α-maleimidoacetoxy)succinimide ester (3.8 mg , 0.015 mmol, 1 eq) was added. The reaction was stirred at room temperature, monitored by LC / MS, then concentrated and purified by preparative HPLC to yield HxBzL-96 (8.5 mg, 0.0086 mmol, 57%). LC / MS[M+H]985.5 (calculated value), LC / MS[M+H]985.6 (observed value).

[0338] Example L-104 1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-6,9,12,15,18,21,24,27,30 ,33-Decaoxa-3-azapentatriacontan-35-yl(2-((2-amino-N-propyl-8-(pyrimidin-5-yl)-3H-benzo[b]azepine-4-carboxamide) Synthesis of oxy)ethyl)carbamate, HxBzL-104 [ka] Preparation of 2-amino-N-(2-aminoethoxy)-N-propyl-8-(pyrimidin-5-yl)-3H-benzo[b]azepine-4-carboxamide, HxBzL-104b tert-Butyl (2-((2-amino-N-propyl-8-(pyrimidin-5-yl)-3H-benzo[b]azepine-4-carboxamido)oxy)ethyl)carbamate, HxBzL-104a (5.1 mg , 0.011 mmol, 1 eq.) was suspended in 100 μl of TFA. After 30 minutes, the solution was concentrated to give HxBzL-104b (7.4 mg, 0.012 mmol, 100%) as a TFA salt. LC / MS[M+H]381.2 (calculated value), LC / MS[M+H]381.3 (observed value).

[0339] Preparation of HxBzL-104 A solution of HxBzL-104b (7.4 mg, 0.012 mmol, 1 eq.) in DMF (0.5 ml) contains triethylamine (13.5 μl, 0.097 mmol, 8 eq.) and 1-(2,5-dioxo-2,5- Dihydro-1H-pyrrol-1-yl)-2-oxo-6,9,12,15,18,21,24,27,30,33-decaoxa-3-azapentatriacontan-35-yl(4- Nitrophenyl) carbonate, Mal-PEG10-PNC (15.6 mg, 0.019 mmol, 1.6 eq.) was added. The solution was stirred at room temperature and monitored by LC / MS. Upon consumption of the amine starting material, the reaction was concentrated and purified by preparative HPLC to yield HxBzL-104. LC / MS[M+H] 1045.5 (calculated value), LC / MS[M+H] 1045.8 (observed value).

[0340] Example L-107 1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-6,9,12,15,18,21,24,27,30 ,33-Decaoxa-3-azapentatriacontan-35-yl(2-((2-amino-8-(4-methylpyridin-3-yl)-N-propyl-3H-benzo[b]azepine-4 -Synthesis of carboxamido)oxy)ethyl)carbamate, HxBzL-107 [ka] Preparation of 2-amino-N-(2-aminoethoxy)-8-(4-methylpyridin-3-yl)-N-propyl-3H-benzo[b]azepine-4-carboxamide, HxBzL-107b Tert-butyl (2-((2-amino-8-bromo-N-propyl-3H-benzo[b]azepine-4-carboxamido)oxy)ethyl)carbamate in 10:1 dioxane:water, HxBzL-107a (30 mg , 0.062 mmol, 1 eq), (4-methylpyridin-3-yl)boronic acid (17 mg, 0.125 mmol, 2 eq), potassium carbonate (54 mg, 0.39 mmol, 6.3 eq), and [1.1′-bis(diphenyl) A suspension of dichloropalladium(II) (2.3 mg, 0.0032 mmol, 0.05 eq.) was stirred at 80° C. and monitored by LC / MS. The reaction mixture was concentrated and purified by reverse phase column chromatography to give tert-butyl(2-((2-amino-8-(4-methylpyridin-3-yl)-N-propyl-3H-benzo[b ]Azepine-4-carboxamido)oxy)ethyl)carbamate, which was dissolved in minimal TFA and then concentrated to yield HxBzL-107b (12.4 mg, 0.032 mmol, 52%). LC / MS[M+2H] 197.6 (calculated value), LC / MS[M+2H] 197.9 (observed value).

[0341] Preparation of HxBzL-107 HxBzL-107b (12.4 mg, 0.32 mmol, 1 eq.) and 1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-6 in DMF (0.5 ml) ,9,12,15,18,21,24,27,30,33-decaoxa-3-azapentatriacontan-35-yl(4-nitrophenyl) carbonate (25.3 mg, 0.32 mmol, 1 eq.) To the solution was added TEA (22 μl, 0.16 mmol, 5 eq.). The reaction was stirred at room temperature, monitored by LC / MS, then concentrated and purified by preparative HPLC to yield HxBzL-107 (6.1 mg, 0.0058 mmol, 18%). LC / MS[M+H] 1058.5 (calculated value), LC / MS[M+H] 1058.7 (observed value).

[0342] Example L-114 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[1-[2-Amino-4 -[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]pyrazol-4-yl]sulfonylamino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoyl Synthesis of [oxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-114 [ka] Preparation of 4-benzylsulfanyl-1H-pyrazole, HxBzL-114a To a solution of 4-bromo-1H-pyrazole (2.00 g, 13.6 mmol, 1.0 eq) in THF (20 mL) was added n-BuLi (2.50 M, 17.9 mL, 3.3 eq) dropwise at 0 °C. After stirring the mixture at 20°C for 1 hour, (benzyldisulfanyl)methylbenzene (3.35g, 13.6mmol, 1.0 eq.) was added and the mixture was stirred at 0°C for an additional 2 hours. Place reactants on ice / H 2 Quench slowly with O (10 mL) and adjust pH to ~6 with 2N HCl. The mixture was then extracted with EtOAc (20 mL*3). The combined organic phases were washed with brine (50 mL) and anhydrous Na 2 S.O. 4 Dry on top, filter and concentrate in vacuo. The residue was purified by flash silica gel chromatography (ISCO®, 40 g SepaFlash silica flash column, El gradient 50 mL / min from 0 to 30% ethyl acetate / petroleum ether gradient) to give HxBzL-114a (1.70 g, 8.93 mmol). , yield 65.6%) as a yellow solid. 1 H NMR (CDCl 3, 400MHz) δ7.31 (s, 2H), 7.21-7.17 (m, 3H), 7.07-7.06 (m, 2H), 3.73 (s, 2H)

[0343] Preparation of 8-(4-benzylsulfanylpyrazol-1-yl)-N-ethoxy-N-propyl-2-(tritylamino)-3H-1-benzazepine-4-carboxamide, HxBzL-114c 8-bromo-N-ethoxy-N-propyl-2-(tritylamino)-3H-1-benzazepine-4-carboxamide, HxBzL-114b (300 mg, 492 umol, 1.0 eq.) and HxBzL-114a in dioxane (6 mL). (187mg, 985umol, 2.0eq) of Cs 2 C.O. 3 (321mg, 985umol, 2.0eq), [(2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1 ,1'-biphenyl)]palladium(II) methanesulfonate, tBuXPhos Pd G3, CAS Reg.No.1447963-75-8 (117 mg, 147 umol, 0.3 eq.), and di-tert-butyl-[2-( 2,4,6-triisopropylphenyl)phenyl]phosphane, tBuXPhos, CAS Reg.No.564483-19-8 (62.8mg, 147umol, 0.3eq.), N 2 was added in one portion at 20° C. and then stirred at 120° C. for 15 hours. The mixture was poured into water (10 mL). The aqueous phase was extracted with ethyl acetate (5mL*3). The combined organic phases were washed with brine (10 mL) and anhydrous Na 2 S.O. 4 dried, filtered and concentrated in vacuo. The residue was purified by flash silica gel chromatography (ISCO®, 20 g SepaFlash silica flash column, eluent with 0-50% ethyl acetate / petroleum ether gradient 40 mL / min) to give HxBzL-114c (0.300 g, 417 umol, Yield: 84.7%) was obtained as a yellow solid. LC / MS[M+H]718.3 (calculated value), LC / MS[M+H]718.4 (observed value).

[0344] Preparation of 1-[4-[ethoxy(propyl)carbamoyl]-2-(tritylamino)-3H-1-benzazepin-8-yl]pyrazole-4-sulfonyl chloride, HxBzL-114d H 2 To a mixture of HxBzL-114c (300 mg, 417 umol, 1.0 eq.) in O (0.7 mL), AcOH (2 mL) was added N-chlorosuccinimide, NCS (223 mg, 1.67 mmol, 4.0 eq.), then at 20 °C. Stirred for 1 hour. Pour the mixture into ice water (w / w=1 / 1) (10 mL) and dilute with NaHCO 3 The pH was adjusted to approximately 7 with an aqueous solution. The aqueous phase was extracted with ethyl acetate (10 mL*3). The combined organic phases were washed with brine (15 mL) and anhydrous Na 2 S.O. 4 , filtered, and concentrated in vacuo to give HxBzL-114d (0.300 g, crude) as a yellow oil. LC / MS[M+H]694.2 (calculated value), LC / MS[M+H]694.2 (observed value).

[0345] Tert-Butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[1-[4-[ethoxy(propyl)carbamoyl]- 2-(Tritylamino)-3H-1-benzazepin-8-yl]pyrazol-4-yl]sulfonylamino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate, HxBzL- Preparation of 114e HxBzL-114d (0.30 g, 432 umol, 1 eq.) and tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2 Et 3 N (65.5 mg, 648 umol, 90.2 uL, 1.5 eq) was added at 0°C and then stirred at 20°C for 1 hour. The mixture was poured into ice water (w / w=1 / 1) (10 mL). The aqueous phase was extracted with DCM (5mL*3). The combined organic phases were washed with brine (15 mL) and anhydrous Na 2 S.O. 4 , filtered and concentrated in vacuo to give HxBzL-114e (570 mg, crude) as a yellow oil. LC / MS[M+H]1243.6 (calculated value), LC / MS[M+H]1243.6 (observed value).

[0346] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[[1-[2-amino-4-[ethoxy(propyl)carbamoyl] -3H-1-Benzazepin-8-yl]pyrazol-4-yl]sulfonylamino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid, preparation of HxBzL-114f To a mixture of HxBzL-114e (280 mg, 225 umol, 1.0 eq) in DCM (3 mL) was added TFA (770 mg, 6.76 mmol, 500 uL, 30 eq) at 20 °C and then stirred at 50 °C for 15 h. The mixture was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 10%~35%, 8 min) to obtain HxBzL-114f. (50.0 mg, 52.9 umol, yield 23.5%) was obtained as a white solid. 1 H NMR (MeOD , 400MHz) δ8.90 (s, 1H), 8.07 (s, 1H), 7.97 (d, J = 2.0 Hz, 1H), 7.93 (dd, J = 2.0, 8.8 Hz, 1H), 7.73 (d, J = 8.8 Hz, 1H), 7.45 (s, 1H), 3.98 (q, J = 7.2 Hz, 2H), 3.80-3.67 (m, 4H), 3.65-3.51 (m, 38H), 3.43 (s, 2H), 3.19 (t, J = 5.2 Hz, 2H), 2.53 (t, J = 6.4 Hz, 2H), 1.84-1.72 (m, 2H), 1.21 (t, J = 7.2 Hz, 3H), 1.00 (t, J = 7.2 Hz, 3H).LC / MS[M+H]945.4 (calculated value), LC / MS[M+H]945.5 (observed value).

[0347] Preparation of HxBzL-114 Sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (56.7 mg, 211 umol, 4.0 eq.) and EDCI (50.7 mg, 264 umol, 5.0 eq.) were added in one portion at 25°C and then stirred at 25°C for 0.5 h. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 15%~50%, 8 min) to obtain HxBzL-114. (27.8 mg, 23.7 umol, yield 44.7%) was obtained as a white solid. 1 H NMR (MeOD , 400MHz) δ8.89 (s, 1H), 8.05 (s, 1H), 7.96 (d, J = 2.0 Hz, 1H), 7.92 (dd, J = 2.0, 8.4 Hz, 1H), 7.71 (d, J = 8.4 Hz, 1H), 7.44 (s, 1H), 3.97 (q, J = 7.2 Hz, 2H), 3.85 (t, J = 5.6 Hz, 2H), 3.75 (t, J = 7.2 Hz, 2H), 3.68 -3.50 (m, 38H), 3.45 (s, 2H), 3.19 (t, J = 5.6 Hz, 2H), 2.96 (t, J = 6.0 Hz, 2H), 1.77 (t, J = 7.2 Hz, 2H) , 1.20 (t, J = 7.2 Hz, 3H), 1.00 (t, J = 7.2 Hz, 3H).LC / MS[M+H]1173.4 (calculated value), LC / MS[M+H]1173.5 (observed value).

[0348] Example L-115 4-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[4-[2-amino-4- [Ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]-4-methyl-3,5-dioxo-1-piperidyl]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] Synthesis of ethoxy]ethoxy]ethoxy]ethoxy]propanoyloxy]-2,3,5,6-tetrafluoro-benzenesulfonic acid, HxBzL-115 [ka] Tert-Butyl 4-[4-[ethoxy(propyl)carbamoyl]-2-(tritylamino)-3H-1-benzazepin-8-yl]-3,5-dioxo-piperidine-1-carboxylate HxBzL-115b preparation 8-bromo-N-ethoxy-N-propyl-2-(tritylamino)-3H-1-benzazepine-4-carboxamide, HxBzL-115a (1 g, 1.64 mmol, 1.0 eq.) and tert- in dioxane (20 mL). Cs 2 C.O. 3 (1.07g, 3.29mmol, 2.0eq) and [2-(2-aminophenyl)phenyl]-chloro-palladium, dicyclohexyl-[3-(2,4,6-triisopropylphenyl)phenyl]phosphane, XPhos Pd G2 , CAS Reg.No.1310584-14-5 (129 mg, 164 umol, 0.1 equivalent), N 2was added in one portion at 25° C. and then stirred at 100° C. for 12 hours. The mixture was diluted with water and extracted with EtOAc (30 mL x 3). Wash the organic layer with brine and remove Na 2 S.O. 4 Dry on top, filter and concentrate. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, ethyl acetate / MeOH=1 / 0, 10 / 1) to obtain HxBzL-115b (0.35 g, 472.41 umol, yield 28.75%) as a yellow solid. LC / MS[M+H]741.36 (calculated value), LC / MS[M+H]741.4 (observed value).

[0349] Tert-Butyl 4-[4-[ethoxy(propyl)carbamoyl]-2-(tritylamino)-3H-1-benzazepin-8-yl]-4-methyl-3,5-dioxo-piperidine-1-carboxylate , Preparation of HxBzL-115c A mixture of HxBzL-115b (0.2 g, 269 umol, 1.0 equiv) in DMF (1 mL) was 2 C.O. 3 (74.6 mg, 539 umol, 2.0 eq) and MeI (11.5 mg, 80.9 umol, 5.04 uL, 0.3 eq) were added in one portion at 25°C and then stirred at 25°C for 12 hours. The mixture was diluted with water and extracted with EtOAc (20 mL x 3). Wash the organic layer with brine and remove Na 2 S.O. 4 Dry on top, filter and concentrate. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 1 / 1) to obtain HxBzL-115c (0.12 g, 158.96 umol, Yield 58.89%) was obtained as a yellow oil. LC / MS[M+H]755.37 (calculated value), LC / MS[M+H]755.4 (observed value).

[0350] Preparation of 2-amino-N-ethoxy-8-(4-methyl-3,5-dioxo-4-piperidyl)-N-propyl-3H-1-benzazepine-4-carboxamide, HxBzL-115d To a mixture of HxBzL-115c (0.12 g, 159 umol, 1.0 eq) in DCM (4 mL) was added TFA (362 mg, 3.18 mmol, 235 uL, 20.0 eq) in one portion at 25 °C and then at 50 °C for 12 Stir for hours. The mixture was concentrated in vacuo to give HxBzL-115d (0.07g, crude, TFA) as a yellow oil. LC / MS[M+H]413.2 (calculated value), LC / MS[M+H]413.2 (observed value).

[0351] 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[4-[2-amino-4-[ethoxy(propyl)carbamoyl]- 3H-1-Benzazepin-8-yl]-4-methyl-3.5-dioxo-1-piperidyl]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid , Preparation of HxBzL-115e A mixture of HxBzL-115d (30 mg, 56.9 umol, 1.0 eq., TFA) in DMF (1 mL) was added with Et3N (17.3 mg, 171 umol, 23.8 uL, 3.0 eq.) and 3-[2-[2-[2-[ 2-[2-[2-[2-[2-[2-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]ethoxy]ethoxy]ethoxy]propanoic acid, HOOC-PEG10-TFP (48.3 mg, 68.4 umol, 1.2 eq.) was added in one portion at 25°C and then stirred at 25°C for 0.5 h. The mixture was filtered and purified by preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 5%~35%, 8 min) to obtain HxBzL. -115e (7 mg, 7.34 umol, 12.89% yield) was obtained as a yellow oil. LC / MS[M+H]953.5 (calculated value), LC / MS[M+H]953.5 (observed value).

[0352] Preparation of HxBzL-115 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[3-[4-[2 -amino-4-[ethoxy(propyl)carbamoyl]-3H-1-benzazepin-8-yl]-4-methyl-3,5-dioxo-1-piperidyl]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy To a mixture of ]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (7 mg, 7.34 umol, 1.0 eq.) was added sodium 2,3,5,6-tetrafluoro-4-hydroxy-benzenesulfonate (7.88 mg , 29.4 mol, 4.0 eq) and EDCI (7.04 mg, 36.72 umol, 5.0 eq) were added in one portion at 25°C and then stirred at 25°C for 0.5 h. The mixture was filtered and preparative HPLC (column: Phenomenex Luna80*30mm*3um, mobile phase: [water (0.1%TFA)-ACN], B%: 15%~45%, 8 min column: Phenomenex Luna80*30mm* 3 um, mobile phase: [water (0.1% TFA)-ACN], B%: 15%~45%, 8 min) to produce HxBzL-115 (3 mg, 2.54 umol, yield 34.58%) as a yellow solid. obtained as. 1 H NMR (MeOD, 400 MHz) δ 7.59 (d, J = 8.0 Hz, 1H), 7.36 (s, 1H), 7.06-6.98 (m, 2H), 5.25 (dd, J=2.0, 19.6 Hz, 1H) , 4.14 (d, J = 19.6 Hz, 1H), 3.95 (q, J = 7.2 Hz, 2H), 3.87 (t, J = 6.0 Hz, 2H), 3.76-3.69 (m, 2H), 3.67-3.56 ( m, 36H), 3.55-3.50 (m, 2H), 3.45-3.35 (m, 4H), 2.98 (t, J = 6.0 Hz, 2H), 2.55-2.41 (m, 1H), 2.06-1.91 (m, 1H), 1.83-1.67 (m, 2H), 1.44 (s, 3H), 1.18 (t, J = 7.2 Hz, 3H), 0.98 (t, J = 7.2 Hz, 3H).LC / MS[M+H ]1181.4 (calculated value), LC / MS[M+H]1181.5 (observed value).

[0353] Example 201 Preparation of immune complex (IC) To prepare lysine conjugate immune complexes, antibodies were coated on a G-25 SEPHADEX™ desalting column (Sigma-Aldrich, St. Louis, MO) or on a Zeba™ spin desalting column (Thermo Fisher Scientific). Buffer exchange using a conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3. Each eluate is then adjusted to a concentration of approximately 1-10 mg / ml using buffer and then sterile filtered. The antibody was pre-warmed to 20-30°C and 2-20 (e.g. 7-10) molar equivalents of tetra of formula II dissolved in dimethylsulfoxide (DMSO) or dimethylacetamide (DMA) to a concentration of 5-20mM. Mix rapidly with fluorophenyl (TFP) or sulfonic acid tetrafluorophenyl (sulfoTFP) ester, 8-Het-2-aminobenzazepine-linker (HxBzL) compound. Reactions were allowed to proceed for approximately 16 h at 30 °C, and immune complexes (ICs) were equilibrated on two consecutive G-25 desalting columns or in phosphate-buffered saline (PBS) at pH 7.2. Separated from the reactions by running a Zeba™ spin desalting column to provide the immunoconjugates (IC) of Tables 3a and 3b. Adjuvant-to-antibody ratio (DAR) was determined using a C4 reversed phase column on an ACQUITY™ UPLC H class (Waters Corporation, Milford, MA) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Determined by liquid chromatography mass spectrometry used.

[0354] To prepare cysteine ​​conjugate immune complexes, antibodies were transferred using Zeba™ spin desalting columns (Thermo Fisher Scientific) to conjugation buffer containing PBS, pH 7.2 with 2 mM EDTA. Exchange the buffer. Reduce interchain disulfides using a 2-4 molar excess of tris(2-carboxyethyl)phosphine (TCEP) or dithiothreitol (DTT) for 30 min to 2 h at 37 °C. Excess TCEP or DTT was removed using a Zeba™ spin desalting column pre-equilibrated with conjugation buffer. The concentration of buffer exchanged antibodies was adjusted to approximately 5-20 mg / ml using conjugation buffer and sterile filtered. Maleimide-HxBzL compounds are dissolved in either dimethyl sulfoxide (DMSO) or dimethylacetamide (DMA) to a concentration of 5-20 mM. For conjugation, the antibody is mixed with 10-20 molar equivalents of maleimide-HxBzL. In some cases, up to 20% (v / v) additional DMA or DMSO was added to improve the solubility of maleimide-HxBzL in the conjugation buffer. The reaction is allowed to proceed at 20°C for approximately 30 minutes to 4 hours. The resulting conjugate is purified from unreacted maleimide-HxBzL using two consecutive Zeba™ spin desalting columns. The column is pre-equilibrated with phosphate buffered saline (PBS) at pH 7.2. Adjuvant-to-antibody ratio (DAR) was measured using a C4 reversed phase column on an ACQUITY™ UPLC H class (Waters Corporation, Milford, MA) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Estimated by liquid chromatography mass spectrometry used.

[0355] For conjugation, antibodies can be dissolved in aqueous buffer systems known in the art that do not adversely affect antibody stability or antigen binding specificity. Phosphate buffered saline may also be used. The HxBzL compound is dissolved in a solvent system that includes at least one polar aprotic solvent, as described elsewhere herein. In some such embodiments, HxBzL is in a pH 8 Tris buffer (e.g., 50 mM Tris) at about 5 mM, about 10 mM, about 20 mM, about 30 mM, about 40 mM, or about 50 mM, and from about 5 mM to about 50 mM, or It is dissolved in a concentration range such as about 10mM to about 30mM. In some embodiments, the 8-Het-2-aminobenzazepine-linker intermediate is dissolved in DMSO (dimethylsulfoxide), DMA (dimethylacetamide), acetonitrile, or another suitable dipolar aprotic solvent. Ru.

[0356] Alternatively, an equivalent excess of HxBzL solution may be diluted and combined with the antibody solution in a conjugation reaction. The HxBzL solution may be suitably diluted with at least one polar aprotic solvent and at least one polar protic solvent, examples of which include water, methanol, ethanol, n-propanol, and acetic acid. The molar equivalent of 8-Het-2-aminobenzazepine-linker intermediate to antibody is about 1.5:1, about 3:1, about 5:1, about 10:1, about 15:1, or about 20:1. 1, and about 1.5:1 to about 20:1, about 1.5:1 to about 15:1, about 1.5:1 to about 10:1, about 3:1 to about 15:1, about 3:1 to about 10:1, about 5:1 to about 15:1, or about 5:1 to about 10:1. The reaction may be suitably monitored for completion by methods known in the art such as LC-MS. The conjugation reaction is typically complete in a range of about 1 hour to about 16 hours. After the reaction is complete, reagents may be added to the reaction mixture to quench the reaction. If the antibody thiol group has been reacted with a thiol-reactive group such as the maleimide of the 8-Het-2-aminobenzazepine-linker intermediate, the unreacted antibody thiol group may be reacted with a capping reagent. . An example of a suitable capping reagent is ethylmaleimide.

[0357] After conjugation, the immune complex can be purified by chromatography, such as size exclusion chromatography, hydrophobic interaction chromatography, ion exchange chromatography, chromatofocusing, ultrafiltration, centrifugal ultrafiltration, tangential flow filtration, and combinations thereof. may be purified and separated from unconjugated reactants and / or conjugate aggregates by purification methods known in the art. For example, purification may be preceded by diluting the immune complex, such as in 20 mM sodium succinate, pH 5. The diluted solution is applied to the cation exchange column, followed by washing with, for example, at least 10 column volumes of 20 mM sodium succinate, pH 5. The conjugate may be suitably eluted with a buffer such as PBS.

[0358] Example 202 HEK reporter assay HEK293 reporter cells expressing human TLR7 or human TLR8 were purchased from Invivogen, and cell growth and experiments were performed according to vendor protocols. Briefly, cells were incubated with 5% CO in DMEM supplemented with 10% FBS, Zeocin, and Blasticidin. 2 and grown to 80-85% fusion. Then, using HEK detection medium and substrate containing immunostimulatory molecules, 4 x 10 4 Cells were seeded in 96-well plates at cells / well. Activity was measured using a plate reader at wavelengths of 620-655 nm.

[0359] Example 203 Evaluation of in vivo immune complex activity This example shows that the immunoconjugates of the invention are effective in inducing immune activation and are therefore useful in the treatment of cancer.

[0360] a) Isolation of human antigen-presenting cells: Human bone marrow antigen-presenting cells (APCs) were isolated from the ROSETTESEP™ human monocyte enrichment cocktail containing monoclonal antibodies against CD14, CD16, CD40, CD86, CD123, and HLA-DR. Negative selection was made from human peripheral blood obtained from healthy blood donors (Stanford Blood Center, Palo Alto, California) by density gradient centrifugation using Stem Cell Technologies, Vancouver, Canada). Immature APCs were then harvested using the EASYSEP™ Human Monocyte Enrichment Kit without CD16 Depletion (Stem Cell Technologies) containing monoclonal antibodies against CD14, CD16, CD40, CD86, CD123, and HLA-DR. , purified to >90% purity by negative selection.

[0361] b) Myeloid APC activation assay: 2x10 5 APC was cultured in Iscove's modified dulbecco's medium, IMDM (Lonza) supplemented with 10% FBS, 100 U / mL penicillin, 100 μg / mL (micrograms per milliliter) streptomycin, 2 mM L-glutamine, sodium pyruvate, non-essential amino acids, and as indicated. 96-well plates (Corning, Corning, NY) containing various concentrations of unconjugated (naked) antibodies of the invention and immunoconjugates (prepared according to the Examples above). Cell-free supernatants are analyzed after 18 hours via ELISA to measure TNFα secretion as a readout of the pro-inflammatory response.

[0362] c) PBMC activation assay: Human peripheral blood mononuclear cells were isolated from human peripheral blood obtained from healthy blood donors (Stanford Blood Center, Palo Alto, California) by density gradient centrifugation. PBMCs were incubated in 96-well plates (Corning, Corning, NY) co-cultured with CEA-expressing tumor cells (eg, MKN-45, HPAF-II) at a 10:1 effector to target cell ratio. Cells were stimulated with various concentrations of unconjugat...

Claims

1. 1. The method of claim 1, further comprising administering to said patient an antibody having formula I: Ab-[L-HxBz] ｐ I An immune complex having the formula: During the ceremony, Ab is an antibody construct having an antigen-binding domain that binds to HER2; p is an integer from 1 to 8; HxBz is a compound of the formula: 【Chemistry 1】 wherein Het is selected from heterocyclyldiyl and heteroaryldiyl; R １ , R ２ , R ３ , and R ４ However, independently, H, C １ -C １２ Alkyl, C ２ -C ６ Alkenyl, C ２ -C ６ Alkynyl, C ３ -C １２ Carbocyclyl, C ６ -C ２０ Aryl, C ２ -C ９ Heterocyclyl, and C １ -C ２０ heteroaryl, where alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heterocyclyl, and heteroaryl are independently and optionally selected from the group consisting of: - (C １ -C １２ alkyldiyl)-N(R ５ ) - *, - (C １ -C １２ alkyldiyl)-N(R ５ ) ２ , - (C １ -C １２ Alkyldiyl)-OR ５ , - (C ３ -C １２ Carbocyclyl), - (C ３ -C １２ Carbocyclyl)-*, - (C ３ -C １２ carbocyclyl)-(C １ -C １２ Alkyldiyl)-NR ５ - *, - (C ３ -C １２ carbocyclyl)-(C １ -C １２ alkyldiyl)-N(R ５ ) ２ , - (C ３ -C １２ carbocyclyl)-NR ５ -C(=NR ５ ) N.R. ５ - *, - (C ６ -C ２０ Aryl), - (C ６ -C ２０ aryl)-*, - (C ６ -C ２０ Aryldiyl)-N(R ５ ) - *, - (C ６ -C ２０ Aryldiyl)-(C １ -C １２ alkyldiyl)-N(R ５ ) - *, - (C ６ -C ２０ Aryldiyl)-(C １ -C １２ Alkyldiyl)-(C ２ -C ２０ Heterocyclyldiyl)-*, - (C ６ -C ２０ Aryldiyl)-(C １ -C １２ alkyldiyl)-N(R ５ ) ２ , - (C ６ -C ２０ Aryldiyl)-(C １ -C １２ Alkyldiyl)-NR ５ -C(=NR ５ａ ) N (R ５ ) - *, - (C ２ -C ２０ Heterocyclyl), - (C ２ -C ２０ Heterocyclyl)-*, - (C ２ -C ９ Heterocyclyl)-(C １ -C １２ Alkyldiyl)-NR ５ - *, - (C ２ -C ９ Heterocyclyl)-(C １ -C １２ alkyldiyl)-N(R ５ ) ２ , - (C ２ -C ９ Heterocyclyl)-C(=O)-(C １ -C １２ alkyldiyl)-N(R ５ ) - *, - (C ２ -C ９ Heterocyclyl)-NR ５ -C(=NR ５ａ ) N.R. ５ - *, - (C ２ -C ９ Heterocyclyl)-NR ５ - (C ６ -C ２０ Aryldiyl)-(C １ -C １２ alkyldiyl)-N(R ５ ) - *, - (C ２ -C ９ Heterocyclyl)-(C ６ -C ２０ Aryldiyl)-*, - (C １ -C ２０ Heteroaryl), - (C １ -C ２０ Heteroaryl)-*, - (C １ -C ２０ Heteroaryl)-(C １ -C １２ alkyldiyl)-N(R ５ ) - *, - (C １ -C ２０ Heteroaryl)-(C １ -C １２ alkyldiyl)-N(R ５ ) ２ , - (C １ -C ２０ Heteroaryl)-NR ５ -C(=NR ５ａ ) N (R ５ ) - *, - (C １ -C ２０ Heteroaryl)-N(R ５ ) C(=O)-(C １ -C １２ alkyldiyl)-N(R ５ ) - *, -C(=O)-*, -C(=O)-(C １ -C １２ alkyldiyl)-N(R ５ ) - *, -C(=O)-(C ２ -C ２０ Heterocyclyldiyl)-*, -C(=O)N(R ５ ) ２ 、 -C(=O)N(R ５ )-*、 -C(=O)N(R ５ )-(C １ -C １２ alkyldiyl)-N(R ５ ) C(=O)R ５ , -C(=O)N(R ５ )-(C １ -C １２ alkyldiyl)-N(R ５ )C(=O)N(R ５ ) ２ , -C(=O)NR ５ - (C １ -C １２ alkyldiyl)-N(R ５ ) CO ２ R ５ , -C(=O)NR ５ - (C １ -C １２ alkyldiyl)-N(R ５ ) C(=NR ５ａ ) N (R ５ ) ２ , -C(=O)NR ５ - (C １ -C １２ Alkyldiyl)-NR ５ C (=NR ５ａ ) R ５ , -C(=O)NR ５ - (C １ -C ８ Alkyldiyl)-NR ５ (C ２ -C ５ Heteroaryl), -C(=O)NR ５ - (C １ -C ２０ Heteroaryldiyl)-N(R ５ ) - *, -C(=O)NR ５ - (C １ -C ２０ Heteroaryldiyl)-*, -C(=O)NR ５ - (C １ -C ２０ Heteroaryldiyl)-(C １ -C １２ alkyldiyl)-N(R ５ ) ２ , -C(=O)NR ５ - (C １ -C ２０ Heteroaryldiyl)-(C ２ -C ２０ Heterocyclyldiyl)-C(=O)NR ５ - (C １ -C １２ Alkyldiyl)-NR ５ - *, -N(R ５ ) ２ 、 -N(R ５ )-*、 -N(R ５ )C(=O)R ５ 、 -N(R ５ )C(=O)-*、 -N(R ５ )C(=O)N(R ５ ) ２ 、 -N(R ５ )C(=O)N(R ５ )-*、 -N(R ５ )CO ２ R ５ 、 -NR ５ C(=NR) ５ａ )N(R ５ ) ２ 、 -NR ５ C(=NR) ５ａ )N(R ５ )-*、 -NR ５ C(=NR) ５ａ )R ５ 、 -N(R ５ ) C(=O)-(C １ -C １２ alkyldiyl)-N(R ５ ) - *, -N(R ５ )-(C ２ -C ５ Heteroaryl), -N(R ５ ) -S(=O) ２ - (C １ -C １２ alkyl), -O-(C １ -C １２ alkyl), -O-(C １ -C １２ alkyldiyl)-N(R ５ ) ２ , -O-(C １ -C １２ alkyldiyl)-N(R ５ ) - *, -O-C(=O)N(R ５ ) ２ 、 -O-C(=O)N(R ５ )-*、 -S(=O) ２ - (C ２ -C ２０ Heterocyclyldiyl)-*, -S(=O) ２ - (C ２ -C ２０ Heterocyclyldiyl)-(C １ -C １２ alkyldiyl)-N(R ５ ) ２ , -S(=O) ２ - (C ２ -C ２０ Heterocyclyldiyl)-(C １ -C １２ Alkyldiyl)-NR ５ -*, and -S(=O) ２ - (C ２ -C ２０ Heterocyclyldiyl)-(C １ -C １２ or substituted with one or more groups selected from: Or R ２ and R ３ together form a 5- or 6-membered heterocyclyl ring, X １ , X ２ , X ３ , and X ４ are independently a bond, C(=O), C(=O)N(R ５ ), O, N(R ５ ), S, S(O) ２ , and S(O) ２ N (R ５ ) selected from the group consisting of R ５ However, independently, H, C ６ -C ２０ Aryl, C ３ -C １２ Carbocyclyl, C ６ -C ２０ Aryldiyl, C １ -C １２ Alkyl, and C １ -C １２ alkyldiyl, or two R ５ the groups taken together form a 5- or 6-membered heterocyclyl ring; R ５ａ But, C ６ -C ２０ Aryl and C １ -C ２０ Heteroaryl; In the formula, the asterisk * indicates the binding site of L, and R １ , R ２ , R ３ , and R ４ is attached to L; L, -C(=O)-PEG-, -C(=O)-PEG-C(=O)N(R ６ )-(C １ -C １２ alkyldiyl)-C(=O)-Gluc-, -C(=O)-PEG-O-, -C(=O)-PEG-O-C(=O)-, -C(=O)-PEG-C(=O)-, -C(=O)-PEG-C(=O)-PEP-, -C(=O)-PEG-N(R ６ )-、 -C(=O)-PEG-N(R ６ )-C(=O)-、 -C(=O)-PEG-N(R ６ )-PEG-C(=O)-PEP-、 -C(=O)-PEG-N ＋ (R ６ ) ２ 2019-02-20 10:00:00 AM -C(=O)-PEG-C(=O)-PEP-N(R ６ )-(C １ -C １２ Alkyldiyl)-, -C(=O)-PEG-C(=O)-PEP-N(R ６ )-(C １ -C １２ Alkyldiyl)N(R ６ ) C(=O)-(C ２ -C ５ Monoheterocyclyldiyl)-, -C(=O)-PEG-SS-(C １ -C １２ alkyldiyl)-OC(=O)-, -C(=O)-PEG-SS-(C １ -C １２ alkyldiyl)-C(=O)-, -C(=O)-(C １ -C １２ alkyldiyl)-C(=O)-PEP-, -C(=O)-(C １ -C １２ alkyldiyl)-C(=O)-PEP-N(R ６ )-(C １ -C １２ Alkyldiyl)-, -C(=O)-(C １ -C １２ alkyldiyl)-C(=O)-PEP-N(R ６ )-(C １ -C １２ alkyldiyl)-N(R ５ )-C(=O), -C(=O)-(C １ -C １２ alkyldiyl)-C(=O)-PEP-N(R ６ )-(C １ -C １２ alkyldiyl)-N(R ６ ) C(=O)-(C ２ -C ５ Monoheterocyclyldiyl)-, -Succinimidyl-(CH ２ ) ｍ -C(=O)N(R ６ )-PEG-, -Succinimidyl-(CH ２ ) ｍ -C(=O)N(R ６ )-PEG-C(=O)N(R ６ )-(C １ -C １２ alkyldiyl)-C(=O)-Gluc-, -Succinimidyl-(CH ２ ) ｍ -C(=O)N(R ６ )-PEG-O-, -Succinimidyl-(CH ２ ) ｍ -C(=O)N(R ６ )-PEG-O-C(=O)-, -Succinimidyl-(CH ２ ) ｍ -C(=O)N(R ６ )-PEG-C(=O)-, -Succinimidyl-(CH ２ ) ｍ -C(=O)N(R ６ )-PEG-N(R ５ ) -, -Succinimidyl-(CH ２ ) ｍ -C(=O)N(R ６ )-PEG-N(R ５ )-C(=O)-, -Succinimidyl-(CH ２ ) ｍ -C(=O)N(R ６ )-PEG-C(=O)-PEP-, -Succinimidyl-(CH ２ ) ｍ -C(=O)N(R ６ )-PEG-SS-(C １ -C １２ alkyldiyl)-OC(=O)-, -Succinimidyl-(CH ２ ) ｍ -C(=O)-PEP-N(R ６ )-(C １ -C １２ Alkyldiyl)-, -Succinimidyl-(CH ２ ) ｍ -C(=O)-PEP-N(R ６ )-(C １ -C １２ Alkyldiyl)N(R ６ ) C(═O)—, and -Succinimidyl-(CH ２ ) ｍ -C(=O)-PEP-N(R ６ )-(C １ -C １２ Alkyldiyl)N(R ６ ) C(=O)-(C ２ -C ５ the linker being selected from the group consisting of: R ６ are independently H or C １ -C ６ is alkyl, PEG is a group represented by the formula: -(CH ２ CH ２ O) ｎ - (CH ２ ) ｍ -, m is an integer from 1 to 5, and n is an integer from 2 to 50; Gluc has the formula: 【Chemistry 2】 having PEP has the formula: 【Chemistry 3】 where AA are independently selected from natural or unnatural amino acid side chains, or one or more of AA and the adjacent nitrogen atom form a 5-membered ring proline amino acid, and the wavy line indicates the point of attachment; Cyc is optionally F, Cl, NO ２ , -OH, -OCH ３ and glucuronic acid having the structure: ６ -C ２０ Aryldiyl and C １ -C ２０ heteroaryldiyl; 【Chemistry 4】 R ７ is -CH(R ８ ) O-, -CH ２ --, --CH ２ N (R ８ )- and -CH(R ８ )O—C(═O)—, wherein R ８ But, H, C １ -C ６ Alkyl, C(=O)-C １ -C ６ Alkyl, and -C(=O)N(R ９ ) ２ wherein R ９ However, independently, H, C １ -C １２ Alkyl, and -(CH ２ CH ２ O) ｎ - (CH ２ ) ｍ -OH, wherein m is an integer from 1 to 5 and n is an integer from 2 to 50, or two R ９ the groups taken together form a 5- or 6-membered heterocyclyl ring; y is an integer from 2 to 12; z is 0 or 1; Alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl are independently and optionally selected from F, Cl, Br, I, -CN, -CH ３ , -CH ２ CH ３ , -CH=CH ２ , -C≡CH, -C≡CCH ３ , -CH ２ CH ２ CH ３ , -CH(CH ３ ) ２ , -CH ２ CH (CH ３ ) ２ , -CH ２ OH, -CH ２ OCH ３ , -CH ２ CH ２ OH, -C(CH ３ ) ２ OH, -CH(OH)CH(CH ３ ) ２ , -C(CH ３ ) ２ CH ２ OH, -CH ２ CH ２ SO ２ CH ３ , -CH ２ OP(O)(OH) ２ , -CH ２ F, -CHF ２ , -CF ３ , -CH ２ CF ３ , -CH ２ CHF ２ , -CH(CH ３ )CN, -C(CH ３ ) ２ CN, -CH ２ CN, -CH ２ N.H. ２ , -CH ２ N.H.S.O. ２ CH ３ , -CH ２ N.H.C.H. ３ , -CH ２ N(CH ３ ) ２ 、-CO ２ H、-COCH ３ 、-CO ２ CH ３ 、-CO ２ C(CH ３ ) ３ 、-COCH(OH)CH ３ 、-CONH ２ 、-CONHCH ３ 、-CON(CH ３ ) ２ 、-C(CH ３ ) ２ P. ２ 、-NH ２ 、-NHCH ３ 、-N(CH ３ ) ２ -NHCOCH ３ 、-N(CH ３ )COCH ３ 、-NHS(O) ２ CH ３ 、-N(CH ３ )C(CH ３ ) ２ P. ２ 、-N(CH ３ )CH ２ CH ２ S(O) ２ CH ３ 、-NHC(=NH)H, -NHC(=NH)CH ３ -NHC(=NH)NH ２ -NHC(=O)NH ２ 、-NO ２ 、=O、-OH、-OCH ３ 、-OCH ２ CH ３ 、-OCH ２ CH ２ SO ３ 、-OCH ２ CH ２ H ２ CH ２ N(CH ３ ) ２ 、-O(CH ２ CH ２ O) ｎ -(CH ２ ) ｍ CO ２ H, -O(CH ２ CH ２ O) ｎ H, -OCH ２ F, -OCHF ２ , -OCF ３ , -OP(O)(OH) ２ , -S(O) ２ N (CH ３ ) ２ , -SCH ３ , -S(O) ２ CH ３ , and -S(O) ３ H, or a pharma- ceutically acceptable salt thereof.

2. The immunoconjugate of claim 1 , wherein the antibody is selected from trastuzumab and pertuzumab.

3. 3. The immunoconjugate of claim 1, wherein Het is selected from the group consisting of pyridyldiyl, pyrimidyldiyl, pyrazolyldiyl, piperazinyldiyl, piperidinyldiyl, and pyrazinyldiyl.

4. X １ is a bond, and R １ The immune complex according to any one of claims 1 and 2, wherein is H.

5. X ２ is a bond, and R ２ But, C １ -C ８ The immunoconjugate of claim 1 or 2, which is alkyl.

6. X ２ and X ３ are each a bond, R ２ and R ３ But independently, C １ -C ８ Alkyl, —O—(C １ -C １２ alkyl), -(C １ -C １２ Alkyldiyl)-OR ５ , -(C １ -C ８ alkyldiyl)-N(R ５ ) CO ２ R ５ , -(C １ -C １２ alkyl)-OC(O)N(R ５ ) ２ , -O-(C １ -C １２ alkyl)-N(R ５ ) CO ２ R ５ , and —O—(C １ -C １２ alkyl)-OC(O)N(R ５ ) ２ The immune complex according to claim 1 or 2, which is selected from the group consisting of

7. R ２ But, C １ -C ８ alkyl, R ３ However, (C １ -C ８ alkyldiyl)-N(R ５ ) CO ２ R ５ The immune complex of claim 6,

8. R ２ But -CH ２ CH ２ CH ３ and R ３ But -CH ２ CH ２ CH ２ N.H.C.O. ２ (t-Bu), -OCH ２ CH ２ N.H.C.O. ２ (cyclobutyl), and -CH ２ CH ２ CH ２ N.H.C.O. ２ 8. The immune conjugate of claim 7, wherein said aryl group is selected from the group consisting of aryl, ... and aryl.

9. R ２ and R ３ each independently represents -CH ２ CH ２ CH ３ , -OCH ２ CH ３ , -OCH ２ CF ３ , -CH ２ CH ２ CF ３ , -OCH ２ CH ２ OH and -CH ２ CH ２ CH ２ 7. The immune complex of claim 6, wherein said amino acid is selected from the group consisting of OH.

10. R ２ and R ３ are each -CH ２ CH ２ CH ３ The immune complex of claim 9,

11. R ２ But -CH ２ CH ２ CH ３ and R ３ But, -OCH ２ CH ３ The immune complex of claim 6,

12. X ３ -R ３ but, 【Chemistry 5】 The immune complex according to any one of claims 1 and 2, selected from the group consisting of:

13. X ４ is a bond, and R ４ The immune complex according to any one of claims 1 and 2, wherein is H.

14. R １ The immune complex according to claim 1 or 2, wherein:

15. R ２ Or R ３ The immune complex according to claim 1 or 2, wherein:

16. X ３ -R ３ -L is, 【Chemistry 6】 16. The immunoconjugate of claim 15, wherein the wavy line indicates the point of attachment to N.

17. The immunoconjugate according to any one of claims 1 and 2, wherein L is -C(=O)-PEG- or -C(=O)-PEG-C(=O)-.

18. 3. The immune complex of claim 1, wherein L is attached to a cysteine ​​thiol of the antibody.

19. The immunoconjugate of any one of claims 1 and 2, wherein for the PEG, m is 1 or 2 and n is an integer from 2 to 10.

20. 20. The immune complex of claim 19, wherein n is 10.

21. L has the structure: 【Chemistry 12】 wherein the wavy line is selected from R ５ 3. The immune complex of claim 1, which exhibits binding to

22. Formula Ia: 【Chemistry 13】 The immune complex according to claim 1 , having the formula:

23. X ４ is a bond, and R ４ The immune complex of claim 22, wherein is H.

24. X ２ and X ３ are each a bond, R ２ and R ３ But independently, C １ -C ８ Alkyl, —O—(C １ -C １２ alkyl), -(C １ -C １２ Alkyldiyl)-OR ５ , -(C １ -C ８ alkyldiyl)-N(R ５ ) CO ２ R ５ , -(C １ -C １２ alkyl)-OC(O)N(R ５ ) ２ , -O-(C １ -C １２ alkyl)-N(R ５ ) CO ２ R ５ , and —O—(C １ -C １２ alkyl)-OC(O)N(R ５ ) ２ 23. The immune complex of claim 22, selected from:

25. Formulae Ib to Ii: 【Chemistry 14-1】 【Chemistry 14-2】 23. The immune complex of claim 22, selected from:

26. X ２ and X ３ are each a bond, R ２ and R ３ But independently, C １ -C ８ Alkyl, —O—(C １ -C １２ alkyl), -(C １ -C １２ Alkyldiyl)-OR ５ , -(C １ -C ８ alkyldiyl)-N(R ５ ) CO ２ R ５ , and —O—(C １ -C １２ alkyl)-N(R ５ ) CO ２ R ５ 26. The immune complex of claim 25, selected from:

27. X ２ and X ３ are each a bond, R ２ But, C １ -C ８ alkyl, R ３ But -O-(C １ -C １２ alkyl) and -O-(C １ -C １２ alkyl)-N(R ５ ) CO ２ R ５ 26. The immune complex of claim 25, selected from:

28. An 8-Het-2-aminobenzazepine-linker compound selected from the group consisting of:

29. 29. An immunoconjugate prepared by conjugation of an anti-HER2 antibody with an 8-Het-2-aminobenzazepine-linker compound according to claim 28.

30. A pharmaceutical composition comprising a therapeutically effective amount of the immunoconjugate of any one of claims 1 and 2, and one or more pharma- ceutically acceptable diluents, vehicles, carriers, or excipients.

31. A pharmaceutical for treating cancer comprising the immunoconjugate described in any one of claims 1 to 27, wherein the cancer is selected from cervical cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, esophageal cancer, bladder cancer, urinary tract cancer, urothelial cancer, lung cancer, non-small cell lung cancer, Merkel cell carcinoma, colon cancer, colorectal cancer, gastric cancer, and breast cancer.

32. A method for preparing an immunoconjugate of formula I according to claim 1, wherein the 8-Het-2-amino-thienoazepine-linker compound according to claim 28 is conjugated to said anti-HER2 antibody.

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