Conjugate compounds, methods of making same, and methods and uses thereof

Abstract

A conjugate compound comprises: TTA-X1-MNP-X2-TpA, wherein: TTA is a tumour targeting agent; MNP is a metal nanoparticle; TpA is a therapeutic agent; and X1 and X2 are the same or different, and are each independently selected from a non-covalent linker or a covalent linker. Methods for making the conjugate compound is provided. Methods and uses of the conjugate compound and a composition thereof for treating cancer.

Description

[0001] CONJUGATE COMPOUNDS, METHODS OF MAKING SAME, AND METHODS AND USES THEREOF

[0002] FIELD

[0003] The present disclosure relates generally to conjugate compounds, methods of making same, and methods and uses thereof for treatment of cancer.

[0004] BACKGROUND

[0005] There are many types of cancer treatment such as, and without being limited thereto, biomarker testing for cancer treatment, chemotherapy, hormone therapy, hyperthermia, immunotherapy, photodynamic therapy (PDT), photothermal therapy (PTT), radiation therapy, and others. PTT, for example, is a minimally invasive, local treatment that relies on an optical absorbing agent (i.e. photosensitizer), which can absorb energy and convert it into heat upon stimulating by an electromagnetic radiation (EMR) such as radiofrequency, microwaves, near infrared irradiation, or visible light. PTT is a therapy that induces hyperthermia, including sub-ablative hyperthermia, or ablation in tumour cells. It may be difficult to specifically target cell populations for destruction using PTT. Metal nanoparticles such as gold nanorods (GNRs) can be used as the photosensitizer in PTT.

[0006] During cancer treatment, side-effects are generated from the off-target drug delivery and / or actions of therapeutic agents used in, for example, chemotherapy and immunotherapy.

[0007] There is a need for the development of improved compounds, compositions, uses and / or methods for treatment of cancer.

[0008] The background disclosed herein is included solely to explain the context of the application. This is not to be taken as an admission that any of the material referred to herein was published, known, or part of the common general knowledge as of the priority date.

[0009] SUMMARY

[0010] In an aspect, there is a conjugate compound comprising: TTA-X1-MNP-X2-TpA wherein:

[0011] TTA is a tumour targeting agent;

[0012] MNP is a metal nanoparticle;

[0013] TpA is a therapeutic agent; and

[0014] X1and X2are the same or different, and are each independently selected from a non-covalent linker or a covalent linker. In another aspect, there is a pharmaceutical composition comprising the conjugate compound disclosed herein.

[0015] In another aspect, there is a method for making the compound disclosed herein, the method comprising: combining TTA-CRG1 and CRG2-MNP-X2-TpA to make TTA-X1-MNP- X2-TpA, wherein CRG is a conjugate reactive group.

[0016] In another aspect, there is a method for making the compound disclosed herein, the method comprising: combining TTA-X1-MNP-CRG3 and CRG4-TpA to make TTA-X1-MNP- X2-TpA, wherein CRG3 and CRG4 are different, and are each selected to form X2, optionally, CRG4 is a portion of the TpA, wherein CRG is a conjugate reactive group.

[0017] In another aspect, there is a method for making the compound disclosed herein, the method comprising: combining TTA-CRG1 , CRG2-MNP-CRG3, and CRG4-TpA to make TTA-X1-MNP-X2-TpA, wherein CRG is a conjugate reactive group.

[0018] In another aspect, there is a method for treating cancer, comprising administering to a mammal (e.g. a subject) a therapeutically effective amount of the conjugate compound disclosed herein or the composition disclosed herein.

[0019] In another aspect, there is a use of a therapeutically effective amount of the conjugate compound disclosed herein or the composition disclosed herein for treating cancer.

[0020] It is understood that one or more of the aspects disclosed herein may be combined in any suitable manner. The novel features will become apparent to those of skill in the art upon examination of the following detailed description. It should be understood, however, that the detailed description and the specific examples presented, while indicating certain embodiments of the present invention, are provided for illustration purposes only because various changes and modifications within the spirit and scope of the invention will become apparent to those of skill in the art from the detailed description of the invention and claims that follow.

[0021] BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Reference will now be made, by way of example, to the accompanying drawings and by which the present disclosure can be further understood from the following description with reference to the Figure(s):

[0023] Figures 1A-1 E shows some examples of conjugation for making TTA-X1-GNR- X2- TpA,_wherein: TTA is a tumour targeting agent; GNR is a gold nanorod; TpA is a therapeutic agent; and X1and X2are each DNA linkers.

[0024] Figure 2 shows an example of conjugation using a non-covalent linker to form TTA- GNR. Figure 3 shows an example of conjugation using a covalent linker to form TTA- C(O)NH2-GNR;

[0025] Figure 4 shows an example of conjugation using two complementary ssDNA to form TTA-DNA(1)-GNR.

[0026] Figure 5 shows an example of conjugation using ssDNA that is covalently bound to TpA to form TpA-ssDNA.

[0027] Figure 6 shows an example of conjugation using a portion of a nucleic acid sequence (single strand) of a TpA which is complementary to a ssDNA and forms a H-S- DNA-TpA conjugate having a thiol to bind to GNRs.

[0028] Figure 7 shows an example of conjugation of TTA-X1-GNR with ssDNA to form TTA-X1-GNR-ssDNA.

[0029] Figure 8 shows an example of conjugation of TTA-X1-GNR-ssDNA with ssDNA-TpA to form TTA-X1-GNR-DNA-TpA.

[0030] Figure 9 shows an example of conjugation of TTA-X1-GNR with H-S-DNA-TpA to form TTA-X1-GNR-DNA-TpA.

[0031] Figure 10 shows an example of a lateral flow assay stick, made from nitro cellulose sheets, biomolecule of choice (at 1 mg / mL in PBS, of 0.5 uL spot volume) spotted to create control spot and anti-species antibodies used (1 mg / mL in PBS, 0.5 uL spot volume) to create test spots. Also, depicting a relative position of the test and control spots in relation to conjugate pad.

[0032] Figure 11 shows a scheme for conjugation of Fc Tag of IL2 / IL15 using SATA reagent and GNRs.

[0033] Figure 12 shows UV-VIS Scans of various GNR solutions.

[0034] Figure 13a and 13b shows an example of IL2 developed Lateral Flow Assay (LFA) test sticks. Figure 13a shows binding only through anti-species (control) and Figure 13b shows binding through both anti-species and anti-l L2 Ab (control)

[0035] Figures 14a and 14b: Figure 14a shows no binding of GNR-PD1 conjugate on a LFA tests made from PSMA protein (test spot) and anti-PSMA Ab (control spot) and Figure 14b shows a polyclonal Goat anti-human (GAH) antibody immobilized on a nitrocellulose membrane as control spot and human CD47 (aa 18-135) recombinant protein was immobilized as a test spot.

[0036] Figure 15 shows a scheme for thiol-functionalization of PSMA Binding Motif (PSMABM) (SATA Chemistry) to make it ready for gold-sulfur bonds with GNRs.

[0037] Figure 16 shows UV-VIS Scans with a change in position of peak between 800- 900nm (LSPR, longitudinal surface plasmon resonance) for starting GNRs and PSMABM- GNR conjugates. Figure 17 shows an example of IL2 and PSMA LFA stick spotted with anti-ILs and GAH, and PSMA protein and Anti PSMA antibodies. PSMABM with both types of GNRs, show successful binding to PSMA protein.

[0038] Figure 18 shows a scheme for SATA modification of anti-human CD47.

[0039] Figures 19a and 19b: Figure 19a shows LFA tests that show binding of human CD47 recombinant protein with GNR-CD47 conjugate and Figure 19b shows no binding of GNR-CD47 conjugate on IL2 stick.

[0040] Figure 20 shows a scheme for the synthesis of PSMABM-GNR-IL2 from either GNR-IL2 or PSMABM-GNR conjugates using HS-PSMABM or HS-IL2, respectively.

[0041] Figures 21a and 21 b: Figure 21a shows binding of PSMABM-GNR-IL2 conjugate to test spot (PSMA protein) and Figure 21 b shows the binding of PSMABM-GNR-IL2 to test spot (anti-l L2 antibody) and control spot (GAH) to demonstrate formation of PSMABM- GNR-IL2 conjugate compound.

[0042] Figure 22 shows UV-Vis traces comparing washed GNRs, half-conjugate and conjugate compounds for PSMABM-GNR-IL2. LSPR values: GNR = 842 nm; PSMABM- GNR = 842 nm; PSMABM-GNR-IL2 = 856 nm.

[0043] Figure 23 shows a scheme for the synthesis of PSMABM-GNR-PD1 from either GNR-PD1 or PSMABM-GNR conjugates using HS-PSMABM or HS-PD1 , respectively.

[0044] Figure 24 shows LFA testing of PSMABM-GNR-PD1 and binding only to PSMA protein. The binding of PSMABM-SH to GNR-PD1 conjugate (option 2 of Figure 23) was confirmed via the development of a test spot for the LFA stick spotted with PSMA protein.

[0045] Figure 25 shows UV-Vis traces comparing washed GNRs, half-conjugate and conjugate compound for PSMABM-GNR-PD1 . LSPR values: GNR = 842 nm; GNR-PD1 = 842 nm; PSMABM-GNR-PD1 = 852 nm.

[0046] Figure 26 shows a scheme for the synthesis of CD47-GNR-IL2 from either GNR-IL2 or CD47-GNR half-conjugates using CD47-SH or HS-IL2, respectively.

[0047] Figures 27a and 27b shows confirmed formation of CD47-GNR-IL2 conjugate throughthe development of a test spot for the LFA stick spotted with CD47 protein (Figure 27a) and throughthe development of a test and control spot for the LFA stick spotted with anti- IL2 Abs (Figure 27b) and GAH Abs control.

[0048] Figure 28 shows UV-Vis traces comparing washed GNRs, half-conjugate and conjugate compound for CD47-GNR-IL2. LSPR values: GNR = 840 nm; CD47-GNR = 838 nm; CD47-GNR-IL2 = 850 nm.

[0049] Figure 29 shows a scheme for the synthesis of CD47-GNR-PD1 from either GNR- PD1 or CD47-GNR half-conjugates using CD47-SH or HS-PD1 , respectively. Figure 30 shows LFA testing of CD47-GNR-PD1 on a CD47 LFA stick. The binding of CD47-SH to GNR-PD1 conjugate (option 2 of Figure 29) was confirmed via the development of a test spot for the LFA stick spotted with CD47 protein.

[0050] Figure 31 shows UV-Vis traces comparing washed GNRs, half-conjugate conjugate compound for CD47-GNR-PD1 . LSPR values: GNR = 842 nm; CD47-GNR = 842 nm; CD47-GNR-PD1 = 840 nm.

[0051] Figure 32 shows a scheme for dsDNA modification of PD1 lgG4 antibody.

[0052] Figures 33a to 33c: Figure 33a shows no binding of GNR-dsDNA-PD1 to LFA stick spotted with PSMA protein, similarly Figure 33b shows no binding of GNR-dsDNA-PD1 with to LFA stick spotted with CD47. Figure 33c shows the binding between GNR-dsDNA- PD1 conjugate and polyclonal Goat anti-human (GAH) antibody immobilized on nitrocellulose membrane to show the formation of GNR-dsDNA-PD1 .

[0053] Figure 34 shows a scheme for dsDNA modification of IL2-Fc.

[0054] Figures 35a and 35b: Figure 35a shows no binding of GNR-dsDNA-IL2 to LFA stick spotted with PSMA protein and shows binding to both test and control spots for the LFA stick spotted with anti-l L2 Abs (test) and GAH Abs (control) (Figure 35b).

[0055] Figure 36 shows UV-Vis traces comparing GNR to GNR-dsDNA-IL2. LSPR values: GNR = 842 nm; GNR-ds-DNA-IL2 = 850 nm.

[0056] Figure 37 shows a scheme for making CD47-GNR-dsDNA-PD1 conjugate.

[0057] Figure 38 shows LFA testing of CD47-GNR-dsDNA-PD1 on a CD47 LFA stick. The binding of CD47-SH to GNR-dsDNA-PD1 conjugate (option 1 of Figure 37) was confirmed via the development of a test spot for the LFA stick spotted with CD47 protein.

[0058] Figure 39 shows UV-Vis traces comparing washed GNRs and labile conjugate compound for CD47-GNR-dsDNA-PD1. LSPR values: GNR = 842 nm; CD47-GNR- dsDNA-PD1 = 848 nm.

[0059] Figure 40 shows a protocol described in Hermanson, G.T., “Bioconjugation Techniques”, 3rdedition, page 886.

[0060] DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS / ASPECTS

[0061] Definitions

[0062] Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those disclosed herein can be used in the practice for testing of the present invention, the typical materials and methods are disclosed herein. In addition, in describing and claiming the present invention, the common terminology generally used is disclosed herein below. If a term is used in this disclosure but is not specifically disclosed herein, the definition from the IUPAC Compendium of Chemical Terminology can be applied, as long as that definition does not conflict with any other disclosure or description applied herein, or render indefinite or non-enabled any claim to which that definition is applied. All references herein to elements or metals belonging to a certain Group refer to the Periodic Table of the Elements and Hawley's Condensed Chemical Dictionary, 13th Edition. Also, any references to the Group or Groups shall be to the Group or Groups as reflected in the Periodic Table of Elements using the CAS system for numbering groups. To the extent that any definition, description or usage provided by any document incorporated herein by reference conflicts with the description or usage provided herein, the description or usage provided herein controls. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting.

[0063] The term “metal nanoparticle” or “MNP” can refer to an average particle size that is less than about 1 m in size, and typically from about 1 nm to about 900 nm in size and various ranges therebetween. An MNP can take a variety of shapes, including but not limited to, tube, rod, shell, cage, sphere, fiber, wire, stars, plate, sea archon, or a combination thereof.

[0064] The term “therapeutic agent” or “TpA” can refer to any substance having a therapeutic property that can produce a desired, usually beneficial, effect. For example, TpAs can be any agent for treating a symptom or disease in an individual in need of such treatment. TpAs may treat, ameliorate, and / or prevent disease. TpAs, as disclosed herein, may be biologies or synthetics, small or macromolecule therapeutics, or combinations thereof. TpA may be referred to a payload.

[0065] The term “chemotherapeutic agent” as used herein refers to any therapeutic agent used to treat cancer.

[0066] The term “tumour targeting agent” or “TTA” can refer to any suitable substance that is capable of specifically binding to or recognizing a target cell such as, and without being limited thereto, cancer cells (e.g. tumour cells) for the targeted delivery of conjugate compound or the payload. TTAs can include all substances capable of binding to cancer cells (e.g. the tumour), for example, the tumour microenvironment (e.g. tumour vasculature and stroma). In the case of a solid tumor, the TTA may take advantage of an EPR (enhanced permeability and retention) effect to ensure high concentration of the conjugate compound in the tumor compared to the rest of the body or surrounding heathy tissue (Z. Fu et al., Antibody drug conjugate: the “biological missile” for targeted cancer therapy, Signal Transduction and Targeted Therapy (2022)7:93, 1-25). The term “aptamer” can refer to a class of nucleic acids that are composed of RNA or DNA oligonucleotides that have high specificity and affinity for their associated targets and may offer unique chemical and biological characteristics in terms of biochemical activity, molecular recognition or binding attributes. Usually, and for use herein, the aptamer has a molecular activity such as binding to a target molecule at a specific epitope (region).

[0067] The terms “cancer”, “tumour” and “carcinoma” refer to cells that exhibit relatively abnormal, uncontrolled, and / or autonomous growth, so, for example, they exhibit an aberrant growth phenotype characterized by a significant loss of control of cell proliferation. The term “cancer” is understood to be broad and encompass, for example, “tumour” and “carcinoma”.

[0068] "Variants" of the sequences disclosed herein are biologically active sequences that have a peptide sequence that differs from the sequence of a native or wild-type sequence, by virtue of an insertion, deletion, modification and / or substitution of one or more amino acids within the native sequence. Such variants generally have less than 100% sequence identity with a native sequence. Ordinarily, however, a biologically active variant will have an amino acid sequence with at least about 70% sequence identity with the sequence of a corresponding naturally occurring sequence, typically at least about 75%, more typically at least about 80%, even more typically at least about 85%, even more typically at least about 90%, and even more typically of at least about 95%, 96%, 97%, 98%, or 99% sequence identity. The variants nucleotide fragments of any length that retain a biological activity of the corresponding native sequence. Variants also include sequences wherein one or more amino acids are added at either end of, or within, a native sequence. Variants also include sequences where a number of amino acids are deleted and optionally substituted by one or more different amino acids.

[0069] "Percent sequence identity" is defined herein as the percentage of amino acid residues in the candidate sequence that are identical with the residues in the sequence of interest after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. None of 5', 3', or internal extensions, deletions or insertions into the candidate sequence shall be construed as affecting sequence identity or homology. Methods and computer programs for the alignment are well known in the art, such as "BLAST".

[0070] An “immunomodulator” refers to any modulator that can provide (e.g. change, start, stop, increase, decrease, etc.) an immune response (e.g. molecule, heat, radiation, etc.).

[0071] The term "polynucleotide" as used herein is defined as a chain of nucleotides. Furthermore, nucleic acids are polymers of nucleotides. Thus, nucleic acids and polynucleotides as used herein are interchangeable. One skilled in the art has the general knowledge that nucleic acids are polynucleotides, which can be hydrolyzed into the monomeric "nucleotides." The monomeric nucleotides can be hydrolyzed into nucleosides. As used herein polynucleotides include, but are not limited to, all nucleic acid sequences which are obtained by any means available in the art, including, without limitation, recombinant means, i.e., the cloning of nucleic acid sequences from a recombinant library or a cell genome, using ordinary cloning technology and PCR, and the like, and by synthetic means. The terms “nucleic acid,” “nucleic acid molecule,” “nucleic acid sequence,” and “polynucleotide” are used interchangeably and are intended to include, but are not limited to, a polymeric form of nucleotides covalently linked together that may have various lengths, either deoxyribonucleotides or ribonucleotides, or analogs, derivatives or modifications thereof. Different polynucleotides may have different three-dimensional structures, and may perform various functions, known or unknown. Non-limiting examples of polynucleotides include a gene, a gene fragment, an exon, an intron, intergenic DNA (including, without limitation, heterochromatic DNA), messenger RNA (mRNA), transfer RNA, ribosomal RNA, a ribozyme, cDNA, a recombinant polynucleotide, a branched polynucleotide, a plasmid, a vector, isolated DNA of a sequence, isolated RNA of a sequence, a nucleic acid probe, and a primer. Polynucleotides useful may include natural nucleic acid sequences and variants thereof, artificial nucleic acid sequences, or a combination of such sequences.

[0072] The terms “nucleic acid oligomer” and “oligonucleotide” are used interchangeably and are intended to include, but are not limited to, nucleic acids having a length of 200 nucleotides or less. In some embodiments, an oligonucleotide is a nucleic acid having a length of 2 to 200 nucleotides, 2 to 150 nucleotides, 5 to 150 nucleotides or 5 to 100 nucleotides. The terms “polynucleotide,” “oligonucleotide,” “oligo” or the like refer, in the usual and customary sense, to a linear sequence of nucleotides. Oligonucleotides are typically from about 5, 6, 7, 8, 9, 10, 12, 15, 25, 30, 40, 50 or more nucleotides in length, up to about 100 nucleotides in length. In some embodiments, an oligonucleotide is a primer configured for extension by a polymerase when the primer is annealed completely or partially to a complementary nucleic acid template. A primer is often a single stranded nucleic acid. In certain embodiments, a primer, or portion thereof, is substantially complementary to a portion of an adapter. In some embodiments, a primer has a length of 200 nucleotides or less. In certain embodiments, a primer has a length of 10 to 150 nucleotides, 15 to 150 nucleotides, 5 to 100 nucleotides, 5 to 50 nucleotides or 10 to 50 nucleotides.

[0073] As used herein, the terms "peptide," "polypeptide," and "protein" are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein's or peptide's sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which can be referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. "Polypeptides" include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof.

[0074] The term "antibody" ("Ab"), “antibodies” (“Abs”), or "immunoglobulin" (Ig) refers to any form of a peptide, polypeptide derived from, modeled after or encoded by, an immunoglobulin gene, or fragment thereof, that is capable of binding an antigen (Ag) or epitope. See, e.g., Immunobiology, Fifth Edition, C. A. Janeway, P. Travers, M., Walport, M. J. Shlomchiked., ed. Garland Publishing (2001). The term "antibody" is used herein in the broadest sense, and encompasses monoclonal, polyclonal or multispecific antibodies, multivalent antibodies, minibodies, heteroconjugates, diabodies, triabodies, antibody mimics, chimeric antibodies, synthetic antibodies, humanized antibodies, antibody fragments, and binding agents that employ the complementarity determining regions (CDRs) (or variants thereof that retain antigen binding activity) of the parent antibody. Antibodies are defined herein as retaining at least one desired activity (i.e. desired level of binding activity for the antigen) of the parent antibody. It also includes functional equivalents of the antibodies of the presently disclosed subject matter. The term “functional equivalent” regarding an antibody, refers to a molecule that has binding characteristics that are comparable to those of a given antibody. In some embodiments, chimerized, humanized, antibody variants, and single chain antibodies, as well as fragments thereof, are considered functional equivalents of the corresponding antibodies upon which they are based.

[0075] The term "antibody fragment" can refer to a portion of an intact antibody that includes the antigen binding site or variable domains of an intact antibody, wherein the portion can be free of the constant heavy chain domains (e.g., CH2, CH3, and CH4) of the Fc region of the intact antibody. Alternatively, portions of the constant heavy chain domains (e.g., CH2, CH3, and CH4) can be included in the "antibody fragment". Antibody fragments retain antigen-binding and include Fab, Fab', F(ab')2, Fc, and Fv fragments; diabodies; triabodies; single-chain antibody molecules (sc-Fv); minibodies, nanobodies, and multispecific antibodies formed from antibody fragments. Papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment, whose name reflects its ability to crystallize readily. Pepsin treatment yields an F(ab')2 fragment that has two antigencombining sites and is still capable of binding to an antigen. By way of example, a Fab fragment also contains the constant domain of a light chain and the first constant domain (CH1) of a heavy chain. "Fv" is the minimum antibody fragment that contains a complete antigen-recognition and -binding site. This region consists of a dimer of one heavy chain and one light chain variable domain in tight, non-covalent association. It is in this configuration that the three hypervariable regions of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six hypervariable regions confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three hypervariable regions specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site. "Single-chain Fv" or "sFv" antibody fragments comprise the VH and VL domains of antibody, wherein these domains are present in a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the sFv to form the desired structure for antigen binding. For a review of sFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

[0076] The term "diabodies" refers to small antibody fragments with two antigen-binding sites, which fragments comprise a heavy chain variable domain (VH) connected to a light chain variable domain (VL) in the same polypeptide chain (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigenbinding sites. Diabodies are described more fully in, for example, EP 404,097; WO 93 / 11161 ; and Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993).

[0077] The term "specific" or "specificity" in the context of antibody-antigen interactions refers to the selective, non-random interaction between an antibody and its target epitope. Here, the term "antigen" refers to any suitable molecule that can be, for example, a biomolecule, small or macromolecule, or combinations of both, a cell receptor, or checkpoint on cell surface that is recognizable with antibody, aptamer etc. The specific portion of an antigen that is bound by an antibody is termed the "epitope". This interaction depends on the presence of structural, hydrophobic / hydrophilic, and / or electrostatic features that allow appropriate chemical or molecular interactions between the molecules. Thus, an antibody is commonly said to "bind" (or "specifically bind") or "reactive with" (or "specifically reactive with), or, equivalently, "reactive against" (or "specifically reactive against") the epitope of its target antigen. Antibodies are commonly described in the art as being "against" or "to" their antigens as shorthand for antibody binding to the antigen. Thus an "antibody that binds C1 P," an "antibody reactive against C1 P," an "antibody reactive with C1 P," an "antibody to C1 P" and an "anti-C1 P antibody" all have the same meaning in the art. Antibody molecules can be tested for specificity of binding by comparing binding to the desired antigen to binding to unrelated antigen or analogue antigen or antigen mixture under a given set of conditions. Preferably, an antibody will lack significant binding to unrelated antigens, or even analogs of the target antigen.

[0078] As used herein, the terms “bind” and “bound” can refer to an association between atoms or molecules. The association can be direct or indirect. For example, bound atoms or molecules may be directly bound to one another, e.g., by a covalent bond or non- covalent bond (e.g. electrostatic interactions (e.g. ionic bond, hydrogen bond, halogen bond), van der Waals interactions (e.g., dipole-dipole, dipole-induced dipole, London dispersion), ring stacking (pi effects), hydrophobic interactions and the like). As a further example, two molecules may be bound indirectly to one another by way of direct binding to one or more intermediate molecules, thereby forming a complex.

[0079] The term “conjugate” as use herein refers to a molecule (e.g. biomolecule) comprising at least two moieties. For example, and without limitation, the moieties can be connected via a linker (e.g. covalent and / or non-covalent linker).

[0080] The term “non-covalent linker” can refer to a non-covalent bond (e.g., electrostatic interactions (e.g., ionic bond, hydrogen bond, etc.) or van der Waals interactions (e.g., dipole-dipole, dipole-induced dipole, London dispersion, etc.) between two moieties / molecules. For example, the non-covalent linker is the result of two molecules (e.g. TTA and the MNP) that are not covalently linked to each other and interact with each other via a non-covalent bond.

[0081] The term “covalent linker” can refer to a divalent moiety, which connects at least two molecules (e.g. TTA and the MNP).

[0082] The term “cross-linker” refers to a molecule comprising a moiety that can be crosslinked. For example, a cross-linker can refer to a moiety that links one entity to another entity. In some embodiments, linkage (i.e., the “cross-link”) between two entities is or comprises a covalent bond. In some embodiments, linkage between two entities is or comprises an ionic bond. In some embodiments, a cross-linker comprises a photo-sensitive functional group. In some embodiments, a cross-linker comprises a pH-sensitive functional group. In some embodiments, a cross-linker comprises a thermal-sensitive functional group. The cross-linker can be any suitable molecule capable of cross-linking. The molecule can be any suitable compound , such as but not limited to, a substituted or unsubstituted hydrocarbon group, a substituted or unsubstituted heterogeneous group, a substituted or unsubstituted carbocyclic group, a substituted or unsubstituted heterocyclic group, substituted or unsubstituted aromatic group, or a substituted or unsubstituted heteroaromatic group. Further examples include a substituted or unsubstituted hydrocarbon group, a chain of ethoxy groups, substituted or unsubstituted phenyl groups, etc. or combination thereof.

[0083] The term “conjugate reactive moiety” and “conjugate reactive group” (CRG) refers to a) reactive group for forming a covalent linker as a result of the association between atoms or molecules of each CRG , which can include a linking functional group; b) one or more CRGs can include a cross-linker; or c) forming a non-covalent linker as a result of the association between atoms or molecules. The association can be direct or indirect. The CRGs can be linked through their reactive groups (e.g. zero length linker or any suitable length linker). A further linker can be added separately, linking the CRGs, such as a third bifunctional molecule through establishment of chemical bonds. Bi-functional molecule can be a homo-functional with both functional groups being the same or a hetero-functional whereby the two functional groups are different from one another.

[0084] The term “click chemistry” refers to a term that was introduced by K. B. Sharpless in 2001 to describe reactions that may be, for example, high yielding, wide in scope, create by-products that can be removed without chromatography, may be stereospecific, may be simple to perform, and may be conducted in easily removable or inert solvents. Several types of reaction have been identified that fulfill these criteria, thermodynamically-favored reactions that lead specifically to one product, such as nucleophilic ring opening reactions of epoxides and aziridines, non-aldol type carbonyl reactions, such as formation of hydrazones and heterocycles, additions to carboncarbon multiple bonds, such as oxidative formation of epoxides and Michael additions, and cycloaddition reactions.

[0085] The term “linking functional” group of a covalent linker, which is a result of the association between atoms or molecules of each conjugate reactive group.

[0086] The term “cleavable linker” is used in accordance with its ordinary meaning and refers to a covalent linker or a non-covalent linker which is capable of being separated (e.g., detached, split, broken bond, disconnected, hydrolyzed, a stable bond within the linker is broken) into distinct entities. A cleavable linker may be cleavable in response to stimulus (e.g., photo-irradiation such as irradiation at certain wavelength(s) to control cleavage, for example and without being limited thereto, UV, visible, or NIR), enzymes, nucleophilic / basic reagents, reducing agents, electrophilic / acidic reagents, organometallic and metal reagents, or oxidizing reagents). The stimulus may be external or internal. The terms “single strand” and “ssDNA” are used in accordance with its plain and ordinary meaning and refer to a single-stranded polynucleotide. It may be a portion of a gene, a regulatory sequence, genomic DNA, cDNA, RNA including mRNA, miRNA, rRNA, or others.

[0087] The term “polymer” refers to macromolecules having one or more structurally unique repeating units. The repeating units are referred to as “monomers,” which are polymerized for the polymer. Typically, a polymer is formed by monomers linked in a chainlike structure. A polymer formed entirely from a single type of monomer is referred to as a “homopolymer.” A polymer formed from two or more unique repeating structural units may be referred to as a “copolymer.” A polymer may be linear or branched, and may be random, block, polymer brush, hyperbranched polymer, bottlebrush polymer, dendritic polymer, or polymer micelles. The term “polymer” includes homopolymers, copolymers, tripolymers, tetra polymers and other polymeric molecules made from monomeric subunits. Copolymers include alternating copolymers, periodic copolymers, statistical copolymers, random copolymers, block copolymers, linear copolymers and branched copolymers. The term “polymerizable monomer” is used in accordance with its meaning in the art of polymer chemistry and refers to a compound that may covalently bind chemically to other monomer molecules (such as other polymerizable monomers that are the same or different) to form a polymer. Polymers can be hydrophilic, hydrophobic, or amphiphilic, as known in the art. Thus, “hydrophilic polymers” are substantially miscible with water and include, but are not limited to, polyethylene glycol and the like. “Hydrophobic polymers” are substantially immiscible with water and include, but are not limited to, polyethylene, polypropylene, polybutadiene, polystyrene, polymers disclosed herein, and the like. “Amphiphilic polymers” have both hydrophilic and hydrophobic properties and are typically copolymers having hydrophilic segment(s) and hydrophobic segment(s). Polymers include homopolymers, random copolymers, and block copolymers, as known in the art.

[0088] The term “homopolymer” can refer to a polymer having a single monomeric unit. The term “copolymer” refers to a polymer derived from two or more monomeric species. The term “random copolymer” refers to a polymer derived from two or more monomeric species with no preferred ordering of the monomeric species. The term “block copolymer” refers to polymers having two or homopolymer subunits linked by covalent bond. Thus, the term “hydrophobic homopolymer” refers to a homopolymer which is hydrophobic. The term “hydrophobic block copolymer” refers to two or more homopolymer subunits linked by covalent bonds and which is hydrophobic.

[0089] As used herein, the term “complementary” or “substantially complementary” refers to the hybridization, base pairing, or the formation of a duplex between nucleotides or nucleic acids. For example, complementarity exists between the two strands of a double- stranded DNA molecule or between an oligonucleotide primer and a primer binding site on a single-stranded nucleic acid when a nucleotide (e.g., RNA or DNA) or a sequence of nucleotides is capable of base pairing with a respective cognate nucleotide or cognate sequence of nucleotides. When referring to a double-stranded polynucleotide including a first strand hybridized to a second strand, it is to be understood that each of the terms “first strand” and “second strand” refer to single-stranded polynucleotides. As disclosed herein and commonly known in the art the complementary (matching) nucleotide of adenosine (A) is thymidine (T) or uracil (U) and the complementary (matching) nucleotide of guanosine (G) is cytosine (C). Thus, a complement may include a sequence of nucleotides that base pair with corresponding complementary nucleotides of a second nucleic acid sequence. The nucleotides of a complement may partially or completely match the nucleotides of the second nucleic acid sequence. Where the nucleotides of the complement completely match each nucleotide of the second nucleic acid sequence, the complement forms base pairs with each nucleotide of the second nucleic acid sequence. Where the nucleotides of the complement partially match the nucleotides of the second nucleic acid sequence only some of the nucleotides of the complement form base pairs with nucleotides of the second nucleic acid sequence. Examples of complementary sequences include coding and noncoding sequences, wherein the non-coding sequence contains complementary nucleotides to the coding sequence and thus forms the complement of the coding sequence. A further example of complementary sequences are sense and antisense sequences, wherein the sense sequence contains complementary nucleotides to the antisense sequence and thus forms the complement of the antisense sequence. “Duplex” means at least two oligonucleotides and / or polynucleotides that are fully or partially complementary undergo Watson-Crick type base pairing among all or most of their nucleotides so that a stable complex is formed. Complementary single stranded nucleic acids and / or substantially complementary single stranded nucleic acids can hybridize to each other under hybridization conditions, thereby forming a nucleic acid that is partially or fully double stranded. When referring to a double-stranded polynucleotide including a first strand hybridized to a second strand, it is understood that each of the first strand and the second strand are independently single-stranded polynucleotides. All or a portion of a nucleic acid sequence may be substantially complementary to another nucleic acid sequence, in some embodiments. As referred to herein, “substantially complementary” refers to nucleotide sequences that can hybridize with each other under suitable hybridization conditions. Hybridization conditions can be altered to tolerate varying amounts of sequence mismatch within complementary nucleic acids that are substantially complementary. Substantially complementary portions of nucleic acids that can hybridize to each other can be 75% or more, 76% or more, 77% or more, 78% or more, 79% or more, 80% or more, 81% or more, 82% or more, 83% or more, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more or 99% or more complementary to each other. In some embodiments substantially complementary portions of nucleic acids that can hybridize to each other are 100% complementary. Nucleic acids, or portions thereof, that are configured to hybridize to each other often include nucleic acid sequences that are substantially complementary to each other.

[0090] As disclosed herein, the complementarity of sequences may be partial, in which only some of the nucleic acids match according to base pairing, or complete, where all the nucleic acids match according to base pairing. Thus, two sequences that are complementary to each other, may have a specified percentage of nucleotides that complement one another (e.g., about 60%, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher complementarity over a specified region). In embodiments, two sequences are complementary when they are completely complementary, having 100% complementarity. In embodiments, sequences in a pair of complementary sequences form portions of a single polynucleotide with non-base- pairing nucleotides (e.g., as in a hairpin or loop structure, with or without an overhang) or portions of separate polynucleotides. In embodiments, one or both sequences in a pair of complementary sequences form portions of longer polynucleotides, which may or may not include additional regions of complementarity.

[0091] As used herein, "treatment", “treating”, or “therapy” is an approach for obtaining beneficial or desired clinical results. For the purposes disclosed herein, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" and “therapy” can also mean prolonging survival as compared to expected survival if not receiving treatment or therapy. Thus, "treatment" or “therapy” is an intervention performed with the intention of altering the pathology of a disorder. Specifically, the treatment or therapy may directly prevent, slow down or otherwise decrease the pathology of a disease or disorder such as cancer, or may render the cells more susceptible to treatment or therapy by other therapeutic agents.

[0092] The terms "therapeutically effective amount", "effective amount" or "sufficient amount" mean a quantity sufficient, when administered to a subject, including a mammal, for example a human, to achieve a desired result, for example an amount effective to treat cancer. Effective amounts of the metal nanorods disclosed herein may vary according to factors such as the disease state, age, sex, and weight of the subject. Dosage or treatment regimes may be adjusted to provide the optimum therapeutic response, as is understood by a skilled person.

[0093] Moreover, a treatment regime of a subject with a therapeutically effective amount may consist of a single administration, or alternatively comprise a series of applications. The length of the treatment period depends on a variety of factors, such as the severity of the disease, the age of the subject, the concentration of the agent / compound / medicament, the responsiveness of the patient to the agent / compound / medicament compound / medicament, or a combination thereof. It will also be appreciated that the effective dosage of the agent / compound / medicament used for the treatment may increase or decrease over the course of a particular treatment regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. The conjugate compounds disclosed herein may, in embodiments, be administered before, during or after treatment with conventional therapies for the disease or disorder in question, such as cancer.

[0094] The terms “diagnostic effective amount” or “imaging effective amount” mean a quantity sufficient, when administered to a subject, including a mammal, for example a human, to achieve a desired result, for example an amount effective to diagnose or image a tumour. In embodiments, a diagnostic or imaging effective amount is distinct from a therapeutically effective amount.

[0095] The term "subject" as used herein refers to any member of the animal kingdom, including birds, fish, invertebrates, amphibians, mammals, and reptiles. Typically, the subject is a human or non-human vertebrate. Non-human vertebrates include livestock animals, companion animals, and laboratory animals. Non-human subjects also specifically include non-human primates as well as rodents. Non-human subjects also specifically include, without limitation, poultry, chickens, horses, cows, pigs, goats, dogs, cats, guinea pigs, hamsters, mink, rabbits, crustaceans, and molluscs. Typically, the subject is a mammal. The term "mammal" refers to any animal classified as a mammal, including humans, other higher primates, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, cats, cattle, horses, sheep, pigs, goats, rabbits, etc. Typically, the mammal is human.

[0096] The term “pharmaceutically acceptable” means that the compound or combination of compounds is compatible with the remaining ingredients of a formulation for pharmaceutical use, and that it is generally safe for administering to humans according to established governmental standards, including those promulgated by the United States Food and Drug Administration.

[0097] The term "pharmaceutically acceptable carrier" includes, but is not limited to solvents, dispersion media, coatings, antibacterial agents, antifungal agents, isotonic and / or absorption delaying agents and the like. The use of pharmaceutically acceptable carriers is well known.

[0098] The term “non-toxic” refers to the non-occurrence of pathological phenomena as a result of using pharmacological levels of the metal nanorods disclosed herein. The term substantially non-toxic is defined as including acceptably low toxicity as well as nontoxicity.

[0099] "Substantially free" of an element / feature herein means less than about 5%, typically less than about 2%, more typically less than about 1%, even more typically less than about 0.5%, most typically less than about 0.1% of the element / feature.

[0100] A “mixture” or “combination” are terms that may be used interchangeably. A mixture is not limited to two or more components that have been mixed. A mixture may be two or more components combined without having been mixed.

[0101] The terms inhibit, reduced, prevented, minimized, or delayed, can be used interchangeably. These terms can refer to partially, substantially, or completely slowing, hindering, reducing, delaying, or preventing.

[0102] The term "hydrocarbon” group or “hydrocarbyl” group means a chain of carbon atoms, typically 10 to 25 carbon atoms and more typically 12 to 24 carbon atoms. Hydrocarbon groups may have a linear or branched chain structure. Typical hydrocarbon groups have one or two branches, typically one branch. Typically, hydrocarbon groups are saturated. Unsaturated hydrocarbon groups may have one or more double bonds, one or more triple bonds, or combinations thereof. Typical unsaturated hydrocarbon groups have one or two double bonds or one triple bond; more typically unsaturated hydrocarbon groups have one double bond.

[0103] The term "heterogeneous” group means a saturated or unsaturated chain of nonhydrogen member atoms comprising carbon atoms and at least one heteroatom. Heterogeneous groups typically have 1 to 25 member atoms. More typically, the chain contains 1 to 12 member atoms, 1 to 10, and most typically 1 to 6. The chain may be linear or branched. Typical branched heterogeneous groups have one or two branches, more typically one branch. Typically, heterogeneous groups are saturated. Unsaturated heterogeneous groups may have one or more double bonds, one or more triple bonds, or both. Typical unsaturated heterogeneous groups have one or two double bonds or one triple bond. More typically, the unsaturated heterogeneous group has one double bond.

[0104] When the term "unsaturated" is used in conjunction with any group, the group may be fully unsaturated or partially unsaturated. However, when the term “unsaturated” is used in conjunction with a specific group defined herein, the term maintains the limitations of that specific group. Where the term "alkyl” group is used, either alone or within other terms such as "haloalkyl” group and "alkylamino” group, it encompasses linear or branched carbon radicals having, for example, one to about twenty-five carbon atoms (depending on whether or not hydrophobicity is required) or, in specific embodiments, one to about twelve carbon atoms. In other embodiments, alkyl groups are "lower alkyl" groups having one to about six carbon atoms. Examples of such groups include, but are not limited thereto, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl and the like. In more specific embodiments, lower alkyl groups have one to four carbon atoms. Other alkyl groups encompass linear or branched carbon radicals having, for example, twelve to about twenty-five carbon atoms. Typically, alkyl groups are saturated. Unsaturated alkyl groups may have one or more double bonds, one or more triple bonds, or combinations thereof. Typical unsaturated alkyl groups have one or two double bonds or one triple bond; more typically unsaturated alkyl groups have one double bond.

[0105] The term "halo" means halogens such as fluorine, chlorine, bromine or iodine atoms.

[0106] The term "haloalkyl” group encompasses groups wherein any one or more of the alkyl carbon atoms is substituted with halo as defined above. Specifically encompassed are monohaloalkyl, dihaloalkyl and polyhaloalkyl groups including perhaloalkyl. A monohaloalkyl group, for one example, may have either an iodo, bromo, chloro or fluoro atom within the group. Dihalo and polyhaloalkyl groups may have two or more of the same halo atoms or a combination of different halo groups. "Lower haloalkyl” group encompasses groups having 1- 6 carbon atoms. In some embodiments, lower haloalkyl groups have one to three carbon atoms. Examples of haloalkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl, pentafluoroethyl, heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl and dichloropropyl.

[0107] The term "hydroxy a Iky I” group encompasses linear or branched alkyl groups having, for example and without being limited thereto, one to about ten carbon atoms, any one of which may be substituted with one or more hydroxyl groups. In embodiments, hydroxyalkyl groups are "lower hydroxyalkyl" groups having one to six carbon atoms and one or more hydroxyl groups. Examples of such groups include hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl and hydroxyhexyl.

[0108] The term "alkoxy” group or “alkoxyalkyl” group encompasses linear or branched oxy- containing groups each having alkyl portions of, for example and without being limited thereto, one to about ten carbon atoms. In embodiments, alkoxy groups are "lower alkoxy" groups having one to six carbon atoms. Examples of such groups include methoxy, ethoxy, propoxy, butoxy and tert-butoxy. In certain embodiments, lower alkoxy groups have one to three carbon atoms. The "alkoxy" groups may be further substituted with one or more halo atoms, such as fluoro, chloro or bromo, to provide "haloalkoxy" groups. In other embodiments, lower haloalkoxy groups have one to three carbon atoms. Examples of such groups include fluoromethoxy, chloromethoxy, trifluoromethoxy, trifluoroethoxy, fluoroethoxy, and fluoropropoxy.

[0109] The term "aromatic” group or “aryl” group means an aromatic group having one or more rings wherein such rings may be attached together in a pendent manner or may be fused. In particular embodiments, an aromatic group is one, two or three rings. Monocyclic aromatic groups may contain 4 to 10 carbon atoms, typically 4 to 7 carbon atoms, and more typically 4 to 6 carbon atoms in the ring. Typical polycyclic aromatic groups have two or three rings. Polycyclic aromatic groups have two rings typically having 8 to 12 carbon atoms, typically 8 to 10 carbon atoms in the rings. Examples of aromatic groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl.

[0110] The term "heteroaromatic” group or “heteroaryl” group means an aromatic group having one or more rings wherein such rings may be attached together in a pendent manner or may be fused, wherein the aromatic group has at least one heteroatom. Monocyclic heteroaromatic groups may contain 4 to 10 member atoms, typically 4 to 7 member atoms, and more typically 4 to 6 member atoms in the ring. Typical polycyclic heteroaromatic groups have two or three rings. Polycyclic aromatic groups having two rings typically have 8 to 12 member atoms, more typically 8 to 10 member atoms in the rings. Examples of heteroaromatic groups include, but are not limited thereto, pyrrole, imidazole, thiazole, oxazole, furan, thiophene, triazole, pyrazole, isoxazole, isothiazole, pyridine, pyrazine, pyridazine, pyrimidine, triazine, indole, benzofuran, benzothiophene, benzimidazole, benzthiazole, quinoline, isoquinoline, quinazoline, quinoxaline and the like.

[0111] The terms "carboxy” group or "carboxyl” group, whether used alone or with other terms, such as "carboxyalkyl” group, denotes -(C=O)-O-, which includes any suitable “ester” groups(-R’-(C=0)-0-R, where the R, R’ are the same or different and may be any suitable groups).

[0112] The term "carbonyl” group, whether used alone or with other terms, such as "aminocarbonyl” group, denotes -(C=O)-.

[0113] The term “amide” group describes a — C(=O) — NRR”, where R, R” may be the same or different and are any suitable groups.

[0114] The term "alkylaminoalkyl” group encompasses aminoalkyl groups having the nitrogen atom independently substituted with an alkyl group. In certain embodiments, the alkylaminoalkyl groups are "loweralkylaminoalkyl" groups having alkyl groups of one to six carbon atoms. In other embodiments, the lower alkylaminoalkyl groups have alkyl groups of one to three carbon atoms. Suitable alkylaminoalkyl groups may be mono or dialkyl substituted, such as N-methylaminomethyl, N, N-dimethyl-aminoethyl, N, N- diethylaminomethyl and the like.

[0115] The term "alkylamidoalkyl” group encompasses amidoalkyl groups having the nitrogen atom of the amide independently substituted with an alkyl group. In certain embodiments, the alkylamidoalkyl groups are "loweralkylamidoalkyl" groups having alkyl groups of one to six carbon atoms. In other embodiments, the lower alkylamidoalkyl groups have alkyl groups of one to three carbon atoms. Suitable alkylamidoalkyl groups may be mono or dialkyl substituted, such as N-methylamidomethyl, N, N-dimethyl-amidoethyl, N, N-diethylamidomethyl and the like.

[0116] The term "aralkyl” group encompasses aryl-substituted alkyl groups. In embodiments, the aralkyl groups are "lower aralkyl" groups having aryl groups attached to alkyl groups having one to six carbon atoms. In other embodiments, the lower aralkyl groups phenyl is attached to alkyl portions having one to three carbon atoms. Examples of such groups include benzyl, diphenylmethyl and phenylethyl. The aryl in said aralkyl may be additionally substituted with halo, alkyl, alkoxy, haloalkyl and haloalkoxy.

[0117] The term "alkylamino” group denotes amino groups which have been substituted with one alkyl group and with two alkyl groups, including terms "N-alkylamino" and "N,N- dialkylamino". In embodiments, alkylamino groups are "lower alkylamino" groups having one or two alkyl groups of one to six carbon atoms, attached to a nitrogen atom. In other embodiments, lower alkylamino groups have one to three carbon atoms. Suitable "alkylamino" groups may be mono or dialkylamino such as N-methylamino, N-ethylamino, N,N-dimethylamino, N,N-diethylamino and the like.

[0118] The term "suitable substituent", "substituent" or "substituted" used in conjunction with the groups disclosed herein refers to a chemically and pharmaceutically acceptable group, i.e., a moiety that does not negate the therapeutic activity of the inventive compounds. It is understood that substituents and substitution patterns on the compounds of the invention may be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques known in the art, as well as those methods set forth below. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon / member atom or on different carbons / member atoms, as long as a stable structure results. Illustrative examples of some suitable substituents include, cycloalkyl, heterocyclyl, hydroxyalkyl, benzyl, carbonyl, halo, haloalkyl, perfluoroalkyl, perfluoroalkoxy, alkyl, alkenyl, alkynyl, hydroxy, oxo, mercapto, alkylthio, alkoxy, aryl or heteroaryl, aryloxy or heteroaryloxy, aralkyl or heteroaralkyl, aralkoxy or heteroaralkoxy, HO-(C=O)--, amido, amino, alkyl- and dialkylamino, cyano, nitro, carbamoyl, alkylcarbonyl, alkoxycarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, arylcarbonyl, aryloxycarbonyl, alkylsulfonyl, and arylsulfonyl. Typical substituents include aromatic groups, substituted aromatic groups, hydrocarbon groups including alkyl groups such as methyl groups, substituted hydrocarbon groups such as benzyl, and heterogeneous groups including alkoxy groups such as methoxy groups.

[0119] The term "substituted" used in conjunction with the groups disclosed herein refers to a chemically acceptable group, i.e., a moiety that does not negate the activity of the surfactants. It is understood that substituents and substitution patterns on the surfactants may be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques known in the art. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon / member atom or on different carbons / member atoms, as long as a stable structure results. Illustrative examples of some suitable substituents include, for example, cycloalkyl, heterocyclyl, hydroxyalkyl, benzyl, carbonyl, halo, haloalkyl, perfluoroalkyl, perfluoroalkoxy, alkyl, alkenyl, alkynyl, hydroxy, oxo, mercapto, alkylthio, alkoxy, aryl or heteroaryl, aryloxy or heteroaryloxy, aralkyl or heteroaralkyl, aralkoxy or heteroaralkoxy, HO-(C=O)--, amido, amino, alkyl- and dialkylamino, cyano, nitro, carbamoyl, alkylcarbonyl, alkoxycarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, arylcarbonyl, aryloxycarbonyl, alkylsulfonyl, and arylsulfonyl.

[0120] In understanding the scope of the present application, the articles “a”, “an”, “the”, and “said” are intended to mean that there are one or more of the elements.

[0121] The term "comprising" and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The foregoing also applies to words having similar meanings such as the terms, "including", "having" and their derivatives. It will be understood that any embodiments described as “comprising” certain components may also “consist of’ or “consist essentially of,” wherein “consisting of’ has a closed-ended or restrictive meaning and “consisting essentially of’ means including the components specified but excluding other components except for materials present as impurities, unavoidable materials present as a result of processes used to provide the components, and components added for a purpose other than achieving the technical effect of the invention. For example, a composition defined using the phrase “consisting essentially of’ encompasses any known pharmaceutically acceptable additive, excipient, diluent, carrier, and the like. Typically, a composition consisting essentially of a set of components will comprise less than 5% by weight, typically less than 3% by weight, more typically less than 1 % by weight of non-specified components.

[0122] It will be understood that any component defined herein as being included may be explicitly excluded from the claimed invention by way of proviso or negative limitation, such as any specific compounds or method steps, whether implicitly or explicitly defined herein.

[0123] In addition, all ranges given herein include the end of the ranges and also any intermediate range points, whether explicitly stated or not.

[0124] Finally, terms of degree such as "substantially", "about" and "approximately" as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree may be construed as including a deviation of at least ±10% of the modified term if this deviation would not negate the meaning of the word it modifies.

[0125] The abbreviation, “e.g.” is derived from the Latin exempli gratia, and is used herein to indicate a non-limiting example. Thus, the abbreviation “e.g.” is synonymous with the term “for example.” The word “or” is intended to include “and” unless the context clearly indicates otherwise.

[0126] The phrase “at least one of’ is understood to be one or more. The phrase “at least one of...and...” is understood to mean at least one of the elements listed or a combination thereof, if not explicitly listed. For example, “at least one of A, B, and C” is understood to mean A alone or B alone or C alone or a combination of A and B or a combination of A and C or a combination of B and C or a combination of A, B, and C.

[0127] The phrase “a combination thereof’ in conjunction with a list of two or more features, for example, “A, B, C, or a combination thereof’ is understood to mean at least one of the elements listed or a combination thereof, if not explicitly listed. For example, A alone or B alone or C alone or a combination of A and B or a combination of A and C or a combination of B and C or a combination of A, B, and C.

[0128] It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting. Patent applications, patents, and publications are cited herein to assist in understanding the aspects described. All such references cited herein are incorporated herein by reference in their entirety and for all purposes to the same extent as if each individual publication or patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material. I. Conjugate Compounds

[0129] In general embodiments, there is provided a conjugate compound comprising: TTA-X1-MNP-X2-TpA.

[0130] TTA is a tumour targeting agent; MNP is a metal nanoparticle; TpA is a therapeutic agent; and X1and X2are the same or different. X1and X2are each independently selected from a non-covalent linker or a covalent linker.

[0131] Disclosed herein is a conjugate compound, a composition thereof, and a method for making the same. The conjugate compound and / or the composition thereof may improve therapeutic outcomes while inhibiting side effects and advancing the field of precision medicine.

[0132] With respect to the MNPs, TTAs, TpAs, and linkers X1and X2, embodiments are provided below. The embodiments of MNPs, TTAs, TpAs, and linkers X1and X2provided may be used in any suitable combination.

[0133] TTA

[0134] Tumour targeting agent (TTA) can refer to any suitable substance that is capable of specifically binding to or recognizing a target cell such as, and without being limited thereto, cancer cells (e.g. tumour cells). In this way, the TTA recognizes or binds to cancer cells, such as the surface of a tumour or in a tumour. With respect to the surface of a tumour, the TTA recognizes or binds to cancer cells, such as tumour derived peptides presented on the surface of tumour cells or in the tumor microenvironment. The TTA can be, for example, a glycan, a peptide, a protein, a polypeptide, a glycoprotein, a monosaccharide, a polysaccharide, an antibody or functional equivalent thereof, an antibody fragment, antibody mimic, nucleic acids, nucleosides, aptamers, cell receptor inhibitors, hormones, vitamin, synthetic molecules and the like. For example, aptamers and binding peptides have binding capabilities that mimic the action of antibodies. There are also ligands such as hormones and cytokines that can target cellular receptors on tumour cells; and there are substances such as folic acid and transferrin that are preferentially taken up by tumours. While the TTA can be any of the foregoing, typically, TTA is an antibody, or a functional equivalent thereof, that is capable of recognizing or binding to the tumour so that the conjugate compound disclosed herein may target the tumour.

[0135] In specific embodiments, the TTA is an antibody (or functional equivalent thereof). The antibody can be any suitable antibody, including but not limited to, a commercially available antibody available from a variety of sources including but not limited to: HyTest (Turku, Finland), InvivoGen (California, USA), BioRad (California, USA), Novus Biologicals (Colorado, USA), and Meridian Life Sciences (Tennessee, USA). In embodiments, the antibody is selected from, for example, monoclonal, polyclonal or multispecific antibodies, multivalent antibodies, minibodies, heteroconjugates, diabodies, triabodies, chimeric antibodies, synthetic antibodies, humanized antibodies, antibody fragments, and binding agents that employ the complementarity determining regions (CDRs) (or variants thereof that retain antigen binding activity) of the parent antibody. In some embodiments, the antibody or functional equivalent thereof is anti-CD20, anti-PSMA, anti-CD47, anti-EGFR or a combination thereof. In some embodiments, the antibody of the TTA may also be one of the chemotherapeutic drug molecule antibodies. For example, cetuximab or panitumumab. The TTA may also comprise a natural or synthetic flanking functional group, such as an amine group, a carboxylic acid, or ester group, azide for conjugation with the conjugate compound(s).

[0136] In other embodiments, at least a portion of the TTA may have a nucleic acid / nucleotide that is a CRG that complements with at least a portion of another CRG to form a covalent linker, as described below.

[0137] Exemplary tumour antigens that may be targeted by the TTA include 1 GH-IGK, 43- 9F, 5T4, 791Tgp72, acyclophilin C-associated protein, alpha-fetoprotein (AFP), a-actinin-4, A3, antigen specific for A33 antibody, ART-4, B7, Ba 733, BAGE, BCMA, BCR-ABL, beta- catenin, beta-HCG, BrE3-antigen, BCA225, BTAA, CA125, CA 15-3\CA 27.29\BCAA, CA195, CA242, CA-50, CAM43, CAMEL, CAP- 1 , carbonic anhydrase IX, c-Met, CA19-9, CA72-4, CAM 17.1 , CASP-8 / m, CCCL19, CCCL21 , CD1 , CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, CD20, CD21 , CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD44, CD45, CD46, CD47, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD68, CD70, CD70L, CD74, CD79a, CD79b, CD80, CD83, CD95, CD126, CD132, CD133, CD138, CD147, CD154, CDC27, CDK4, CDK4m, CDKN2A, CO-029, CTLA4, CXCR4, CXCR7, CXCL12, HIF-1a, colon-specific antigen-p (CSAp), CEA (CEACAM5), CEACAM6, c-Met, DAM, E2A-PRL, EGFR, EGFRvlll, EGP-1 (TROP-2), EGP-2, ELF2-M, Ep-CAM, fibroblast growth factor (FGF), FGF-5, Flt-1 , Flt-3, folate receptor, G250 antigen, Ga733VEpCAM, GAGE, gp1OO, GRO- , H4-RET, HLA-DR, HM1.24, human chorionic gonadotropin (HCG) and its subunits, HER2 / neu, HMGB-1 , hypoxia inducible factor (HIF-1), HSP70-2M, HST-2, HTgp-175, la, IGF-1 R, IFN- y, IFN-a, IFN-P, IFN-A, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-2, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-23, IL-25, insulin-like growth factor-1 (IGF-1), KC4-antigen, KSA, KS- 1-antigen, KS1-4, LAGE-1a, Le-Y, LDR / FUT, M344, MA-50, macrophage migration inhibitory factor (MIF), MAGE, MAGE-1 , MAGE-3, MAGE-4, MAGE-5, MAGE-6, MART-1 , MART-2, TRAG-3, mCRP, MCP-1 , MIP-1A, MIP-1 B, MIF, MG7-Ag, M0V18, MUC1 , MUC2, MUC3, MUC4, MUC5ac, MUC13, MUC16, MUM-1 / 2, MUM-3, MYL-RAR, NB / 70K, Nm23H1 , NuMA, NCA66, NCA95, NCA90, NY-ESO-1 , p15, p16, p185erbB2, p180erbB3, PAM4 antigen, pancreatic cancer mucin, PD1 receptor (PD-1), PD-1 receptor ligand 1 (PD- L1), PD-1 receptor ligand 2 (PD-L2), PI5, placental growth factor, p53, PLAGL2, Pmel17 prostatic acid phosphatase, PSA, PRAME, PSMA, P1GF, ILGF, ILGF-1R, IL-6, IL-25, RCAS1 , RS5, RAGE, RANTES, Ras, T101 , SAGE, S100, survivin, survivin-2B, SDDCAGi6, TA-90Mac2 binding protein, TAAL6, TAC, TAG-72, TLP, tenascin, TRAIL receptors, TRP-1 , TRP-2, TSP-180, TNF-a, Tn antigen, Thomson-Friedenreich antigens, tumor necrosis antigens, tyrosinase, VEGFR, ED-B fibronectin, WT-1 , 17-1A-antigen, complement factors C3, C3a, C3b, C5a, C5, an angiogenesis marker, bcl-2, bcl-6, K-ras, or any combination thereof.

[0138] MNP

[0139] The metal nanoparticle (MNP) can have an average particle size that is less than about 1 m in size, and typically from about 1 nm to about 900 nm in size and various ranges therebetween. In most embodiments, the average particle size is about 1 nm to about 100 nm. The particle can have one or more dimensions of the order of 100 nm or less. A nanoparticle can be made of a variety of materials, including but not limited to, transition metals or precious metals and are typically selected from gold, nickel, palladium, platinum, copper, silver, zinc, cadmium, or a combination thereof. With respect to combinations of metals, the metal nanoparticle can be a metal alloy (combines more than one metal or mixes a metal with other non-metallic elements). A metal nanoparticle can take a variety of shapes, including but not limited to, tube, rod, shell, cage, sphere, fiber, wire, stars, plate, sea archon, or a combination thereof. As examples, nanofibers are fibers with diameters less than 100 nanometers; nanowires are about 75 nm in diameter, and range from 1 pm to 10 microns in length; nanotubes are cylindrical nanoscale structures with length-to-diameter aspect ratios of up to 132,000:1.

[0140] In typical embodiments, the MNP is a metal nanorod (MNR). Dimensions of an MNR usually ranges from about 1 to about 100 nm. Typically the MNR has a diameter or cross-section of between about 5 nm and about 50 nm, such as from about 5 nm, about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, or about 45 nm, to about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, about 45 nm, or about 50 nm. For example, the diameter may be from about 5 nm to about 30 nm or from about 15 nm to about 30 nm. The MNR may typically have an axial length of between about 20 nm and about 500 nm, such as from about 20 nm, about 30 nm, about 40 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 125 nm, about 150 nm, about 175 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, or about 450 nm, to about 30 nm, about 40 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 125 nm, about 150 nm, about 175 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, or about 500 nm. For example, the axial length may be from about 30 nm to about 500 nm, from about 50 nm to about 300 nm, or from about 80 nm to about 100 nm.

[0141] Furthermore, the MNR may typically have an aspect ratio (i.e., the ratio of the length of the major axis of the nanorod to the minor axis of the nanorod) of from about 1.1 to about 100, such as from about 1.1 , about 1.2, about 1.3, about 1 .4, about 1.5, about 1 .6, about 1.7, about 1 .8, about 1 .9, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, or about 90, to about 1.2, about 1 .3, about 1 .4, about 1.5, about 1 .6, about 1 .7, about 1 .8, about 1.9, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 100. For example, the aspect ratio may be from about 1.1 to about 10. Substantially uniform length, diameter, and / or aspect ratio is used to refer to a population of metal nanorods wherein a majority of the metal nanorods have the same length, diameter, and / or aspect ratio within an acceptable variance for a subsequent analysis of the population. The population can be a single population in a sample or a subpopulation within a sample. In particular embodiments, the acceptable variance for the length, diameter, and / or aspect ratio of any given metal nanorod in the population or subpopulation can be at most 10%, 8%, 5%, 2%, 1 % or 0.1% different from the average length, diameter, and / or aspect ratio for metal nanorods in the population. In embodiments, the population can be composed of at least 90%, 95%, 99% or 99.9% metal nanorods having a particular length, diameter, and / or aspect ratio.

[0142] In embodiments, at least a portion of the surface of the nanorod is substantially smooth. In embodiments, at least a portion of the surface of the nanorod is substantially smooth and / or at least a portion of the surface may be etched (e.g. symmetrically etched and / or asymmetrically etched). In certain embodiments, the nanorod may be symmetrically etched to provide a multi-harmonic shape (e.g. appears as wave in 2-D).

[0143] MNRs may be synthesized from metals or semiconducting materials or their combinations. A MNR can have two ends and a linear body between the two ends. The two ends are also called the transverse or shorter ends. Accordingly, the longitudinal surface of the linear body is also called the longitudinal or longer end. The methods, compositions, and MNRs disclosed herein have been exemplified with respect to gold as the metal, however, it will be understood that the methods are equally applicable to nanorods of other metals, particularly those listed above. Known methods may be used to make the metal nanorods such as, and without being limited thereto, those described in WO2019 / 084661 , which is incorporated by reference in its entirety. MNPs that are compact in size, biocompatible, have tunable optical properties, efficient at conversion of light to heat, and / or capable of functionalization are typically used, and in particular, gold nanorods (GNRs).

[0144] In general, the MNP (e.g. MNR) can be bare, or can be capped with any suitable capping agents such as, and without being limited thereto, carboxylic acid, conventional citrate, and / or a positively charged ligand. These capping agents can readily be replaced with covalent and charge chemistries. The term “capping agent” refers to a chemical entity that is adsorbed on the surface of MNP and can provide stability against substantial aggregation of nanoparticles.

[0145] TpA

[0146] The therapeutic agent (TpA) can refer to any substance having a therapeutic property that can produce a desired, usually beneficial, effect. In embodiments, TpAs can be any agent for treating a symptom or disease in an individual in need of such treatment. TpAs may treat, ameliorate, and / or prevent disease. TpAs, as disclosed herein, may be biologies or small molecules (e.g. synthetic molecules), or a combination thereof. TpAs can be any suitable molecule / compound that can provide a therapeutic benefit to the subject. The therapeutic benefit can be the treatment of cancer the prevention of further cancer development or metastasis.

[0147] The TpA may be selected from proteins, peptides, nucleic acids, amino acids, nucleosides, antibodies, antibody drug conjugates (ADC), antibody fragments, antibody ligands, peptide nucleic acids, small organic molecules, lipids, hormones, drugs, enzymes, lectin, cell adhesion molecule, antibody epitope, enzyme substrates, enzyme inhibitors, vitamins, glycan, inhibitor, apoenzyme, cofactor, coenzymes, organic molecules, carbohydrates, such as polysaccharides and monosaccharides, or a combination thereof.

[0148] In embodiments, the TpA is an immunomodulator. The immunomodulator can be any molecule that can modulate the immune response. It can be any suitable molecule, such as, proteins, peptides, nucleic acids, amino acids, nucleosides, antibodies, antibody fragments, antibody ligands, peptide nucleic acids, small organic molecules, lipids, hormones, glycan, drugs, enzymes, lectin, cell adhesion molecule, antibody epitope, enzyme substrates, enzyme inhibitors, coenzymes, inorganic molecules, carbohydrates, such as polysaccharides and monosaccharides, or a combination thereof. In some embodiments, the molecule is a protein. In typical embodiments, the immunomodulator is a cytokine, cytokine agonists, a chemokine, chemokine agonists, or a toll-like receptor (TLR) agonist. In certain embodiments, the immunomodulator is a cytokine agonist, cytokine, such as IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12 IL-15, IL-17, IL-23, IL-10, TNF-a, IFN-a, IFN- 0, IFN-y, cytokine agonist thereof, or a combination thereof. In other embodiments, the immunomodulator is a chemokine agonist, chemokine, such as CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CXCL11 , CXCL12, CXCL14, CCL2 and CCL5, chemokine agonist thereof, or a combination thereof. In other embodiments, the immunomodulator comprises a checkpoint inhibitor, such as a PD1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, or any combination thereof. In some cases, the PD1 inhibitors comprise monoclonal antibodies that specifically target and block the PD1 protein. Examples of PD1 inhibitors include nivolumab (Opdivo™) (approved for various cancers, including melanoma, lung cancer, kidney cancer, and lymphoma); pembrolizumab (Keytruda™) (approved for various cancers, including non-small cell lung cancer, melanoma, and bladder cancer); and cemiplimab (Libtayo™) (approved for cutaneous squamous cell carcinoma). Ipilimumab is an exemplary CTLA-4 inhibitor approved for use in advanced melanoma and advanced renal cell cancer. Atezolizumab (for lung cancer, liver cancers, breast cancers, and urothelial bladder cancer), Avelumab (for merkel cell carcinoma (MCC), kidney cancer, or urothelial bladder cancer), and Durvalumab (for NSCLC and bile duct cancer) are exemplary PD-L1 inhibitors. Relatlimab is an exemplary LAG-3 checkpoint inhibitor, which is typically available mixed together with nivolumab for treatment of advanced melanoma. These exemplary checkpoint inhibitors can be used off label for treating other cancers, as will be understood.

[0149] In further embodiments, the immunomodulator is a TLR agonist. The TLR agonists, include, for example, agonists of TLR1 , TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10 and TLR11 , and in typical embodiments, the TLR agonists comprise TLR-4 agonists, TLR-7 agonists, TLR-8 agonists, TLR-9 agonists, or a combination thereof. Examples of TLR agonists, include but not are limited to, lefitolimod, tilsotolimod, rintatolimod, DSP-0509, AL-034, G-100, cobitolimod, AST-008, motolimod, GSK-1795091 , GSK-2245035, VTX-1463, GS-9688, LHC-165, BDB-001 , RG-7854, telratolimod. In specific embodiments, the TLR2 agonists, include but are not limited to, lipoteichoic acid, atypical LPS, MALP-2 and MALP-404, OspA, porin, LcrV, lipomannan, GPI anchor, lysophosphatidylserine, lipophosphoglycan (LPG), glycophosphatidylinositol (GPI), zymosan, hsp60, gH / gL glycoprotein, hemagglutinin; the TLR-3 agonist includes, but is not limited to, double-stranded RNA, (e.g., poly(l:C)), rintatolimod, poly-ICLC, RIBOXXON™, Apoxxim, RIBOXXIM™, IPH-33, MCT-465, MCT-475, and ND-1.1 ; the TLR-4 agonists include, but is not limited to, lipopolysaccharides (LPS), lipoteichoic acid, (3-defensin 2, fibronectin EDA, HMGB1 , snapin, tenascin C, monophosphoryl lipid A (MPLA), G100, GSK1795091 , and PEPA-10; the TLR5 agonists, include, but are not limited to, flagellin; the TLR7 / TLR8 agonists include, but are not limited to single-stranded RNA, CpG-A, Poly G10, Poly G3, Resiquimod (R848), 3M-052), NKTR-262, IMO-4200, MEDI-9197 (telratolimod); TLR7 agonists, such as DS-0509, GS-9620, LHC-165, TMX-101 (imiquimod); TLR-8 agonists include, but are not limited to, MCT-465, motolimod, GS-9688, and VTX-146; TLR-9 agonists include, but are not limited to, unmethylated CpG DNA, AST-008, IMO-2055, IMO-2125, lefitolimod, litenimod, MGN-1601 , and PUL-042. Nonlimiting examples of TLR agonists can be found at W02008115319, US20130202707, US20120219615, US20100029585, W02009030996, W02009088401 , and WO2011044246, each of which is incorporated by reference in its entirety.

[0150] In embodiments, the TpA is a drug, such as a chemotherapeutic drug or agent. The chemotherapeutic drug or agent may encompass any non-proteinaceous (e.g., non- peptidic) chemical compound useful in the treatment of cancer. Examples of such chemotherapeutic agents include, but not limited to, alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN™); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodepa, carboquone, meturedepa, and uredepa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimemylolomelamine; acetogenins, e.g., bullatacin and bullatacinone; a camptothecin, including synthetic analog topotecan; bryostatin, callystatin; CC-1065, including its adozelesin, carzelesin, and bizelesin synthetic analogs; cryptophycins, particularly cryptophycin 1 and cryptophycin 8; dolastatin; duocarmycin, including the synthetic analogs KW-2189 and CBI-TMI; eleutherobin; 5-azacytidine; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cyclophosphamide, glufosfamide, evofosfamide, bendamustine, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, and uracil mustard; nitrosoureas such as carmustine, chlorozotocin, foremustine, lomustine, nimustine, and ranimustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gammall and calicheamicin phil 1), dynemicin including dynemicin A, bisphosphonates such as clodronate, an esperamicin, neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromomophores, aclacinomycins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, carrninomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholinodoxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, and zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as demopterin, methotrexate, pteropterin, and trimetrexate; purine analogs such as cladribine, pentostatin, fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, and testolactone; antiadrenals such as aminoglutethimide, mitotane, and trilostane; folic acid replinishers such as frolinic acid; radiotherapeutic agents such as Radium-223, 177-Lu-PSMA-617; trichothecenes, especially T-2 toxin, verracurin A, roridin A, and anguidine; taxoids such as paclitaxel (TAXOL™), albumin-bound or nab-paclitaxel (ABRAXANE™), docetaxel (TAXOTERE™), cabazitaxel, BIND-014, tesetaxel; platinum analogs such as cisplatin and carboplatin, NC-6004 nanoplatin; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; hestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elformthine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; leucovorin; lonidamine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidamol; nitracrine; phenamet; pirarubicin; losoxantrone; fluoropyrimidine; folinic acid; podophyllinic acid; 2-ethylhydrazide; procarbazine; polysaccharide-K (PSK); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; trabectedin, triaziquone; 2,2',2"-trichlorotriemylamine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiopeta; chlorambucil; gemcitabine (GEMZAR™); 6- thioguanine; mercaptopurine; methotrexate; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitroxantrone; vancristine; vinorelbine (NAVELBINE™); novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeoloda; ibandronate; CPT-11 ; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DFMO); retinoids such as retinoic acid; capecitabine; NUC-1031 ; FOLFOX (folinic acid, 5-fluorouracil, oxaliplatin); FOLFIRI (folinic acid, 5-fluorouracil, irinotecan); FOLFOXIRI (folinic acid, 5-fluorouracil, oxaliplatin, irinotecan), FOLFIRINOX (folinic acid, 5-fluorouracil, irinotecan, oxaliplatin), and pharmaceutically acceptable salts, acids, or derivatives of any of the above.

[0151] In embodiments, the chemotherapeutic agent or drug can be, but are not limited to, anti-hormonal agents such as anti-estrogens and selective estrogen receptor modulators (SERMs), inhibitors of the enzyme aromatase, anti-androgens, and pharmaceutically acceptable salts, acids or derivatives thereof that act to regulate or inhibit hormone action on cancer cells and / or tumours. Examples of anti-estrogens and SERMs include, for example, tamoxifen (including NOLVADEX™), raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (FARESTON™). Examples of inhibitors of the enzyme aromatase, which regulate estrogen production in the adrenal glands, include, but are not limited to, 4(5)-imidazoles, aminoglutethimide, megestrol acetate (MEGACE™), exemestane, formestane, fadrozole, vorozole (RIVISOR™), letrozole (FEMARA™), and anastrozole (ARIMIDEX™). Examples of anti-androgens include, but are not limited to, apalutamide, abiraterone, enzalutamide, flutamide, galeterone, nilutamide, bicalutamide, leuprolide, goserelin, ODM-201 , APC-100, ODM-204. As would be understood, any of the non-proteinaceous (e.g., non-peptidic) chemical compounds or anti-hormonal agents listed herein may be conjugated onto an antibody to create an antibody-drug conjugate (ADC) which can be used as the therapeutic agent of the conjugate compound disclosed herein.

[0152] In specific embodiments, the chemotherapeutic drug or agent is selected from, for example, Temozolomide, Actinomycin, Alitretinoin, All-trans retinoic acid, Azacitidine, Azathioprine, Bevacizumab, Bexatotene, Bleomycin, Bortezomib, Carboplatin, Capecitabine, Cetuximab, Cisplatin, Chlorambucil, Cyclophosphamide, Cytarabine, Daunorubicin, Docetaxel, Doxifluridine, Doxorubicin, Epirubicin, Epothilone, Erlotinib, Etoposide, Fluorouracil, Gefitinib, Gemcitabine, Hydroxyurea, Idarubicin, Imatinib, Ipilimumab, Irinotecan, Mechlorethamine, Melphalan, Mercaptopurine, Methotrexate, Mitoxantrone, Ocrelizumab, Ofatumumab, Oxaliplatin, Paclitaxel, Panitumab, Pemetrexed, Rituximab, Tafluposide, Teniposide, Tioguanine, Topotecan, Tretinoin, Valrubicin, Vemurafenib, Vinblastine, Vincristine, Vindesine, Vinorelbine, Vorinostat, Romidepsin, 5- fluorouracil (5-FU), 6-mercaptopurine (6-MP), Cladribine, Clofarabine, Floxuridine, Fludarabine, Pentostatin, Mitomycin, ixabepilone, Estramustine, prednisone, methylprednisolone, dexamethasone, or a combination thereof.

[0153] At least a portion of the TpA may have a nucleic acid / nucleotide that complements with a portion of the covalent linker to form the covalent linker, as described below.

[0154] In other embodiments, at least a portion of the TpA may have a nucleic acid / nucleotide that is a CRG that complements with at least a portion of another CRG to form a covalent linker, as described below.

[0155] Linkers X1and X2

[0156] X1and X2are the same or different and are each independently selected from a non-covalent linker or a covalent linker and may be cleavable. In other embodiments, at least one of X1and X2is a cleavable linker. In examples, either X1or X2is a cleavable linker such that either X1or X2is cleaved to release the TpA or the TTA, typically, release the TpA. a) In embodiments of the non-covalent linker, the non-covalent linker can refer to a non-covalent bond (e.g., electrostatic interactions (e.g., ionic bond, hydrogen bond, etc.) or van der Waals interactions (e.g., dipole-dipole, dipole-induced dipole, London dispersion, etc.) between two moieties. For example, the non-covalent linker is the result of two molecules (e.g. TTA and the MNP) that are not covalently linked to each other and interact with each other via a non-covalent bond. The non-covalent linker can be the result of the association between atoms or molecules of conjugate reactive groups (CRGs). The CRG can refer to a moiety or group capable of forming a non-covalent linker as a result of the association between atoms or molecules (e.g. ssDNA). The association can be direct or indirect. CRGs can be, for example, complementary ssDNA pair (e.g. ssDNA(1 ’) is CRG1 and ssDNA(1”) is CRG2), wherein the non-covalent linker, DNA, is formed from H- bonding of the complementary ssDNA(1 ’) and ssDNA (1”).

[0157] In embodiments, the non-covalent linker can be at least one base pair of nucleotides. In embodiments, the non-covalent linker is at least two base pairs of nucleotides. For example, there may be at most 50 base pairs of nucleotides, at most 40 base pairs of nucleotides, at most 30 base pairs of nucleotides, at most 20 base pairs of nucleotides, or about 2 to about 20 base pairs of nucleotides. Therefore, the non-covalent linker can be a complementary polynucleotide or oligonucleotide. In other embodiments, it can be DNA and / or RNA with a suitable sequence and length. The non-covalent linker can be the result of two complementary ssDNA, mRNA, XNA, etc.

[0158] In embodiments, the non-covalent linker can result from combining an antibody and MNPs (e.g. GNRs). b) In embodiments of the covalent linker, the covalent linker can refer to a divalent moiety, which connects at least two molecules (e.g. TTA and the MNP) via a linking covalent molecule or a direct bond. In embodiments, a functional group (or zero length linking group) can result from the direct reaction of CRGs or the addition of a further linker (e.g. crosslinker, chain length of at least one) of the CRGs. Embodiments of methods for the like can be found, for example, in Hermanson, G.T., “Bioconjugation Techniques”, 3rd edition, Chapter 3, pages 229-258, and is incorporated by reference. Some functional groups that may be used in conjugation herein are as follows:

[0159] 1. Amine reactive with Isothiocyanate, Isocyante, Acyl azide, NHS easter, Sulfonyl Chloride, Tosyl easter, Tresyl easter, aldehyde, amine, epoxide, carbonate, aryl halide, Haloacetyl or alkyl halide, Imido easter, Carboxylate, Alkyl phosphate, Anhydride, Fluorophenyl easter, HOBt easter, Hydroxymethyl phosphine, O- methyl urea, DSC, NHS carbamate, Glutaraldehyde, active double bond, contrained triple bond, Cyclic hemiacetal, NHS carbomate, Imidazole carbamate, Acyl Imidazole, Methylpyridinium ether, Azolactone, Cyanate ester, Cyclic, imidocarbonate, Chlorotriazine, and Dehydroazepine 6-sulfo-cytosine derivative.

[0160] 2. Thiol reactive with Haloacetyl or alkyl halide, Malemide, Aziridine, Aryl halide, Pyridyl disulfide, 2,2-dipyridyl disulphide, 4,4-dipyridyl disulphide, TNB thiol, Ellman’s reagent, Peroxide, Vinylsulfone, Metal surface, phenylthioesters (on C- terminal peptides), Cisplatin, and Activated double bond.

[0161] 3. Carboxylate reactive with Diazoalkanes, and diazoacetyl compounds.

[0162] 4. Hydroxyl group reactive with Epoxide, Haloacetyl, alkyl halide, and Isocyanate.

[0163] 5. Aldehyde group reactive with Amine, Hydrazide, hydrazine, Aminooxy compound

[0164] 6. Active hydrogen compounds reactive with Amine, Diazonium, l2, Halogenated Phenyl azide, Phenyl azide, Benzophenone, Anthraquinone, Diazo derivatives, and Diazirine derivative.

[0165] 7. Thymine base reactive with Psorale derivative.

[0166] 8. Diene reactive with Aldehyde, aminooxy, Azide, and phenyl boronic acid.

[0167] 9. Alkyl phosphate (e.g. 5’-phosphate of oligonucleotide reactive with Carbodiiimide and imidazole.

[0168] 10. Diol cleavage to form two aldehydes.

[0169] 11 . Disulfide cleavage to form two sulfhydryls.

[0170] 12. Ester Bond cleavage to form a carboxylic and a hydroxyl groups.

[0171] 13. Diazo cleavage to form two aryl amines.

[0172] 14. Sulfone bond cleavage to form a sulfonate and a hydroxyl group.

[0173] 15. Acyl Hydrazone bond cleavage to form a hydrazone and a hydrazide.

[0174] 16. O-Nitrophenyl group cleavage to form a methyl ketone and an amine.

[0175] 17. Aryl amine conversion to Diazonium group.

[0176] Some embodiments of linkages formed from reaction of CRGs include:

[0177] 1 . Reaction between Aldehyde or Ketone with Carboxylic acid group: One CRG comprises a carbonyl group and another comprises a carboxylic acid group can come from either CRG. In an embodiment, a carboxylic acid on the MNPs (e.g. GNRs) surface can be established by using a modified thiol-PEG that has a carboxylic acid terminating group. In another embodiment, a small molecule, such as thiolated lipoic acid, can also be used to provide carboxylate groups on MNPs. Carboxylated MNPs can react with carbohydrazide in the presence of EDC (1-ethyl-3-(3-(dimethylaminopropyl)-carbodiimide) to produce hydrazide MNPs. These MNPs can then react with aldehyde- or ketone- containing molecules to produce a hydrazone bond (e.g. Schiff base that can be reduced with sodium cyanoborohydride) MNP-CO-NH-NH-TTA (or TpA). Another embodiment of a method that can be uses is similar to a protocol described in Hermanson, G.T., “Bioconjugation Techniques”, 3rdedition, pages 571-573 can be used for this purpose. Similarly, if a carboxylic acid group is from the TTA or TPA then MNPs modified with aldehyde or ketone can be used. Reaction between Amine and Carboxylic Acid group: This conjugation can be achieved using carbodiimide coupling. An embodiment of a method is described in Hermanson, G.T., “Bioconjugation Techniques”, 3rdedition, pages 262-267 or 944. This protocol can be used to form an amide bond between MNPs and TTA or TpA. MNPs (e.g. GNRs) can be functionalized to complement availability of an amine or carboxylic acid group of TTA or T pA. Reaction between an Amine and an Azide group: In an embodiment, MNPs (e.g. GNRs) are suspended in a solution of azo-PEG-thiol to modify GNRs with thiol-PEG-azide linker. TTA(or TpA)-NH2is reacted with 4-pentynoic acid in the presence of EDO to create alkynyl-labeled TTA or TpA. Alkynyl-labeled TTA or TpA and modified MNPs are combined (e.g. in the presence of Cu+) to form a conjugate. In another embodiment, a method similar to a protocol described in Hermanson, G.T., “Bioconjugation Techniques”, 3rdedition, pages 774-775 can be used. Reaction with NSH or sulfo-NHS: In an embodiment, MNPs (e.g. GNRs) have a carboxylic acid group, which is converted to an O-acylisourea intermediate and reacts with NSH or sulfo-NHS to establish an amide linkage. In another embodiment, a method similar to a protocol described in Hermanson, G.T., “Bioconjugation Techniques”, 3rdedition, pages 265-267 can be used. Primary amine modification with SATA (N-succinimidyl S-acetylthioacetate): In an embodiment, hydrolysis of an amine group from TTA or TpA and a sulfhydryl kit from ThermoFisher Scientific with reagent SATA, SATA-PEG4 (PEGylated N-succinimidyl S-acetylthioacetate), or SATP (N-succinimidyl-S- acetylthiopropionate) can be used to form sulfhydryl. The latter can react with MNPs (e.g. GNRs) to make gold-thiol linkage or with maleimide activated MNPs to make a linker through a thioether bond. In another embodiment, a method similar to a protocol described in Hermanson, G.T., “Bioconjugation Techniques”, 3rdedition, page 886 can be used. (See Figure 40) Examples of chemical groups that can react with primary amines:

[0178] 7. Glycan targeting: TpA or TTA that have a glycan moiety can be conjugated to an amine functionalized MNPs (e.g. GNRs) with periodate activation. In an embodiment, a method similar to a protocol described in Hermanson, G.T., “Bioconjugation Techniques”, 3rdedition 625-628 can be used.

[0179] Therefore, the covalent linker can be any linker molecule with at least one functional group (e.g. amide, carboxyl, disulfide, alkene, alkyne, etc.) that links the TTA to the MNP (e.g. GNR) and / or the TpA to the MNP. The covalent linker can also be the result of the association between atoms or molecules of CRGs, which can include a zero length linker or a bifunctional cross-linker. With respect to CRGs for covalent linkers, the CRG refers to a moiety or group capable of forming a covalent linker as a result of the association between atoms or molecules of each CRG, which can include a linking functional group. The association can be direct or indirect.

[0180] For examples of forming a covalent linker, a conjugate between a first CRG (e.g., - COOH, carboxylate, -N-hydroxysuccinimide, carbodiimide, or -maleimide) and a second CRG (e.g., sulfhydryl, sulfur-containing amino acid, amine, amine sidechain containing amino acid, carbodiimide, COOH, or carboxylate) can be direct, e.g., by covalent bond, or indirect, e.g., by non-covalent bond (e.g., electrostatic interactions (e.g., ionic bond, hydrogen bond, halogen bond) van der Waals interactions (e.g., dipole-dipole, dipole- induced dipole, London dispersion), ring stacking (pi effects), hydrophobic interactions and the like), for example, formation of DNA. In embodiments, the covalent linker is formed using conjugate chemistry (i.e., the association of two CRGs) including, but are not limited to nucleophilic substitutions (e.g., reactions of amines and alcohols with acyl halides, active esters), electrophilic substitutions (e.g., enamine reactions) and additions to carbon-carbon and carbon-heteroatom multiple bonds (e.g., Michael reaction, Diels-Alder addition). These and other useful reactions are discussed in, for example, March, ADVANCED ORGANIC CHEMISTRY, 3rd Ed., John Wiley & Sons, New York, 1985; Hermanson, CONJUGATE TECHNIQUES, Academic Press, San Diego, 1996; and Feeney et al., MODIFICATION OF PROTEINS; Advances in Chemistry Series, Vol. 198, American Chemical Society, Washington, D.C., 1982. In embodiments, the first CRG (e.g., carboxylate moiety) is covalently attached to the second CRG (e.g., an amine). In embodiments, the first CRG (e.g., maleimide moiety) is covalently attached to the second CRG (e.g., a sulfhydryl). In embodiments, the first CRG (e.g., haloacetyl moiety) is covalently attached to the second CRG (e.g., a sulfhydryl). In embodiments, the first CRG (e.g., pyridyl moiety) is covalently attached to the second CRG (e.g., a sulfhydryl). In embodiments, the first CRG (e.g., — N- hydroxysuccinimide moiety) is covalently attached to the second CRG (e.g., an amine). In embodiments, the first CRG (e.g., maleimide moiety) is covalently attached to the second CRG (e.g., a sulfhydryl). In embodiments, the first CRG (e.g., -sulfo-N-hydroxysuccinimide moiety) is covalently attached to the second CRG (e.g., an amine).

[0181] Useful CRGs used for conjugate chemistries herein include, for example: (a) carboxyl groups and various derivatives thereof including, but not limited to, N- hydroxysuccinimide esters (e.g. sulfo-N-hydroxysuccinimide), N-hydroxybenztriazole esters, acid halides, acyl imidazoles, thioesters, p-nitrophenyl esters, alkyl, alkenyl, alkynyl and aromatic esters; (b) hydroxyl groups which can be converted to esters, ethers, aldehydes, etc.; (c) haloalkyl groups wherein the halide can be later displaced with a nucleophilic group such as, for example, an amine, a carboxylate anion, thiol anion, carbanion, or an alkoxide ion, thereby resulting in the covalent attachment of a new group at the site of the halogen atom; (d) dienophile groups which are capable of participating in Diels-Alder reactions such as, for example, maleimido or maleimide groups; (e) aldehyde or ketone groups such that subsequent derivatization is possible via formation of carbonyl derivatives such as, for example, imines, hydrazones, semicarbazones or oximes, or via such mechanisms as Grignard addition or alkyllithium addition; (f) sulfonyl halide groups for subsequent reaction with amines, for example, to form sulfonamides; (g) thiol groups, which can be converted to disulfides, reacted with acyl halides, or bonded to metals such as gold, or react with maleimides; (h) amine or sulfhydryl groups (e.g., present in cysteine), which can be, for example, acylated, alkylated or oxidized; (i) alkenes, which can undergo, for example, cycloadditions, acylation, Michael addition, etc.; (j) epoxides, which can react with, for example, amines and hydroxyl compounds; (k) phosphoramidites and other standard functional groups useful in nucleic acid synthesis; (I) metal silicon oxide bonding; (m) metal bonding to reactive phosphorus groups (e.g., phosphines) to form, for example, phosphate diester bonds; (n) azides coupled to alkynes using copper catalyzed cycloaddition click chemistry; (o) biotin conjugate can react with avidin or strepavidin to form a avidin-biotin complex or streptavidin-biotin complex.

[0182] The covalent linker and the non-covalent linkers can be cleavable linkers. The cleavable linker may be cleavable in response to stimulus (e.g., photo-irradiation such as irradiation at certain wavelength(s) to control cleavage, for example and without being limited thereto, UV, visible, or NIR), enzymes, nucleophilic / basic reagents, reducing agents, electrophilic / acidic reagents, organometallic and metal reagents, or oxidizing reagents). Therefore, the cleavable linker can be, for example, one or more of a chemical cleavable linker, enzyme cleavable linker, heat cleavable linker, pH cleavable linker, and a photochemical cleavable linker. A chemical cleavable linker refers to a linker which is capable of cleaving in response to the presence of a chemical (e.g., acid, base, oxidizing agent, reducing agent, etc.). A chemical cleavable linker can be non-enzymatically cleavable.

[0183] Some embodiments of cleavable linkers that may be used are: i) Chemical cleavable linkers (e.g. hydrazones, disulfides, etc.): Hydrazone is a typical acid-sensitive (pH sensitive) linker. Hydrazone linked conjugates are generally stable in blood circulation but hydrolyzed to release the cytotoxic payloads in lysosome (pH 4.8) and endosome (pH5.5-6.2) upon internalization into the targeted cancer cells. Disulfide based linker can be sensitive to reductive glutathione (GSH). The concentration of GSH in blood is considerably lower than intracellular concentration in cancer cells. Therefore, this type of linker may be stable in blood system but specifically release the active payloads in the cancer cells with an elevated GSH. ii) Enzyme cleavable linkers (e.g. glucuronides, peptides, etc.): In terms of enzyme sensitive linkers, peptide based linkers may be sensitive to the lysosomal protease. The lysosomal proteases, such as cathepsin B, are generally overexpressed in cancer cells, enabling the accurate drug release in the vicinity of the tumour. Beta-glucuronide linker is another enzyme-sensitive linker commonly used in ADCs. It can be cleaved for payloads release in cells by beta-glucuronidase, the levels of which are often found higher in tumour regions. DNA is another example of an enzyme cleavable linker (e.g. cleavable with helicase). iii) Light / heat cleavable linker (e.g. heptamethine cyanine fluorophore triggerA = about 650 to about 900 nm), O-Nitrobenzyl trigger (NIR light, PC4AP trigger A = about 365 nm); nucleic acid melts such as dsDNA (heat) and dsRNA (heat).

[0184] Some examples of cleavable linkers are provided in Sheyi, R. et al., Linkers: An Assurance for Controlled Delivery of Antibody-Drug Conjugate, Pharmaceutics. 2022 Feb; 14(2): 396. (nih.gov): https: / / www.ncbi. nlm.nih.gov / pmc / articles / PMC8874516 / #:~:text=ADCs%20consist%20of% 20recombinant%20monoclonal,and%20pharmacokinetics%20of%20these%20drugs, incorporated by reference in its entirety: examples in Su, Zheng et al., Antibody-drug conjugates: Recent advances in linker chemistry - PMC; Acta Pharm Sin B. 2021 Dec; 11 (12): 3889-3907 (nih.gov) (https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC8727783 / ), incorporated by reference in its entirety:

[0185] Table 1

[0186] Chernies] triggers described in this review cBu-Cit trigger MMAE, PBD

[0187] Glycosidase jG-Glucuronide trigger Linkers cleavage by ?- MMAE cleavable triggers glucuronidase in lysosomes jG-Galactoside trigger Linkers cleavage by jG-galactosidase MMAE 36 in lysosomes

[0188] Phosphatase Pyrophosphate trigger Linkers cleavage by phosphatase Budesonide 18 cleavable triggers and pyrophosphates in lysosomes

[0189] Sulfatase cleavable Arylsulfate trigger Linkers cleavage by sulfatase in MMAE 37 trigger lysosomes

[0190] Photo-responsive Heptamethine cyanine Linkers cleavage by irradiation CA-4 46 cleavable triggers fluorophore trigger with NIR light [ = 650-900 nm)

[0191] O-Nitrobenzyl trigger Linkers cleavage by irradiation MMAE 47 with UV light [A - 365 n )

[0192] PC4AP trigger Linkers cleavage both by DOX 48 irradiation with near-infrared [NIR) light [A = 365 nm) and intramolecular addition reaction with nearby amine

[0193] Bioorthogonal dsProc trigger Linkers cleavage by the DOX 55 cleavable trigger bioorthogonal cleavage pair: Cu[I)- BTTAA / dsProc

[0194] Non-cleavable MD linker No linker cleavage, ADCs TRMRA 14 linkers PEG linkers with metabolizes amino acid appendage, PBD Dimer 58 intermediates of alkyne, a linker and molecule cytotoxicity triazole and piperazine upon entry lysosome

[0195] Mal-PAB linker MMAE 59

[0196] , and examples in Dubowchik, G.M. et al., Cathepsin B-Labile Dipeptide Linkers for Lysosomal Release of Doxorubicin from Internalizing Immunoconjugates: Model Studies of Enzymatic Drug Release and Antigen-Specific In Vitro Anticancer Activity, Bioconjugate Chemistry, 2002, 12, 4, 855-869 (acs.org) (https: / / pubs.acs.org / doi / 10.1021 / bc025536i'), incorporated by reference in its entirety.

[0197] Other cleavable linkers includes, but is not limited to, valine-citruline, valine-alanine and any combination of two to eight amino acids. A self-immolative unit (e.g., a PAB spacer) can be included to assist with clean cleavage, and optionally hydrophilic groups (e.g., PEG) can be added to increase hydrophilicity of the construct. In some aspects, more suitably, the linker group comprises a self-immolative unit. A range of self immolative units are known in the art

[0030] and have been described in, for example, U.S. Pat. No.

[0198] 7,754,681 , European Patent Publication No. 0624377. The maleimide methodology is routinely used as a method to attach antibodies to drug compounds by providing a linker attached to the drug with a terminal maleimide group. In addition, methodologies using diarylcyclooctyne moieties (such as, but not limited to, DBCO, dibenzylcyclooctyne) are known in the art. Diarylcyclooctynes react with stable azides to provide attachment via the formation of stable triazoles. Diarylcyclooctynes are thermostable with very narrow and specific reactivity toward azides, resulting in almost quantitative yields of stable triazoles. Furthermore, the reaction does not require a cytotoxic Cu(l) catalyst (that is toxic to most organisms) and thus, prevents its use in many biological systems. Still further, alkoxyamine methodologies are also alternatives in the art. For site-specific conjugation of the drug to the antibody, the antibodies may comprise a “tag” (which may be proprietary) that will react with a diarylcyclooctyne (for example DBCO), an alkyoxyamine and / or maleimide group to attach the antibody to the drug. The tag in some instances may be a mutated amino acid.

[0199] The cleavable linker can have a “cleavable site”, for example, the functional group (e.g. amide, disulfide, diselenide) as described for the covalent linker or the “unzipping” of DNA or the like for a non-covalent linker. A “cleavable site” or “scissile linkage” in the context of a polynucleotide is a site which allows controlled cleavage of the polynucleotide strand (e.g., the linker, the primer, or the polynucleotide) by chemical, enzymatic, or photochemical means known in the art and disclosed herein. A scissile site may refer to the linkage of a nucleotide between two other nucleotides in a nucleotide strand (i.e., an internucleosidic linkage). In embodiments, the scissile linkage can be located at any position within the one or more nucleic acid molecules, including at or near a terminal end (e.g., the 3' end of an oligonucleotide) or in an interior portion of the one or more nucleic acid molecules. In embodiments, conditions suitable for separating a scissile linkage include a modulating the pH and / or the temperature. In embodiments, a scissile site can include at least one acid-labile linkage. For example, an acid-labile linkage may include a phosphoramidate linkage. In embodiments, a phosphoramidate linkage can be hydrolysable under acidic conditions, including mild acidic conditions such as trifluoroacetic acid and a suitable temperature (e.g., 30° C), or other known conditions, for example Matthias Mag, et al. Tetrahedron Letters, Volume 33, Issue 48, 1992, 7319-7322. In embodiments, the scissile site can include at least one photolabile internucleosidic linkage (e.g., o-nitrobenzyl linkages, as described in Walker et al, J. Am. Chem. Soc. 1988, 110, 21 , 7170-7177), such as o-nitrobenzyloxymethyl or p-nitrobenzyloxymethyl group(s). In embodiments, the scissile site includes at least one uracil nucleobase. In embodiments, a uracil nucleobase can be cleaved with a uracil DNA glycosylase (UDG) or Formamidopyrimidine DNA Glycosylase Fpg. In embodiments, the scissile linkage site includes a sequence-specific nicking site having a nucleotide sequence that is recognized and nicked by a nicking endonuclease enzyme or a uracil DNA glycosylase.

[0200] In embodiments, the cleavable linker may be any suitable cleavable linker that can cleave in response to stimulus. The stimulus may be, and without being limited thereto, photo-irradiation, such as irradiation at certain wavelength(s) to control cleavage (e.g. UV, visible, or NIR), enzymes, nucleophilic / basic reagents, reducing agents, electrophilic / acidic reagents, organometallic and metal reagents, oxidizing reagents, or a combination thereof. The cleavable linker may be a chemical cleavable linker, enzyme cleavable linker, heat cleavable linker, pH cleavable linker, a photochemical cleavable linker, or a combination thereof. In a specific embodiment, the cleavable linker is a photochemical cleavable linker. This linker is cleavable via irradiation in the UV, visible, and / or NIR region.

[0201] The cleavable linker may be cleaved via irradiation at a wavelength of about 650 nm to about 1100 nm; about 700 nm to about 1100 nm; about 750 nm to about 1100 nm; about 800 nm to about 1100 nm; about 850 nm to about 1100 nm; about 700 nm to about 1000 nm; about 750 nm to about 950 nm; about 800 nm to about 1000 nm; or about 850 nm to about 1000 nm. With respect to the linkers, X1and X2, of the conjugate compound, disclosed herein, these may be used in any suitable combination. As described, X1and X2can be cleavable or non-cleavable linkers (e.g. labile or non-labile linkers). The linkers may be cleavable via a stimulus such as, and without being limited thereto, photo-irradiation (light / heat), pH, enzyme, etc. In embodiments, photo-irradiation may be provided from any suitable source of light. Various wavelengths can be used to both cleave the linkers and heat the MNP of the conjugate MNP. The MNP (e.g. GNR) may also heat and induce the cleavage of the linker(s) and consequent TpA release.

[0202] Various embodiments include: TTA is conjugated to the GNR using a non-labile linker in opposition to a labile linker. One of the two labile linker can be responsive to light, pH or an enzyme. TTA is conjugated to GNRs using a labile linker that is not DNA. GNRs are first conjugated to two ssDNA (from two sets of complementary DNA pairs) and then conjugated to TpA and TTA. The nucleic acid nature of some TpA is utilized in creation of double strand nucleic acid as labile linker. Conjugate compound can cleaves into two components only (GNR-TTA and TpA) upon irradiation of NIR and not three component (TTA, GNRs and TpA).

[0203] The conjugate compound can be protected from exposure to various elements, such as whole blood. Polymers can be used to protect the conjugate compound, such as synthetic polymers and / or natural polymers. Synthetic polymers are those suitable for use in mammals. Examples of synthetic polymers include PEG (polyethylene glycol), PVP (polyvinylpyrrolidone), PSS (polystyrene sulfonate), PLGA Poly lactic-co-glycolic acid, PNIPAM (poly(N-isopropylacrylamide)), ploxamer, diblock or triblock polymers, pH or thermo-responsive polymers, or a combination thereof. Natural polymers are those suitable for use in mammals. Examples of natural polymers include glycoproteins, albumin, gelatine, collagen, or a combination thereof. In addition to polymer(s) or separately, phospholipids (both low and high density), micelles, vesicles, liposomes, solid lipid nanoparticle (SLN), nanostructured lipid carriers (NLC), or a combination thereof. For the protection of the conjugate compound with polymer(s), a selected polymer may have a thiol or disulphide functional group (e.g. at a terminal end) or has been modified to include this functional group.

[0204] Nano-aggregates such as micelles, vesicles, liposomes, solid lipid nanoparticle (SLN), nanostructured lipid carriers (NLC) and / or supramolecular vesicles are described for encapsulation and / or surface adsorption of the conjugate compound for intra-tumoral and intravenous injection or as a topical application.

[0205] Representative coatings that may be used with the conjugate compounds disclosed herein and can include molecules having, for example, hydrophobic segments such as PPO segments with molecular weights of at least about 1 .8 kDa, or at least about 2 kDa, or at least about 2.4 kDa, or at least about 2.8 kDa, or at least about 3.2 kDa, or at least about 3.6 kDa, or at least about 4.0 kDa, or at least about 4.4 kDa, or at least about 4.8 kDa or at least about 5.2 kDa, or at least 5.6 kDa, or at least 6.0 kDa, or at least 6.4 kDa or more. In some embodiments, the coatings can have PPO segments with molecular weights of from about 1.8 kDa to about 10 kDa, or from about 2 kDa to about 5 kDa, or from about

[0206] 2.5 kDa to about 4.5 kDa, or from about 2.5 kDa to about 3.5 kDa, or from about 3.0 kDa to about 5.0 kDa, or from about 3.0 kDa to about 6.0 kDa, or from about 4 kDa to about 6 kDa, or from 4.0 kDa to about 7.0 kDa. In some embodiments, at least about 10%, or at least about 25%, or at least about 50%, or at least about 75%, or at least about 90%, or at least about 95%, or at least about 99% or more of the hydrophobic regions in these coatings have molecular weights within these ranges. In some embodiments, the coatings are biologically inert. Compounds that generate both a hydrophilic surface and an uncharged or substantially neutrally-charged surface can be biologically inert.

[0207] Representative coatings that may be used with the conjugate compounds disclosed herein can include molecules having, for example, hydrophobic segments such as PEG segments with molecular weights of at least about 1 .8 kDa, or at least about 2 kDa, or at least about 2.4 kDa, or at least about 2.8 kDa, or at least about 3.2 kDa, or at least about

[0208] 3.6 kDa, or at least about 4.0 kDa, or at least about 4.4 kDa, or at least about 4.8 kDa, or at least about 5.2 kDa, or at least 5.6 kDa, or at least 6.0 kDa, or at least 6.4 kDa or more. In some embodiments, the coatings can have PEG segments with molecular weights of from about 1.8 kDa to about 10 kDa, or from about 2 kDa to about 5 kDa, or from about 2.5 kDa to about 4.5 kDa, or from about 2.5 kDa to about 3.5 kDa. In some embodiments, at least about 10%, or at least about 25%, or at least about 50%, or at least about 75%, or at least about 90%, or at least about 95%, or at least about 99% or more of the hydrophobic regions in these coatings have molecular weights within these ranges. In some embodiments, the coatings are biologically inert. Compounds that generate both a hydrophilic surface and an uncharged or substantially neutrally-charged surface can be biologically inert.

[0209] Representative coatings that may be used with the conjugate compounds disclosed herein can include molecules having, for example, segments such as PLGA segments with molecular weights of at least about 4 kDa, or at least about 8 kDa, or at least about 12 kDa, or at least about 16 kDa, or at least about 20 kDa, or at least about 24 kDa, or at least about 28 kDa, or at least about 32 kDa, or at least about 36 kDa, or at least about 40 kDa, or at least about 44 kDa, of at least about 48 kDa, or at least about 52 kDa, or at least about 56 kDa, or at least about 60 kDa, or at least about 64 kDa, or at least about 68 kDa, or at least about 72 kDa, or at least about 76 kDa, or at least about 80 kDa, or at least about 84 kDa, or at least about 88 kDa or more. In some embodiments, at least about 10%, or at least about 25%, or at least about 50%, or at least about 75%, or at least about 90%, or at least about 95%, or at least about 99% or more of the regions in these coatings have molecular weights within these ranges. In some embodiments, the coatings are biologically inert. Compounds that generate both a hydrophilic surface and an uncharged or substantially neutrally-charged surface can be biologically inert.

[0210] The conjugate compound disclosed herein are in embodiments non-toxic and / or pharmaceutically acceptable and are therefore suitable for in vivo use in treating diseases or disorders such as cancer, or for diagnostic or imaging purposes. Compositions comprising the conjugate compound disclosed herein are also contemplated, including pharmaceutically acceptable compositions (e.g. formulations). Various types of pharmaceutical compositions can be used, depending on the desired form of administration. For example, aqueous compositions comprise an effective amount of the metal nanorods disclosed herein dissolved and / or dispersed in a pharmaceutically acceptable carrier and / or aqueous medium. The pharmaceutical compositions disclosed herein can further comprise supplementary active ingredients, such as an anti-cancer agent.

[0211] Other delivery methods of the present invention comprise compositions comprising one or more lipids associated with the conjugate compound. A lipid is a substance that is characteristically insoluble in water and extractable with an organic solvent. Lipids include, for example, the substances comprising the fatty droplets that naturally occur in the cytoplasm as well as the class of compounds which are well known to those of skill in the art which contain long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, amino alcohols, and aldehydes. These examples are not meant to be limiting, and compounds other than those specifically disclosed herein that are understood by one of skill in the art as lipids are also encompassed by the compositions and methods disclosed herein. For example, a lipid may be naturally occurring or synthetic (i.e., designed or produced by man). However, a lipid is usually a biological substance. Biological lipids are well known, and include for example, neutral fats, phospholipids, phosphoglycerides, steroids, terpenes, lysolipids, glycosphingolipids, glycolipids, sulphatides, lipids with ether and ester-linked fatty acids and polymerizable lipids, and combinations thereof. In particular embodiments, a lipid comprises a liposome. A liposome is a generic term encompassing a variety of single and multilamellar lipid vehicles formed by the generation of enclosed lipid bilayers or aggregates. Liposomes may be characterized as having vesicular structures with a bilayer membrane, generally comprising a phospholipid, and an inner medium that generally comprises an aqueous composition. A multilamellar liposome has multiple lipid layers separated by aqueous medium. They form spontaneously when lipids comprising phospholipids are suspended in an excess of aqueous solution. The lipid components undergo self-rearrangement before the formation of closed structures, entrapping water and dissolved solutes between the lipid bilayers. Lipophilic molecules or molecules with lipophilic regions may also dissolve in or associate with the lipid bilayer.

[0212] In particular embodiments, a conjugate compound may be, for example, encapsulated in the aqueous interior of a liposome, interspersed within the lipid bilayer of a liposome, attached to a liposome via a linking molecule that is associated with both the liposome and the metal nanorod, entrapped in a liposome, complexed with a liposome, etc. A liposome used as disclosed herein may be made by different methods, as would be known to one of ordinary skill in the art. Phospholipids can form a variety of structures other than liposomes when dispersed in water, depending on the molar ratio of lipid to water. At low ratios the liposome is the typical structure. The size of a liposome varies depending on the method of synthesis. Liposomes disclosed herein can have a variety of sizes. In certain embodiments, the liposomes are small, e.g., less than about 100 nm, about 90 nm, about 80 nm, about 70 nm, about 60 nm, or less than about 50 nm in external diameter. In preparing such liposomes, any protocol disclosed herein, or as would be known to one of ordinary skill in the art may be used. Additional non-limiting examples of preparing liposomes are described in U.S. Patent Nos. 4,728,575, 4,737,323, 4,533,254, 4,162,282, 4,310,505, and 4,921 ,706; A comprehensive review of lipid vesicles and methods for their preparation are described in "Liposome Technology" (1984. Gregoriadis G. ed. CRC Press Inc Boca Raton Florida Vol I II & III).

[0213] Liposomes interact with cells to deliver agents via four different mechanisms: Endocytosis by phagocytic cells of the reticuloendothelial system such as macrophages and / or neutrophils; adsorption to the cell surface, either by nonspecific weak hydrophobic and / or electrostatic forces, and / or by specific interactions with cell-surface components; fusion with the plasma cell membrane by insertion of the lipid bilayer of the liposome into the plasma membrane, with simultaneous release of liposomal contents into the cytoplasm; and / or by transfer of liposomal lipids to cellular and / or subcellular membranes, and / or vice versa, without any association of the liposome contents. Varying the liposome formulation can alter which mechanism is operative, although more than one may operate at the same time.

[0214] A skilled person realizes that the conjugate compounds and methods disclosed herein can be employed in a variety of types of experimental, therapeutic and diagnostic procedures, including in vitro or in vivo procedures. In another embodiment, systems, devices, materials, and techniques are described for minimally invasive active targeting, fluorescent imaging, and NIR photothermal treatment of tumours, for example, which can be applied to a variety of cancer types.

[0215] II. Methods of Making Conjugate Compounds

[0216] Various permutations of components can be used to make the conjugate compound disclosed herein. X1and X2are the same or different, and are each independently selected from a non-covalent linker or a covalent linker and may be cleavable. In other embodiments, at least one of X1and X2is a cleavable linker. In examples, either X1or X2is a cleavable linker such that either X1or X2is cleaved to release the TpA or the TTA, typically, release the TpA.

[0217] In an embodiment, the method for making the conjugate compound disclosed herein is provided. The method comprises combining TTA-CRG1 and CRG2-MNP-X2-TpA to make TTA-X1-MNP-X2-TpA, wherein CRG is a conjugate reactive group. The method may further comprise combining MNP-X2-TpA and CRG2 to make the CRG2-MNP-X2-TpA.

[0218] In an embodiment, the method for making the conjugate compound disclosed herein is provided. The method comprises combining TTA-X1-MNP-CRG3 and CRG4-TpA to make TTA-X1-MNP-X2-TpA, wherein CRG3 and CRG4 are different, and are each selected to form X2, optionally, CRG4 is a portion of the TpA, wherein CRG is a conjugate reactive group. The method may further comprise combining TTA-X1-MNP and CRG3 to make the TTA-X1-MNP-CRG3.

[0219] In an embodiment, the method for making the conjugate compound disclosed herein is provided. The method comprises combining TTA-CRG1 , CRG2-MNP-CRG3, and CRG4-TpA to make TTA-X1-MNP-X2-TpA, wherein CRG is a conjugate reactive group. The method may further comprise combining TTA-CRG1 , CRG2-MNP-CRG3, and CRG4-TpA, in any order. In embodiments, the method further comprises combining TTA-X1-MNP and CRG3 to make TTA-X1-MNP-CRG1.

[0220] Some specific method embodiments include the following: i) Embodiment A for making TTA-X1-MNP- X2-TpA: a) TTA-CRG1

[0221] CRG1 can be any suitable CRG disclosed herein. CRG1 is combined with TTA to form CRG1-TTA or is already an integral part of the TTA. The TTA can be any suitable TTA disclosed herein. b) CRG2-MNP

[0222] CRG2 can be any suitable CRG disclosed herein. CRG2 is combined with MNP to form CRG2-MNP or is already an integral part of the MNP. The MNP can be any suitable MNP disclosed herein. c) TTA-X1-MNP

[0223] X1can be a non-covalent or covalent linker formed from the combination of TTA- CRG1 of a) and CRG2-MNP of b), wherein CRG1 and CRG2 react to form a covalent linker (e.g. amide) or combine to form a non-covalent linker (e.g. DNA) disclosed herein.

[0224] X1can be a non-covalent linker formed from the direct non-covalent bonding of TTA and MNP (TTA-MNP). d) TTA-X1-MNP-CRG3

[0225] TTA-X1-MNP, as described in c), is combined with CRG3 to form TTA-X1-MNP- CRG3. CRG3 can be any suitable CRG disclosed herein. e) CRG4-TpA

[0226] CRG4 can be any suitable CRG disclosed herein. CRG4 is combined with TpA to form CRG4-T pA or is already an integral part of the T pA. The T pA can be any suitable T pA disclosed herein. f) TTA-X1-MNP- X2-TpA

[0227] TTA-X1-MNP-CRG3 of d) is combined with CRG4-TpA of e) to form TTA-X1-MNP- X2-TpA. ii) Embodiment B for making TTA-X1-MNP- X2-TpA: a) CRG4-TPA

[0228] CRG4 can be any suitable CRG disclosed herein. CRG4 is combined with TpA to form CRG4-TpA or is already an integral part of the TpA. The TpA can be any suitable TpA disclosed herein. b) MNP-CRG3

[0229] CRG3 can be any suitable CRG disclosed herein. CRG3 is combined with MNP to form MNP-CRG3 or is already an integral part of the MNP. The MNP can be any suitable MNP disclosed herein. c) MNP-X2-TpA X2can be a non-covalent or covalent linker formed from the combination of CRG4- T pA of a) and MNP-CRG3 of b), wherein CRG3 and CRG4 react to form a covalent linker (e.g. amide) or combine to form a non-covalent linker (e.g. DNA) disclosed herein. CRG4 and CRG4 can be any suitable CRG disclosed herein.

[0230] X2can be a non-covalent linker formed from the direct non-covalent bonding of MNP and TpA (MNP-TpA). d) CRG2-MNP-X2-TpA

[0231] MNP-X2-TpA, as described in c), is combined with CRG2 to form CRG2-MNP-X2- TpA. CRG2 can be any suitable CRG disclosed herein. e) TTA-CRG1

[0232] CRG1 can be any suitable CRG disclosed herein. CRG1 is combined with TTA to form TTA-CRG1 or is already an integral part of the TTA. The TTA can be any suitable TTA disclosed herein. f) TTA-X1-MNP- X2-TpA

[0233] CRG2-MNP-X2-TpA of d) is combined with TTA-CRG1 to form TTA-X1-MNP- X2- TpA. iii) Embodiment C for making TTA-X1-MNP- X2-TpA: a) CRG2-MNP-CRG3

[0234] MNP is combined with CRG1 and CRG3 (e.g. stepwise (in any order) or together), which each can be any suitable CRG disclosed herein, to form CRG2-MNP-CRG3. b) TTA-CRG1

[0235] CRG1 can be any suitable CRG disclosed herein. CRG1 is combined with TTA to form TTA-CRG1 or is already an integral part of the TTA. c) CRG4-TPA

[0236] CRG4 can be any suitable CRG disclosed herein. CRG4 is combined with TpA to form CRG4-TpA or is already an integral part of the TpA. The TpA can be any suitable TpA disclosed herein. d) TTA-X1-MNP- X2-TpA

[0237] CRG2-MNP-CRG3 of a) is combined with TTA-CRG1 of b) and CRG4-TpA of c) (e.g. stepwise (in any order) or together) to form TTA-X1-MNP- X2-TpA.

[0238] Other specific method embodiments are shown in Figures 1A-1 E. In typical embodiments, the MNP is GNR.

[0239] With respect to the methods herein, it is understood that the components of making the conjugate compound disclosed herein, may be in the form of dispersions, such as colloidal dispersions. In embodiments, DNA linkers can be selected for X1and X2as one or two sets / pair of single strand DNA (ssDNA). Within each set, one strand of ssDNA is complementary to the other ssDNA, however, two ssDNA from two different sets are not complementary to each other. Complementary ssDNA that are binding to GNRs can have a thiol (-SH) or disulfide (-S-S-) functional group either on 3’ or 5’ end. Complementary ssDNA for T pA and TTA can be functionalized according to the requirements of conjugation chemistry. The number of base pairs for all these ssDNA can be any suitable base pairs per strand (e.g. 2 to 30 base pairs).

[0240] Examples include linkers resulting from EDC-NHS chemistry (labile or non-labile linker) or two complementary single strands of DNA (heat labile linker). One strand of ssDNA(1 ’) of a first set of a complementary DNA pair (DNA(1)) is coupled to MNP (Conjugate 1) and the other ssDNA(1”) of the first set of the complementary DNA pair (DNA(1)) is coupled to TTA (Conjugate 2). Conjugates 1 and 2 are combined such that the ssDNA(1 ’) and ssDNA(1”) bind to form the complementary DNA pair (DNA(1)); TTA- DNA(1)-MNP (Conjugate 3). The Conjugate 3 is further conjugated to a single strand DNA(2’) from a second set of complementary DNA pair (DNA(2)) to form TTA-DNA(1)- MNP-ssDNA(2’) (Conjugate 4). The other ssDNA(2”) from the second set of complementary DNA pair (DNA(2)) is coupled to TpA (Conjugate 5). Conjugates 4 and 5 are combined such that the ssDNA(2’) and ssDNA(2”) bind to form the complementary DNA pair (DNA(2)); TTA-DNA(1)-MNP-DNA(2)-TpA (Conjugate 6).

[0241] It is noted that in certain embodiments, ssDNA can be used for delivery of MNPs and TpA at the same tumour sites (e.g. both delivered to either extracellular or intracellular) or delivery of MNP and TpA at different tumour sites (extracellular or intracellular).

[0242] In various embodiments, one of the following may be used to make the conjugate compound (TTA-X1-MNP-X2-TpA). i) A pair of ssDNA is used as a cleavable linker to conjugate TTA to MNPs (TTA-X1- MNP, X1is dsDNA(1)). An ssDNA(1 ’) of a pair of ssDNA is conjugated to the TTA (TTA- ssDNA(1 ’)) and the ssDNA(1”) of the pair of ssDNA is conjugated to the MNP (ssDNA(1”)-MNP). TTA- ssDNA(1’) and ssDNA(1”)-MNP are combined to make TTA-X1-MNP, wherein X1is DNA(1).

[0243] An ssDNA(2’) of a second pair of ssDNA is conjugated to the MNP of TTA-X1-MNP (TTA-X1-MNP-ssDNA(2’)). The TTA-X1-MNP-ssDNA(2’) is combined with ssDNA(2”)-TpA, which is the complementary ssDNA(2”) of the second pair of ssDNA, to make TTA-X1-MNP-X2-TpA, wherein X2is DNA(2). ii) A non-DNA linker (e.g. can include a functional group linker) to conjugate TTA to MNPs ((TTA-X1-MNP, X1includes a functional group linker or is a non-covalent linker). TTA-CRG1 and CRG2-MNP are combined to make TTA-X1-MNP, wherein CRG1 (Conjugate Reactive Group) and CRG2 are different, and are each selected to form X1.

[0244] An ssDNA(1 ’) of a pair of ssDNA is conjugated to the MNP of TTA-X1-MNP to make TTA-X1-MNP-ssDNA(1 ’). The TTA-X1-MNP-ssDNA(1 ’) is combined with ssDNA(1”)- T pA, which is the complementary ssDNA(1 ”) of the pair of ssDNA, to make TTA-X1- MNP-X2-TpA, wherein X2is DNA(1). iii) A pair of ssDNA is used as a cleavable linker to conjugate TTA to MNPs (TTA-X1- MNP, X1is dsDNA(1)). An ssDNA(1 ’) of a pair of ssDNA is conjugated to the TTA (TTA- ssDNA(1 ’)) and the ssDNA(1”) of the pair of ssDNA is conjugated to the MNP (ssDNA(1”)-MNP). TTA- ssDNA(1’) and ssDNA(1”)-MNP are combined to make TTA-X1-MNP, wherein X1is DNA(1).

[0245] A non-DNA linker (e.g. can include a functional group linker) to conjugate TpA to MNP of TTA-X1-MNP. The MNP of TTA-X1-MNP forms TTA-X1-MNP-CRG1 , which is combined with CRG2-TpA, to make TTA-X1-MNP-X2-TpA, wherein X2is includes a functional group linker or is a non-covalent linker. CRG1 (Conjugate Reactive Group) and CRG2 are different, and are each selected to form X2.

[0246] Protecting TTA-X1-MNP-X2-TpA

[0247] Protecting the conjugate compound upon exposure to various elements, such as whole blood, are described herein. Polymers can be used to protect the conjugate compound, such as synthetic polymers and / or natural polymers. Synthetic polymers are those suitable for use in mammals. Examples of synthetic polymers include PEG (polyethylene glycol), PVP (polyvinylpyrrolidone), poly lactic-co-glycolic acid (PLGA), PSS (polystyrene sulfonate), PNIPAM (poly(N-isopropylacrylamide)), ploxamer, diblock or triblock polymers, pH or thermo-responsive polymers, or a combination thereof. Natural polymers are those suitable for use in mammals. Examples of natural polymers include glycoproteins, albumin, gelatine, collagen, or a combination thereof. In addition to polymer(s) or separately, phospholipids (both low and high density), micelles, vesicles, liposomes, solid lipid nanoparticle (SLN), nanostructured lipid carriers (NLC), or a combination thereof. For the protection of the conjugate compound with polymer(s), a selected polymer may have a thiol or disulphide functional group (e.g. at a terminal end) or has been modified to include this functional group. Nano-aggregates such as micelles, vesicles, liposomes, solid lipid nanoparticle (SLN), nanostructured lipid carriers (NLC) and / or supramolecular vesicles are described for encapsulation and / or surface adsorption of the conjugate compound for intratumoral injection.

[0248] According to certain embodiments, the pharmaceutical composition is formulated for parenteral administration, e.g., formulated for injection via the intravenous, intramuscular, sub-cutaneous, intralesional, and intraperitoneal routes. Typically, such compositions are prepared either as liquid solutions or suspensions; solid forms suitable for using to prepare solutions and / or suspensions upon the addition of a liquid prior to injection can also be prepared; and the preparations can also be emulsified. The conjugate compound compositions disclosed herein can be formulated into a composition in a neutral and / or salt form for example. Any pharmaceutically acceptable salt known to a person skilled in the art can be used, providing it would not interfere with the function of the metal nanorods.

[0249] Sterile injectable solutions are generally prepared by incorporating the active compounds, specifically the conjugate compound in the required amount in the appropriate solvent with other ingredients, as required, followed by filtered sterilization.

[0250] Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and / or the required other ingredients as disclosed herein above. In the case of sterile powders for the preparation of sterile injectable solutions, the typical methods of preparation are vacuumdrying and / or freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. The preparation of more, and / or highly, concentrated solutions for direct injection is also contemplated, where the use of DMSO as solvent is envisioned to result in extremely rapid penetration, delivering high concentrations of the active agents to a small target area.

[0251] In another method, the conjugate compound may be wrapped in a polymer. In embodiments, the wrapping polymers may include proteins, gelatin, bovine serum albumin, polystyrene sulfonate, polyethylene oxides, thiolated polyethylene oxides, thiolated polyethyene oxides with terminating carboxylic acid functionalities, thiolated polyethyene oxides with terminating amine acid functionalities, and combinations thereof.

[0252] In an embodiment, the wrapping polymer may form covalent and / or non-covalent bonds with other polymers, proteins, etc. In certain embodiments, the carboxylic ending of thiolated polyethylene oxides is bound to protein(s), polypeptide(s), antibodie(s), antibody fragment(s), IgG class of antibody, a polyclonal antibody, a monoclonal antibody, and combinations thereof. In another embodiment, the amine ending thiolated polyethylene oxide is bound to protein(s), polypeptide(s), antibodie(s), antibody fragment(s), IgG class of antibody, a polyclonal antibody, a monoclonal antibody, and combinations thereof. Such covalently bounded bioconjugates may be formed, for example, from any metal nanorods disclosed herein, a polymer, and, for example, an antibody, protein(s), polypeptide(s), antibodie(s), antibody fragment(s), IgG class of antibody, a polyclonal antibody, a monoclonal antibody. In an embodiment, single or double stranded nucleic acid may be tethered to MNPs with metal-thiol bonds. In an embodiment, an oligonucleotide may be tethered to metal nanorods with metal-thiol bonds.

[0253] In embodiments, the wrapping polymer is covalently bonded to the MNP and can form a non-covalent bond with TTA and / or TpA.

[0254] The conjugate compound can comprise a variety of capping agents are described. In an embodiment, methods of capping the conjugate compound with a non-surfactant involve first removal of a solvent or excess surfactant from the metal nanorod solution followed by addition of an aqueous solution of capping agent(s). In typical embodiments, about 95% to about 98% of the solvent is removed and a similar quantity of the aqueous solution of the new capping agent(s) is added. In a more specific embodiment, the method comprises removal of about 95% to about 98% of solvent from the metal nanorod solution, followed by the addition of a similar amount of an aqueous solution of a first capping agent (e.g. an ionic polymer, typically an anionic polymer), and allowing the solution to equilibrate for a period of time. For example, the mixture may be equilibrated to a temperature of about 4°C to about 25°C for at least about 1 hour. Then removing about 95% to about 98% of resultant solvent from the resultant nanoparticle pellets, for example, by using a centrifugal method, and additional dispersion of the resultant conjugate compound into an aqueous solution of a second capping agent (e.g. same or different from first capping agent).

[0255] In embodiments, the capping agent of conjugate compound in a colloidal solution form may be a mixture of surfactant and a thiolated polymer (polyethylene glycol of mwt. of about 500Da to about 50kDa) or it can be poly lactic-co-glycolic acid (PLGA) of mwt. of about 500Da to about 50kDa. Other ranges include for example, about 600Da to about 50kDa, about 700Da to about 50kDa, about 800Da to about 50kDa, about 900Da to about 50kDa, about 1 kDa to about 50kDa, about 1 kDa to about 40kDa, about 1 kDa to about 30kDa, about 1 kDa to about 20kDa, or about 1 kDa to about 10kDa. In an embodiment, the capping agent of conjugate compound in a colloidal solution form may be a mixture of surfactant from a surfactant solution and a thiolated polymer (polyethylene glycol of mwt. of about 500Da to about 50kDa, Other ranges include for example, about 600Da to about 50kDa, about 700Da to about 50kDa, about 800Da to about 50kDa, about 900Da to about 50kDa, about 1 kDa to about 50kDa, about 1 kDa to about 40kDa, about 1 kDa to about 30kDa, about 1 kDa to about 20kDa, or about 1 kDa to about 10kDa). In an embodiment, the capping agent of conjugate compound may be a mixture of surfactant, a co-surfactant, and small biomolecules. The small biomolecules may be selected from a general class of flavonoids, antioxidants, aromatic acids, amino acids, monohydroxybenzoic acid, monosaccharides, disaccharides, bile salt, nucleotides, or combinations thereof. In an embodiment, co-capping agent(s) may be added such as quercetin, epigallocatechin gallate, curcumin, glutathione, ascorbic acid, citric acid, anthranilic acid, cinnamic acid, bile acid, and p-hydroxybenzoic acid, metal anionic salts of biological acid(s), or combinations thereof.

[0256] III. Conjugate Compounds: Methods of Treatment and Uses Thereof

[0257] In embodiments, the conjugate compound disclosed herein can be used to treat cancer. The method can comprise administering a therapeutically effective amount of the conjugate compound disclosed herein, or a pharmaceutical composition thereof, to a subject in need thereof for treating cancer. In other embodiments, the use of the conjugate compound or the pharmaceutical composition thereof for treatment of cancer in a subject is provided. The methods and uses of the conjugate compound disclosed herein or the pharmaceutical composition thereof may improve therapeutic outcomes while minimizing side-effects in a subject, as further described below.

[0258] In specific embodiments, the conjugate compound disclosed herein can be used to treat a tumour. The method can comprise administering a therapeutically effective amount of the conjugate compound disclosed herein, or a pharmaceutical composition thereof, to a subject in need thereof for treating a tumour. In other embodiments, the use of the conjugate compound or the pharmaceutical composition thereof for treatment of a tumour in a subject is provided. The methods and uses of the conjugate compound disclosed herein or the pharmaceutical composition thereof may improve therapeutic outcomes while minimizing side-effects in a subject, as further described below.

[0259] Therefore, cancer may be treatable and / or preventable (e.g. prevention of further cancer development and / or metastasis) by administration or delivery of the conjugate compound as disclosed herein, or a pharmaceutical composition thereof.

[0260] In embodiments, the method comprises targeting a cancer cell with the conjugate compound or the pharmaceutical composition thereof. In embodiments, the TTA of the conjugate compound binds to the cancer cell, and in other embodiments, the TTA of the conjugate compound binds to the tumour (e.g. tumour cells including cancer cells). The TTA is the same as described above, so the same description thereof is applicable here and will not be repeated for the sake of brevity. The TTA can function such that the conjugate compound can reach its intended target (e.g. cancer cells) with the TpA. As a result, in embodiments, since the TpA is carried to the tumour or the cancer cells, the TTA can function to reduce toxicity by reducing off-target delivery of the TpA. The TpA is the same as described above, so the same description thereof is applicable here and will not be repeated for the sake of brevity.

[0261] Thus, the conjugate compound disclosed herein may be administered to a cell (e.g. cancerous cells) or tissue (e.g. tumour) using targeting schemes involving specific chemical interactions (e.g., antigen-antibody binding, etc.), such as for example, through the use of the TTA described above so as to directly target the conjugate compound to the tumour, or may consist of the delivery of the conjugate compound to the desired area (e.g. tumours). The conjugate compound disclosed herein may be delivered in the form of a pharmaceutical composition. The targeting may be to the surface of the subject cells and / or tissue, such as to the surface of the tumour(s), or it may be to other, interior sites, as such, for example, internalization of the conjugate compound into to the tumour for treatment thereof. In this way, since the methods and uses disclosed herein can involve direct targeting of the conjugate compound to the cancer cells / tumour cells, lower dosages of the TpA may be used and / or unwanted off-target side-effects of the TpA may also be minimized since the TpA is directed to the tissue to be treated, as opposed to, healthy tissue (e.g. released systemically).

[0262] In embodiments, the method further comprises cleaving X1or X2of the conjugate compound disclosed herein. As described in greater detail above, cleavage of one or both of the linkers of the conjugate compound may be through the use of stimulus, so that one or more of the components (TpA, MNP, and TTA) of the conjugate compound separate. The stimulus includes, but is not limited to, photo-irradiation, such as irradiation at certain wavelength(s) to control cleavage (e.g. UV, visible, or NIR), enzymes, nucleophilic / basic reagents, reducing agents, electrophilic / acidic reagents, organometallic and metal reagents, oxidizing reagents, or a combination thereof. Thus, in embodiments, the method or use comprises applying stimulus to the conjugate compound to cause cleavage of one or both cleavable linkers. In embodiments, the stimulus comprises pH, enzymes, or EMR. The type of stimulus selected to achieve cleavage will depend on the cleavable linker. One skilled in the art would understand the appropriate stimulus suitable for cleavage of the selected linkers of the conjugate compound. For example, if a specific peptide linker is used, then an enzyme specific for cleaving the peptide bonds of that linker would be employed. In typical embodiments, the stimulus comprises application of EMR. In embodiments, the conjugate compound can be used in conjunction with other cancer treatments, for example, and without being limited thereto, PTT.

[0263] The EMR can be any EMR, such as, for example, radiowaves, microwaves, infrared, near infrared, visible light, ultraviolet light, x-rays, gamma radiation, or a combination thereof. In typical embodiments, the EMR is near infrared EMR. "Near infrared EMR" or "NIR-EMR", as used herein, refers to electromagnetic radiation having wavelengths in a range of about 650 nm to about 3000 nm, about 650 nm to about 2500 nm, about 650 nm to about 1100 nm; about 700 nm to about 1100 nm; about 750 nm to about 1100 nm; about 800 nm to about 1100 nm; about 850 nm to about 1100 nm; about 700 nm to about 1000 nm; about 750 nm to about 950 nm; about 800 nm to about 1000 nm; about 850 nm to about 1000 nm or about 800 nm to about 850 nm, inclusive of all endpoints and subintervals of the aforementioned ranges.

[0264] The conjugate compound disclosed herein has optical properties, in that it can absorb light in the NIR of about 700 nm to about 1000 nm and can be easily detected when a laser beam with a similar wavelength (approximately 800 nm) irradiated the compound. Laser light at this wavelength is typically used, as a high percentage of this wavelength of light can pass through biological tissue . As a result, the conjugate compound can be injected directly into the bloodstream and their location / distribution can be determined using a safe laser light, also referred to as infrared irradiation. In this way, the methods and uses disclosed herein using EMR-NIR can also increase the subject’s level of comfort during the treatment thereof.

[0265] In embodiments, the EMR is used to irradiate the conjugate compound or composition(s) disclosed herein. In embodiments, the MNP of the conjugate compound absorbs the EMR and generates heat to temperatures of about 40°C or greater, such as, for example, in a range of about 42 °C to about 45 °C, and more particularly about 44 °C, which heats the target tissue (e.g. the tumour of the subject) to similar temperatures to inhibit growth / reduce the tumour.

[0266] The EMR may be generated by, for example, a light-emitting diode (LED) or a laser generator. LEDs and laser generators that are capable of generating EMR, and more particularly NIR-EMR, are available. As non-limiting examples, such laser generators may include semiconductor laser generators (also referred to as laser diodes) with a nonlimiting example being a vertical-cavity surface-emitting laser (VCSEL), and solid-state laser generators (i.e., a laser that uses a solid gain medium) with a non-limiting example being a titanium sapphire laser generator. Devices capable of delivery EMR are those described in US63 / 562461 , the entirety of which is incorporated by reference.

[0267] In embodiments, the EMR (e.g. NIR) functions to at least one of cleave X1, cleave X2, and excite the MNP of the conjugate compound. In certain embodiments, the EMR functions to excite the MNP to generate heat. In further embodiments, the EMR functions to cleave X2to release the therapeutic agent. In embodiments, the conjugate compound or composition(s) generate heat to inhibit / reduce growth of tumour tissue. To this regard, the conjugate compound disclosed herein can heat-up when irradiate. This property results in their ability to increase the temperature locally, for example in the immediate vicinity of a specific target. Thus, when the target is a tumour or individual cancer cells, the tumour or cell will be damaged or destroyed. This allows for non-invasive anti-cancer therapy using the conjugate compound disclosed herein. Moreover, with the use of the TTA as disclosed herein, the treatment is targeted since the conjugate compound is delivered primarily to the cancer cell or the tumour.

[0268] The conjugate compound can therefore assist in at least two types of treatment, simultaneously or synchronously, comprising releasing the TpA upon exposure to stimulus (e.g. EMR), along with hyperthermia via the MNP. For example, depending on the linkers used in the conjugate compound, the TpA may be released from the conjugate compound after the conjugate compound is exposed to the stimulus such that a synchronous treatment schedule of hyperthermia, followed by immunotherapy / chemotherapy via the TpA is possible. In another example, the TpA may be released simultaneously with the exposure of the conjugate compound to the stimulus such that phototherapy and the immunotherapy treatments occur simultaneously.

[0269] In other embodiments, the conjugate compound (TTA-X1-MNP-X2-TpA) provides at least more benefits, such as an additive or synergistic effect. This effect is in comparison to: a) stepwise delivery of 1) MNPs with PTT; and 2) TpAs; b) stepwise delivery of 1) TpAs; and 2) MNPs with PTT; c) a composition of MNPs and TpAs with PTT; d) stepwise delivery of 1) TTA-MNPs with PTT; and 2) TpAs; e) stepwise delivery of 1) TpAs; and 2) TTA-MNPs with PTT; or f) a composition of TTA-MNPs and TpAs.

[0270] Unlike the conjugate compound disclosed herein, each of a) to f) would not reduce the side-effects of off-target delivery of the T pA since none of a) to f) allow for targeting of the therapeutic agent to the cancer (e.g. tumour). In addition, the synergistic effect (e.g. more than additive effect) provided by targeting the conjugate compound to the cancer cell and / or tumour can include, for example, lower dosages of one or more of T pA, TTA, and MNPs to achieve the same or better treatment; same dosages but more effective treatment; less off-target delivery and / or actions of therapeutic agents; less toxicity; etc.

[0271] The cancer disclosed herein may be any cancer. A subject that has a cancer or a tumour is a subject that can have objectively measurable cancer cells present. In some embodiments, a tumour may be or comprise cells that are precancerous (e.g., benign), malignant, pre-metastatic, metastatic, and / or non-metastatic. In some embodiments, the cancer may be characterized by a solid tumour, and in other embodiments, the relevant cancer may be characterized by a hematologic tumour. Examples of different types of cancers include, for example, hematopoietic cancers including leukemias, lymphomas (Hodgkin's and non-Hodgkin's), myelomas and myeloproliferative disorders; sarcomas, melanomas, adenomas, carcinomas of solid tissue, squamous cell carcinomas mouth, throat, larynx, and lung, liver cancer, genitourinary cancers such as prostate, cervical, bladder, uterine, and endometrial cancer and renal cell carcinomas, bone cancer, pancreatic cancer, skin cancer, cutaneous or intraocular melanoma, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, head and neck cancers, breast cancer, brain cancer, kidney cancer, gastro-intestinal cancers and nervous system cancers, benign lesions such as papillomas, and the like.

[0272] In embodiments, the cancer is selected from brain tumour, nerve sheath tumour, breast cancer, colon cancer, colorectal cancer, skin cancer, lung cancer, hepatocellular cancer, gastric cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, cancer of the urinary tract, thyroid cancer, renal cancer, renal cell carcinoma, carcinoma, melanoma, lymphoma, head and neck cancer, brain cancer, and prostate cancer, including but not limited to androgen-dependent prostate cancer and androgen-independent prostate cancer. In embodiments, the cancer is prostate cancer, and in other embodiments, the cancer is breast cancer.

[0273] The tumour tissue and / or tumour to be treated may be of any source. In embodiments, the tumour tissue or the tumour, is derived from a peripheral tumour, breast cancer, colon cancer, colorectal cancer, skin cancer, lung cancer, hepatocellular cancer, gastric cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, cancer of the urinary tract, thyroid cancer, renal cancer, renal cell carcinoma, carcinoma, melanoma, lymphoma, head and neck cancer, brain cancer, and prostate cancer, including but not limited to androgen-dependent prostate cancer and androgenindependent prostate cancer. In embodiments, the tumour is derived from prostate cancer, and in other embodiments, the tumour is derived from breast cancer.

[0274] The subject referred to herein is, typically, a mammal, and most typically a human that that is suffering from cancer and / or has a tumour that can be treated by the methods and uses disclosed herein. Thus, the subject has detectable cancer cells which can be treated by the methods or uses disclosed herein. The methods and uses disclosed herein may also have a preventative function, in that, if cancer cells are detected in the subject, and the subject is subsequently treated by the methods and uses disclosed herein, further development of cancer, or metastasis may be prevented through application of the methods and uses disclosed herein. Thus, the uses and methods disclosed herein can be used to treat, such as, to destroy a tumour, if for example, the conjugate compound are provided (e.g. administered) to the subject who suffers from cancer and / or has a tumour disclosed herein. In alternative embodiments, further cancer and / or tumour development may be halted by treating the cancer and / or tumour with the methods disclosed herein. For purposes of the present disclosure, the subject may have a single cancer cell, a single cancer, a single tumour, multiple cancer cells, multiple cancers (e.g. breast and skin cancer) and / or multiple tumours, that are to be treated by the uses and methods disclosed herein.

[0275] In some embodiments, treatment refers to an increased survival (e.g. an increased survival time). For example, treatment can result in an increased life expectancy of a subject. In some embodiments, treatment with the conjugate compound results in an increased life expectancy of a subject by more than about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 105%, about 110%, about 115%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, about 175%, about 180%, about 185%, about 190%, about 195%, about 200% or more, as compared to the average life expectancy of one or more control individuals with similar disease without treatment. In some embodiments, treatment according to the present invention results in an increased life expectancy of a patient (subject) by more than about 6 month, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, about 10 years or more, as compared to the average life expectancy of one or more control individuals with a similar disease without treatment. In some embodiments, treatment results in long term survival of a patient. As used herein, the term “long term survival” refers to a survival time or life expectancy longer than about 20 years, 30 years, 40 years, 50 years, 60 years, or longer.

[0276] Administration "in combination with" one or more further therapeutic agents can include simultaneous (concurrent) and consecutive administration in any order.

[0277] In embodiments, a therapeutically effective amount of at least one conjugate compound may be used. Administration may comprise a single administration, or alternatively, comprise a series of administrations or systematic administration of conjugate compounds with different TpA and MNPs or systematic administration of conjugate compounds with different TpAs and GNRs. The length of the treatment period can depend on factors, such as the severity of the disease, the age of the subject, the conjugate compound, the dosage of the conjugate compound, stage of cancer stage, frequency of administration, intensity of light, duration of light, a combination thereof. It will also be appreciated that the effective dosage of the agent used for the treatment may increase or decrease over the course of a particular treatment regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. The conjugate compound disclosed herein may, in embodiments, be administered before, during or after treatment with conventional therapies for the disease or disorder in question, such as cancer.

[0278] In other embodiments, the conjugate compound disclosed herein can be used in the manufacture of a medicament, and typically the medicament is for the prevention (e.g. further cancer development and / or metastasis) and / or treatment of the cancers disclosed herein. In typical embodiments, the conjugate compound, as a medicament, are for administration to a subject (e.g. mammals, typically humans) in need thereof.

[0279] The conjugate compound can be administered to mammals, typically humans. When administered as a pharmaceutical composition, the conjugate compound may be provided in combination with pharmaceutically acceptable carriers or diluents, optionally with pharmaceutically acceptable adjuvants, such as alum. The conjugate compound may, therefore, be suitably formulated into a pharmaceutical composition for administration to human subjects in a biologically compatible form suitable for administration in vivo.

[0280] Accordingly, in embodiments, the pharmaceutical composition comprises conjugate compound, in admixture with a suitable diluent or carrier. The compositions containing conjugate compound can be prepared by known methods for the preparation of pharmaceutically acceptable compositions which can be administered to subjects, such that an effective quantity of the conjugate compound is combined in a mixture with a pharmaceutically acceptable carrier. Suitable carriers are described, for example, in Remington's Pharmaceutical Sciences (2003 - 20th edition), in The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999 and in the Handbook of Pharmaceutical Additives (compiled by Michael and Irene Ash, Gower Publishing Limited, Aidershot, England (1995)), the references of which are incorporated by reference in their entirety. On this basis, the composition may include solution(s) of the conjugate compound in association with one or more pharmaceutically acceptable carrier(s) or diluent(s), and contained in buffered solution(s) with a suitable pH and iso- osmotic with the physiological fluids. Solution(s) of conjugate compound can be prepared in water suitably mixed with suitable excipients. Under ordinary conditions of storage and use, these preparations can contain a preservative to prevent the growth of microorganisms. A person skilled in the art would know how to prepare suitable formulations / compositions. In this regard, reference can be made to U.S. Patent No. 5,843,456, which is incorporated herein by reference.

[0281] The conjugate compound may be administered alone or in combination with other components / ingredients / actives. For example, the conjugate compound may be administered as a pharmaceutical composition. The described conjugate compound and / or compositions thereof, may be administered to a subject in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. The conjugate compound, and the pharmaceutical composition(s) thereof, may be administered, for example, by oral, parenteral (e.g. intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal and topical (e.g. ointment) modes of administration), buccal, sublingual, patch, pump or transdermal administration. In typical embodiments, the conjugate compound disclosed herein are administered, or are for administration, using parenteral routes of administration. It will be understood that, in aspects, the administration comprises a single administration, such as injection, for treatment rather than two or more separate administrations, such as injections. For example, the administration may comprise a single injection of treatment versus two separate injections of TTA-MNP and TpA-MNP. Likewise, the conjugate compound or composition may be formulated in a single composition, such as injection, for treatment versus two separate administrations, such as injections, of, for example, TTA- MNP and TpA-MNP.

[0282] If the conjugate compound are administered orally, the conjugate compound may be administered, for example, in the form of tablets or capsules, or as an aqueous solution or suspension. Examples may include: ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. For tablet dosage forms, depending on dose, the conjugate compound may make up from 1 wt % to 80 wt % of the dosage form, more typically from 5 wt % to 60 wt % of the dosage form. Moreover, carriers which are commonly used include lactose and corn starch, and lubricating agents, such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of magnesium stearate with sodium lauryl sulphate, are commonly added. In addition, tablets generally contain a disintegrant. Examples of disintegrants include sodium starch glycolate, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methyl cellulose, microcrystalline cellulose, lower alkyl substituted hydroxypropyl cellulose, starch, pregelatinized starch and sodium alginate. Useful diluents include lactose (monohydrate, spray dried monohydrate, anhydrous and the like), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystalline cellulose, starch and dibasic calcium phosphate dihydrate, and suitable binders include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone, pregelatinized starch, hydroxypropyl cellulose and hydroxypropyl methylcellulose. Other conventional ingredients include antioxidants, colorants, flavoring agents, preservatives and taste masking agents. Tablet blends may be compressed directly or by roller to form tablets. Tablet blends or portions of blends may alternatively be wet, dry, or melt granulated, melt congealed, or extruded before tableting. The final formulation may include one or more layers and may be coated or uncoated; or encapsulated. The formulation of tablets is discussed in detail in "Pharmaceutical Dosage Forms: Tablets, Vol. 1", by H. Lieberman and L. Lachman, Marcel Dekker, N.Y., N.Y., 1980 (ISBN 0 8247 6918 X), the disclosure of which is incorporated herein by reference in its entirety.

[0283] If the conjugate compound are administered orally, the conjugate compound may be administered, for example, in the form of an aqueous solution or suspension. When aqueous suspensions are prepared for oral use, the active ingredient can be combined with emulsifying and suspending agents. If desired, certain sweetening and / or flavoring agents may be added. The conjugate compound may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be incorporated directly into food. For oral therapeutic administration, the conjugate compound may be incorporated with excipient(s) and used in the form of ingestible elixirs, suspensions, syrups, and the like.

[0284] If the conjugate compound are administered parenterally, the parenteral administration may be by continuous infusion, bolus, or intermittent bolus and may be over a selected period of time. Suitable examples of devices for parenteral administration include needle (including micro needle) injectors, needle free injectors and infusion techniques. For intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared, and the pH of the solutions should be suitably adjusted and buffered. For intravenous use, the total concentration of solutes may be controlled in order to render the preparation isotonic. Thus, in embodiments, one or more of the conjugate compound disclosed herein may be prepared in isotonic medium and administered intravenously.

[0285] The pharmaceutical forms suitable for injectable use may include sterile aqueous solutions or dispersion and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In embodiments, the form is sterile and the fluid is easily syringeable. The preparation of parenteral kits for reconstitution at point-of-care under sterile conditions, for example, by lyophilization, may readily be accomplished using standard pharmaceutical techniques known to those skilled in the art.

[0286] Conjugate compound, including a pharmaceutical composition thereof, for nasal administration may conveniently be formulated as aerosols, drops, gels and powders. Aerosol formulations typically comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which can take the form of a cartridge or refill for use with an atomizing device (e.g. nebuliser, for example, to create a mist-like dispersion of conjugate compound such as an aqueous vehicle (e.g. saline)). Alternatively, the sealed container may be a unitary dispensing device such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use. Where the dosage form comprises an aerosol dispenser, it will contain a propellant which can be a compressed gas such as compressed air or an organic propellant such as fluorochlorohydrocarbon. The aerosol dosage forms can also take the form of a pump-atomizer.

[0287] Conjugate compound, including a pharmaceutical composition thereof, suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, wherein the active ingredient is formulated with a carrier such as sugar, acacia, tragacanth, or gelatin and glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter. In embodiments, a delivery system can be used to deliver the conjugate compound (e.g. formulations or pharmaceutical compositions). It is understood that the delivery system itself may include a device such as an implantable device.

[0288] The conjugate compound may be combined with soluble macromolecular entities, such as cyclodextrin and suitable derivatives thereof or polyethylene glycol containing polymers, in order to improve their solubility, dissolution rate, taste masking, bioavailability and / or stability for use in any of the aforementioned modes of administration. Regardless of the route of administration selected, the conjugate compound which may be used in a suitable hydrated form, and / or the pharmaceutical compositions thereof, are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those of skill in the art.

[0289] In embodiments, the above noted pharmaceutical compositions may be in the form of a controlled release composition, sustained release composition, extended release composition, modified release composition, pulsed release composition, delayed release composition, targeted release composition, site-specific release composition, time release composition, or a combination thereof. To this regard, the term “controlled release” may be variously characterized by "sustained release", “sustained action”, “extended release”, “modified release”, "pulsed release", "delayed release", “targeted release”, “site-specific release”, and “timed release”, which may be used interchangeably herein refer to the time of release, the extent of release, the rate of release, the site of release and / or release of an active ingredient from a composition at such a rate that when a dose of the active ingredient is administered in the sustained release, extended release, pulsed release, timed release, delayed release or controlled-release composition, concentrations (levels) of the active ingredient are maintained within a desired range but below toxic levels over a selected period of time. In the case of in vivo administration, concentrations (levels) of the active ingredient could be measured in blood or plasma, for example. When administered in vivo the sustained release, extended release, pulsed release, timed release, delayed release or controlled-release composition allows for a timely onset of action and useful plasma concentration of an active ingredient to be maintained for longer than in the case of immediate-release forms. The skilled person would understand how the above described formulations may be made into the controlled release composition, sustained release composition, extended release composition, modified release composition, pulsed release composition, delayed release composition, targeted release composition, site-specific release composition, or time release composition.

[0290] Actual dosage levels of the conjugate compound may be varied so as to obtain an amount which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration. To this regard, the dosage of the conjugate compound can depend upon the pharmacokinetic and pharmacodynamic properties of the conjugate compound and its mode and route of administration; the rate of release of the conjugate compound, the age, sex, health, medical condition, the nature and extent of the symptoms and weight of the recipient, the renal and hepatic function of the patient; the frequency of the treatment and the type of concurrent treatment, if any, and the clearance rate of the conjugate compound in the subject to be treated and the effect desired. The selected dosage level may also depend on the additional factors including the activity of the particular conjugate compound and pharmaceutical compositions disclosed herein, the time of administration, the rate of excretion or metabolism of the particular conjugate compound being employed, the rate and extent of absorption, the duration of the treatment, other drugs that may be administered to the patient, compounds and / or materials used in combination with the particular conjugate compound employed and like factors well known in the medical arts. One of skill in the art can determine the appropriate dosage based on the above factors.

[0291] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the conjugate compound or pharmaceutical composition thereof. For example, the physician or veterinarian could start doses of the conjugate compound employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. In general, a suitable daily dose of the conjugate compound will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0292] The conjugate compound may be administered initially in a suitable dosage that may be adjusted as required, depending on the clinical response. In some embodiments, the conjugate compound may be administered in an amount from about 0.001 mg / kg of body weight to about 1000 mg / kg of body weight per day; such as from about 0.01 mg / kg of body weight to about 500 mg / kg of body weight per day; from about 0.01 mg / kg of body weight to about 250 mg / kg of body weight per day; or 0.01 mg / kg of body weight to about 100 mg / kg of body weight per day, and any intermediate ranges or specific amounts, such as from about 0.001 mg / kg, about 0.01 mg / kg, about 0.1 mg / kg, about 1 mg / kg, about 10 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 150 mg / kg, about 200 mg / kg, about 250 mg / kg, about 300 mg / kg, about 350 mg / kg, about 400 mg / kg, about 450 mg / kg, about 500 mg / kg, about 600 mg / kg, about 700 mg / kg, about 800 mg / kg, about 900 mg / kg, or about 1000 mg / kg, to about 0.001 mg / kg, about 0.01 mg / kg, about 0.1 mg / kg, about 1 mg / kg, about 10 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 150 mg / kg, about 200 mg / kg, about 250 mg / kg, about 300 mg / kg, about 350 mg / kg, about 400 mg / kg, about 450 mg / kg, about 500 mg / kg, about 600 mg / kg, about 700 mg / kg, about 800 mg / kg, about 900 mg / kg, or about 1000 mg / kg of body weight per day, per hour, per week, or per dose. If intravenous administration is desired, in typical embodiments, the doses will range from about 0.01 to about 10 mg / kg / minute during a constant rate infusion.

[0293] If formulated as a fixed dose, such combination products employ the conjugate compound within the dosage range described above and the other pharmaceutically active agent(s) within its approved dosage range. The conjugate compound may alternatively be used sequentially with known pharmaceutically acceptable agent(s) when a combination formulation is inappropriate.

[0294] Therapeutically effective amounts of the conjugate compound will generally range up to the maximally tolerated dosage, but may vary widely. The precise amounts employed by the attending physician will vary, of course, depending on the conjugate compound, route of administration, physical condition of the patient (e.g. age, weight, and response of the individual patient, as well as the severity of the patient's symptoms) and other factors. The daily dosage may be administered as a single dosage or may be divided into multiple doses, such as two, three, or four times daily, for administration. Alternatively, the doses may be provided on a weekly, biweekly, or monthly basis. These doses can represent a dosing schedule for a combination therapy of phototherapy and immunotherapy to treat the cancer or the tumour as disclosed herein using the conjugate compound. In some embodiments, reduced dosages may be used as compared to conventional therapeutic dosages of known agents.

[0295] The methods and uses of the conjugate compound and / or composition(s) disclosed herein can allow for enhanced targeting of the conjugate compound and / or composition(s), such that there is improved specificity for the cancer cell(s) and / or tumour(s) through, for example, the TTA disclosed herein. This can allow for precise control over tumour heating and drug release through external thermal excitation. In addition, conjugation of the nanoparticle(s) with the therapeutic agent(s) disclosed herein via the labile linker, can help to minimize off-target effects since the therapeutic agent is guided to the tumour instead of, for example, being systemically released to subsequently destroy non-cancerous cells. In this way, the methods and uses of the conjugate compound and / or composition(s) disclosed herein may contribute to reduced side effects through the targeted and controlled therapeutic agent (e.g. drug molecule) release which can minimize system exposure of the therapeutic agent to healthy cells.

[0296] In other embodiments, there are as follows:

[0297] Embodiment 1 . A conjugate compound comprising: TTA-X1-MNP-X2-TpA wherein: TTA is a tumour targeting agent; MNP is a metal nanoparticle; TpA is a therapeutic agent; and X1and X2are the same or different, and are each independently selected from a non-covalent linker or a covalent linker.

[0298] Embodiment 2. The conjugate compound of embodiment 1 , wherein at least one of X1and X2is a cleavable linker.

[0299] Embodiment 3. The conjugate compound of embodiment 1 or 2, wherein X1or X2is a cleavable linker.

[0300] Embodiment 4. The conjugate compound of any one of embodiments 1 to 3, wherein the cleavable linker is one or more of a chemical cleavable linker, enzyme cleavable linker, heat cleavable linker, pH cleavable linker, and a photochemical cleavable linker.

[0301] Embodiment 5. The conjugate compound of any one of embodiments 1 to 4, wherein the cleavable linker is a photochemical cleavable linker.

[0302] Embodiment 6. The conjugate compound of any one of embodiments 1 to 5, wherein the cleavable linker is cleavable in response to a stimulus.

[0303] Embodiment 7. The conjugate compound of any one of embodiments 1 to 6, wherein the stimulus comprises one or more of photo-irradiation, enzyme(s), nucleophilic / basic reagent(s), reducing agent(s), electrophilic / acidic reagent(s), organometallic and metal reagent(s), and oxidizing reagent(s), heat and pH change.

[0304] Embodiment 8. The conjugate compound of embodiment 7, wherein the stimulus is photo-irradiation.

[0305] Embodiment 9. The conjugate compound of any one of embodiments 1 to 8, wherein the cleavable linker is cleavable at a wavelength in UV, visible, and / or NIR region.

[0306] Embodiment 10. The conjugate compound of any one of embodiments 1 to 9, wherein the cleavable linker is cleavable at a wavelength of about 650 nm to about 1100 nm; about 700 nm to about 1100 nm; about 750 nm to about 1100 nm; about 800 nm to about 1100 nm; about 850 nm to about 1100 nm; about 700 nm to about 1000 nm; about 750 nm to about 950 nm; about 800 nm to about 1000 nm; or about 900 nm to about 1000 nm.

[0307] Embodiment 11 . The conjugate compound of any one of embodiments 1 to 10, wherein X1and X2are each independently selected from at least one base pair of nucleotides; at least two base pairs of nucleotides; at most 50 base pairs of nucleotides; at most 40 base pairs of nucleotides; at most 30 base pairs of nucleotides; at most 20 base pairs of nucleotides; or about 2 to about 20 base pairs of nucleotides.

[0308] Embodiment 12. The conjugate compound of any one of embodiments 1 to 11 , wherein X1and X2are each independently selected from a double strand of complementary polynucleotide or double strand of complementary oligonucleotide or a double strand of complementary DNA strands.

[0309] Embodiment 13. The conjugate compound of any one of embodiments 1 to 12, wherein X1and X2are each independently selected from DNA and / or RNA with a suitable sequence and length.

[0310] Embodiment 14. The conjugate compound of any one of embodiments 1 to 13, wherein X1and X2are each independently selected from a linker with a linking functional group (e.g. amide, esters, ethers, thioethers, thioester, dative bond, carbamate, Schiff base, secondary amine, hydrazone, oxime, diazo bond, isourea, isothiourea, sulfonamide, aryl amine, amidine, phosphoramidate, hydroxyl, alkene, alkyne, thiol, disulfide, etc.).

[0311] Embodiment 15. The conjugate compound of any one of embodiments 1 to 14, wherein X1and X2are each independently selected from valine-citruline, valine-alanine, or a combination of two to eight amino acids; a self-immolative unit (e.g., a PAB spacer, PEG spacer, etc.), and optionally hydrophilic groups (e.g., PEG); glucuronides, peptides, or a linker with a terminal maleimide group, hydrazone, and / or disulfide; and / or a non-covalent interaction such as hydrogen bonds.

[0312] Embodiment 16. The conjugate compound of any one of embodiments 1 to 15, wherein X1and X2are each independently selected from S or FcS.

[0313] Embodiment 17. The conjugate compound of any one of embodiments 1 to 16, wherein the MNP is a tube, rod, shell, cage, sphere, fiber, wire, stars, plate, sea archon, or a combination thereof.

[0314] Embodiment 18. The conjugate compound of any one of embodiments 1 to 17, wherein the MNP has an average particle size that is less than about 1 pm in size, and typically from about 1 nm to about 900 nm in size and various ranges therebetween. Embodiment 19. The conjugate compound of any one of embodiments 1 to 18, wherein the average particle size of the MNP is about 1 nm to about 100 nm.

[0315] Embodiment 20. The conjugate compound of any one of embodiments 1 to 19, wherein the MNP has one or more dimensions of the order of 100 nm or less.

[0316] Embodiment 21 . The conjugate compound of any one of embodiments 1 to 20, wherein the MNP is a metal nanorod (MNR).

[0317] Embodiment 22. The conjugate compound of embodiment 21 , wherein the MNR ranges from about 1 to about 100 nm.

[0318] Embodiment 23. The conjugate compound of embodiment 21 or 22, wherein the MNR has a diameter or cross-section of between about 5 nm and about 50 nm, such as from about 5 nm, about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, or about 45 nm, to about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, about 45 nm, or about 50 nm.

[0319] Embodiment 24. The conjugate compound of any one of embodiments 21 to 23, wherein the MNR has an axial length of between about 20 nm and about 500 nm, such as from about 20 nm, about 30 nm, about 40 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 125 nm, about 150 nm, about 175 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, or about 450 nm, to about 30 nm, about 40 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 125 nm, about 150 nm, about 175 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, or about 500 nm.

[0320] Embodiment 25. The conjugate compound of any one of embodiments 21 to 24, wherein the MNR has an aspect ratio of from about 1.1 to about 100, about 1.1 , about 1.2, about 1.3, about 1 .4, about 1 .5, about 1 .6, about 1 .7, about 1 .8, about 1.9, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, or about 90, to about 1.2, about 1 .3, about 1 .4, about 1.5, about 1 .6, about 1 .7, about 1 .8, about 1 .9, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, or from about 1.1 to about 10.

[0321] Embodiment 26. The conjugate compound of any one of embodiments 21 to 25, wherein the MNR have a variance for the length, diameter, and / or aspect ratio of any given metal nanorod in a population or subpopulation can be at most 10%, 8%, 5%, 2%, 1% or 0.1% different from the average length, diameter, and / or aspect ratio for metal nanorods in the population.

[0322] Embodiment 27. The conjugate compound of any one of embodiments 21 to 26, wherein the MNR the population can be composed of at least 90%, 95%, 99% or 99.9% metal nanorods having a particular length, diameter, and / or aspect ratio.

[0323] Embodiment 28. The conjugate compound of any one of embodiments 21 to 27, wherein the metal is a transition metal, a precious metal, or a combination thereof.

[0324] Embodiment 29. The conjugate compound of any one of embodiments 1 to 28, wherein the metal is selected from gold, nickel, palladium, platinum, copper, silver, zinc, cadmium, or a combination thereof.

[0325] Embodiment 30. The conjugate compound of any one of embodiments 1 to 29, wherein the metal is gold.

[0326] Embodiment 31 . The conjugate compound of any one of embodiments 1 to 30, wherein the MNP is a gold nanorod (GNR).

[0327] Embodiment 32. The conjugate compound of any one of embodiments 1 to 31 , wherein the MNP is capped with any suitable capping agents such as, and without being limited thereto, carboxylic acid, conventional citrate, and / or a positively charged ligand.

[0328] Embodiment 33. The conjugate compound of any one of embodiments 1 to 32, wherein the MNP is wrapped comprising a polymer (e.g. PEG).

[0329] Embodiment 34. The conjugate compound of any one of embodiments 1 to 33, wherein the TTA is an agent for recognizing a target cell.

[0330] Embodiment 35. The conjugate compound of any one of embodiments 1 to 34, wherein the compound accumulates in cancer (e.g. tumour) with the assistance from an EPR (enhanced permeability effect).

[0331] Embodiment 36. The conjugate compound of any one of embodiments 1 to 35, wherein the compound accumulates excessively in cancer (e.g. tumour) compared to healthy tissue.

[0332] Embodiment 37. The conjugate compound of any one of embodiments 1 to 36, wherein the TTA reduces toxicity from off-target delivery of the TpA.

[0333] Embodiment 38. The conjugate compound of any one of embodiments 1 to 37, wherein the target cell is a cancer cell.

[0334] Embodiment 39. The conjugate compound of any one of embodiments 1 to 38, wherein the target cell is a tumour cell. Embodiment 40. The conjugate compound of any one of embodiments 1 to 39, wherein the TTA comprises a peptide, a protein, a polypeptide, an antibody or functional equivalent thereof, an antibody fragment, nucleic acids, nucleosides, aptamers, cell receptor inhibitors, hormones, or a combination thereof.

[0335] Embodiment 41 . The conjugate compound of any one of embodiments 1 to 40, wherein the TTA comprises an antibody, or a functional equivalent thereof.

[0336] Embodiment 42. The conjugate compound of embodiment 41 , wherein the antibody or functional equivalent thereof is anti-CD20, anti-PSMA, PSMA binding motif (Glu-urea-Lys), anti-CD47, anti-EGFR or a combination thereof.

[0337] Embodiment 43. The conjugate of any one of embodiments 1 to 41 , wherein the TTA targets 1GH-IGK, 43-9F, 5T4, 791Tgp72, acyclophilin C-associated protein, alphafetoprotein (AFP), a-actinin-4, A3, antigen specific for A33 antibody, ART-4, B7, Ba 733, BAGE, BCMA, BCR-ABL, beta-catenin, beta-HCG, BrE3-antigen, BCA225, BTAA, CA125, CA 15-3\CA 27.29\BCAA, CA195, CA242, CA-50, CAM43, CAMEL, CAP-1 , carbonic anhydrase IX, c-Met, CA19-9, CA72-4, CAM 17.1 , CASP-8 / m, CCCL19, CCCL21 , CD1 , CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, CD20, CD21 , CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD44, CD45, CD46, CD47, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD68, CD70, CD70L, CD74, CD79a, CD79b, CD80, CD83, CD95, CD126, CD132, CD133, CD138, CD147, CD154, CDC27, CDK4, CDK4m, CDKN2A, CO-029, CTLA4, CXCR4, CXCR7, CXCL12, HIF-1a, colon-specific antigen-p (CSAp), CEA (CEACAM5), CEACAM6, c-Met, DAM, E2A-PRL, EGFR, EGFRvlll, EGP-1 (TROP-2), EGP-2, ELF2-M, Ep-CAM, fibroblast growth factor (FGF), FGF-5, Flt-1 , Flt-3, folate receptor, G250 antigen, Ga733VEpCAM, GAGE, gp100, GRO- , H4-RET, HLA-DR, HM1.24, human chorionic gonadotropin (HCG) and its subunits, HER2 / neu, HMGB-1 , hypoxia inducible factor (HIF- 1), HSP70-2M, HST-2, HTgp-175, la, IGF-1 R, IFN-y, IFN-a, IFN- , IFN-A, IL-4R, IL-6R, IL- 13R, IL-15R, IL-17R, IL-18R, IL-2, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-23, IL-25, insulinlike growth factor-1 (IGF-1), KC4-antigen, KSA, KS-1-antigen, KS1-4, LAGE-1a, Le-Y, LDR / FUT, M344, MA-50, macrophage migration inhibitory factor (MIF), MAGE, MAGE-1 , MAGE-3, MAGE-4, MAGE-5, MAGE-6, MART- 1 , MART-2, TRAG-3, mCRP, MCP-1 , MIP- 1A, MIP-1 B, MIF, MG7-Ag, MOV18, MUC1 , MUC2, MUC3, MUC4, MUC5ac, MUC13, MUC16, MUM-1 / 2, MUM-3, MYL-RAR, NB / 70K, Nm23H1 , NuMA, NCA66, NCA95, NCA90, NY-ESO-1 , p15, p16, p185erbB2, p180erbB3, PAM4 antigen, pancreatic cancer mucin, PD1 receptor (PD-1), PD-1 receptor ligand 1 (PD-L1), PD-1 receptor ligand 2 (PD-L2), PI5, placental growth factor, p53, PLAGL2, Pmel17 prostatic acid phosphatase, PSA, PRAME, PSMA, P1GF, ILGF, ILGF-1 R, IL-6, IL-25, RCAS1 , RS5, RAGE, RANTES, Ras, T101 , SAGE, S100, survivin, survivin-2B, SDDCAGi6, TA-90Mac2 binding protein, TAAL6, TAC, TAG-72, TLP, tenascin, TRAIL receptors, TRP-1 , TRP-2, TSP-180, TNF-a, Tn antigen, Thomson-Friedenreich antigens, tumor necrosis antigens, tyrosinase, VEGFR, ED-B fibronectin, WT-1 , 17-1A-antigen, complement factors C3, C3a, C3b, C5a, C5, an angiogenesis marker, bcl-2, bcl-6, K-ras, or any combination thereof.

[0338] Embodiment 44. The conjugate compound of any one of embodiments 1 to 43, wherein the TpA comprises an agent that treats cancer.

[0339] Embodiment 45. The conjugate compound of any one of embodiments 1 to 44, wherein the TpA comprises a protein, peptide, nucleic acid, amino acid, nucleoside, antibody, antibody drug conjugate (ADC), antibody fragment, antibody ligand, peptide nucleic acid, small organic molecule, lipid, hormone, drug, enzyme, lectin, cell adhesion molecule, antibody epitope, enzyme substrate, enzyme inhibitor, coenzyme, organic molecule, carbohydrate, such as polysaccharides and monosaccharides, or a combination thereof.

[0340] Embodiment 46. The conjugate compound of any one of embodiments 1 to 45, wherein the TpA comprises a toll like receptor (TLR) agonist, immunomodulator, drug molecule or a combination thereof.

[0341] Embodiment 47. The conjugate compound of embodiment 46, wherein the immunomodulator comprises a cytokine, cytokine agonists, a chemokine, chemokine agonists, a toll-like receptor (TLR) agonists, or a combination thereof.

[0342] Embodiment 48. The conjugate compound of embodiment 47, wherein the TLR agonist comprises a TLR-4 agonist, TLR-7 agonist, TLR-8 agonist, TLR-9 agonist, or a combination thereof.

[0343] Embodiment 49. The conjugate compound of any one of embodiments 46 to 48, wherein the immunomodulator comprises a cytokine.

[0344] Embodiment 50. The conjugate compound of embodiment 49, wherein the cytokine comprises IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12 IL-15, IL-17, IL-23, IL-10, TNF-a, IFN-a, IFN-0, IFN-y, or a combination thereof.

[0345] Embodiment 51 . The conjugate compound of any one of embodiments 46 to 50, wherein the immunomodulator comprises a chemokine, such as CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CXCL11 , CXCL12, CXCL14, CCL2, CCL5, or a combination thereof.

[0346] Embodiment 52. The conjugate of any one of embodiments 46 to 52, wherein the immunomodulator comprises a checkpoint inhibitor. Embodiment 53. The conjugate compound of embodiment 52, wherein the checkpoint inhibitor comprises a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, or a combination thereof.

[0347] Embodiment 54. The conjugate compound of embodiment 53, wherein the checkpoint inhibitor comprises nivolumab, pembrolizumab, cemiplimab, ipilimumab, atezolizumab, avelumab, durvalumab, relatlimab, or a combination thereof.

[0348] Embodiment 55. The conjugate compound of any one of embodiments 1 to 54, wherein the TpA comprises a chemotherapeutic agent.

[0349] Embodiment 56. The conjugate compound of any one of embodiments 1 to 55, wherein TpA comprises Temozolomide, Actinomycin, Alitretinoin, All-trans retinoic acid, Azacitidine, Azathioprine, Bevacizumab, Bexatotene, Bleomycin, Bortezomib, Carboplatin, Capecitabine, Cetuximab, Cisplatin, Chlorambucil, Cyclophosphamide, Cytarabine, Daunorubicin, Docetaxel, Doxifluridine, Doxorubicin, Epirubicin, Epothilone, Erlotinib, Etoposide, Fluorouracil, Gefitinib, Gemcitabine, Hydroxyurea, Idarubicin, Imatinib, Ipilimumab, Irinotecan, Mechlorethamine, Melphalan, Mercaptopurine, Methotrexate, Mitoxantrone, Ocrelizumab, Ofatumumab, Oxaliplatin, Paclitaxel, Panitumab, Pemetrexed, Rituximab, Tafluposide, Teniposide, Tioguanine, Topotecan, Tretinoin, Valrubicin, Vemurafenib, Vinblastine, Vincristine, Vindesine, Vinorelbine, Vorinostat, Romidepsin, 5- fluorouracil (5-FU), 6-mercaptopurine (6-MP), Cladribine, Clofarabine, Floxuridine, Fludarabine, Pentostatin, Mitomycin, ixabepilone, Estramustine, prednisone, methylprednisolone, dexamethasone, or a combination thereof.

[0350] Embodiment 57. The conjugate compound of any one of embodiments 1 to 56, wherein the conjugate compound is wrapped with a polymer (e.g. PEG).

[0351] Embodiment 58. The conjugate compound of any one of embodiments 1 to 57, wherein the conjugate compound comprises PSMABM-X1-GNR-X2-IL2, CD47-X1-GNR-X2- IL2, PSMABM-X1-GNR-X2-PD1 , CD47-X1-GNR-X2-PD1 , or a combination thereof.

[0352] Embodiment 59. The conjugate compound of embodiment 58, wherein X1and X2are independently selected from S, ssDNA, dsDNA, or S-Fc.

[0353] Embodiment 60. The conjugate compound of embodiment 58 or 59, wherein the conjugate compound comprises PSMABM-GNR-IL2, CD47-GNR-IL2, PSMABM-GNR- PD1 , CD47-GNR-PD1 , PSMABM-S-GNR-SFclL2, CD47-S-GNR-SFclL2, PSMABM-S- GNR-SFcPD1 , CD47-S-GNR-SFcPD1 , or a combination thereof. Embodiment 61 . The conjugate compound of any one of embodiments 1 to 60, wherein the conjugate compound comprises TTA-FcS-GNR-S-TpA, TTA-GNR-S-TpA, TTA-FcS-GNR-TpA, or a combination thereof.

[0354] Embodiment 62. The conjugate compound of any one of embodiments 1 to 61 , wherein the conjugate compound is capable of moving intratumorally.

[0355] Embodiment 63. The conjugate compound of any one of embodiments 1 to 62 for treatment of cancer.

[0356] Embodiment 64. The conjugate compound of embodiment 63, wherein the cancer is a tumour.

[0357] Embodiment 65. The conjugate compound of embodiment 64, wherein the cancer is a solid tumour.

[0358] Embodiment 66. The conjugate compound of any one of embodiments 1 to 65, for use in combination therapy, optionally, phototherapy, photothermaltherapy and immunotherapy.

[0359] Embodiment 67. A pharmaceutical composition comprising the conjugate compound of any one of embodiments 1 to 66.

[0360] Embodiment 68. The pharmaceutical composition of embodiment 67, wherein the composition is a dispersion, optionally, a colloidal dispersion.

[0361] Embodiment 69. The pharmaceutical composition of embodiment 67 or 68, further comprising pharmaceutically acceptable excipient(s).

[0362] Embodiment 70. The pharmaceutical composition according to any one of embodiments 67 to 69 for treatment of cancer.

[0363] Embodiment 71 . The pharmaceutical composition of embodiment 70, wherein the cancer is a tumour.

[0364] Embodiment 72. The pharmaceutical composition of embodiment 71 , wherein the cancer is a solid tumour.

[0365] Embodiment 73. The pharmaceutical composition of any one of embodiments 67 to 72 for use in combination therapy, optionally, phototherapy, photothermaltherapy and immunotherapy.

[0366] Embodiment 74. A method for making the compound of any one of embodiments 1 to 66, the method comprising: combining TTA-CRG1 and CRG2-MNP-X2-TpA to make TTA- X1-MNP-X2-TpA, wherein CRG is a conjugate reactive group. Embodiment 75. The method of embodiment 74, the method further comprises combining MNP-X2-TpA and CRG2 to make the CRG2-MNP-X2-TpA.

[0367] Embodiment 76. A method for making the compound of any one of embodiments 1 to 66, the method comprising: combining TTA-X1-MNP-CRG3 and CRG4-TpA to make TTA- X1-MNP-X2-TpA, wherein CRG3 and CRG4 are different, and are each selected to form X2, optionally, CRG4 is a portion of the TpA, wherein CRG is a conjugate reactive group.

[0368] Embodiment 77. The method of embodiment 76, wherein the method further comprises combining TTA-X1-MNP and CRG3 to make the TTA-X1-MNP-CRG3.

[0369] Embodiment 78. A method for making the compound of any one of embodiments 1 to 66, the method comprising: combining TTA-CRG1 , CRG2-MNP-CRG3, and CRG4-TpA to make TTA-X1-MNP-X2-TpA, wherein CRG is a conjugate reactive group.

[0370] Embodiment 79. The method of embodiment 78, wherein the method further comprises combining TTA-CRG1 , CRG2-MNP-CRG3, and CRG4-TpA, in any order.

[0371] Embodiment 80. The method of embodiment 78 or 79, wherein the method further comprises combining TTA-X1-MNP and CRG3 to make TTA-X1-MNP-CRG1 .

[0372] Embodiment 81 . The method of any one of embodiments 74 to 80, wherein CRG1 and CRG2 are different, and are each selected from at least one nucleotide, forming at least one base pair of nucleotides.

[0373] Embodiment 82. The method of any one of embodiments 74 to 81 , wherein CRG1 and CRG2 are different, and are each selected from at least two nucleotides, forming at least two base pairs of nucleotides.

[0374] Embodiment 83. The method of any one of embodiments 74 to 82, wherein CRG3 and CRG4 are different, and are each selected from at least one nucleotide, forming at least one base pair of nucleotides.

[0375] Embodiment 84. The method of any one of embodiments 74 to 83, wherein CRG3 and CRG4 are different, and are each selected from at least two nucleotides, forming at least two base pairs of nucleotides.

[0376] Embodiment 85. The method of any one of embodiments 74 to 84, wherein the at least two nucleotides are polynucleotides (e.g. nucleic acids) or oligonucleotides.

[0377] Embodiment 86. The method of any one of embodiments 74 to 85, wherein CRG1 and CRG2 are each complement ssDNA.

[0378] Embodiment 87. The method of any one of embodiments 74 to 86, wherein CRG3 and CRG4 are each complement ssDNA. Embodiment 88. The method of any one of embodiments 74 to 87, wherein CRG1 and CRG2 are each complement ssDNA that only bind to one another.

[0379] Embodiment 89. The method of any one of embodiments 74 to 88, wherein CRG3 and CRG4 are each complement ssDNA that only bind to one another.

[0380] Embodiment 90. The method of any one of embodiments 74 to 89, wherein CRG1 and CRG2 are different, and are each selected from a moiety or group capable of forming a covalent linker as a result of the association between atoms or molecules of each CRG 1 and CRG2, forming a linking functional group.

[0381] Embodiment 91 . The method of any one of embodiments 74 to 90, wherein CRG3 and CRG4 are different, and are each selected from a moiety or group capable of forming a covalent linker as a result of the association between atoms or molecules of each CRG3 and CRG4, forming a linking functional group.

[0382] Embodiment 92. The method of any one of embodiments 74 to 91 , wherein CRG1 to CRG4 are different, and are each selected from (a) carboxyl groups and various derivatives thereof including, but not limited to, N-hydroxysuccinimide esters (e.g. sulfo-N- hydroxysuccinimide), N-hydroxybenztriazole esters, acid halides, acyl imidazoles, thioesters, p-nitrophenyl esters, alkyl, alkenyl, alkynyl and aromatic esters; (b) hydroxyl groups which can be converted to esters, ethers, aldehydes, etc.; (c) haloalkyl groups wherein the halide can be later displaced with a nucleophilic group such as, for example, an amine, a carboxylate anion, thiol anion, carbanion, or an alkoxide ion, thereby resulting in the covalent attachment of a new group at the site of the halogen atom; (d) dienophile groups which are capable of participating in Diels-Alder reactions such as, for example, maleimido or maleimide groups; (e) aldehyde or ketone groups such that subsequent derivatization is possible via formation of carbonyl derivatives such as, for example, imines, hydrazones, semicarbazones or oximes, or via such mechanisms as Grignard addition or alkyllithium addition; (f) sulfonyl halide groups for subsequent reaction with amines, for example, to form sulfonamides; (g) thiol groups, which can be converted to disulfides, reacted with acyl halides, or bonded to metals such as gold, or react with maleimides; (h) amine or sulfhydryl groups (e.g., present in cysteine), which can be, for example, acylated, alkylated or oxidized; (i) alkenes, which can undergo, for example, cycloadditions, acylation, Michael addition, etc.; (j) epoxides, which can react with, for example, amines and hydroxyl compounds; (k) phosphoramidites and other standard functional groups useful in nucleic acid synthesis; (I) metal silicon oxide bonding; (m) metal bonding to reactive phosphorus groups (e.g., phosphines) to form, for example, phosphate diester bonds; (n) azides coupled to alkynes using copper catalyzed cycloaddition click chemistry; (o) biotin conjugate can react with avidin or strepavidin to form a avidin-biotin complex or streptavidin-biotin complex.

[0383] Embodiment 93. The method of any one of embodiments 74 to 92, wherein at least one of the CRGs are zero length linkers.

[0384] Embodiment 94. The method of any one of embodiments 74 to 93, further comprising adding a cross-linker between at least two CRGs.

[0385] Embodiment 95. The method of any one of embodiments 74 to 94, wherein TTA and / or TpA have at least one nucleotide to complement with CRG2 and CRG3, respectively.

[0386] Embodiment 96. A method for treating cancer, comprising administering to a mammal a therapeutically effective amount of the conjugate compound according to any one of embodiments 1 to 66 or the composition according to any one of embodiments 67 to 73.

[0387] Embodiment 97. The method of embodiment 96, wherein the conjugate compound or the composition targets the cancer (e.g. cancer cells).

[0388] Embodiment 98. The method of embodiment 96 or 97, wherein the TTA binds to the cancer cell.

[0389] Embodiment 99. The method of any one of embodiments 96 to 98, wherein the TTA binds to a tumour.

[0390] Embodiment 100. The method of any one of embodiments 96 to 99, wherein the TTA binds to a solid tumour.

[0391] Embodiment 101. The method of any one of embodiments 96 to 100, further comprising cleaving X1or X2.

[0392] Embodiment 102. The method of any one of embodiments embodiment 101 , wherein the cleaving comprises one or more of a chemical cleaving, enzyme cleaving, heat cleaving, pH cleaving, and a photochemical cleaving.

[0393] Embodiment 103. The method of embodiment 101 or 102, wherein cleaving comprises applying a stimulus to the conjugate compound or the composition.

[0394] Embodiment 104. The method of embodiment 103, wherein the stimulus comprises pH, enzymes, electromagnetic radiation (EMR), or a combination thereof.

[0395] Embodiment 105. The method of embodiment 104, wherein the stimulus comprises EMR. Embodiment 106. The method of embodiment 104 or 105, wherein the EMR is selected from near-infrared, visible light, and UV, or a combination thereof.

[0396] Embodiment 107. The method of embodiment 106, wherein the EMR is near-infrared.

[0397] Embodiment 108. The method of any one of embodiments 104 to 107, wherein the EMR emits a wavelength in a range of about 700 nm to about 1000 nm, about 800 nm to about 1000 nm, about 900 nm to about 1000 nm, or about 950 nm to about 1000 nm.

[0398] Embodiment 109. The method of any one of embodiments 104 to 108, wherein the EMR excites the conjugate compound or the composition to generate heat.

[0399] Embodiment 110. The method of any one of embodiments 104 to 109, wherein the EMR cleaves X2to release the TpA.

[0400] Embodiment 111. The method of any one of embodiments 96 to 110, wherein the conjugate compound or composition generates heat to reduce growth of the tumour.

[0401] Embodiment 112. The method of any one of embodiments 104 to 111 , wherein the EMR is generated by a light-emitting diode (LED) or a laser generator.

[0402] Embodiment 113. The method of any one of embodiments 96 to 112, wherein the cancer is selected from brain tumour, nerve sheath tumour, breast cancer, colon cancer, colorectal cancer, skin cancer, lung cancer, hepatocellular cancer, gastric cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, cancer of the urinary tract, thyroid cancer, renal cancer, renal cell carcinoma, carcinoma, melanoma, lymphoma, head and neck cancer, brain cancer, and / or prostate cancer, including but not limited to androgen-dependent prostate cancer and androgen-independent prostate cancer.

[0403] Embodiment 114. The method of any one of embodiments 96 to 113, wherein the cancer is selected from prostate and / or breast cancer.

[0404] Embodiment 115. The method of any one of embodiments 96 to 114, wherein the cancer is a tumour.

[0405] Embodiment 116. The method of any one of embodiments 96 to 115, wherein the cancer is a solid tumour.

[0406] Embodiment 117. The method of any one of embodiments 96 to 116, administering the conjugate compound or composition as a combined therapy, optionally, photothermaltherapy and immunotherapy.

[0407] Embodiment 118. The method of any one of embodiments 96 to 117, wherein sideeffects generated from off-target delivery and / or actions of the TpA is reduced. Embodiment 119. The method of any one of embodiments 96 to 118, wherein the conjugate compound or the composition has a synergistic effect.

[0408] Embodiment 120. The method of any one of embodiments 96 to 119, wherein the conjugate compound or the composition has an additive effect.

[0409] Embodiment 121 . The method of any one of embodiments 96 to 120, wherein the mammal is a human.

[0410] Embodiment 122. The method of any one of embodiments 96 to 121 , wherein administering comprises parenteral administration (e.g. formulated for injection via the intravenous, intramuscular, sub-cutaneous, intralesional, and / or intraperitoneal routes).

[0411] Embodiment 123. The method of any one of embodiments 96 to 122, wherein administering comprises a single injection for treatment versus two separate injections of TTA-MNP and TpA-MNP.

[0412] Embodiment 124. Use of a therapeutically effective amount of the conjugate compound according to any one of embodiments 1 to 66 or the composition according to any one of embodiments 67 to 73 for treating cancer.

[0413] Embodiment 125. The use of embodiment 124, wherein the conjugate compound or the composition targets the cancer (e.g. cancer cells).

[0414] Embodiment 126. The use of embodiment 124 or 125, wherein the TTA of the conjugate binds to the cancer cell.

[0415] Embodiment 127. The use of any one of embodiments 124 to 126, wherein the TTA binds to a tumour.

[0416] Embodiment 128. The use of any one of embodiments 124 to 127, wherein the TTA binds to a solid tumour.

[0417] Embodiment 129. The use of any one of embodiments 124 to 128, wherein X1or X2are cleavable.

[0418] Embodiment 130. The use of embodiment 129, wherein X1or X2are one or more of chemical cleavable, enzyme cleavable, heat cleavable, pH cleavable, and photochemical cleavable.

[0419] Embodiment 131. The use of any one of embodiments 124 to 130, wherein X1or X2are cleavable by application of a stimulus to the conjugate compound or the composition.

[0420] Embodiment 132. The use of embodiment 131 , wherein the stimulus comprises pH, enzymes, electromagnetic radiation (EMR), or a combination thereof. Embodiment 133. The use of embodiment 132, wherein the stimulus comprises EMR.

[0421] Embodiment 134. The use of embodiment 132 or 133, wherein the EMR is selected from near-infrared, visible light, and UV, or a combination thereof.

[0422] Embodiment 135. The use of embodiment 134, wherein the EMR is near-infrared.

[0423] Embodiment 136. The use of any one of embodiments 132 to 135, a wavelength in a range of about 700 nm to about 1000 nm, about 800 nm to about 1000 nm, about 900 nm to about 1000 nm, or about 950 nm to about 1000 nm.

[0424] Embodiment 137. The use of any one of embodiments 132 to 136, wherein the EMR excites the conjugate compound or the composition to generate heat.

[0425] Embodiment 138. The use of any one of embodiments 132 to 137, wherein the EMR cleaves X2to release the TpA.

[0426] Embodiment 139. The use of any one of embodiments 124 to 138, wherein the conjugate compound or composition generates heat to reduce growth of the tumour.

[0427] Embodiment 140. The use of any one of embodiments 124 to 139, wherein the EMR is generated by a light-emitting diode (LED) or a laser generator.

[0428] Embodiment 141 . The use of any one of embodiments 124 to 140, wherein the cancer is selected from brain tumour, nerve sheath tumour, breast cancer, colon cancer, colorectal cancer, skin cancer, lung cancer, hepatocellular cancer, gastric cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, cancer of the urinary tract, thyroid cancer, renal cancer, renal cell carcinoma, carcinoma, melanoma, lymphoma, head and neck cancer, brain cancer, and / or prostate cancer, including but not limited to androgen-dependent prostate cancer and androgen-independent prostate cancer.

[0429] Embodiment 142. The use of any one of embodiments 124 to 141 , wherein the cancer is selected from prostate and / or breast cancer.

[0430] Embodiment 143. The use of any one of embodiments 124 to 142, wherein the cancer is a tumour.

[0431] Embodiment 144. The use of any one of embodiments 124 to 143, wherein the cancer is a solid tumour.

[0432] Embodiment 145. The use of any one of embodiments 124 to 144, the conjugate compound or composition is used as a combined therapy, optionally, phototherapy and immunotherapy.

[0433] Embodiment 146. The use of any one of embodiments 124 to 145, wherein side-effects generated from off-target delivery and / or actions of the TpA is reduced. Embodiment 147. The use of any one of embodiments 124 to 146, wherein the conjugate compound or composition has a synergistic effect.

[0434] Embodiment 148. The method of any one of embodiments 124 to 147, wherein the conjugate compound or the composition has an additive effect.

[0435] Embodiment 149. The use of any one of embodiments 124 to 148, wherein the mammal is a human.

[0436] Embodiment 150. The use of any one of embodiments 124 to 149, wherein the conjugate compound or composition is for parenteral administration (e.g. formulated for injection via the intravenous, intramuscular, sub-cutaneous, intralesional, and / or intraperitoneal routes).

[0437] Embodiment 151. The use of any one of embodiments 124 to 141 , wherein the conjugate compound or composition is a single injection for treatment versus two separate injections of TTA-MNP and TpA-MNP.

[0438] The above disclosure generally describes the present invention. A more complete understanding can be obtained by reference to the following specific Examples. The Examples are described solely for purposes of illustration and are not intended to limit the scope of the invention. Changes in form and substitution of equivalents are contemplated as circumstances may suggest or render expedient. Although specific terms have been employed herein, such terms are intended in a descriptive sense and not for purposes of limitation.

[0439] EXAMPLES

[0440] 1) TTA-X1-GNR

[0441] GNRs are selected from GNRs having a wavelength in the absorption range of about 650 nm to about 1100 nm based on longitudinal surface plasmon resonance (LSPR). In certain examples, the GNRs are selected from GNRs having a wavelength in the absorption range of about 800 nm to about 850 nm of LSPR.

[0442] The TTA is an antibody selected from anti-CD20 and anti-PSMA or an antibody fragment or an antibody mimic or a poly-peptide or a protein (somatostatin receptor binding molecules and somatostatin subtypes), or hormones. The TTA can carry a natural or synthetic flanking functional group (e.g. amine, carboxylic acid, or ester) for conjugation with GNRs using any of the methods described below. One skilled in the art would understand how to select the following methods depending on the final use / required treatment. i) Non-Covalent Linker

[0443] Conjugation using a non-covalent linker to form TTA-GNR:

[0444] Passively bind TTA to GNRs by mixing a solution of TTA in a buffer with a solution of GNRs in triply deionised water (TWD) whereby the concentration of the TTA in the final solution is about 0.1 to about 3 mg / mL The optical density (OD) of the GNRs in solution is about 1.0 to about 10.0. Depending upon the selected TTA, the incubation time and temperature is suitable for the specific TTA. In examples, the incubation time is about 0.25 to about 24 hrs and the temperature is about 2°C to about 30°C, respectively. Excess TTA is removed with centrifugation and exchange of supernatant to form TTA-GNR (Figure 2 ). ii) Covalent Linker

[0445] Conjugation using a covalent linker to form TTA-C(O)NH2-GNR:

[0446] Use carbodiimide 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDO) and N- hydroxysuccinimide (NHS) linkage chemistry (or amide linkage) for TTA and GNR conjugation. For this, carboxylic acid functionalization of GNRs can be achieved using literature methods (see Hermanson, G.T., “Bioconjugation Techniques”, 3rd edition) such as wrapping GNRs with a carboxyl ending polymer (e.g. thiolated carboxy polyethylene glycol) or capping GNRs with a molecule with carboxylic acid at its one end and thiol or disulfide group on its other end for binding to GNRs (e.g. lipoic acid). See Figure 3, where 1 is GNR and 2 is TTA.

[0447] A solution of lipoic acid is mixed with GNR solution to provide a concentration of about 0.1 to about 10 mg / mL The optical density (OD) of the GNRs in the solution is about 1 .0 to about 10.0. Depending upon the total volume of the solution, the incubation time can vary from about 1 hr to about 24 hrs at room temperature. Excess unbounded lipoic acid is removed using a centrifuge method, for this, mixed solution is centrifuged first at about 5000 to about 20,000 ref for about 5 to about 15 minutes and then the supernatant is replaced with TDW.

[0448] The lipoic acid treated GNRs is mixed with a solution of TTA in a buffer in triply deionised water (TWD) whereby the concentration of the TTA in the final solution is about 0.1 to about 3 mg / mL The optical density (OD) of the GNRs in solution is about 1 .0 to about 10.0. Depending upon the selected TTA, the incubation time and temperature is suitable for the specific TTA. In examples, the incubation time is about 0.25 to about 24 hrs and the temperature is about 2°C to about 30°C, respectively. Excess TTA is removed with centrifugation and exchange of supernatant to form TTA-C(O)NH2-GNR (Figure 3 ). iii) Non-Covalent Linker (hydrogen bonding)

[0449] Conjugation using two complementary ssDNA to form TTA-DNA(1)-GNR:

[0450] A conjugate of GNRs with a ssDNA (ssDNA-GNRs) is formed. Mixing a dilute solution of thiolated ssDNA in Tris-EDTA buffer with a pH of about 7 to about 8 and having a buffer strength of about 5 mM to about 50 mM, with GNRs solution in TDW to produce their respective concentrations of about 0.01 to about 1.0 ug / mL and about 1.0 to about 10 OD in the final solution (ssDNA-GNR) (Ab-ssDNA). Depending upon the total volume of the solution, the incubation time can vary from about 1 hr to about 24 hrs at a temperature of about 2°C to about 25°C is provided. Excess unbound ssDNA is removed using a centrifuge method. The solution is centrifuged first at about 5000 to about 20,000 ref for about 5 to about 15 minutes and then the supernatant is replaced with equal amounts of Tris-EDTA buffer as used to make ssDNA solution.

[0451] Secondly, TTA is conjugated to complementary ssDNA to form ssDNA-TTA conjugate solution by using a method available in the literature (Hermanson, G.T., “Bioconjugation Techniques”, 3rd edition, page 157-164) or using commercial antibody- oligonucleotide conjugation kits (AbCam™ or AlphaThera™). Excess unbound ssDNA is removed using chromatography such as affinity column chromatography. ssDNA-GNRs conjugate solution is mixed with ssDNA-TTA solution in unequal amounts with about 0.1 to about 5% higher amount of ssDNA-TTA for either enzymatic in the presence of DNA Ligase non-enzymatic nucleic acid winding / zipping / clipping. Depending upon the total volume of the solution, the incubation time can vary from about 1 hr to about 24 hrs at room temperature is provided. Excess unbound ssDNA is removed and active enzyme (if enzymatic zipping / clipping is used) is removed using centrifuge method or tangential flow filtration (TFF) or combination of both to form TTA-DNA(1)-GNR (Figure 4).

[0452] 2) TPA-X2-GNR

[0453] Methods for Conjugation of TpA-ssDNA and conjugation to complementary ssDNA-GNR TpAs are selected from toll like receptors agonists (e.g. TLR-7. TLR-8 and TLR-9), interleukins (e.g. IL2, IL4, IL7, IL15 and IL23), synthetic immunogen, and synthetic drug molecules (Rituximab). i) Method: Both TpA and an ssDNA are first modified chemically or labeled during manufacturing with chemical functionality to provide freely accessible chemical groups (e.g. functional groups) such as amine (-NH2) or carboxylic acids (-COOH)) to be used to create a chemical bond between ssDNA and TpA. EDO and NHS chemistry can be used. Similarly, IL proteins, synthetic immunogen, or synthetic drug molecules are covalently connected to ssDNA using EDO and NHS chemistry or Avidin-Biotin conjugation method. TpA includes a carboxylic acid reacts with the amine of the ssDNA to form TpA-ssDNA (Figure 5).

[0454] The TpA-ssDNA(l ’) is combined with ssDNA(1”)-GNR to form TpA-DNA(1)-GNR. ii) Method: Nucleic acid based TpA are directly conjugated to GNRs. A nucleic acid sequence (single strand) is synthesised such that a portion of it is complementary to a ssDNA having a thiol or disulfide group, which thiol or disulfide group can bind to GNRs. Under enzymatic or non-enzymatic conditions, H-S-DNA-TpA conjugate is formed through the formation of a double stranded nucleic acid as shown (Figure 6). Equimolar amounts of nucleic acid based TpA and ssDNA with the thiol or disulfide group are mixed in buffer solution such as Tris EDTA, T4 DNA ligase buffer from New England BioLabs, or similar buffer and is incubated for about 5 min to about 120 min at about 2°C to about 25 °C. For downstream application, H-S-ssDNA-TpA conjugate solution was used as is. This conjugate further binds with GNRs through thiol (or disulfide bond) to complete TpA-DNA-GNR.

[0455] 3) TTA-X1-GNR-ssDNA

[0456] A dilute solution of ssDNA (complementary to ssDNA conjugated to TpA) is added to one or more of the solutions A to C above of TTA-X1-GNR, such that its final concentration is about 0.01 ug / mL to about 1 .0 ug / mL and dilution of the solutions A to C above of TTA-X1- GNR is about 0.5 to about 10%. Depending upon the total volume of mixed solution, an incubation time of about 1 to about 24 hrs at about 2°C to about 25 °C. Excess ssDNA is removed using centrifugation. The mixed solution is centrifuged at about 5000 to about 20,000 ref for about 5 to about 15 minutes at a temperature of about 2°C to about 40 °C and the supernatant is replaced with Tis-EDTA buffer solution to form TTA-X1-GNR- ssDNA. (Figure 7)

[0457] 4) TTA-X1-GNR-X2-TpA i) Method 1 : A solution from 3) TTA-X1-GNR-ssDNA and ssDNA-TpA (from 2 i) above) are mixed in amounts such that the ssDNA of TTA-X1-GNR-ssDNA and the amount of the ssDNA from ssDNA-TpA are about equal. Excess ssDNA-TpA is removed using centrifugation and exchange of supernatant with Tis-EDTA buffer solution to form TTA-X1- GNR-X2-TpA. (Figure 8) ii) Method 2: A solution from 1) TTA-X1-GNR and H-S-DNA-TpA (from 2 ii) above) are mixed and is allowed to equilibrate for about 1 hr to about 24 hrs at about 2 °C to about 25 °C. Excess H-S-DNA-TpA is removed using centrifugation. The mixed solution is centrifuged at about 5000 to about 20,000 ref for about 5 to about 15 minutes at a temperature of about 2°C to about 40 °C and the supernatant is replaced with Tis-EDTA buffer solution to form TTA-X1-GNR-X2-TpA. (Figure 9). 5) Protecting TTA-X1-GNR-X2-TpA

[0458] Protecting the conjugate compound upon exposure to various elements, such as whole blood, are disclosed herein. Polymers can be used to protect the conjugate compound, such as synthetic polymers and / or natural polymers. Synthetic polymers are those suitable for use in mammals. Examples of synthetic polymers include PEG (polyethylene glycol), PVP (polyvinylpyrrolidone), PSS (polystyrene sulfonate), PNIPAM (poly(N-isopropylacrylamide)), ploxamer, diblock or triblock polymers, pH or thermo- responsive polymers, or a combination thereof. Natural polymers are those suitable for use in mammals. Examples of natural polymers include glycoproteins, albumin, gelatine, collagen, or a combination thereof. In addition to polymer(s) or separately, phospholipids (both low and high density), micelles, vesicles, liposomes, solid lipid nanoparticle (SLN), nanostructured lipid carriers (NLC), or a combination thereof.

[0459] For the protection of the conjugate compound with polymer(s), a selected polymer has a thiol or disulphide functional group (e.g. at a terminal end) or has been modified to include this functional group. A dilute solution of this polymer in TDW, organic solvent, or a combination thereof, is mixed with a solution of TTA-X1-GNR-X2-TpA. Excess polymer is removed using centrifugation. The mixed solution is centrifuged at about 5000 to about 20,000 ref for about 5 to about 15 minutes at a temperature of about 2 °C to about 40 °C and the supernatant is replaced with Tis-EDTA buffer solution to form a polymer wrapped TTA-X1-GNR-X2-TpA.

[0460] Nano-aggregates such as micelles, vesicles, liposomes, solid lipid nanoparticle (SLN), nanostructured lipid carriers (NLC) and / or supramolecular vesicles are described for encapsulation and / or surface adsorption of the conjugate compound for intratumoural injection. A solution of nano-aggregates (e.g. vesicles or liposome) in TDW, organic solvent, or a combination thereof, is formed with or without using an external energy source (e.g. ultrasound), solvent evaporation, or a combination thereof. This nano-aggregate solution is then concentrated to a particle concentration of about 2 to about 20 times higher, using centrifuge or dialysis, electrodialysis, Tangential Flow Filtration (TFF) or a combination thereof. This concentrated solution is then mixed with a polymer wrapped conjugate compound solution and is allowed to equilibrate for about 1 hr to about 24hrs at about 2°C to about 25 °C. Excess of the conjugate compound is removed using membrane filters, centrifuge, TFF, or a combination thereof.

[0461] 6) Examples of GNR-X2-TpA, TTA-X1-GNR, and TTA- X1-GNR-X2-TpA Conjugates i) With non-labile linker(s): ii) With labile linker(s):

[0462] A) TTA- X1-GNR-X2-TpA Conjugates with Non-Labile Linker(s)

[0463] In this method, an immunogenic compound conjugated to Fc portion (Fc tag), IgG 1 , lgG4 (e.g., PD1 inhibitor) was conjugated to GNRs (to give GNR-TpA) or TTA-GNR by converting amine groups (coming from lysine amino acids of Fc tag or the biomolecule) to thiol groups (-SH) and allowing spontaneous gold-sulfur bond formation. Therefore, this method can be extended to pre-conjugated Fc-IL4, Fc-IL7, Fc-IL9, FC-IL15 and FC-IL12 available from AcroBiosystem (or other supplier) and can produce TTA-GNR-TpA conjugate compounds. As this example method of conjugation relies on the presence of free amino acids in a biomolecule, it can also be extended to other immunogenic compounds that have one or more lysine amino acids available or are capable of being Fc tagged. In this method, in vitro testing using immunological tests (specifically, lateral flow assay techniques) using anti-species Ab (anti to Fc tag) and recombinant or extracted biological targets ( / .e., proteins, oligonucleotides, or small molecules) was used to prove conjugation via antigen specific binding (or in other words binding to target biomolecules). This showed visual detection of GNRs which absorb and emit light in the visible spectrum. As binding in confirmatory tests are immunological, these conjugates find and bind the intended target for the TTA (e.g., PSMA binding motif or Glu-urea-Lys hence forth PSMABM) and confirm the therapeutic molecule at desired treatment area (cancer tumor) in the intended use in humans.

[0464] Biomolecules (SATA (N-succinimidyl S-acetylthioacetate) chemistry, conversion of some or all amine groups, coming from lysine amino acids to thiol groups) were first activated for covalent binding to GNRs. However, any biofunctionalization strategy to append thiol to the biomolecule of interest produced the desired conjugate compounds. SATA chemistry enabled the synthesis of two half conjugates (PSMABM -G NR and GNR- IL2), which were tested separately for the success of conjugation through in vitro immunological binding before combining these two conjugates into one conjugate compound via systematic addition of SATA-activated IL2 / PSMABM to GNR solution followed by SATA-activated PSMABM / IL2 to GNR solution was performed. Confirmation of development of conjugate compound was achieved from immunological binding of conjugate compound to respective biologic on lateral flow assay stick. Similar experiments were performed to achieve PD1 and CD47 half conjugates to create PSMABM-GNR-PD1 , CD47-GNR-IL2 and CD47-GNR-PD1 . Conjugation of PSMABM to GNRs with SATA chemistry and further confirming the success of half conjugate and PSMABM -G NR- IL conjugate compound with LFA was achieved.

[0465] Lateral flow assays (LFAs) are rapid, paper-based in-vitro immunoassays commonly used to detect the presence or absence of target analytes in complex mixtures, and they are particularly useful for evaluating the functionality of biomolecule-gold nanoparticle conjugates. These tests typically consist of a sample pad, conjugate pad, nitrocellulose membrane, and absorbent pad assembled on a backing card. The conjugate pad contained GNRs conjugated to specific biomolecules such as antibodies, peptides, or small molecules. When a liquid sample was applied to the sample pad, it flowed laterally through the device by capillary action, rehydrating the conjugates which then bind to the target analyte if present. As the mixture flows further, it encountered test and control spots or lines on the nitrocellulose membrane containing immobilized capture molecules. Specific binding at these lines resulted in visible red or purple bands due to the color of the AuNPs, allowing for easy visual interpretation (See Figure 10).

[0466] In the context of studying biomolecule-GNR conjugates, LFAs served as a tool to assess the conjugation efficiency, binding activity, and stability of the functionalized nanoparticles. A successful test line / spot signal indicated that the conjugated biomolecules retained their affinity and functionality after being attached to the GNRs. Other immunoassay methods, such as ELISA, can also be used to showcase this binding.

[0467] In general, LFAs were spotted with 0.5 uL of analyte at a concentration of 1 mg / mL. Running buffer is 50 uL PBS with 1% T20. In some cases, 5 uL of 10% bovine serum albumin (BSA) was added to improve stability. Moreover, some biomolecules were found to best behave in a more native matrix. Specifically, PD1 lgG4 antibody conjugates were more stable when the running buffer solution was exchanged for a solution of bovine plasma.

[0468] The following non-labile linkers were made:

[0469] 1. GNR-S-Fc-IL

[0470] 2. GNR-S-PSMABM

[0471] 3. IL-Fc-S-GNR-S-PSMABM

[0472] 4. GNR-S-PD1

[0473] 5. GNR-S-CD47

[0474] 6. PD1-S-GNR-S-PSMABM

[0475] 7. IL-FC-S-GNR-S-CD47

[0476] 8. PD1-S-GNR-S-CD47 Material list:

[0477] Fc Tag interleukins (Fc-IL2 and Fc-IL15) proteins from AcroBiosystems. This supplier also supplies Fc-IL4, Fc-IL7, Fc-IL9, and FC-IL12 and as the Fc part for binding to GNRs, all of these are understood to conjugate like IL2.

[0478] Anti IL2 and Anti IL15 antibodies from Sino Biologies.

[0479] Recombinant IL15 protein from Sino Biologies.

[0480] PSMA binding motif (Glu-urea-Lys) (specific to human PSMA protein), synthesized at Sona.

[0481] Human PSMA protein from Antibodies Online

[0482] Human anti PSMA antibodies from Sigma-Aldrich™ a) GNR-X2-IL Conjugate

[0483] A pre-conjugated Interleukin with Fc portion of human antibody fragment (Fc-IL) offers, based on the known sequence, sufficient amine groups from lysine residues. Some or all of these amine groups were converted into thiol groups using SATA (N-succinimidyl S-acetylthioacetate) chemistry. Thiol activated Fc-IL protein were allowed to mix with GNRs to spontenously establish gold-sulfur bond for the development of a non-labile linker of GNR-Fc-IL half conjugate. Conjugation was determined from immunological binding of conjugate to anti-IL antibodies and / or anti species antibodies, ( / .e.,anti to host Fc Tag of IL2 / IL15). See Figure 11.

[0484] Method:

[0485] Step 1 : Dissolved IL2-Fc or IL15-FC at 0.1 to 5 mg / mL (e.g. 1 mg / mL) into TDW triply deionized water).

[0486] Step 2: Prepared a stock solution of SATA by dissolving it in DMF or DMSO at a concentration of 8 mg / mL and dilute it 10 times in TDW and label it as SATA solution.

[0487] Step 3: Added 1 uL of the SATA stock solution into 10 uL of IL2-Fc or IL15-Fc and incubated this solution for 30 min at room temperature.

[0488] Step 4: The sulfhydryl protecting group was removed just before GNR conjugation by first adding 3 uL hydroxylamine*HCI (0.5 M in TDW) to SATA modified IL and then incubating at room temperature for 1 h.

[0489] Step 5: Mixed thiolated IL2-Fc or IL15-FC, (0.1-5 ug / mL, e.g. 2.2 ug / mL was used) with GNRs (PEGylated GNRs and / or non-PEGylated GNRs) and incubated solution for (0.5-15 h, e.g. 1 hr) at 2-37 °C (e.g. room temperature).

[0490] To create PEGylated GNRs, GNRs were coated with polyethylene glycol polymer. For this 0.1-100 uL, e.g. 0.5 uL solution of thiol-functionalized polyethylene glycol, 5- 200kDa (e.g., 5kDa) (HS-PEG) of concentration 0.1 mg / mL was added for every 1 mL of conjugate.

[0491] Step 6: Conjugates were blocked with PEG (covered exposed GNRs surfaces after conjugation to prevent non-specific binding with intended target) and then spun at 2500- 100000 rpm, e.g. 12000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1%T20, 10% sucrose in 10mM PBS). Other buffers or detergents can be used.

[0492] Step 7: A UV-vis spectrum was then measured for each conjugate and starting GNRs before being tested via immunoassay sticks.

[0493] Step 8: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA). Visual detection of gold nanorods on a test spot or control spot was the confirmation of in vitro immunological binding between IL2 and anti IL2-Fc Ab.

[0494] A polyclonal Goat antihuman (GAH) antibody (Ab) was immobilized on nitrocellulose membrane as control spot and anti-l L2 antibody with Fc tag was immobilized on same nitrocellulose membrane as test spot. The Fc portion of IL2-Fc tagged antibody is a fragment of a humanized antibody and thus detects and binds to GAH. Even if there is no positive response on the test spot, due to the weaker binding between IL2 and anti-IL2 antibody, some conjugate was captured by the control spot to prove immunological binding. In this study, LFA was used; however, other immunoassay methods, such as ELISA, can be used to showcase this binding.

[0495] 1 . Lateral flow assay stick, made from nitro cellulose sheets, were spotted with Anti species (1 mg / mL GAH (goat anti human) in PBS, 0.5 uL spot volume) to create control spot and anti IL2-Fc or IL15-Fc Ab (1 mg / mL in PBS, 0.5 uL spot volume) to create test spots. The use of other spot volume of biologies concentration for test and control spot is possible. See Figure 10.

[0496] 2. Conjugate pads were spotted with GNR-IL2-FC or GNR-IL15-Fc conjugate.

[0497] 3. Sticks were developed using a running buffer (1% T20, 1% in PBS).

[0498] 4. Spots were compared and the results recorded (Table 1).

[0499] Results: The UV-vis trace (Figure 12: UV-VIS Scan of various GNR solutions) showed successful synthesis of GNR-IL2-FC. Similar results were obtained for GNR-IL15- Fc. LFA testing was presented in Figures 13a and 13b (Representative example of IL2 developed LFA sticks. Top two sticks showing immunological binding between anti species polyclonal Ab and Fc portion of the Fc-IL2 and the bottom picture showing addition immunological binding between Anti IL2 antibodies and IL2 part of GNR-IL2 conjugate) and results were concluded in Table 1 for GNR-IL12-Fc and similar results were for GNR-IL15- Fc.

[0500] Table 1

[0501] Conclusions:

[0502] 1 . Change in max peak position (2-8 nm) was indicative of IL2-Fc binging to GNRs as an absorption peak shift was always expected after a surface modification of gold nanoparticles with large molecules (> 1000 Da).

[0503] 2. In vitro testing worked (developed a visible control spot), Fc part of IL-Fc binded with GAH and stopped the flow of GNRs due to the presence of GNRs-Fc-IL2 structure. Therefore, a visible spot for control confirmedof GNRs- IL-Fc conjugation. Similarly, IL2 binds with Anti IL2 Ab of test spot and stopped the flow of GNRs, showing presence of GNRs-Fc-IL2 structure.

[0504] The method can be extended to other proteins that have Fc Tag conjugated or a protein that has lysine amino acids and hence, free amine groups. b) GNR-X2-PD1 Conjugate

[0505] PD1 antibodies are class lgG4 and have lysine residues, specifically at position 409 in the CH3 domain (Fc region of Ab) and thus offer many free amine groups, some (or all) of which can be converted to thiol for binding to GNRs.

[0506] Method:

[0507] Step l : Dissolved PD1 inhibitor at 0.1 to 5mg / ml (e.g. 1 mg / ml was used) into its dressing buffer.

[0508] Step 2: Prepared a stock solution of SATA by dissolving it in DMF or DMSO at a concentration of 8mg / ml and dilute it 10 times in TDW and label it as SATA solution. Step 3: Added 1 uL of the SATA stock solution into 10uL of PD1 solution and incubated sol for 30min at room temperature.

[0509] Step 4: Deprotected Sulfhydryl group (just before GNR-Ab conjugation) as follows

[0510] 1 . Added 1 uL hydroxylamine sol to 10uL of SATA modified PD1 .

[0511] 2. React this solution for 1 hrs at room temperature.

[0512] Step 5: Mixed thiolated PD1 (0.1-5ug / ml, e.g. 2.2ug / ml was used) with PEGylated GNRs (or non-PEGylated GNRs) and incubated solution for (0.5-15hrs, e.g. 1 hr was used) at 2-37 °C (e.g. room temperature).

[0513] Step 6: A portion of the above solution was tested for conjugation protection with polyethylene oxide. For this 0.1-1 OOuL, e.g. 0.5uL solution of thiol polyethylene oxide, 5kDa (HS-PEG) of concentration 0.1 mg / ml was added to 1 ml_ of conjugate.

[0514] Step 7: Spun conjugate and PEG protected conjugate solutions at 2500-100000 rpm, e.g. 12000 rpm for 1-60min, e.g. 10min was used. Discarded supernatant (all except 20-30ul) and dispersed GNRs in conjugate redispersion buffer (1%T20, 10% sucrose in 1XPBS). One vial from each set was used to test blocking with PEG (polyethylene glycol) polymer. Other buffers or detergents can be used.

[0515] Step 8: In-vitro Testing

[0516] As the goal for this GNR-PD1 conjugate was to add targeting conjugates i.e., PSMA or CD47 this conjugate was tested on LFA to ensure no binding to PSMA protein or CD47 protein. This ensured that when conjugate compounds are formed, a simple LFA will be diagnostic.

[0517] LFA Test 1 : PSMA protein was immobilized on a nitrocellulose membrane as test spot and anti-PSMA Ab was immobilized on a nitrocellulose membrane as control spot. This test was deemed successful as no binding between the GNR-PD1 conjugate and PSMA protein occurred. See Figure 14a.

[0518] LFA Test 2: A polyclonal Goat anti-human (GAH) antibody was immobilized on a nitrocellulose membrane as control spot and human CD47 (aa 18-135) recombinant protein was immobilized as a test spot. This test was deemed successful as no binding between the GNR-PD1 conjugate and CD47 protein occurred. As PD1 is a humanized antibody, GAH binding occurred. See Figure 14b. c) PSMABM-X1-GNR half conjugate

[0519] PSMABM is a dipeptide (Glu-urea-Lys) shown in Figure 15 and offer free amine group for SATA chemistry to convert the primary-amine of the side chain of lysine to thiol group (-SH). Thiol-functionalized PSMA forms gold-sulfur bonds with GNRs under suitable conditions.

[0520] Method: Step 1 : 295 uL of a SATA stock solution (0.8 mg / mL in 1 :10 DMSO / PBS) was added to 216 uL of PSMABM (1 mg / mL in PBS) and incubated it for 1 h at room temperature.

[0521] Step 2: The sulfhydryl protecting group was removed just before GNR conjugation by first adding 3 uL hydroxylamine*HCI (0.5 M in PBS) to SATA modified PSMABM and then incubating at room temperature for 1 h.

[0522] Step 3: SATA modified PSMABM was then added to GNRs or PEGylated GNRs and incubated for 1 h.

[0523] Step 4: Conjugates were then spun at 2500-100000 rpm, e.g. 12000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0524] Step 5: A UV-vis spectrum was then measured for each conjugate and starting GNRs before being tested via immunoassay sticks.

[0525] Step 6: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA).

[0526] Results:

[0527] UV-Vis: A change in position of peak between 800-900nm (LSPR, longitudinal surface plasmon resonance) for starting GNRs and conjugates can be seen in Figure 16 and is indicative of successful conjugation.

[0528] An LFA stick spotted with IL and a stick spotted with PSMA protein were tested for conjugates made from both type of GNRs. As it is clear in Figure 17, the conjugate is only binding (faint red spot) to PSMA protein and not to IL2. PSMABM with both type of GNRs, show successful binding to PSMA protein. Other chemistries can be used to achieve different loading levels and loading. d) CD47-X1-GNR Conjugate

[0529] Conjugation of anti-CD47 antibodies to the surface of gold nanoparticles for targeting integrin-associated protein (IAP), also known as CD47, in cancer therapy. CD47 is overexpressed in many tumors and acts as a “don’t eat me” signal that prevents phagocytosis by binding to SIRPa on macrophages. Gold nanoparticles with anti-CD47 antibodies can target cancer tumor or cancer cells. Anti-CD47 antibodies — like all monoclonal antibodies — contain numerous lysine residues, which are common sites for bioconjugation. SATA chemistry is a bioconjugation technique which can convert the primary-amines of the side chain of lysine residues to thiol groups (-SH). Thiol- functionalized anti-CD47 antibodies will form gold-sulfur bonds with GNRs under suitable conditions. See Figure 18.

[0530] Method:

[0531] Step 1 : A stock solution of SATA was prepared by first dissolving SATA in DMF or DMSO to achieve a concentration of 8 mg / mL before being diluted again 10 times using TDW.

[0532] Step 2: 0.1 to 10 uL (e.g., 1.5 uL) of the SATA stock solution was then added to 0.1 to 50 uL (e.g., 30 uL) of anti-human CD47 (1 mg / mL in PBS) and incubated it for 1 h at room temperature.

[0533] Step 3: The sulfhydryl protecting group was removed just before GNR conjugation by adding 0.1 to 10 (e.g., 3 uL) of hydroxylamine*HCI (0.5 M in PBS) to SATA-modified anti-human CD47 and then incubating at room temperature for 1 h.

[0534] Step 4: 1 to 50 uL (e.g., 32 uL) SATA-modified anti-human CD47 was then added to 1 to 50 mL (e.g., 12 mL) of GNRs and incubated for 1 h.

[0535] Step 5: The conjugate was then spun at 2500-100000 rpm, e.g. 8000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0536] Step 6: A UV-Vis spectrum was then measured for each conjugate and starting GNRs before being tested via immunoassay sticks.

[0537] Step 7: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA).

[0538] Results:

[0539] As the goal for CD47-GNR conjugate was to be a part of CD47-GNR-IL2 or CD47- GNR-PD1 conjugate, it was tested on LFA to ensure binding to CD47 protein but no binding to IL2 or PD1 . This ensured that when these conjugate compounds are formed, a simple LFA will be diagnostic.

[0540] LFA Test 1 : A polyclonal Goat anti-human (GAH) antibody was immobilized on nitrocellulose membrane as control spot and human CD47 (aa 18-135) recombinant protein was immobilized as a test spot. This test was deemed successful as binding between the CD47-GNR conjugate and CD47 protein occurred. As CD47 is humanized, GAH binding occurred. Very faint second spot from GAH and CD47 binding because most of conjugate is bounded to CD47 protein. See Figure 19a.

[0541] LFA Test 2: A polyclonal Goat anti-human (GAH) antibody (Ab) was immobilized on nitrocellulose membrane as control spot and anti-l L2 antibody with Fc tag was immobilized as a test spot. This test was deemed successful as no binding between the CD47-GNRs conjugates and anti-l L2 antibody occurred. As CD47 is humanized, GAH binding occurred. Very faint top spot on the stick on label side. See Figure 19b.

[0542] Conclusions:

[0543] It is clear from the above experiments that the CD47-GNR conjugate has demonstrated in vitro immunological binding with respective biomolecules, thus proving successful formation of CD47-GNR half-conjugate. e) PSMABM- X1-GNR- X2-IL Conjugate

[0544] This conjugate combined the targeting capabilities of PSMABM with the immunotherapeutic capabilities of an Interleukin. This conjugate was made by forming a PSMABM-GNR conjugate as previously described and then incubating this sample with SATA-modified IL (Fc tagged). Thiol-functionalized IL-Fc formed gold-sulfur bonds with PSMABM-GNRs under suitable conditions to form the desired PSMABM-GNR-IL conjugate compounds (Option 1). Conversely, GNR-IL can be used to form the desired PSMABM- GNR-IL conjugate compounds by incubation with SATA-modified PSMABM (Option 2). See Figure 20.

[0545] Method (Option 1):

[0546] Step 1 : A stock solution of SATA was prepared by first dissolving SATA in DMF or DMSO to achieve a concentration of 8 mg / mL before being diluted again 10 times using TDW.

[0547] Step 2: 10 to 500 (e.g., 295 uL) of the SATA stock solution was then added to10 to 500 (e.g., 216 uL) of PSMABM (1 mg / mL in PBS) and incubated it for 1 h at room temperature.

[0548] Step 3: The sulfhydryl protecting group was removed just before GNR conjugation by adding 0.1 to 10 (e.g., 3 uL) of hydroxylamine*HCI (0.5 M in PBS) to SATA-modified PSMABM and then incubating at room temperature for 1 h.

[0549] Step 4: 20 to 1000 (e.g., 511 uL) of SATA-modified PSMABM was then added to 1 to 50 (e.g., 12 mL) of GNRs and incubated for 1 h.

[0550] Step 5: To the above PSMABM-GNR solution, 1 to 50 (e.g., 30 uL) of SATA- modified IL2-Fc was added and incubated for 1 h at room temperature.

[0551] Step 8: The resulting conjugate was then spun at 2500-100000 rpm, e.g. 8000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1% T20, 10% sucrose in 10mM PBS). Other buffers or detergents can be used.

[0552] Step 9: A UV-vis spectrum was then measured for the final conjugate and compared to the starting GNRs and half-conjugate before being tested via immunoassay sticks. Step 10: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA).

[0553] Results:

[0554] As the goal for this PSMABM-GNR-IL2 conjugate compound is to provide both targeting and therapeutic motifs to GNRs, this conjugate was tested on LFA for binding to PSMA protein and also IL2 to ensure the conjugate compound was formed.

[0555] LFA Test 1 : PSMA protein was immobilized on nitrocellulose membrane as test spot and anti-PSMA Ab was immobilized on nitrocellulose membrane as control spot. This test was deemed successful as binding to PSMA protein was observed. A notable attribute here conjugate is that only immunological binding to PSMA protein was observed and not to anti-PSMA antibody. This is due to no specificity of PSMABM to anti-PSMA Ab and as such binding was not observed on the control spot. See Figure 21a.

[0556] LFA Test 2: A polyclonal Goat anti-human (GAH) antibody (Ab) was immobilized on nitrocellulose membrane as control spot and anti-l L2 antibody with Fc tag was immobilized as a test spot. This test was deemed successful as binding to anti-l L2 antibody was observed. As IL2 is humanized (Fc portion), GAH binding occurred. See Figure 21 b.

[0557] UV-Vis: A change in position of peak between 800-900 nm (LSPR, longitudinal surface plasmon resonance) for starting GNRs versus conjugates can be seen in Figure 22 and showed successful conjugation. Figure 22 shows UV-Vis traces comparing washed GNRs, half-conjugate and conjugate compounds for PSMABM-GNR-IL2. LSPR values: GNR = 842 nm; PSMABM-GNR = 842 nm; PSMABM-GNR-IL2 = 856 nm.

[0558] Conclusions:

[0559] It is clear from the above experiments that the PSMABM-GNR-IL2 conjugate has demonstrated in vitro immunological binding with respective biomolecules, thus showing successful formation of PSMABM-GNR-IL2 conjugates. This result is also supported by a shift in LSPR value.

[0560] It is clear from the above experiment explanation that PSMABM-GNR-IL2 conjugate is demonstrating in-vitro immunological binding with respective bio-molecules, showing the successful formation of PSMABM-GNR-IL2 conjugate.

[0561] Although, PEG has been utilized in the manufacturing of the conjugates herein and high level of resilience of the conjugates towards biological mediums (serum, plasma, blood, and fate in blood stream) is expected. Surface functionalized or wrapped GNRs in molecules such as, collagen, elastin, polylactic acid, phospholipids etc. would assist in resilience. These conjugates can be used for cancer treatment and will eliminate multiple injections. f) PSMABM-X1-GNR-X2-PD1 Conjugate

[0562] This conjugate combined the targeting capabilities of PSMABM with the PD1 blocking capabilities of PD1 antibodies. This conjugate was made by forming a PSMABM- GNR half-conjugate as previously described and then incubating this sample with SATA- modified PD1 antibodies. Thiol-functionalized PD1 antibodies formed gold-sulfur bonds with PSMABM-GNRs under suitable conditions to form the desired PSMABM-GNR-PD1 conjugate compound (Option 1). Conversely, GNR-PD1 can be used to form the desired PSMABM-GNR-PD1 conjugate compound by incubation with SATA-modified PSMA (Option 2). See Figure 23.

[0563] Method (Option 2):

[0564] Step 1 : A stock solution of SATA was prepared by first dissolving SATA in DMF or DMSO to achieve a concentration of 8 mg / mL before being diluted again 10 times using TDW.

[0565] Step 2: 0.1 to 10 (e.g., 1 uL) of SATA stock solution was then added to 0.1 to 50 uL (e.g., 10 uL) of PD1 lgG4 Ab (1 mg / mL in PBS) and this solution was incubated for 1 h at room temperature.

[0566] Step 3: The sulfhydryl protecting group was removed just before GNR conjugation by adding 0.1 to 50 (e.g., 1 uL) of hydroxylamine*HCI (0.5 M in PBS) to SATA-modified PD1 lgG4 Ab and then incubating at room temperature for 1 h.

[0567] Step 4: 1 to 50 uL (e.g., 12 uL) of SATA-modified PSMABM was then added to 1 to 50 mL (e.g., 12 mL) of GNRs and incubated for 1 h.

[0568] Step 5: To the GNR-PD1 solution, 20 to 1000 uL (e.g., 300 uL) of SATA-modified PSMA was added and incubated for 1 h at room temperature.

[0569] Step 8: The resulting conjugate was then spun at 2500-100000 rpm, e.g. 8000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1% T20, 10% sucrose in 10mM PBS). Other buffers or detergents can be used.

[0570] Step 9: A UV-vis spectrum was then measured for the final conjugate and compared to the starting GNRs and half-conjugate before being tested via immunoassay sticks.

[0571] Step 10: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA).

[0572] Results: As the goal for this PSMABM-GNR-PD1 conjugate was to provide both targeting and therapeutic motifs to GNRs, this conjugate was tested on LFA for binding to PSMA and also PD1 to ensure the conjugate compound was formed. PD1 binding was shown previously for GNR-PD1 .

[0573] LFA Test: PSMA protein was immobilized on nitrocellulose membrane as test spot and anti-PSMA Ab was immobilized on nitrocellulose membrane as control spot. This test was deemed successful as binding to PSMA protein was observed. A notable attribute here conjugate is that only immunological binding to PSMA protein was observed and not to anti-PSMA antibody. This is likely due to the very low to no specificity of PSMABM to anti-PSMA Ab and as such binding was not observed on the control spot. See Figure 24.

[0574] UV-Vis: A change in position of peak between 800-900 nm (LSPR, longitudinal surface plasmon resonance) for starting GNRs versus conjugates can be seen in Figure 25 and showed successful conjugation. Figure 25 showed UV-Vis traces comparing washed GNRs, half-conjugate and conjugate compound for PSMABM-GNR-PD1. LSPR values: GNR = 842 nm; GNR-PD1 = 842 nm; PSMABM-GNR-PD1 = 852 nm.

[0575] Conclusions:

[0576] It is clear from the above experiments that the PSMABM-GNR-PD1 conjugate has demonstrated in vitro immunological binding with respective biomolecules, thus showing formation of PSMABM-GNR-PD1 conjugates. This result is also supported by a shift in LSPR value. g) CD47-X1-GNR-X2-IL2 Conjugate

[0577] This conjugate combined the targeting capabilities of anti-CD47 antibodies with the immunotherapeutic capabilities of an Interleukin. This conjugate was made by forming CD47-GNR half-conjugate as previously described and then incubating this sample with SATA-modified IL (Fc tagged). Thiol-functionalized IL-Fc formed gold-sulfur bonds with CD47-GNRS under suitable conditions to form the desired CD47-GNR-IL conjugate compound (Option 1). Conversely, GNR-IL can be used to form the desired CD47-GNR- conjugate compound by incubation with SATA-modified anti-CD47 antibodies (Option 2). See Figure 26.

[0578] Method (Option 1):

[0579] Step 1 : A stock solution of SATA was prepared by first dissolving SATA in DMF or DMSO to achieve a concentration of 8 mg / mL before being diluted again 10 times using TDW.

[0580] Step 2: 0.1 to 10 (e.g e.g., 1.5 uL) of SATA stock solution was then added to 0.1 to 50 (e.g. e.g., 30 uL) of anti-CD47 antibodies (0.5 mg / mL in PBS) and this solution was incubated for 1 h at room temperature. Step 3: The sulfhydryl protecting group was removed just before GNR conjugation by adding 0.1 to 10 uL (e.g., 1 uL) of hydroxylamine*HCI (0.5 M in PBS) to SATA-modified anti-CD47 antibodies and then incubating at room temperature for 1 h.

[0581] Step 4: 1 to 50 uL (e.g., 30 uL) of SATA-modified anti-CD47 was then added to 1 to 50 mL (e.g., 12 mL) of GNRs and incubated for 1 h.

[0582] Step 5: To the CD47-GNR solution, 1 to 100 uL (e.g. e.g. 23 uL) of SATA-modified IL2 was added and incubated for 1 h at room temperature.

[0583] Step 8: The resulting conjugate was then spun at 2500-100000 rpm, e.g. 8000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1% T20, 10% sucrose in 10mM PBS). Other buffers or detergents can be used.

[0584] Step 9: A UV-vis spectrum was then measured for the final conjugate and compared to the starting GNRs and half-conjugate before being tested via immunoassay sticks.

[0585] Step 10: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA).

[0586] Results:

[0587] As the goal for this CD47-GNR-IL2 conjugate compound is to provide both targeting and therapeutic agents to GNRs. This conjugate was tested on LFA for binding to CD47 and also IL2 to ensure conjugate compound is formed.

[0588] LFA Test 1 : A polyclonal Goat anti-human (GAH) antibody was immobilized on nitrocellulose membrane as control spot and human CD47 (aa 18-135) recombinant protein was immobilized as a test spot. This test was deemed successful as binding between the CD47-GNR-IL2 conjugate and CD47 protein occurred. As CD47 and IL2 are humanized, GAH binding also occurred. See Figure 27a.

[0589] LFA Test 2: A polyclonal Goat anti-human (GAH) antibody (Ab) was immobilized on nitrocellulose membrane as control spot and anti-l L2 antibody with Fc tag was immobilized as a test spot. This test was deemed successful as binding to anti-l L2 antibody was observed. As IL2 is humanized (Fc portion), GAH binding occurred. See Figure 27b.

[0590] UV-Vis: A change in position of peak between 800-900 nm (LSPR, longitudinal surface plasmon resonance) for starting GNRs versus conjugates can be seen in Figure 28 and showed successful conjugation. Figure 28 showed UV-Vis traces comparing washed GNRs, half-conjugate and conjugate compound for CD47-GNR-IL2. LSPR values: GNR = 840 nm; CD47-GNR = 838 nm; CD47-GNR-IL2 = 850 nm.

[0591] Conclusions: It is clear from the above experiments that the CD47-GNR-IL2 conjugate has demonstrated in vitro immunological binding with respective biomolecules, thus showing formation of CD47-GNR-IL2 conjugates. This result is also supported by a shift in LSPR value. h) CD47- X1-GNR-X2-PD1 Conjugate

[0592] This conjugate combined the targeting capabilities of CD47 with therapeutic value from PD1 inhibitor (PD1 antibodies). This conjugate was made by forming a CD47-GNR half-conjugate as previously described and then incubating this sample with SATA- modified PD1 antibodies. Thiol-functionalized PD1 antibodies formed gold-sulfur bonds with CD47-GNRS under suitable conditions to form the desired CD47-GNR-PD1 conjugate compound (Option 1). Conversely, GNR-PD1 can be used to form the desired CD47-GNR- PD1 conjugate compound by incubation with SATA-modified CD47 (Option 2).

[0593] Method (Option 1):

[0594] Step 1 : A stock solution of SATA was prepared by first dissolving SATA in DMF or DMSO to achieve a concentration of 8 mg / mL before being diluted again 10 times using TDW.

[0595] Step 2: 0.1 to 10 uL (e.g., 1.5 uL) of SATA stock solution was then added to 0.1 to 50 uL (e.g., 30 uL) of anti-CD47 antibodies (0.5 mg / mL in PBS) and this solution was incubated for 1 h at room temperature.

[0596] Step 3: The sulfhydryl protecting group was removed just before GNR conjugation by adding 0.1 to 10 uL (e.g., 1 uL) of hydroxylamine*HCI (0.5 M in PBS) to SATA-modified anti-CD47 antibodies and then incubating at room temperature for 1 h.

[0597] Step 4: 1 to 50 uL (e.g., 30 uL) of SATA-modified anti-CD47 was then added to 1 to 50 mL (e.g., 12 mL) of GNRs and incubated for 1 h.

[0598] Step 5: To the CD47-GNR solution, 1 to 100 uL (e.g. e.g., 34 uL) of SATA-modified PD1 was added and incubated for 1 h at room temperature.

[0599] Step 8: The resulting conjugate was then spun at 2500-100000 rpm, e.g. 8000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1% T20, 10% sucrose in 10mM PBS). Other buffers or detergents can be used.

[0600] Step 9: A UV-vis spectrum was then measured for the final conjugate and compared to the starting GNRs and half-conjugate before being tested via immunoassay sticks.

[0601] Step 10: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA). Results:

[0602] As the goal for this CD47-GNR-PD1 conjugate was to provide both targeting and therapeutic values to GNRs, this conjugate was tested on immunoassay (LFA) for binding to CD47. Binding for PD1 was previously shown for GNR-PD1 via GAH.

[0603] LFA Test: A polyclonal Goat anti-human (GAH) antibody was immobilized on nitrocellulose membrane as control spot and human CD47 (aa 18-135) recombinant protein was immobilized as a test spot. This test was deemed successful as binding between the CD47-GNR-PD1 conjugate and CD47 protein occurred. As CD47 and PD1 are humanized, GAH binding also occurred. See Figure 30.

[0604] UV-Vis: A change in position of peak between 800-900 nm (LSPR, longitudinal surface plasmon resonance) for starting GNRs versus conjugates can be seen in Figure 31 and showed successful conjugation. Figure 31 showed UV-Vis traces comparing washed GNRs, half-conjugate conjugate compound for CD47-GNR-PD1 . LSPR values: GNR = 842 nm; CD47-GNR = 842 nm; CD47-GNR-PD1 = 840 nm.

[0605] Conclusions:

[0606] It is clear from the above experiments that the CD47-GNR-PD1 conjugate has demonstrated in vitro immunological binding with respective biomolecules, thus proving successful formation of CD47-GNR-PD1 conjugates. This result is also supported by a shift in LSPR value.

[0607] B) TTA- X1-GNR-X2-TpA Conjugates with Labile Linkers)

[0608] Efficacy for targeting, using PSMA and anti-CD47, as well as further functionalization of these molecules with drug conjugates for immunotherapy has been shown. Single strand DNA (ssDNA) or aptamers were provided to serve as labile linker(s).

[0609] To make half conjugates, two single-stranded DNA (ssDNA, aptamer) with complementary sequences were utilized. One ssDNA sequence was terminated by a carboxylic acid for bioconjugation (either at 3’ or 5’ end) to either TA or T pA and the other ssDNA sequence was terminated by a thiol functional group for gold-sulfur conjugation (at either at 3’ or 5’ end). Hybridization of these ssDNA stands formed double-stranded DNA (dsDNA), thus creating a cleavable covalent linker. IL2 and PD1 half-conjugates bound via dsDNA to GNR surfaces were formed. The PD1 half-conjugate was further conjugated with SATA-modified anti-CD47 to form the first conjugate compound with a cleavable drug conjugate. The same experimental conditions were used to create half conjugates using TA (PSMA and / or CD47 antibodies). Various configurations of conjugate compound can be achieved by interconnecting different half conjugates. i) GNR- X2-PD1 Conjugate Conjugation of PSMABM / CD47 antibodies to gold nanoparticles with a non-labile linker and conjugation of IL or PD1 via a labile linker enables targeted delivery to cancer tumor / cells in response to NIR light. PD1 antibodies contain primary amine groups (mainly on lysine residues), which can readily react with activated esters formed from ssDNA with carboxylic group using EDC / NHS chemistry. This made them suitable for stable covalent attachment to materials with carboxylic acid groups, enabling efficient and site-specific bioconjugation. Mercaptoethanol was used to quench excess EDC / NHS before adding PD1 antibodies. ssDNA was precipitated out of aqueous solution for purification, thus avoiding unwanted salts and EDC / NHS biproducts in later steps. See Figure 32.

[0610] Method:

[0611] Step 1 : 30 uL of 5’-COOH-(CH2)6-ssDNA (1 mg / mL solution in PBS) was dissolved in 0.5 mL of 0.1 M MES buffer (pH = 6.0).

[0612] Step 2: 0.8 uL of an EDC / sNHS solution (EDC 20 mM, sNHS 50 mM) in water was added and this solution was incubated for 30 mins at room temperature.

[0613] Step 3: Next, 3 uL of aPD1 lgG4 antibody 1 mg / mL solution was added before carefully increasing the pH of the solution to 7 to 8 using 0.1 M sodium hydroxide solution. This solution was then incubated for 1 h at room temperature

[0614] Step 4: 1 .7 uL of a 10 mM hydroxylamine*HCI solution was added, and the solution was incubated for 30 mins at room temperature.

[0615] Step 5: The solution was then dilute to 8 mL using 100% ethanol and then pelted via centrifuge by spinning 8x 1.5 mL Eppendorf tubes at 14000 rpm for 10 min. After spinning, solvent was pipetted off leaving behind ssDNA pellets. Eppendorf tubes were placed in a 37 °C oven for 30 min to completely dry before tubes were combined and dissolved in 0.5 mL PBS (7-8 pH).

[0616] Step 6: 30 uL of 5’-HS-(CH2)6-ssDNA (1 mg / mL solution in PBS) was added to the above solution and left to hybridize by incubating to 1 h at room temperature.

[0617] Step 7: The above solution of PD1-(CH2)6-dsDNA-(CH2)e-SH was then added to 8 mL of GNRs and incubated for 1 hour at room temperature. Next, 100 uL of 10% BSA was added to the solution and incubated for 30 mins to improve stability.

[0618] Step 8: The resulting conjugate was then spun at 2500-100000 rpm, e.g. 8000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0619] Step 9: A UV-Vis spectrum was then obtained and was compared to the UV-Vis spectrum starting GNRs before being tested via immunoassay sticks. Step 10: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA).

[0620] Results:

[0621] As the goal for this GNR-dsDNA-PD1 labile half-conjugate was to add targeting conjugates i.e., PSMA or CD47 therefore, this conjugate was tested on LFA no binding to PSMA protein or CD47 protein.

[0622] LFA Test 1 : PSMA protein was immobilized on a nitrocellulose membrane as test spot and anti-PSMA Ab was immobilized on a nitrocellulose membrane as control spot. This test was deemed successful as no binding between the GNR-dsDNA-PD1 conjugate and PSMA occurred. See Figure 33a.

[0623] LFA Test 2: A polyclonal Goat anti-human (GAH) antibody was immobilized on a nitrocellulose membrane (Figure 33b). Human CD47 (aa 18-135) recombinant protein was immobilized on a nitrocellulose membrane (Figure 33c). This test was deemed successful as no binding between the GNR-ds-DNA-PD1 conjugate and CD47 protein occurred. As PD1 is a humanized antibody, GAH binding occurred.

[0624] Conclusions:

[0625] It is clear from the above experiments that the labile GNR-ds-DNA-PD1 conjugate has demonstrated in vitro immunological binding with respective biomolecules, thus proving successful formation of GNR-ds-DNA-PD1 half-conjugate. j) GNR- X2-IL2 Conjugate

[0626] Conjugation of FclL2 was achieved much like for PD1 inhibitor. Fc tag of Fc-IL2 possess primary amine groups (primarily on lysine residues), which can efficiently react with activated esters generated via EDC / NHS chemistry. This facilitates stable covalent attachment to surfaces bearing carboxylic acid functionalities in this case ssDNA, enabling selective bioconjugation. Prior to antibody addition, excess EDC / NHS was quenched using mercaptoethanol to prevent unwanted side reactions. Purification of the ssDNA conjugates by ethanol precipitation removed residual salts and EDC / NHS biproducts, further streamlining downstream functionalization and preserving GNR performance. See Figure 34.

[0627] Method:

[0628] Step 1 : 30 uL of 5’-COOH-(CH2)6-ssDNA (1 mg / mL solution in PBS) was dissolved in 0.5 mL of 0.1 M MES buffer (pH = 6.0).

[0629] Step 2: 0.8 uL of an EDC / sNHS solution (EDC 20 mM, sNHS 50 mM) in water was added and this solution was incubated for 30 mins at room temperature. Step 3: Next, 3 uL of a 1 mg / mL IL2-Fc was added before carefully increasing the pH of the solution to 7 to 8 using 0.1 M sodium hydroxide solution. This solution was then incubated for 1 h at room temperature

[0630] Step 4: 1 .7 uL of a 10 mM hydroxylamine*HCI solution was added, and the solution was incubated for 30 mins at room temperature.

[0631] Step 5: The solution was then dilute to 8 mL using 100% ethanol and then pelted via centrifuge by spinning 8x 1.5 mL Eppendorf tubes at 14000 rpm for 10 min. After spinning, solvent was pipetted off leaving behind ssDNA pellets. Eppendorf tubes were placed in a 37 °C oven for 30 min to completely dry before tubes were combined and dissolved in 0.5 mL PBS (7-8 pH).

[0632] Step 6: 30 uL of 5’-HS-(CH2)6-ssDNA (1 mg / mL solution in PBS) was added to the above solution and left to hybridize by incubating to 1 h at room temperature.

[0633] Step 7: The above solution of IL2-(CH2)6-dsDNA-(CH2)6-SH was then added to 8 mL of GNRs and incubated for 1 hour at room temperature. Next, 100 uL of 10% BSA was added to the solution and incubated for 30 mins to improve stability.

[0634] Step 8: The resulting conjugate was then spun at 2500-100000 rpm, e.g. 8000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0635] Step 9: A UV-Vis spectrum was then obtained and was compared to the UV-Vis spectrum starting GNRs before being tested via immunoassay sticks.

[0636] Step 10: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA).

[0637] Results:

[0638] As the goal for this GNR-dsDNA-IL2 half-conjugate was to add targeting conjugates i.e., PSMA, this conjugate was tested on LFA to ensure no binding to PSMA protein and binding to IL2.

[0639] LFA Test 1 : PSMA protein was immobilized on a nitrocellulose membrane as test spot and anti-PSMA Ab was immobilized on nitrocellulose membrane as control spot. This test was deemed successful as no binding between the GNR- dsDNA-IL2 conjugate and PSMA occurred. See Figure 35a.

[0640] LFA Test 2: Anti-I L2 antibody was immobilized on a nitrocellulose membrane as a test spot and GAH antibodies were used as control. This test was deemed successful as binding between the GNR-dsDNA-IL2 conjugate and anti-l L2 antibody occurred along with binding between GNR-dsDNA-IL2 conjugate and GAH. As IL2-Fc is a humanized antibody, GAH binding was expected. See Figure 35b. UV-Vis: A change in position of peak between 800-900 nm (LSPR, longitudinal surface plasmon resonance) for starting GNRs versus conjugates can be seen in Figure 36 and showed successful conjugation. Figure 36 showed UV-Vis traces comparing GNR to GNR-dsDNA-IL2. LSPR values: GNR = 842 nm; GNR-ds-DNA-IL2 = 850 nm.

[0641] Conclusions:

[0642] It is clear from the above experiments that the labile GNR-ds-DNA-IL2 conjugate has demonstrated in vitro immunological binding with respective biomolecules, thus proving successful formation of GNR-ds-DNA-IL2 half-conjugate. This result is also supported by a shift in LSPR value. k) CD47- X1-GNR- X2-IL2 Conjugate

[0643] Either option below was used (See Figure 37)

[0644] Method (Option 1):

[0645] Step 1 : 30 uL of 5’-COOH-(CH2)6-ssDNA (1 mg / mL solution in PBS) was dissolved in 0.5 mL of 0.1 M MES buffer (pH = 6.0).

[0646] Step 2: 0.8 uL of an EDC / sNHS solution (EDC 20 mM, sNHS 50 mM) in water was added and this solution was incubated for 30 mins at room temperature.

[0647] Step 3: Next, 3 uL of aPD1 lgG4 antibody 1 mg / mL solution was added before carefully increasing the pH of the solution to 7 to 8 using 0.1 M sodium hydroxide solution. This solution was then incubated for 1 h at room temperature

[0648] Step 4: 1 .7 uL of a 10 mM hydroxylamine*HCI solution was added, and the solution was incubated for 30 mins at room temperature.

[0649] Step 5: The solution was then dilute to 8 mL using 100% ethanol and then pelted via centrifuge by spinning 8x 1.5 mL Eppendorf tubes at 14000 rpm for 10 min. After spinning, solvent was pipetted off leaving behind ssDNA pellets. Eppendorf tubes were placed in a 37 °C oven for 30 min to completely dry before tubes were combined and dissolved in 0.5 mL PBS (7-8 pH).

[0650] Step 6: 30 uL of 5’-HS-(CH2)6-ssDNA (1 mg / mL solution in PBS) was added to the above solution and left to hybridize by incubating to 1 h at room temperature.

[0651] Step 7: The above solution of PD1-(CH2)6-dsDNA-(CH2)6-SH was then added to 8 mL of GNRs and incubated for 1 h at room temperature.

[0652] Step 8: 20 uL of SATA-modified anti CD47 antibodies were then added to the above solution before incubating for 1 h at room temperature. Next, 100 uL of 10% BSA was added to the solution and incubated for 30 mins to improve stability.

[0653] Step 8: The resulting conjugate was then spun at 2500-100000 rpm, e.g. 8000 rpm for 1-60 min, e.g. 10 min. The supernatant, all except 20-30 uL, was then discarded and the pelleted GNRs were then dispersed in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0654] Step 9: A UV-Vis spectrum was then obtained and was compared to the UV-Vis spectrum starting GNRs before being tested via immunoassay sticks.

[0655] Step 10: In vitro conjugate testing was performed by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA).

[0656] Results:

[0657] As the goal for this CD47-GNR-dsDNA-PD1 conjugate compound is to provide both targeting and therapeutic motifs to GNRs. This conjugate was tested on LFA for binding to CD47. Binding for PD1 was previously shown for GNR-dsDNA-PD1 via GAH.

[0658] LFA test: A polyclonal Goat anti-human (GAH) antibody was immobilized on nitrocellulose membrane as control spot and human CD47 (aa 18-135) recombinant protein was immobilized as a test spot. This test was deemed successful as binding between the CD47-GNR-dsDNA-PD1 conjugate and CD47 protein occurred. As CD47 and PD1 are humanized, GAH binding also occurred. See Figure 38.

[0659] UV-Vis: A change in position of peak between 800-900 nm (LSPR, longitudinal surface plasmon resonance) for starting GNRs versus conjugates can be seen in Figure 39 and showed successful conjugation. Figure 39 showed UV-Vis traces comparing washed GNRs and labile conjugate compound for CD47-GNR-dsDNA-PD1. LSPR values: GNR = 842 nm; CD47-GNR-dsDNA-PD1 = 848 nm.

[0660] C) ADDITIONAL CONJUGATES

[0661] I) PSMABM-GNR-dsDNA-IL2 Conjugate

[0662] Method:

[0663] Step 1 : Prepare IL2-dsDNA-GNR conjugate as mentioned earlier.

[0664] Step 2: add 300 uL of SATA-modified anti PSMA binding motif to the above solution (make it as described earlier) and incubate for 1 h at room temperature. Next, add 100 uL of 10% BSA to the solution and incubated for 30 mins.

[0665] Step 3: Spin the resulting conjugate at 8000 rpm for 10 min. Remove all except 20- 30 uL of the supernatant and disperse pelleted GNRs in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0666] Step 4: obtain and compare UV-vis to starting GNRs to detect changes due to surface modification.

[0667] Step 5: Perform In vitro conjugate testing by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA). m) CD47-dsDNA-GNR-dsDNA-IL2 Conjugate

[0668] Method:

[0669] Step 1 : Prepare GNR-dsDNA-IL2 as described earlier.

[0670] Step 2: 30 uL of 5’-COOH-(CH2)6-ssDNA (1 mg / mL solution in PBS) was dissolved in 0.5 mL of 0.1 M MES buffer (pH = 6.0).

[0671] Step 3: 0.8 uL of an EDC / sNHS solution (EDC 20 mM, sNHS 50 mM) in water was added and this solution was incubated for 30 mins at room temperature.

[0672] Step 4: Next, 20uL of CD47 antibody (0.5 mg / mL solution) and carefully increase the pH of the solution to 7 to 8 using 0.1 M sodium hydroxide solution. Incubate this solution for 1 h at room temperature

[0673] Step 5: add 1.7 uL of a 10 mM hydroxylamine*HCI solution to above solution and incubate for 30 mins at room temperature.

[0674] Step 6: Dilute this solution to 8 mL using 100% ethanol and centrifuge by spinning 8x 1.5 mL Eppendorf tubes at 14000 rpm for 10 min. After spinning, pipette out solvent leaving behind ssDNA pellets. Place these Eppendorf tubes at 37 °C oven for 30 min to completely dry before tubes combining and dissolving in 0.5 mL PBS (7-8 pH).

[0675] Step 7: add 30 uL of 5’-HS-(CH2)e-ssDNA (1 mg / mL solution in PBS) to the above solution and leave it to hybridize by incubating to 1 h at room temperature to create CD47- dsDNA-SH.

[0676] Step 8: add 20 uL of above solution to GNR-dsDNA-IL2 conjugate and allow it to incubate for 1 hr.

[0677] Step 9: Spin the resulting conjugate compound at 8000 rpm for 10 min. Remove all except 20-30 uL of the supernatant and disperse pelleted GNRs in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0678] Step 10: obtain and compare UV-vis to starting GNRs to detect changes due to surface modification.

[0679] Step 11 : Perform In vitro conjugate testing by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA). n) PSMABM-GNR-dsDNA-PD1 Conjugate

[0680] Method:

[0681] Step 1 : Prepare PSMABM-GNR conjugate and prepare HS-dsDNA-PD1 as described earlier.

[0682] Step 2: Mix 530 uL of HS-dsDNA-PD1 solution with PSMABM-GNR conjugate and allow it to incubate for 1 hr.

[0683] Step 3: Spin the resulting conjugate compound at 8000 rpm for 10 min. Remove all except 20-30 uL of the supernatant and disperse pelleted GNRs in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS). Step 4: obtain and compare UV-vis to starting GNRs to detect changes due to surface modification.

[0684] Step 5: Perform In vitro conjugate testing by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA). o) CD47-dsDNA-GNR-dsDNA-PD1 Conjugate with both TTA-GNR and GNR- TPA linker as labile linkers

[0685] Method:

[0686] Step 1 : Create CD47-dsDNA-SH solution and GNR-dsDNA-PD1 conjugate as explained above.

[0687] Step 2: Mix 20 uL CD47-dsDNA-SH solution with 500uL GNR-dsDNA-PD1 conjugate and allow it to incubate for 1 hr.

[0688] Step 3: Spin the resulting conjugate compound at 8000 rpm for 10 min. Remove all except 20-30 uL of the supernatant and disperse pelleted GNRs in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0689] Step 4: obtain and compare UV-vis to starting GNRs to detect changes due to surface modification.

[0690] Step 5: Perform In vitro conjugate testing by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA). p) PSMABM-dsDNA-GNR-dsDNA-IL2 Conjugate Method:

[0691] Step 1 : Create GNR-dsDNA-IL2 conjugate as explained above.

[0692] Step 2: 30 uL of 5’-COOH-(CH2)6-ssDNA (1 mg / mL solution in PBS) was dissolved in 0.5 mL of 0.1 M MES buffer (pH = 6.0).

[0693] Step 3: 0.8 uL of EDC / sNHS solution (EDC 20 mM, sNHS 50 mM) in water was added and this solution was incubated for 30 mins at room temperature.

[0694] Step 4: Next, 300uL of PSMA binding motif (1 mg / mL solution) and carefully increase the pH of the solution to 7 to 8 using 0.1 M sodium hydroxide solution. Incubate this solution for 1 h at room temperature

[0695] Step 5: add 1.7 uL of a 10 mM hydroxylamine*HCI solution to above solution and incubate for 30 mins at room temperature.

[0696] Step 6: Dilute this solution to 8 mL using 100% ethanol and centrifuge by spinning 8x 1.5 mL Eppendorf tubes at 14000 rpm for 10 min. After spinning, pipette out solvent leaving behind ssDNA pellets. Place these Eppendorf tubes at 37 °C oven for 30 min to completely dry before tubes combining and dissolving in 0.5 mL PBS (7-8 pH).

[0697] Step 7: add 30 uL of 5’-HS-(CH2)6-ssDNA (1 mg / mL solution in PBS) to the above solution and leave it to hybridize by incubating to 1 h at room temperature to create PSMABM-dsDNA-SH. Step 8: Mix 20 uL PSMABM-dsDNA-SH solution with GNR-dsDNA-IL2 conjugate and allow it to incubate for 1 hr.

[0698] Step 9: Spin the resulting conjugate compound at 8000 rpm for 10 min. Remove all except 20-30 uL of the supernatant and disperse pelleted GNRs in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0699] Step 10: obtain and compare UV-vis to starting GNRs to detect changes due to surface modification.

[0700] Step 11 : Perform In vitro conjugate testing by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA). q) PSMABM-dsDNA-GNR-dsDNA-PD1 Conjugate with both TTA GNR and GNR-TPA linker as labile linker

[0701] Method:

[0702] Step 1 : Create PSMABM-dsDNA-SH solution and GNR-dsDNA-PD1 conjugate as explained above.

[0703] Step 2: Mix 20 uL CD47-dsDNA-SH solution with GNR-dsDNA-PD1 conjugate and allow it to incubate for 1 hr.

[0704] Step 3: Spin the resulting conjugate compound at 8000 rpm for 10 min. Remove all except 20-30 uL of the supernatant and disperse pelleted GNRs in conjugate redispersion buffer (1% T20, 10% sucrose in 1x PBS).

[0705] Step 4: obtain and compare UV-vis to starting GNRs to detect changes due to surface modification.

[0706] Step 5: Perform In vitro conjugate testing by using immunoassays, specifically chromatographic immunoassays, more specifically lateral flow immunoassays (LFA).

[0707] It will be understood that certain of the above-described structures, functions, and operations of the above-described embodiments are not necessary to practice the present invention and are included in the description simply for completeness of an exemplary embodiment or embodiments. In addition, it will be understood that specific structures, functions, and operations set forth in the above-described referenced patents and publications can be practiced in conjunction with the present invention, but they are not essential to its practice. It is therefore to be understood that the invention may be practiced otherwise than as specifically described without actually departing from the spirit and scope of the present invention as defined by the appended claims.

Claims

We Claim:

1. A conjugate compound comprising:TTA-X1-MNP-X2-TpA wherein:TTA is a tumour targeting agent;MNP is a metal nanoparticle;TpA is a therapeutic agent; andX1and X2are the same or different, and are each independently selected from a non-covalent linker or a covalent linker.

2. The conjugate compound of claim 1 , wherein at least one of X1and X2is a cleavable linker.

3. The conjugate compound of claim 1 or 2, wherein X1or X2is a cleavable linker.

4. The conjugate compound of any one of claims 1 to 3, wherein the cleavable linker is one or more of a chemical cleavable linker, enzyme cleavable linker, heat cleavable linker, pH cleavable linker, and a photochemical cleavable linker.

5. The conjugate compound of any one of claims 1 to 4, wherein the cleavable linker is a photochemical cleavable linker.

6. The conjugate compound of any one of claims 1 to 5, wherein the cleavable linker is cleavable in response to a stimulus.

7. The conjugate compound of any one of claims 1 to 6, wherein the stimulus comprises one or more of photo-irradiation, enzyme(s), nucleophilic / basic reagent(s), reducing agent(s), electrophilic / acidic reagent(s), organometallic and metal reagent(s), and oxidizing reagent(s), heat and pH change.

8. The conjugate compound of claim 7, wherein the stimulus is photo-irradiation.

9. The conjugate compound of any one of claims 1 to 8, wherein the cleavable linker is cleavable at a wavelength in UV, visible, and / or NIR region.

10. The conjugate compound of any one of claims 1 to 9, wherein the cleavable linker is cleavable at a wavelength of about 650 nm to about 1100 nm; about 700 nm to about1100 nm; about 750 nm to about 1100 nm; about 800 nm to about 1100 nm; about 850 nm to about 1100 nm; about 700 nm to about 1000 nm; about 750 nm to about 950 nm; about 800 nm to about 1000 nm; or about 900 nm to about 1000 nm.11 . The conjugate compound of any one of claims 1 to 10, wherein X1and X2are each independently selected from at least one base pair of nucleotides; at least two base pairs of nucleotides; at most 50 base pairs of nucleotides; at most 40 base pairs of nucleotides; at most 30 base pairs of nucleotides; at most 20 base pairs of nucleotides; or about 2 to about 20 base pairs of nucleotides.

12. The conjugate compound of any one of claims 1 to 11 , wherein X1and X2are each independently selected from a double strand of complementary polynucleotide or double strand of complementary oligonucleotide or a double strand of complementary DNA strands.

13. The conjugate compound of any one of claims 1 to 12, wherein X1and X2are each independently selected from DNA and / or RNA with a suitable sequence and length.

14. The conjugate compound of any one of claims 1 to 13, wherein X1and X2are each independently selected from a linker with a linking functional group (e.g. amide, esters, ethers, thioethers, thioester, dative bond, carbamate, Schiff base, secondary amine, hydrazone, oxime, diazo bond, isourea, isothiourea, sulfonamide, aryl amine, amidine, phosphoramidate, hydroxyl, alkene, alkyne, thiol, disulfide, etc.).

15. The conjugate compound of any one of claims 1 to 14, wherein X1and X2are each independently selected from valine-citruline, valine-alanine, or a combination of two to eight amino acids; a self-immolative unit (e.g., a PAB spacer, PEG spacer, etc.), and optionally hydrophilic groups (e.g., PEG); glucuronides, peptides, or a linker with a terminal maleimide group, hydrazone, and / or disulfide; and / or a non-covalent interaction such as hydrogen bonds.

16. The conjugate compound of any one of claims 1 to 15, wherein X1and X2are each independently selected from S or FcS.

17. The conjugate compound of any one of claims 1 to 16, wherein the MNP is a tube, rod, shell, cage, sphere, fiber, wire, stars, plate, sea archon, or a combination thereof.

18. The conjugate compound of any one of claims 1 to 17, wherein the MNP has an average particle size that is less than about 1 m in size, and typically from about 1 nm to about 900 nm in size and various ranges therebetween.

19. The conjugate compound of any one of claims 1 to 18, wherein the average particle size of the MNP is about 1 nm to about 100 nm.

20. The conjugate compound of any one of claims 1 to 19, wherein the MNP has one or more dimensions of the order of 100 nm or less.21 . The conjugate compound of any one of claims 1 to 20, wherein the MNP is a metal nanorod (MNR).

22. The conjugate compound of claim 21 , wherein the MNR ranges from about 1 to about 100 nm.

23. The conjugate compound of claim 21 or 22, wherein the MNR has a diameter or cross-section of between about 5 nm and about 50 nm, such as from about 5 nm, about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, or about 45 nm, to about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, about 45 nm, or about 50 nm.

24. The conjugate compound of any one of claims 21 to 23, wherein the MNR has an axial length of between about 20 nm and about 500 nm, such as from about 20 nm, about 30 nm, about 40 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 125 nm, about 150 nm, about 175 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, or about 450 nm, to about 30 nm, about 40 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 125 nm, about 150 nm, about 175 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, or about 500 nm.

25. The conjugate compound of any one of claims 21 to 24, wherein the MNR has an aspect ratio of from about 1.1 to about 100, about 1.1 , about 1.2, about 1.3, about 1.4, about 1.5, about 1 .6, about 1 .7, about 1 .8, about 1 .9, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, or about 90, to about 1 .2, about 1.3, about 1 .4, about 1.5, about1 .6, about 1 .7, about 1 .8, about 1 .9, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, or from about 1.1 to about 10.

26. The conjugate compound of any one of claims 21 to 25, wherein the MNR have a variance for the length, diameter, and / or aspect ratio of any given metal nanorod in a population or subpopulation can be at most 10%, 8%, 5%, 2%, 1% or 0.1% different from the average length, diameter, and / or aspect ratio for metal nanorods in the population.

27. The conjugate compound of any one of claims 21 to 26, wherein the MNR the population can be composed of at least 90%, 95%, 99% or 99.9% metal nanorods having a particular length, diameter, and / or aspect ratio.

28. The conjugate compound of any one of claims 21 to 27, wherein the metal is a transition metal, a precious metal, or a combination thereof.

29. The conjugate compound of any one of claims 1 to 28, wherein the metal is selected from gold, nickel, palladium, platinum, copper, silver, zinc, cadmium, or a combination thereof.

30. The conjugate compound of any one of claims 1 to 29, wherein the metal is gold.31 . The conjugate compound of any one of claims 1 to 30, wherein the MNP is a gold nanorod (GNR).

32. The conjugate compound of any one of claims 1 to 31 , wherein the MNP is capped with any suitable capping agents such as, and without being limited thereto, carboxylic acid, conventional citrate, and / or a positively charged ligand.

33. The conjugate compound of any one of claims 1 to 32, wherein the MNP is wrapped comprising a polymer (e.g. PEG).

34. The conjugate compound of any one of claims 1 to 33, wherein the TTA is an agent for recognizing a target cell.

35. The conjugate compound of any one of claims 1 to 34, wherein the compound accumulates in cancer (e.g. tumour) with the assistance from an EPR (enhanced permeability effect).

36. The conjugate compound of any one of claims 1 to 35, wherein the compound accumulates excessively in cancer (e.g. tumour) compared to healthy tissue.

37. The conjugate compound of any one of claims 1 to 36, wherein the TTA reduces toxicity from off-target delivery of the TpA.

38. The conjugate compound of any one of claims 1 to 37, wherein the target cell is a cancer cell.

39. The conjugate compound of any one of claims 1 to 38, wherein the target cell is a tumour cell.

40. The conjugate compound of any one of claims 1 to 39, wherein the TTA comprises a peptide, a protein, a polypeptide, an antibody or functional equivalent thereof, an antibody fragment, nucleic acids, nucleosides, aptamers, cell receptor inhibitors, hormones, or a combination thereof.41 . The conjugate compound of any one of claims 1 to 40, wherein the TTA comprises an antibody, or a functional equivalent thereof.

42. The conjugate compound of claim 41 , wherein the antibody or functional equivalent thereof is anti-CD20, anti-PSMA, PSMA binding motif (Glu-urea-Lys), anti-CD47, anti- EGFR or a combination thereof.

43. The conjugate of any one of claims 1 to 41 , wherein the TTA targets 1 GH-IGK, 43- 9F, 5T4, 791Tgp72, acyclophilin C-associated protein, alpha-fetoprotein (AFP), a-actinin-4, A3, antigen specific for A33 antibody, ART-4, B7, Ba 733, BAGE, BCMA, BCR-ABL, beta- catenin, beta-HCG, BrE3-antigen, BCA225, BTAA, CA125, CA 15-3\CA 27.29\BCAA, CA195, CA242, CA-50, CAM43, CAMEL, CAP- 1 , carbonic anhydrase IX, c-Met, CA19-9, CA72-4, CAM 17.1 , CASP-8 / m, CCCL19, CCCL21 , CD1 , CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, CD20, CD21 , CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD44, CD45, CD46, CD47, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD68, CD70, CD70L, CD74, CD79a, CD79b, CD80, CD83, CD95, CD126, CD132, CD133, CD138, CD147, CD154, CDC27, CDK4, CDK4m, CDKN2A, CO-029, CTLA4, CXCR4, CXCR7, CXCL12, HIF-1a, colon-specific antigen-p (CSAp), CEA (CEACAM5), CEACAM6, c-Met, DAM, E2A-PRL, EGFR, EGFRvlll, EGP-1 (TROP-2), EGP-2, ELF2-M, Ep-CAM, fibroblast growth factor (FGF), FGF-5, Flt-1 , Flt-3, folate receptor, G250 antigen, Ga733VEpCAM, GAGE, gp100, GRO- , H4-RET, HLA-DR, HM1.24, human chorionic gonadotropin (HCG) and its subunits, HER2 / neu, HMGB-1 , hypoxia inducible factor (HIF-1), HSP70-2M, HST-2, HTgp-175, la, IGF-1 R, IFN- y, IFN-a, IFN-P, IFN-A, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-2, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-23, IL-25, insulin-like growth factor-1 (IGF-1), KC4-antigen, KSA, KS- 1-antigen, KS1-4, LAGE-1a, Le-Y, LDR / FUT, M344, MA-50, macrophage migration inhibitory factor (MIF), MAGE, MAGE-1 , MAGE-3, MAGE-4, MAGE-5, MAGE-6, MART-1 , MART-2, TRAG-3, mCRP, MCP-1 , MIP-1A, MIP-1 B, MIF, MG7-Ag, M0V18, MUC1 , MUC2, MUC3, MUC4, MUC5ac, MUC13, MUC16, MUM-1 / 2, MUM-3, MYL-RAR, NB / 70K, Nm23H1 , NuMA, NCA66, NCA95, NCA90, NY-ESO-1 , p15, p16, p185erbB2, p180erbB3, PAM4 antigen, pancreatic cancer mucin, PD1 receptor (PD-1), PD-1 receptor ligand 1 (PD- L1), PD-1 receptor ligand 2 (PD-L2), PI5, placental growth factor, p53, PLAGL2, Pmel17 prostatic acid phosphatase, PSA, PRAME, PSMA, P1GF, ILGF, ILGF-1 R, IL-6, IL-25, RCAS1 , RS5, RAGE, RANTES, Ras, T101 , SAGE, S100, survivin, survivin-2B, SDDCAGi6, TA-90Mac2 binding protein, TAAL6, TAC, TAG-72, TLP, tenascin, TRAIL receptors, TRP-1 , TRP-2, TSP-180, TNF-a, Tn antigen, Thomson-Friedenreich antigens, tumor necrosis antigens, tyrosinase, VEGFR, ED-B fibronectin, WT-1 , 17-1A-antigen, complement factors C3, C3a, C3b, C5a, C5, an angiogenesis marker, bcl-2, bcl-6, K-ras, or any combination thereof.

44. The conjugate compound of any one of claims 1 to 43, wherein the T pA comprises an agent that treats cancer.

45. The conjugate compound of any one of claims 1 to 44, wherein the T pA comprises a protein, peptide, nucleic acid, amino acid, nucleoside, antibody, antibody drug conjugate(ADC), antibody fragment, antibody ligand, peptide nucleic acid, small organic molecule, lipid, hormone, drug, enzyme, lectin, cell adhesion molecule, antibody epitope, enzyme substrate, enzyme inhibitor, coenzyme, organic molecule, carbohydrate, such as polysaccharides and monosaccharides, or a combination thereof.

46. The conjugate compound of any one of claims 1 to 45, wherein the T pA comprises a toll like receptor (TLR) agonist, immunomodulator, drug molecule or a combination thereof.

47. The conjugate compound of claim 46, wherein the immunomodulator comprises a cytokine, cytokine agonists, a chemokine, chemokine agonists, a toll-like receptor (TLR) agonists, or a combination thereof.

48. The conjugate compound of claim 47, wherein the TLR agonist comprises a TLR-4 agonist, TLR-7 agonist, TLR-8 agonist, TLR-9 agonist, or a combination thereof.

49. The conjugate compound of any one of claims 46 to 48, wherein the immunomodulator comprises a cytokine.

50. The conjugate compound of claim 49, wherein the cytokine comprises IL-2, IL-4, IL- 6, IL-7, IL-8, IL-10, IL-12 IL-15, IL-17, IL-23, IL-1|3, TNF-a, IFN-a, IFN- , IFN-y, or a combination thereof.51 . The conjugate compound of any one of claims 46 to 50, wherein the immunomodulator comprises a chemokine, such as CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CXCL11 , CXCL12, CXCL14, CCL2, CCL5, or a combination thereof.

52. The conjugate of any one of claims 46 to 52, wherein the immunomodulator comprises a checkpoint inhibitor.

53. The conjugate compound of claim 52, wherein the checkpoint inhibitor comprises a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, or a combination thereof.

54. The conjugate compound of claim 53, wherein the checkpoint inhibitor comprises nivolumab, pembrolizumab, cemiplimab, ipilimumab, atezolizumab, avelumab, durvalumab, relatlimab, or a combination thereof.

55. The conjugate compound of any one of claims 1 to 54, wherein the T pA comprises a chemotherapeutic agent.

56. The conjugate compound of any one of claims 1 to 55, wherein T pA comprises Temozolomide, Actinomycin, Alitretinoin, All-trans retinoic acid, Azacitidine, Azathioprine,Bevacizumab, Bexatotene, Bleomycin, Bortezomib, Carboplatin, Capecitabine, Cetuximab, Cisplatin, Chlorambucil, Cyclophosphamide, Cytarabine, Daunorubicin, Docetaxel, Doxifluridine, Doxorubicin, Epirubicin, Epothilone, Erlotinib, Etoposide, Fluorouracil, Gefitinib, Gemcitabine, Hydroxyurea, Idarubicin, Imatinib, Ipilimumab, Irinotecan, Mechlorethamine, Melphalan, Mercaptopurine, Methotrexate, Mitoxantrone, Ocrelizumab, Ofatumumab, Oxaliplatin, Paclitaxel, Panitumab, Pemetrexed, Rituximab, Tafluposide, Teniposide, Tioguanine, Topotecan, Tretinoin, Valrubicin, Vemurafenib, Vinblastine, Vincristine, Vindesine, Vinorelbine, Vorinostat, Romidepsin, 5-fluorouracil (5-FU), 6- mercaptopurine (6-MP), Cladribine, Clofarabine, Floxuridine, Fludarabine, Pentostatin, Mitomycin, ixabepilone, Estramustine, prednisone, methylprednisolone, dexamethasone, or a combination thereof.

57. The conjugate compound of any one of claims 1 to 56, wherein the conjugate compound is wrapped with a polymer (e.g. PEG).

58. The conjugate compound of any one of claims 1 to 57, wherein the conjugate compound comprises PSMABM-X1-GNR-X2-IL2, CD47-X1-GNR-X2-IL2, PSMABM-X1-GNR- X2-PD1 , CD47-X1-GNR-X2-PD1 , or a combination thereof.

59. The conjugate compound of claim 58, wherein X1and X2are independently selected from S, ssDNA, dsDNA, or S-Fc.

60. The conjugate compound of claim 58 or 59, wherein the conjugate compound comprises PSMABM-GNR-IL2, CD47-GNR-IL2, PSMABM-GNR-PD1 , CD47-GNR-PD1 , PSMABM-S-GNR-SFCIL2, CD47-S-GNR-SFclL2, PSMABM-S-GNR-SFcPD1 , CD47-S- GNR-SFcPD1 , or a combination thereof.61 . The conjugate compound of any one of claims 1 to 60, wherein the conjugate compound comprises TTA-FcS-GNR-S-TpA, TTA-GNR-S-TpA, TTA-FcS-GNR-TpA, or a combination thereof.

62. The conjugate compound of any one of claims 1 to 61 , wherein the conjugate compound is capable of moving intratumorally.

63. The conjugate compound of any one of claims 1 to 62 for treatment of cancer.

64. The conjugate compound of claim 63, wherein the cancer is a tumour.

65. The conjugate compound of claim 64, wherein the cancer is a solid tumour.

66. The conjugate compound of any one of claims 1 to 65, for use in combination therapy, optionally, phototherapy, photothermaltherapy and immunotherapy.

67. A pharmaceutical composition comprising the conjugate compound of any one of claims 1 to 66.

68. The pharmaceutical composition of claim 67, wherein the composition is a dispersion, optionally, a colloidal dispersion.

69. The pharmaceutical composition of claim 67 or 68, further comprising pharmaceutically acceptable excipient(s).

70. The pharmaceutical composition according to any one of claims 67 to 69 for treatment of cancer.71 . The pharmaceutical composition of claim 70, wherein the cancer is a tumour.

72. The pharmaceutical composition of claim 71 , wherein the cancer is a solid tumour.

73. The pharmaceutical composition of any one of claims 67 to 72 for use in combination therapy, optionally, phototherapy, photothermaltherapy and immunotherapy.

74. A method for making the compound of any one of claims 1 to 66, the method comprising: combining TTA-CRG1 and CRG2-MNP-X2-TpA to make TTA-X1-MNP-X2-TpA, wherein CRG is a conjugate reactive group.

75. The method of claim 74, the method further comprises combining MNP-X2-TpA and CRG2 to make the CRG2-MNP-X2-TpA.

76. A method for making the compound of any one of claims 1 to 66, the method comprising: combining TTA-X1-MNP-CRG3 and CRG4-TpA to make TTA-X1-MNP-X2-TpA, wherein CRG3 and CRG4 are different, and are each selected to form X2, optionally, CRG4 is a portion of the TpA, wherein CRG is a conjugate reactive group.

77. The method of claim 76, wherein the method further comprises combining TTA-X1- MNP and CRG3 to make the TTA-X1-MNP-CRG3.

78. A method for making the compound of any one of claims 1 to 66, the method comprising: combining TTA-CRG1 , CRG2-MNP-CRG3, and CRG4-TpA to make TTA-X1-MNP- X2-TpA, wherein CRG is a conjugate reactive group.

79. The method of claim 78, wherein the method further comprises combining TTA- CRG1 , CRG2-MNP-CRG3, and CRG4-TpA, in any order.

80. The method of claim 78 or 79, wherein the method further comprises combining TTA-X1-MNP and CRG3 to make TTA-X1-MNP-CRG1 .81 . The method of any one of claims 74 to 80, wherein CRG1 and CRG2 are different, and are each selected from at least one nucleotide, forming at least one base pair of nucleotides.

82. The method of any one of claims 74 to 81 , wherein CRG1 and CRG2 are different, and are each selected from at least two nucleotides, forming at least two base pairs of nucleotides.

83. The method of any one of claims 74 to 82, wherein CRG3 and CRG4 are different, and are each selected from at least one nucleotide, forming at least one base pair of nucleotides.

84. The method of any one of claims 74 to 83, wherein CRG3 and CRG4 are different, and are each selected from at least two nucleotides, forming at least two base pairs of nucleotides.

85. The method of any one of claims 74 to 84, wherein the at least two nucleotides are polynucleotides (e.g. nucleic acids) or oligonucleotides.

86. The method of any one of claims 74 to 85, wherein CRG1 and CRG2 are each complement ssDNA.

87. The method of any one of claims 74 to 86, wherein CRG3 and CRG4 are each complement ssDNA.

88. The method of any one of claims 74 to 87, wherein CRG1 and CRG2 are each complement ssDNA that only bind to one another.

89. The method of any one of claims 74 to 88, wherein CRG3 and CRG4 are each complement ssDNA that only bind to one another.

90. The method of any one of claims 74 to 89, wherein CRG1 and CRG2 are different, and are each selected from a moiety or group capable of forming a covalent linker as a result of the association between atoms or molecules of each CRG1 and CRG2, forming a linking functional group.91 . The method of any one of claims 74 to 90, wherein CRG3 and CRG4 are different, and are each selected from a moiety or group capable of forming a covalent linker as a result of the association between atoms or molecules of each CRG3 and CRG4, forming a linking functional group.

92. The method of any one of claims 74 to 91 , wherein CRG1 to CRG4 are different, and are each selected from (a) carboxyl groups and various derivatives thereof including, but not limited to, N-hydroxysuccinimide esters (e.g. sulfo-N-hydroxysuccinimide), N-hydroxybenztriazole esters, acid halides, acyl imidazoles, thioesters, p-nitrophenyl esters, alkyl, alkenyl, alkynyl and aromatic esters; (b) hydroxyl groups which can be converted to esters, ethers, aldehydes, etc.; (c) haloalkyl groups wherein the halide can be later displaced with a nucleophilic group such as, for example, an amine, a carboxylate anion, thiol anion, carbanion, or an alkoxide ion, thereby resulting in the covalent attachment of a new group at the site of the halogen atom; (d) dienophile groups which are capable of participating in Diels-Alder reactions such as, for example, maleimido or maleimide groups; (e) aldehyde or ketone groups such that subsequent derivatization is possible via formation of carbonyl derivatives such as, for example, imines, hydrazones, semicarbazones or oximes, or via such mechanisms as Grignard addition or alkyllithium addition; (f) sulfonyl halide groups for subsequent reaction with amines, for example, to form sulfonamides; (g) thiol groups, which can be converted to disulfides, reacted with acyl halides, or bonded to metals such as gold, or react with maleimides; (h) amine or sulfhydryl groups (e.g., present in cysteine), which can be, for example, acylated, alkylated or oxidized; (i) alkenes, which can undergo, for example, cycloadditions, acylation, Michael addition, etc.; (j) epoxides, which can react with, for example, amines and hydroxyl compounds; (k) phosphoramidites and other standard functional groups useful in nucleic acid synthesis; (I) metal silicon oxide bonding; (m) metal bonding to reactive phosphorus groups (e.g., phosphines) to form, for example, phosphate diester bonds; (n) azides coupled to alkynes using copper catalyzed cycloaddition click chemistry; (o) biotin conjugate can react with avidin or strepavidin to form a avidin-biotin complex or streptavidin-biotin complex.

93. The method of any one of claims 74 to 92, wherein at least one of the CRGs are zero length linkers.

94. The method of any one of claims 74 to 93, further comprising adding a cross-linker between at least two CRGs.

95. The method of any one of claims 74 to 94, wherein TTA and / or TpA have at least one nucleotide to complement with CRG2 and CRG3, respectively.

96. A method for treating cancer, comprising administering to a mammal a therapeutically effective amount of the conjugate compound according to any one of claims 1 to 66 or the composition according to any one of claims 67 to 73.

97. The method of claim 96, wherein the conjugate compound or the composition targets the cancer (e.g. cancer cells).

98. The method of claim 96 or 97, wherein the TTA binds to the cancer cell.

99. The method of any one of claims 96 to 98, wherein the TTA binds to a tumour.

100. The method of any one of claims 96 to 99, wherein the TTA binds to a solid tumour.

101. The method of any one of claims 96 to 100, further comprising cleaving X1or X2.

102. The method of any one of claims claim 101 , wherein the cleaving comprises one or more of a chemical cleaving, enzyme cleaving, heat cleaving, pH cleaving, and a photochemical cleaving.

103. The method of claim 101 or 102, wherein cleaving comprises applying a stimulus to the conjugate compound or the composition.

104. The method of claim 103, wherein the stimulus comprises pH, enzymes, electromagnetic radiation (EMR), or a combination thereof.

105. The method of claim 104, wherein the stimulus comprises EMR.

106. The method of claim 104 or 105, wherein the EMR is selected from near-infrared, visible light, and UV, or a combination thereof.

107. The method of claim 106, wherein the EMR is near-infrared.

108. The method of any one of claims 104 to 107, wherein the EMR emits a wavelength in a range of about 700 nm to about 1000 nm, about 800 nm to about 1000 nm, about 900 nm to about 1000 nm, or about 950 nm to about 1000 nm.

109. The method of any one of claims 104 to 108, wherein the EMR excites the conjugate compound or the composition to generate heat.

110. The method of any one of claims 104 to 109, wherein the EMR cleaves X2to release the TpA.

111. The method of any one of claims 96 to 110, wherein the conjugate compound or composition generates heat to reduce growth of the tumour.

112. The method of any one of claims 104 to 111 , wherein the EMR is generated by a light-emitting diode (LED) or a laser generator.

113. The method of any one of claims 96 to 112, wherein the cancer is selected from brain tumour, nerve sheath tumour, breast cancer, colon cancer, colorectal cancer, skin cancer, lung cancer, hepatocellular cancer, gastric cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, cancer of the urinary tract, thyroid cancer, renal cancer, renal cell carcinoma, carcinoma, melanoma, lymphoma, head and neck cancer, brain cancer, and / or prostate cancer, including but not limited to androgendependent prostate cancer and androgen-independent prostate cancer.

114. The method of any one of claims 96 to 113, wherein the cancer is selected from prostate and / or breast cancer.

115. The method of any one of claims 96 to 114, wherein the cancer is a tumour.

116. The method of any one of claims 96 to 115, wherein the cancer is a solid tumour.

117. The method of any one of claims 96 to 116, administering the conjugate compound or composition as a combined therapy, optionally, photothermaltherapy and immunotherapy.

118. The method of any one of claims 96 to 117, wherein side-effects generated from off-target delivery and / or actions of the TpA is reduced.

119. The method of any one of claims 96 to 118, wherein the conjugate compound or the composition has a synergistic effect.

120. The method of any one of claims 96 to 119, wherein the conjugate compound or the composition has an additive effect.121 . The method of any one of claims 96 to 120, wherein the mammal is a human.

122. The method of any one of claims 96 to 121 , wherein administering comprises parenteral administration (e.g. formulated for injection via the intravenous, intramuscular, sub-cutaneous, intralesional, and / or intraperitoneal routes).

123. The method of any one of claims 96 to 122, wherein administering comprises a single injection for treatment versus two separate injections of TTA-MNP and TpA-MNP.

124. Use of a therapeutically effective amount of the conjugate compound according to any one of claims 1 to 66 or the composition according to any one of claims 67 to 73 for treating cancer.

125. The use of claim 124, wherein the conjugate compound or the composition targets the cancer (e.g. cancer cells).

126. The use of claim 124 or 125, wherein the TTA of the conjugate binds to the cancer cell.

127. The use of any one of claims 124 to 126, wherein the TTA binds to a tumour.

128. The use of any one of claims 124 to 127, wherein the TTA binds to a solid tumour.

129. The use of any one of claims 124 to 128, wherein X1or X2are cleavable.

130. The use of claim 129, wherein X1or X2are one or more of chemical cleavable, enzyme cleavable, heat cleavable, pH cleavable, and photochemical cleavable.

131. The use of any one of claims 124 to 130, wherein X1or X2are cleavable by application of a stimulus to the conjugate compound or the composition.

132. The use of claim 131 , wherein the stimulus comprises pH, enzymes, electromagnetic radiation (EMR), or a combination thereof.

133. The use of claim 132, wherein the stimulus comprises EMR.

134. The use of claim 132 or 133, wherein the EMR is selected from near-infrared, visible light, and UV, or a combination thereof.

135. The use of claim 134, wherein the EMR is near-infrared.

136. The use of any one of claims 132 to 135, a wavelength in a range of about 700 nm to about 1000 nm, about 800 nm to about 1000 nm, about 900 nm to about 1000 nm, or about 950 nm to about 1000 nm.

137. The use of any one of claims 132 to 136, wherein the EMR excites the conjugate compound or the composition to generate heat.

138. The use of any one of claims 132 to 137, wherein the EMR cleaves X2to release the TpA.

139. The use of any one of claims 124 to 138, wherein the conjugate compound or composition generates heat to reduce growth of the tumour.

140. The use of any one of claims 124 to 139, wherein the EMR is generated by a lightemitting diode (LED) or a laser generator.141 . The use of any one of claims 124 to 140, wherein the cancer is selected from brain tumour, nerve sheath tumour, breast cancer, colon cancer, colorectal cancer, skin cancer, lung cancer, hepatocellular cancer, gastric cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, cancer of the urinary tract, thyroid cancer, renal cancer, renal cell carcinoma, carcinoma, melanoma, lymphoma, head and neck cancer, brain cancer, and / or prostate cancer, including but not limited to androgendependent prostate cancer and androgen-independent prostate cancer.

142. The use of any one of claims 124 to 141 , wherein the cancer is selected from prostate and / or breast cancer.

143. The use of any one of claims 124 to 142, wherein the cancer is a tumour.

144. The use of any one of claims 124 to 143, wherein the cancer is a solid tumour.

145. The use of any one of claims 124 to 144, the conjugate compound or composition is used as a combined therapy, optionally, phototherapy and immunotherapy.

146. The use of any one of claims 124 to 145, wherein side-effects generated from off- target delivery and / or actions of the TpA is reduced.

147. The use of any one of claims 124 to 146, wherein the conjugate compound or composition has a synergistic effect.

148. The method of any one of claims 124 to 147, wherein the conjugate compound or the composition has an additive effect.

149. The use of any one of claims 124 to 148, wherein the mammal is a human.

150. The use of any one of claims 124 to 149, wherein the conjugate compound or composition is for parenteral administration (e.g. formulated for injection via the intravenous, intramuscular, sub-cutaneous, intralesional, and / or intraperitoneal routes).

151. The use of any one of claims 124 to 141 , wherein the conjugate compound or composition is a single injection for treatment versus two separate injections of TTA-MNP and TpA-MNP.

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