WO2020167868A1 - Gold nanoparticle-containing hydrogel films and chemotherapeutic methods for using same - Google Patents

Abstract

Disclosed are biocompatible implantable hydrogel films comprising a plurality of gold nanorods and at least one therapeutic or chemotherapeutic agent to facilitate a controlled, and localized hyperthermia within or about the body of an animal upon exposure to near-infrared laser energy. Also disclosed are methods for use of the gold nanorod-containing chemotherapeutic hydrogels in the treatment, prophylaxis, and/or the amelioration of one or more symptoms of mammalian cancer.

Description

DESCRIPTION

GOLD NANOPARTICLE-CONTAINING HYDROGEL FILMS AND CHEMOTHERAPEUTIC

METHODS FOR USING SAME

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority to United States Provisional Patent Application 62/803,776, filed February 11, 2019 (pending; Atty. Dkt. No.

37182.236PV01), the contents of which is specifically incorporated herein in its entirety by express reference thereto.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] Not Applicable. NAMES OF THE PARTIES TO A JOINT RESEARCH AGREEMENT

[0003] Not Applicable.

BACKGROUND OF THE INVENTION

FIELD OF THE INVENTION

[0004] The present disclosure relates to surgical devices and methods for surgical oncology. Disclosed are biocompatible films containing a plurality of gold nanorods and at least one therapeutic or chemotherapeutic agent to facilitate a controlled, and localized hyperthermia within or about the body of an animal, to which a near-infrared laser energy is applied. BRIEF DESCRIPTION OF THE INVENTION

[0005] The present invention overcomes these and other limitations inherent in the prior art by demonstrating the feasibility of a hydrogel film containing gold nanorods and a chemotherapeutic agent (which has been termed“Chemothermix film)” to produce localized controlled hyperthermia when treated with NIR laser.

[0006] Importantly, the disclosed hydrogen film provides application of the chemotherapeutic “payload” directly to the tissue surface (replacing, e.g., the use perfusion equipment for delivery of heated solubilized chemotherapy, which diffuses everywhere.)

[0007] Furthermore, unlike prior art products, the disclosed films produce a

"directed" hyperthermia, and are not simply a patch with uncontrolled release of a therapeutic agent.

[0008] By employing mild hyperthermia following implantation, an increased perfusion results in better uptake of the chemotherapeutic agents directly by the targeted tissues.

[0009] Because the disclosed films are also very thin and transparent, immediate visual feedback is available to the surgeon during implantation to facilitate better placement and localization of the film.

[0010] The data obtained from studies described herein suggest that the disclosed gold nanorod-embedded hydrogel films can be fabricated to contain selected chemotherapeutic agents, and these films can be precisely implanted during a surgical procedure to facilitate localized therapy that may maximize cell killing and reduce off- target toxicity. [0011] As shown herein, a Chemothermix film can be placed directly into the peritoneal cavity and mild hyperthermia can be facilitated following surgically-guided NIR laser irradiation to provide precision delivery and increase efficacy of chemotherapy to peritoneal malignancies.

[0012] This work enables the development of a new modality for treating not only peritoneal malignancies, but also any surgically-resected cancer.

[0013] The methods disclosed herein directly confront and advance the existing treatment paradigm for patients with peritoneal malignancies, and eliminate the need for perfusion equipment and its additional personnel to heat and circulate solubilized chemotherapy, while also enabling the delivery of chemotherapy in a more surgically precise and less toxic manner.

[0014] The new techniques can improve the safety of existing HIPEC (Hyperthermic IntraPEritoneal Chemotherapy) protocols by eliminating liters of solubilized chemotherapy currently used in the perfusion circuit, corresponding waste disposal costs and risk for dangerous spills. This invention also allows for more precise application of hyperthermia and chemotherapy to areas within the peritoneum containing microscopic tumors that cannot be resected ( e.g mesentery of bowel) and over areas at greatest risk of local recurrence (pelvis and right upper quadrant/right diaphragm). Moreover, gold nanorod-mediated hyperthermia provides more precise heating and can theoretically potentiate vascular permeability, much longer than current techniques, wherever the disclosed Chemothermix film is applied.

[0015] In one embodiment, the present disclosure provides a sterile, flexible, transparent, biologically compatible film that can be applied in the abdomen and other surgical sites to topically and directly deliver multimodal therapy to treat residual cancer and prevent local recurrence. The film’s hydrogel design allows it to conform and adhere to tissues, to remain in place where the surgeon applies it. This functionality— surgical application and hyperthermia in a targeted manner— is unique from other devices described in the prior art. Furthermore, the gold nanorods contained within the film can be precisely heated using NIR laser, with the goal of increasing the tumor vascular perfusion and improving the delivery of chemotherapy to tumors while synergistically increasing the killing efficiency of certain chemotherapies that have been shown to have increased efficacy in setting of hyperthermia¹. The design of the film allows for it to be cut to size and even to be applied in a layered fashion in areas to which the surgeon desires to increase the concentration of gold nanorods and chemotherapy. Its transparency allows the surgeon to visually monitor the effect of mild hyperthermia on the treatment area.

BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0017] The following drawings form part of the present specification and are included to demonstrate certain aspects of the present invention. For promoting an understanding of the principles of the invention, reference will now be made to the embodiments, or examples, illustrated in the drawings and specific language will be used to describe the same. It will, nevertheless be understood that no limitation of the scope of the invention is thereby intended. Any alterations and further modifications in the described embodiments, and any further applications of the principles of the invention as described herein are contemplated as would normally occur to one of ordinary skill in the art to which the invention relates.

[0018] The invention may be better understood by reference to the following description taken in conjunction with the accompanying drawings, in which like reference numerals identify like elements, and in which:

[0019] FIG. 1A and FIG. IB illustrate the conventional (FIG. 1A) and novel film- based (FIG. IB) methods of HIPEC. In FIG. 1A, chemotherapy is circulated by a roller pump, heated by a heat exchanger, and then delivered through large-bore tubing into the peritoneal cavity, under supervision of a perfusionist. Involves risk of spills and chemotherapy contamination; In FIG. IB, the surgeon applies film and NIR laser to deliver chemotherapy and heat in a targeted manner to achieve these exemplary results;

[0020] FIG. 2A, FIG. 2B, FIG. 2C, and FIG. 2D show various aspects of the present disclosure. FIG. 2A shows a non-functionalized film. Composition is Gelatin Type B (6% wt./vol.) and glycerol (25% wt./wt.). Thickness of 60 microns. Film is transparent, flexible, absorbs water quickly and, once wet, adheres on complex surfaces. FIG. 2B shows a Tumor-free mouse with l x l cm² film placed intraperitoneally. FIG. 2C shows the thermal imaging of tumor-free mouse with film placed intraperitoneally undergoing NIR laser treatment to generate hyperthermia. Temperature scale in degrees Celsius. NIR laser settings: Fluence 10 J/cm², Pulse frequency: lHz, Pulse width: Auto). FIG. 2D shows mild, controlled hyperthermia generated by NIR treatment of the disclosed film with gold nanorods (GNR). Non-survival surgery was performed on tumor-free mice. After laparotomy, mice were maintained at a temperature of 34-35°C using a heating pad (per IACUC guidelines), treated with NIR laser (Fluence 20 J/cm², Pulse width: 400 ms, Pulse frequency: lHz) for 60 seconds or until goal temperature of 42°C was reached, in 3 different experimental conditions: 1) No film, 2) Intraperitoneal Film without gold nanorods, or 3) Intraperitoneal Film with gold nanorods. The 1 x 1cm² films were placed either in the right upper quadrant (over the liver), left upper quadrant (over the stomach), or left lower quadrant (small and large intestines). The left lower and right lower quadrant are more uniform given the anatomic organ distribution in the murine abdomen. For each mouse, each type of film (including“absence of film”) was rotated to a different location to ensure the film location would not confound the results. The mean change in temperature from baseline was assessed using FLIR thermal imaging, error bars +/- 1 SE. Only the Au nanorod (GNR) film reached 42°C within 60 sec;

[0021] FIG. 3A and FIG. 3B show the results of elastic modulus analyses; In FIG. 3A, the logarithmic transformation of the elastic (Young’s) modulus of the 334- micron thick film at room temperature, human body temperature, and hydrated are shown; in FIG. 3B, the elastic modulus of the 334-micron thick film compared to other FDA- approved surgical materials is shown;

[0022] FIG. 4A and FIG. 4B show the transparency of 334-micron film with different shades of intensities. The raw image (FIG. 4A) and data analysis (FIG. 4B) is shown for each color type and intensity without (control) and with film;

[0023] FIG. 5A and FIG. 5B show NIR Irradiation Time and GNR Concentration Determines GNR-based Hyperthermia;

[0024] FIG. 6A, FIG. 6B, FIG. 6C, and FIG. 6D show non-functionalized films, and a surgical implantation procedure involving the

[0025] FIG. 7 shows Cisplatin is Efficiently Loaded and released from a hydrogen film; and

[0026] FIG. 8 shows a viability curve of various experimental conditions utilizing the compositions and methods disclosed herein. DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0027] Illustrative embodiments of the invention are described below. In the interest of clarity, not all features of an actual implementation are described in this specification. It will of course be appreciated that in the development of any such actual embodiment, numerous implementation-specific decisions must be made to achieve the developers’ specific goals, such as compliance with system-related and business-related constraints, which will vary from one implementation to another. Moreover, it will be appreciated that such a development effort might be complex and time-consuming but would be a routine undertaking for those of ordinary skill in the art having the benefit of this disclosure.

[0028] Peritoneal malignancies or cancers that spread to the peritoneum (lining of the abdomen) pose a treatment challenge. IV chemo often has minimal effect on these cancers, and surgical resection alone may miss microscopic tumors. Currently, recommended treatment is surgical removal of visible tumors followed by filling the abdomen with a heated chemo (fflPEC) bath to kill remaining cancer cells. This method has shown significant survival benefit for patients with metastatic gastric, colorectal, and ovarian cancers, with cures reported in patients with appendix cancer. Since the current fflPEC technique has a considerable recurrence risk and causes digestive dysfunction for up to a year after surgery, the present disclosure sought a re-invention of conventional fflPEC to make it more and less toxic.

[0029] Interestingly, tiny molecules of gold, known as nanoparticles, can generate heat by shining a laser on them. In the examples which follow, a film of gold nanoparticles was designed that included within it a chemotherapeutic that surgeons successfully placed into the abdomen, directly onto microscopic tumors. [0030] Following placement of the biocompatible film, the surgeon then shown a beam of laser light onto the film, which resulted in a localized heating of the target area. This delivered a localized chemotherapeutic dosing directly to maximize cancer cell killing, while at the same time, minimizing whole-body toxicity. This disclosure describes the particular properties of this film to provide controlled heating and to ensure the chemotherapeutic agent contained therein is efficiently released directly to the target site. Consequently, a new method of treatment for patients with peritoneal malignancies is now possible by allowing surgeons to directly apply chemotherapy to a particular diseased area without or about a surgical site.

[0031] The clinical benefits obtained from this invention are to improve chemotherapy efficacy and reduce toxicity for patients treated with film-based HIPEC. With improved efficacy, decreased risk of recurrence and not only increased disease-free survival but also overall survival for patients with peritoneal malignancies is now achievable. With decreased toxicity, an improved quality of life with decreased digestive dysfunction postoperatively can be expected as compared to the current standard-of-care surgical techniques. There are also multiple advantages of this work compared to the current methods of HIPEC. In addition to improving safety for operating room personnel²², this invention reduces costs through eliminating the need for the perfusion equipment and tubing (~$3500/case), need for the perfusionist, potentially decreasing operative time as heating and chemotherapeutic delivery will likely occur with less hands- on time than the current method of HIPEC, and also decreased cost by decreasing the operating time required for perfusion and its setup and breakdown. PERITONEAL MALIGNANCIES

[0032] Cancers that spread to or originate in the peritoneum— the surface lining of the abdominal cavity— are termed peritoneal malignancies. These cancers are often deemed unresectable and have limited treatment options, as chemotherapy administered intravascularly has little effect on these tumors and surgical resection may miss microscopic disease. In addition, these peritoneal tumors can cause small bowel obstructions, which prevent patients from eating, disqualify them from participating in clinical trials, and ultimately cause death. The standard of care is surgical removal of visible tumor (cytoreductive surgery or CRS) followed by infusion of heated chemotherapy into the abdominal cavity (HTPEC) (FIG. 1A). The advantage of HIPEC is controlled surface heating (~42°C) that is believed to facilitate drug penetration through increased tumor vascular perfusion^{1 3}.

[0033] Approximately 10,000 patients in the US are eligible for CRS/HIPEC each year^{4 6}. CRS/HIPEC has shown significant survival benefit in the treatment of primary and metastatic peritoneal malignancies, including colorectal, gastric, and ovarian cancers; peritoneal mesothelioma; and even cures for patients with appendix cancer. However, the current HIPEC technique uses an imprecise method of bathing the abdomen in solubilized chemotherapy (FIG. 1A). The existing approach has substantial risk of local recurrence and causes toxicity to intra-abdominal organs, manifested in patients as digestive dysfunction for up to a year after surgery⁷.

GOLD NANOPARTICLE-INDUCED HYPERTHERMIA

[0034] Gold nanoparticles, following excitement with near-infrared (NIR) laser radiation, can generate heat through surface plasmon resonance (SPR)⁸. SPR creates a photothermal effect when free electrons on the surface of the gold nanoparticle absorb NIR radiation of a specific wavelength, oscillate, and then produce heat. This unique property of gold nanoparticles has been exploited for ablative therapies (45°C). Phase I Clinical Trials have demonstrated a favorable safety profile of gold nanoparticles and NIR laser radiation in the ablative treatment of prostate cancer⁹ and are ongoing in head and neck cancer¹⁰. The feasibility of gold nanorods to generate controlled, localized heating to 42°C has recently been demonstrated¹¹. Gold nanorods administered intravascularly to mice with subcutaneous tumors provided precise mild hyperthermia within these tumors following near-infrared laser irradiation.

PREPARATION OF MEDICAMENTS

[0035] Another important aspect of the present invention concerns methods for using the disclosed Chemothermix films in the preparation of medicaments and devices for preventing, diagnosing, treating and/or ameliorating one or more symptoms of one or more diseases, dysfunctions, abnormal conditions, or disorders in an animal, including, for example, vertebrate mammals. Use of the disclosed Chemothermix film is particularly contemplated in the treatment of one or more abnormal conditions, such as the treatment of one or more cancer cell types in vivo, and surgically-resected cancers in particular. CHEMOTHERAPEUTIC METHODS AND USE

[0036] An important aspect of the present disclosure concerns methods for using the disclosed Chemothermix film for treating or ameliorating the symptoms of one or more forms of cancer, including, for example, a tumor or a metastatic cancer, such as, without limitation, peritoneal, appendiceal, and cancers of the digestive system. Such methods generally involve administering to a mammal (and in particular, to a human in need thereof), one or more of the disclosed Chemothermix film comprising a plurality of gold nanoparticles and at least a first anticancer therapeutic, in an amount and for a time sufficient to treat (or, alternatively ameliorate one or more symptoms of) the cancer in an affected mammal.

[0037] In certain embodiments, the Chemothermix film described herein may be provided to the animal in a single surgical intervention, or alternatively, in some embodiments, it may be desirable to repeat the treatment, or to include it in combination with one or more additional modes of therapy, for a period of several months or longer. In other embodiments, it may be desirable to provide the therapy in combination with one or more conventional HIPEC-based treatment regimens.

[0038] The present disclosure also provides for the use of one or more of the disclosed Chemothermix film in the manufacture of a medicament for therapy and/or for the amelioration of one or more symptoms of cancer, and particularly for use in the manufacture of a medicament for treating and/or ameliorating one or more symptoms of a mammalian cancer, including, for example human peritoneal cancers and the like.

THERAPEUTIC KITS

[0039] Therapeutic kits including one or more of the disclosed Chemothermix film and instructions for using the kit in a particular treatment modality also represent preferred aspects of the present disclosure. These kits may further optionally include one or more additional therapeutic compounds, one or more diagnostic reagents, or any combination thereof.

[0040] The kits of the invention may be packaged for commercial distribution, and may further optionally include one or more delivery devices adapted to deliver, localize, or secure the Chemothermix film to a selected target site within or about the body of an animal ( e.g ., syringes, sutures, surgical staples, meshes, injectables, implantable devices, and the like). Such kits typically include at least one vial, test tube, flask, bottle, syringe, or other container, into which the Chemothermix film(s) may be placed. Where a second pharmaceutical is also provided, the kit may also contain a second distinct container into which this second device or composition may be placed.

[0041] The kits of the present invention may also typically include a retention mechanism adapted to contain or retain the film(s) or other container(s) in close confinement for commercial sale, such as, e.g., injection or blow-molded plastic containers into which the desired film(s) or container(s) comprising them may be retained to minimize or prevent breakage, exposure to sunlight, or other undesirable factors, or to permit ready use of the composition(s) included within the kit. EXEMPLARY DEFINITIONS

[0042] In accordance with the present invention, polynucleotides, nucleic acid segments, nucleic acid sequences, and the like, include, but are not limited to, DNAs (including and not limited to genomic or extragenomic DNAs), genes, peptide nucleic acids (PNAs) RNAs (including, but not limited to, rRNAs, mRNAs and tRNAs), nucleosides, and suitable nucleic acid segments either obtained from natural sources, chemically synthesized, modified, or otherwise prepared or synthesized in whole or in part by the hand of man.

[0043] Unless defined otherwise, 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 invention belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Dictionary of Biochemistry and Molecular Biology , (2^nd Ed.) J. Stenesh (Ed ), Wiley-Interscience (1989); Dictionary of Microbiology and Molecular Biology (3^rd Ed.), P. Singleton and D. Sainsbury (Eds.), Wiley-Interscience (2007); Chambers Dictionary of Science and Technology (2^nd Ed ), P. Walker (Ed ),

Chambers (2007); Glossary of Genetics (5^th Ed.), R. Rieger et al. (Eds.), Springer-Verlag (1991); and The HarperCollins Dictionary of Biology, W.G. Hale and J.P. Margham, (Eds.), HarperCollins (1991).

[0044] Although any methods and compositions similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, and compositions are described herein. For purposes of the present invention, the following terms are defined below for sake of clarity and ease of reference:

[0045] In accordance with long standing patent law convention, the words“a” and “an,” when used throughout this application and in the claims, denote“one or more.”

[0046] The terms“about” and“approximately” as used herein, are interchangeable, and should generally be understood to refer to a range of numbers around a given number, as well as to all numbers in a recited range of numbers ( e.g “about 5 to 15” means“about 5 to about 15” unless otherwise stated). Moreover, all numerical ranges herein should be understood to include each whole integer within the range.

[0047] Biocompatible" refers to a material that, when exposed to living cells, will support an appropriate cellular activity of the cells without causing an undesirable effect in the cells, such as a change in a living cycle of the cells, a change in a proliferation rate of the cells, or a cytotoxic effect.

[0048] The term“biologically-functional equivalent” is well understood in the art, and is further defined in detail herein. Accordingly, sequences that have about 85% to about 90%; or more preferably, about 91% to about 95%; or even more preferably, about 96% to about 99%; of nucleotides that are identical or functionally-equivalent to one or more of the nucleotide sequences provided herein are particularly contemplated to be useful in the practice of the methods and compositions set forth in the instant application.

[0049] As used herein,“biomimetic” shall mean a resemblance of a synthesized material to a substance that occurs naturally in a human body and which is not rejected by (e.g, does not cause an adverse reaction in) the human body.

[0050] As used herein, the term“buffer” includes one or more compositions, or aqueous solutions thereof, that resist fluctuation in the pH when an acid or an alkali is added to the solution or composition that includes the buffer. This resistance to pH change is due to the buffering properties of such solutions, and may be a function of one or more specific compounds included in the composition. Thus, solutions or other compositions exhibiting buffering activity are referred to as buffers or buffer solutions. Buffers generally do not have an unlimited ability to maintain the pH of a solution or composition; rather, they are typically able to maintain the pH within certain ranges, for example from a pH of about 5 to 7.

[0051] As used herein, the term“carrier” is intended to include any solvent(s), dispersion medium, coating(s), diluent(s), buffer(s), isotonic agent(s), solution(s), suspension(s), colloid(s), inert(s) or such like, or a combination thereof, that is pharmaceutically acceptable for administration to the relevant animal. The use of one or more delivery vehicles for chemical compounds in general, and chemotherapeutics in particular, is well known to those of ordinary skill in the pharmaceutical arts. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the diagnostic, prophylactic, and therapeutic compositions is contemplated. One or more supplementary active ingredient(s) may also be incorporated into, or administered in association with, one or more of the disclosed chemotherapeutic compositions.

[0052] The term“effective amount,” as used herein, refers to an amount that is capable of treating or ameliorating a disease or condition or otherwise capable of producing an intended therapeutic effect.

[0053] The term“for example” or“e. ,” as used herein, is used merely by way of example, without limitation intended, and should not be construed as referring only those items explicitly enumerated in the specification.

[0054] As used herein, the phrase“in need of treatment” refers to a judgment made by a caregiver such as a physician or veterinarian that a patient requires (or will benefit in one or more ways) from treatment. Such judgment may made based on a variety of factors that are in the realm of a caregiver's expertise, and may include the knowledge that the patient is ill as the result of a disease state that is treatable by one or more compound or pharmaceutical compositions such as those set forth herein.

[0055] The phrases “isolated” or “biologically pure” refer to material that is substantially, or essentially, free from components that normally accompany the material as it is found in its native state.

[0056] As used herein, the term“kit” may be used to describe variations of the portable, self-contained enclosure that includes at least one set of reagents, components, or pharmaceutically-formulated compositions to conduct one or more of the assay methods of the present invention. Optionally, such kit may include one or more sets of instructions for use of the enclosed reagents, such as, for example, in a laboratory or clinical application. [0057] “Link” or “join” refers to any method known in the art for functionally connecting one or more proteins, peptides, nucleic acids, or polynucleotides, including, without limitation, recombinant fusion, covalent bonding, disulfide bonding, ionic bonding, hydrogen bonding, electrostatic bonding, and the like.

[0058] The term“naturally-occurring” as used herein as applied to an object refers to the fact that an object can be found in nature. For example, a polypeptide or polynucleotide sequence that is present in an organism (including viruses) that can be isolated from a source in nature and which has not been intentionally modified by the hand of man in a laboratory is naturally-occurring. As used herein, laboratory strains of rodents that may have been selectively bred according to classical genetics are considered naturally-occurring animals.

[0059] As used herein, the term“patient” (also interchangeably referred to as“host” or“subject”) refers to any host that can receive one or more of the pharmaceutical compositions disclosed herein. Preferably, the subject is a vertebrate animal, which is intended to denote any animal species (and preferably, a mammalian species such as a human being). In certain embodiments, a“patient” refers to any animal host including without limitation any mammalian host. Preferably, the term refers to any mammalian host, the latter including but not limited to, human and non-human primates, bovines, canines, caprines, cavines, corvines, epines, equines, felines, hircines, lapines, leporines, lupines, murines, ovines, porcines, ranines, racines, vulpines, and the like, including livestock, zoological specimens, exotics, as well as companion animals, pets, and any animal under the care of a veterinary practitioner. A patient can be of any age at which the patient is able to respond to inoculation with the present vaccine by generating an immune response. In particular embodiments, the mammalian patient is preferably human. [0060] The phrase“pharmaceutically-acceptable” refers to molecular entities and compositions that preferably do not produce an allergic or similar untoward reaction when administered to a mammal, and in particular, when administered to a human.

[0061] As used herein, “pharmaceutically acceptable salt” refers to a salt that preferably retains the desired biological activity of the parent compound and does not impart any undesired toxicological effects. Examples of such salts include, without limitation, acid addition salts formed with inorganic acids ( e.g hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like); and salts formed with organic acids including, without limitation, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic (embonic) acid, alginic acid, naphthoic acid, polyglutamic acid, naphthalenesulfonic acids, naphthalenedisulfonic acids, polygalacturonic acid; salts with polyvalent metal cations such as zinc, calcium, bismuth, barium, magnesium, aluminum, copper, cobalt, nickel, cadmium, and the like; salts formed with an organic cation formed from N, A/ -dibenzyl ethyl enedi a i ne or ethylenediamine; and combinations thereof.

[0062] As used herein, “polymer” means a chemical compound or mixture of compounds formed by polymerization and including repeating structural units. Polymers may be constructed in multiple forms and compositions or combinations of compositions.

[0063] As used herein, the term“polypeptide” is intended to encompass a singular

“polypeptide” as well as plural“polypeptides,” and includes any chain or chains of two or more amino acids. Thus, as used herein, terms including, but not limited to“peptide,” “dipeptide,” “tripeptide,” “protein,” “enzyme,” “amino acid chain,” and “contiguous amino acid sequence” are all encompassed within the definition of a“polypeptide,” and the term“polypeptide” can be used instead of, or interchangeably with, any of these terms. The term further includes polypeptides that have undergone one or more post-translational modification(s), including for example, but not limited to, glycosylation, acetylation, phosphorylation, amidation, derivatization, proteolytic cleavage, post-translation processing, or modification by inclusion of one or more non-naturally occurring amino acids. Conventional nomenclature exists in the art for polynucleotide and polypeptide structures.

[0064] For example, one-letter and three-letter abbreviations are widely employed to describe amino acids: Alanine (A; Ala), Arginine (R; Arg), Asparagine (N; Asn), Aspartic Acid (D; Asp), Cysteine (C; Cys), Glutamine (Q; Gin), Glutamic Acid (E; Glu), Glycine (G; Gly), Histidine (H; His), Isoleucine (I; He), Leucine (L; Leu), Methionine (M; Met), Phenylalanine (F; Phe), Proline (P; Pro), Serine (S; Ser), Threonine (T; Thr), Tryptophan (W; Trp), Tyrosine (Y; Tyr), Valine (V; Val), and Lysine (K; Lys). Amino acid residues described herein are preferred to be in the“L” isomeric form. However, residues in the “D” isomeric form may be substituted for any L-amino acid residue provided the desired properties of the polypeptide are retained.

[0065] As used herein, the terms“prevent,”“preventing,”“prevention,”“suppress,” “suppressing,” and“suppression” as used herein refer to administering a compound either alone or as contained in a pharmaceutical composition prior to the onset of clinical symptoms of a disease state so as to prevent any symptom, aspect or characteristic of the disease state. Such preventing and suppressing need not be absolute to be deemed medically useful.

[0066] “Protein” is used herein interchangeably with“peptide” and“polypeptide,” and includes both peptides and polypeptides produced synthetically, recombinantly, or in vitro and peptides and polypeptides expressed in vivo after nucleic acid sequences are administered into a host animal or human subject. The term“polypeptide” is preferably intended to refer to any amino acid chain length, including those of short peptides from about 2 to about 20 amino acid residues in length, oligopeptides from about 10 to about 100 amino acid residues in length, and longer polypeptides including from about 100 amino acid residues or more in length. Furthermore, the term is also intended to include enzymes, i.e., functional biomolecules including at least one amino acid polymer. Polypeptides and proteins of the present invention also include polypeptides and proteins that are or have been post-translationally modified and include any sugar or other derivative(s) or conjugate(s) added to the backbone amino acid chain.

[0067] “Purified,” as used herein, means separated from many other compounds or entities. A compound or entity may be partially purified, substantially purified, or pure. A compound or entity is considered pure when it is removed from substantially all other compounds or entities, i.e., is preferably at least about 90%, more preferably at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater than 99% pure. A partially or substantially purified compound or entity may be removed from at least 50%, at least 60%, at least 70%, or at least 80% of the material with which it is naturally found, e.g., cellular material such as cellular proteins and/or nucleic acids.

[0068] The term“subject,” as used herein, describes an organism, including mammals such as primates, to which treatment with the compositions according to the present invention can be provided. Mammalian species that can benefit from the disclosed methods of treatment include, but are not limited to, humans, non-human primates such as apes; chimpanzees; monkeys, and orangutans, domesticated animals, including dogs and cats, as well as livestock such as horses, cattle, pigs, sheep, and goats, or other mammalian species including, without limitation, mice, rats, guinea pigs, rabbits, hamsters, and the like.

[0069] As used herein,“synthetic” shall mean that the material is not of a human or animal origin.

[0070] The term“therapeutically-practical period” means the period of time that is necessary for one or more active agents to be therapeutically effective. The term "therapeutically-effective" refers to reduction in severity and/or frequency of one or more symptoms, elimination of one or more symptoms and/or underlying cause, prevention of the occurrence of symptoms and/or their underlying cause, and the improvement or a remediation of damage.

[0071] A“therapeutic agent” may be any physiologically or pharmacologically active substance that may produce a desired biological effect in a targeted site in a subject. The therapeutic agent may be a chemotherapeutic agent, an immunosuppressive agent, a cytokine, a cytotoxic agent, a nucleolytic compound, a radioactive isotope, a receptor, and a pro-drug activating enzyme, which may be naturally occurring, produced by synthetic or recombinant methods, or a combination thereof. Drugs that are affected by classical multidrug resistance, such as vinca alkaloids ( e.g ., vinblastine and vincristine), the anthracy clines (e.g., doxorubicin and daunorubicin), RNA transcription inhibitors (e.g, actinomycin-D) and microtubule stabilizing drugs (e.g, paclitaxel) may have particular utility as the therapeutic agent. Cytokines may be also used as the therapeutic agent. Examples of such cytokines are lymphokines, monokines, and traditional polypeptide hormones. A cancer chemotherapy agent may be a preferred therapeutic agent. For a more detailed description of anticancer agents and other therapeutic agents, those skilled in the art are referred to any number of instructive manuals including, but not limited to, the Physician's Desk Reference and Hardman and Limbird (2001).

[0072] “Treating” or“treatment of’ as used herein, refers to providing any type of medical or surgical management to a subject. Treating can include, but is not limited to, administering a composition comprising a therapeutic agent to a subject. “Treating” includes any administration or application of a compound or composition of the invention to a subject for purposes such as curing, reversing, alleviating, reducing the severity of, inhibiting the progression of, or reducing the likelihood of a disease, disorder, or condition or one or more symptoms or manifestations of a disease, disorder, or condition. In certain aspects, the compositions of the present invention may also be administered prophylactically, /. e. , before development of any symptom or manifestation of the condition, where such prophylaxis is warranted. Typically, in such cases, the subject will be one that has been diagnosed for being“at risk” of developing such a disease or disorder, either as a result of familial history, medical record, or the completion of one or more diagnostic or prognostic tests indicative of a propensity for subsequently developing such a disease or disorder.

[0073] The section headings used throughout are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application (including, but not limited to, patents, patent applications, articles, books, and treatises) are expressly incorporated herein in their entirety by express reference thereto. In the event that one or more of the incorporated literature and similar materials defines a term in a manner that contradicts the definition of that term in this application, this application controls. EXAMPLES

[0074] The following examples are included to demonstrate illustrative embodiments of the invention. It should be appreciated by those of ordinary skill in the art that the techniques disclosed in these examples represent techniques discovered to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of ordinary skill in the art should, in light of the present disclosure appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

EXAMPLE 1 - GOLD NANOROD-MEDIATED HYPERTHERMIA TO IMPROVE

CHEMOTHERAPEUTIC EFFICACY IN HIPEC

[0075] Disclosed is a flexible, transparent and biocompatible hydrogel film embedded with gold nanorods and 1) Mitomycin-C, 2) Cisplatin, or 3) Mitomycin-C and Cisplatin useful in delivering uniform hyperthermia and chemotherapy. Chemothermix film’s ability to heat uniformly in a controlled manner and to release chemotherapy has been characterized. The distribution of chemotherapy and gold nanorods in the film have been analyzed using the NIH Core facility at University of Washington for surface analysis using TOF-SIMS.

[0076] These findings may be verified through in vitro cell viability experiments using SNU16 (a human gastric cancer cell line) and HT-29 (a human colon cancer cell line) to ensure all positive ( e.g ., film with chemotherapy but without gold nanorods) and negative controls (e.g., film without chemotherapy or gold nanorods) behave as anticipated. Lastly, the distribution of gold nanorods following GMP-produced film application and NIR laser treatment in tumor-free mice (representing complete cytoreduction/CCO) and in murine models of 1) gastric cancer (SNU16) and 2) colon cancer (HT-29) with peritoneal metastases have been assessed and characterized.

[0077] In particular, this example describes the use of a Chemothermix film to produce controlled hyperthermia when treated with NIR laser (FIG. 2A, FIG. 2B, FIG. 2C, and FIG. 2D). The data suggest that a film containing gold nanorods and chemotherapy can deliver localized therapy that may maximize cell killing and reduce off- target toxicity. Chemothermix film has been fabricated to be placed directly into the peritoneal cavity, and by generating mild hyperthermia following surgically-guided NIR laser irradiation, precision delivery and increased efficacy of chemotherapy to peritoneal malignancies has been demonstrated (FIG. IB).

[0078] The resulting sterile, flexible, transparent, biologically compatible film can be applied not only in the abdomen, but also to other surgical sites to topically and directly deliver multimodal therapy to treat residual cancer and prevent local recurrence (FIG. 2A).

[0079] The disclosed film’s hydrogel design allows it to conform and adhere to tissues, to remain in place where the surgeon applies it. This functionality— surgical application and hyperthermia in a targeted manner— is unique from other devices described in the prior art. Furthermore, the gold nanorods contained within the film can be precisely heated using NIR laser (FIG. 2D), with the goal of increasing the tumor vascular perfusion and improving the delivery of chemotherapy to tumors while synergistically increasing the killing efficiency of certain chemotherapies that have been shown to have increased efficacy in setting of hyperthermia¹. The design of the film allows for it to be cut to size and even to be applied in a layered fashion in areas to which the surgeon desires to increase the concentration of gold nanorods and chemotherapy. Its transparency allows the surgeon to visually monitor the effect of mild hyperthermia on the treatment are a (FIG. 2B and FIG. 2C)

[0080] These films directly confront and advance the existing treatment paradigm for patients with peritoneal malignancies. The invention eliminates the need for perfusion equipment and its additional personnel to heat and circulate solubilized chemotherapy (FIG. 1A), while also enabling the delivery of chemotherapy in a more medically precise and less toxic manner. This technique also improves the safety of the HIPEC procedure by eliminating liters of solubilized chemotherapy currently used in the perfusion circuit, corresponding waste disposal costs and risk for dangerous spills. This invention also allows for more precise application of hyperthermia and chemotherapy to areas within the peritoneum containing microscopic tumors that cannot be resected ( e.g mesentery of bowel) and over areas at greatest risk of local recurrence (pelvis and right upper quadrant/right diaphragm) (FIG. IB). In addition, gold nanorod-mediated hyperthermia provides more precise heating and theoretically potentiate vascular permeability, much longer than current techniques, wherever the Chemothermix film is applied.

[0081] Although biocompatible films and chemotherapy have been used intraperitoneally for a few decades, the intraperitoneal administration of gold nanorods has not been well-studied in humans. However, other FDA-approved products exist with similar metals (palladium) embedded in a biocompatible material intended to remain in the abdominal cavity lifelong (Civasheet). The disclosed system should be safe for patients, especially given Phase I Clinical Trials that have demonstrated a favorable safety profile of gold nanoparticles and NIR laser radiation in ablative treatment of prostate⁹ and head and neck cancers¹⁰. Furthermore, regarding the composition of the Chemothermix film itself (gelatin and glycerol base), the safety of the concentration of glycerol used in the film was discussed with an expert in peritoneal dialysis who had even proposed its use as an osmotic agent (1.4%) in peritoneal dialysis. The use of this 1.5%(wt./vol.) glycerol- containing film in patients undergoing FQPEC should be as safe if not safer since it would have a significantly decreased dwell time and frequency in the peritoneum compared to peritoneal dialysate.

[0082] Quantifying In Vitro Release: Release of a chemotherapeutic agent from the Chemothermix film may be assessed by determining the concentration of the agent using high performance liquid chromatography (HPLC) in bench experiments of hydrated GMP- produced film in water. Such findings can also be confirmed in vitro utilizing cell viability (MTT assay) experiments in the gastric cancer cell line SNU16 and/or the colorectal cancer cell line FIT -29 to ensure that the non-functionalized film, the gold nanorod functionalized film, NIR laser radiation alone, and the mild hyperthermia generated from the gold nanorod functionalized film are not capable of killing cells in culture.

[0083] Likewise, exemplary films containing: 1) mitomycin-C, 2) cisplatin, or 3) a combination thereof (but not functionalized with gold nanorods), may be used to confirm that these agents delivered through a film platform are capable of decreasing cell viability.

[0084] To ensure uniform hyperthermia, the biodistribution of the gold nanorods can be determined when applied to the peritoneum of tumor-free Balb/c mice through a film platform, by functionalizing the film with fluorescently-tagged gold nanorods that can be visualized on the surface of the peritoneum and off-target organs using an In Vivo Imaging System (IVIS). This method of quantification will allow continuous monitoring of the distribution of gold nanorods in the mice over time (twice weekly) and is ultimately to assess biodistribution and toxicity at 8 timepoints from 24 hours to 3 months after film application (at the time of euthanasia). Furthermore, histopathologic analysis may also be employed to assess for abnormalities and TEM to quantify gold nanorod uptake in off- target organs.

[0085] The gold nanorods are expected to remain on the peritoneal surface with significantly less distribution into the parenchyma of off-target organs ( e.g liver, spleen, kidneys, and lungs), but if there is evidence of toxicity or deposition into off-target organs, the size and shape of gold nanoparticles may be altered to optimize biodistribution.

[0086] Gold nanorods delivered IV for treatment of a subcutaneous tumor can generate hyperthermia with a NIR irradiation approach and resulted in increased tumor perfusion and vascular permeability that persisted well beyond the period of NIR laser treatment¹¹. It is anticipated that this effect will also translate to the disclosed intraperitoneal film platform.

[0087] The Chemothermix film can be readily tested in murine models of gastric cancer and colon cancer with peritoneal metastases using the gastric cancer cell line SNU16 and colon cancer cell line HT-29, which have been stably transfected with a luciferase plasmid to generate luminescence. Mice may be monitored using IVIS until tumors are detectable in all four abdominal quadrants. Cytoreductive Surgery (CRS) need not be performed prior to film-based fflPEC, as would be done in surgical patients, so as to maintain measurable tumor for assessment of treatment response and permeability experiments. Mice may then undergo survival surgery with laparotomy and treatment with our GMP-produced Chemothermix film (and separate groups containing positive and negative control films), and subsequently treatment with near-infrared laser for 5-20 min; the duration of hyperthermia can be optimized from in vitro experiments. Temperature and perfusion in the peritoneal tumors can be measured using microprobes,¹¹ with half of each treatment group undergoing quantification of vascular permeability of peritoneal tumors through IV injection of Evans blue dye and subsequent euthanasia at 1, 3, and 24 hours, as previously described,¹¹ and the other half of each treatment group having twice- weekly blood collection to quantify the concentration of chemotherapeutic agents in plasma via HPLC.

[0088] Mice may also undergo IVIS imaging twice weekly to quantify peritoneal tumor burden. At 60 days after treatment or when mice have met euthanasia criteria, final IVIS imaging and blood collection can be performed. Following euthanasia, liver, spleen, lungs, and kidneys may be obtained for histopathologic analysis to assess toxicity and treatment effect, TEM for gold nanorod distribution, and HPLC to quantify chemotherapeutic concentration within off-target organs.

EXAMPLE 2 - PROCESS FOR PRODUCING A CHEMOTHERMIX FILM

[0089] This example describes the protocol for production of a hydrogel film composed of gelatin type B 6% and glycerol (from 1-25% glycerol) that is flexible, transparent, nontoxic, and in which gold nanorods (41 x 10 nm, ~4.1 ratio) (preferred concentration 2300 ppm AU to 4600 ppm Au) and chemotherapy, specifically Mitomycin- C (29.9 mM) and Cisplatin (333.3 mM) independently and in combination, can be embedded.

[0090] Chemotherapeutic concentrations are extrapolated to mice by maintaining concentrations consistent with those currently used for HIPEC in humans and accounting for proportional differences in body surface area between the two species.

MATERIALS REQUIREMENTS:

Soluble in water to be co-dissolved with water soluble chemotherapy drugs

High affinity with water - i.e. , tissue adhesive

Transparent

Flexible

Easy to cut and shape

Biocompatible

SOP for Chemothermix Film:

[0091] Set the temperature to 65°C on a stirrer with heating element.

[0092] Using a volumetric pipette, weigh 150 mg of glycerol in the glass beaker in which the solution will be prepared (Glycerol - Sigma Aldrich G5516).

[0093] Weigh 600 mg of gelatin in a weighing boat (Gelatin type B - Sigma Aldrich

G9382).

[0094] Add 10 mL of sterile water for injection (WFI) to the beaker with the glycerol.

[0095] Add gelatin to the beaker.

[0096] Add a magnetic stirring bar to the beaker.

[0097] Stir the solution at low speed at 65°C until all gelatin is dissolved, and a clear, yellow solution is achieved.

[0098] Remove the beaker from the stirrer and remove the stirring bar.

[0099] Turn off the heat to the stirrer. [0100] Add 10 pL of gold nanorods (Nanopartz stock solution 4.8 x 1013 nps/mL) and stir for additional 5 min.

[0101] Then add 299 pL of 1 mM stock solution of Mitomycin-C (or 3.33 pL of 1 M stock solution of Cisplatin or both chemotherapeutics) and stir for an additional 5 min.

[0102] Cast the gelatin/glycerol solution in the rectangular Teflon mold (6 mL/mold to yield an ~40-60-pm thickness film), being careful to avoid bubble formation.

[0103] Shake to coat the mold evenly.

[0104] Cover the mold and allow it to dry for 12 hrs.

[0105] Remove the film from the mold by carefully peeling off one side with a microforceps.

EXAMPLE 3 - CHEMOTHERMIX FILM CHARACTERIZATION

[0106] This example describes an exemplary biocompatible hydrogel film in accordance with the present invention that contains a plurality of gold nanorods and a solubilized chemotherapeutic that is gelatin-based, and includes glycerol for enhanced materials characteristics.

MATERIALS REQUIREMENTS:

[0107] Hydrogels of the present disclosure preferably are soluble in water to permit co-dissolution with water-soluble chemotherapy drugs; they preferably have a high affinity with water - tissue adhesive; are flexible, transparent, biocompatible, and easy to cut and shape to a selected size.

[0108] One such material as described herein has been characterized by staff of the Rice University Shared Equipment Authority, with testing performed on the ARES G2 Rheometer. The thickness of the film to maintain the desired materials characteristics varied typically from 28 microns to 334 microns.

[0109] The elasticity and tensile strength of the film was also tested at the same facility. The methodology used involved cutting the film into rectangular samples -3 x 15 mm that were then mounted without applied tension/compression and stretched with a Hencky strain rate of 10%/min. The logarithmic transformation of the elastic modulus of the 334-micron thickness film demonstrates that the film maintains its tensile strength even when used at human body temperatures. Also, the film becomes a compliant hydrogel with minimal tensile strength when it is hydrated. The elastic modulus of the 334-micron thickness film is similar to other surgical materials such as PTFE (vascular graft material) but is notably less than materials used primarily in the skin that require removal after wound healing such as Nylon, which is used in suture material.

[0110] The film transparency, in a situation mimicking film application to tissues, was estimated to be -90%. The results and visual analysis suggest that most tissue elements can be easily observed through the film at the moment of application (dry state).

Because the test may suffer signal pollution from reflection, the actual transparency is likely >95%. Upon contact with tissues and hydration, the transparency of the film is expected to approach 100%. The film transparency was estimated by capturing an image of gray- and red-scale patterns of different opacities (darkness) with and without the 334- micron thickness film overlying as is was expected that the thickest film would have the least transparency.

[0111] Images were analyzed by Fiji software in the following way: a rectangular sample area was selected over an individual bar of the desired opacity, histogram of pixel intensity was measured with“RGB weighted” option, and a mean intensity of the entire selected region was recorded. The measurement was performed for each opacity bar, and the data was recorded. The transparency (T%) was measured as T% = 100% * (If -Ib)/Ib, where If is the intensity of the bar with film and lb is the intensity of the bar without the film. It was observed that opacity-bars may be visually indistinguishable (printing artifacts) for opacities > 50%, thus the transparency was estimated only for the opacity range of 0-50% (FIG. 4B).

[0112] In additional embodiments, the film can be cast for one-directional drug release by making the film multilayer, i.e., casting sequentially layers of film with different chemical and/or mechanical composition. One layer can be used as a shield preventing drug elution to undesired tissue targets. This design feature can reduce the film’s chemotherapeutic concentration by increasing the exposure of the target tissue to drug. EXAMPLE 4 - SEPRAFILM CHARACTERIZATION

The disclosed biocompatible films have the following preferred characteristics:

• Attaches to tissues;

• Approved material;

• Easy procedure;

· Remains thin and easy to handle;

• Process involves water adsorption (compatible with solubilization of water soluble drugs); and

• Biocompatible: modified hyaluronic acid and carboxymethylcellulose. EXAMPLE 5 - SEPRAFILM WITHOUT GOLD NANORODS

[0113] Irradiated for <1 minute with temperature increase, but film without gold nanorods also had temperature increase; thus, need to optimize laser power: EXAMPLE 6 - LOCALIZED HYPERTHERMIA

[0114] The present invention generates localized hyperthermia upon exposure to near- infrared radiation. Off-target effects (represented by distilled water on left) and gold nanorod concentration have been optimized to minimize heating to surround structures that do not have film applied but may receive power scattered by laser delivery.

[0115] The images above demonstrate placement in the peritoneal cavity of a mouse with heating bar demonstrating goal temp of 42°C achieved.

[0116] Therapy for symptoms of mammalian disease or human cancer. Cisplatin (chemotherapy) is released from the film within 4 hours and can be quantified. Demonstrates cytotoxicity of chemotherapy (Cisplatin) released from the film and hyperthermia and from combination.

REFERENCES

[0117] The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein in their entirety by express reference thereto:

[0118] AHMED, S et al, “Outcomes with cytoreductive surgery and HIPEC for peritoneal metastasis,” J Surg. Oncol., 110(5):575-584 (2014).

[0119] ALTSCHUL, SF et al,“Gapped BLAST and PSI-BLAST: a new generation of protein database search programs,” Nucl. Acids Res., 25(17):3389-3402 (1997). [0120] CHOW, EK, and HO, D, “Cancer nanomedicine: from drug delivery to imaging,” Sci Transl Med., 5(216):216rv214 (2013).

[0121] CHUA, TC el al, “Early- and long-term outcome data of patients with pseudomyxoma peritonei from appendiceal origin treated by a strategy of cytoreductive surgery and hyperthermic intraperitoneal chemotherapy,” J. Clin. Oncol., 30(20):2449- 2456 (2012).

[0122] CONG, Y et al, “Alendronate-decorated biodegradable polymeric micelles for potential bone-targeted delivery of vancomycin,” J. Biomater. Sci. Polym. Ed., 26(ll):629-643 (2015).

[0123] DAKWAR, GR et al,“Nanomedicine-based intraperitoneal therapy for the treatment of peritoneal carcinomatosis - mission possible?” Adv. Drug Deliv. Rev., 108: 13-24 (2017).

[0124] DEDRICK, RL et al, “Pharmacokinetic rationale for peritoneal drug administration in the treatment of ovarian cancer,” Cancer Treat Rep., 62(1): 1-11 (1978).

[0125] DEHAL, A et al,“Cytoreductive surgery and intraperitoneal chemotherapy: an evidence-based review-past, present and future,” J. Gastrointest. Oncol., 7(1): 143-157 (2016).

[0126] GELLI, M et al, “Peritoneal and extraperitoneal relapse after previous curative treatment of peritoneal metastases from colorectal cancer: What survival can we expect?” Eur. J. Cancer (Oxford, England) 100:94-103 (Sep 2018).

[0127] GRIBSKOV, M, and BURGESS, RR,“Sigma factors from E. coli, B. subtilis, phage SP01, and phage T4 are homologous proteins,” Nucleic Acids Res., 14(16):6745- 6763 (Aug. 1986). [0128] HALE, WG, and MARGHAM, JP, “HARPER COLLINS DICTIONARY OF

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[0129] HARDMAN, JG, and LIMBIRD, LE, (Eds ),“GOODMAN AND GILMAN'S THE PHARMACOLOGICAL BASIS OF THERAPEUTICS” 10^th Edition, McGraw-Hill, New York (2001).

[0130] HINGORANI, SR et al. “HALO 202: Randomized Phase II Study of PEGPH20 plus Nab-Paclitaxel/Gemcitabine (PAG) Versus Nab-Paclitaxel/Gemcitabine in Patients with Untreated, Metastatic Pancreatic Ductal Adenocarcinoma” ./. Clin. Oncol., 36(4):359-366 (Dec. 2017).

[0131] HOLDER, A et al,“Operating rooms and equipment used for HIPEC are at risk for chemotherapy contamination,” 10th Int. Congr. Perit. Surf. Malignancies , 2016 Washington, DC.

[0132] HUANG, X, and EL-SAYED, MA, “Plasm onic photo-thermal therapy (PPTT )f Alexandria J. Med, 2011 47(1): 1-9.

[0133] JACQUET, P et al, “Hyperthermic intraperitoneal doxorubicin: pharmacokinetics, metabolism, and tissue distribution in a rat model,” Cancer Chemother. Pharmacol., 1998 41(2): 147-154.

[0134] KIRUI, DK et al, “Tumor vascular permeabilization using localized mild hyperthermia to improve macromolecule transport,” Nanomedicine, 2014 10(7): 1487- 1496.

[0135] KYTE, J, and DOOLITTLE, RF, “A simple method for displaying the hydropathic character of a protein,” J. Mol. Biol., 157(1): 105-132 (1982).

[0136] LOKESHWAR VB et al.“Targeting HA Family for cancer chemoprevention and therapy” Adv Cancer Research 123:35-65 (2014). [0137] LOPEZ-NORIEGA, A et al, “Hyperthermia-induced drug delivery from thermosensitive liposomes encapsulated in an injectable hydrogel for local chemotherapy,” Adv. Health. Mater., 3(6):854-859 (2014).

[0138] MCQUELLON, R et al., “Quality of life and nutritional assessment in peritoneal surface malignancy (PSM): recommendations for care,” J. Surg. Oncol.,

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[0140] OLIVA, A et al, “Development of an ultra high-performance liquid chromatography method for determining triamcinolone acetonide in hydrogels using the design of experiments/design space strategy in combination with process capability index,” J. Sep. Set, 2016 39(14):2689-2701.

[0141] PRECA, B-T et al,“A novel ZEB1/HAS2 positive feedback loop promotes EMT in breast cancer” Oncotarget, 8(7): 11530-43 (2017).

[0142] SHEN, H et al, “Chemotherapy sensitizes therapy-resistant cells to mild hyperthermia by suppressing heat shock protein 27 expression in triple negative breast cancer,” Clin. Cancer Res., DOI: 10.1158/1078-0432.CCR-17-3872, (Jun 2018).

[0143] SINGLETON, P and SAINSBURY, D,“DICHON ARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY ,” 2^nd Ed., John Wiley and Sons, New York (1987).

[0144] STERN, JM et al, “Initial evaluation of the safety of nanoshell-directed photothermal therapy in the treatment of prostate disease,” Int. J. Toxicol., 2016 35(1):38-

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[0146] Turley EA et al ., “Carcinoma cell hyaluronan as a ‘portable’ cancerized prometastatic microenvironment,” Cancer Res., 76(9):2507-2512, 2016.

[0147] URANO, M et al,“For the clinical application of thermochemotherapy given at mild temperatures,” Int. J Hyperthermia, 1999 15(2):79-107.

[0148] VISHNUBHAKTHULA, S et al,“Recent advances in hydrogel-based drug delivery for melanoma cancer therapy: A mini review,” J. Drug Deliv., 2017 2017:7275985.

[0149] It should be understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and the scope of the appended claims. All references, including publications, patent applications and patents, cited herein are hereby incorporated by reference to the same extent as if each reference was individually and specifically indicated to be incorporated by reference and was set forth in its entirety herein. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein.

[0150] The description herein of any aspect or embodiment of the invention using terms such as“comprising”,“having”,“including” or“containing” with reference to an element or elements is intended to provide support for a similar aspect or embodiment of the invention that“consists of’,“consists essentially of’, or“substantially comprises” that particular element or elements, unless otherwise stated or clearly contradicted by context (e.g, a composition described herein as comprising a particular element should be understood as also describing a composition consisting of that element, unless otherwise stated or clearly contradicted by context).

[0151] All of the compositions and methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents that are chemically and/or physiologically related may be substituted for the agents described herein while the same or similar results would be achieved.

[0152] All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

Claims

CLAIMS: WHAT IS CLAIMED IS:

1. A biocompatible gold nanorod-containing hydrogel film formed of a first layer of gelatin and glycerol, and having a plurality of gold nanorods, and at least one chemotherapeutic agent contained therein, and where in the first layer is from about 30 to about 300 pm thick.

2. The biocompatible gold nanorod-containing hydrogel of claim 1, comprising Type B gelatin.

3. The biocompatible gold nanorod-containing hydrogel of claim 1 or claim 2, further comprising a second distinct layer, the second layer differing in chemical, mechanical, solubility, or elution properties from the first layer.

4. The biocompatible gold nanorod-containing hydrogel of any preceding claim, wherein the gold nanorods are about 30-200nm in length

5. The biocompatible gold nanorod-containing hydrogel of any preceding claim, wherein the gold nanorods are about 5 to 50 nm in average diameter.

6. The biocompatible gold nanorod-containing hydrogel of any preceding claim, wherein the gold nanorods are about 40 nm in length and about 10 nm in diameter.

7. The biocompatible gold nanorod-containing hydrogel of any preceding claim, wherein the hydrogel has an average pore diameter of about 1 to about 50 microns.

8. The biocompatible gold nanorod-containing hydrogel in accordance with any preceding claim, wherein the at least one chemotherapeutic agent is selected from the group consisting of cyclophosphamide, doxorubicin, 5-fluorouracil, docetaxel, paclitaxel, trastuzumab, methotrexate, epirubicin, cisplatin, carboplatin, vinorelbine, capecitabine, gemcitabine, mitoxantrone, isabepilone, eribulin, lapatinib, carmustine, a nitrogen mustard, a sulfur mustard, a platin tetranitrate, vinblastine, etoposide, camptothecin, mitomycin C , and any combination thereof.

9. The biocompatible gold nanorod-containing hydrogel in accordance with any preceding claim, further comprising: a second, distinct chemotherapeutic agent, an immunomodulating agent, a neuroactive agent, an anti-inflammatory agent, an anti-lipidemic agent, a hormone, a receptor agonist, a receptor antagonist, an anti-infective agent, an antibody, an antigen-binding fragment of an antibody, a ribozyme, a cofactor, a steroid, or any combination thereof.

10. The biocompatible gold nanorod-containing hydrogel of any preceding claim, adapted and configured for surgical site implantation into a mammal.

11. The biocompatible gold nanorod-containing hydrogel in accordance with any preceding claim, formulated to generate a localized hyperthermia upon exposure to near-infrared radiation.

12. The biocompatible gold nanorod-containing hydrogel in accordance with any preceding claim, formulated for surgical site implantation following resection of a cancerous tumor, lesion, tissue, organ, or any combination thereof, in a human.

13. The biocompatible gold nanorod-containing hydrogel in accordance with any preceding claim, formulated for implantation following the resection of a cancerous tumor, lesion, tissue, or organ in the peritoneal cavity of a human, with between 1 and 40 films (covering 5 to 90% of the peritonealized surface) that are placed in or on the peritoneal cavity with total heating to 42 degrees Celsius of all films requiring from 30 seconds to 90 minutes.

14. The biocompatible gold nanorod-containing hydrogel in accordance with any preceding claim, adapted and configured as part of a therapeutic kit that comprises the hydrogel, and at least a first set of instructions for implantation of the hydrogel in the surgical site following resection of one or more cancerous cells or tissues within the human peritoneum.

15. The biocompatible gold nanorod-containing hydrogel in accordance with any preceding claim, for use in therapy, prophylaxis, or amelioration of one or more symptoms of a mammalian disease, disorder, dysfunction, deficiency, defect, trauma, injury, or abnormal condition.

16. The biocompatible gold nanorod-containing hydrogel in accordance with any preceding claim, for use in the therapy, prophylaxis, or amelioration of one or more symptoms of human cancer.

17. Use of a biocompatible gold nanorod-containing hydrogel in accordance with any one of claims 1 to 16, in the manufacture of an implant for treating or ameliorating at least one symptom of cancer in a mammalian subject.

18. Use in accordance with claim 79, wherein the mammalian subject is a human, a non-human primate, a companion animal, an exotic animal, a zoological specimen, or domesticated livestock.

19. A chemotherapeutic kit comprising: 1) a biocompatible gold nanorod-containing hydrogel in accordance with any one of claims 1 to 16; and 2) instructions for implantation of the hydrogel within the body a mammal following surgical resection of a tumor or a cancerous lesion.

20. A method of treating or ameliorating one or more symptoms of cancer in an animal, the method comprising administering to an animal in need thereof an effective amount of a biocompatible gold nanorod-containing hydrogel in accordance with any one of claims 1 to 16, for a time sufficient to treat or ameliorate the one or more symptoms of the cancer in the animal.

21 The method in accordance with claim 20, wherein the animal is human.

22 A method of administering a therapeutic or prophylactic agent to one or more cells, tissues, organs, or systems of a mammalian subject in need thereof, comprising surgically implanting into the subject an effective amount of the biocompatible gold nanorod-containing hydrogel in accordance with any one of claims 1 to 16.

23. The biocompatible gold nanorod-containing hydrogel of any one of claims 1 to 16, wherein the film is at least 50% transparent.

24. The biocompatible gold nanorod-containing hydrogel of claim 23, wherein the film is at least 90% transparent.

25. The biocompatible gold nanorod-containing hydrogel of any one of claims 1 to 16, having an elastic modulus of about 250-2500 MPa.