Patent Publication Number: US-2022226492-A1

Title: Immunoconjugates Targeting HER2

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This patent application claims the benefit of U.S. Provisional Patent Application No. 62/819,345, filed Mar. 15, 2019, which is incorporated by reference in its entirety herein. 
    
    
     INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY 
     Incorporated by reference in its entirety herein is a computer-readable nucleotide/amino acid sequence listing submitted concurrently herewith and identified as follows: one 28,602 Byte ASCII (Text) file named “748782_ST25.txt,” created Mar. 9, 2020. 
     BACKGROUND OF THE INVENTION 
     It is now well appreciated that tumor growth necessitates the acquisition of mutations that facilitate immune evasion. Even so, tumorigenesis results in the accumulation of mutated antigens, or neoantigens, that are readily recognized by the host immune system following ex vivo stimulation. Why and how the immune system fails to recognize neoantigens are beginning to be elucidated. Groundbreaking studies by Carmi et al. ( Nature,  521: 99-104 (2015)) have indicated that immune ignorance can be overcome by delivering neoantigens to activated dendritic cells via antibody-tumor immune complexes. In these studies, simultaneous delivery of tumor binding antibodies and dendritic cell adjuvants via intratumoral injections resulted in robust anti-tumor immunity. New compositions and methods for the delivery of antibodies and dendritic cell adjuvants are needed in order to reach inaccessible tumors and/or to expand treatment options for cancer patients and other subjects. The invention provides such compositions and methods. 
     BRIEF SUMMARY OF THE INVENTION 
     The invention provides an immunoconjugate of formula (I): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 1  is CH or N, 
     J 2  is CHQ, NQ, O, or S, 
     each Q independently is Y or Z, wherein exactly one Q is Y, 
     Y is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     The invention provides an immunoconjugate of formula (II): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     each Q independently is Y or Z, wherein exactly one Q is Y, 
     Y is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     The invention provides a composition comprising a plurality of immunoconjugates described herein. 
     The invention provides a method for treating cancer in a subject comprising administering a therapeutically effective amount of an immunoconjugate or a composition described herein to a subject in need thereof. 
    
    
     
       BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS 
         FIG. 1A  shows the effect of Immunoconjugate A on myeloid activation in myeloid APC-tumor co-cultures, using the HCC1954 human ductal carcinoma tumor cell line. Median fluorescence intensity of co-stimulatory molecule CD40 (cells gated on viable CD45+CD11c+HLA-DR+) was measured by flow cytometry and is shown for trastuzumab (dotted line, circle), trastuzumab+Compound 7 (dashed line, triangle) or Immunoconjugate A (solid line, square). 
         FIG. 1B  shows the effect of Immunoconjugate A on myeloid activation in myeloid APC-tumor co-cultures, using the HCC1954 human ductal carcinoma tumor cell line. Median fluorescence intensity of co-stimulatory molecule CD86 (cells gated on viable CD45+CD11c+HLA-DR+) was measured by flow cytometry and is shown for trastuzumab (dotted line, circle), trastuzumab+Compound 7 (dashed line, triangle) or Immunoconjugate A (solid line, square). 
         FIG. 1C  shows the effect of Immunoconjugate A on myeloid activation in myeloid APC-tumor co-cultures, using the HCC1954 human ductal carcinoma tumor cell. TNFα secretion was measured by cytokine bead array (cells gated on viable CD45+CD11c+HLA-DR+) for trastuzumab (dotted line, circle), trastuzumab+Compound 7 (dashed line, triangle) or Immunoconjugate A (solid line, square). 
         FIG. 1D  shows the effect of Immunoconjugate A on myeloid activation in myeloid APC-tumor co-cultures, using the JIMT-1 human ductal carcinoma tumor cell line. Median fluorescence intensity of co-stimulatory molecule CD40 (cells gated on viable CD45+CD11c+HLA-DR+) was measured by flow cytometry and is shown for trastuzumab (dotted line, circle), trastuzumab+Compound 7 (dashed line, triangle) or Immunoconjugate A (solid line, square). 
         FIG. 1E  shows the effect of Immunoconjugate A on myeloid activation in myeloid APC-tumor co-cultures, using the JIMT-1 human ductal carcinoma tumor cell line. Median fluorescence intensity of co-stimulatory molecule CD86 (cells gated on viable CD45+CD11c+HLA-DR+) was measured by flow cytometry and is shown for trastuzumab (dotted line, circle), trastuzumab+Compound 7 (dashed line, triangle) or Immunoconjugate A (solid line, square). 
         FIG. 1F  shows the effect of Immunoconjugate A on myeloid activation in myeloid APC-tumor co-cultures, using the JIMT-1 human ductal carcinoma tumor cell. TNFα secretion was measured by cytokine bead array (cells gated on viable CD45+CD11c+HLA-DR+) for trastuzumab (dotted line, circle), trastuzumab+Compound 7 (dashed line, triangle) or Immunoconjugate A (solid line, square). 
         FIG. 1G  shows the effect of Immunoconjugate A on myeloid activation in myeloid APC-tumor co-cultures, using the COLO 205 human colon adenocarcinoma cell line. Median fluorescence intensity of co-stimulatory molecule CD40 (cells gated on viable CD45+CD11c+HLA-DR+) was measured by flow cytometry and is shown for trastuzumab (dotted line, circle), trastuzumab+Compound 7 (dashed line, triangle) or Immunoconjugate A (solid line, square). 
         FIG. 1H  shows the effect of Immunoconjugate A on myeloid activation in myeloid APC-tumor co-cultures, using the COLO 205 human colon adenocarcinoma cell line. Median fluorescence intensity of co-stimulatory molecule CD86 (cells gated on viable CD45+CD11c+HLA-DR+) was measured by flow cytometry and is shown for trastuzumab (dotted line, circle), trastuzumab+Compound 7 (dashed line, triangle) or Immunoconjugate A (solid line, square). 
         FIG. 1I  shows the effect of Immunoconjugate A on myeloid activation in myeloid APC-tumor co-cultures, using the COLO 205 human colon adenocarcinoma cell line. TNFα secretion was measured by cytokine bead array (cells gated on viable CD45+CD11c+HLA-DR+) for trastuzumab (dotted line, circle), trastuzumab+Compound 7 (dashed line, triangle) or Immunoconjugate A (solid line, square). 
         FIG. 2A  shows that Immunoconjugate B elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 2B  shows that Immunoconjugate B elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 2C  shows that Immunoconjugate B elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 2D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate B. 
         FIG. 3A  shows that Immunoconjugate C elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 3B  shows that Immunoconjugate C elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 3C  shows that Immunoconjugate C elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 3D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate C. 
         FIG. 4A  shows that Immunoconjugate D elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 4B  shows that Immunoconjugate D elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 4C  shows that Immunoconjugate D elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 4D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate D. 
         FIG. 5A  shows that Immunoconjugate E elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 5B  shows that Immunoconjugate E elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 5C  shows that Immunoconjugate E elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 5D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate E. 
         FIG. 6A  shows that Immunoconjugate F elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 6B  shows that Immunoconjugate F elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 6C  shows that Immunoconjugate F elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 6D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate F. 
         FIG. 6E  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate F. 
         FIG. 7A  shows that Immunoconjugate G elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 7B  shows that Immunoconjugate G elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 7C  shows that Immunoconjugate G elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 7D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate G. 
         FIG. 8A  shows that Immunoconjugate H elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 8B  shows that Immunoconjugate H elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 8C  shows that Immunoconjugate H elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 8D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate H. 
         FIG. 9A  shows that Immunoconjugate I elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 9B  shows that Immunoconjugate I elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 9C  shows that Immunoconjugate I elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 9D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate I. 
         FIG. 10A  shows that Immunoconjugate K1 elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 10B  shows that Immunoconjugate K1 elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 10C  shows that Immunoconjugate K1 elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 10D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate K1. 
         FIG. 11A  shows that Immunoconjugate L elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 11B  shows that Immunoconjugate L elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 11C  shows that Immunoconjugate L elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 12A  shows that Immunoconjugate M elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 12B  shows that Immunoconjugate M elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 12C  shows that Immunoconjugate M elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 12D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate M. 
         FIG. 13A  shows that Immunoconjugate N elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 13B  shows that Immunoconjugate N elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 14A  shows that Immunoconjugate O elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 14B  shows that Immunoconjugate O elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 14C  shows that Immunoconjugate O elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 14D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate O. 
         FIG. 15A  shows that Immunoconjugate P elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 15B  shows that Immunoconjugate P elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 15C  shows that Immunoconjugate P elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 15D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate P. 
         FIG. 16A  shows that Immunoconjugate Q elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 16B  shows that Immunoconjugate Q elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 17  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate R. 
         FIG. 18  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate R1. 
         FIG. 19A  shows that Immunoconjugate S elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 19B  shows that Immunoconjugate S elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 19C  shows that Immunoconjugate S elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 19D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate S. 
         FIG. 20A  shows that Immunoconjugate T elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 20B  shows that Immunoconjugate T elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 20C  shows that Immunoconjugate T elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 20D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate T. 
         FIG. 21  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate U. 
         FIG. 22A  shows that Immunoconjugate V elicits myeloid differentiation as indicated by CD14 downregulation. 
         FIG. 22B  shows that Immunoconjugate V elicits myeloid activation as indicated by CD40 upregulation. 
         FIG. 22C  shows that Immunoconjugate V elicits myeloid activation as indicated by CD86 upregulation. 
         FIG. 22D  shows TNFα secretion from myeloid cells following an 18 hour incubation with Immunoconjugate V. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The invention provides an immunoconjugate of formula (I) or (II). Antibody-adjuvant immunoconjugates of the invention, comprising an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”) linked to one or more adjuvants, demonstrate superior pharmacological properties over conventional antibody conjugates. The adjuvant/linker combinations are preferred to provide adequate purification and isolation of the immunoconjugate, maintain function of the one or more adjuvant moieties and antibody construct, and produce ideal pharmacokinetic (“PK”) properties of the immunoconjugate. Additional embodiments and benefits of the inventive antibody-adjuvant immunoconjugates will be apparent from description herein. 
     Definitions 
     As used herein, the term “immunoconjugate” refers to an antibody construct that is covalently bonded to an adjuvant moiety via a linker. 
     As used herein, the phrase “antibody construct” refers to an antibody or a fusion protein comprising (i) an antigen binding domain and (ii) an Fc domain. 
     As used herein, the term “antibody” refers to a polypeptide comprising an antigen binding region (including the complementarity determining region (CDRs)) from an immunoglobulin gene or fragments thereof that specifically binds and recognizes HER2. 
     An exemplary immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one “light” (about 25 kDa) and one “heavy” chain (about 50-70 kDa) connected by disulfide bonds. Each chain is composed of structural domains, which are referred to as immunoglobulin domains. These domains are classified into different categories by size and function, e.g., variable domains or regions on the light and heavy chains (V L  and V H , respectively) and constant domains or regions on the light and heavy chains (C L  and C H , respectively). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids, referred to as the paratope, primarily responsible for antigen recognition, i.e., 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 in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. IgG antibodies are large molecules of about 150 kDa composed of four peptide chains. IgG antibodies contain two identical class γ 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 and to a light chain each by disulfide bonds. The resulting tetramer has two identical halves, which together form the Y-like shape. Each end of the fork contains an identical antigen binding domain. There are four IgG subclasses (IgG1, IgG2, IgG3, and IgG4) in humans, named in order of their abundance in serum (i.e., IgG1 is the most abundant). Typically, the antigen binding domain of an antibody will be most critical in specificity and affinity of binding to cancer cells. 
     Antibodies can exist as intact immunoglobulins or as a number of well-characterized fragments produced by digestion with various peptidases. Thus, for example, pepsin digests an antibody below the disulfide linkages in the hinge region to produce F(ab)′ 2 , a dimer of Fab which itself is a light chain joined to V H -C H 1 by a disulfide bond. The F(ab)′ 2  may be reduced under mild conditions to break the disulfide linkage in the hinge region, thereby converting the F(ab)′ 2  dimer into a Fab′ monomer. The Fab′ monomer is essentially Fab with part of the hinge region (see, e.g.,  Fundamental Immunology  (Paul, editor, 7th edition, 2012)). While various antibody fragments are defined in terms of the digestion of an intact antibody, such fragments may be synthesized de novo either chemically or by using recombinant DNA methodology. Thus, the term antibody, as used herein, also includes antibody fragments either produced by the modification of whole antibodies, or those synthesized de novo using recombinant DNA methodologies (e.g., single chain Fv), or those identified using phage display libraries (see, e.g., McCafferty et al.,  Nature,  348: 552-554 (1990)). 
     The term “antibody” specifically encompasses monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments that exhibit the desired biological activity. 
     As used herein, the term “epitope” means any antigenic determinant or epitopic determinant of an antigen to which an antigen binding domain binds (i.e., at the 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, as well as specific charge characteristics. 
     As used herein, “HER2” refers to the protein human epidermal growth factor receptor 2 (SEQ ID NO: 1), or an antigen with least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more sequence identity to SEQ ID NO: 1. 
     Percent (%) identity of sequences can be calculated, for example, as 100×[(identical positions)/min(TG A , TG B )], where TG A  and TG B  are the sum of the number of residues and internal gap positions in peptide sequences A and B in the alignment that minimizes TG A  and TG B . See, e.g., Russell et al.,  J. Mol Biol.,  244: 332-350 (1994). 
     As used herein, the term “adjuvant” refers to a substance capable of eliciting an immune response in a subject exposed to the adjuvant. The phrase “adjuvant moiety” refers to an adjuvant that is covalently bonded to an antibody construct, e.g., through a linker, as described herein. The adjuvant moiety can elicit the immune response while bonded to the antibody construct or after cleavage (e.g., enzymatic cleavage) from the antibody construct following administration of an immunoconjugate to the subject. 
     As used herein, the terms “Toll-like receptor” and “TLR” refer to any member of a family of highly-conserved mammalian proteins which recognizes pathogen-associated molecular patterns and acts as key signaling elements in innate immunity. TLR polypeptides share a characteristic structure that includes an extracellular domain that has leucine-rich repeats, a transmembrane domain, and an intracellular domain that is involved in TLR signaling. 
     The terms “Toll-like receptor 7” and “TLR7” refer to nucleic acids or polypeptides sharing at least about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or more sequence identity to a publicly-available TLR7 sequence, e.g., GenBank accession number AAZ99026 for human TLR7 polypeptide, or GenBank accession number AAK62676 for murine TLR7 polypeptide. 
     The terms “Toll-like receptor 8” and “TLR8” refer to nucleic acids or polypeptides sharing at least about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or more sequence identity to a publicly-available TLR7 sequence, e.g., GenBank accession number AAZ95441 for human TLR8 polypeptide, or GenBank accession number AAK62677 for murine TLR8 polypeptide. 
     A “TLR agonist” is a substance that binds, directly or indirectly, to a TLR (e.g., TLR7 and/or TLR8) to induce TLR signaling. Any detectable difference in TLR signaling can indicate that an agonist stimulates or activates a TLR. Signaling differences can be manifested, for example, as changes in the expression of target genes, in the phosphorylation of signal transduction components, in the intracellular localization of downstream elements such as nuclear factor-κB (NF-κB), in the association of certain components (such as IL-1 receptor associated kinase (IRAK)) with other proteins or intracellular structures, or in the biochemical activity of components such as kinases (such as mitogen-activated protein kinase (MAPK)). 
     As used herein, “Ab” refers to an antibody construct that has an antigen-binding domain that binds HER2 (e.g., trastuzumab (also known as HERCEPTIN™), pertuzumab, biosimilars thereof, or biobetters thereof). 
     As used herein, the term “biosimilar” refers to an approved antibody construct that has active properties similar to the antibody construct previously approved (e.g., trastuzumab or pertuzumab). 
     As used herein, the term “biobetter” refers to an approved antibody construct that is an improvement of a previously approved antibody construct (e.g., trastuzumab or pertuzumab). The biobetter can have one or more modifications (e.g., an altered glycan profile, or a unique epitope) over the previously approved antibody construct. 
     As used herein, the term “amino acid” refers to any monomeric 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 unnatural (non-naturally occurring) amino acids and their stereoisomers. “Stereoisomers” of a given amino acid refer to isomers having the same molecular formula and intramolecular bonds but different three-dimensional arrangements of bonds and atoms (e.g., an L-amino acid and the corresponding D-amino acid). The amino acids can be glycosylated (e.g., N-linked glycans, O-linked glycans, phosphoglycans, C-linked glycans, or glypiation) or deglycosylated. 
     Naturally-occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Naturally-occurring α-amino acids include, without limitation, 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), tryptophan (Trp), tyrosine (Tyr), and combinations thereof. Stereoisomers of naturally-occurring α-amino acids include, without limitation, D-alanine (D-Ala), D-cysteine (D-Cys), D-aspartic acid (D-Asp), 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. 
     Unnatural (non-naturally occurring) amino acids include, without limitation, amino acid analogs, amino acid mimetics, synthetic amino acids, N-substituted glycines, and N-methyl amino acids in either the L- or D-configuration that function in a manner similar to the naturally-occurring amino acids. For example, “amino acid analogs” can be unnatural amino acids that have the same basic chemical structure as naturally-occurring amino acids (i.e., a carbon that is bonded to a hydrogen, a carboxyl group, an amino group) but have modified side-chain groups or modified peptide backbones, e.g., homoserine, norleucine, methionine sulfoxide, and methionine methyl sulfonium. “Amino acid mimetics” refer to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally-occurring amino acid. 
     Amino acids may be referred to herein by either the commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. 
     As used herein, the term “linker” refers to a functional group that covalently bonds two or more moieties in a compound or material. For example, the linking moiety can serve to covalently bond an adjuvant moiety to an antibody construct in an immunoconjugate. 
     As used herein, the term “linking moiety” refers to a functional group that covalently bonds two or more moieties in a compound or material. For example, the linking moiety can serve to covalently bond an adjuvant moiety to an antibody in an immunoconjugate. Useful bonds for connecting linking moieties to proteins and other materials include, but are not limited to, amides, amines, esters, carbamates, ureas, thioethers, thiocarbamates, thiocarbonates, and thioureas. 
     As used herein, the term “divalent” refers to a chemical moiety that contains two points of attachment for linking two functional groups; polyvalent linking moieties can have additional points of attachment for linking further functional groups. For example, divalent linking moieties include divalent polymer moieties such as divalent poly(ethylene glycol), divalent cycloalkyl, divalent heterocycloalkyl, divalent aryl, and divalent heteroaryl group. A “divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group” refers to a cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group having two points of attachment for covalently linking two moieties in a molecule or material. Cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups can be substituted or unsubstituted. Cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups can be substituted with one or more groups selected from halo, hydroxy, amino, alkylamino, amido, acyl, nitro, cyano, and alkoxy. 
     As used herein, when the term “optionally present” is used to refer to a chemical structure (e.g., “A”, “U”, “V”, “X 1 ”, “X 2 ”, and “X 3 ”), if that chemical structure is not present, the bond originally made to the chemical structure is made directly to the adjacent atom. 
     As used herein, the wavy line (“ ”) represents a point of attachment of the specified chemical moiety. If the specified chemical moiety has two wavy lines (“ ”) present, it will be understood that the chemical moiety can be used bilaterally, i.e., as read from left to right or from right to left. In some embodiments, a specified moiety having two wavy lines (“ ”) present is considered to be used as read from left to right. 
     As used herein, the term “linker” refers to a functional group that covalently bonds two or more moieties in a compound or material. For example, the linker can serve to covalently bond an adjuvant moiety to an antibody construct in an immunoconjugate. 
     As used herein, the term “alkyl” refers to a straight or branched, saturated, aliphatic radical having the number of carbon atoms indicated. Alkyl can include any number of carbons, such as C 1 -C 2 , C 1 -C 3 , C 1 -C 4 , C 1 -C 5 , C 1 -C 6 , C 1 -C 7 , C 1 -C 8 , C 1 -C 9 , C 1 -C 10 , C 2 -C 3 , C 2 -C 4 , C 2 -C 5 , C 2 -C 6 , C 3 -C 4 , C 3 -C 5 , C 3 -C 6 , C 4 -C 5 , C 4 -C 6  and C 5 -C 6 . For example, C 1 -C 4  alkyl includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl. Alkyl can also refer to alkyl groups having up to 30 carbons atoms, such as, but not limited to heptyl, octyl, nonyl, decyl, etc. Alkyl groups can be substituted or unsubstituted. “Substituted alkyl” groups can be substituted with one or more groups selected from halo, hydroxy, amino, oxo (═O), alkylamino, amido, acyl, nitro, cyano, and alkoxy. The term “alkylene” refers to a divalent alkyl radical. 
     As used herein, the term “heteroalkyl” refers to an alkyl group as described herein, wherein one or more carbon atoms are optionally and independently replaced with heteroatom selected from N, O, and S. The term “heteroalkylene” refers to a divalent heteroalkyl radical. 
     As used herein, the term “cycloalkyl” refers to a saturated or partially unsaturated, monocyclic, fused bicyclic, or bridged polycyclic ring assembly containing from 3 to 12 ring atoms, or the number of atoms indicated. Carbocycles can include any number of carbons, such as C 3 -C 6 , C 4 -C 6 , C 5 -C 6 , C 3 -C 8 , C 4 -C 8 , C 5 -C 8 , C 6 -C 8 , C 3 -C 9 , C 3 -C 10 , C 3 -C 11 , and C 3 -C 12 . 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 in the ring. Representative carbocyclic groups that are partially unsaturated include, but are not limited to, cyclobutene, cyclopentene, cyclohexene, cyclohexadiene (1,3- and 1,4-isomers), cycloheptene, cycloheptadiene, cyclooctene, cyclooctadiene (1,3-, 1,4- and 1,5-isomers), norbornene, and norbornadiene. 
     As used herein, the term “aryl” refers to an aromatic ring system having any suitable number of ring atoms and any suitable number of rings. Aryl groups can include any suitable number of ring atoms, such as, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 ring atoms, as well as from 6 to 10, 6 to 12, or 6 to 14 ring members. Aryl groups can be monocyclic, fused to form bicyclic or tricyclic groups, or linked by a bond to form a biaryl group. Representative aryl groups include phenyl, naphthyl and biphenyl. Other aryl groups include benzyl, having a methylene linking group. Some aryl groups have from 6 to 12 ring members, such as phenyl, naphthyl or biphenyl. Other aryl groups have from 6 to 10 ring members, such as phenyl or naphthyl. 
     As used herein, the terms “heterocycloalkyl” and “heteroaryl” refer to a “cycloalkyl” or “aryl” group as described herein, wherein one or more carbon atoms are optionally and independently replaced with heteroatom selected from N, O, and S. “Heteroaryl,” by itself or as part of another substituent, refers to a monocyclic or fused bicyclic or tricyclic aromatic ring assembly containing 5 to 16 ring atoms, where from 1 to 5 of the ring atoms are a heteroatom such as N, O or S. Additional heteroatoms can also be useful, including, but not limited to, B, Al, Si and P. The heteroatoms can be oxidized to form moieties such as, but not limited to, —S(O)— and —S(O) 2 —. Heteroaryl groups can include any number of ring atoms, such as 3 to 6, 4 to 6, 5 to 6, 3 to 8, 4 to 8, 5 to 8, 6 to 8, 3 to 9, 3 to 10, 3 to 11, or 3 to 12 ring members. Any suitable number of heteroatoms can be included in the heteroaryl groups, such as 1, 2, 3, 4, or 5, or 1 to 2, 1 to 3, 1 to 4, 1 to 5, 2 to 3, 2 to 4, 2 to 5, 3 to 4, or 3 to 5. The heteroaryl group can include groups such as pyrrole, pyridine, imidazole, pyrazole, triazole, tetrazole, pyrazine, pyrimidine, pyridazine, triazine (1,2,3-, 1,2,4- and 1,3,5-isomers), thiophene, furan, thiazole, isothiazole, oxazole, and isoxazole. The heteroaryl groups can also be fused to aromatic ring systems, such as a phenyl ring, to form members including, but not limited to, benzopyrroles such as indole and isoindole, benzopyridines such as quinoline and isoquinoline, benzopyrazine (quinoxaline), benzopyrimidine (quinazoline), benzopyridazines such as phthalazine and cinnoline, benzothiophene, and benzofuran. Other heteroaryl groups include heteroaryl rings linked by a bond, such as bipyridine. Heteroaryl groups can be substituted or unsubstituted. “Substituted heteroaryl” groups can be substituted with one or more groups selected from halo, hydroxy, amino, oxo (═O), alkylamino, amido, acyl, nitro, cyano, and alkoxy. 
     Heteroaryl groups can be linked via any position on the ring. For example, pyrrole includes 1-, 2- and 3-pyrrole, pyridine includes 2-, 3- and 4-pyridine, imidazole includes 1-, 2-, 4- and 5-imidazole, pyrazole includes 1-, 3-, 4- and 5-pyrazole, triazole includes 1-, 4- and 5-triazole, tetrazole includes 1- and 5-tetrazole, pyrimidine includes 2-, 4-, 5- and 6-pyrimidine, pyridazine includes 3- and 4-pyridazine, 1,2,3-triazine includes 4- and 5-triazine, 1,2,4-triazine includes 3-, 5- and 6-triazine, 1,3,5-triazine includes 2-triazine, thiophene includes 2- and 3-thiophene, furan includes 2- and 3-furan, thiazole includes 2-, 4- and 5-thiazole, isothiazole includes 3-, 4- and 5-isothiazole, oxazole includes 2-, 4- and 5-oxazole, isoxazole includes 3-, 4- and 5-isoxazole, indole includes 1-, 2- and 3-indole, isoindole includes 1- and 2-isoindole, quinoline includes 2-, 3- and 4-quinoline, isoquinoline includes 1-, 3- and 4-isoquinoline, quinazoline includes 2- and 4-quinoazoline, cinnoline includes 3- and 4-cinnoline, benzothiophene includes 2- and 3-benzothiophene, and benzofuran includes 2- and 3-benzofuran. 
     “Heterocycloalkyl,” by itself or as part of another substituent, refers to a saturated ring system having from 3 to 12 ring members and from 1 to 4 heteroatoms of N, O and S. Additional heteroatoms can also be useful, including, but not limited to, B, Al, Si and P. The heteroatoms can be oxidized to form moieties such as, but not limited to, —S(O)— and —S(O) 2 —. Heterocycloalkyl groups can include any number of ring atoms, such as, 3 to 6, 4 to 6, 5 to 6, 3 to 8, 4 to 8, 5 to 8, 6 to 8, 3 to 9, 3 to 10, 3 to 11, or 3 to 12 ring members. Any suitable number of heteroatoms can be included in the heterocycloalkyl groups, such as 1, 2, 3, or 4, or 1 to 2, 1 to 3, 1 to 4, 2 to 3, 2 to 4, or 3 to 4. The heterocycloalkyl group can include groups such as aziridine, azetidine, pyrrolidine, piperidine, azepane, azocane, quinuclidine, pyrazolidine, imidazolidine, piperazine (1,2-, 1,3- and 1,4-isomers), oxirane, oxetane, tetrahydrofuran, oxane (tetrahydropyran), oxepane, thiirane, thietane, thiolane (tetrahydrothiophene), thiane (tetrahydrothiopyran), oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, dioxolane, dithiolane, morpholine, thiomorpholine, dioxane, or dithiane. The heterocycloalkyl groups can also be fused to aromatic or non-aromatic ring systems to form members including, but not limited to, indoline. Heterocycloalkyl groups can be unsubstituted or substituted. 
     Heterocycloalkyl groups can be linked via any position on the ring. For example, aziridine can be 1- or 2-aziridine, azetidine can be 1- or 2-azetidine, pyrrolidine can be 1-, 2- or 3-pyrrolidine, piperidine can be 1-, 2-, 3- or 4-piperidine, pyrazolidine can be 1-, 2-, 3-, or 4-pyrazolidine, imidazolidine can be 1-, 2-, 3- or 4-imidazolidine, piperazine can be 1-, 2-, 3- or 4-piperazine, tetrahydrofuran can be 1- or 2-tetrahydrofuran, oxazolidine can be 2-, 3-, 4- or 5-oxazolidine, isoxazolidine can be 2-, 3-, 4- or 5-isoxazolidine, thiazolidine can be 2-, 3-, 4- or 5-thiazolidine, isothiazolidine can be 2-, 3-, 4- or 5-isothiazolidine, and morpholine can be 2-, 3- or 4-morpholine. 
     As used herein, the terms “halo” and “halogen,” by themselves or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom. 
     As used herein, the term “carbonyl,” by itself or as part of another substituent, refers to C(O) or —C(O)—, i.e., a carbon atom double-bonded to oxygen and bound to two other groups in the moiety having the carbonyl. 
     As used herein, the term “amino” refers to a moiety —NR3, wherein each R group is H or alkyl. An amino moiety can be ionized to form the corresponding ammonium cation. 
     As used herein, the phrase “quaternary ammonium salt” refers to a tertiary amine that has been quaternized with an alkyl substituent (e.g., a C 1 -C 4  alkyl such as methyl, ethyl, propyl, or butyl). 
     As used herein, the term “hydroxyl” refers to the moiety —OH. 
     As used herein, the term “cyano” refers to a carbon atom triple-bonded to a nitrogen atom (i.e., the moiety —C≡N). 
     As used herein, the terms “treat,” “treatment,” and “treating” refer to any indicia of success in the treatment or amelioration of an injury, pathology, condition (e.g., cancer), or symptom (e.g., cognitive impairment), including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the symptom, injury, pathology, or condition more tolerable to the patient; reduction in the rate of symptom progression; decreasing the frequency or duration of the symptom or condition; or, in some situations, preventing the onset of the symptom. The treatment or amelioration of symptoms can be based on any objective or subjective parameter, including, for example, the result of a physical examination. 
     The terms “cancer,” “neoplasm,” and “tumor” are used herein to refer to cells which exhibit autonomous, unregulated growth, such that the cells exhibit an aberrant growth phenotype characterized by a significant loss of control over cell proliferation. Cells of interest for detection, analysis, and/or treatment in the context of the invention include cancer cells (e.g., cancer cells from an individual with cancer), malignant cancer cells, pre-metastatic cancer cells, metastatic cancer cells, and non-metastatic cancer cells. Cancers of virtually every tissue are known. The phrase “cancer burden” refers to the quantum of cancer cells or cancer volume in a subject. Reducing cancer burden accordingly refers to reducing the number of cancer cells or the cancer cell volume in a subject. The term “cancer cell” as used herein refers to any cell that is a cancer cell (e.g., from any of the cancers for which an individual can be treated, e.g., isolated from an individual having cancer) or is derived from a cancer cell, e.g., clone of a cancer cell. For example, a cancer cell can be from an established cancer cell line, can be a primary cell isolated from an individual with cancer, can be a progeny cell from a primary cell isolated from an individual with cancer, and the like. In some embodiments, the term can also refer to a portion of a cancer cell, such as a sub-cellular portion, a cell membrane portion, or a cell lysate of a cancer cell. Many types of cancers are known to those of skill in the art, including solid tumors such as carcinomas, sarcomas, glioblastomas, melanomas, lymphomas, and myelomas, and circulating cancers such as leukemias. 
     As used herein, the term “cancer” includes any form of cancer, including but not limited to, solid tumor cancers (e.g., lung, prostate, breast, gastric, bladder, colon, ovarian, pancreas, kidney, liver, glioblastoma, medulloblastoma, leiomyosarcoma, head &amp; neck squamous cell carcinomas, melanomas, and neuroendocrine) and liquid cancers (e.g., hematological cancers); carcinomas; soft tissue tumors; sarcomas; teratomas; melanomas; leukemias; lymphomas; and brain cancers, including minimal residual disease, and including both primary and metastatic tumors. Any HER2 expressing cancer is a suitable cancer to be treated by the subject methods and compositions. As used herein “HER2 expression” refers to a cell that has a HER2 receptor on the cell&#39;s surface. For example, a cell may have from about 20,000 to about 50,000 HER2 receptors on the cell&#39;s surface. As used herein “HER2 overexpression” refers to a cell that has more than about 50,000 HER2 receptors. For example, a cell 2, 5, 10, 100, 1,000, 10,000, 100,000, or 1,000,000 times the number of HER2 receptors as compared to corresponding non-cancer cell (e.g., about 1 or 2 million HER2 receptors). It is estimated that HER2 is overexpressed in about 25% to about 30% of breast cancers. 
     Carcinomas are malignancies that originate in the epithelial tissues. Epithelial cells cover the external surface of the body, line the internal cavities, and form the lining of glandular tissues. Examples of carcinomas include, but are not limited to, adenocarcinoma (cancer that begins in glandular (secretory) cells such as cancers of the breast, pancreas, lung, prostate, stomach, gastroesophageal junction, and colon) adrenocortical carcinoma; hepatocellular carcinoma; renal cell carcinoma; ovarian carcinoma; carcinoma in situ; ductal carcinoma; carcinoma of the breast; basal cell carcinoma; squamous cell carcinoma; transitional cell carcinoma; colon carcinoma; nasopharyngeal carcinoma; multilocular cystic renal cell carcinoma; oat cell carcinoma; large cell lung carcinoma; small cell lung carcinoma; non-small cell lung carcinoma; and the like. Carcinomas may be found in prostrate, pancreas, colon, brain (usually as secondary metastases), lung, breast, and skin. 
     Soft tissue tumors are a highly diverse group of rare tumors that are derived from connective tissue. Examples of soft tissue tumors include, but are not limited to, alveolar soft part sarcoma; angiomatoid fibrous histiocytoma; chondromyoxid fibroma; skeletal chondrosarcoma; extraskeletal myxoid chondrosarcoma; clear cell sarcoma; desmoplastic small round-cell tumor; dermatofibrosarcoma protuberans; endometrial stromal tumor; Ewing&#39;s sarcoma; fibromatosis (Desmoid); fibrosarcoma, infantile; gastrointestinal stromal tumor; bone giant cell tumor; tenosynovial giant cell tumor; inflammatory myofibroblastic tumor; uterine leiomyoma; leiomyosarcoma; lipoblastoma; typical lipoma; spindle cell or pleomorphic lipoma; atypical lipoma; chondroid lipoma; 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; embryonal rhabdomyosarcoma; benign or malignant schwannoma; synovial sarcoma; Evan&#39;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); fibrous dysplasia; myxofibrosarcoma; fibrosarcoma; synovial sarcoma; malignant peripheral nerve sheath tumor; neurofibroma; pleomorphic adenoma of soft tissue; and neoplasias derived from fibroblasts, myofibroblasts, histiocytes, vascular cells/endothelial cells, and nerve sheath cells. 
     A sarcoma is a rare type of cancer that arises in cells of mesenchymal origin, e.g., in bone or in the soft tissues of the body, including cartilage, fat, muscle, blood vessels, fibrous tissue, or other connective or supportive tissue. Different types of sarcoma are based on where the cancer forms. For example, osteosarcoma forms in bone, liposarcoma forms in fat, and rhabdomyosarcoma forms in muscle. Examples of sarcomas include, but are not limited to, askin&#39;s tumor; sarcoma botryoides; chondrosarcoma; ewing&#39;s sarcoma; malignant hemangioendothelioma; malignant schwannoma; osteosarcoma; and soft tissue sarcomas (e.g., alveolar soft part sarcoma; angiosarcoma; cystosarcoma phyllodesdermatofibrosarcoma protuberans (DFSP); desmoid tumor; desmoplastic small round cell tumor; epithelioid sarcoma; extraskeletal chondrosarcoma; extraskeletal osteosarcoma; fibrosarcoma; gastrointestinal stromal tumor (GIST); hemangiopericytoma; hemangiosarcoma (more commonly referred to as “angiosarcoma”); kaposi&#39;s sarcoma; leiomyosarcoma; liposarcoma; lymphangiosarcoma; malignant peripheral nerve sheath tumor (MPNST); neurofibrosarcoma; synovial sarcoma; and undifferentiated pleomorphic sarcoma). 
     A teratoma is a type of germ cell tumor that may contain several different types of tissue (e.g., can include tissues derived from any and/or all of the three germ layers: endoderm, mesoderm, and ectoderm), including, for example, hair, muscle, and bone. Teratomas occur most often in the ovaries in women, the testicles in men, and the tailbone in children. 
     Melanoma is a form of cancer that begins in melanocytes (cells that make the pigment melanin). Melanoma may begin in a mole (skin melanoma), but can also begin in other pigmented tissues, such as in the eye or in the intestines. 
     Leukemias are cancers that start in blood-forming tissue, such as the bone marrow, and cause large numbers of abnormal blood cells to be produced and enter the bloodstream. For example, leukemias can originate in bone marrow-derived cells that normally mature in the bloodstream. Leukemias are named for how quickly the disease develops and progresses (e.g., acute versus chronic) and for the type of white blood cell that is affected (e.g., myeloid versus lymphoid). Myeloid leukemias are also called myelogenous or myeloblastic leukemias. Lymphoid leukemias are also called lymphoblastic or lymphocytic leukemia. Lymphoid leukemia cells may collect in the 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). 
     Lymphomas are cancers that begin in cells of the immune system. For example, lymphomas can originate in bone marrow-derived cells that normally mature in the lymphatic system. There are two basic categories of lymphomas. One category of lymphoma is Hodgkin lymphoma (HL), which is marked by the presence of a type of cell called the Reed-Sternberg cell. There are currently 6 recognized types of HL. Examples of Hodgkin lymphomas include nodular sclerosis classical Hodgkin lymphoma (CHL), mixed cellularity CHL, lymphocyte-depletion CHL, lymphocyte-rich CHL, and nodular lymphocyte predominant HL. 
     The other category of lymphoma is non-Hodgkin lymphomas (NHL), which includes a large, diverse group of cancers of immune system cells. Non-Hodgkin lymphomas can be further divided into cancers that have an indolent (slow-growing) course and those that have an aggressive (fast-growing) course. There are currently 61 recognized types of NHL. Examples of non-Hodgkin lymphomas include, but are not limited to, AIDS-related Lymphomas, anaplastic large-cell lymphoma, angioimmunoblastic lymphoma, blastic NK-cell lymphoma, Burkitt&#39;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, enteropathy-type T-Cell lymphoma, follicular lymphoma, hepatosplenic gamma-delta T-Cell lymphomas, T-Cell leukemias, lymphoblastic lymphoma, mantle cell lymphoma, marginal zone lymphoma, nasal T-Cell lymphoma, pediatric lymphoma, peripheral T-Cell lymphomas, primary central nervous system lymphoma, transformed lymphomas, treatment-related T-Cell lymphomas, and Waldenstrom&#39;s macroglobulinemia. 
     Brain cancers include any cancer of the brain tissues. Examples of brain cancers include, but are not limited to, gliomas (e.g., glioblastomas, astrocytomas, oligodendrogliomas, ependymomas, and the like), meningiomas, pituitary adenomas, and vestibular schwannomas, primitive neuroectodermal tumors (medulloblastomas). 
     The “pathology” of cancer includes all phenomena that compromise the well-being of the patient. This includes, without limitation, abnormal or uncontrollable cell growth, metastasis, interference with the normal functioning of neighboring cells, release of cytokines or other secretory products at abnormal levels, suppression or aggravation of inflammatory or immunological response, neoplasia, premalignancy, malignancy, and invasion of surrounding or distant tissues or organs, such as lymph nodes. 
     As used herein, the phrases “cancer recurrence” and “tumor recurrence,” and grammatical variants thereof, refer to further growth of neoplastic or cancerous cells after diagnosis of cancer. Particularly, recurrence may occur when further cancerous cell growth occurs in the cancerous tissue. “Tumor spread,” similarly, occurs when the cells of a tumor disseminate into local or distant tissues and organs, therefore, tumor spread encompasses tumor metastasis. “Tumor invasion” occurs when the tumor growth spread out locally to compromise the function of involved tissues by compression, destruction, or prevention of normal organ function. 
     As used herein, the term “metastasis” refers to the growth of a cancerous tumor in an organ or body part, which is not directly connected to the organ of the original cancerous tumor. Metastasis will be understood to include micrometastasis, which is the presence of an undetectable amount 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 of a process, such as the departure of cancer cells from an original tumor site, and migration and/or invasion of cancer cells to other parts of the body. 
     As used herein the phrases “effective amount” and “therapeutically effective amount” refer to a dose of a substance such as an immunoconjugate that produces therapeutic effects for which it is administered. The exact dose will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, 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  &amp;  Gilman&#39;s The Pharmacological Basis of Therapeutics,  11 th  Edition (McGraw-Hill, 2006); and  Remington: The Science and Practice of Pharmacy,  22 nd  Edition, (Pharmaceutical Press, London, 2012)). 
     As used herein, the terms “recipient,” “individual,” “subject,” “host,” and “patient” are used interchangeably and refer to any mammalian subject for whom diagnosis, treatment, or therapy is desired (e.g., humans). “Mammal” for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, horses, cats, cows, sheep, goats, pigs, camels, etc. In certain embodiments, the mammal is human. 
     The phrase “synergistic adjuvant” or “synergistic combination” in the context of this invention includes the combination of two immune modulators such as a receptor agonist, cytokine, and adjuvant polypeptide, that in combination elicit a synergistic effect on immunity relative to either administered alone. Particularly, the immunoconjugates disclosed herein comprise synergistic combinations of the claimed adjuvant and antibody construct. These synergistic combinations upon administration elicit a greater effect on immunity, e.g., relative to when the antibody construct or adjuvant is administered in the absence of the other moiety. Further, a decreased amount of the immunoconjugate may be administered (as measured by the total number of antibody constructs or the total number of adjuvants administered as part of the immunoconjugate) compared to when either the antibody construct or adjuvant is administered alone. 
     As used herein, the term “administering” refers to parenteral, intravenous, intraperitoneal, intramuscular, intratumoral, intralesional, intranasal, or subcutaneous administration, oral administration, administration as a suppository, topical contact, intrathecal administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to the subject. 
     The terms “about” and “around,” as used herein to modify a numerical value, indicate a close range surrounding the numerical value. Thus, if “X” is the value, “about X” or “around X” indicates a value of from 0.9X to 1.1X, e.g., from 0.95X to 1.05X or from 0.99X to 1.01X. A reference to “about X” or “around X” specifically indicates at least the values X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, and 1.05X. Accordingly, “about X” and “around X” are intended to teach and provide written description support for a claim limitation of, e.g., “0.98X.” 
     Antibody Adjuvant Conjugates 
     The invention provides an immunoconjugate of formula (I): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 1  is CH or N, 
     J 2  is CHQ, NQ, O, or S, 
     each Q independently is Y or Z, wherein exactly one Q is Y, 
     Y is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. “Ab” can be any suitable antibody construct that has an antigen binding domain that binds HER2, such as, for example, trastuzumab and pertuzumab. In certain embodiments, “Ab” is trastuzumab (also known as HERCEPTIN™), a biosimilar thereof, or a biobetter thereof. For example, “Ab” can be MYL-14010, ABP 980, BCD-022, CT-P6, EG12014, HD201, ONS-1050, PF-05280014, Ontruzant, Saiputing, Herzuma, or HLX02. In preferred embodiments, “Ab” is trastuzumab (also known as HERCEPTIN™). 
     In certain embodiments, the immunoconjugate is of formula (Ia): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 2  is CHZ, NZ, O, or S, 
     Y is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula. 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ia 1 ), (Ia 2 ), (Ia 3 ), (Ia 4 ), (Ia 5 ), or (Ia 6 ): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Iaa), (Iab), (Iac), (Iad), (Iae), or (Iaf): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 2  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 4  is CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ib): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula. 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ib 1 ), (Ib 2 ), (Ib 3 ), or (Ib 4 ): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Iba), (Ibb), (Ibc), or (Ibd): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 2  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ic): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 1  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  is an integer from 1 to 25, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     X 1 , X 2 , and X 3  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 2  is an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Id): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 1  is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  is an integer from 1 to 25, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     X 1 , X 2 , and X 3  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 2  is an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ie): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 2  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  is hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (If): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 2  is of formula: 
     
       
         
         
             
             
         
       
     
     or 
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  is hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of 
     m 1 , m 2 , and m 3  is a non-zero integer, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ig): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 3  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  is hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ih): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 3  is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  is hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ii): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 4  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  is an integer from 1 to 25, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     X 1 , X 2 , and X 3  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 2  is an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ij): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 4  is of formula: 
     
       
         
         
             
             
         
       
     
     or 
     each Z independently is hydrogen or of formula. 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  is an integer from 1 to 25, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     X 1 , X 2 , and X 3  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 2  is an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Ik): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 5  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  and m 2  independently are an integer from 0 to 25, except that at least one of m 1  and m 2  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (Im): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 5  is of formula: 
     
       
         
         
             
             
         
       
     
     or 
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  and m 2  independently are an integer from 0 to 25, except that at least one of m 1  and m 2  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     The invention further provides an immunoconjugate of formula (II): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     each Q independently is Y or Z, wherein exactly one Q is Y, 
     Y is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula. 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. “Ab” can be any suitable antibody construct that has an antigen binding domain that binds HER2, such as, for example, trastuzumab and pertuzumab. In certain embodiments, “Ab” is trastuzumab (also known as HERCEPTIN™), a biosimilar thereof, or a biobetter thereof. For example, “Ab” can be MYL-14010, ABP 980, BCD-022, CT-P6, EG12014, HD201, ONS-1050, PF-05280014, Ontruzant, Saiputing, Herzuma, or HLX02. In preferred embodiments, “Ab” is trastuzumab (also known as HERCEPTIN™) 
     In certain embodiments, the immunoconjugate is of formula (IIa): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIa 1 ), (IIa 2 ), (IIa 3 ), (IIa 4 ), (IIa 5 ), or (IIa 6 ): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIaa), (IIab), (IIac), (IIad), (IIae), or (IIaf): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 2  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 4  is CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIb): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIb 1 ), (IIb 2 ), (IIb 3 ), or (IIb 4 ): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIba), (IIbb), (IIbc), or (IIbd): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 2  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIc): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 1  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  is an integer from 1 to 25, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     X 1 , X 2 , and X 3  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 2  is an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IId): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 1  is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  is an integer from 1 to 25, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     X 1 , X 2 , and X 3  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 2  is an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIe): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 2  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  is hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (If): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 2  is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  is hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIg): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 3  is of formula: 
     
       
         
         
             
             
         
       
     
     R 6  is hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIh): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 3  is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  is hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIi): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 4  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  is an integer from 1 to 25, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     X 1 , X 2 , and X 3  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 2  is an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIj): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 4  is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula. 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  is an integer from 1 to 25, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     X 1 , X 2 , and X 3  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 2  is an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIk): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 5  is of formula: 
     
       
         
         
             
             
         
       
     
     Z is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     J P  and J 4  independently are CH or N, 
     m 1  and m 2  independently are an integer from 0 to 25, except that at least one of m 1  and m 2  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments, the immunoconjugate is of formula (IIm): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     Y 5  is of formula: 
     
       
         
         
             
             
         
       
     
     or 
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1  and m 2  independently are an integer from 0 to 25, except that at least one of m 1  and m 2  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     p is an integer from 1 to 4, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 4  is CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and 
     each wavy line (“ ”) represents a point of attachment. 
     In certain embodiments of the invention, one or more aromatic hydrogen atoms in formulas (I) and (II) can be substituted with a halogen atom (e.g., fluorine, chlorine, bromine, iodine, or combinations thereof). 
     For variable Y described herein as formulas: 
     
       
         
         
             
             
         
       
     
     it will be understood that the structures: 
     
       
         
         
             
             
         
       
     
     are present within the brackets such that the variable r also applies to said structures. 
     Generally, the immunoconjugates of the invention comprise about 1 to about 10 adjuvants, each adjuvant linked to the antibody construct, as designated with subscript “r”. In an embodiment, r is 1, such that there is a single adjuvant linked to the antibody construct. In some embodiments, r is an integer from about 2 to about 10 (e.g., about 2 to about 9, about 3 to about 9, about 4 to about 9, about 5 to about 9, about 6 to about 9, about 3 to about 8, about 3 to about 7, about 3 to about 6, about 4 to about 8, about 4 to about 7, about 4 to about 6, about 5 to about 6, about 1 to about 6, about 1 to about 4, about 2 to about 4, or about 1 to about 3). Accordingly, the immunoconjugates can have (i.e., subscript “r” can be) 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 adjuvants linked to the antibody construct. In preferred embodiments, the immunoconjugates have (i.e., subscript “r” can be) 1, 2, 3, or 4 adjuvants linked to the antibody construct. The desirable adjuvant to antibody construct ratio (i.e., the value of the subscript “r”) can be determined by a skilled artisan depending on the desired effect of the treatment. 
     In some embodiments, X 1 , X 2 , X 3 , and X 4  independently are one or more divalent groups selected from benzene, naphthalene, pyrrole, indole, isoindole, indolizine, furan, benzofuran, benzothiophene, thiophene, pyridine, acridine, naphthyridine, quinolone, isoquinoline, isoxazole, oxazole, benzoxazole, isothiazole, thiazole, benzthiazole, imidazole, thiadiazole, tetrazole, triazole, oxadiazole, benzimidazole, purine, pyrazole, pyrazine, pteridine, quinoxaline, phthalazine, quinazoline, triazine, phenazine, cinnoline, pyrimidine, pyridazine, cyclohexane, decahydronaphthalene, pyrrolidine, octahydroindole, octahydroisoindole, tetrahydrofuran, octahydrobenzofuran, octahydrobenzothiophene, tetrahydrothiophene, piperidine, tetradecahydroacridine, naphthyridine, decahydroquinoline, decahydroisoquinoline, isoxazolidine, oxazolidine, octahydrobenzooxazole, isothiazolidine, thiazolidine, octahydrobenzothiazole, imidazolidine, 1,2,3-thiadiazolidine, tetrazolidine, 1,2,3-triazolidine, 1,2,3-oxadiazolidine, octahydrobenzoimidazole, octahydropurine, pyrazolidine, piperazine, dechydropteridine, decahydroquinoxaline, dechydrophthalazine, dechydroquinazoline, 1,3,5-triazinane, tetradecahydrophenazine, decahydrocinnoline, hexhydropyrimidine, or hexahydropyridazine. In some embodiments, the one or more divalent groups of X 1 , X 2 , and X 3  are fused. In some embodiments, the one or more divalent groups of X 1 , X 2 , and X 3  are linked through a bond or —CO—. In certain embodiments, X 1 , X 2 , and X 3  can be substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof. 
     In certain embodiments, X 1 , X 2 , X 3 , and X 4  independently are of formula: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     wherein any of the above-referenced structures can be used bilaterally. 
     Variables Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof. Ar 1  and Ar 2  can be any suitable aryl or heteroaryl group described herein. In some embodiments, Ar 1  and Ar 2  independently are a monovalent aryl or heteroaryl group described by the divalent groups of, X 1 , X 2 , X 3 , and X 4 , optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof. 
     Variables m 1 , m 2 , and m 3  independently are an integer from 0 to 25. Typically, at least one of m 1 , m 2 , and m 3  is a non-zero integer such that at least one of m 1 , m 2 , and m 3  is an integer from 1 to 25. In certain embodiments, at least one of m 1 , m 2 , and m 3  is an integer from about 2 to about 25 (e.g., about 2 to about 16, about 6 to about 25, about 6 to about 16, about 8 to about 25, about 8 to about 16, about 6 to about 12, or about 8 to about 12). Accordingly, in some embodiments, the immunoconjugates of the invention comprise about 2 to about 25 (e.g., about 2 to about 16, about 6 to about 25, about 6 to about 16, about 8 to about 25, about 8 to about 16, about 6 to about 12, or about 8 to about 12) ethylene glycol units, as designated with subscripts “m 1 ”, “m 2 ” and “m 3 ”. Accordingly, the immunoconjugates of the invention can comprise at least 2 ethylene glycol groups (e.g., at least 3 ethylene glycol groups, at least 4 ethylene glycol groups, at least 5 ethylene glycol groups, at least 6 ethylene glycol groups, at least 7 ethylene glycol groups, at least 8 ethylene glycol groups, at least 9 ethylene glycol groups, or at least 10 ethylene glycol groups). Accordingly, the immunoconjugate can comprise from about 2 to about 25 ethylene glycol units, for example, from about 6 to about 25 ethylene glycol units, from about 6 to about 16 ethylene glycol units, from about 8 to about 25 ethylene glycol units, from about 8 to about 16 ethylene glycol units, from about 8 to about 12 ethylene glycol units, or from about 8 to about 12 ethylene glycol units. In certain embodiments, the immunoconjugate comprises a di(ethylene glycol) group, a tri(ethylene glycol) group, a tetra(ethylene glycol) group, 5 ethylene glycol groups, 6 ethylene glycol groups, 7 ethylene glycol groups, 8 ethylene glycol groups, 9 ethylene glycol groups, 10 ethylene glycol groups, 11 ethylene glycol groups, 12 ethylene glycol groups, 13 ethylene glycol groups, 14 ethylene glycol groups, 15 ethylene glycol groups, 16 ethylene glycol groups, 24 ethylene glycol groups, or 25 ethylene glycol groups. Variable p is an integer from 1 to 4 (e.g., 1, 2, 3, or 4). In certain embodiments, p is 1 such that the aryl ring has one Z substituent that is not hydrogen at one of the four available carbons. 
     Variables t 1  and t 2  independently are an integer from 1 to 3. In some embodiments, when present, t 1  and t 2  are 1 and 2, respectively, or t 1  and t 2  are 2 and 2, respectively. In preferred embodiments, when present, t 1  and t 2  are 2 and 2, respectively. 
     The linking moiety (L M ) represents the remnants of a chemical species used to conjugate the adjuvant to the antibody. L M  can be any suitable remnants of any conjugation techniques known in the art. One of skill in the art will appreciate that the adjuvant moieties in the conjugates can be covalently bonded to the antibodies using various chemistries for protein modification, and that the linking moieties described above result from the reaction of protein functional groups (i.e., amino acid side chains), with reagents having reactive linker groups. A wide variety of such reagents are known in the art. Examples of such reagents include, but are not limited to, N-hydroxysuccinimidyl (NHS) esters and N-hydroxysulfosuccinimidyl (sulfo-NHS) esters (amine reactive); carbodiimides (amine and carboxyl reactive); hydroxymethyl phosphines (amine reactive); maleimides (thiol reactive); halogenated acetamides such as N-iodoacetamides (thiol reactive); aryl azides (primary amine reactive); fluorinated aryl azides (reactive via carbon-hydrogen (C—H) insertion); pentafluorophenyl (PFP) esters (amine reactive); tetrafluorophenyl (TFP) esters (amine reactive); imidoesters (amine reactive); isocyanates (hydroxyl reactive); vinyl sulfones (thiol, amine, and hydroxyl reactive); pyridyl disulfides (thiol reactive); and benzophenone derivatives (reactive via C—H bond insertion). Further reagents include but are not limited to those described in Hermanson,  Bioconjugate Techniques  2nd Edition, Academic Press, 2008. 
     Linkers containing maleimide groups, vinyl sulfone groups, pyridyl disulfide groups, and halogenated acetamide groups are particularly useful for covalent bonding to thiol groups in an antibody. Thiol groups in an antibody are generally located in cysteine sidechains. Free thiol groups may be present in naturally-occurring, solvent-accessible cysteine residues in the antibody. Free thiols can also be present in engineered cysteine residues, as described below. In addition, thiol groups can be generated via full or partial reduction of disulfide linkages between cysteine sidechains in an antibody. Thiol groups can be also appended to lysine sidechains using known methods with reagents including, but not limited to, 2-iminothiolane (Traut&#39;s reagent), N-succinimidyl-S-acetylthioacetate (SATA), and SATP (N-succinimidyl-S-acetylthiopropionate). When the antibody is modified with acetylated reagents like SATA and SATP, acetyl groups can be removed via hydrolysis with hydroxylamine or similar reagents in order to generate free thiol groups for further conjugation. See, e.g., Traut et al. ( Biochem.,  12(17): 3266-3273 (1973)) and Duncan et al. ( Anal. Biochem.,  132(1): 68-73 (1983)). 
     For example, L M  can be of formula: 
     
       
         
         
             
             
         
       
     
     where L M  is bound to one or more thiol groups in an antibody, and the one or more thiol groups are naturally-occurring, solvent-accessible cysteine residues in the antibody, present in engineered cysteine residues, generated via full or partial reduction of disulfide linkages between cysteine sidechains in an antibody, or appended to lysine sidechains. When the wavy line (“ ”) crosses multiple bonds, it will be understood that the L M  attaches to the antibody at one or more positions (e.g., thiol groups). 
     The linking moiety can be derived from N-hydroxysuccinimidyl (NHS) esters or N-hydroxysulfosuccinimidyl (sulfo-NHS) esters; carbodiimides; hydroxymethyl phosphines; aryl azides; pentafluorophenyl (PFP) esters, tetrafluorophenyl (TFP) esters, or derivatives thereof; imidoesters; or vinyl sulfones, such that the linking moiety is attached to a free amine of the antibody. The free amine can be present in naturally-occurring solvent-accessible lysine residues in the antibody, engineered to be included in a non-naturally occurring lysine residue, or appended to a natural or unnatural amino acid residue of the antibody. 
     For example, L M  can be of formula: 
     
       
         
         
             
             
         
       
     
     where L M  is bound to one or more amine groups in an antibody, and the one or more amine groups are naturally-occurring solvent-accessible lysine residues in the antibody, engineered to be included in a non-naturally occurring lysine residue, or appended to a natural or unnatural amino acid residue of the antibody. 
     In some embodiments, the antibody can be modified to include an appended aldehyde, ketone, azide, or alkyne, such that the adjuvant can be linked via the aldehyde, ketone, azide, or alkyne. For example, L M  can be of formula: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     wherein the circle represents a 6 to 10-membered cyclic alkyl structure with the bond representing an attachment to one of the carbon atoms and when the wavy line (“ ”) crosses multiple bonds, it will be understood that the L M  attaches to the antibody at one or more positions (e.g., thiol groups). 
     In some embodiments, the adjuvant is attached to a cysteine residue with the thiol eliminated to form a dehydroalanine residue of the formula: 
     
       
         
         
             
             
         
       
     
     In such instances, L M  can be —S—, —NH—, or a bond. 
     The adjuvant can be linked to one or more naturally-occurring solvent-accessible tyrosine residues in the antibody or an engineered non-naturally occurring tyrosine residue such that, for example, L M  can be of formula: 
     
       
         
         
             
             
         
       
     
     wherein Tyr is a natural or an engineered non-naturally occurring tyrosine residue of the antibody (“Ab”). 
     In certain embodiments, L M  is a linking moiety of formula: 
     
       
         
         
             
             
         
       
     
     Immunoconjugates as described herein can provide an unexpectedly increased activation response of an antigen presenting cell (“APC”). This increased activation can be detected in vitro or in vivo. In some embodiments, the increased APC activation can be detected in the form of a reduced time to achieve a specified level of APC activation. For example, in an in vitro assay, % APC activation can be achieved at an equivalent dose with an immunoconjugate within about 1%, about 10%, about 20%, about 30%, about 40%, or about 50% of the time required to obtain the same or similar percentage of APC activation with a mixture of unconjugated antibody construct and adjuvant, under otherwise identical concentrations and conditions. In some embodiments, an immunoconjugate can activate APCs (e.g., dendritic cells) and/or NK cells in a reduced amount of time. For example, in some embodiments, a mixture of unconjugated antibody construct and adjuvant can activate APCs (e.g., dendritic cells) and/or NK cells and/or induce dendritic cell differentiation after incubation with the mixture for 2, 3, 4, 5, 1-5, 2-5, 3-5, or 4-7 days, while, in contrast, immunoconjugates described herein can activate and/or induce differentiation within 4 hours, 8 hours, 12 hours, 16 hours, or 1 day, under otherwise identical concentrations and conditions. Alternatively, the increased APC activation can be detected in the form of a reduced concentration of immunoconjugate required to achieve an amount (e.g., percent APCs), level (e.g., as measured by a level of upregulation of a suitable marker) or rate (e.g., as detected by a time of incubation required to activate) of APC activation. 
     In some embodiments, the immunoconjugates of the invention provide more than an about 5% increase in activity compared to a mixture of unconjugated antibody construct and adjuvant, under otherwise identical conditions. In other embodiments, the immunoconjugates of the invention provide more than an about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, or about 70% increase in activity compared to a mixture of unconjugated antibody construct and adjuvant, under otherwise identical conditions. The increase in activity can be assessed by any suitable means, many of which are known to those ordinarily skilled in the art and can include myeloid activation, assessment by cytokine secretion, or a combination thereof 
     In some embodiments, the invention provides an immunoconjugate of formula: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein subscript r is an integer from 1 to 10 and “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”). “Ab” can be any suitable antibody construct that has an antigen binding domain that binds HER2, such as, for example, trastuzumab and pertuzumab. In certain embodiments, “Ab” is trastuzumab (also known as HERCEPTIN™), a biosimilar thereof, or a biobetter thereof. For example, “Ab” can be MYL-14010, ABP 980, BCD-022, CT-P6, EG12014, HD201, ONS-1050, PF-05280014, Ontruzant, Saiputing, Herzuma, or HLX02. In preferred embodiments, “Ab” is trastuzumab (also known as HERCEPTIN™). 
     In some embodiments, the invention provides an immunoconjugate quaternary ammonium salt of formula: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     wherein counterion X −  is any pharmaceutically acceptable counterion (e.g., chloride, bromide, acetate, formate, nitrate, phosphate, sulfate, tosylate, etc.), subscript r is an integer from 1 to 10 and “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”). “Ab” can be any suitable antibody construct that has an antigen binding domain that binds HER2, such as, for example, trastuzumab and pertuzumab. In certain embodiments, “Ab” is trastuzumab (also known as HERCEPTIN™), a biosimilar thereof, or a biobetter thereof. For example, “Ab” can be MYL-14010, ABP 980, BCD-022, CT-P6, EG12014, HD201, ONS-1050, PF-05280014, Ontruzant, Saiputing, Herzuma, or HLX02. In preferred embodiments, “Ab” is trastuzumab (also known as HERCEPTIN™). 
     In some embodiments, the immunoconjugate is not of formula: 
     
       
         
         
             
             
         
       
         
         
           
             or a pharmaceutically acceptable salt thereof, wherein 
             Ab is an antibody; 
             T 1  is selected from C 1-6  alkyl and 2-to 6-membered heteroalkyl, each of which is optionally substituted with one or more members selected from the group consisting of halo, hydroxy, amino, oxo (═O), alkylamino, amido, acyl, nitro, cyano, and alkoxy; 
             T 2  is selected from O and CH 2 ; 
             each T 3  is independently CHT 6 , wherein T 6  is selected from H, OH, and NH 2 , 
             T 5  is a linker; 
             T 4  is selected from H and C 1-4  alkyl; or 
             T 5 , T 4 , and the nitrogen atom to which they are attached form a linker comprising a 5-to 8-membered heterocycle; 
             T 6  is an unmodified amino acid sidechain in the antibody or a modified amino acid sidechain in the antibody; 
             subscript n′ is an integer from 1 to 12 (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12); and 
             subscript r′ is an integer from 1 to 10 (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). 
           
         
       
    
     In some embodiments, the immunoconjugate is not of formula: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     wherein subscript r is an integer from 1 to 10 and “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”). 
     Adjuvants 
     The immunoconjugate of the invention comprises an adjuvant moiety. The adjuvant moiety described herein is a compound that elicits an immune response (i.e., an immunostimulatory agent). Generally, the adjuvant moiety described herein is a TLR agonist. TLRs are type-I transmembrane proteins that are responsible for the initiation of innate immune responses in vertebrates. TLRs recognize a variety of pathogen-associated molecular patterns from bacteria, viruses, and fungi and act as a first line of defense against invading pathogens. TLRs elicit overlapping yet distinct biological responses due to differences in cellular expression and in the signaling pathways that they initiate. Once engaged (e.g., by a natural stimulus or a synthetic TLR agonist), TLRs initiate a signal transduction cascade leading to activation of nuclear factor-κB (NF-κB) via the adapter protein myeloid differentiation primary response gene 88 (MyD88) and recruitment of the IL-1 receptor associated kinase (IRAK). Phosphorylation of IRAK then leads to recruitment of TNF-receptor associated factor 6 (TRAF6), which results in the 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 modes of regulation for TLR signaling include TIR-domain containing adapter-inducing interferon-β (TRIF)-dependent induction of TNF-receptor associated factor 6 (TRAF6) and activation of MyD88 independent pathways via TRIF and TRAF3, leading to the phosphorylation of interferon response factor three (IRF3). Similarly, the MyD88 dependent pathway also activates several IRF family members, including IRF5 and IRF7 whereas the TRIF dependent pathway also activates the NF-κB pathway. 
     Typically, the adjuvant moiety described herein is a TLR7 and/or TLR8 agonist. TLR7 and TLR8 are both expressed in monocytes and dendritic cells. In humans, TLR7 is also expressed in plasmacytoid dendritic cells (pDCs) and B cells. TLR8 is expressed mostly in cells of myeloid origin, i.e., monocytes, granulocytes, and myeloid dendritic cells. TLR7 and TLR8 are capable of detecting the presence of “foreign” single-stranded RNA within a cell, as a means to respond to viral invasion. Treatment of TLR8-expressing cells, with TLR8 agonists can result in 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 production of high levels of IFN-α and other inflammatory cytokines. TLR7/TLR8 engagement and resulting cytokine production can activate dendritic cells and other antigen-presenting cells, driving diverse innate and acquired immune response mechanisms leading to tumor destruction. 
     Antigen Binding Domain and Fc Domain 
     The immunoconjugates of the invention comprise an antibody construct that comprises an antigen binding domain that binds HER2. In some embodiments, the antibody construct further comprises an Fc domain. In certain embodiments, the antibody construct is an antibody. In certain embodiments, the antibody construct is a fusion protein. 
     The antigen binding domain can be a single-chain variable region fragment (scFv). A single-chain variable region fragment (scFv), which is a truncated Fab fragment including the variable (V) domain of an antibody heavy chain linked to a V domain of a light antibody chain via a synthetic peptide, can be generated using routine recombinant DNA technology techniques. Similarly, disulfide-stabilized variable region fragments (dsFv) can be prepared by recombinant DNA technology. 
     An embodiment of the invention provides antibody construct or antigen binding domain which specifically recognizes and binds to HER2 (SEQ ID NO: 1). The antibody construct or antigen binding domain may comprise one or more variable regions (e.g., two variable regions) of an antigen binding domain of an anti-HER2 antibody, each variable region comprising a CDR1, a CDR2, and a CDR3. 
     An embodiment of the invention provides an antibody construct or antigen binding domain comprising the CDR regions of trastuzumab. In this regard, the antibody construct or antigen binding domain may comprise a first variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO: 2 (CDR1 of first variable region), a CDR2 comprising the amino acid sequence of SEQ ID NO: 3 (CDR2 of first variable region), and a CDR3 comprising the amino acid sequence of SEQ ID NO: 4 (CDR3 of first variable region), and a second variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO: 5 (CDR1 of second variable region), a CDR2 comprising the amino acid sequence of SEQ ID NO: 6 (CDR2 of second variable region), and a CDR3 comprising the amino acid sequence of SEQ ID NO: 7 (CDR3 of second variable region). In this regard, the antibody construct can comprise (i) all of SEQ ID NOs: 2-4, (ii) all of SEQ ID NOs: 5-7, or (iii) all of SEQ ID NOs: 2-7. Preferably, the antibody construct or antigen binding domain comprises all of SEQ ID NOs: 2-7. 
     In an embodiment of the invention, the antibody construct or antigen binding domain comprising the CDR regions of trastuzumab further comprises the framework regions of the trastuzumab. In this regard, the antibody construct or antigen binding domain comprising the CDR regions of the trastuzumab further comprises the amino acid sequence of SEQ ID NO: 8 (framework region (“FR”) 1 of first variable region), the amino acid sequence of SEQ ID NO: 9 (FR2 of first variable region), the amino acid sequence of SEQ ID NO: 10 (FR3 of first variable region), the amino acid sequence of SEQ ID NO: 11 (FR4 of first variable region), the amino acid sequence of SEQ ID NO: 12 (FR1 of second variable region), the amino acid sequence of SEQ ID NO: 13 (FR2 of second variable region), the amino acid sequence of SEQ ID NO: 14 (FR3 of second variable region), and the amino acid sequence of SEQ ID NO: 15 (FR4 of second variable region). In this regard, the antibody construct or antigen binding domain can comprise (i) all of SEQ ID NOs: 2-4 and 8-11, (ii) all of SEQ ID NOs: 5-7 and 12-15; or (iii) all of SEQ ID NOs: 2-7 and 8-15. 
     An embodiment of the invention provides an antibody construct or antigen binding domain comprising one or both variable regions of trastuzumab. In this regard, the first variable region may comprise SEQ ID NO: 16. The second variable region may comprise SEQ ID NO: 17. Accordingly, in an embodiment of the invention, the antibody construct or antigen binding domain comprises SEQ ID NO: 16, SEQ ID NO: 17, or both SEQ ID NOs: 16 and 17. Preferably, the polypeptide comprises both of SEQ ID NOs: 16-17. 
     An embodiment of the invention provides an antibody construct or antigen binding domain comprising the CDR regions of pertuzumab. In this regard, the antibody construct or antigen binding domain may comprise a first variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO: 18 (CDR1 of first variable region), a CDR2 comprising the amino acid sequence of SEQ ID NO: 19 (CDR2 of first variable region), and a CDR3 comprising the amino acid sequence of SEQ ID NO: 20 (CDR3 of first variable region), and a second variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO: 21 (CDR1 of second variable region), a CDR2 comprising the amino acid sequence of SEQ ID NO: 22 (CDR2 of second variable region), and a CDR3 comprising the amino acid sequence of SEQ ID NO: 23 (CDR3 of second variable region). In this regard, the antibody construct can comprise (i) all of SEQ ID NOs: 18-20, (ii) all of SEQ ID NOs: 21-23, or (iii) all of SEQ ID NOs: 18-23. Preferably, the antibody construct or antigen binding domain comprises all of SEQ ID NOs: 18-23. 
     In an embodiment of the invention, the antibody construct or antigen binding domain comprising the CDR regions of pertuzumab further comprises the framework regions of the pertuzumab. In this regard, the antibody construct or antigen binding domain comprising the CDR regions of the pertuzumab further comprises the amino acid sequence of SEQ ID NO: 24 (framework region (“FR”) 1 of first variable region), the amino acid sequence of SEQ ID NO: 25 (FR2 of first variable region), the amino acid sequence of SEQ ID NO: 26 (FR3 of first variable region), the amino acid sequence of SEQ ID NO: 27 (FR4 of first variable region), the amino acid sequence of SEQ ID NO: 28 (FR1 of second variable region), the amino acid sequence of SEQ ID NO: 29 (FR2 of second variable region), the amino acid sequence of SEQ ID NO: 30 (FR3 of second variable region), and the amino acid sequence of SEQ ID NO: 31 (FR4 of second variable region). In this regard, the antibody construct or antigen binding domain can comprise (i) all of SEQ ID NOs: 18-20 and 24-26, (ii) all of SEQ ID NOs: 21-23 and 27-31; or (iii) all of SEQ ID NOs: 18-21 and 24-31. 
     An embodiment of the invention provides an antibody construct or antigen binding domain comprising one or both variable regions of pertuzumab. In this regard, the first variable region may comprise SEQ ID NO: 32. The second variable region may comprise SEQ ID NO: 33. Accordingly, in an embodiment of the invention, the antibody construct or antigen binding domain comprises SEQ ID NO: 32, SEQ ID NO: 33, or both SEQ ID NOs: 32 and 33. Preferably, the polypeptide comprises both of SEQ ID NOs: 32-33. 
     Included in the scope of the embodiments of the invention are functional variants of the antibody constructs or antigen binding domain described herein. The term “functional variant” as used herein refers to an antibody construct having an antigen binding domain with substantial or significant sequence identity or similarity to a parent antibody construct or antigen binding domain, which functional variant retains the biological activity of the antibody construct or antigen binding domain of which it is a variant. Functional variants encompass, for example, those variants of the antibody constructs or antigen binding domain described herein (the parent antibody construct or antigen binding domain) that retain the ability to recognize target cells expressing HER2 to a similar extent, the same extent, or to a higher extent, as the parent antibody construct or antigen binding domain. 
     In reference to the antibody construct or antigen binding domain, the functional variant can, for instance, be at least about 30%, about 50%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more identical in amino acid sequence to the antibody construct or antigen binding domain. 
     A functional variant can, for example, comprise the amino acid sequence of the parent antibody construct or antigen binding domain with at least one conservative amino acid substitution. Alternatively, or additionally, the functional variants can comprise the amino acid sequence of the parent antibody construct or antigen binding domain with at least one non-conservative amino acid substitution. In this case, it is preferable for the non-conservative amino acid substitution to not interfere with or inhibit the biological activity of the functional variant. The non-conservative amino acid substitution may enhance the biological activity of the functional variant, such that the biological activity of the functional variant is increased as compared to the parent antibody construct or antigen binding domain. 
     Amino acid substitutions of the inventive antibody constructs or antigen binding domains are preferably conservative amino acid substitutions. Conservative amino acid substitutions are known in the art, and include amino acid substitutions in which one amino acid having certain physical and/or chemical properties is exchanged for another amino acid that has the same or similar chemical or physical properties. For instance, the conservative amino acid substitution can be an acidic/negatively charged polar amino acid substituted for another acidic/negatively charged polar amino acid (e.g., Asp or Glu), an amino acid with a nonpolar side chain substituted for another amino acid with a nonpolar side chain (e.g., Ala, Gly, Val, Ile, Leu, Met, Phe, Pro, Trp, Cys, Val, etc.), a basic/positively charged polar amino acid substituted for another basic/positively charged polar amino acid (e.g., Lys, His, Arg, etc.), an uncharged amino acid with a polar side chain substituted for another uncharged amino acid with a polar side chain (e.g., Asn, Gln, Ser, Thr, Tyr, etc.), an amino acid with a beta-branched side-chain substituted for another amino acid with a beta-branched side-chain (e.g., Ile, Thr, and Val), an amino acid with an aromatic side-chain substituted for another amino acid with an aromatic side chain (e.g., His, Phe, Trp, and Tyr), etc. 
     The antibody construct or antigen binding domain can consist essentially of the specified amino acid sequence or sequences described herein, such that other components, e.g., other amino acids, do not materially change the biological activity of the antibody construct or antigen binding domain functional variant. 
     The antibody constructs and antigen binding domains of embodiments of the invention (including functional portions and functional variants) can be of any length, i.e., can comprise any number of amino acids, provided that the antibody constructs (or functional portions or functional variants thereof) retain their biological activity, e.g., the ability to specifically bind to HER2, detect cancer cells in a mammal, or treat or prevent cancer in a mammal, etc. For example, the antibody construct or antigen binding domain can be about 50 to about 5,000 amino acids long, such as 50, 70, 75, 100, 125, 150, 175, 200, 300, 400, 500, 600, 700, 800, 900, 1,000, or more amino acids in length. 
     The antibody constructs and antigen binding domains of embodiments of the invention (including functional portions and functional variants of the invention) can comprise synthetic amino acids in place of one or more naturally-occurring amino acids. Such synthetic amino acids are known in the art, and include, for example, aminocyclohexane carboxylic acid, norleucine, α-amino n-decanoic acid, homoserine, S-acetylaminomethyl-cysteine, trans-3- and trans-4-hydroxyproline, 4-aminophenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenylalanine, β-phenylserine β-hydroxyphenylalanine, phenylglycine, α-naphthylalanine, cyclohexylalanine, cyclohexylglycine, indoline-2-carboxylic acid, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, aminomalonic acid, aminomalonic acid monoamide, N′-benzyl-N′-methyl-lysine, N′,N′-dibenzyl-lysine, 6-hydroxylysine, ornithine, α-aminocyclopentane carboxylic acid, α-aminocyclohexane carboxylic acid, α-aminocycloheptane carboxylic acid, α-(2-amino-2-norbornane)-carboxylic acid, α,γ-diaminobutyric acid, α,β-diaminopropionic acid, homophenylalanine, and α-tert-butylglycine. 
     The antibody constructs of embodiments of the invention (including functional portions and functional variants) can be glycosylated, amidated, carboxylated, phosphorylated, esterified, N-acylated, cyclized via, e.g., a disulfide bridge, or converted into an acid addition salt and/or optionally dimerized or polymerized. 
     In some embodiments, the antibody construct is a monoclonal antibody of a defined sub-class (e.g., IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 , or IgA 2 ). If combinations of antibodies are used, the antibodies can be from the same subclass or from different subclasses. Typically, the antibody construct is an IgG 1  antibody. Various combinations of different subclasses, in different relative proportions, can be obtained by those of skill in the art. In some embodiments, a specific subclass or a specific combination of different subclasses can be particularly effective at cancer treatment or tumor size reduction. Accordingly, some embodiments of the invention provide immunoconjugates wherein the antibody is a monoclonal antibody. In some embodiments, the monoclonal antibody is a humanized monoclonal antibody. 
     In some embodiments, the antibody construct or antigen binding domain binds to HER2 on a cancer or immune cell at a higher affinity than a corresponding HER2 antigen on a non-cancer cell. For example, the antibody construct or antigen binding domain may preferentially recognize HER2 containing a polymorphism that is found on a cancer or immune cell as compared to recognition of a corresponding wild-type HER2 antigen on the non-cancer. In some embodiments, the antibody construct or antigen binding domain binds a cancer cell with greater avidity than a non-cancer cell. For example, the cancer cell can express a higher density of HER2, thereby providing for a higher affinity binding of a multivalent antibody to the cancer cell. 
     In some embodiments, the antibody construct or antigen binding domain does not significantly bind non-cancer antigens (e.g., the antibody binds one or more non-cancer antigens with at least 10, 100, 1,000, 10,000, 100,000, or 1,000,000-fold lower affinity (higher Kd) than HER2). In some embodiments, the corresponding non-cancer cell is a cell of the same tissue or origin that is not hyperproliferative or otherwise cancerous. HER2 need not be specific to the cancer cell or even enriched in cancer cells relative to other cells (e.g., HER2 can be expressed by other cells). Thus, in the phrase “an antibody construct that specifically binds to an antigen of a cancer cell,” the term “specifically” refers to the specificity of the antibody construct and not to the uniqueness of the presence of HER2 in that particular cell type. 
     Modified Fc Region 
     In some embodiments, the antibodies in the immunoconjugates contain a modified Fc region, wherein the modification modulates the binding of the Fc region to one or more Fc receptors. 
     The terms “Fc receptor” or “FcR” refer to a receptor that binds to the Fc region of an antibody. There are three main classes of Fc receptors: (1) FcγR which bind to IgG, (2) FcαR which binds to IgA, and (3) FcR which binds to IgE. The FcγR family includes several members, such as FcγI (CD64), FcγRIIA (CD32A), FcγRIIB (CD32B), FcγRIIIA (CD16A), and FcγRIIIB (CD16B). The Fcγ receptors differ in their affinity for IgG and also have different affinities for the IgG subclasses (e.g., IgG1, IgG2, IgG3, and IgG4). 
     In some embodiments, the antibodies in the immunoconjugates (e.g., antibodies conjugated to at least two adjuvant moieties) contain one or more modifications (e.g., amino acid insertion, deletion, and/or substitution) in the Fc region that results in modulated binding (e.g., increased binding or decreased binding) to one or more Fc receptors (e.g., FcγRI (CD64), FcγRIIA (CD32A), FcγRIIB (CD32B), FcγRIIIA (CD16a), and/or FcγRIIIB (CD16b)) as compared to the native antibody lacking the mutation in the Fc region. In some embodiments, the antibodies in the immunoconjugates contain one or more modifications (e.g., amino acid insertion, deletion, and/or substitution) in the Fc region that reduce the binding of the Fc region of the antibody to FcγRIIB. In some embodiments, the antibodies in the immunoconjugates contain one or more modifications (e.g., amino acid insertion, deletion, and/or substitution) in the Fc region of the antibody that reduce the binding of the antibody to FcγRIIB while maintaining the same binding or having increased binding to FcγRI (CD64), FcγRIIA (CD32A), and/or FcRγIIIA (CD16a) as compared to the native antibody lacking the mutation in the Fc region. In some embodiments, the antibodies in the immunoconjugates contain one of more modifications in the Fc region that increase the binding of the Fc region of the antibody to FcγRIIB. 
     In some embodiments, the modulated binding is provided by mutations in the Fc region of the antibody relative to the native Fc region of the antibody. The mutations can be in a CH2 domain, a CH3 domain, or a combination thereof. A “native Fc region” is synonymous with a “wild-type Fc region” and comprises an amino acid sequence that is identical to the amino acid sequence of an Fc region found in nature or identical to the amino acid sequence of the Fc region found in the native antibody (e.g., trastuzumab, pertuzumab, biosimilars thereof, or biobetters thereof). Native sequence human Fc regions include a native sequence human IgG1 Fc region, native sequence human IgG2 Fc region, native sequence human IgG3 Fc region, and native sequence human IgG4 Fc region, as well as naturally occurring variants thereof. Native sequence Fc includes the various allotypes of Fcs (see, e.g., Jefferis et al.,  mAbs,  1(4): 332-338 (2009)). 
     In some embodiments, the mutations in the Fc region that result in modulated binding to one or more Fc receptors can include one or more of 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 V11 (G237D/P238D/H268D/P271G/A330R), and/or one or more mutations at the following amino acids: E233, G237, P238, H268, P271, L328 and A330. Additional Fc region modifications for modulating Fc receptor binding are described in, for example, U.S. Patent Application Publication 2016/0145350 and U.S. Pat. Nos. 7,416,726 and 5,624,821, which are hereby incorporated by reference in their entireties. 
     In some embodiments, the Fc region of the antibodies of the immunoconjugates are modified to have an altered glycosylation pattern of the Fc region compared to the native non-modified Fc region. 
     Human immunoglobulin is glycosylated at the Asn297 residue in the Cγ2 domain of each heavy chain. This N-linked oligosaccharide is composed of a core heptasaccharide, N-acetylglucosamine4Mannose3 (GlcNAc4Man3). Removal of the heptasaccharide with endoglycosidase or PNGase F is known to lead to conformational changes in the antibody Fc region, which can significantly reduce antibody-binding affinity to activating FcγR and lead to decreased effector function. The core heptasaccharide is often decorated with galactose, bisecting GlcNAc, fucose, or sialic acid, which differentially impacts Fc binding to activating and inhibitory FcγR. Additionally, it has been demonstrated that α2,6-sialyation enhances anti-inflammatory activity in vivo, while defucosylation leads to improved FcγRIIIa binding and a 10-fold increase in antibody-dependent cellular cytotoxicity and antibody-dependent phagocytosis. Specific glycosylation patterns, therefore, can be used to control inflammatory effector functions. 
     In some embodiments, the modification to alter the glycosylation pattern is a mutation. For example, a substitution at Asn297. In some embodiments, Asn297 is mutated to glutamine (N297Q). Methods for controlling immune response with antibodies that modulate FcγR-regulated signaling are described, for example, in U.S. Pat. No. 7,416,726 and U.S. Patent Application Publications 2007/0014795 and 2008/0286819, which are hereby incorporated by reference in their entireties. 
     In some embodiments, the antibodies of the immunoconjugates are modified to contain an engineered Fab region with a non-naturally occurring glycosylation pattern. For example, hybridomas can be genetically engineered to secrete afucosylated mAb, desialylated mAb or deglycosylated Fc with specific mutations that enable increased FcRγIIIa binding and effector function. In some embodiments, the antibodies of the immunoconjugates are engineered to be afucosylated. 
     In some embodiments, the entire Fc region of an antibody construct in the immunoconjugates is exchanged with a different Fc region, so that the Fab region of the antibody is conjugated to a non-native Fc region. For example, the Fab region of trastuzumab, which normally comprises an IgG1 Fc region, can be conjugated to IgG2, IgG3, IgG4, or IgA, or the Fab region of nivolumab, which normally comprises an IgG4 Fc region, can be conjugated to IgG1, IgG2, IgG3, IgA1, or IgG2. In some embodiments, the Fc modified antibody with a non-native Fc domain also comprises one or more amino acid modification, such as the S228P mutation within the IgG4 Fc, that modulate the stability of the Fc domain described. In some embodiments, the Fc modified antibody with a non-native Fc domain also comprises one or more amino acid modifications described herein that modulate Fc binding to FcR. 
     In some embodiments, the modifications that modulate the binding of the Fc region to FcR do not alter the binding of the Fab region of the antibody to its antigen when compared to the native non-modified antibody. In other embodiments, the 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 the native non-modified antibody. 
     Immunoconjugate Composition 
     The invention provides a composition, e.g., a pharmaceutically acceptable composition or formulation, comprising a plurality of immunoconjugates as described herein and optionally a carrier therefor, e.g., a pharmaceutically acceptable carrier. The immunoconjugates can be the same or different in the composition, i.e., the composition can comprise immunoconjugates that have the same number of adjuvants linked to the same positions on the antibody construct and/or immunoconjugates that have the same number of adjuvants linked to different positions on the antibody construct, that have different numbers of adjuvants linked to the same positions on the antibody construct, or that have different numbers of adjuvants linked to different positions on the antibody construct. 
     A composition of immunoconjugates of the invention can have an average adjuvant to antibody construct ratio of about 0.4, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, 3, 3.2, 3.4, 3.6, 3.8, 4.0, 4.2, 4.4, 4.6, 4.8, 5.0, 5.2, 5.4, 5.6, 5.8, 6.0, 6.2, 6.4, 6.6, 6.8, 7, 7.2, 7.4, 7.6, 7.8, 8, 8.2, 8.4, 8.6, 8.8, 9, 9.2, 9.4, 9.6, 9.8, or 10, or within a range bounded by any two of the aforementioned values. A skilled artisan will recognize that the number of adjuvant conjugated to the antibody construct may vary from immunoconjugate to immunoconjugate in a composition comprising multiple immunoconjugates of the invention, and, thus, the adjuvant to antibody construct (e.g., antibody) ratio can be measured as an average. The adjuvant to antibody construct (e.g., antibody) ratio can be assessed by any suitable means, many of which are known in the art. 
     In some embodiments, the composition further comprises one or more pharmaceutically 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 immunoconjugates can be injected intra-tumorally. Compositions for injection will commonly comprise a solution of the immunoconjugate dissolved in a pharmaceutically acceptable carrier. Among the acceptable vehicles and solvents that can be employed are water and an isotonic solution of one or more salts such as sodium chloride, e.g., Ringer&#39;s solution. In addition, sterile fixed oils can conventionally be employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can likewise be used in the preparation of injectables. These compositions desirably are sterile and generally free of undesirable matter. These compositions can be sterilized by conventional, well known sterilization techniques. The compositions can contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents, e.g., sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. 
     The composition can contain any suitable concentration of the immunoconjugate. The concentration of the immunoconjugate in the composition can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight, and the like, in accordance with the particular mode of administration selected and the patient&#39;s needs. In certain embodiments, the concentration of an immunoconjugate in a solution formulation for injection will range from about 0.1% (w/w) to about 10% (w/w). 
     Methods of Using the Immunoconjugate 
     The invention provides a method for treating cancer. The method includes administering a therapeutically effective amount of an immunoconjugate as described herein (e.g., as a composition as described herein) to a subject in need thereof, e.g., a subject that has cancer and is in need of treatment for the cancer. 
     Trastuzumab, pertuzumab, biosimilars thereof, and biobetters thereof are known to be useful in the treatment of cancer, particularly breast cancer, especially HER2-overexpressing breast cancer, gastric cancer, especially HER2-overexpressing gastric cancer, and gastroesophageal junction adenocarcinoma. The immunoconjugate described herein can be used to treat the same types of cancers as trastuzumab, pertuzumab, biosimilars thereof, and biobetters thereof, particularly breast cancer, especially HER2-overexpressing breast cancer, gastric cancer, especially HER2-overexpressing gastric cancer, and gastroesophageal junction adenocarcinoma. 
     The immunoconjugate is administered to a subject in need thereof in any therapeutically effective amount using any suitable dosing regimen, such as the dosing regimens utilized for trastuzumab, pertuzumab, biosimilars thereof, and biobetters thereof. For example, the methods can include administering the immunoconjugate to provide a dose of from about 100 ng/kg to about 50 mg/kg to the subject. The immunoconjugate dose can range from about 5 mg/kg to about 50 mg/kg, from about 10 μg/kg to about 5 mg/kg, or from 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. The immunoconjugate dose can also be outside of these ranges, depending on the particular conjugate as well as the type and severity of the cancer being treated. 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 once per month to about five times per week. In some embodiments, the immunoconjugate is administered once per week. 
     In another aspect, the invention provides a method for preventing cancer. The method comprises administering a therapeutically effective amount of an immunoconjugate (e.g., as a composition as described above) to a subject. In certain embodiments, the subject is susceptible to a certain cancer to be prevented. For example, the methods can include administering the immunoconjugate to provide a dose of from about 100 ng/kg to about 50 mg/kg to the subject. The immunoconjugate dose can range from about 5 mg/kg to about 50 mg/kg, from about 10 μg/kg to about 5 mg/kg, or from 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. The immunoconjugate dose can also be outside of these ranges, depending on the particular conjugate as well as the type and severity of the cancer being treated. 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 once per month to about five times per week. In some embodiments, the immunoconjugate is administered once per week. 
     Some embodiments of the invention provide methods for treating cancer as described above, wherein the cancer is breast cancer. Breast cancer can originate from different areas in the breast, and a number of different types of breast cancer have been characterized. For example, the immunoconjugates of the invention can be used for treating ductal carcinoma in situ; invasive ductal carcinoma (e.g., tubular carcinoma; medullary carcinoma; mucinous carcinoma; papillary carcinoma; or cribriform carcinoma of the breast); lobular carcinoma in situ; invasive lobular carcinoma; inflammatory breast cancer; and other forms of breast cancer. In some embodiments, methods for treating breast cancer include administering an immunoconjugate containing an antibody construct that is capable of binding HER2 (e.g., trastuzumab, pertuzumab, biosimilars thereof, or biobetters thereof). 
     Some embodiments of the invention provide methods for treating cancer as described above, wherein the cancer is gastric cancer. Gastric (stomach) cancer can originate from different cells in the stomach and several types of gastric cancer have been characterized including adenocarcinoma, carcinoid tumors, squamous cell carcinoma, small cell carcinoma, leiomyosarcoma, and gastrointestinal stromal tumors. In some embodiments, methods for treating gastric cancer include administering an immunoconjugate containing an antibody construct that is capable of binding HER2 (e.g., trastuzumab, pertuzumab, biosimilars thereof, or biobetters thereof). 
     Some embodiments of the invention provide methods for treating cancer as described above, wherein the cancer is gastroesophageal junction carcinoma. This carcinoma occurs in the area where the esophagus meats the stomach. There are three types of gastroesophageal junction carcinoma. In Type 1, the cancer the cancer grows down from above and into the gastroesophageal junction. The normal lining of the lower end of the esophagus is replaced by mutations (also called Barrett&#39;s esophagus). In Type 2, the cancer grows at the gastroesophageal junction by itself. In Type 3, the cancer grows up into the gastroesophageal junction from the stomach upwards. In some embodiments, methods for treating gastroesophageal junction carcinoma include administering an immunoconjugate containing an antibody construct that is capable of binding HER2 (e.g., trastuzumab, pertuzumab, biosimilars thereof, or biobetters thereof). 
     In some embodiments, the cancer is susceptible to a pro-inflammatory response induced by TLR7 and/or TLR8. 
     EXAMPLES OF NON-LIMITING ASPECTS OF THE DISCLOSURE 
     Aspects, including embodiments, of the invention described herein may be beneficial alone or in combination, with one or more other aspects or embodiments. Without limiting the foregoing description, certain non-limiting aspects of the disclosure numbered 1-26 are provided below. As will be apparent to those of skill in the art upon reading this disclosure, each of the individually numbered aspects may be used or combined with any of the preceding or following individually numbered aspects. This is intended to provide support for all such combinations of aspects and is not limited to combinations of aspects explicitly provided below: 
     1. An immunoconjugate of formula (I) or formula (II): 
     
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, 
     wherein 
     R 1  and R 2  independently are hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     J 1  is CH or N, 
     J 2  is CHQ, NQ, O, or S, 
     each Q independently is Y or Z, wherein exactly one Q is Y, 
     Y is of formula: 
     
       
         
         
             
             
         
       
     
     each Z independently is hydrogen or of formula: 
     
       
         
         
             
             
         
       
     
     A is optionally present and is NR 6  or of formula: 
     
       
         
         
             
             
         
       
     
     U is optionally present and is CH 2 , C(O), CH 2 C(O), or C(O)CH 2 , 
     R 6  and W independently are hydrogen, Ar 1 , or of formula: 
     
       
         
         
             
             
         
       
     
     V is optionally present and is of formula: 
     
       
         
         
             
             
         
       
     
     J 3  and J 4  independently are CH or N, 
     m 1 , m 2 , and m 3  independently are an integer from 0 to 25, except that at least one of m 1 , m 2 , and m 3  is a non-zero integer, 
     n 1 , n 2 , n 3 , n 4 , n 5 , and n 6  independently are an integer from 0 to 10, 
     t 1  and t 2  independently are an integer from 1 to 3, 
     G 1 , G 2 , G 3 , and G 4  independently are CH 2 , C(O), CH 2 C(O), C(O)CH 2 , or a bond, 
     X 1 , X 2 , X 3 , and X 4  are each optionally present and independently are O, NR 9 , CHR 9 , SO 2 , S, or one or two divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, and when more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is present, the more than one divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked or fused, wherein linked divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups are linked through a bond or —CO—, 
     R 4  is hydrogen, C 1 -C 4  alkyl, 
     
       
         
         
             
             
         
       
     
     R 3 , R 5 , R 7 , R 8 , R 9 , R 10 , and R 11  independently are hydrogen or C 1 -C 4  alkyl, 
     Ar 1  and Ar 2  independently are an aryl or heteroaryl group, optionally substituted with one or more halogens (e.g., fluorine, chlorine, bromine, or iodine), nitriles, hydroxyls, C 1 -C 4  alkyl groups, or a combination thereof, 
     L M  is a linking moiety, 
     r is an integer from 1 to 10, 
     “Ab” is an antibody construct that has an antigen binding domain that binds human epidermal growth factor receptor 2 (“HER2”), and
         each wavy line (“ ”) represents a point of attachment.       

     2. The immunoconjugate of aspect 1, wherein subscript r is an integer from 1 to 6. 
     3. The immunoconjugate of aspect 2, wherein subscript r is an integer from 1 to 4. 
     4. The immunoconjugate of aspect 3, wherein subscript r is 1. 
     5. The immunoconjugate of aspect 3, wherein subscript r is 2. 
     6. The immunoconjugate of aspect 3, wherein subscript r is 3. 
     7. The immunoconjugate of aspect 3, wherein subscript r is 4. 
     8. The immunoconjugate of aspect 1, wherein the immunoconjugate is of formula: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof, wherein subscript r is an integer from 1 to 10 and “Ab” is an antibody construct that has an antigen binding domain that binds HER2. 
     9. The immunoconjugate of any one of aspects 1-8, wherein “Ab” is trastuzumab, a biosimilar thereof, or a biobetter thereof 
     10. The immunoconjugate of any one of aspects 1-8, wherein “Ab” is pertuzumab, a biosimilar thereof, or a biobetter thereof. 
     11. The immunoconjugate of aspect 9, wherein “Ab” is trastuzumab. 
     12. A composition comprising a plurality of immunoconjugates according to any one of aspects 1-11. 
     13. The composition of aspect 12, wherein the average adjuvant to antibody construct ratio is from about 0.01 to about 10. 
     14. The composition of aspect 13, wherein the average adjuvant to antibody construct ratio is from about 1 to about 10. 
     15. The composition of aspect 14, wherein the average adjuvant to antibody construct ratio is from about 1 to about 6. 
     16. The composition of aspect 15, wherein the average adjuvant to antibody construct ratio is from about 1 to about 4. 
     17. The composition of aspect 16, wherein the average adjuvant to antibody construct ratio is from about 1 to about 3. 
     18. A method for treating cancer comprising administering a therapeutically effective amount of an immunoconjugate according to any one of aspects 1-11 or a composition according to any one of aspects 12-17 to a subject in need thereof. 
     19. The method of aspect 18, wherein the cancer is susceptible to a pro-inflammatory response induced by TLR7 and/or TLR8 agonism. 
     20. The method of aspect 18 or 19, wherein the cancer is a HER2-expressing cancer. 
     21. The method of any one of aspects 18-20, wherein the cancer is breast cancer. 
     22. The method of aspect 21, wherein the breast cancer is HER2 overexpressing breast cancer. 
     23. The method of any one of aspects 18-20, wherein the cancer is gastric cancer. 
     24. The method of aspect 23, wherein the gastric cancer is HER2 overexpressing gastric cancer. 
     25. The method of any one of aspects 18-20, 23, and 24, wherein the cancer is gastroesophageal junction adenocarcinoma. 
     26. Use of an immunoconjugate according to any one of aspects 1-11 or a composition according to any one of aspects 12-17 for treating cancer. 
     EXAMPLES 
     The following examples further illustrate the invention but, of course, should not be construed as in any way limiting its scope. 
     Example 1: Synthesis of Compound 2 
     
       
         
         
             
             
         
       
     
     A mixture of 2,2-dimethyl-1,3-dioxane-4,6-dione (41.89 g, 290.66 mmol, 1 eq) and 4-bromoaniline (50 g, 290.66 mmol, 1 eq) was stirred (neat) at 80° C. for 12 hrs. Afterward, the small remaining amount of acetone was removed by vacuum. Eaton&#39;s reagent (415.15 g, 1.74 mol, 273.12 mL, 6 eq) was added to the mixture at 80° C. for 12 hrs. Water (1000 mL) was added to this mixture while stirring vigorously. The precipitate was filtered, washed with H 2 O, and air dried to provide a solid. The solid was recrystallized from ethanol to afford 6-bromoquinoline-2,4-diol (26 g, 108.31 mmol, 37.26% yield) as off-white solid.  1 H NMR (dimethyl sulfoxide (DMSO)-d 6 , 400 MHz) δ 11.53 (s, 1H), 11.33 (s, 1H), 7.85 (d, J=2.4 Hz, 1H), 7.75 (dd, J=8.0 Hz, 4.0 Hz, 1H), 7.18-7.24 (m, 1H), 5.75 (s, 1H). 
     Example 2: Synthesis of Compound 3 
     
       
         
         
             
             
         
       
     
     To a solution of nitric acid HNO 3  (13.65 g, 216.62 mmol, 9.75 mL, 2 eq) in AcOH (500 mL) was added 6-bromoquinoline-2,4-diol (26 g, 108.31 mmol, 1 eq) slowly at 15° C. The mixture was stirred at 80° C. for 3 hours. The mixture was cooled and quenched by addition of water (1000 mL). The product was separated by filtration and washed by water (100 mL×3), dried to give desired product. The crude product 6-bromo-3-nitro-quinoline-2,4-diol (30 g, 105.24 mmol, 97.17% yield) was obtained as a yellow solid and used into the next step without further purification.  1 H NMR (DMSO-d 6 , 400 MHz) δ 11.92 (s, 1H), 8.13 (s, 1H), 7.76 (d, J=8.4 Hz, 1H), 7.25 (d, J=8.4 Hz, 1H). 
     Example 3: Synthesis of Compound 4 
     
       
         
         
             
             
         
       
     
     To a mixture of 6-bromo-3-nitro-quinoline-2,4-diol (30 g, 105.24 mmol, 1 eq) in POCl 3  (484.12 g, 3.16 mol, 293.41 mL, 30 eq) was added N,N-diisopropylethylamine (40.81 g, 315.73 mmol, 55.00 mL, 3 eq) slowly at 15° C. The mixture was stirred at 100° C. for 16 hrs. The mixture was concentrated in vacuum. The residue was poured into ice water (2000 mL), filtered and washed with H 2 O (500 mL×3), and dried to provide 6-bromo-2,4-dichloro-3-nitro-quinoline (30 g, 93.18 mmol, 88.54% yield) as a yellow solid.  1 H NMR (DMSO-d 6 , 400 MHz) δ 8.48 (d, J=2.0 Hz, 1H), 8.25 (dd, J=8.8, 2.0 Hz, 1H), 8.10 (d, J=8.8 Hz, 1H). 
     Example 4: Synthesis of Compound 5 
     
       
         
         
             
             
         
       
     
     To a mixture of 6-bromo-2,4-dichloro-3-nitro-quinoline (10.00 g, 31.06 mmol, 1 eq) and 4-aminobutan-1-ol (2.77 g, 31.06 mmol, 2.89 mL, 1 eq) in THE (100 mL) was added Et 3 N (4.71 g, 46.59 mmol, 6.49 mL, 1.5 eq) in one portion at 0° C. The mixture was stirred at 0° C. for 2 hrs. The mixture was diluted with water (200 mL) and extracted with EtOAc(100 mL×3). The organic layer was washed with brine (100 mL), dried over Na 2 SO 4 , filtered and concentrated to provide 4-[(6-bromo-2-chloro-3-nitro-4-quinolyl)amino]butan-1-ol (12 g, crude) as a yellow solid.  1 H NMR (DMSO-d 6 , 400 MHz) δ 8.72-8.85 (m, 1H), 7.90-7.98 (m, 2H), 7.70-7.75 (m, 1H), 4.49 (t, J=3.2 Hz, 1H), 3.36-3.43 (m, 2H), 3.07-3.15 (m, 2H), 1.63-1.73 (m, 2H), 1.39-1.48 (m, 2H). 
     Example 5: Synthesis of Compound 6 
     
       
         
         
             
             
         
       
     
     To a mixture of 4-[(6-bromo-2-chloro-3-nitro-4-quinolyl)amino]butan-1-ol (12 g, 32.03 mmol, 1 eq) in THE (200 mL) was added imidazole (2.83 g, 41.64 mmol, 1.3 eq) and tert-butyldimethylsilyl chloride (6.28 g, 41.64 mmol, 5.10 mL, 1.3 eq) slowly in portions at 25° C. The mixture was stirred at 25° C. for 2 hrs. The mixture was diluted with water (200 mL) and extracted with EtOAc (200 mL×3). The organic layer was washed with brine (100 mL), dried over Na 2 SO 4 , filtered and concentrated. 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, 5/1). Compound 6-bromo-N-[4-[tert-butyl (dimethyl)silyl]oxybutyl]-2-chloro-3-nitro-quinolin-4-amine (15 g, 30.68 mmol, 95.79% yield) was obtained as a yellow solid.  1 H NMR (DMSO-d 6 , 400 MHz) δ 8.80 (d, J=2.0 Hz, 1H), 7.92-7.98 (m, 2H), 7.70-7.78 (m, 1H), 3.52-3.59 (m, 2H), 3.09-3.16 (m, 2H), 1.63-1.72 (m, 2H), 1.42-1.52 (m, 2H), 0.81 (s, 9H), −0.02 (s, 6H). 
     Example 6: Synthesis of Compound 7 
     
       
         
         
             
             
         
       
     
     To a solution of 6-bromo-N-[4-[tert-butyl(dimethyl)silyl]oxybutyl]-2-chloro-3-nitro-quinolin-4-amine (15 g, 30.68 mmol, 1 eq) in EtOAc (200 mL) was added platinum on carbon (3.87 g, 920.48 μmol, 5% purity, 0.03 eq) under N 2 . The suspension was degassed under vacuum and purged with H 2  several times. The mixture was stirred under H 2  (50 psi) at 25° C. for 12 hrs. The mixture was filtered and concentrated. Compound 6-bromo-N 4 -(4-((tert-butyldimethylsilyl)oxy)butyl)-2-chloroquinoline-3,4-diamine (13.6 g, 29.64 mmol, 96.59% yield) was obtained as a yellow oil.  1 H NMR (CDCl 3 , 400 MHz) δ 7.92 (d, J=2.0 Hz, 1H), 7.75 (d, J=9.2 Hz, 1H), 7.51-7.55 (m, 1H), 4.17-4.22 (m, 1H), 3.65-3.81 (m, 2H), 3.07-3.37 (m, 2H), 1.53-1.85 (m, 4H), 0.92 (s, 9H), 0.06 (s, 6H). 
     Example 7: Synthesis of Compound 8 
     
       
         
         
             
             
         
       
     
     To a mixture of 6-bromo-N 4 -[4-[tert-butyl(dimethyl)silyl]oxybutyl]-2-chloro-quinoline-3,4-diamine (13.6 g, 29.64 mmol, 1 eq) and pentanoyl chloride (6.07 g, 50.38 mmol, 6.11 mL, 1.7 eq) in THE (200 mL) was added Et 3 N (4.50 g, 44.45 mmol, 6.19 mL, 1.5 eq) in batches at 0° C. The mixture was stirred at 25° C. for 3 hrs. The mixture was diluted with water (200 mL) and extracted with EtOAc (100 mL×3). The organic layer was washed with brine (100 mL), dried over Na 2 SO 4 , filtered and concentrated. The residue was purified by flash silica gel chromatography (Teledyne Isco, Lincoln, Nebr.), 20 g SEPAFLASH™ silica flash column, eluent of 0 to about 90% ethyl acetate/petroleum ether gradient at 100 mL/min). Compound N-[6-bromo-4-[4-[tert-butyl (dimethyl) silyl]oxybutylamino]-2-chloro-3-quinolyl]pentanamide (9 g, 16.57 mmol, 55.93% yield) was obtained as a white solid.  1 H NMR (DMSO-d 6 , 400 MHz) δ 9.47 (s, 1H), 8.56 (d, J=2.0 Hz, 1H), 7.74-7.81 (m, 1H), 7.60-7.67 (m, 1H), 6.91-6.99 (m, 1H), 3.55-3.61 (m, 2H), 3.39-3.46 (m, 2H), 2.30-2.35 (m, 2H), 1.54-1.65 (m, 4H), 1.43-1.53 (m, 2H), 1.31-1.39 (m, 2H), 0.87-0.95 (m, 3H), 0.83 (s, 9H), 0.00 (s, 6H). 
     Example 8: Synthesis of Compound 9 
     
       
         
         
             
             
         
       
     
     To a mixture of N-[6-bromo-4-[4-[tert-butyl(dimethyl)silyl]oxybutylamino]-2-chloro-3-quinolyl]pentanamide (9 g, 16.57 mmol, 1 eq) in toluene (150 mL) was added AcOH (1.99 g, 33.15 mmol, 1.90 mL, 2 eq) in one portion at 25° C. The mixture was stirred at 100° C. for 12 hrs. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue mixture was washed with EtOAc (50 mL). The mixture was filtered, and the filtrate was concentrated under reduced pressure to provide a residue. The residue was purified by flash silica gel chromatography (Teledyne Isco, 7 g, SEPAFLASH™ silica flash column, eluent of 0 to 100% ethyl acetate/petroleum ether gradient at 100 mL/min). Compound 4-(8-bromo-2-butyl-4-chloro-imidazo[4,5-c]quinolin-1-yl)butoxy-tert-butyl-dimethyl-silane (5 g, 9.52 mmol, 57.46% yield) was obtained as a white solid.  1 H NMR (DMSO-d 6 , 400 MHz) δ 8.42 (d, J=4.0 Hz, 1H), 7.97-8.03 (m, 1H), 7.83-7.90 (m, 1H), 4.67 (t, J=8.0 Hz, 2H), 3.61 (t, J=4.0 Hz, 2H), 3.0 (t, J=8.0 Hz, 2H), 1.78-1.94 (m, 4H), 1.52-1.62 (m, 2H), 1.43-1.50 (m, 2H), 0.97 (t, J=8.0 Hz, 3H), 0.72 (s, 9H), −0.06 (s, 6H). 
     Example 9: Synthesis of Compound 10 
     
       
         
         
             
             
         
       
     
     To a mixture of 4-(8-bromo-2-butyl-4-chloro-imidazo[4,5-c]quinolin-1-yl)butoxy-tert-butyl-dimethyl-silane (6.2 g, 11.81 mmol, 1 eq) in (2,4-dimethoxyphenyl) methanamine (19.75 g, 118.10 mmol, 17.79 mL, 10 eq) at 20° C. The mixture was stirred at 120° C. for 3 hrs. The mixture was diluted with water (200 mL) and extracted with EtOAc (100 ml×3). The organic layer was washed with brine (100 mL), dried over Na 2 SO 4 , filtered and concentrated. The residue was purified by flash silica gel chromatography (Teledyne Isco, 10 g, SEPAFLASH™ silica flash column, eluent of 0 to about 50% ethyl acetate/petroleum ether gradient at 100 mL/min) to provide 8-bromo-2-butyl-1-[4-[tert-butyl(dimethyl)silyl]oxybutyl]-N-[(2,4-dimethoxyphenyl)methyl]imidazo[4,5-c]quinolin-4-amine (8.7 g, crude) as a yellow oil.  1 H NMR (DMSO-d 6 , 400 MHz) δ 7.99-8.07 (m, 1H), 7.52-7.57 (m, 1H), 7.44-7.51 (m, 1H), 7.10-7.17 (m, 1H), 6.98-7.06 (m, 1H), 6.56 (d, J=2.0 Hz, 1H), 6.39 (dd, J=8.0 Hz, 2.0 Hz, 1H), 4.63-4.69 (m, 2H), 4.45-4.53 (m, 2H), 3.81-3.85 (m, 3H), 3.67-3.73 (m, 3H), 3.56-3.66 (m, 2H), 2.85-2.95 (m, 2H), 1.74-1.90 (m, 4H), 1.51-1.63 (m, 2H), 1.37-1.48 (m, 2H), 0.89-0.98 (m, 3H), 0.76 (s, 9H), −0.04 (s, 6H). 
     Example 10: Synthesis of Compound 11 
     
       
         
         
             
             
         
       
     
     To a mixture of tert-butyl piperazine-1-carboxylate (10.94 g, 58.71 mmol, 5 eq) and 8-bromo-2-butyl-1-[4-[tert-butyl(dimethyl)silyl]oxybutyl]-N-[(2,4-dimethoxyphenyl)methyl]imidazo[4,5-c]quinolin-4-amine (7.7 g, 11.74 mmol, 1 eq) in dimethylformamide (DMF; 100 mL) was added 2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl (RuPhos, 547.95 mg, 1.17 mmol, 0.1 eq), Pd 2 (dba) 3  (537.64 mg, 587.12 μmol, 0.05 eq) and Cs 2 CO 3  (7.65 g, 23.48 mmol, 2 eq) in one portion at 20° C. under N 2 . The mixture was stirred at 130° C. for 2 hrs. The mixture was diluted with water (100 mL) and extracted with EtOAc (50 mL×3). The organic layer was washed with brine (50 mL), dried over Na 2 SO 4 , filtered and concentrated. The crude product was purified by chromatography on a silica gel eluted with petroleum ether:ethyl acetate (from 1/0 to 0/1). Compound tert-butyl 4-[2-butyl-1-[4-[tert-butyl(dimethyl)silyl]oxybutyl]-4-[(2,4-dimethoxyphenyl)methylamino]imidazo[4,5-c]quinolin-8-yl]piperazine-1-carboxylate (8 g, 9.98 mmol, 85.02% yield, 94.980% purity) was obtained as a yellow oil.  1 H NMR (DMSO-d 6 , 400 MHz) δ 7.49-7.56 (m, 1H), 7.26-7.32 (m, 1H), 7.19-7.23 (m, 1H), 7.15-7.19 (m, 1H), 6.54-6.58 (m, 1H), 6.46-6.53 (m, 1H), 6.37-6.42 (m, 1H), 4.65 (s, 2H), 4.47-4.53 (m, 2H), 3.81-3.85 (m, 3H), 3.68-3.73 (m, 3H), 3.58-3.65 (m, 2H), 3.47-3.54 (m, 4H), 3.10-3.16 (m, 4H), 2.87-2.93 (m, 2H), 1.72-1.95 (m, 6H), 1.60-1.66 (m, 2H), 1.42-1.44 (m, 9H), 0.93-0.97 (m, 3H), 0.78 (s, 9H), 0.00 (s, 6H). 
     Example 11: Synthesis of Compound 12 
     
       
         
         
             
             
         
       
     
     To a mixture of tert-butyl 4-[2-butyl-1-[4-[tert-butyl(dimethyl)silyl]oxybutyl]-4-[(2,4-dimethoxyphenyl)methylamino]imidazo[4,5-c]quinolin-8-yl]piperazine-1-carboxylate (8 g, 10.51 mmol, 1 eq) in dichloromethane (DCM; 100 mL) was added TFA (trifluoroacetic acid; 41.95 g, 367.90 mmol, 27.24 mL, 35 eq) in one portion at 20° C. The mixture was stirred at 50° C. for 12 hrs. The mixture was concentrated. Then the residue was diluted with MeOH (50 mL) and the mixture was filtered, the filtrate was concentrated to give the crude product 4-(4-amino-2-butyl-8-piperazin-1-yl-imidazo[4,5-c]quinolin-1-yl)butan-1-ol (8.1 g, crude, TFA) as a yellow solid for next step. 
     Example 12: Synthesis of Compound 13 
     
       
         
         
             
             
         
       
     
     To a solution of oxalyl chloride (127 mg, 86 μL, 1 mmol, 2 eq) in DCM (1 mL) at 80° C. was added dropwise a solution of DMSO (156 mg, 142 μL, 2 mmol, 4 eq) in DCM (1 mL). The mixture was stirred for 15 min at 80° C. To this mixture was added a solution of hydroxyl-PEG24-t-butyl ester (602 mg, 0.5 mmol, 1 eq) in DCM (1 mL). After stirring for 15 min, Et 3 N (303 mg, 418 μL) was added and the mixture was stirred at 80° C. for 15 min then removed from the cold bath and allowed to warm to 20° C. over 30 min. To a suspension of 4-(4-amino-2-butyl-8-piperazin-1-yl-imidazo[4,5-c]quinolin-1-yl)butan-1-ol TFA salt and sodium triacetoxyborohydride (212 mg, 1 mmol, 2 eq) in DMF (3 mL) was added the previous mixture slowly at 20° C. The combined mixture was stirred at 20° C. for 45 min. Solvent was removed under reduced pressure and to the remaining was added 3 mL of 10% Na 2 CO 3  and stirred vigorously for 15 min. Water (20 mL) was added and the crude product was extracted into DCM (25 mL). The organic layer was washed with brine, dried (Na 2 SO 4 ), filtered and concentrated. The crude material was purified by flash chromatography using a gradient elution of 2-15% MeOH/DCM+1% Et 3 N to yield tert-butyl 1-(4-(4-amino-2-butyl-1-(4-hydroxybutyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oate (490 mg, 0.31 mmol, 62%) as a golden syrup. LC/MS [M+H] 158.98 (calculated); LC/MS [M+H] 1582.27 (observed). 
     Example 13: Synthesis of Compound 14 
     
       
         
         
             
             
         
       
     
     To a solution of triphenylphosphine (0.236 mg, 0.9 mmol, 3 eq) in DCM (5 mL) was added diisopropyl azodicarboxylate (182 mg, 177 μL, 0.9 mmol, 3 eq) dropwise. After 5 min phthalimide (74 mg, 0.5 mmol, 1.7 eq) was added. The mixture was stirred for 5 minutes then a solution of tert-butyl 1-(4-(4-amino-2-butyl-1-(4-hydroxybutyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oate (475 mg, 0.3 mmol, 1 eq) in DCM (1 mL) was added to the mixture. After 30 min, the solvent was removed under reduced pressure and the crude material was purified by flash chromatography using a gradient elution of 2-10% MeOH/DCM+1% Et 3 N to yield tert-butyl 1-(4-(4-amino-2-butyl-1-(4-(1,3-dioxoisoindolin-2-yl)butyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oate (399 mg, 0.23 mmol, 77%) as a golden syrup. LC/MS [M+H] 1711.01 (calculated); LC/MS [M+H] 1711.25 (observed). 
     Example 14: Synthesis of Compound 15 
     
       
         
         
             
             
         
       
     
     A solution of tert-butyl 1-(4-(4-amino-2-butyl-1-(4-(1,3-dioxoisoindolin-2-yl)butyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oate (390 mg, 0.23 mmol, 1 eq) was combined with 1-butyl amine (3 mL) and heated at 80° C. in a capped vial in a heating block for 6 h. The solvent was removed and the crude material was purified by flash chromatography using a gradient elution of 2-15% MeOH/DCM+1% Et 3 N to yield tert-butyl 1-(4-(4-amino-1-(4-aminobutyl)-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oate (330 mg, 21 mmol, 92%) as a yellow syrup. LC/MS [M+H] 1581.00 (calculated); LC/MS [M+H] 1581.43 (observed). 
     Example 15: Synthesis of Compound 16 
     
       
         
         
             
             
         
       
     
     A mixture of yield tert-butyl 1-(4-(4-amino-1-(4-aminobutyl)-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oate (323 mg, 0.2 mmol, 1 eq) and 3-cyanophenylisocycanate were dissolved in DMF (3 mL) and heated at 80° C. in a capped vial in a heating block for 16 h. The solvent was removed and the crude material was purified by flash chromatography using a gradient elution of 2-10% MeOH/DCM+1% Et 3 N to yield tert-butyl 1-(4-(4-amino-2-butyl-1-(4-(3-(3-cyanophenyl)ureido)butyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oate (240 mg, 0.14 mmol, 70%) as a brownish solid. LC/MS [M+H] 1725.03 (calculated); LC/MS [M+H] 1725.30 (observed). 
     Example 16: Synthesis of Compound 17 
     
       
         
         
             
             
         
       
     
     Tert-butyl 1-(4-(4-amino-2-butyl-1-(4-(3-(3-cyanophenyl)ureido)butyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oate (225 mg, 0.14 mmol) was dissolved in a 1:1 mixture of dioxane and 3 N HCl (5 mL) then heated to 60° C. for 90 min. The solvent was removed and the residue was azeotroped four times with acetonitrile (5 mL). The resulting 1-(4-(4-amino-2-butyl-1-(4-(3-(3-cyanophenyl)ureido)butyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oic acid HCl salt (220 mg, 0.13 mmol, 95%) was used without further purification. 
     Example 17: Synthesis of Compound 18 
     
       
         
         
             
             
         
       
     
     To 1-(4-(4-amino-2-butyl-1-(4-(3-(3-cyanophenyl)ureido)butyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oic acid HCl salt (220 mg, 0.13 mmol, 1 eq) was added a mixture of 2,3,5,6-tetrafluorophenol (66 mg, 0.4 mmol, 3 eq) and diisopropylcarbodiimide (51 mg, 62 μL, 0.4 mmol, 3 eq) dissolved in acetonitrile (3 mL) and the mixture was stirred at 20° C. for 16 h. The mixture was diluted with water (12 mL) and purified by reverse phase chromatography using a gradient eluent of 30-80% acetonitrile/water+0.1% TFA over 10 min. The pooled fractions were concentrated under reduced pressure and the glassy film was azeotroped with acetonitrile four times (20 mL) to yield 2,3,5,6-tetrafluorophenyl 1-(4-(4-amino-2-butyl-1-(4-(3-(3-cyanophenyl)ureido)butyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oate (86 mg, 0.04 mmol, 36%) as a glassy film. LC/MS [M+H] 1816.96 (calculated); LC/MS [M+H] 1817.46 (observed). 
     Example 18: Synthesis of Compound 19 
     
       
         
         
             
             
         
       
     
     To a mixture of 6-bromo-2,4-dichloro-3-nitro-quinoline (5.5 g, 17.08 mmol, 1 eq) and tert-butyl N-(4-aminobutyl)carbamate (3.54 g, 18.79 mmol, 1.1 eq) in THE (20 mL) was added Et 3 N (2.59 g, 25.63 mmol, 3.57 mL, 1.5 eq) slowly in one portion at 0° C. The mixture was stirred at 0° C. for 2 h. TLC showed the reaction was finished and a new spot was detected. The mixture was diluted with ice-water and extracted with EtOAc (50 ml×3). The organic layer was washed with brine (40 ml), dried over Na 2 SO 4 , filtered and concentrated. The crude product was purified by re-crystallization from petroleum ether (200 mL) at 25° C. to give the tert-butyl N-[4-[(6-bromo-2-chloro-3-nitro-4-quinolyl)amino]butyl]carbamate (6.5 g, 13.7 mmol, 80.3% yield) as a yellow solid.  1 H NMR (DMSO-d 6 , 400 MHz) δ=8.80 (d, J=2.0 Hz, 1H), 7.95 (dd, J=2.0, 8.8 Hz, 1H), 7.74 (d, J=8.8 Hz, 1H), 6.78 (br t, J=5.2 Hz, 1H), 3.10 (br t, J=7.2 Hz, 2H), 2.93-2.85 (m, 2H), 1.61 (quin, J=7.4 Hz, 2H), 1.44-1.38 (m, 2H), 1.35 (s, 9H). 
     Example 19: Synthesis of Compound 20 
     
       
         
         
             
             
         
       
     
     To a solution of tert-butyl N-[4-[(6-bromo-2-chloro-3-nitro-4-quinolyl)amino]butyl]carbamate (5.5 g, 11.61 mmol, 1 eq) in EtOAc (20 mL) was added platinum on carbon (2.44 g, 5% purity). The suspension was degassed under vacuum and purged with H 2  several times. The mixture was stirred under H 2  (50 psi) at 25° C. for 2 h. LCMS showed the reaction was finished. The mixture was filtered and concentrated. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether/ethyl acetate=4/1, 1/1) to yield tert-butyl N-[4-[(3-amino-6-bromo-2-chloro-4-quinolyl) amino]butyl]carbamate (3.4 g, 7.6 mmol, 66% yield) as a yellow solid.  1 H NMR (400 MHz, DMSO-d 6 ) δ=8.23 (d, J=2.0 Hz, 1H), 7.62 (d, J=8.8 Hz, 1H), 7.55-7.46 (m, 1H), 6.75 (m, 1H), 5.28 (t, J=6.4 Hz, 1H), 3.13-3.19 (m, 2H), 2.92-2.78 (m, 2H), 1.45-1.50 (m, 2H), 1.43-1.37 (m, 2H), 1.34 (s, 9H). 
     Example 20: Synthesis of Compound 21 
     
       
         
         
             
             
         
       
     
     To a mixture of tert-butyl N-[4-[(3-amino-6-bromo-2-chloro-4-quinolyl) amino]butyl]carbamate (1.7 g, 3.83 mmol, 1 eq) and pentanoyl chloride (924 mg, 7.6 mmol, 930 μL, 2 eq) in THE (30 ml) was added Et 3 N (582 mg, 5.8 mmol, 800 μL, 1.5 eq) in one portion at 0° C. Then the mixture was stirred at 25° C. for 2 h. LCMS showed the reaction was finished. The mixture was diluted with water (30 ml) and extracted with EtOAc (30 mL×3). The organic layer was washed with brine (40 ml), dried over Na 2 SO 4 , filtered and concentrated. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether/ethyl acetate=4/1, 1/1) to yield tert-butyl N-[4-[[6-bromo-2-chloro-3-(pentanoylamino)-4-quinolyl]amino]butyl]carbamate (2 g, 3.79 mmol, 99% yield) as a yellow solid.  1 H NMR (DMSO-d 6 , 400 MHz) δ=8.56 (d, J=2.0 Hz, 1H), 7.78 (dd, J=2.0, 8.8 Hz, 1H), 7.63 (d, J=8.8 Hz, 1H), 6.90-6.85 (m, 1H), 6.80-6.78 (m, 1H), 3.43-3.38 (m, 2H), 2.94-2.89 (m, 2H), 2.39-2.29 (m, 2H), 1.64-1.50 (m, 4H), 1.36 (s, 9H), 0.94-0.88 (m, 3H) 
     Example 21: Synthesis of Compound 22 
     
       
         
         
             
             
         
       
     
     To a mixture of tert-butyl N-[4-(8-bromo-2-butyl-4-chloro-imidazo[4,5-c]quinolin-1-yl)butyl]carbamate (0.8 g, 1.6 mmol, 1 eq) and (2,4-dimethoxyphenyl) methanamine (2.62 g, 15.7 mmol, 2.36 mL, 10 eq) was stirred at 120° C. for 2 hours. The mixture was added 2M HCl adjust to pH˜4 and extracted with ethyl acetate (50 mL×3). The combined organic phase was washed with brine (50 mL), dried with anhydrous Na 2 SO 4 , filtered and concentrated in vacuum. The residue was purified by column chromatography (SiO 2 , petroleum ether/ethyl acetate=20/1 to 0:1). Compound tert-butyl N-[4-[8-bromo-2-butyl-4-[(2,4-dimethoxyphenyl) methylamino] imidazo[4,5-c]quinolin-1-yl]butyl]carbamate (0.97 g, 1.51 mmol, 97% yield) was obtained as a yellow solid.  1 H NMR (MeOD, 400 MHz) δ 8.19 (s, 1H), 7.91-7.70 (m, 2H), 7.28 (d, J=8.4 Hz, 1H), 6.59 (s, 1H), 6.51 (d, J=8.4 Hz, 1H), 4.84-4.78 (m, 2H), 4.56 (t, J=7.6 Hz, 2H), 3.83 (s, 3H), 3.78 (s, 3H), 3.17-3.06 (m, 2H), 2.98 (t, J=7.8 Hz, 2H), 1.98-1.86 (m, 4H), 1.70-1.60 (m, 2H), 1.54-1.49 (m, 2H), 1.36 (s, 9H), 1.01 (t, J=7.6 Hz, 2H). 
     Example 22: Synthesis of Compound 23 
     
       
         
         
             
             
         
       
     
     A solution of tert-butyl N-[4-[8-bromo-2-butyl-4-[(2,4-dimethoxyphenyl) methylamino]imidazo[4,5-c]quinolin-1-yl]butyl]carbamate (0.47 g, 733.68 μmol, 1 eq) in HCl/EtOAc (4 M, 50 mL) was stirred at 25° C. for 2 hours. The mixture was concentrated under reduced pressure at 45° C. Compound 1-(4-aminobutyl)-8-bromo-2-butyl-N-[(2,4-dimethoxyphenyl)methyl]imidazo[4,5-c]quinolin-4-amine (0.5 g, crude, HCl) was obtained as a yellow solid. LCMS (ESI): mass calcd. for C 27 H 34 N 5 O 2 Br 539.2/541.2, m/z found 540.3/542.3 [M+H] +   
     Example 23: Synthesis of Compound 24 
     
       
         
         
             
             
         
       
     
     A mixture of 1-(4-aminobutyl)-8-bromo-2-butyl-N-[(2,4-dimethoxyphenyl) methyl]imidazo[4,5-c]quinolin-4-amine (0.4 g, 740.06 μmol, 1 eq) and triethylamine (TEA; 149.77 mg, 1.48 mmol, 206.02 μL, 2 eq) in MeOH (20 mL) was stirred at 25° C. for 30 min then formaldehyde (132.13 mg, 1.63 mmol, 121.22 μL, 2.2 eq) and AcOH (44.44 mg, 740.06 μmol, 42.32 μL, 1 eq) was added to the mixture stirred 30 min at 25° C. NaBH 3 CN (186.02 mg, 2.96 mmol, 4 eq) was added to the mixture at 0° C. and stirred at 25° C. for 1.5 hours. The mixture was concentrated under reduced pressure at 45° C. The residue was added water (50 mL) and extracted with ethyl acetate (50 mL×3). The combined organic phase was washed with brine (50 mL), dried with anhydrous Na 2 SO 4 , filtered and concentrated in vacuum. Compound 8-bromo-2-butyl-N-[(2,4-dimethoxyphenyl)methyl]-1-[4-(dimethylamino)butyl]imidazo[4,5-c]quinolin-4-amine (0.5 g, crude) was obtained as a yellow solid.  1 H NMR (MeOD, 400 MHz) δ 8.06 (m, 1H), 7.71 (d, J=9.0 Hz, 1H), 7.54 (d, J=10.4 Hz, 1H), 7.29 (d, J=8.4 Hz, 1H), 6.56 (d, J=2.4 Hz, 1H), 6.46 (dd, J=2.4, 8.4 Hz, 1H), 4.74 (s, 2H), 4.48-4.46 (m, 2H), 3.86 (s, 3H), 3.77 (s, 3H), 2.96-2.92 (m, 2H), 2.43-2.40 (m, 2H), 2.26 (s, 6H), 1.94-1.81 (m, 4H), 1.70-1.68 (m, 2H), 1.56-1.47 (m, 2H), 1.24 (t, J=7.2 Hz, 3H), 1.01 (t, J=7.2 Hz, 3H). 
     Example 24: Synthesis of Compound 25 
     
       
         
         
             
             
         
       
     
     To a mixture of 8-bromo-2-butyl-N-[(2,4-dimethoxyphenyl)methyl]-1-[4-(dimethylamino)butyl]imidazo[4,5-c]quinolin-4-amine (0.48 g, 844.26 μmol, 1 eq) and tert-butyl piperazine-1-carboxylate (786.22 mg, 4.22 mmol, 5 eq) in DMF (50 mL) were added Cs 2 CO 3  (550.15 mg, 1.69 mmol, 2 eq), RuPhos (39.40 mg, 84.43 μmol, 0.1 eq) and Pd 2 (dba) 3  (38.66 mg, 42.21 μmol, 0.05 eq) in one portion at 25° C. under N 2 . The mixture was stirred at 120° C. for 2 hours. The mixture was added H 2 O (150 mL) and extracted with ethyl acetate (50 mL×3). The combined organic phase was washed with brine (50 mL×2), dried with anhydrous Na 2 SO 4 , filtered and concentrated in vacuum. Compound tert-butyl 4-[2-butyl-4-[(2,4-dimethoxyphenyl)methylamino]-1-[4-(dimethylamino)butyl]imidazo[4,5-c]quinolin-8-yl]piperazine-1-carboxylate (0.5 g, crude) was obtained as a yellow solid. LCMS (ESI): mass calcd. for C 38 H 55 N 7 O 4  673.4, m/z found 674.4 [M+H] + . 
     Example 25: Synthesis of Compound 26 
     
       
         
         
             
             
         
       
     
     To a solution of tert-butyl 4-[2-butyl-4-[(2,4-dimethoxyphenyl) methylamino]-1-[4-(dimethylamino)butyl]imidazo[4,5-c]quinolin-8-yl]piperazine-1-carboxylate (0.5 g, 741.97 μmol, 1 eq) in DCM (20 mL) was added TFA (2.57 g, 22.51 mmol, 1.67 mL, 30.34 eq) in one portion at 25° C. The mixture was stirred at 40° C. for 12 hours. The mixture was concentrated in reduced pressure at 45° C. The residue was purified by prep-HPLC (column: LUNA™ C18 100×30 5 u (Phenomenex, Inc., Torrance, Calif.); mobile phase: [water (0.1% TFA)-ACN]; B %: 5%-30%, 10 min). Compound 2-butyl-1-[4-(dimethylamino)butyl]-8-piperazin-1-yl-imidazo[4,5-c]quinolin-4-amine (0.130 g, 306.90 μmol, 41.36% yield) was obtained as a yellow oil. The compound was used in the subsequent step without further purification. 
     Example 26: Synthesis of Compound 27 
     
       
         
         
             
             
         
       
     
     To a solution of 2-butyl-1-[4-(dimethylamino) butyl]-8-piperazin-1-yl-imidazo [4,5-c]quinolin-4-amine (0.13 g, 199.50 μmol, 1 eq, 2TFA) in MeOH (20 mL) was added TEA (40.37 mg, 398.99 μmol, 55.54 μL, 2 eq) at 25° C. and the mixture was stirred for 0.5 hour. Then tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-oxoethoxy) ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate (116.64 mg, 199.50 μmol, 1 eq), AcOH (11.98 mg, 199.50 μmol, 11.41 μL, 1 eq) and NaBH 3 CN (25.07 mg, 398.99 μmol, 2 eq) were added at 25° C. and stirred for 11.5 hours. The mixture was added H 2 O (2 mL) and concentrated in reduced pressure at 50° C. Compound tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-[4-amino-2-butyl-1-[4-(dimethylamino)butyl]imidazo[4,5-c]quinolin-8-yl]piperazin-1-yl]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate (0.2 g, crude) was obtained as a yellow oil. LC/MS [M+H] 992.66 (calculated); LC/MS [M+H] 993.08 (observed). 
     Example 27: Synthesis of Compound 28 
     
       
         
         
             
             
         
       
     
     To a solution of tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-[4-amino-2-butyl-1-[4-(dimethylamino)butyl]imidazo[4,5-c]quinolin-8-yl]piperazin-1-yl]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate (0.2 g, 201.55 μmol, 1 eq) in H 2 O (10 mL) was added TFA (22.98 mg, 201.55 μmol, 14.92 μL, 1 eq) at 60° C. and stirred for 12 hours. The mixture was concentrated under reduced pressure at 50° C. The residue was purified by prep-HPLC (column: Nano-micro KROMASIL™ (Sigma-Aldrich, St. Louis, Mo.) C18 100×30 mm Sum; mobile phase: [water (0.1% TFA)-ACN]; B %: 15%-35%, 10 min). Compound 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-[4-amino-2-butyl-1-[4-(dimethylamino)butyl]imidazo[4,5-c]quinolin-8-yl]piperazin-1-yl]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (0.0887 g, 94.75 μmol, 47.01% yield, 100% purity) was obtained as a yellow oil.  1 H NMR (MeOD, 400 MHz) δ 7.76 (d, J=9.2 Hz, 1H), 7.56 (d, J=9.2 Hz, 1H), 7.51 (s, 1H), 4.73 (t, J=7.2 Hz 2H), 4.09-3.78 (m, 6H), 3.76-3.45 (m, 44H), 3.25-3.15 (m, 2H), 3.04 (t, J=7.6 Hz, 2H), 2.88 (s, 6H), 2.52 (t, J=6.0 Hz, 2H), 2.12-1.89 (m, 6H), 1.57-1.53 (m, 2H), 1.04 (t, J=7.2 Hz, 3H). LCMS (ESI): mass calcd. for C 41 H 81 N 7 O 12 =935.6, m/z found 936.2 [M+H] + . 
     Example 28: Synthesis of Compound 29 
     
       
         
         
             
             
         
       
     
     1-(4-(4-amino-2-butyl-1-(4-(dimethylamino)butyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oic acid HCl was converted into 2,3,5,6-tetrafluorophenyl 1-(4-(4-amino-2-butyl-1-(4-(dimethylamino)butyl)-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate in a 46% yield using the procedure described in Example 17. LC/MS [M+H] 1084.60 (calculated); LC/MS [M+H] 1084.86 (observed). 
     Example 29: Synthesis of Compound 30 
     
       
         
         
             
             
         
       
     
     To a mixture of tert-butyl N-[4-[8-bromo-2-butyl-4-[(2,4-dimethoxyphenyl) methylamino]imidazo[4,5-c]quinolin-1-yl]butyl]carbamate (0.5 g, 780.51 μmol, 1 eq) and 1-methylpiperazine (234.53 mg, 2.34 mmol, 259.72 μL, 3 eq) in DMF (20 mL) were added Cs 2 CO 3  (508.61 mg, 1.56 mmol, 2 eq), RuPhos (36.42 mg, 78.05 μmol, 0.1 eq) and Pd 2 (dba) 3  (35.74 mg, 39.03 μmol, 0.05 eq) in one portion at 25° C. under N 2 . The mixture was stirred at 120° C. for 2 hours. To the mixture was added H 2 O (80 mL) and extracted with ethyl acetate (50 mL×3). The combined organic phase was washed with brine (30 mL×2), dried with anhydrous Na 2 SO 4 , filtered and concentrated in vacuum. The residue was purified by column chromatography (SiO 2 , ethyl acetate:MeOH=10:1). Compound tert-butyl N-[4-[2-butyl-4-[(2,4-dimethoxyphenyl)methylamino]-8-(4-methylpiperazin-1-yl)imidazo[4,5-c]quinolin-1-yl]butyl]carbamate (0.27 g, 409.18 μmol, 52.42% yield) was obtained as a yellow oil.  1 H NMR (MeOD, 400 MHz) δ 7.77 (d, J=9.2 Hz, 1H), 7.37 (s, 1H), 7.34-7.27 (m, 2H), 6.56 (s, 1H), 6.46 (d, J=8.4 Hz, 1H), 4.72 (s, 2H), 4.56-4.45 (m, 2H), 3.85 (s, 3H), 3.80 (s, 3H), 3.31-3.28 (m, 2H), 2.93 (t, J=7.6 Hz, 1H), 2.70-2.68 (m, 4H), 2.39 (s, 3H), 1.93-1.91 (m, 2H), 1.87-1.77 (m, 2H), 1.64-1.62 (m, 2H), 1.55-1.46 (m, 2H), 1.35 (s, 9H), 1.24 (t, J=7.6 Hz, 2H), 1.01 (t, J=7.2 Hz, 3H). 
     Example 30: Synthesis of Compound 31 
     
       
         
         
             
             
         
       
     
     To a solution of tert-butyl N-[4-[2-butyl-4-[(2,4-dimethoxyphenyl)methylamino]-8-(4-methylpiperazin-1-yl)imidazo[4,5-c]quinolin-1-yl]butyl]carbamate (0.22 g, 333.40 μmol, 1 eq) in THF (20 mL) was added LiAlH 4  (63.27 mg, 1.67 mmol, 5 eq) in portions at 25° C. under N 2 . The mixture was stirred at 60° C. for 3 hours. The mixture was added saturated aqueous Na 2 SO 4  (2 mL) at 0° C. and dried with anhydrous Na 2 SO 4 , filtered and concentrated in vacuum. Compound 2-butyl-N-[(2,4-dimethoxyphenyl) methyl]-1-[4-(methylamino)butyl]-8-(4-methylpiperazin-1-yl)imidazo[4,5-c]quinolin-4-amine (0.2 g, crude) was obtained as a yellow oil.  1 H NMR (CDCl 3 , 400 MHz) δ 7.82 (d, J=9.2 Hz, 1H), 7.41 (d, J=8.2 Hz, 1H), 7.32 (d, J=2.4 Hz, 1H), 7.24 (d, J=2.4 Hz, 1H), 6.47 (d, J=2.2 Hz, 1H), 6.42 (dd, J=2.4, 8.2 Hz, 1H), 4.83 (s, 2H), 4.43 (t, J=3.6 Hz, 2H), 4.16-4.06 (m, 2H), 3.85 (s, 3H), 3.79 (s, 3H), 3.73-3.63 (m, 2H), 3.31-3.23 (m, 4H), 2.87 (t, J=8.0 Hz, 2H), 2.71-2.64 (m, 4H), 2.41 (d, J=7.2 Hz, 4H), 2.01-1.87 (m, 2H), 1.90-1.80 (m, 2H), 1.76-1.63 (m, 4H), 0.99 (t, J=7.6 Hz, 3H). 
     Example 31: Synthesis of Compound 32 
     
       
         
         
             
             
         
       
     
     To a solution of 2-butyl-N-[(2,4-dimethoxyphenyl) methyl]-1-[4-(methylamino) butyl]-8-(4-methylpiperazin-1-yl)imidazo[4,5-c]quinolin-4-amine (0.2 g, 348.57 μmol, 1 eq) in DCM (20 mL) was added TFA (2.80 g, 24.56 mmol, 1.82 mL, 70.45 eq) in one portion at 25° C. The mixture was stirred at 40° C. for 12 hours. The mixture was concentrated in reduced pressure at 45° C. The residue was purified by prep-HPLC (column: LUNA™ C18 100×30 5 u (Phenomenex, Inc.); mobile phase: [water (0.10% TFA)-ACN]; B %: 5%-25%, 10 min). Compound 2-butyl-1-[4-(methylamino) butyl]-8-(4-methylpiperazin-1-yl) imidazo[4,5-c]quinolin-4-amine (0.2 g, crude) was obtained as a yellow oil. LC/MS [M+H]424.32 (calculated); LC/MS [M+H] 424.43 (observed). 
     Example 32: Synthesis of Compound 33 
     
       
         
         
             
             
         
       
     
     To a solution of 2-butyl-1-[4-(methylamino)butyl]-8-(4-methylpiperazin-1-yl) imidazo[4,5-c]quinolin-4-amine (0.1 g, 153.46 μmol, 1 eq, 2TFA) in MeOH (20 mL) was added TEA (31.06 mg, 306.92 μmol, 42.72 μL, 2 eq) at 25° C. and stirred for 0.5 hour. Then tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-oxoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate (89.73 mg, 153.46 μmol, 1 eq), AcOH (9.22 mg, 153.46 μmol, 8.78 μL, 1 eq) and NaBH 3 CN (19.29 mg, 306.92 μmol, 2 eq) were added at 25° C. and stirred for 11.5 hours. The mixture was added to H 2 O (2 mL) and concentrated in reduced pressure at 50° C. Compound tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-[4-amino-2-butyl-8-(4-methylpiperazin-1-yl)imidazo[4,5-c]quinolin-1-yl]butyl-methyl-amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate (0.2 g, crude) was obtained as a yellow oil, which was used without purification in the next step. 
     Example 33: Synthesis of Compound 34 
     
       
         
         
             
             
         
       
     
     To a solution of tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-[4-amino-2-butyl-8-(4-methylpiperazin-1-yl)imidazo[4,5-c]quinolin-1-yl]butyl-methyl-amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate (0.15 g, 151.17 μmol, 1 eq) in H 2 O (10 mL) was added TFA (1.54 g, 13.51 mmol, 1 mL, 89.35 eq) at 25° C. and stirred at 60° C. for 12 hours. The mixture was concentrated in reduced pressure at 50° C. The residue was purified by prep-HPLC (column: Nano-micro KROMASIL™ (Sigma-Aldrich) C18 100×30 mm Sum; mobile phase: [water (0.1% TFA)-ACN]; B %: 15%-35%, 10 min). Compound 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-[4-amino- 2-butyl-8-(4-methylpiperazin-1-yl)imidazo[4,5-c]quinolin-1-yl]butyl-methyl-amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (0.0444 g, 47.43 μmol, 31.37% yield, 100% purity) was obtained as a yellow oil.  1 H NMR (MeOD, 400 MHz) δ 7.77 (d, J=9.2 Hz, 1H), 7.60-7.50 (m, 2H), 4.75 (t, J=7.0 Hz, 2H), 4.00 (d, J=10.4 Hz, 2H), 3.82 (t, J=4.8 Hz, 2H), 3.74-3.70 (m, 4H), 3.65-3.49 (m, 40H), 3.05 (t, J=7.8 Hz, 2H), 3.02 (s, 3H), 2.92 (s, 3H), 2.53 (t, J=6.2 Hz, 2H), 2.09-1.87 (m, 6H), 1.61-1.52 (m, 2H), 1.05 (t, J=7.6 Hz, 3H). LCMS (ESI): mass calcd. for C 47 H 81 N 7 O 12  935.6, m/z found 936.2 [M+H] + . 
     Example 34: Synthesis of Compound 35 
     
       
         
         
             
             
         
       
     
     38-(4-amino-2-butyl-8-(4-methylpiperazin-1-yl)-1H-imidazo[4,5-c]quinolin-1-yl)-34-methyl-4,7,10,13,16,19,22,25,28,31-decaoxa-34-azaoctatriacontanoic acid HCl was converted into 2,3,5,6-tetrafluorophenyl 38-(4-amino-2-butyl-8-(4-methylpiperazin-1-yl)-1H-imidazo[4,5-c]quinolin-1-yl)-34-methyl-4,7,10,13,16,19,22,25,28,31-decaoxa-34-azaoctatriacontanoate TFA in a 52% yield using the procedure described in Example 17. LC/MS [M+Na] 1106.58 (calculated); LC/MS [M+Na] 1107.00 (observed). 
     Example 35: Synthesis of Compound 36 
     
       
         
         
             
             
         
       
     
     To a solution of 6-bromo-2,4-dichloro-3-nitroquinoline (5.6 g, 17.4 mmol, 1 eq.) and solid K2CO 3  (3.6 g, 26 mmol, 1.5 eq.) in DMF (100 mL) at room temperature was added neat 2,4-dimethoxybenzylamine (3.5 g, 20.1 mmol, 1.2 eq.). The mixture was stirred for 15 minutes, water (300 mL) was added and the mixture was stirred for 5 additional minutes. The resultant solid was filtered and then dissolved in ethyl acetate (100 mL). The solution was washed with water (100 mL), brine (100 mL), separated, dried (Na 2 SO 4 ), then filtered and concentrated in vacuo. The brown solid was triturated with 1:1 hexanes/diethyl ether (150 mL) and filtered to obtain 6-bromo-2-chloro-4-(2,4-dimethoxybenzyl)amino-3-nitroquinoline (6.9 g, 15.3 mmol, 88%) as a yellow solid. The compound was used without further purification. 
     Example 36: Synthesis of Compound 37 
     
       
         
         
             
             
         
       
     
     To 6-bromo-2-chloro-4-(2,4-dimethoxybenzyl)amino-3-nitroquinoline (6.9 g, 15.3 mmol, 88%) in methanol (200 mL) at 0° C. was added NiCl 2 .6H 2 O (0.36 g, 1.5 mmol, 0.1 eq). Sodium borohydride (pellets, 1.42 g, 38 mmol, 2.5 eq.) was added and reaction was stirred for 1 h at 0° C. then warmed to room temperature and allowed to stir for another 15 minutes. Glacial acetic acid (5 mL) was added until a pH of ˜5 was obtained. The solvent was evaporated in vacuo and the crude solid was redissolved in ethyl acetate (150 mL) then filtered through a bed of diatomaceous earth to remove a black insoluble material. The ethyl acetate was removed in vacuo. The dark brown solid was triturated with ether (75 mL) then filtered to obtain 3-amino-6-bromo-2-chloro-4-(2,4-dimethoxybenzyl)aminoquinoline (5.81 g, 13.7 mmol, 90%) as a tan solid. The compound was used without further purification. 
     Example 37: Synthesis of Compound 38 
     
       
         
         
             
             
         
       
     
     To a solution of 3-amino-6-bromo-2-chloro-4-(2,4-dimethoxybenzyl)aminoquinoline (5.75 g, 13.6 mmol, 1 eq.) in dichloromethane (100 mL) containing triethylamine (2.1 g, 2.8 mL, 20 mmol, 1.5 eq.) stirring at room temperature was added neat valeroyl chloride (2.0 mL, 2.0 g, 16 mmol, 1.2 eq). The mixture was washed with water (150 mL), brine (150 mL), separated, dried (Na 2 SO 4 ), filtered, and concentrated. The solid was triturated with ether, filtered and dried under vacuum. N-(6-bromo-2-chloro-4-((2,4-dimethoxybenzyl)amino)quinolin-3-yl)pentanamide was obtained as a brown solid (5.8 g, 11.4 mmol, 84%). The compound was used without further purification. 
     Example 38: Synthesis of Compound 39 
     
       
         
         
             
             
         
       
     
     In a 100 mL beaker a mixture of N-(6-bromo-2-chloro-4-((2,4-dimethoxybenzyl)amino)quinolin-3-yl)pentanamide (5.8 g, 11.4 mmol, 1 eq.) and 2-chlorobenzoic (0.90 g, 5.7 mmol. 0.5 eq.) was boiled in 50 mL toluene for 2 hours. Toluene was added to 50 mL each time the volume reached 25 mL. 2,4-dimethoxybenzylamine (9.5 g, 57 mmol, 5 eq.) was added and the reaction was maintained at 120° C. for 2 hours. The reaction was cooled to room temperature and water (80 mL) then acetic acid (3.5 mL) was added. The supernatant was decanted and the crude product was washed with water (80 mL). The wet solid was triturated with methanol (100 mL) to provide 8-bromo-2-butyl-N,1-bis(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinolin-4-amine (4.80 g, 7.7 mmol, 68%) as an off-white solid. The compound was used without further purification. 
     Example 39: Synthesis of Compound 40 
     
       
         
         
             
             
         
       
     
     A mixture of 8-bromo-2-butyl-N,1-bis(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinolin-4-amine (0.31 g, 0.5 mmol, 1 eq.) and tert-butyl piperazine-1-carboxylate (0.19 g, 1 mmol, 2 eq.) were combined in toluene (2 mL) then degassed with argon. Pd 2 dba 3  (45 mg, 0.05 mmol, 0.1 eq.), tri-tert-butylphosphine tetrafluoroborate (29 mg, 0.10 mmol, 0.2 eq) and sodium tert-butoxide (144 mg, 1.5 mmol, 3 eq) were added. The mixture was heated in a capped vial at 110° C. for 30 minutes. The mixture was cooled then partitioned between ethyl acetate (50 mL) and water (50 mL). The organic layer was washed with brine (50 mL), dried with sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified on silica gel (20 g) eluted with 50% ethyl acetate/hexanes to yield tert-butyl 4-(2-butyl-1-(2,4-dimethoxybenzyl)-4-((2,4-dimethoxybenzyl)amino)-1H-imidazo[4,5-c]quinolin-8-yl)piperazine-1-carboxylate (0.28 g, 0.39 mmol, 78%) as an off-white solid. LC/MS [M+H] 725.40 (calculated); LC/MS [M+H] 725.67 (observed). 
     Example 40: Synthesis of Compound 41 
     
       
         
         
             
             
         
       
     
     Tert-butyl 4-(2-butyl-1-(2,4-dimethoxybenzyl)-4-((2,4-dimethoxybenzyl)amino)-1H-imidazo[4,5-c]quinolin-8-yl)piperazine-1-carboxylate (0.28 g, 0.39 mmol, 1 eq.) was dissolved in TFA (3 mL) and heated to reflux for 5 min. The TFA was removed in vacuo and the crude product was dissolved in acetonitrile, filtered then concentrated to obtain the TFA salt of 2-butyl-8-(piperazin-1-yl)-1H-imidazo[4,5-c]quinolin-4-amine (0.16 g, 0.37 mmol, 95%) as an off-white solid. LC/MS [M+H] 325.21 (calculated); LC/MS [M+H] 325.51 (observed). 
     Example 41: Synthesis of Compound 42 
     
       
         
         
             
             
         
       
     
     2-butyl-8-(piperazin-1-yl)-1H-imidazo[4,5-c]quinolin-4-amine was converted into tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate in a 56% yield using the procedure described in Example 12. LC/MS [M+H] 893.55 (calculated); LC/MS [M+H] 893.79 (observed). 
     Example 42: Synthesis of Compound 43 
     
       
         
         
             
             
         
       
     
     Tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate was converted into 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oic acid in a 93% yield using the procedure described in Example 16. The compound was used without further purification. 
     Example 43: Synthesis of Compound 44 
     
       
         
         
             
             
         
       
     
     1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oic acid was converted into 2,3,5,6-tetrafluorophenyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate in a 54% yield using the procedure described in Example 17. LC/MS [M+H] 985.49 (calculated); LC/MS [M+H] 985.71 (observed). 
     Example 44: Synthesis of Compound 46 
     
       
         
         
             
             
         
       
     
     To a mixture of 6-bromo-1H-indole (5.00 g, 25.50 mmol, 1 eq) and pyridine (2.62 g, 33.16 mmol, 2.68 mL, 1.3 eq) in Et 2 O (80 mL) was added ethyl 2-chloro-2-oxo-acetate (4.18 g, 30.61 mmol, 3.43 mL, 1.2 eq) slowly at 0° C. under N 2 . The mixture was stirred at 0° C. for 2 hours. A yellow solid precipitated. The mixture was filtered and the cake was washed by H 2 O. The crude product was triturated with H 2 O at 20° C. for 20 min to provide ethyl 2-(6-bromo-1H-indol-3-yl)-2-oxo-acetate (5.4 g, 18.24 mmol, 71.50% yield) as a yellow solid.  1 H NMR (DMSO-d 6 , 400 MHz) δ 12.46 (s, 1H), 8.46 (d, J=3.6 Hz, 1H), 8.10 (d, J=8.8 Hz, 1H), 7.75 (s, 1H), 7.43 (d, J=8.8 Hz, 1H), 4.36 (q, J=7.2 Hz, 2H), 1.33 (t, J=7.2 Hz, 3H). 
     Example 45: Synthesis of Compound 47 
     
       
         
         
             
             
         
       
     
     To a mixture of ethyl 2-(6-bromo-1H-indol-3-yl)-2-oxo-acetate (5.4 g, 18.24 mmol, 1 eq) and butylhydrazine (3.41 g, 27.35 mmol, 1.5 eq, HCl) in EtOH (60 mL) was added AcOH (10.95 g, 182.36 mmol, 10.43 mL, 10 eq) at 25° C. under N 2 . The mixture was stirred at 90° C. for 16 hours. LCMS showed the reaction was completed. The mixture was concentrated in vacuum. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether/ethyl acetate=5/1, 1/2) to provide 7-bromo-2-butyl-pyrazolo[3,4-c] quinolin-4-ol (3 g, 9.37 mmol, 51.38% yield) as a brown solid.  1 H NMR (CDCl 3 , 400 MHz) δ 11.40 (s, 1H), 8.72 (s, 1H), 7.81 (d, J=8.4 Hz, 1H), 7.50 (s, 1H), 7.34 (dd, J=8.4, 2.0 Hz, 1H), 4.37 (t, J=6.8 Hz, 2H), 1.91-1.84 (m, 2H), 1.32-1.25 (m, 2H), 0.91 (t, J=7.2 Hz, 3H). 
     Example 46: Synthesis of Compound 48 
     
       
         
         
             
             
         
       
     
     To a mixture of 7-bromo-2-butyl-pyrazolo[3,4-c]quinolin-4-ol (2.8 g, 8.74 mmol, 1 eq) in POCl 3  (13.41 g, 87.45 mmol, 8.13 mL, 10 eq) was added PCl 5  (910.52 mg, 4.37 mmol, 0.5 eq) in one portion at 25° C. The mixture was stirred at 100° C. for 1 hour. LCMS showed the reaction was completed. The mixture was concentrated. The residue was poured into ice water (100 mL) and diluted with CH 2 C 2  (30 mL) and washed with saturated NaHCO 3 , dried over Na 2 SO 4 , filtered, and concentrated. 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, 3/1) to provide 7-bromo-2-butyl-4-chloro-pyrazolo[3,4-c]quinoline (2.6 g, 7.68 mmol, 87.80% yield) as a yellow oil.  1 H NMR (DMSO-d 6 , 400 MHz) δ 8.30 (s, 1H), 8.22 (d, J=2.0 Hz, 1H), 7.85 (d, J=8.4 Hz, 1H), 7.68 (dd, J=8.4, 2.0 Hz, 1H), 4.53 (t, J=7.2 Hz, 2H), 2.08-2.04 (m, 2H), 1.46-1.37 (m, 2H), 0.10 (t, J=7.2 Hz, 3H). 
     Example 47: Synthesis of Compound 49 
     
       
         
         
             
             
         
       
     
     To mixture 7-bromo-2-butyl-4-chloro-pyrazolo[3,4-c]quinoline (2.6 g, 7.68 mmol, 1 eq) in 2,4-dimethoxyphenyl)methanamine (6.42 g, 38.39 mmol, 5.78 mL, 5 eq) was stirred at 120° C. for 4 hours. LCMS showed the reaction was completed. The mixture was dissolved in EtOAc/H 2 O (10 mL/10 mL) and adjusted pH=3 with aq. HCl (4 M). The aqueous phase was filtered to give 7-bromo-2-butyl-N-[(2,4-dimethoxyphenyl)methyl]pyrazolo[3,4-c]quinolin-4-amine (2.9 g, 6.18 mmol, 80.47% yield) as a yellow solid which was used into the next step without further purification.  1 H NMR (CDCl 3 , 400 MHz) δ 9.03 (s, 1H), 8.34 (s, 1H), 8.04 (d, J=8.4 Hz, 1H), 7.64 (s, 1H), 7.20 (d, J=8.4 Hz, 1H), 6.61 (d, J=2.4 Hz, 1H), 6.51 (d, J=8.4 Hz, 1H), 4.89 (d, J=4.2 Hz, 2H), 4.49 (t, J=6.8 Hz, 2H), 3.75 (m, 6H), 1.96-1.89 (m, 2H), 1.35-1.27 (m, 2H), 0.91 (t, J=7.2 Hz, 3H). 
     Example 48: Synthesis of Compound 50 
     
       
         
         
             
             
         
       
     
     To a mixture of 7-bromo-2-butyl-N-[(2,4-dimethoxyphenyl)methyl]pyrazolo[3,4-c] quinolin-4-amine (0.45 g, 958.73 μmol, 1 eq) and tert-butyl piperazine-1-carboxylate (535.69 mg, 2.88 mmol, 3 eq) in DMF (10 mL) was added Pd 2 (dba) 3  (43.90 mg, 47.94 μmol, 0.05 eq), Cs 2 CO 3  (624.74 mg, 1.92 mmol, 2 eq) and RuPhos (44.74 mg, 95.87 μmol, 0.1 eq) in one portion at 25° C. under N 2 . The mixture was stirred at 140° C. for 2 hours. LCMS showed the reaction was completed. The mixture was cooled to 25° C. and poured into ice water (30 mL) and stirred for 1 min. The aqueous phase was extracted with ethyl acetate (10 mL×3). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na 2 SO 4 , filtered, and concentrated in vacuum. The residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=10/1, 1/1) to provide tert-butyl 4-[2-butyl-4-[(2,4-dimethoxyphenyl) methylamino]pyrazolo[3,4-c]quinolin-7-yl]piperazine-1-carboxylate (0.45 g, 783.00 μmol, 81.67% yield) as a yellow oil.  1 H NMR (CDCl 3 , 400 MHz) δ 7.95 (s, 1H), 7.67 (d, J=8.8 Hz, 1H), 7.40 (d, J=8.0 Hz, 1H), 7.28 (d, J=2.4 Hz, 1H), 6.91 (dd, J=8.8, 2.4 Hz, 1H), 6.49 (d, J=2.4 Hz, 1H), 6.45 (dd, J=8.4, 2.4 Hz, 1H), 5.98 (s, 1H), 4.87 (d, J=4.4 Hz, 2H), 4.33 (t, J=7.6 Hz, 2H), 3.86 (s, 3H), 3.80 (s, 3H), 3.64-3.61 (m, 4H), 3.26-3.23 (m, 4H), 1.99-1.92 (m, 2H), 1.51 (s, 9H), 1.40-1.34 (m, 2H), 0.96 (t, J=7.2 Hz, 3H). 
     Example 49: Synthesis of Compound 51 
     
       
         
         
             
             
         
       
     
     To a mixture of tert-butyl 4-[2-butyl-4-[(2,4-dimethoxyphenyl)methylamino]pyrazolo[3,4-c]quinolin-7-yl]piperazine-1-carboxylate (0.2 g, 348.00 μmol, 1 eq) in DCM (20 mL) was added TFA (1.98 g, 17.40 mmol, 1.29 mL, 50 eq) in one portion at 25° C. The mixture was stirred at 50° C. for 36 hours. LCMS and HPLC showed the reaction was completed. The mixture was concentrated and purified by prep-HPLC (column: Nano-micro KROMASIL™ (Sigma-Aldrich) C18 100*30 mm Sum; mobile phase: [water (0.1% TFA)-ACN]; B %: 20%-55%, 10 min) to provide 2-butyl-7-piperazin-1-yl-pyrazolo[3,4-c]quinolin-4-amine (0.088 g, 200.71 μmol, 57.67% yield, TFA) as an off-white solid.  1 H NMR (DMSO-d 6 , 400 MHz) δ 9.01 (s, 2H), 8.88 (s, 1H), 7.96 (d, J=8.8 Hz, 1H), 7.23 (dd, J=8.8, 2.4 Hz, 1H), 7.11 (d, J=2.4 Hz, 1H), 4.49 (t, J=7.2 Hz, 2H), 3.45-3.44 (m, 4H), 3.35-3.29 (m, 4H), 1.98-1.90 (m, 2H), 1.36-1.27 (m, 2H), 0.93 (t, J=7.2 Hz, 3H). LCMS (ESI): mass calcd. for C 18 H 24 N 6  324.21, m/z found 325.3 [M+H] + . 
     Example 50: Synthesis of Compound 52 
     
       
         
         
             
             
         
       
     
     2-butyl-7-(piperazin-1-yl)-2H-pyrazolo[3,4-c]quinolin-4-amine was converted into tert-butyl 1-(4-(4-amino-2-butyl-2H-pyrazolo[3,4-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate in a 65% yield using the procedure described in Example 12. LC/MS [M+H] 893.56 (calculated); LC/MS [M+H] 893.82 (observed). 
     Example 51: Synthesis of Compound 53 
     
       
         
         
             
             
         
       
     
     tert-butyl 1-(4-(4-amino-2-butyl-2H-pyrazolo[3,4-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate was converted into 1-(4-(4-amino-2-butyl-2H-pyrazolo[3,4-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oic acid in a 92% yield using the procedure described in Example 16. The compound was used without further purification. 
     Example 52: Synthesis of Compound 54 
     
       
         
         
             
             
         
       
     
     1-(4-(4-amino-2-butyl-2H-pyrazolo[3,4-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oic acid was converted into 2,3,5,6-tetrafluorophenyl 1-(4-(4-amino-2-butyl-2H-pyrazolo[3,4-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate in a 46% yield using the procedure described in Example 17. LC/MS [M+H] 985.49 (calculated); LC/MS [M+H] 985.73 (observed). 
     Example 53: Synthesis of Compound 56 
     
       
         
         
             
             
         
       
     
     5-bromo-1H-indole was converted into 2,3,5,6-tetrafluorophenyl 1-(4-(4-amino-2-butyl-2H-pyrazolo[3,4-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate using the route described in Examples 45-53. LC/MS [M+H] 985.49 (calculated); LC/MS [M+H] 985.73 (observed). 
     Example 54: Synthesis of Compound 59 
     
       
         
         
             
             
         
       
     
     A 4 mL vial was charged with tert-butyl 1-azido-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate (0.077 mmol, 47 mg), triphenylphosphine (0.088 mmol, 23 mg) and 500 μL of anhydrous dichloromethane. The reaction was maintained at 30° C. for 90 min, at which point 3-cyanophenyl isocyanate (0.076 mmol, 11 mg) was added. After 45 min a solution containing 58.HCl* (0.077 mmol) and DIEA (0.345 mmol) in 750 mL of 2:1 DCM:DMF was added. This reaction was maintained for 2 h then concentrated under reduced pressure. The crude reaction was purified using reverse phase preparative HPLC utilizing a 25-75% gradient of acetonitrile:water containing 0.1% trifluoroacetic acid. The purified fractions were combined and lyophilized to provide 49.7 mg of the desired product tert-butyl (Z)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-cyanophenyl)amino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracont-34-enoate in 63% yield. LC/MS [M+H] 1023.61 (calculated); LC/MS [M+H] 1024.01 (observed). 
     Example 55: Synthesis of Compound 60 
     
       
         
         
             
             
         
       
     
     A 4 ml vial was charged with tert-butyl (Z)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-cyanophenyl)amino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracont-34-enoate (0.031 mmol, 32 mg), 1 mL 1,4-dioxane, and 1 mL 4M HCl in 1,4-dioxane. The reaction was stirred for 6 h, diluted with water (8 mL) and purified by reverse phase preparative HPLC utilizing a 25-75% gradient of acetonitrile:water containing 0.1% trifluoroacetic acid. The purified fractions were combined and lyophilized to afford 15.8 mg of the desired product (Z)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-cyanophenyl)amino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracont-34-enoic acid in 53% yield. LC/MS [M+H] 967.55 (calculated); LC/MS [M+H] 967.96 (observed). 
     Example 56: Synthesis of Compound 61 
     
       
         
         
             
             
         
       
     
     A 4 mL vial was charged with (Z)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-cyanophenyl)amino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracont-34-enoic acid (9.7 μmol, 9.4 mg) and 300 μL of DMF. To this vial was added 2,3,5,6-tetrafluorophenol (29 μmol, 4.1 mg) in 50 μL of DMF followed by 1-hydroxy-7-azabenzotriazole (9.4 μmol, 1.3 mg) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (29 μmol, 6.5 mg). The reaction was heat to 70° C. and stirred for 5 h, after which the reaction was cooled to 20° C., diluted with water (8 mL) and purified by reverse phase preparative HPLC utilizing a 25-75% gradient of acetonitrile:water containing 0.1% trifluoroacetic acid. The purified fractions were combined and lyophilized to afford 5.0 mg of the desired product 2,3,5,6-tetrafluorophenyl (Z)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-cyanophenyl)amino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracont-34-enoate in 46% yield. LC/MS [M+H] 1115.54 (calculated); LC/MS [M+H] 1115.98 (observed). 
     Example 57: Synthesis of Compound 63 
     
       
         
         
             
             
         
       
     
     A 20 mL vial was charged with oxalyl chloride (3.02 mmol, 260 μL) and 3 mL anhydrous DCM was cooled to −78° C. in a dry ice-acetone bath. A solution of DMSO (6.05 mmol, 429 μL) in 2 mL anhydrous DCM was added dropwise and the reaction was stirred for 1 h. A solution of tert-butyl 1-hydroxy-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-oate (1.01 mmol, 502 mg) in 2 mL anhydrous DCM was added dropwise and stirred for 20 min. Triethylamine (9.07 mmol, 1.23 mL) was added dropwise and the mixture was stirred for 40 min at −78° C. then warmed to room temperature over 30 min. The reaction was concentrated under reduced pressure and used immediately in the subsequent step. 
     Example 58: Synthesis of Compound 64 
     
       
         
         
             
             
         
       
     
     To a 20 ml vial containing crude tert-butyl 1-oxo-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-oate (1.01 mmol) and 2-(2-aminoethoxy)ethan-1-ol (2.37 mmol, 249 mg) in 10 mL anhydrous DCM was added triacetoxyborohydride (5.04 mmol, 1.07 g). The reaction was stirred for 5 h, then quenched with 1 mL of 10% K 2 CO 3 . The reaction was concentrated under reduced pressure. Purification by reverse phase flash chromatography using a 0-100% MeCN:water+0.1% TFA gradient afforded 276 mg of the desired product tert-butyl 1-hydroxy-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oate in 47% yield. LC/MS [M+H] 586.38 (calculated); LC/MS [M+H] 586.92 (observed). 
     Example 59: Synthesis of Compound 65 
     
       
         
         
             
             
         
       
     
     A 20 mL vial was charged with di-tert-butyl dicarbonate (1.39 mmol, 320 μL), tert-butyl 1-hydroxy-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oate (0.47 mmol, 276 mg), sodium bicarbonate (2.36 mmol, 200 mg), 4 mL THF, and 1 mL water. The reaction was stirred for 5 h, then concentrated under reduced pressure. Purification by reverse phase flash chromatography using a 0-100% MeCN:water+0.1% TFA gradient afforded 109 mg of the desired product tert-butyl 6-(tert-butoxycarbonyl)-1-hydroxy-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oate in 34% yield. LC/MS [M+H] 686.42 (calculated); LC/MS [M+H] 686.74 (observed). 
     Example 60: Synthesis of Compound 66 
     
       
         
         
             
             
         
       
     
     tert-butyl 6-(tert-butoxycarbonyl)-1-oxo-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oate was formed from tert-butyl 6-(tert-butoxycarbonyl)-1-hydroxy-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oate using Swern oxidation conditions according to Example 57. Crude material was carried forward to next step without purification. 
     Example 61: Synthesis of Compound 67 
     
       
         
         
             
             
         
       
     
     tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-6-(tert-butoxycarbonyl)-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oate was formed from tert-Butyl 6-(tert-butoxycarbonyl)-1-oxo-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oate using reductive amination conditions according to Example 58. Crude material was carried forward to next step without purification. 
     Example 62: Synthesis of Compound 68 
     
       
         
         
             
             
         
       
     
     1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oic acid was formed from tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-6-(tert-butoxycarbonyl)-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oate using Boc deprotection conditions according to Example 12. Crude material was carried forward to next step without purification. Purification by reverse phase flash chromatography using a 0-100% MeCN:water+0.1% TFA gradient afforded 109 mg of the desired product 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oic acid in 43% yield over three steps. LC/MS [M+H] 836.51 (calculated); LC/MS [M+H] 836.71 (observed). 
     Example 63: Synthesis of Compound 69 
     
       
         
         
             
             
         
       
     
     A 4 mL vial was charged with 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oic acid (0.025 mmol, 22 mg), 2,5-dioxopyrrolidin-1-yl 3-(2-(2-methoxyethoxy)ethoxy)propanoate (0.025 mmol, 6.4 mg), Hunigs base (0.067 mmol, 11.5 μL), 1-hydroxy-7-azabenzotriazole (0.014 mmol, 2 mg), 4-dimethylaminopyridine (0.016 mmol, 2 mg), and 500 μL DMF. The reaction was stirred at 45° C. for 4 h. Purification by reverse phase preparative HPLC utilizing a 25-75% gradient of acetonitrile:water+0.1% trifluoroacetic acid afforded 8.5 mg of the desired 12-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-11-oxo-2,5,8,15,18,21,24,27,30,33,36-undecaoxa-12-azanonatriacontan-39-oic acid in 34% yield. LC/MS [M+H] 1010.60 (calculated); LC/MS [M+H] 1010.76 (observed). 
     Example 64: Synthesis of Compound 70 
     
       
         
         
             
             
         
       
     
     2,3,5,6-tetrafluorophenyl 12-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-11-oxo-2,5,8,15,18,21,24,27,30,33,36-undecaoxa-12-azanonatriacontan-39-oate was formed from 12-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-11-oxo-2,5,8,15,18,21,24,27,30,33,36-undecaoxa-12-azanonatriacontan-39-oic acid following the procedure of Example 17. The procedure provided 7.7 mg of the desired 2,3,5,6-tetrafluorophenyl 12-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-11-oxo-2,5,8,15,18,21,24,27,30,33,36-undecaoxa-12-azanonatriacontan-39-oate in 79% yield. LC/MS [M+H] 1158.60 (calculated); LC/MS [M+H] 1158.88 (observed). 
     Example 65: Synthesis of Compound 71 
     
       
         
         
             
             
         
       
     
     33-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-32-oxo-2,5,8,11,14,17,20,23,26,29,36,39,42,45,48,51,54,57-octadecaoxa-33-azahexacontan-60-oic acid was formed from 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oic acid according to the procedure provided in Example 63. This procedure afforded 14.1 mg of the desired 33-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-32-oxo-2,5,8,11,14,17,20,23,26,29,36,39,42,45,48,51,54,57-octadecaoxa-33-azahexacontan-60-oic acid in 49% yield. LC/MS [M+H] 1318.79 (calculated); LC/MS [M+H] 1318.99 (observed). 
     Example 66: Synthesis of Compound 72 
     
       
         
         
             
             
         
       
     
     2,3,5,6-tetrafluorophenyl 33-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-32-oxo-2,5,8,11,14,17,20,23,26,29,36,39,42,45,48,51,54,57-octadecaoxa-33-azahexacontan-60-oate was formed from 33-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-32-oxo-2,5,8,11,14,17,20,23,26,29,36,39,42,45,48,51,54,57-octadecaoxa-33-azahexacontan-60-oic acid according to the procedure provided in Example 64. This procedure provided 11.9 mg of the desired 2,3,5,6-tetrafluorophenyl 33-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-32-oxo-2,5,8,11,14,17,20,23,26,29,36,39,42,45,48,51,54,57-octadecaoxa-33-azahexacontan-60-oate in 76% yield. LC/MS [M+H] 1466.78 (calculated); LC/MS [M+H] 1467.12 (observed). 
     Example 67: Synthesis of Compound 73 
     
       
         
         
             
             
         
       
     
     75-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-74-oxo-2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,78,81,84,87,90,93,96, 99-dotriacontaoxa-75-azadohectan-102-oic acid was formed from 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritriacontan-33-oic acid according to the procedure provided in Example 63. This procedure afforded 23.1 mg of the desired 75-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-74-oxo-2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,78,81,84,87,90,93,96, 99-dotriacontaoxa-75-azadohectan-102-oic acid in 54% yield. LC/MS [M+H] 1935.15 (calculated); LC/MS [M+H] 1935.30 (observed). 
     Example 68: Synthesis of Compound 74 
     
       
         
         
             
             
         
       
     
     2,3,5,6-tetrafluorophenyl 75-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-74-oxo-2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,78,81,84,87,90,93,96, 99-dotriacontaoxa-75-azadohectan-102-oate was formed from 75-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-74-oxo-2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,78,81,84,87,90,93,96, 99-dotriacontaoxa-75-azadohectan-102-oic acid according to the procedure provided in Example 64. This procedure provided 13.4 mg of the desired 2,3,5,6-tetrafluorophenyl 75-(2-(2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)ethoxy)ethyl)-74-oxo-2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,78,81,84,87,90, 93,96,99-dotriacontaoxa-75-azadohectan-102-oate in 86% yield. LC/MS [M+H] 2083.15 (calculated); LC/MS [M+H] 2083.31 (observed). 
     Example 69: Synthesis of Compound 76 
     
       
         
         
             
             
         
       
     
     7-bromoquinolin-4-ol (9.66 g, 43.11 mmol, 1 equiv.) was converted into 7-bromo-3-nitroquinolin-4-ol (7.46 g, 27.7 mmol, 64%) according to the procedure described in Example 2. LC/MS [M+H] 268.96/270.95 (calculated); LC/MS [M+H] 268.99/271.02 (observed). 
     Example 70: Synthesis of Compound 77 
     
       
         
         
             
             
         
       
     
     7-bromo-3-nitroquinolin-4-ol (7.46 g, 27.7 mmol, 1 equiv.) was converted into 7-bromo-4-chloro-3-nitroquinoline (6.88 g, 23.9 mmol, 86%) according to the procedure described in Example 3. LC/MS [M+H] 286.92/288.92 (calculated); LC/MS [M+H] 286.98/288.97 (observed). 
     Example 71: Synthesis of Compound 78 
     
       
         
         
             
             
         
       
     
     7-bromo-4-chloro-3-nitroquinoline (2.86 g, 9.95 mmol, 1 equiv.) was converted into 7-bromo-N-(2,4-dimethoxybenzyl)-3-nitroquinolin-4-amine (4.2 g, 10.0 mmol, 100%) according to the procedure described in Example 9. LC/MS [M+H] 418.04/420.04 (calculated); LC/MS [M+H] 418.19/420.16 (observed). 
     Example 72: Synthesis of Compound 79 
     
       
         
         
             
             
         
       
     
     7-bromo-N-(2,4-dimethoxybenzyl)-3-nitroquinolin-4-amine (4.2 g, 10.0 mmol, 1 equiv.) was suspended in acetonitrile (24 ml). Water (4 ml) was added, followed by nickel(II) chloride hexahydrate (0.48 g, 2 mmol, 0.2 equiv.). Sodium borohydride (1.52 g, 40.2 mmol, 4 equiv.) was added to the green suspension and the exothermic reaction was stirred for 30 minutes. The reaction mixture was filtered, concentrated, and purified by flash chromatography to give 7-bromo-N 4 -(2,4-dimethoxybenzyl)quinoline-3,4-diamine (2.15 g, 5.5 mmol, 55%). LC/MS [M+H] 388.07/390.06 (calculated); LC/MS [M+H] 388.22/390.21 (observed). 
     Example 73: Synthesis of Compound 80 
     
       
         
         
             
             
         
       
     
     7-bromo-N 4 -(2,4-dimethoxybenzyl)quinoline-3,4-diamine (2.15 g, 5.53 mmol, 1 equiv.) was dissolved in acetonitrile (25 ml). To the stirring solution were added triethyl orthovalerate (2.57 ml, 11.1 mmol, 2 equiv.) followed by iodine (0.140 g, 0.55 mmol, 0.1 equiv.). The reaction was stirred at room temperature until no starting material was observed by LCMS. The reaction mixture was concentrated, diluted in dichloromethane, and purified by flash chromatography to give 7-bromo-2-butyl-1-(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinoline (2.43 g, 5.3 mmol, 97%). LC/MS [M+H] 454.11/456.11 (calculated); LC/MS [M+H] 454.28/456.23 (observed). 
     Example 74: Synthesis of Compound 81 
     
       
         
         
             
             
         
       
     
     7-bromo-2-butyl-1-(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinoline (2.7 g, 5.94 mmol, 1 equiv.) was dissolved in 15 ml DCM. To the stirring reaction was added 4-chloroperoxybenzoic acid (4.39 g, 17.83 mmol, 3 equiv.). The reaction was stirred at room temperature and monitored by LCMS. Upon consumption of starting material, the reaction was quenched with 10% aqueous sodium carbonate, extracted with ethyl acetate, concentrated, and purified by flash chromatography to give 7-bromo-2-butyl-1-(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinoline 5-oxide (0.88 g, 1.87 mmol, 31%). LC/MS [M+H] 470.11/472.11 (calculated); LC/MS [M+H] 470.27/472.25 (observed). 
     Example 75: Synthesis of Compound 82 
     
       
         
         
             
             
         
       
     
     7-bromo-2-butyl-1-(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinoline 5-oxide (0.88 g, 1.87 mmol, 1 equiv.) was dissolved in dichloromethane (20 ml) and cooled on ice. Phosphoryl chloride (0.21 ml, 2.2 mmol, 1.2 equiv.) was added dropwise to the rapidly stirring solution, followed by N,N-dimethylformamide (0.072 ml, 0.94 mmol, 0.5 equiv.). After five minutes, the reaction was warmed to ambient temperature and monitored by LCMS. Upon consumption of starting material, the solution was washed with a mixture of ice and 10% aqueous sodium carbonate. The organic and aqueous layers were separated, and the aqueous layer extracted with dichloromethane (15 ml). The combined organic fractions were dried over sodium sulfate, filtered, and concentrated to provide 7-bromo-2-butyl-4-chloro-1-(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinoline as a brown foam (1.02 g, 2.09 mmol, 100%). LC/MS [M+H] 488.07/490.07 (calculated); LC/MS [M+H] 488.22/490.21 (observed). 
     Example 76: Synthesis of Compound 83 
     
       
         
         
             
             
         
       
     
     7-bromo-2-butyl-4-chloro-1-(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinoline (1 g, 2 mmol, 1 equiv.) was converted into 7-bromo-2-butyl-1-(2,4-dimethoxybenzyl)-N-(2,4-dimethoxyphenyl)-1H-imidazo[4,5-c]quinolin-4-amine using the procedure described in Example 21 (0.694 g, 1.12 mmol, 57%). LC/MS [M+H] 619.19/621.32 (calculated); LC/MS [M+H] 619.37/621.32 (observed). 
     Example 77: Synthesis of Compound 84 
     
       
         
         
             
             
         
       
     
     7-bromo-2-butyl-1-(2,4-dimethoxybenzyl)-N-(2,4-dimethoxyphenyl)-1H-imidazo[4,5-c]quinolin-4-amine (0.154 g, 0.25 mmol, 1 equiv.) and Pd(PPH 3 ) 4  (28.7 mg, 0.0025 mmol, 0.1 equiv.) were combined under dry dinitrogen. Cyanobutylzinc bromide (2.5 ml, 0.5 M in THF, 1.24 mmol, 5 equiv.) was added under dry dinitrogen and the reaction was heated to 75° C. After 30 minutes, another portion of cyanobutylzinc bromide was added (2.5 ml, 0.5 M in THF, 1.24 mmol, 5 equiv.) and the reaction allowed to stir for an additional 90 minutes. The solution was concentrated to a syrup and the crude material purified by flash chromatography to provide a mixture of the desired 5-(2-butyl-1-(2,4-dimethoxybenzyl)-4-((2,4-dimethoxyphenyl)amino)-1H-imidazo[4,5-c]quinolin-7-yl)pentanenitrile along with 2-butyl-1-(2,4-dimethoxybenzyl)-N-(2,4-dimethoxyphenyl)-1H-imidazo[4,5-c]quinolin-4-amine and residual solvent that was carried on as a crude mixture (0.288 g). LC/MS [M+H] 622.34 (calculated); LC/MS [M+H] 622.96 (observed). 
     Example 78: Synthesis of Compound 85 
     
       
         
         
             
             
         
       
     
     5-(2-butyl-1-(2,4-dimethoxybenzyl)-4-((2,4-dimethoxyphenyl)amino)-1H-imidazo[4,5-c]quinolin-7-yl)pentanenitrile (0.69 g, 1.1 mmol, 1 equiv.) was dissolved in methanol (20 ml) and cooled on ice. Nickel(II) chloride hexahydrate (0.053 g, 0.22 mmol, 0.2 equiv.) and Boc anhydride (0.51 ml, 2.22 mmol, 2 equiv.) were added to the stirring mixture. Sodium borohydride (1 g, 26.4 mmol, 23.8 equiv.) was added slowly in portions over 1 hour. The reaction was warmed and allowed to stand at ambient temperature for 1 hour, then concentrated. The crude material was taken up in ethyl acetate and washed with saturated sodium bicarbonate. The organic fraction was dried over sodium sulfate, filtered, concentrated, and then purified by flash chromatography to provide tert-butyl (5-(2-butyl-1-(2,4-dimethoxybenzyl)-4-((2,4-dimethoxyphenyl)amino)-1H-imidazo[4,5-c]quinolin-7-yl)pentyl)carbamate (0.265 g, 0.37 mmol, 33%). LC/MS [M+H] 726.42 (calculated); LC/MS [M+H] 726.64 (observed). 
     Example 79: Synthesis of Compound 86 
     
       
         
         
             
             
         
       
     
     2-butyl-N,1-bis(3,4-dimethylbenzyl)-7-(5-(methylamino)pentyl)-1H-imidazo[4,5-c]quinolin-4-amine was prepared from tert-Butyl (5-(2-butyl-1-(2,4-dimethoxybenzyl)-4-((2,4-dimethoxyphenyl)amino)-1H-imidazo[4,5-c]quinolin-7-yl)pentyl)carbamate (94.3 mg, 0.13 mmol, 1 equiv.) according to the procedure set forth in Example 31. LC/MS [M+H] 640.39 (calculated); LC/MS [M+H] 640.55 (observed). 
     Example 80: Synthesis of Compound 87 
     
       
         
         
             
             
         
       
     
     2-butyl-7-(5-(methylamino)pentyl)-1H-imidazo[4,5-c]quinolin-4-amine was prepared from 2-butyl-N,1-bis(3,4-dimethylbenzyl)-7-(5-(methylamino)pentyl)-1H-imidazo[4,5-c]quinolin-4-amine according to the procedure set forth in Example 32. LC/MS [M+H] 340.25 (calculated); LC/MS [M+H] 340.36 (observed). 
     Example 81: Synthesis of Compound 88 
     
       
         
         
             
             
         
       
     
     2-butyl-7-(5-(methylamino)pentyl)-1H-imidazo[4,5-c]quinolin-4-amine (50 mg, 0.15 mmol, 1 equiv.) was converted into tert-butyl 39-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)-34-methyl-4,7,10,13,16,19,22,25,28,31-decaoxa-34-azanonatriacontanoate using the procedure described in Example 12. LC/MS [M+H] 908.60 (calculated); LC/MS [M+H] 908.75 (observed). 
     Example 82: Synthesis of Compound 89 
     
       
         
         
             
             
         
       
     
     Tert-butyl 39-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)-34-methyl-4,7,10,13,16,19,22,25,28,31-decaoxa-34-azanonatriacontanoate was converted into 39-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)-34-methyl-4,7,10,13,16,19,22,25,28,31-decaoxa-34-azanonatriacontanoic acid (45 mg, 0.15 mmol, 33% from compound 87) using the procedure described in Example 16. LC/MS [M+H] 852.53 (calculated); LC/MS [M+H]852.75 (observed). 
     Example 83: Synthesis of Compound 90 
     
       
         
         
             
             
         
       
     
     39-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)-34-methyl-4,7,10,13,16,19,22,25,28,31-decaoxa-34-azanonatriacontanoic acid (45 mg, 0.053 mmol, 1 equiv.) was converted into 2,3,5,6-tetrafluorophenyl 39-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)-34-methyl-4,7,10,13,16,19,22,25,28,31-decaoxa-34-azanonatriacontanoate (28.5 mg, 0.053 mmol, 54%) according to the procedure described in Example 17. LC/MS [M+H] 1000.53 (calculated); LC/MS [M+H] 1000.72 (observed). 
     Example 84: Synthesis of Compound 92 
     
       
         
         
             
             
         
       
     
     5-bromoquinolin-4-ol was converted into 2,3,5,6-tetrafluorophenyl 39-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-9-yl)-34-methyl-4,7,10,13,16,19,22,25,28,31-decaoxa-34-azanonatriacontanoate using the route described in Examples 69-83. LC/MS [M+H] 1000.53 (calculated); LC/MS [M+H] 1000.94 (observed). 
     Example 85: Synthesis of Compound 93 
     
       
         
         
             
             
         
       
     
     Compound 39 was converted into 2,3,5,6-tetrafluorophenyl 39-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)-34-methyl-4,7,10,13,16,19,22,25,28,31-decaoxa-34-azanonatriacontanoate using the route described in Examples 77-83. LC/MS [M+H] 1000.53 (calculated); LC/MS [M+H] 1000.92 (observed). 
     Example 86: Synthesis of Compound 94 
     
       
         
         
             
             
         
       
     
     5-(2-butyl-1-(2,4-dimethoxybenzyl)-4-((2,4-dimethoxybenzyl)amino)-1H-imidazo[4,5-c]quinolin-7-yl)pentanenitrile (0.288 g, 0.46 mmol, 1 equiv.) was dissolved in 4 ml dry THF. Lithium aluminum hydride (0.088 g, 2.3 mmol, 5 equiv.) was added and the exothermic reaction stirred at ambient temperature for 1 hour. The reaction was quenched with 0.5 ml saturated NaHCO 3 , diluted with 10 ml THF, and precipitate removed by centrifugation. The resulting solution was concentrated and purified by HPLC to provide 7-(5-aminopentyl)-2-butyl-N,1-bis(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinolin-4-amine (0.101 g, 0.16 mmol, 35%). LC/MS [M+H] 626.37 (calculated); LC/MS [M+H] 626.56 (observed). 
     Example 87: Synthesis of Compound 95 
     
       
         
         
             
             
         
       
     
     7-(5-aminopentyl)-2-butyl-N,1-bis(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinolin-4-amine (23.2 mg, 0.037 mmol, 1 equiv.) was suspended in dry N,N-dimethylformamide (2 ml). Diisopropylamine (0.065 ml, 0.37 mmol, 10 equiv.) was added, followed by (tert-butoxycarbonyl)(sulfo)-D-alanine (0.020 g, 0.074 mmol, 2 equiv.) and HATU (0.049 g, 0.13 mmol, 3.5 equiv.). The reaction was stirred at 50° C. for 30 minutes, then diluted with water and purified by HPLC to provide (R)-2-((tert-butoxycarbonyl)amino)-3-((5-(2-butyl-1-(2,4-dimethoxybenzyl)-4-((2,4-dimethoxybenzyl)amino)-1H-imidazo[4,5-c]quinolin-7-yl)pentyl)amino)-3-oxopropane-1-sulfonic acid (28.3 mg, 0.032 mmol, 87%). LC/MS [M+H] 877.42 (calculated); LC/MS [M+H] 877.59 (observed). 
     Example 88: Synthesis of Compound 96 
     
       
         
         
             
             
         
       
     
     (R)-2-((tert-butoxycarbonyl)amino)-3-((5-(2-butyl-1-(2,4-dimethoxybenzyl)-4-((2,4-dimethoxybenzyl)amino)-1H-imidazo[4,5-c]quinolin-7-yl)pentyl)amino)-3-oxopropane-1-sulfonic acid (28.3 mg, 0.032 mmol) was converted into (R)-2-amino-3-((5-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)pentyl)amino)-3-oxopropane-1-sulfonic acid (15.8 mg, 0.033 mmol, 100%) according to the procedure described in Example 32. LC/MS [M+H] 877.42 (calculated); LC/MS [M+H] 877.59 (observed). 
     Example 89: Synthesis of Compound 97 
     
       
         
         
             
             
         
       
     
     (R)-2-amino-3-((5-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)pentyl)amino)-3-oxopropane-1-sulfonic acid (15.8 mg, 0.033 mmol) was dissolved in dry N,N-dimethylformamide (1 ml). A solution of 31-((2,5-dioxocyclopentyl)oxy)-31-oxo-4,7,10,13,16,19,22,25,28-nonaoxahentriacontanoic acid (22.3 mg, 0.036 mmol, 1.1 equiv.) in N,N-dimethylformamide (1 ml) was added, followed by diisopropylethylamine (0.01 ml, 0.057 mmol, 1.7 equiv.). The reaction was stirred at 60° C. until complete by LCMS, then purified directly by HPLC to provide (R)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)-33-((hydroxy(l1-oxidaneyl)(oxo)-15-sulfaneyl)methyl)-31,34-dioxo-4,7,10,13,16,19,22,25,28-nonaoxa-32,35-diazatetracontanoic acid (22 mg, 0.023 mmol, 68% yield.). LC/MS [M+H] 973.48 (calculated); LC/MS [M+H] 973.76 (observed). 
     Example 90: Synthesis of Compound 98 
     
       
         
         
             
             
         
       
     
     (R)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)-33-((hydroxy(l1-oxidaneyl)(oxo)-15-sulfaneyl)methyl)-31,34-dioxo-4,7,10,13,16,19,22,25,28-nonaoxa-32,35-diazatetracontanoic acid (22 mg, 0.023 mmol) was converted into 2,3,5,6-tetrafluorophenyl (R)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)-33-((hydroxy(l1-oxidaneyl)(oxo)-15-sulfaneyl)methyl)-31,34-dioxo-4,7,10,13,16,19,22,25,28-nonaoxa-32,35-diazatetracontanoate (14.2 mg, 0.013 mmol, 56%) according to the procedure described in Example 17. LC/MS [M+H] 1121.47 (calculated); LC/MS [M+H] 1121.68 (observed). 
     Example 91: Synthesis of Compound 99 
     
       
         
         
             
             
         
       
     
     2-butyl-7-(5-(methylamino)pentyl)-1H-imidazo[4,5-c]quinolin-4-amine (0.1 g, 0.29 mmol, 1 equiv.) was dissolved in dry N,N-dimethylformamide (2 ml). Sodium triacetoxyborohydride (0.250 g, 1.17 mmol, 4 equiv.) was added, followed by formaldehyde (0.048 ml, 37% by mass, 0.597 mmol, 2 equiv.). After 15 minutes, the reaction was diluted with water and purified by HPLC to provide 2-butyl-7-(5-(dimethylamino)pentyl)-1H-imidazo[4,5-c]quinolin-4-amine (0.044 g, 0.12 mmol, 42%). LC/MS [M+H] 354.27 (calculated); LC/MS [M+H] 354.40 (observed). 
     Example 92: Synthesis of Compound 100 
     
       
         
         
             
             
         
       
     
     A 20 mL vial was charged with tert-butyl 1-hydroxy-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate (0.4 mmol, 234 mg), triethylamine (0.8 mmol, 111 μL), methanesulfonyl chloride (0.44 mmol, 34 μL), and 7 mL Toluene. The reaction was heated to 60° C. for 90 min. The reaction was cooled to room temperature, filtered through diatomaceous earth, and concentrated under reduced pressure. To this concentrated crude mixture was added potassium iodide and 8 mL acetone. The reaction was heated to 50° C. for 12 h. The reaction was cooled to room temperature, filtered through diatomaceous earth, and concentrated under reduced pressure. The crude material was dissolved in dichloromethane, filtered through diatomaceous earth, concentrated under reduced pressure and used in the subsequent reaction without further purification. 2-butyl-7-(5-(dimethylamino)pentyl)-1H-imidazo[4,5-c]quinolin-4-amine (29.8 mg, 0.084 mmol, 1 equiv.) was dissolved in dry N,N-dimethylformamide (1 ml). Sodium bicarbonate (0.1 g) was added and the suspension stirred at ambient temperature for 5 minutes. tert-Butyl 1-iodo-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate was added as a solution in dry N,N-dimethylformamide (0.5 ml), and the reaction was stirred at 60° C. The reaction solution was filtered, diluted with water, and purified by HPLC. The resulting material was taken up in 0.05 ml trifluoroacetic acid and allowed to stand at ambient temperature for 30 minutes before concentration and purification by HPLC to provide N-(5-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)pentyl)-32-carboxy-N,N-dimethyl-3,6,9,12,15,18,21,24,27,30-decaoxadotriacontan-1-aminium (14.3 mg, 0.016 mmol, 20%). LC/MS [M+H] 866.55 (calculated); LC/MS [M+H] 866.72 (observed). 
     Example 93: Synthesis of Compound 101 
     
       
         
         
             
             
         
       
     
     N-(5-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)pentyl)-32-carboxy-N,N-dimethyl-3,6,9,12,15,18,21,24,27,30-decaoxadotriacontan-1-aminium (14.3 mg, 0.016 mmol) was converted to N-(5-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)pentyl)-N,N-dimethyl-33-oxo-33-(2,3,5,6-tetrafluorophenoxy)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-1-aminium (11.4 mg, 0.011 mmol, 68%) using the procedure described in Example 17. LC/MS [M+H] 1014.54 (calculated); LC/MS [M+H] 1014.76 (observed). 
     Example 94: Synthesis of Compound 102 
     
       
         
         
             
             
         
       
     
     7-bromo-2-butyl-N,1-bis(2,4-dimethoxybenzyl)-1H-imidazo[4,5-c]quinolin-4-amine was converted into tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate according to the procedures described in Examples 23-27. LC/MS [M+H] 893.56 (calculated); LC/MS [M+H] 893.79. 
     Example 95: Synthesis of Compound 103 
     
       
         
         
             
             
         
       
     
     tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate was converted to 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oic acid according to the procedure set forth in Example 16. LC/MS [M+H] 837.49 (calculated); LC/MS [M+H] 837.84 (observed). 
     Example 96: Synthesis of Compound 104 
     
       
         
         
             
             
         
       
     
     1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oic acid was converted to 2,3,5,6-tetrafluorophenyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-7-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-oate according to the procedure set forth in Example 17. LC/MS [M+H] 985.49 (calculated); [M+H] 985.71 (observed). 
     Example 97: Synthesis of Compound 105 
     
       
         
         
             
             
         
       
     
     1-(4-aminobutyl)-2-butyl-1H-imidazo[4,5-c]quinolin-4-amine was converted to (R)-3-((4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl)amino)-2-((tert-butoxycarbonyl)amino)-3-oxopropane-1-sulfonic acid according to the procedure described in Example 87. LC/MS [M+H] 563.27 (calculated); LC/MS [M+H] 563.43 (observed). 
     Example 98: Synthesis of Compound 106 
     
       
         
         
             
             
         
       
     
     (R)-3-((4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl)amino)-2-((tert-butoxycarbonyl)amino)-3-oxopropane-1-sulfonic acid was converted to (R)-2-amino-3-((4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl)amino)-3-oxopropane-1-sulfonic acid according to the procedure described in Example 88. LC/MS [M+H] 463.21 (calculated); LC/MS [M+H] 463.38 (observed). 
     Example 99: Synthesis of Compound 107 
     
       
         
         
             
             
         
       
     
     (R)-2-amino-3-((4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl)amino)-3-oxopropane-1-sulfonic acid was converted to (R)-45-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-37,40-dioxo-39-(sulfomethyl)-4,7,10,13,16,19,22,25,28,31,34-undecaoxa-38,41-diazapentatetracontanoic acid according to the procedure described in Example 89. LC/MS [M+H] 1047.52 (calculated); LC/MS [M+H] 1047.72 (observed). 
     Example 100: Synthesis of Compound 108 
     
       
         
         
             
             
         
       
     
     (R)-45-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-37,40-dioxo-39-(sulfomethyl)-4,7,10,13,16,19,22,25,28,31,34-undecaoxa-38,41-diazapentatetracontanoic acid was converted to (R)-2-((4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl)carbamoyl)-4,40-dioxo-40-(2,3,5,6-tetrafluorophenoxy)-7,10,13,16,19,22,25,28,31,34,37-undecaoxa-3-azatetracontane-1-sulfonic acid according to the procedure described in Example 17. LC/MS [M+H] 1195.51 (calculated); LC/MS [M+H] 1195.73 (observed). 
     Example 101: Synthesis of Compound 109 
     
       
         
         
             
             
         
       
     
     2-butyl-8-(piperazin-1-yl)-1H-imidazo[4,5-c]quinolin-4-amine was converted into tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18-hexaoxahenicosan-21-oate according to the procedure described in Example 12. LC/MS [M+H] 717.45 (calculated); LC/MS [M+H] 717.75 (observed). 
     Example 102: Synthesis of Compound 110 
     
       
         
         
             
             
         
       
     
     Tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18-hexaoxahenicosan-21-oate was converted into 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18-hexaoxahenicosan-21-oic acid according to the procedure described in Example 16. LC/MS [M+H] 661.39 (calculated); LC/MS [M+H] 661.60 (observed). 
     Example 103: Synthesis of Compound 111 
     
       
         
         
             
             
         
       
     
     1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18-hexaoxahenicosan-21-oic acid was converted into 2,3,5,6-tetrafluorophenyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18-hexaoxahenicosan-21-oate according to the procedure described in Example 17. LC/MS [M+H] 809.39 (calculated); LC/MS [M+H] 809.62 (observed). 
     Example 104: Synthesis of Compound 112 
     
       
         
         
             
             
         
       
     
     2-butyl-8-(piperazin-1-yl)-1H-imidazo[4,5-c]quinolin-4-amine was converted into tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36-dodecaoxanonatriacontan-39-oate according to the procedure described in Example 12. LC/MS [M+H] 981.61 (calculated); LC/MS [M+H] 981.86 (observed). 
     Example 105: Synthesis of Compound 113 
     
       
         
         
             
             
         
       
     
     Tert-butyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36-dodecaoxanonatriacontan-39-oate was converted into 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36-dodecaoxanonatriacontan-39-oic acid according to the procedure described in Example 16. The compound was used without further purification. 
     Example 106: Synthesis of Compound 114 
     
       
         
         
             
             
         
       
     
     1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36-dodecaoxanonatriacontan-39-oic acid was converted into 2,3,5,6-tetrafluorophenyl 1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36-dodecaoxanonatriacontan-39-oate according to the procedure described in Example 17. LC/MS [M+H] 1073.54 (calculated); LC/MS [M+H] 1073.81 (observed). 
     Example 107: Synthesis of Compound 115 
     
       
         
         
             
             
         
       
     
     A 4 mL vial was charged with 1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-1H-pyrrole-2,5-dione (0.089 mmol, 88 mg), 2,4,6-collidine (0.27 mmol, 36 μL) and 800 μL. To this was added a solution of compound 44 (0.098 mmol, 33 mg), HOAT (0.014 mmol, 2 mg), DIPEA (0.20 mmol, 36 μL) in 100 μL DMF. The reaction was stirred for 4 h and then purified by reverse phase preparative HPLC utilizing a 25-75% gradient of acetonitrile:water containing 0.1% trifluoroacetic acid. The purified fractions were combined and lyophilized to afford 46.7 mg of the desired product Compound 115 in 50% yield. LC/MS [M+H] 1047.60 (calculated); LC/MS [M+H] 1048.01 (observed). 
     Example 108: Synthesis of Compound 116 
     
       
         
         
             
             
         
       
     
     A 4 mL vial was charged with 1-(4-aminobutyl)-2-butyl-1H-imidazo[4,5-c]quinolin-4-amine* (0.13 mmol, 44 mg), formaldehyde (0.38 mmol, 31 μL of 37% solution), sodium triacetoxyborohydride (0.65 mmol, 135 mg), and 1 mL THF. Let stir for 1 h then quench with 100 μL of 10% K 2 CO 3 . The reaction was concentrated under reduced pressure and purified by reverse phase preparative HPLC utilizing a 25-75% gradient of acetonitrile:water containing 0.1% trifluoroacetic acid. The purified fractions were combined and lyophilized to afford 24.5 mg of the desired product 2-butyl-1-(4-(methylamino)butyl)-1H-imidazo[4,5-c]quinolin-4-amine in 57% yield. LC/MS [M+H] 340.25 (calculated); LC/MS [M+H] 340.40 (observed). 
     Example 109: Synthesis of Compound 117 
     
       
         
         
             
             
         
       
     
     A 20 mL vial was charged with tert-butyl 1-hydroxy-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-oate (0.4 mmol, 200 mg), triethylamine (0.8 mmol, 111 μL), methanesulfonyl chloride (0.44 mmol, 34 μL), and 7 mL Toluene. The reaction was heated to 60° C. for 90 min. The reaction was cooled to room temperature, filtered through diatomaceous earth, and concentrated under reduced pressure. To this concentrated crude mixture was added potassium iodide and 8 mL acetone. The reaction was heated to 50° C. for 12 h. The reaction was cooled to room temperature, filtered through diatomaceous earth, and concentrated under reduced pressure. The crude material was dissolved in dichloromethane, filtered through diatomaceous earth, concentrated under reduced pressure and used in the subsequent reaction without further purification. A 4 mL vial was charged with tert-butyl 1-iodo-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-oate (0.097 mmol, 59 mg), 2-butyl-1-(4-(methylamino)butyl)-1H-imidazo[4,5-c]quinolin-4-amine (0.081 mmol, 27.4 mg), K 2 CO 3  (0.16 mmol, 22.3 mg), acetonitrile (700 μL), and water (50 μL). The reaction was heated to 70° C. and stirred for 3 h. The reaction was purified by reverse phase preparative HPLC utilizing a 25-75% gradient of acetonitrile:water containing 0.1% trifluoroacetic acid. The purified fractions were combined and lyophilized to afford 22.2 mg of the desired product tert-butyl 32-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-28-methyl-4,7,10,13,16,19,22,25-octaoxa-28-azadotriacontanoate in 34% yield. LC/MS [M+H] 820.54 (calculated); LC/MS [M+H] 820.75 (observed). 
     Example 110: Synthesis of Compound 118 
     
       
         
         
             
             
         
       
     
     N-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl)-N,N,29,29-tetramethyl-27-oxo-3,6,9,12,15,18,21,24,28-nonaoxatriacontan-1-aminium was converted into N-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl)-26-carboxy-N,N-dimethyl-3,6,9,12,15,18,21,24-octaoxahexacosan-1-aminium according to the procedure described in Example 16. Compound was in subsequent step without further purification. 
     Example 111: Synthesis of Compound 119 
     
       
         
         
             
             
         
       
     
     N-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl)-26-carboxy-N,N-dimethyl-3,6,9,12,15,18,21,24-octaoxahexacosan-1-aminium was converted into N-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl)-N,N-dimethyl-27-oxo-27-(2,3,5,6-tetrafluorophenoxy)-3,6,9,12,15,18,21,24-octaoxaheptacosan-1-aminium according to the procedure described in Example 17. LC/MS 912.47 [M+H] (calculated); LC/MS [M+H]912.70 (observed). 
     Example 112: Synthesis of Compound 120 
     
       
         
         
             
             
         
       
     
     2-butyl-8-(piperazin-1-yl)-1H-imidazo[4,5-c]quinolin-4-amine was converted to (R)-3-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-2-((tert-butoxycarbonyl)amino)-3-oxopropane-1-sulfonic acid according to the procedure described in Example 87. LC/MS [M+H] 576.26 (calculated); LC/MS [M+H] 576.44 (observed). 
     Example 113: Synthesis of Compound 121 
     
       
         
         
             
             
         
       
     
     (R)-3-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-2-((tert-butoxycarbonyl)amino)-3-oxopropane-1-sulfonic acid was converted to (R)-2-amino-3-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3-oxopropane-1-sulfonic acid according to the procedure described in Example 88. The compound was used in the subsequent step without further purification. 
     Example 114: Synthesis of Compound 122 
     
       
         
         
             
             
         
       
     
     (R)-2-amino-3-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-3-oxopropane-1-sulfonic acid was converted to (R)-1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-1,4-dioxo-2-(sulfomethyl)-7,10,13,16,19,22,25,28,31,34,37-undecaoxa-3-azatetracontan-40-oic acid according to the procedure described in Example 89. LC/MS [M+H] 1060.51 (calculated); LC/MS [M+H]1060.73 (observed). 
     Example 115: Synthesis of Compound 123 
     
       
         
         
             
             
         
       
     
     (R)-1-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazin-1-yl)-1,4-dioxo-2-(sulfomethyl)-7,10,13,16,19,22,25,28,31,34,37-undecaoxa-3-azatetracontan-40-oic acid was converted to (R)-2-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-8-yl)piperazine-1-carbonyl)-4,40-dioxo-40-(2,3,5,6-tetrafluorophenoxy)-7,10,13,16,19,22,25,28,31,34,37-undecaoxa-3-azatetracontane-1-sulfonic acid according to the procedure described in Example 17. LC/MS [M+H] 1208.51 (calculated); LC/MS [M+H] 1208.74 (observed). 
     Example 116: Synthesis of Compound 124 
     
       
         
         
             
             
         
       
     
     A 4 mL vial was charged with tert-butyl (Z)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-cyanophenyl)amino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracont-34-enoate (0.017 mmol, 17 mg), 22 mg of 10% Pd/C, and 1.1 mL of ethanol. The vial was evacuated and backfilled thrice with hydrogen gas. The reaction was stirred for 4 h, filtered through diatomaceous earth, and concentrated under reduced pressure. The crude reaction was dissolved in dichloromethane and concentrated under reduced pressure. To the vial containing the crude reaction mixture was added 2,5-dioxopyrrolidin-1-yl 2,5,8,11,14,17,20,23,26,29-decaoxadotriacontan-32-oate (0.017 mmol, 11 mg), Hunigs base (0.069 mmol, 12 μL) and 300 μL DMF. The reaction was stirred for 4 h. The crude reaction was purified using reverse phase preparative HPLC utilizing a 25-75% gradient of acetonitrile:water containing 0.1% trifluoroacetic acid. The purified fractions were combined and lyophilized to afford 7.1 mg of the desired product tert-butyl (E)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-(32-oxo-2,5,8,11,14,17,20,23,26,29-decaoxa-33-azatetratriacontan-34-yl)phenyl)imino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracontanoate in 28% yield over two steps. LC/MS [M+H] 1509.92 (calculated); LC/MS [M+H] 1510.13 (observed). 
     Example 117: Synthesis of Compound 125 
     
       
         
         
             
             
         
       
     
     A 4 mL vial was charged with Tert-butyl (E)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-(32-oxo-2,5,8,11,14,17,20,23,26,29-decaoxa-33-azatetratriacontan-34-yl)phenyl)imino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracontanoate (4.7 μmol, 7.1 mg) and 800 μL of 4M HCl in 1,4-dioxane. The reaction was stirred for 4 h then concentrated under reduced pressure. The crude reaction was dissolved in 1 mL Toluene and concentrated under reduced pressure thrice. The crude material was taken on without further purification. 
     Example 118: Synthesis of Compound 126 
     
       
         
         
             
             
         
       
     
     A 4 mL vial was charged with (E)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-(32-oxo-2,5,8,11,14,17,20,23,26,29-decaoxa-33-azatetratriacontan-34-yl)phenyl)imino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracontanoic acid (4.7 μmol), 2,3,5,6-tetrafluorophenol (9.4 μmol, 1.6 mg), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (26 μmol, 5 mg), 2,4,6-collidine (60 μmol, 8 μL) and 100 μL of DMF. The reaction was stirred for 2 h then purified by reverse phase preparative HPLC utilizing a 25-75% gradient of acetonitrile:water containing 0.1% trifluoroacetic acid. The purified fractions were combined and lyophilized to afford 3.4 mg of the desired product 2,3,5,6-tetrafluorophenyl (E)-40-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-35-((3-(32-oxo-2,5,8,11,14,17,20,23,26,29-decaoxa-33-azatetratriacontan-34-yl)phenyl)imino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracontanoate 2,5,8,11,14,17,20,23,26,29-decaoxa-33-azatetratriacontan-34-yl)phenyl)imino)-4,7,10,13,16,19,22,25,28,31-decaoxa-34,36-diazatetracontanoic acid in 45% yield over two steps. LC/MS [M+H] 1601.85 (calculated); LC/MS [M+H] 1602.07 (observed). 
     Example 119: Synthesis of Immunoconjugate A 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate A with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich, St. Louis, Mo.). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 7 molar equivalents of Compound 44. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate A was separated from reactants by running over two successive G-25 desalting columns equilibrated in phosphate buffered saline (PBS) at pH 7.2. Adjuvant-antibody ratio (DAR) was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation, Milford, Mass.) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate A had a DAR of 2.5. 
     Example 120: Synthesis of Immunoconjugate B 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate B with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 7.4 molar equivalents of Compound 93. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate B was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DA-R was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G-2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate B had a DAR of 2.56. 
     Example 121: Synthesis of Immunoconjugate C 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate C with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 83, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 7.5 molar equivalents of Compound 56. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate C was separated from reactants by running over two successive (3-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G 2 -XS TOF mass spectrometer (Waters Corporation). Immunoconjugate C had a DAR of 2.65. 
     Example 122: Synthesis of Immunoconjugate D 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate D with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 83, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 8.5 molar equivalents of Compound 104. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate D was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate D had a DAR of 2.26. 
     Example 123: Synthesis of Immunoconjugate E 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate E with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 8 molar equivalents of Compound 18. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate E was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate E had a DAR of 17. 
     Example 124: Synthesis of Immunoconjugate F 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate F with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 8 molar equivalents of Compound 92. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate F was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate F had a DAR of 2.8. 
     Example 125: Synthesis of Immunoconjugate G 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate G with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 7.8 molar equivalents of Compound 54. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate G was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate G had a DAR of 2.39. 
     Example 126: Synthesis of Immunoconjugate H 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate H with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 13 molar equivalents of Compound 29. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate H was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G 2 -XS TOF mass spectrometer (Waters Corporation). Immunoconjugate H had a DAR of 1.76. 
     Example 127: Synthesis of Immunoconjugate I 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate I with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 8 molar equivalents of Compound 35. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate I was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate I had a DAR of 2.08. 
     Example 128: Synthesis of Immunoconjugate J 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate J with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was incubated in a solution of 20 mM TCEP in borate buffer at a pH of 8.3 at room temperature for one hour. The TCEP was removed using 10 kDa cut-off ZEBA™ column (Thermo Fisher Scientific, Waltham, Mass.) and the resulting trastuzumab was buffer-exchanged into PBS at a pH of 7.2. The thiols were subsequently quantified with a 4,4′-dithiodipyridine (4,4′-DTDP) assay. The reduced antibody (10 to 15 mg/ml) was mixed with 8, 15, 24, or 28 molar equivalents of Compound 115 for 1.5 hours at room temperature at 3.0× molar excess of the measured thiols from the assay. N-acetyl cysteine was added at 10× measured thiol concentration and the reaction was incubated at room temperature for one hour. The remnants of Compound 115 and N-acetyl cysteine were removed by buffer exchange (using 5 ml 40 kDa cut-off ZEBA™ column (Thermo Fisher Scientific)) with phosphate-buffered saline at a pH of 7.2. DAR was determined by an UV-Vis spectrophotometer. Immunoconjugate J had a DAR of 1.9, 5, 7, and 8 for 8, 15, 24, and 28 molar equivalents of Compound 115, respectively. 
     Example 129: Synthesis of Immunoconjugate L 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate L with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 6.5 molar equivalents of Compound 108. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate L was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2, DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate L had a DAR of 2.72. 
     Example 130: Synthesis of Immunoconjugate M 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate M with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 7.4 molar equivalents of Compound 123. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate M was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G 2 -XS TOF mass spectrometer (Waters Corporation). Immunoconjugate M had a DAR of 2.52. 
     Example 131: Synthesis of Immunoconjugate N 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate N with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 7.8 molar equivalents of Compound 70. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate N was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOY mass spectrometer (Waters Corporation). Immunoconjugate N had a DAR of 2.8. 
     Example 132: Synthesis of Immunoconjugate O 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate O with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPIADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 7.7 molar equivalents of Compound 72. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate O was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate O had a DAR of 3.21. 
     Example 133: Synthesis of Immunoconjugate P 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate P with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered, Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 8.5 molar equivalents of Compound 111. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate P was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate P had a DAR of 2.37. 
     Example 134: Synthesis of Immunoconjugate Q 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate Q with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 6 molar equivalents of Compound 74. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate Q was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate Q had a DAR of 2.77. 
     Example 135: Synthesis of Immunoconjugate R 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate R with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 83, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 6 molar equivalents of Compound 90. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate R was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate R had a DAR of 1.98. 
     Example 136: Synthesis of Immunoconjugate S 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate S with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with S molar equivalents of Compound 61. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate S was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate S had a DAR of 2.58. 
     Example 137: Synthesis of Immunoconjugate T 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate T with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 6.5 molar equivalents of Compound 126. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate T was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate T had a DAR of 2.73. 
     Example 138: Synthesis of Immunoconjugate U 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate U with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 7 molar equivalents of Compound 98. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate U was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate U had a DAR of 2.64. 
     Example 139: Synthesis of Immunoconjugate V 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate V with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 6 molar equivalents of Compound 114. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate V was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate V had a DAR of 2.15. 
     Example 140: Synthesis of Immunoconjugate K1 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate K 1  with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered. Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 30 molar equivalents of Compound 119. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate K1 was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate K1 had a DAR of 1.73. 
     Example 141: Synthesis of Immunoconjugate R1 
     
       
         
         
             
             
         
       
     
     This example demonstrates the synthesis of Immunoconjugate R1 with trastuzumab as the antibody construct (Tras). 
     Trastuzumab was buffer exchanged into the conjugation buffer containing 100 mM boric acid, 50 mM sodium chloride, 1 mM ethylenediaminetetraacetic acid at pH 8.3, using G-25 SEPHADEX™ desalting columns (Sigma-Aldrich). The eluates were then each adjusted to 6 mg/ml using the buffer and sterile filtered, Trastuzumab at 6 mg/ml was pre-warmed to 30° C. and rapidly mixed with 8 molar equivalents of Compound 101. The reaction was allowed to proceed for 16 hours at 30° C. and Immunoconjugate R1 was separated from reactants by running over two successive G-25 desalting columns equilibrated in PBS at pH 7.2. DAR was determined by liquid chromatography mass spectrometry analysis using a C4 reverse phase column on an ACQUITY™ UPLC H-class (Waters Corporation) connected to a XEVO™ G2-XS TOF mass spectrometer (Waters Corporation). Immunoconjugate R1 had a DAR of 2.2. 
     Example 142. Assessment of Immunoconjugate Activity In Vitro 
     This example shows that Immunoconjugates A-I, L-V, K1, and R1 are effective at eliciting myeloid activation, and therefore are useful for the treatment of cancer. 
     Isolation of Human Antigen Presenting Cells. Human myeloid antigen presenting cells (APCs) were negatively selected from human peripheral blood obtained from healthy blood donors (Stanford Blood Center, Palo Alto, Calif.) by density gradient centrifugation using a ROSETTESEP™ Human Monocyte Enrichment Cocktail (Stem Cell Technologies, Vancouver, Canada) containing monoclonal antibodies against CD14, CD16, CD40, CD86, CD123, and HLA-DR. Immature APCs were subsequently purified to &gt;97% purity via negative selection using an EASYSEP™ Human Monocyte Enrichment Kit (Stem Cell Technologies) without CD16 depletion containing monoclonal antibodies against CD14, CD16, CD40, CD86, CD123, and HLA-DR. 
     Preparation of Tumor Cells. Three tumor cell lines were used: HCC1954, JIMT-1, and COLO 205. HCC1954 (American Type Culture Collection (ATCC), Manassas, Va.) was derived from a primary stage IIA, grade 3 invasive ductal carcinoma with no lymph node metastases. HCC1954 is positive for the epithelial cell specific marker Epithelial Glycoprotein 2 and for cytokeratin 19, and is negative for expression of estrogen receptor (ER) and progesterone receptor (PR). HCC1954 overexpresses HER2 (as determined by enzyme-linked immunosorbent assay (ELISA)) with a relatively “high” level of overexpression. JIMT-1 (DSMZ, Braunschweig, Germany) was derived from the pleural effusion of a woman with ductal breast cancer (grade 3 invasive, stage IIB) following postoperative radiation. JIMT-1 overexpresses HER2 at what is considered to be a “medium” level of overexpression, but is insensitive to HER2-inhibiting drugs (e.g., trastuzumab). COLO 205 (ATCC) was derived from the ascites fluid of man with carcinoma of the colon. COLO 205 expresses carcinoembryonic antigen (CEA), keratin, interleukin 10 (IL-10), and is considered to overexpress HER2 at a relatively “low” level of overexpression. 
     Tumor cells from each cell line were separately re-suspended in PBS with 0.1% fetal bovine serum (FBS) at 1 to 10×10 6  cells/mL. Cells were subsequently incubated with 2 μM carboxyfluorescein succinimidyl ester (CFSE) to yield a final concentration of 1 μM. The reaction was quenched after 2 minutes via the addition of 10 mL complete medium with 10% FBS and washed twice with complete medium. Cells were either fixed in 2% paraformaldehyde and washed three times with PBS or left viable prior to use. 
     APC-Tumor Co-cultures. 2×10 5  APCs were incubated with (e.g.,  FIG. 1A-1I ) or without (e.g.,  FIG. 2A-22D ) CFSE-labeled tumor cells between a 5:1 and 10:1 effector to target (tumor) cell ratio in 96-well plates (Corning, Corning, N.Y.) containing iscove&#39;s modified dulbecco&#39;s medium (IMDM) (Thermo Fisher Scientific) supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine, sodium pyruvate, non-essential amino acids, and where indicated, various concentrations of unconjugated HER2 antibody and Immunoconjugate A of the invention (as prepared according to the examples above). Cells and cell-free supernatants were analyzed after 18 hours via flow cytometry or ELISA. 
     The results of this assay are shown in the figures, for example,  FIG. 1A  (CD40) and  FIG. 1B  (CD86) for Immunoconjugate A on the HCC1954 cell line,  FIG. 1D  (CD40) and  FIG. 1E  (CD86) for Immunoconjugate A on the JIMT-1 cell line, and  FIG. 1G  (CD40) and  FIG. 1H  (CD86) for Immunoconjugate A on the COLO 205 cell line. 
     While the expression of T cell stimulatory molecules such as CD40 and CD86 are necessary for effective T cell activation, APCs also influence the nature of the ensuing immune response through the secretion of proinflammatory cytokines. Therefore, the capacity of immunoconjugates to elicit cytokine secretion in human APCs following stimulation was investigated. The data indicate that the immunoconjugate-stimulated cells secreted high levels of TNFα. See  FIG. 1C  for Immunoconjugate A co-cultured with the HCC1954 cell line,  FIG. 1F  for Immunoconjugate A co-cultured with the JIMT-1 cell line, and  FIG. 1I  for Immunoconjugate A co-cultured with the COLO 205 cell line. 
     Similar data is provided for Immunoconjugates B-I, L-V, K1, and R1 at  FIGS. 2-22D  (without co-culture). 
     Example 143. Assessment of the Pharmacokinetics (PK) Properties of Immunoconjugate P and Immunoconjugate V 
     This example shows that Immunoconjugate P and Immunoconjugate V have favorable PK properties. 
     Cynomolgus primates ( Macaca fascicularis ) were dosed with 10 mg/kg of Immunoconjugate P, Immunoconjugate V, Immunoconjugate W, Immunoconjugate X, Immunoconjugate Y, or Immunoconjugate Z, as shown in Scheme 1, and the PK properties were assessed for 28 days following administration. 
     
       
         
         
             
             
         
       
     
     A trastuzumab PK assay was configured to capture trastuzumab with HCA169 anti-idiotype mAb and to detect with peroxidase labeled HCA176 (HCA176P). An antibody drug conjugate assay was configured to capture trastuzumab with HCA169 anti-idiotype mAb and to detect with a rabbit mAb to A103 followed by detection with peroxidase labeled Goat anti-rabbit IgG. Immunoconjugate P and Immunoconjugate V demonstrated higher serum levels in both PK assays as compared to Immunoconjugate W, Immunoconjugate X, Immunoconjugate Y, and Immunoconjugate Z. 
     All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein. 
     The use of the terms “a” and “an” and “the” and “at least one” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The use of the term “at least one” followed by a list of one or more items (for example, “at least one of A and B”) is to be construed to mean one item selected from the listed items (A or B) or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention. 
     Preferred embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.