Compositions of chitosan and modified chitosans

Abstract

The present disclosure relates to pharmaceutical formulations comprising chitosan or modified chitosan. The present disclosure also relates to glycated chitosans and to methods of enhancing immune system response to an antigen comprising administration of glycated chitosans and / or their pharmaceutical formulations.

Description

[0001] Docket No.: IPW-00625

[0002] COMPOSITIONS OF CHITOSAN AND MODIFIED CHITOSANS

[0003] RELATED APPLICATIONS

[0004] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 730,254, filed December 10, 2024, the contents of which are incorporated herein by reference in their entirety.

[0005] BACKGROUND

[0006] Immunotherapy has been used with some success for systemic treatment of chemotherapy -resistant cancers. Cancer immunotherapy is based on inducing or boosting an immune response against the tumor(s). Specific methodologies to accomplish this have so far included cancer vaccines, modulation of immunity by cytokines or antibodies, adoptive transfer of immune cells, and others.

[0007] One such methodology to induce a robust and prolonged anti-tumor response (and hence effective overall clinical outcome) is to combine tumor-specific antigens with adjunct agents that either prolong antigen delivery (“depot effect”) and / or induce an appropriate immune stimulating effect, and thus enhance an immunological response against the cancer. One such adjunct agent is the cationic carbohydrate biopolymer chitosan, and another such agent is the semisynthetic biopolymer glycated chitosan (GC). Key advantageous properties of GC include improved water solubility and immune stimulation over base chitosans, sterile filterability, biocompatibility, and capability to serve as physiologically compatible carrier of other therapeutic agents (e.g. nanomaterials).

[0008] GC is prepared by reacting saccharide molecules with chitosan. Chitosan itself is produced from chitin, a structural component in many organisms, found for example in exoskeletons in arthropods, such as crustaceans and insects, and as cell walls in fungi. The biopolymer chitin is a linear homopolymer composed of A-acetylglucosamine units joined by P 1— >4 glycosidic bonds. Chitosan, which is partially deacetylated chitin, is the most studied member of this class of biopolymer-derived compounds. The presence of primary amino groups in chitosan facilitates a number of approaches for chemical modifications designed to achieve their solubilization and to impart special properties for specific applications. For example, in addition to improving its solubility, glycation of chitosan increases its immune- stimulating properties. Docket No.: IPW-00625

[0009] SUMMARY OF THE INVENTION

[0010] In certain aspects, the present disclosure relates to aqueous pharmaceutical formulations comprising chitosan or a modified chitosan, wherein the concentration of the chitosan or modified chitosan is about 0.1 mg / mL to about 2 mg / mL.

[0011] In some aspects, the present disclosure relates to glycated chitosans, wherein the glycated chitosan has a degree of glycation of about 0.1% to about 30%.

[0012] In certain aspects, the present disclosure relates to methods of enhancing an immune system response to an antigen in a patient in need thereof, comprising administering a pharmaceutical formulation or glycated chitosan disclosed herein.

[0013] BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1A is a bar graph showing the response of a murine model of colorectal cancer to exposure to varying concentrations of glycated chitosan with a 200 microliter injection volume.

[0015] Figure IB is a chart showing response of a murine model of colorectal cancer to exposure to varying concentrations of glycated chitosan with a 200 microliter injection volume.

[0016] Figure 2A is a bar graph showing the response of a murine model of colorectal cancer to exposure to varying concentrations of glycated chitosan with a 200 microliter injection volume.

[0017] Figure 2B is a chart showing response of a murine model of colorectal cancer to exposure to varying concentrations of glycated chitosan with a 100 microliter injection volume.

[0018] Figure 3 A is a bar graph depicting the concentration of CXCL10, as determined by ELISA, produced by cultured cells in response to stimulation with galactose, chitosan, or glycated chitosan with increasing levels of galactation (4% to 24% galactation). Peak CXCL10 production is induced at 12.5% galactation.

[0019] Figure 3B is a line graph depicting the concentration of CXCL10, as determined by ELISA, produced by cultured cells in response to stimulation with increasing concentrations of glycated chitosan (0.0625 pg / mL to 32 pg / mL) with increasing levels of galactation (4% to 24% galactation). Peak CXCL10 production is induced at 12.5% galactation.

[0020] Figure 3C is a line graph depicting the concentration of CXCL10, as determined by ELISA, produced by cultured cells in response to stimulation with increasing concentrations of glycated chitosan with varying molecular weights and 70%, 80%, or 95% deacetylation. Docket No.: IPW-00625

[0021] Peak CXCL10 production is induced at 70% deacetylation with both 220 kDa and 353 kDa formulations.

[0022] Figure 3D is a scatter plot with bar markers depicting the fold-change in AUC values over glycated chitosan with 4% galactation and 80% deacetylation as compared to chitosan and glycated chitosan with increasing levels of galactation. Left to right: 0% galactated, 4% galactated / 80% deacetylated, 12.5% galactated, and 24% galactated, as compared to galactose alone. Maximal fold-change in AUC over glycated chitosan with 4% galactation and 80% deacetylation is induced at 12.5% galactation.

[0023] Figure 3E is a scatter plot with bar markers. Each point in the scatter plot represents one replicate of an AUC experiment, representing the cumulative amount of CXCL10 obtained from THP-1 cell culture supernatants treated with 0.5, 1, 2, 4, 8, 15, and 32 pg / mL glycated chitosan. Left to right: 0% galactated (base chitosan), 4% galactated / 80% deacetylated, 12.5% galactated, and 24% galactated, as compared to galactose alone. Peak AUC values are achieved at 12.5% galactation.

[0024] Figure 4 is a bar graph depicting the concentration of CXCL10, as determined by ELISA, produced by cultured cells in response to stimulation with glycated chitosan with varying molecular weights and 70%, 80%, or 95% deacetylation. Peak CXCL10 production is induced at 70% deacetylation with both 220 kDa and 353 kDa formulations.

[0025] Figure 5 is a chart showing the combined response of a murine model of colorectal cancer CT26 and a murine model of melanoma B 16-F10 to exposure to varying concentrations of glycated chitosan with a 200 microliter injection volume. Data from 3 independent experiments were combined. Concentrations were grouped for comparison: 20-10mg / ml, 5- 1.25mg / ml, l-0.3125mg / ml, 0.156-0.1 mg / ml, ablation alone, untreated. Log-rank test was performed for statistical analysis.

[0026] Figure 6 contains 4 charts showing the response, represented by tumor volume, in a murine model of colorectal cancer CT26 to single radiation 10 days post tumor injection, with or without GC injected on day 11, followed by anti-PDl on D16, 21, 26 and 31 in select groups. At end of 90-day follow-up, the best performing group was 5 Gy + anti-PDl + 1 mg / ml GC with 4 animals without progressing tumors vs 2 in 5 Gy + anti-PD l , and 0 in 5 Gy + anti-PDl + lOmg / ml GC.

[0027] DETAILED DESCRIPTION OF THE INVENTION

[0028] The present disclosure provides, in various aspects, compositions and methods for treating cancer. Docket No.: IPW-00625

[0029] In some aspects, the present disclosure provides aqueous pharmaceutical formulations comprising chitosan or a modified chitosan, wherein the concentration of the chitosan or modified chitosan is about 0.1 mg / mL to about 2 mg / mL.

[0030] In some embodiments, the pharmaceutical formulations comprise a modified chitosan. In some embodiments, the modified chitosan is a glycated chitosan or a carboxyalkyl chitosan. In some embodiments, the concentration of the chitosan or modified chitosan is about 0.3 mg / mL to about 0.8 mg / mL. In certain embodiments, the concentration of the chitosan or modified chitosan is about 0.6 mg / mL to about 0.7 mg / mL.

[0031] In certain embodiments, the chitosan or modified chitosan has a molecular weight of about 50 kDa to about 300 kDa. In some embodiments, the chitosan or modified chitosan has a molecular weight of about 80 kDa to about 400 kDa. In some embodiments, the chitosan or modified chitosan has a molecular weight of about 80 kDa, about 115 kDa, about 175 kDa, about 195 kDa, about 220 kDa, about 280 kDa, or about 350 kDa.

[0032] In certain embodiments, the modified chitosan is glycated chitosan. In some embodiments, the glycated chitosan has a degree of deacetylation of about 60% to about 99%. In some embodiments, the glycated chitosan has a degree of deacetylation of about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99%. In some embodiments, the glycated chitosan has a degree of glycation of about 0.1% to about 30%. In certain embodiments, the degree of glycation is about 3% to about 6%. In certain embodiments, the degree of glycation is about 4% or about 5%. In certain embodiments, the degree of glycation is about 10% to about 30%. In some embodiments, the degree of glycation is about 12% to about 26%. In some embodiments, the degree of glycation is about 12% to about 16%. In certain embodiments, the degree of glycation is about 13% to about 15%. In some embodiments, the degree of glycation is about 10% to about 15%. In some preferred embodiments, the degree of glycation is about 11% to about 13%. In some particularly preferred embodiments, the degree of glycation is about 12.5%.

[0033] In some embodiments, the glycated chitosan is glycated with galactose.

[0034] In some embodiments, the pharmaceutical formulation is suitable for administration with a syringe comprising a hypodermic needle. In some embodiments, the hypodermic needle is an 18-gauge hypodermic needle, a 20-gauge hypodermic needle, or a 22-gauge hypodermic needle. In certain embodiments, the hypodermic needle is a 20-gauge hypodermic needle. For example, the pharmaceutical formulations described herein are of an appropriate viscosity for convenient parenteral administration through these needle gauges. In certain embodiments, the pharmaceutical formulations have a viscosity of about 5 centipoise (cP) to about 360 cP. In Docket No.: IPW-00625 certain embodiments, the pharmaceutical formulations have a viscosity of about 5 cP, about 25 cP, about 50 cP, about 75 cP, about 100 cP, about 125 cP, about 150 cP, about 175 cP, about 200 cP, about 225 cP, about 250 cP, about 275 cP, about 300 cP, about 325 cP, about 350 cP, or about 360 cP.

[0035] In some aspects, the present disclosure provides glycated chitosans, wherein the glycated chitosans have a degree of glycation of about 0.1% to about 30%. In certain embodiments, the degree of glycation is about 3% to about 6%. In some embodiments, the degree of glycation is about 4% or about 5%. In some embodiments, the degree of glycation is about 12% to about 26%. In certain embodiments, the degree of glycation is about 12% to about 16%. In some embodiments, the degree of glycation is about 10% to about 15%. In some preferred embodiments, the degree of glycation is about 11% to about 13%. In some particularly preferred embodiments, the degree of glycation is about 12.5%. In some embodiments, the glycated chitosan is glycated with galactose.

[0036] In certain aspects, the present disclosure provides methods of enhancing an immune system response to an antigen in a patient in need thereof, comprising administering a pharmaceutical formulation or glycated chitosan of any preceding claim.

[0037] In some embodiments, the patient has cancer.

[0038] In some aspects, the present disclosure provides methods of treating cancer, comprising administering a pharmaceutical formulation or glycated chitosan of the present disclosure.

[0039] In some embodiments, the patient has a cancer selected from colorectal cancer (CRC), nasopharyngeal cancer, lung cancer, non-small cell lung cancer (NSCLC), soft tissue sarcoma (STS), mammary sarcoma, melanoma, hepatocellular carcinoma (HCC), pancreatic cancer, renal cancer, breast cancer, prostate cancer, bladder cancer, head-and neck cancer, uveal melanoma, thyroid cancer, ovarian cancer, cervical cancer, cholangiocarcinoma, osteosarcoma, hepatobiliary carcinoma, brain cancer (e.g., glioblastoma, meningioma, astrocytoma, or neuroblastoma), cutaneous lymphoma, and esophageal cancer.

[0040] In some embodiments, the patient has a solid tumor, and further comprising administering the pharmaceutical formulation or glycated chitosan into a solid tumor.

[0041] In certain embodiments, the solid tumor has undergone ablation, adjuvant therapy (e.g., chemotherapy, hormone therapy, immunotherapy, or irradiation), or surgical removal or reduction.

[0042] In certain embodiments, the patient has an infectious disease. In some embodiments, the immune system response to an antigen is an interferon response. In some embodiments, the interferon response is a type I interferon response. Docket No.: IPW-00625

[0043] Chitosan is synthesized from chitin, a linear homopolymer composed of N- acetylglucosamine units joined by P-1-4 glycosidic bonds, by removing acetyl groups from chitin using various processes which include, but are not limited to, exposure to aqueous sodium hydroxide or potassium hydroxide.

[0044] Glycated chitosan refers to the products resulting from the reaction between the free amino groups of chitosan and the carbonyl groups of reducing monosaccharides and / or oligosaccharides. The products of this reaction (mainly a mixture of Schiff bases, i.e. the carbon atom of the carbonyl group double-bonded to the nitrogen atom of the amino group, and Amadori products, i.e. the carbon atom of said carbonyl group bonded to the nitrogen atom of said amino group by a single bond while an adjacent carbon atom is double-bonded to an oxygen atom) may be used as such or after stabilization by reduction with hydrides, such as sodium borohydride, or by exposure to hydrogen in the presence of suitable catalysts. These reactions are further described in US 5,747,475 and US 2018 / 312611, both of which are fully incorporated by reference herein.

[0045] As described above, glycated chitosan offers several biological advantages over unmodified chitosan. Among these advantages, glycated chitosan’s additional immune- stimulating capabilities, in addition to its improved solubility, allow for surprisingly low concentrations of glycated chitosan to elicit strong antineoplastic activity as compared to unmodified chitosan.

[0046] It has been surprisingly discovered herein that the antineoplastic activity of glycated chitosan is optimal at specific concentrations (for example, lower concentrations of glycated chitosan may have greater antineoplastic efficacy than higher concentrations of the same glycated chitosan). See, for example, Figs. 1, 2, and 3.

[0047] Glycated chitosan comprises repeat units selected from: ne Docket No.: IPW-00625

[0048] Pharmaceutical Compositions

[0049] The compositions and methods of the present invention may be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or the compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are for human administration, particularly for invasive routes of administration (i.e., routes, such as injection or implantation, that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipients can be chosen, for example, to affect delayed release of an agent or to selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form such as tablet, capsule (including sprinkle capsule and gelatin capsule), granule, lyophile for reconstitution, powder, solution, syrup, suppository, injection or the like. The composition can also be present in a transdermal delivery system, e.g., a skin patch. The composition can also be present in a soludon or suspension suitable for topical administration, such as a lotion, cream, or ointment.

[0050] A pharmaceutically acceptable carrier can contain physiologically acceptable agents that act, for example, to stabilize, increase solubility or to increase the absorption of a compound such as a compound of the invention. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a self-emulsifying drug delivery system or a self-micro-emulsifying drug delivery system. The pharmaceutical Docket No.: IPW-00625 composition (preparation) also can be a liposome or other polymer matrix, which can have incorporated therein, for example, a compound of the invention. Liposomes, for example, which comprise phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.

[0051] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0052] The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's soludon; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0053] A pharmaceutical composition (preparation) can be administered to a subject by any of a number of routes of administration including, for example, orally (for example, drenches as in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); via injection (for example, intradermally, intramuscularly, intraperitoneally, intravenously, intratumorally; percutaneously; or intraarterially); by absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (for example as a patch applied to the skin); intranasally (for example, as a nasal aerosol spray); and topically (for example, as a cream, ointment or spray applied to the skin). The compound may also be formulated for inhalation. In certain embodiments, a compound may be simply dissolved or suspended in Docket No.: IPW-00625 sterile water. Details of appropriate routes of administration and compositions suitable for same can be found in, for example, U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, as well as in patents cited therein.

[0054] The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 0.01 percent to about ninety- nine percent of active ingredient, preferably from about 0.01 percent to about 5 percent, most preferably from about 0.01 percent to about 0.2 percent, e.g. 0.1 percent.

[0055] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0056] Formulations of the invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), lyophile, powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. Compositions or compounds may also be administered as a bolus, electuary or paste.

[0057] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelati n capsules), tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium Docket No.: IPW-00625 carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as, modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

[0058] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surfaceactive or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0059] The tablets, and other solid dosage forms of the pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

[0060] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, microemulsions, solutions, suspensions, syrups and Docket No.: IPW-00625 elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

[0061] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0062] Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0063] Dosage forms for the topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required.

[0064] The ointments, pastes, creams and gels may contain, in addition to an active compound, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0065] Powders and sprays can contain, in addition to an active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0066] Transdermal patches have the added advantage of providing controlled delivery of a compound of the present invention to the body. Such dosage forms can be made by dissolving or dispersing the active compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel. Docket No.: IPW-00625

[0067] The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intratumoral, percutaneous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile aqueous or nonaqueous soludons, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents. In some embodiments, such pharmaceutically acceptable aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders may be hypotonic, isotonic, or hypertonic. A person of ordinary skill in the art will readily be able to determine the appropriate tonicity suitable for a given pharmaceutical composition.

[0068] Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0069] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

[0070] In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, Docket No.: IPW-00625 delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0071] Injectable depot forms are made by forming microencapsulated matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly (anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissue.

[0072] For use in the methods of this invention, active compounds can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.

[0073] Methods of introduction may also be provided by rechargeable or biodegradable devices. Various slow-release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form an implant for the sustained release of a compound at a particular target site.

[0074] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0075] The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound(s) being employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0076] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the pharmaceutical composition or compound 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. By “therapeutically effective amount” is meant the concentration of a compound that is sufficient to elicit the Docket No.: IPW-00625 desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the weight, sex, age, and medical history of the subject. Other factors which influence the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound of the invention. A larger total dose can be delivered by multiple administrations of the agent. Methods to determine efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13 ed., 1814-1882, herein incorporated by reference).

[0077] In general, a suitable daily dose of an active compound used in the compositions and methods of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0078] If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In certain embodiments of the present invention, the active compound may be administered two or three ti mes daily. In preferred embodiments, the active compound will be administered once daily.

[0079] The patient receiving this treatment is any animal in need, including primates, in particular humans; and other mammals such as equines, cattle, swine, sheep, cats, and dogs; poultry; and pets in general.

[0080] In certain embodiments, compounds of the invention may be used alone or conjointly administered with another type of therapeutic agent.

[0081] The present disclosure includes the use of pharmaceutically acceptable salts of compounds of the invention in the compositions and methods of the present invention. In certain embodiments, contemplated salts of the invention include, but are not limited to, 1- hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2- oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, 1- ascorbic acid, 1-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)- camphor- 10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, D-glucoheptonic acid, D-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric Docket No.: IPW-00625 acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, L-malic acid, malonic acid, mandelic acid, methanesulfonic acid, naphthalene- 1,5 -disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, proprionic acid, L-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid acid salts.

[0082] The pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent.

[0083] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0084] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0085] In some embodiments, the pharmaceutical formulations of the present disclosure may be formulated as aqueous solutions. In some such embodiments, the aqueous solutions may comprise a concentration of glycated chitosan ranging from about 0.01 mg / mL to about 100 mg / mL. In some preferred embodiments, the aqueous soludons may comprise a concentration of glycated chitosan of about 0.1 mg / mL to about 10 mg / mL, most preferably about 1 mg / mL. In some embodiments, the pharmaceutical formulations of the present disclosure may comprise various buffering agents such as those described herein. In some embodiments, the pharmaceutical formulations of the present disclosure comprise a suitable amount of an acid to provide a pharmaceutically acceptable pH. For example, the acid may be a carboxylic acid, such as acetic acid. In some embodiments, the glycated chitosan is present as the acetate salt. In some embodiments, the amount of the acid (e.g., the acetic acid) is sufficient to adjust the pH of the pharmaceutical formulation to between about 5.0 and about 6.0, most preferably between about 5.4 to about 5.6. In some embodiments, the pharmaceutical formulations of the Docket No.: IPW-00625 present disclosure have a pH of between about 5.0 and about 6.0, most preferably between about 5.4 to about 5.6.

[0086] In some embodiments, the pharmaceutical formulations of the present disclosure may be stored in sterile vials for individual administration. In some embodiments, the vials are plastic vials. In some embodiments, the volume of the vials suitable for storing the pharmaceutical formulations ranges from about 1 mL to about 10 mL. Most preferably the volume is about 5 mL.

[0087] In some embodiments, the pharmaceutical formulations of the present disclosure may be stored as pre-filled syringes. In some embodiments, the pre-filled syringes comprise a volume of the pharmaceutical formulation suitable for a single administration. In some embodiments, the volume of the pharmaceutical formulation in the pre-filled syringes ranges from about 0.5 mL to about 10 mL, most preferably from about 1 mL to about 4 mL. In some embodiments, the pre-filled syringes may comprise a luer-lock or luer-slip connection suitable for fitting a sterile hypodermic needle. In some embodiments, the pre-filled syringes may be fitted with a hypodermic needle of suitable gauge for administration of the pharmaceutical formulations, such as a 14-gauge, 16-gauge, 18-gauge, 20-gauge, 22-gauge, 24-gauge or 26- gauge hypodermic needle, most preferably an 18-gauge, 20-gauge, or 22-gauge hypodermic needle. The trained physician will readily be able to determine the suitability of a given gauge of hypodermic needle for passing the pharmaceutical formulations of the present disclosure into the body.

[0088] Definitions

[0089] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art. Generally, nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics and protein and nucleic acid chemistry, described herein, are those well-known and commonly used in the art.

[0090] The methods and techniques of the present disclosure are generally performed, unless otherwise indicated, according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout this specification. See, e.g. Motulsky, “Intuitive Biostatistics”, Oxford University Press, Inc. (1995); Lodish et al., “Molecular Cell Biology, 4th ed.”, W. H. Freeman & Co., New York (2000); Griffiths et al., “Introduction to Genetic Analysis, 7th ed.”, W. H. Freeman & Co., N.Y. Docket No.: IPW-00625

[0091] (1999); and Gilbert et al., “Developmental Biology, 6th ed.”, Sinauer Associates, Inc., Sunderland, MA (2000).

[0092] Chemistry terms used herein, unless otherwise defined herein, are used according to conventional usage in the art, as exemplified by “The McGraw-Hill Dictionary of Chemistry”, Editorial Staff of the McGraw-Hill Encyclopedia of Science & Technology, Ed., McGraw-Hill, New York, NY. (2003).

[0093] All of the above, and any other publications, patents and published patent applications referred to in this application are specifically incorporated by reference herein. In case of conflict, the present specification, including its specific definitions, will control.

[0094] The term “agent” is used herein to denote a chemical compound (such as an organic or inorganic compound, a mixture of chemical compounds), a biological macromolecule (such as a nucleic acid, an antibody, including parts thereof as well as humanized, chimeric and human antibodies and monoclonal antibodies, a protein or portion thereof, e.g., a peptide, a lipid, a carbohydrate polymer), or an extract made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues. Agents include, for example, agents whose structure is known, and those whose structure is not known.

[0095] A “patient,” “subject,” or “individual” are used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (including bovines, porcines, etc.), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats).

[0096] “Treating” a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread 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” can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0097] The term “preventing” is art-recognized, and when used in relation to a condition, such as a local recurrence (e.g., pain), a disease such as cancer, a syndrome complex such as heart failure or any other medical condition, is well understood in the art, and includes administration of a composition which reduces the frequency of, or delays the onset of, symptoms of a medical condition in a subject relative to a subject which does not receive the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment relative to an untreated Docket No.: IPW-00625 control population, and / or delaying the appearance of detectable cancerous growths in a treated population versus an untreated control population, e.g., by a statistically and / or clinically significant amount.

[0098] “Administering” or “administration of’ a substance, a compound or an agent to a subject can be carried out using one of a variety of methods known to those skilled in the art. For example, a compound or an agent can be administered, intravenously, arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, transdermally (by absorption, e.g., through a skin duct), and intratumorally. A compound or agent can also appropriately be introduced by rechargeable or biodegradable polymeric devices or other devices, e.g., patches and pumps, or formulations, which provide for the extended, slow or controlled release of the compound or agent. Administering can also be performed, for example, once, a plurality of times, and / or over one or more extended periods.

[0099] Appropriate methods of administering a substance, a compound or an agent to a subject will also depend, for example, on the age and / or the physical condition of the subject and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability and toxicity). In some embodiments, a compound or an agent is administered orally, e.g., to a subject by ingestion. In some embodiments, the orally administered compound or agent is in an extended release or slow release formulation, or administered using a device for such slow or extended release.

[0100] As used herein, the phrase “conjoint administration” refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered therapeutic agent is still effective in the body (e.g., the two agents are simultaneously effective in the patient, which may include synergistic effects of the two agents). For example, the different therapeutic compounds can be administered either in the same formulation or in separate formulations, either concomitantly or sequentially. Thus, an individual who receives such treatment can benefit from a combined effect of different therapeutic agents.

[0101] A “therapeutically effective amount” or a “therapeutically effective dose” of a drug or agent is an amount of a drug or an agent that, when administered to a subject will have the intended therapeutic effect. The full therapeutic effect does not necessarily occur by administration of one dose and may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount needed for a subject will depend upon, for example, the subject’s size, Docket No.: IPW-00625 health and age, and the nature and extent of the condition being treated, such as cancer or MDS. The skilled worker can readily determine the effective amount for a given situation by routine experimentation.

[0102] “Degree of glycation” as used herein refers to the number of glycated amine groups in a glycated chitosan divided by the total number of polymer subunits. When a degree of glycation is specified herein with respect to a polymer ensemble (for example a polymer mixture in a reaction or product mixture), that is a reference to percentage of glycated units in the ensemble with respect to the total number of units.

[0103] As used herein, a “number average” of a property of a polymer chain refers to the unweighted mean of that property across a polymer ensemble. Thus, for example, the “number average molecule weight” of a polymer ensemble may be expressed by the equation: where Ni is the number of molecules of molecular weight Mi. Weight average quantities may be determined by any suitable technique. For example, the number average molecular weight may be determined by gel permeation chromatography (also known as size exclusion chromatography) or viscometry. Other number averaged quantities may, for example, be derived from the number average molecular weight. For example, the number average of the number of repeat units in a polymer ensemble (also known as the number average degree of polymerization) may be calculated as the ratio of the number average molecular weight to the molecular weight of the repeat unit (appropriately averaged if necessary). The number average of the degree of polymerization of a polymer ensemble may also be measured directly, for example by end-group analysis.

[0104] As used herein, a “weight average” of a property of a polymer chain refers to the mean of that property across a polymer ensemble, weighted by the molecular weight of the polymer chains. Thus, for example, the “weight average molecular weight” of a polymer ensemble may be expressed by the equation: where Ni is the number of molecules of molecular weight Mi. Weight average quantities may be determined by any suitable technique. For example, the weight average molecular weight may be determined by light scattering, small angle neutron scattering, X-ray scattering, Docket No.: IPW-00625 or sedimentation velocity. Other weight averaged quantities may, for example, be derived from the weight average molecular weight. For example, the weight average of the number of repeat units in a polymer ensemble (also known as the weight average degree of polymerization) may be calculated as the ratio of the weight average molecular weight to the molecular weight of the repeat unit (appropriately averaged if necessary). The phrase “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filter, diluent, excipient, solvent or encapsulating material useful for formulati ng a drug for medicinal or therapeutic use.

[0105] The term “modified chitosan” includes non-covalent metal-chitosan complexes, alkylated chitosans, arylated chitosans, carboxyalkyl chitosans (e.g., carboxymethyl chitosan), hydroxyalkyl chitosans (e.g., hydroxypropyl chitosan, hydroxyethyl chitosan), aminoalkyl chitosans (such as secondary aminoalkyl chitosans and tertiary aminoalkyl chitosans), acylated chitosans, phosphorylated chitosan, thiolated chitosan, quaternary ammonium chitosans, guanidinylated chitosans, glycated chitosans (including mono- and oligosaccharides), chitosan poly (sulfonamides), chitosan poly(sulfonates) and chitosan-arylsuccinates.

[0106] EXAMPLES

[0107] The invention now being generally described, it will be more readily understood by reference to the following examples, which are included merely for purposes of illustration of certain aspects and embodiments of the present invention and are not intended to limit the invention.

[0108] Example 1. Exemplary Biological Evaluation of Compounds of the Disclosure

[0109] As shown in Figures 1A, IB, 2 A and 2B, in vivo efficacy testing of various different glycated chitosan concentrations was undertaken to evaluate the anti-cancer effect in a murine tumor model. CT26 murine colon cancer (purchased from ATCC) was implanted subcutaneously in the right and left flank of 6-8weeks old, female Balb / c mice. On day 7, animals were randomized and received micro wave thermal ablation of the right tumor followed by intratumoral injection of glycated chitosan (see Table 1 for the volumes and concentrations). The left tumor was left untreated as a surrogate of metastases. Growth of the tumors and survival were monitored over 84 days (Kaplan Meier Curve, log-rank (mantel-cox) test *p<0.05, ** p<0.01, ***p<0.001) . The bar graph in Figure la and 2a represents the percentage of animals that survived in each group up to the end of the monitoring period. Surprisingly, 0.625 mg / mL glycated chitosan was superior to other concentrations of glycated chitosan, even Docket No.: IPW-00625 in comparison to much higher concentrations. This outcome was observed regardless of whether 200 microliters (Figures 1A and IB) or 100 microliters (Figures 2 A and 2B) of glycated chitosan solution was injected. The experiment was repeated with the expanded range of concentrations at 200 microliters (Figure 5) and another tumor model B16-F10 mouse melanoma, combined data shows that overall survival of the animals was the highest in the ablation + IP-001 in the concentration range of l-0.3125mg / ml, in comparison to ablation alone (p=0.028), to 5-1.25mg / ml (p=0.03), or 20-10mg / ml (p=0.0007).

[0110] Table 1. Tabulated Survival Data for In Vivo Glycated Chitosan Efficacy Testing

[0111] For the in vitro assays depicted in Figures 3A-3E and 4, human monocyte cell line THP- 1 cells (purchased from Invivogen, Inc.) were thawed and cultured in complete media according to the manufacturer’s instructions. After fully recovered from thawing, THP-1 cells were harvested, washed and stimulated with chitosan or glycated chitosan of different concentrations (as specified in the graphs). For stimulation, THP-1 cells were placed in the cell culture incubator at 5% CO2, 37°C and 100% humidity for 48 hours. Supernatants were harvested and transferred to ELISA plates (purchased from Biolegend, Inc.) for assessment of CXCL10 according to the manufacturer’s instructions. Data from individual experiments was represented by bar graphs (Figures 3 A, 3D, 3E, 4) or line curves (Figures 3B, 3C). Data from multiple experiments (Figures 3D, 3E) was plotted as bar graphs where each data point Docket No.: IPW-00625 represented the AUC of CXCL10 from 0.5pg / ml-32pg / ml glycated chitosan in one experiment prior to analysis. Data are presented as mean + / - s.e.m. n=2 or more independent experiments. Statistical analysis was performed using Student’s t-test. *p<0.05, **p<0.005, ***p<0.0005, ****p<0.00005.

[0112] Another example of testing of different concentrations of GC in vivo was performed in conjunction of radiation and anti-PD- l . Animals were implanted with a mouse colorectal cancer cell line CT26 and treated with a single dose of radiation, on day 11, followed by anti- PD1 on D16, 21, 26 and 31 (Figure 6). At the end of a 90-day follow-up, the best performing group was 5 Gy + anti-PDl + Img / ml GC with 4 animals without progressing tumors vs 0 in 5Gy + anti-PDl + lOmg / ml GC.

[0113] INCORPORATION BY REFERENCE

[0114] All publications and patents mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

[0115] EQUIVALENTS

[0116] While specific embodiments of the subject invention have been discussed, the above specification is illustrative and not restrictive. Many variations of the invention will become apparent to those skilled in the art upon review of this specification and the claims below. The full scope of the invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations.

Claims

Docket No.: IPW-00625CLAIMSWe claim:

1. An aqueous pharmaceutical formulation comprising chitosan or a modified chitosan, wherein the concentration of the chitosan or modified chitosan is about 0.1 mg / mL to about 2 mg / mL.

2. The pharmaceutical formulation of claim 1, comprising a modified chitosan.

3. The pharmaceutical formulation of any one of the preceding claims, wherein the modified chitosan is a glycated chitosan or a carboxyalkyl chitosan.

4. The pharmaceutical formulation of any one of the preceding claims, wherein the concentration of the chitosan or modified chitosan is about 0.3 mg / mL to about 0.8 mg / mL.

5. The pharmaceutical formulation of any one of the preceding claims, wherein the concentration of the chitosan or modified chitosan is about 0.6 mg / mL to about 0.7 mg / mL.

6. The pharmaceutical formulation of any one of the preceding claims, wherein the chitosan or modified chitosan has a molecular weight of about 50 kDa to about 300 kDa.

7. The pharmaceutical formulation of any one of claims 1-5, wherein the chitosan or modified chitosan has a molecular weight of about 80 kDa to about 400 kDa.

8. The pharmaceutical formulation of claim 7, wherein the chitosan or modified chitosan has a molecular weight of about 80 kDa, about 115 kDa, about 175 kDa, about 195 kDa, about 220 kDa, about 280 kDa, or about 350 kDa.

9. The pharmaceutical formulation of any one of the preceding claims, wherein the modified chitosan is glycated chitosan.

10. The pharmaceutical formulation of claim 9, wherein the glycated chitosan has a degree of deacetylation of about 60% to about 99%.

11. The pharmaceutical formulation of claim 10, wherein the glycated chitosan has a degree of deacetylation of about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99%.Docket No.: IPW-0062512. The pharmaceutical formulation of claim 9, wherein the glycated chitosan has a degree of glycation of about 0.1% to about 30%.

13. The pharmaceutical formulation of claim 12, wherein the degree of glycation is about 3% to about 6%.

14. The pharmaceutical formulation of claim 13, wherein the degree of glycation is about 4% or about 5%.

15. The pharmaceutical formulation of claim 12, wherein the degree of glycation is about 10% to about 30%.

16. The pharmaceutical formulation of claim 15, wherein the degree of glycation is about 12% to about 26%.

17. The pharmaceutical formulation of claim 16, wherein the degree of glycation is about 12% to about 16%.

18. The pharmaceutical formulation of claim 17, wherein the degree of glycation is about 13% to about 15%.

19. The pharmaceutical formulation of claim 15, wherein the degree of glycation is about 10% to about 15%.

20. The pharmaceutical formulation of claim 19, wherein the degree of glycation is about 11% to about 13%.

21. The pharmaceutical formulation of any one of claims 12-20, wherein the glycated chitosan is glycated with galactose.

22. The pharmaceutical formulation of any one of claims 1-21, wherein the pharmaceutical formulation is suitable for administration with a syringe comprising a hypodermic needle.

23. The pharmaceutical formulation of claim 22, wherein the hypodermic needle is an 18- gauge hypodermic needle, a 20-gauge hypodermic needle, or a 22-gauge hypodermic needle.

24. The pharmaceutical formulation of claim 23, wherein the hypodermic needle is a 20- gauge hypodermic needle.Docket No.: IPW-0062525. A glycated chitosan, wherein the glycated chitosan has a degree of glycation of about 0.1% to about 30%.

26. The glycated chitosan of claim 25, wherein the degree of glycation is about 3% to about 6%.

27. The glycated chitosan of claim 26, wherein the degree of glycation is about 4% or about 5%.

28. The glycated chitosan of claim 25, wherein the degree of glycation is about 12% to about 26%.

29. The glycated chitosan of claim 28, wherein the degree of glycation is about 12% to about 16%30. The glycated chitosan of claim 25, wherein the degree of glycation is about 10% to about 15%.

31. The glycated chitosan of claim 30, wherein the degree of glycation is about 11% to about 13%.

32. The glycated chitosan of any one of claims 25-31, wherein the glycated chitosan is glycated with galactose.

33. A method of enhancing an immune system response to an antigen in a patient in need thereof, comprising administering a pharmaceutical formulation or glycated chitosan of any preceding claim.

34. The method of claim 33, wherein the patient has cancer.

35. A method of treating cancer, comprising administering a pharmaceutical formulation or glycated chitosan of any one of claims 1-34.

36. The method of any one of claims 33-35, wherein the patient has a cancer selected from colorectal cancer (CRC), nasopharyngeal cancer, lung cancer, non-small cell lung cancer (NSCLC), soft tissue sarcoma (STS), mammary sarcoma, melanoma, hepatocellular carcinoma (HCC), pancreatic cancer, renal cancer, breast cancer, prostate cancer, bladder cancer, head-and neck cancer, uveal melanoma, thyroid cancer, ovarian cancer, cervicalDocket No.: IPW-00625 cancer, cholangiocarcinoma, osteosarcoma, hepatobiliary carcinoma, brain cancer (e.g., glioblastoma, meningioma, astrocytoma, or neuroblastoma), cutaneous lymphoma, and esophageal cancer.

37. The method of any one of claims 33-35, wherein the patient has a solid tumor, and further comprising administering the pharmaceutical formulation or glycated chitosan into a solid tumor.

38. The method of claim 37, wherein the solid tumor has undergone ablation, adjuvant therapy (e.g., chemotherapy, hormone therapy, immunotherapy, or irradiation), or surgical removal or reduction.

39. The method of claim 33, wherein the padent has an infectious disease.

40. The method of any one of claims 33-39, wherein the immune system response to an antigen is an interferon response.

41. The method of claim 40, wherein the interferon response is a type I interferon response.

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