Description:FIELD OF THE INVENTION
[0001] The present disclosure generally relates to the field of pharmaceuticals. Specifically, it relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising PEG-coated chitosan hybrid nanoparticles loaded with one or more peroxisome proliferator-activated receptor gamma (PPAR-?) agonist; and a method for preparing the same.

BACKGROUND OF THE INVENTION
[0002] Background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.
[0003] Liver is the vital organ present in vertebrates whose function includes detoxification, protein synthesis, and production of biochemicals necessary for digestion. Liver has both parenchymal cells (hepatocytes) and non-parenchymal cells. About 80% of the liver volume is occupied by the parenchymal cells. Non – parenchymal cells occupy about 6.5% of the total liver volume.
[0004] Asialoglycoprotein receptors are found on the surface of parenchymal cells, which are responsible for the clearance of glycoproteins with acetylgalactosamine and disialylated galactose residues from the blood circulation by receptor-mediated endocytosis. Such receptors can be recognized by using some targeting ligands like galactosylated ligands, lactobionic ligands, asialofetuin ligand, soyabean derived sterylglucoside ligand.
[0005] Nanoparticles are polymeric particles made of natural or artificial polymers ranging in size between about 10 and 1000 nm (1 µm). Drugs may be bound in form of a solid solution or dispersion or be adsorbed to the surface or chemically attached. Nanoparticles provide massive advantages regarding drug targeting, delivery and release and, with their additional potential to combine diagnosis and therapy, emerge as one of the major tools in nanomedicine. The main goals are to improve their stability in the biological environment, to mediate the bio-distribution of active compounds, improve drug loading, targeting, transport, release, and interaction with biological barriers. The cytotoxicity of nanoparticles or their degradation products remains a major problem, and improvements in biocompatibility obviously is the main concern of future research.
[0006] The polymeric nanoparticles (PNPs) are prepared from biocompatible and biodegradable polymers in size between 10-1000 nm where the drug is dissolved, entrapped, encapsulated, or attached to a nanoparticle matrix. Depending upon the method of preparation nanoparticles, nanospheres or nanocapsules can be obtained. Nano-capsules are systems in which the drug is confined to a cavity surrounded by a unique polymer membrane, while nanospheres are matrix systems in which the drug is physically and uniformly dispersed.
[0007] The field of polymer nanoparticles (PNPs) is quickly expanding and playing an important role in a wide spectrum of areas ranging from electronics, photonics, conducting materials, sensors, medicine, biotechnology, pollution control, and environmental technology. PNPs are promising vehicles for drug delivery by easy manipulation to prepare carriers with the objective of delivering the drugs to specific targets, such an advantage improves drug safety. Polymer-based nanoparticles effectively carry drugs, proteins, and DNA to target cells and organs. Their nanometer size promotes effective permeation through cell membranes and stability in the bloodstream. Polymers are very convenient materials for the manufacture of countless and varied molecular designs that can be integrated into unique nanoparticles constructs with many potential medical applications.
[0008] Contemplated herein is a PEG-coated chitosan hybrid nanoparticles comprising therapeutic agents for hepatic targeting.

OBJECTS OF THE INVENTION
[0009] An object of the present invention is to provide a hepatic targeting pharmaceutical formulation comprising a therapeutic agent.
[0010] An object of the present invention is to provide a hepatic targeting pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist.
[0011] An object of the present invention is to provide a hepatic targeting pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation is nanoparticle-based formulation.
[0012] An object of the present invention is to provide a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation directly targets the liver.
[0013] An object of the present invention is to provide a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation comprises chitosan hybrid nanoparticles.
[0014] Another object of the present invention is to provide a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation comprises Lactobionic acid-conjugated chitosan nanoparticles.
[0015] Another object of the present invention is to provide a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation comprises Lactobionic acid-conjugated chitosan nanoparticles coated with PEG.
[0016] Another object of the present invention is to provide a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof, wherein the formulation directly targets the liver.
[0017] An object of the present invention is to provide a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof, wherein the formulation comprises chitosan hybrid nanoparticles.
[0018] Another object of the present invention is to provide a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof, wherein the formulation comprises Lactobionic acid-conjugated chitosan nanoparticles.
[0019] Another object of the present invention is to provide a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof, wherein the formulation comprises Lactobionic acid-conjugated chitosan nanoparticles coated with PEG.
[0020] Yet another object of the present invention is to provide a method for preparing a therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle that directly targets the liver.
[0021] Yet another object of the present invention is to provide a method for preparing a therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle comprises solvent evaporation method or solvent displacement method.
[0022] Yet another object of the present invention is to provide a method for polyethylene glycol (PEG) coating of therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle.
[0023] Yet another object of the present invention is to provide a method for preparing pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticle comprises solvent evaporation method or solvent displacement method.
[0024] Yet another object of the present invention is to provide a method for PEG coating of pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticles.
[0025] Yet another object of the present invention is to provide a method for PEG coating of pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticle, wherein the PEG coated nanoparticles directly targets liver.

SUMMARY OF THE INVENTION
[0026] This summary is provided to introduce a selection of concepts in a simplified form that are further described below in Detailed Description section. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter.
[0027] In an aspect, the present disclosure relates to a hepatic targeting pharmaceutical formulation comprising a therapeutic agent.
[0028] In an aspect, the present disclosure relates to a hepatic targeting pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist.
[0029] In an aspect, the present disclosure relates to a hepatic targeting pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation is nanoparticle-based formulation.
[0030] In an aspect, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation directly targets the liver.
[0031] In an aspect, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation comprises chitosan hybrid nanoparticles.
[0032] In another aspect, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation comprises Lactobionic acid-conjugated chitosan nanoparticles.
[0033] In another aspect, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation comprises Lactobionic acid-conjugated chitosan nanoparticles coated with PEG.
[0034] In another aspect, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof, wherein the formulation directly targets the liver.
[0035] In another aspect, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof, wherein the formulation comprises chitosan hybrid nanoparticles.
[0036] In another aspect, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof, wherein the formulation comprises Lactobionic acid-conjugated chitosan nanoparticles.
[0037] In another aspect, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof, wherein the formulation comprises Lactobionic acid-conjugated chitosan nanoparticles coated with PEG.
[0038] In yet another aspect, the present disclosure relates to a method for preparing a therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle that directly targets the liver.
[0039] In yet another aspect, the present disclosure relates to a method for preparing a therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle comprises solvent evaporation method or solvent displacement method.
[0040] In yet another aspect, the present disclosure relates to a method for polyethylene glycol (PEG) coating of therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle.
[0041] In yet another aspect, the present disclosure relates to a method for polyethylene PEG coating of therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle that directly targets the liver.
[0042] In yet another aspect, the present disclosure relates to a method for preparing pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticle comprises solvent evaporation method or solvent displacement method.
[0043] In yet another aspect, the present disclosure relates to a method for PEG coating of pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticle.
[0044] In yet another aspect, the present disclosure relates to a method for PEG coating of pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticle, wherein the PEG-coated nanoparticles directly targets the liver of a subject.
[0045] Other aspects of the invention will be set forth in the description which follows, and in part will be apparent from the description, or may be learnt by the practice of the invention.

DETAILED DESCRIPTION
[0046] The following is a detailed description of embodiments of the disclosure. The embodiments are in such detail as to clearly communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure as defined by the appended claims.
[0047] All publications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply.
[0048] Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
[0049] In some embodiments, numbers have been used for quantifying weight percentages, angles, and so forth, to describe and claim certain embodiments of the invention and are to be understood as being modified in some instances by the term “about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. The numerical values presented in some embodiments of the invention may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.
[0050] Various terms as used herein are shown below. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
[0051] As used in the description herein and throughout the claims that follow, the meaning of “a,” “an,” and “the” includes plural reference unless the context clearly dictates otherwise. Also, as used in the description herein, the meaning of “in” includes “in” and “on” unless the context clearly dictates otherwise.
[0052] Unless the context requires otherwise, throughout the specification which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense that is as “including, but not limited to.”
[0053] The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein.
[0054] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. “such as”) provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.
[0055] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group can be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is herein deemed to contain the group as modified.
[0056] The description that follows, and the embodiments described therein, is provided by way of illustration of an example, or examples, of particular embodiments of the principles and aspects of the present disclosure. These examples are provided for the purposes of explanation, and not of limitation, of those principles and of the disclosure.
[0057] The headings and abstract of the invention provided herein are for convenience only and do not interpret the scope or meaning of the embodiments.
[0058] The following discussion provides many example embodiments of the inventive subject matter. Although each embodiment represents a single combination of inventive elements, the inventive subject matter is considered to include all possible combinations of the disclosed elements. Thus if one embodiment comprises elements A, B, and C, and a second embodiment comprises elements B and D, then the inventive subject matter is also considered to include other remaining combinations of A, B, C, or D, even if not explicitly disclosed.
[0059] As described herein, the term “therapeutic agent” has the meaning known in the state of the art. The term denotes a pharmaceutical drug or compound that produces a desired biological activity in the body.
[0060] As described herein, the term “nanoparticle” has the meaning present in the state of the art. The term denotes a nanometer sized particulates in which the pharmaceutical actives are present.
[0061] While a particular form of the invention has been illustrated and described, it will be apparent that various modifications can be made without departing from the spirit and scope of the invention.
[0062] The present disclosure generally relates to pharmaceutical formulations and processes of preparing the same for treating various medical conditions.
[0063] In an embodiment, the present disclosure relates to a hepatic targeting pharmaceutical formulation comprising a therapeutic agent or a pharmaceutically acceptable salt thereof.
[0064] In an embodiment, the present disclosure relates to a hepatic targeting pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist.
[0065] In an embodiment, the present disclosure relates to a hepatic targeting pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation is nanoparticle-based formulation.
[0066] In an embodiment, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising a therapeutic agent or a pharmaceutically acceptable salt thereof, wherein the nanoparticles directly target the liver.
[0067] In an embodiment, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation directly targets the liver.
[0068] In an embodiment, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation comprises modified chitosan nanoparticles.
[0069] In an embodiment of the present disclosure, the modified chitosan comprises conjugated chitosan.
[0070] In an embodiment of the present disclosure, the conjugated chitosan comprises a liver targeting moiety conjugated chitosan.
[0071] In an embodiment of the present disclosure, the hepatic targeting moiety is selected from galactosylated ligands, lactobionic acid ligands, asialofetuin ligand, soyabean derived sterylglucoside ligand, or combinations thereof. More preferably Lactobionic acid ligands.
[0072] In preferred embodiment of the present disclosure, the conjugated chitosan comprises Lactobionic acid-conjugated chitosan.
[0073] In preferred embodiment of the present disclosure, the hepatic targeting nanoparticle-based pharmaceutical formulation comprises Lactobionic acid-conjugated chitosan nanoparticles.
[0074] In another embodiment of the present disclosure, the Lactobionic acid-conjugated chitosan nanoparticle is prepared by mixing Lactobionic acid and chitosan in presence of EDC (1-Ethyl-3-[3-dimethylami-nopropyl]-carbodiimide hydrochloride) and NHS (N-hydroxy succinimide) at a pH of 6, followed by magnetic stirring for 24 h at 25 deg C.
[0075] In another embodiment of the present disclosure, the Lactobionic acid-conjugated chitosan nanoparticle is loaded with a therapeutic agent, wherein the nanoparticles directly target the liver of a subject, wherein the subject is human or animal.
[0076] In another embodiment of the present disclosure, the Lactobionic acid-conjugated chitosan nanoparticle is loaded with peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof.
[0077] In another embodiment of the present disclosure, a method for preparing Lactobionic acid-conjugated chitosan nanoparticles loaded with peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof by solvent evaporation method or solvent displacement method.
[0078] In another embodiment of the present disclosure, a method for preparing a therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle comprises the steps of:
a. mixing Lactobionic acid-conjugated chitosan nanoparticles and a therapeutic agent to prepare hydrophilic phase;
b. preparing an organic phase comprising dichloromethane as an organic solvent and SPAN 20 (sorbitan monolaurate);
c. adding hydrophilic phase to the organic phase, dropwise, under continuous stirring at 3000 rpm for overnight; and
d. homogenizing the resulting solution comprising therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticles.
[0079] For the purpose of this invention SPAN 20 is Sorbitan monolaurate which is a mixture of esters formed from the fatty acid lauric acid and polyols derived from sorbitol, including sorbitan and isosorbide.
[0080] In another embodiment of the present disclosure, instead of SPAN 20 (sorbitan monolaurate) other surfactant such as medium chain triglycerides, polysorbate 80, colloidal anhydrous silica, and xanthan gum can be used.
[0081] In another embodiment of the present disclosure, the therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle is coated with polyethylene glycolate (PEG) by PEGylation.
[0082] In another embodiment of the present disclosure, a method for PEGylating a therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle comprises the steps of:
a. dissolving of PEG with water for injection;
b. mixing the therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle and the solution from step a, followed by stirring overnight;
c. adding hydrophilic phase to the organic phase, dropwise, under continuous stirring at 3000 rpm for overnight; and
d. ultracentrifugation to remove un-reacted PEG in supernatant solution; and
e. Harvesting the precipitate comprising therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticles coated with PEG.
[0083] In an embodiment of the present disclosure, the therapeutic agent is selected from but not limited to dapagliflozin propanediol, dapagliflozin, liraglutide, GR-MD-02, semaglutide, cenicriviroc, F-351, peg-ilodecakin, ipragliflozin, ursodeoxycholic acid, colesevelam, pioglitazone, VK-2809, emricasan, linagliptin, elafibranor, DS-102, Px-102, Px-103, GS-4997, simtuzumab, DUR-928, mercaptamine, olesoxime, cobiprostone, bertilimumab, MDV-4463, irbesartan, GS-9674, BOT-191, MGL-3196, BMS-986171, PEG-FGF21, LJN-452, CF-102, KD-025, volixibat, volixibat potassium ethanolate hydrate, aramchol, tipelukast, NGM-313, FG-3019, CAT-2003, NGM-282, TRX-318, IONIS-DGAT2Rx, IMM-124-E, RG-125, norursodeoxycholic acid, KBP-042, leucine, metformin, sildenafil, A-4250, GKT-831, BB-3, saroglitazar, BG-00011, alipogene tiparvovec, MB-12066, betaine anhydrous, ARI-3037MO, HepaStem, PXS-4728A, CIGB-500, oltipraz, omega-3 carboxylic acids, dapagliflozin, remogliflozin etabonate, remogliflozin, LC-280126, JKB-121, DWP-10292, VBY-376, VBY-825, icosapent ethyl ester, Fuzheng Huayu capsule, interferon gamma, acetylsalicylic acid, hydrochlorothiazide, enalapril, atorvastatin, NC-101, TCM-606F, obeticholic acid, INT-767, GNF-5120, cryptoc hinone-D, fexaramine, caprylic triglyceride, evogliptin, GM-CT-01, high dose vitamin E (>400 iU/d) or a pharmaceutically acceptable salt thereof.. Most preferably pioglitazone.
[0084] In preferred embodiment of the present disclosure, the Lactobionic acid-conjugated chitosan nanoparticle is loaded with pioglitazone.
[0085] In preferred embodiment of the present disclosure, a method for preparing a pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticle comprises the steps of:
a. mixing Lactobionic acid-conjugated chitosan nanoparticles and pioglitazone to prepare hydrophilic phase;
b. preparing an organic phase comprising dichloromethane as an organic solvent and SPAN 20;
c. adding hydrophilic phase to the organic phase, dropwise, under continuous stirring at 3000 rpm for overnight; and
d. homogenizing the resulting solution comprising the pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticles.
[0086] In a preferred embodiment of the present disclosure, a pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticle is coated with polyethylene glycolate (PEG) by PEGylation.
[0087] In a preferred embodiment of the present disclosure, a method for PEGylating a pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticle comprises the steps of:
a. dissolving of PEG with water for injection;
b. mixing the pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticle and the solution from step a, followed by stirring overnight;
c. adding hydrophilic phase to the organic phase, dropwise, under continuous stirring at 3000 rpm for overnight; and
d. ultracentrifugation to remove un-reacted PEG in supernatant solution; and
e. removing precipitate comprising pioglitazone-loaded Lactobionic acid-conjugated chitosan nanoparticles coated with PEG.
[0088] In an embodiment, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone in weight of at least 50 mg, at least 100 mg, at least 150 mg, at least 200 mg, at least 250 mg, at least 300 mg, at least 350 mg, at least 400 mg, at least 450 mg, at least 500 mg. More preferably at least 300-500 mg.
[0089] In an embodiment, the present disclosure relates to a hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone in a weight of at least 1-2 mg/kg/day, 3-4 mg/kg/day, at least 5-6 mg/kg/day, at least 7-8 mg/kg/day, at least 9-10 mg/kg/day, at least 1-5 mg/kg/day, at least 5-10 mg/kg/day, at least 10-15 mg/kg/day, at least 20-25 mg/kg/day, at least 25-30 mg/kg/day, at least 30-35 mg/kg/day, at least 35-40 mg/kg/day, at least 40-45 mg/kg/day, at least 45-50 mg/kg/day of the subject. More preferably at least 5-10 mg/kg/day of the subject.
[0090] In an embodiment of the present disclosure, the hepatic targeting nanoparticle-based pharmaceutical formulation is used to treat one or more disorders, and wherein the disorder is selected from but not limited to the group consisting of non-alcoholic fatty liver disease, drug-induced liver injury, autoimmune liver disease, liver cancer, viral hepatitis, alcoholic liver and the like, liver cirrhosis, obesity-related metabolic syndrome and cardiovascular and cerebrovascular disease, dyslipidemia, familial hypercholesterolemia, obesity, metabolic syndrome, diabetes selected from type 1 or type 2 diabetes, and combinations thereof. Most preferably non-alcoholic fatty liver disease.
[0091] In one embodiment, nanoparticles comprise particle size is in the range 10 nm to 5000 nm, such as 10 to 500 nm, particularly 100 or 300 nm, for example, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% such as at least 99% of the particles are the relevant size or within said range. Thus in one embodiment of the invention at least 95%, at least 98% or at least 99% of the particles of the pharmaceutical composition have a diameter of between 10 and 500 nm. In another embodiment at least 95%, at least 98% or at least 99% of the particles of the pharmaceutical composition have a diameter of between 100 and 300 nm.
[0092] In one embodiment, the hepatic targeting nanoparticle-based pharmaceutical formulation does not contain particles higher than 1000 nm in diameter.
[0093] In an embodiment of the present disclosure, the hepatic targeting nanoparticle-based pharmaceutical formulation may further comprise additional active ingredient(s) selected from one or more of group consisting of: ace-inhibitors, anti-Alzheimer's agents, antianginal drugs, anti-arrhythmias, anti-asthmatics, anti-cholesterolemics, analgesics, anesthetics, anti-convulsants, anti-depressants, anti-diabetic agents, anti-diarrhea preparations, antidotes, anti-emetics, anti-histamines, anti-hypertensive drugs, anti-inflammatory agents, anti-lipid agents, anti-manics, anti-migraines, anti-nauseants, anti-stroke agents, anti-thyroid preparations, anti-tumor drugs, anti-viral agents, acne drugs, alkaloids, amino acid preparations, anti-tussives, anti-uricemic drugs, anti-viral drugs, anabolic preparations, systemic and non-systemic anti-infective agents, anti-neoplastics, anti-parkinsonian agents, anti-rheumatic agents, anxiolytics, anti-psychotics, appetite stimulants, biological response modifiers, blood modifiers, bone metabolism regulators, bronchodilators, cardiovascular agents, central nervous system stimulates, cholinesterase inhibitors, contraceptives, decongestants, dietary supplements, dopamine receptor agonists, endometriosis management agents, enzymes, erectile dysfunction agents, fertility agents, gastrointestinal agents, H2-antagonists, homeopathic remedies, hormones, hypercalcemia and hypocalcemia management agents, immunomodulators, immunosuppressives, migraine preparations, motion sickness treatments, muscle relaxants, non-steroidal anti-inflammatories (NSAID's), obesity management agents, osteoporosis preparations, oxytocics, parasympatholytics, parasympathomimetics, prostaglandins, psychotherapeutic agents, respiratory agents, sedatives, serotonin 5-HT3 receptor antagonists, smoking cessation aids, sympatholytics, tremor preparations, urinary tract agents, vasodilators, laxatives, antacids, ion exchange resins, anti-pyretics, appetite suppressants, expectorants, anti-anxiety agents, anti-ulcer agents, anti-inflammatory substances, coronary dilators, cerebral dilators, peripheral vasodilators, psycho-tropics, stimulants, anti-hypertensive drugs, vasoconstrictors, migraine treatments, antibiotics, tranquilizers, anti-psychotics, anti-tumor drugs, anti-coagulants, anti-thrombotic drugs, hypnotics, anti-emetics, anti-nauseants, anti-convulsants, neuromuscular drugs, hyper- and hypo-glycemic agents, thyroid and anti-thyroid preparations, diuretics, anti-spasmodics, anti-obesity drugs, erythropoietic drugs, anti-asthmatics, cough suppressants, mucolytics, DNA and genetic modifying drugs, and combinations thereof.
[0094] In a preferred embodiment, the frequency of administration of the hepatic targeting nanoparticle-based pharmaceutical formulation may vary, depending upon personal or medical needs, but generally ranges from about once per day to about four times daily.
[0095] In an embodiment of the present disclosure, the hepatic targeting nanoparticle-based pharmaceutical formulation further comprise at least one pharmaceutically acceptable excipient.
[0096] In an embodiment of the present disclosure, the hepatic targeting nanoparticle-based pharmaceutically acceptable excipients include but are not limited to, binders, diluents, thickeners, flow agents, absorbents, disintegrants, superdisintegrants, preservatives, emulsifiers, taste makers, stabilizers, sugars, anti-foaming agents, anti-caking agents, lubricants, coloring agents, coating materials, and combinations thereof. The excipients may be selected from those well known in the art.
[0097] In an embodiment of the present disclosure, the hepatic targeting nanoparticle-based pharmaceutical formulation may further comprise sugars selected from the group comprising of sucrose, glucose, dextrose, lactose, fructose, maltose, galactose, maltodextrin and the like.
[0098] In an embodiment of the present disclosure, the hepatic targeting nanoparticle-based pharmaceutical formulation can be formulated in dosage forms, including, liquids, suspensions, semi-solids, solutions, syrups, gels, emulsions, and the like.
[0099] In another embodiment, the present disclosure relates to a method of treatment, amelioration, or prevention of one or more liver diseases by administering the pharmaceutically effective amount of the hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof.
[00100] In a preferred embodiment, the present disclosure relates to a method of treatment, amelioration, or prevention of non-alcoholic fatty liver disease by administering the pharmaceutically effective amount of the hepatic targeting nanoparticle-based pharmaceutical formulation comprising pioglitazone or pharmaceutically active salts thereof.
[00101] In a preferred embodiment, the present disclosure relates to a method of treatment, amelioration, or prevention of non-alcoholic fatty liver disease by administering the pharmaceutically effective amount of the Lactobionic acid-conjugated chitosan nanoparticles coated with PEG loaded with peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof.
[00102] In a preferred embodiment, the present disclosure relates to a method of treatment, amelioration, or prevention of non-alcoholic fatty liver disease by administering the pharmaceutically effective amount of the Lactobionic acid-conjugated chitosan nanoparticles coated with PEG loaded with pioglitazone or pharmaceutically active salts thereof.
[00103] While the foregoing describes various embodiments of the disclosure, other and further embodiments of the disclosure may be devised without departing from the basic scope thereof. The scope of the invention is determined by the claims that follow. The invention is not limited to the described embodiments, versions, or examples, which are included to enable a person having ordinary skill in the art to make and use the invention when combined with information and knowledge available to the person having ordinary skill in the art.
[00104] In an embodiment, the pharmaceutical composition of the present invention maybe used in any manner known to a person skilled in the art.
EXAMPLES
[00105] The present disclosure is further explained in the form of the following examples. However, it is to be understood that the foregoing examples are merely illustrative and are not to be taken as limitations upon the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications may be made without departing from the scope of the invention.
[00106] Example 1: Preparation of pioglitazone loaded Nanoparticles solvent evaporation method (solvent displacement method).
50 mg of drug added to about 70 ml. of the prepared Lactobionic acid-conjugated chitosan solution.
Organic phase prepared in another separate beaker using 50 ml of dichloromethane as an organic solvent and add 2 ml SPAN 20.
Hydrophilic phase added dropwise to the organic phase with continuous stirring at 3000 rpm, and continued the stirring overnight.
The solution is then homogenized to prepare the pioglitazone-loaded nanoparticles.
[00107] Example 2: Coating of Nanoparticles with Polyethylene Glycol (PEG):
50 mg of PEG dissolved in water for injection (isotonicity and pH maintained well), shaken well. The prepared 25 mg of nanoparticles from Example 1 added to the prepared solution and mixed well by stirring overnight using a mechanical stirrer.
The excess PEG removed by centrifugation in the supernatant solution (unreacted PEG).
PEG nanoparticles were recovered from the precipitate.

ADVANTAGES OF THE PRESENT INVENTION
[00108] The present disclosure provides a hepatic targeting pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof that satisfies the existing needs, as well as others, and generally overcomes the deficiencies found in the prior art.
[00109] The present disclosure provides a hepatic targeting pharmaceutical formulation that directly targets the liver so that maximum drug reaches the target site.
[00110] The present disclosure provides a hepatic targeting system in a nanosize range that can be helpful to increase the stability of any volatile pharmaceutical agents, easily and cheaply fabricated in large quantities by a multitude of methods.
[00111] The present disclosure provides a hepatic targeting system comprising biodegradable materials for nanoparticle preparation, allows sustained drug release at the targeted site after injection over a period of days or even weeks.
[00112] The present disclosure provides a hepatic targeting system comprising polymeric nanoparticles that have made them ideal candidates for cancer therapy, delivery of vaccines, contraceptives, and delivery of targeted antibiotics.
[00113] The present disclosure provides a hepatic targeting system comprising polymeric nanoparticles that have made them ideal candidates for tissue engineering.
[00114] The present disclosure provides a hepatic targeting system comprising polymeric nanoparticles that can easily penetrate into even small capillaries and are taken up within cells, allowing an efficient drug accumulation at the targeted sites in the body.
[00115] The present disclosure provides a hepatic targeting system that reduces dose-related toxicity and increases efficacy.
[00116] The present disclosure provides a hepatic targeting system comprising Lactobionic acid that is easily recognized by the liver receptors known as asialoglycoprotein receptors, which cover about 80% of liver cell’s receptors, and hence it is very helpful in achieving maximum liver targeting.
[00117] The present disclosure provides a hepatic targeting system comprising PEG coating that avoids the opsonization process in the body and hence long circulation time can be achieved. Also, PEGylation is very helpful in avoiding the re-agglomeration of prepared nanoparticles.
, Claims:1. A hepatic targeting nanoparticle-based pharmaceutical formulation comprising peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof, wherein the formulation comprises modified chitosan nanoparticles.
2. The formulation as claimed in claim 1, wherein the modified chitosan nanoparticles comprise conjugated chitosan.
3. The formulation as claimed in claim 2, wherein the conjugated chitosan comprises a hepatic targeting moiety conjugated chitosan.
4. The formulation as claimed in claim 3, wherein the hepatic targeting moiety is selected from a group comprising galactosylated ligands, lactobionic acid ligands, asialofetuin ligand, soyabean derived sterylglucoside ligand, or combinations thereof.
5. The formulation as claimed in claim 2, wherein the conjugated chitosan comprises Lactobionic acid-conjugated chitosan.
6. The formulation as claimed in claim 5, wherein the peroxisome proliferator-activated receptor gamma (PPAR-?) agonist or pharmaceutically active salts thereof is the Lactobionic acid-conjugated chitosan loaded with a therapeutic agent.
7. The formulation as claimed in claim 6, wherein the therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle is coated with polyethylene glycolate (PEG) by PEGylation.
8. A method for preparing a therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle, comprises the steps of:
a) mixing Lactobionic acid-conjugated chitosan nanoparticles and a therapeutic agent to prepare hydrophilic phase;
b) preparing an organic phase comprising dichloromethane as an organic solvent and SPAN 20;
c) adding hydrophilic phase to the organic phase, dropwise, under continuous stirring at 3000 rpm for overnight; and
d) homogenizing the resulting solution comprising therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticles.
9. A process for preparing Lactobionic acid-conjugated chitosan nanoparticle, comprises the steps of
a) mixing Lactobionic acid and chitosan in presence of EDC (1-Ethyl-3-[3-dimethylami-nopropyl]-carbodiimide hydrochloride) and NHS (N-hydroxy succinimide);
b) maintaining pH of the mixture of step a) at 6; and
c) stirring the mixture of step a) for 24 hours at 25 deg C.
10. A method for PEGylating a therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle comprises the steps of:
a. dissolving of PEG with water for injection;
b. mixing the therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticle and the solution from step a, followed by stirring overnight;
c. adding hydrophilic phase to the organic phase, dropwise, under continuous stirring at 3000 rpm for overnight; and
d. ultracentrifugation to remove un-reacted PEG in supernatant solution; and harvesting the precipitate comprising therapeutic agent-loaded Lactobionic acid-conjugated chitosan nanoparticles coated with PEG.