DESC:FIELD OF INVENTION:
The present invention relates to pharmaceutical compositions comprising polymeric nanoparticles of anticancer drugs, a process for preparing such pharmaceutical compositions and their use in the treatment of liver cancer, specifically hepatocellular carcinoma.

BACKGROUND AND PRIOR ART:
Liver cancer is one of the most widespread forms of cancer and the major reason for tumor related mortality. Globally, more than 6,00,000 cases of death per year are associated with hepatic cancer. Although, liver cancer is observed more commonly in the developing countries, its instances are increasing at an alarming rate throughout the world.

Hepatocellular carcinoma comprises of around 95% of the primary liver cancers followed by cholangiocarcinoma (3.4%) and other cancers (1%). Hence, most of the antineoplastic treatments in patients are directed towards hepatocellular carcinoma.

Current treatments for hepatocellular carcinoma include surgical resection, chemotherapy, liver transplant, chemoembolization, trans-arterial embolization, radiation therapy, ethanol injection and cryoablation.

Liver resection is the best suitable treatment for non-cirrhotic liver cancer subjects. The chances of resection and post-operative survival are better in non-cirrhotic patients than those suffering from cirrhosis. However, this therapy holds limitations like suitability for small tumors. Liver transplantation proves to be a boon for patients with small tumor size (<5 cm), but suffers from restricted amount of donors and delay due to the donor non-availability.

Chemotherapy although widely used to treat tumors, results in more side effects and toxicity due to the destruction of normal, non-malignant cells. Drug delivery to the tumors occurs through endothelial junctions, fenestrations and vesicular organelles. Factors like P-glycoprotein mediated drug efflux and changes in enzyme activity add to failure via chemotherapy. Pharmacokinetics is an important parameter to be considered during chemotherapy. Ideally, cancerous cells should be exposed to suitable concentrations of drug over an extended time period. Current chemotherapy treatments are given over a short period of time with time intervals of 2-3 weeks. This results in speedy growth of tumors and thus diminishes the therapeutic effects of drugs with untoward side effects. Also, it was seen that prolonged drug exposure to tumors produced better results rather than short term exposure to elevated drug concentrations. Currently, the chemotherapy of hepatocellular carcinoma is performed by use of anthracycline antibiotics like doxorubicin, mitomycin, alkylating agent like cisplatin and other drugs like floxuridine, bevacizumab and sorafenib tosylate.

Percutaneous ethanol injection is an excellent option for small tumors; however it cannot be used in case of patients with ascites. Radiofrequency ablation has a curative effect on liver cancer because of lesser sessions required for therapy, but possesses more complications than percutaneous ethanol injection. Therapeutic effects with other treatments prove to be inadequate due to metastasis of tumors, lack of suitable embolizing materials, non-specificity to tumor cells and greater number of complications.

In spite of various therapies available for hepatocellular carcinoma, none have sufficed in complete and side effect free treatment of hepatic cancer. In this respect, many other alternative treatments have been explored for treatment of liver cancer such as nanoparticles.

Nanoparticles are minute, solid particulates in the size range of 10-1000 nm where, an anticancer agent is either entrapped in the core, adsorbed on the surface or both. Nanospheres or nanocapsules could be prepared by varying the methodology for synthesis of these colloidal carriers. The aim of any cancer treatment is to ensure optimum drug delivery to the desired cancerous site. The potential of using nanoparticles for delivery of anticancer drugs is massive and can be applied to various areas of medicine.

Gold Nanoparticles Conjugated with Cisplatin/Doxorubicin/Capecitabine Lower the Chemoresistance of Hepatocellular Carcinoma-Derived Cancer Cells, Tomuleasa et al, J Gastrointestin Liver Dis, June 2012 Vol. 21 No 2, 187-196.
Nanotechnology in oncology: Characterization and in vitro release kinetics of cisplatin-loaded albumin nanoparticles: Implications in anticancer drug delivery, Indian J Pharmacol. 2011 Jul-Aug; 43(4): 409–413.

Protein Nanoparticles as Drug Delivery Carriers for Cancer Therapy, BioMed Research International, Warangkana Lohcharoenkal et al, Volume 2014 (2014).

Although the prior art discloses various therapies , techniques and formulations for the treatment of hepatocellular carcinoma, each therapy , technique and formulation has a setback due to its side effects, administration type and regime or formulation feasibility and stability. Nanoparticles disclosed in prior art though efficacious would pose to have cost related implications. Hence there exists a need for development of nanoparticulate pharmaceutical compositions comprising anticancer drugs which can alleviate the above problems as well as target such anticancer drugs at the specific site of action.

OBJECT OF THE INVENTION:
An object of the present invention is to provide pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs.

Another object of the present invention is to provide a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs optionally with one or more pharmaceutically acceptable excipients.

Another object of the present invention is to provide a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs exhibiting prolonged release thereby leading to reduced frequency of dosing.

Another object of the present invention is to provide a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs having improved surface area and solubility.

Another object of the present invention is to provide a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs having a reduced dose.
Another object of the present invention is to provide a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs with minimized irritation potential at the site of action thus leading to increased patient compliance.

Another object of the present invention is to provide a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs to deliver the said anticancer drugs to the target site of action as well as attempt to decrease or avoid the side effects.

Another object of the present invention is to provide a process for preparing a pharmaceutical composition comprising homogenous polymeric nanoparticles of anticancer drugs optionally with one or more pharmaceutically acceptable excipients.
Another object of the present invention is to provide the use in the treatment of hepatocellular carcinoma by administering a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs.

Another object of the present invention is to provide the use of a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs and optionally one or more pharmaceutically acceptable excipients in the manufacture of a medicament for treatment of hepatocellular carcinoma.

Another object of the present invention is to provide a method of treating hepatocellular carcinoma by administering a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs.

SUMMARY OF THE INVENTION:
According to an aspect of the present invention, there is provided a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs and optionally one or more pharmaceutically acceptable excipients.

According to another aspect of the invention, there is provided a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs to deliver said anticancer drugs to the target site of action as well as attempt to decrease or avoid the side effects.
According to another aspect of the invention, there is provided a process for the preparation of a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs and optionally one or more pharmaceutically acceptable excipients.

According to an aspect of the present invention, there is provided a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs and optionally one or more pharmaceutically acceptable excipients for use in the treatment of hepatocellular carcinoma.

According to an aspect of the present invention, there is provided the use of a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs and optionally one or more pharmaceutically acceptable excipients in the manufacture of a formulation for treating hepatocellular carcinoma.

According to an aspect of the invention, there is provided a method of treating hepatocellular carcinoma wherein the method comprises administering a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs and optionally one or more pharmaceutically acceptable excipients.

BRIEF DESCRIPTION OF THE DRAWINGS:
A) Dissolution studies were carried out on Gelatin and HSA nanoparticles of Cisplatin.
Figure 1: Dissolution profiles of Gelatin and HSA nanoparticles of Cisplatin
Plain cisplatin exhibited quick dissolution in phosphate buffer saline (pH 7.4) within 4 hours due to its short half-life after intravenous administration.
In case of HSA nanoparticles, around 23% of Cisplatin was quickly released from the nanoparticles within 1 hour followed by a sustained release for a period of 96 hours. HSA-TG nanoparticle formulation exhibited a sustained release pattern for 72 hours.
In case of gelatin nanoparticles, sustained release profile for more than 72 hours was observed. Around 70% of the drug was released in 8 hours followed by a controlled release pattern beyond this point.
B) Sulforhodamine B assay
Figure 2: Sulforhodamine B assay for Gelatin nanoparticles
Plain cisplatin showed cytotoxicity on the liver cancer cell line causing disruption in the cancer cell structure and their shrinkage (GI50 = 3.6 µM).
Cisplatin gelatin nanoparticles exhibited cytotoxicity on the HepG2 cells, but the cytotoxicity was less as compared to pure drug (GI50 = 18.2 µM). This was attributed due to slow release of cisplatin from developed gelatin nanoparticles.
Blank nanoparticles did not exhibit any cytotoxicity (GI50 > 100 µM). C) Biodistribution studies on HSA and Gelatin nanoparticles of Cisplatin

Figure 3: Biodistribution studies of cisplatin gelatin nanoparticles in comparison to plain cisplatin
The concentration of cisplatin gelatin nanoparticles in the liver increased slowly till 48 hours; however, this concentration was found to reduce over 96 hours. Concurrently, plain cisplatin concentration was found to rise steadily in the lungs over a period of 96 hours. The initial high concentration of cisplatin gelatin nanoparticles in the liver was attributed to the enhanced permeability and retention (EPR) effect leading to rapid distribution in the liver followed by subsequent distribution in lungs. Some amount of cisplatin gelatin nanoparticles after i.v administration could reach the kidney and brain tissues over a period of 24 hours. However, over a time period of 96 hours, the concentration in the brain was found to reduce and increased in liver and lungs.

Figure 4 & Figure 5: Biodistribution studies on passively targeted and actively targeted cisplatin albumin nanoparticles
The concentration of cisplatin in case of non-targeted cisplatin albumin nanoparticles was maximum in case of liver and was seen to be 4 - 6 times higher as compared to other highly perfused organs. This phenomenon may be related to the preferential uptake of negatively charged albumin nanoparticles by the liver due to enhanced permeability and retention (EPR) effect. Also, the concentration of cisplatin was highest at 48 hours and was maintained throughout the time period of 96 hours in this study, due to slow diffusion of cisplatin from the albumin matrix. High liver concentration was achieved by the targeted galactosamine nanoparticles at all the time points and was significantly elevated as compared to all the other organs.

There was no significant difference between the cisplatin nanoparticle concentrations in the liver for non-targeted as well as the targeted formulations at 48 hours. However, this difference was significant for cisplatin concentration at 72 and 96 hours and it was seen that cisplatin was retained in the liver from galactosamine anchored nanoparticles to a greater extent as compared to the non-targeted albumin nanocarriers.

Figure 6: Biodistribution studies on TG-linked albumin nanocarriers
The highest concentration of cisplatin was attained in the liver with low concentrations in other organs. Concentration of cisplatin was maintained at high levels throughout the time period of 96 hours.

DETAILED DESCRIPTION OF THE INVENTION:
The main aim of anticancer therapy is to ensure specific drug action at the target site/tissue/organ along with reduced distribution to other non-specific organs.

Target oriented drug delivery systems can provide maximum therapeutic benefit through controlled and predetermined release rate kinetics, prevent drug degradation or inactivation during transit to target sites and prevent the body from adverse reactions. For drugs that possess low therapeutic index, these systems can provide effective treatment at low concentrations.

Nanoparticles due to their colloidal nature, offer numerous advantages over conventional treatment strategies. Small sized particles possess large surface area and hence increase the dissolution properties of antineoplastic agents with poor solubility. Nanoparticles can be tailored to reach specific tumor sites by modulation of their size, surface characteristics, and particle charge.

Delivery of anticancer agents in the form of nanoparticles also ensures reduction in their therapeutic dosage and frequency of administration due to the controlled release properties of the nanosystems.

Additionally, nanoparticulate drug delivery systems reduce the untoward toxicities associated with antitumor agents and prevent their degradation. Site specific targeting is possible with nanoparticles after attachment of certain ligands, antibodies or carriers on the surface by physical adsorption or covalent attachment.

Higher drug loading can be achieved in case of nanocarriers, which is a prime factor for any particulate drug delivery system. Nanoparticles possess the ability to increase the absorption of anticancer agents from the body upon administration, thus causing a concomitant rise in their bioavailability.

These particles can act at cellular levels and can be endocytosed/phagocytosed by cells, with resulting cell internalization of the encapsulated drug. Also, the small size of colloidal carriers makes them suitable for parenteral delivery of drugs. Irritation potential at the site of injection can be minimized with nanocarriers as compared to conventional therapy. Nanoparticles are biocompatible and non-immunogenic and can entrap hydrophilic and hydrophobic moieties, proteins, vaccines and biological macromolecules.

Nanoparticles can be prepared using both non-biodegradable and biodegradable polymers. However, the later are preferred due to their biocompatibility, biodegradability, low cost and safety. These particles can be delivered by various routes of administration like nasal, oral, dermal and ocular, apart from parenteral route.

Biodegradable polymeric nanocarriers are favorable due to their excellent biocompatibility, biodegradability and low toxic potential. These biodegradable polymers include, but are not limited to, natural polymers like gelatin, human serum albumin, sodium alginate, agarose, casein, zein, chitosan, glycol chitosan, N, N trimethyl chitosan, starch, cellulose and wheat gluten.

Human Serum Albumin (HSA) is the most important soluble protein in human body, which is synthesized in the liver. It is the primary protein in human body controlling the osmotic pressure of serum. Other functions imparted by albumin include binding of toxic products and their transport to the liver, distribution and transport of metal ions and other exogenous components. Albumin is an acidic protein with an isoelectric point of 4.7. This protein is stable in the pH range of 4-9 and exhibits solubility in water and other aqueous solutions. Albumin can be heated at a temperature of around 60°C, without any change in the protein structure. All these properties along with the preferential tumor uptake, biodegradability, non-toxicity and property to accumulate at inflamed tissues, makes it an ideal polymer for cancer drug delivery.

Gelatin, a natural macromolecule is commonly employed as a pharmaceutical adjuvant and an encapsulating drug material. Advantages of gelatin for drug delivery include its inexpensiveness, ability to be sterilized, free from contamination with pyrogens and low antigenicity.

Preferably, the polymers may be present in an amount ranging from about 0.1% to about 35% by weight of the composition.

According to the present invention, human serum albumin and gelatin are the preferable polymers that have been employed in the pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs.

Preferably, the pharmaceutical composition, according to the present invention, comprises polymeric nanoparticles of anticancer drugs.

According to one aspect of the present invention, the pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs have been engineered to have an affinity for target tissues through passive or active targeting mechanisms.
According to another aspect of the present invention, the pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs deliver the said anticancer drugs to the target site of action as well as attempt to decrease or avoid the side effects.

According to another aspect of the present invention, the pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs exhibits prolonged release.

According to another aspect of the present invention, the pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs which have a reduced dose.

Anticancer drugs according to the present invention include, but are not limited to alkylating agents, antitumor antibiotics, anti-microtubule agents, DNA linking agents, bisphosphonates, topoisomerase inhibitors, antimetabolites, biological agents and nucleoside analogues.

Suitable alkylating agents according to the present invention include one or more but are not limited to mechlorethamine, mustine, cyclophosphamide, nitrosoureas, tetrazine, azidine, melphalan, chlorambucil, busulfan, uramustine, bendamustine, ifosfamide, lomustine, semustine, carmustine, fotemustine, streptozotocin, thiotepa, dacarbazine, procarbazine, mitomycin, mitozolomide, temozolomide and the like or combinations thereof.

DNA linking agents include one or more but are not limited to cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin and the like or combinations thereof.

Suitable antitumor antibiotics and topoisomerase inhibitors according to the present invention include one or more but are not limited to bleomycin, daunorubicin, epirubicin, doxorubicin, idarubicin, mitoxantrone, mitomycin, actinomycin, pirarubicin and the like or combinations thereof.

Suitable antimetabolites according to the present invention include one or more but are not limited to fluorouracil, capecitabine, asparaginase, cladribine, nelarabine, decitabine, clofarabine, pentostatin, methotrexate, pemetrexed, raltitrexed, asparaginase, thioguanine, mercaptopurine and the like or combinations thereof.

Suitable biological agents according to the present invention include one or more but are not limited to bevacizumab, cetuximab, regorafenib, ibrutinib, axitinib, crizotinib, dabrafenib, ponatinib, afatinib, dasatinib, nilotinib ruxolitinib, trametinib, erlotinib, sorafenib, gefitinib, imatinib, interferon, lapatinib, ipilimumab, denosumab, panitumumab, rituximab, sunitinib, emsirolimus, trastuzumab and the like or combinations thereof.

Suitable nucleoside analogues according to the present invention include one or more but are not limited to didanosine, vidarabine, abacavir, acyclovir, entecavir, cytarabine, lamivudine, emtricitabine, stavudine, telbivudine, idoxuridine, trifluridine, zidovudine and the like or combinations thereof.

Other anticancer agents according to the present invention include one or more but are not limited to mesna, octreotide, tamoxifen, anastrozole, amifostine, bexarotene, bicalutamide, buserelin, exemestane, flutamide, folinic acid, cyproterone, degarelix, folinic acid, fulvestrant, goserelin, lanreotide, letrozole, medroxyprogesterone, megestrol, stilbestrol, thalidomide and the like or combinations thereof.

In one embodiment, the pharmaceutical composition according to the present invention comprises human serum albumin nanoparticles as well as gelatin nanoparticles of cisplatin. The term “cisplatin” is used in broad sense to include not only “cisplatin” per se but also its pharmaceutically acceptable derivatives thereof. Suitable derivatives include pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, pharmaceutically acceptable isomers, pharmaceutically acceptable esters, pharmaceutically acceptable anhydrates, pharmaceutically acceptable enantiomers, pharmaceutically acceptable polymorphs, pharmaceutically acceptable prodrugs, pharmaceutically acceptable tautomers and/or pharmaceutically acceptable complexes thereof.
The pharmaceutical composition, according to the present invention, comprises polymeric nanoparticles of anticancer drugs which can be administered by varying routes of administration such as but not limited to oral, parenteral (subcutaneous, intravenous, intramuscular, intradermal, intraperitoneal) or topical or localized (directly at the site)and the like.

The pharmaceutical composition, according to the present invention, comprises albumin as well as gelatin nanoparticles of anticancer drugs, which can be provided as but not limited to oral, parenteral or topical dosage forms, localized and the like.

The term "pharmaceutical composition" includes oral dosage forms, such as but not limited to, tablets, soft gelatin capsule, capsules (filled with powders, powders for reconstitution, pellets, beads, mini-tablets, pills, micro-pellets, small tablet units, MUPS, disintegrating tablets, dispersible tablets, granules, sprinkles, microspheres and multiparticulates), sachets (filled with powders, pellets, beads, mini-tablets, pills, micro-pellets, small tablet units, MUPS, disintegrating tablets, dispersible tablets, granules, sprinkles microspheres and multiparticulates) and sprinkles, however, other dosage forms such as liquid dosage forms (liquids, liquid dispersions, suspensions, solutions, emulsions, sprays, spot-on), micro-formulations, nanoformulations may also be envisaged under the ambit of the invention.

The term "pharmaceutical composition" includes parenteral dosage forms such as liquid dosage forms (liquids, liquid dispersions, suspensions, solutions, emulsions, powders for reconstitution), gels, bolus, depots, implants (rods, capsules, rings) biodegradable or non-biodegradable microparticles/microspheres etc. may also be envisaged under the ambit of the invention.

The term "pharmaceutical composition" includes topical dosage forms, such as but not limited to, sprays, solutions, suspensions, ointments, drops, in-situ gel, aerosols, ointments, microspheres, creams, gels, patches, films and the like.

In one embodiment, the pharmaceutical composition of the present invention comprises polymeric nanoparticles of anticancer drugs, in the form of controlled release formulations, lyophilized formulations, delayed release formulations, timed release formulations, extended release formulations, pulsatile release formulations, and mixed immediate release and controlled release formulations.

Preferably, the pharmaceutical compositions of the present invention comprising polymeric nanoparticles of anticancer drugs are provided in parenteral dosage forms.

Accordingly, the pharmaceutical composition of the present invention comprising polymeric nanoparticles of anticancer drugs is provided in a lyophilized parenteral dosage form, these dosage forms are to be dissolved or reconstituted in a suitable solvent prior to administration.

Suitable solvents for reconstitution include but are not limited to purified water for injection, sterile saline solution, dextrose solution, Ringer’s solution and the like.

The present invention thus provides a pharmaceutical composition comprising polymeric nanoparticles of the anticancer drugs wherein such nanoparticles have an average particle size of less than or equal to about 2000 nm, preferably less than or equal to about 1000 nm.

The term average particle size as used herein refers to the average diameter of the particles.

The term “particles” as used herein refers to an individual particle of the polymeric nanoparticle comprising anticancer drug(s), or the polymeric nanoparticles comprising anticancer drug(s).

The particles of the present invention can be obtained by any of the process such as but not limited to milling, precipitation, homogenization, high pressure homogenization, spray-freeze drying, supercritical fluid technology, double emulsion/solvent evaporation, desolvation, PRINT (Particle replication in non-wetting templates), thermal condensation, ultrasonication, interfacial polymerisation, spray drying and combinations thereof. Preferably, the particles of the present invention are prepared by the process of desolvation.

Accordingly, the process of desolvation can comprise dissolving the anticancer drug in aqueous solution of human serum albumin or gelatin and adding a desolvating agent for formation of nanoprecipitates, which are further cross linked and then neutralized. Further, the purified nanoparticles are optionally freeze dried.

Suitable desolvating agents according to the present invention include one or more but are not limited to acetone, ethanol or isopropanol and the like or combinations thereof.

Suitable cross linkers for stabilization of human serum albumin or gelatin nanoparticles according to the present invention include one or more but are not limited to bifunctional aldehydes like glutaraldehyde and formaldehyde, carbodiimides (EDC), enzymes such as transglutaminase or thermal processes and the like or combinations thereof.

Preferably, one or more cross linkers may be present in an amount ranging from about 0.1% to about 20% by weight of the total composition.

Suitable neutralizers according to the present invention include one or more but are not limited to glycine, lysine, sodium metabisulphite, sodium sulphate and the like or combinations thereof.

Preferably, one or more neutralizers may be present in an amount ranging from about 0.1% to about 20% by weight of the total composition.

Suitable cryoprotectants according to the present invention include one or more but are not limited to sucrose, trehalose, dextrose, mannose, glycine, glucose, galactose, lactose and the like or combinations thereof.
Preferably, one or more cryoprotectants may be present in an amount ranging from about 0.1% to about 20% by weight of the total composition.

Suitable pH adjusting agents or buffering agents that may be used in the pharmaceutical composition include but are not limited to hydrochloric acid, acetic acid, citric acid, tartaric acid, propionic acid, sodium hydroxide, sodium phosphate, ammonia solution, triethanolamine, sodium borate, sodium carbonate, sodium acetate, potassium hydroxide and the like or combinations thereof.

Preferably, one or more buffering agent may be present in an amount ranging from about 0.1% to about 10% by weight of the total composition.

Suitable solvents/co-solvents, solubilizer or vehicles, that may be employed, in the pharmaceutical composition include, but are not limited to, dichloromethane, acetonitrile, ethyl acetate, acetone, propylene carbonate, water, glycerine, coconut fatty acid diethanolamide, medium and/or long chain fatty acids or glycerides, monoglycerides, diglycerides, triglycerides, structured triglycerides, soyabean oil, peanut oil, corn oil, corn oil mono glycerides, corn oil di glycerides, corn oil triglycerides, polyethylene glycol, caprylocaproylmacroglycerides, caproyl 90, propylene glycol, polyoxyethylenesorbitan fatty acid esters, polyoxyethylene castor oil derivatives, castor oil, cottonseed oil, olive oil, safflower oil, peppermint oil, coconut oil, palm seed oil, beeswax, oleic acid, methanol, ethanol, isopropyl alcohol, butanol, acetone, methyl isobutyl ketone, methyl ethyl ketone or combinations thereof.

The present invention also provides a method of treating hepatocellular carcinoma by administering a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs.

The present invention also provides the use in the treatment of hepatocellular carcinoma by administering pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs.
It may be well acknowledged to a person skilled in the art that the said pharmaceutical composition, according to the present invention, may further comprise one or more active in particular other anticancer drugs, such as, but are not limited to, alkylating agents, anti-metabolites, anti-microtubule agents, topoisomerase inhibitors or antitumor antibiotics, DNA linking agents, biological agents and bisphosphonates.

The following examples are for the purpose of illustration of invention only and are not intended in any way to limit the scope of the present invention.

Example 1:
a) Bolus injection of HSA nanoparticles of Cisplatin
Sr. No Ingredients Quantity
1 Human serum albumin 100 - 300 mg (10 ml )
2 Cisplatin 5 - 12 mg
3 Absolute ethanol 10 - 20 ml
4 Glutaraldehyde 50 - 200 µl (8 %w/v)
5 Glycine 40 - 100 mg
6 Vehicle (10 mM NaCl solution) 10 ml

Process:
1. Cisplatin was added to the aqueous human serum albumin solution while stirring after pH adjustment. .
2. Ethanol was added to the solution obtained in step (1) to cause precipitation of the protein molecules leading to the formation of nanoparticles.
3. The nanoparticles obtained in step (2) were cross linked with glutaraldehyde under stirring.
4. Glycine was added under stirring to the nanoparticles obtained in step (3) to remove excess glutaraldehyde.
5. The nanoparticle solution obtained in step (4) was ultracentrifuged, redispersed in phosphate buffered saline (PBS), lyophilized and provided with a sterile solution for reconstitution.
b) Bolus injection of albumin nanoparticles of Cisplatin anchored with galactosamine for liver specific targeting
Sr. No Ingredients Quantity
1 Human serum albumin 100 - 300 mg (10 ml )
2 Cisplatin 5 - 12 mg
3 Absolute ethanol 10 - 20 ml
4 Glutaraldehyde 50 - 200 µl (8 %w/v)
5 Glycine 40 - 100 mg
6 Galactosamine 10 - 20 mg
7 EDC (carbodiimide) 10 - 20 mg
8 Vehicle (10 mM NaCl solution) 10 ml

Process:
1. Cisplatin was added to the aqueous human serum albumin solution while stirring after pH adjustment.
2. Ethanol was added to the solution obtained in step (1) to cause precipitation of the protein molecules leading to the formation of nanoparticles.
3. The nanoparticles obtained in step (2) were cross linked with glutaraldehyde under stirring.
4. Glycine was added under stirring to the nanoparticles obtained in step (3) for removal of excess glutaraldehyde.
5. The nanoparticle solution obtained in step (4) was ultracentrifuged, redispersed in chilled phosphate buffered saline (PBS) containing galactosamine and EDC, lyophilized and provided with a sterile solution for reconstitution.

c) Bolus injection (IV) of albumin nanoparticles of Cisplatin anchored with Transglutaminase
Sr. No Ingredients Quantity
1 Human serum albumin 100 - 300 mg (10 ml )
2 Cisplatin 5 - 12 mg
3 Absolute ethanol 10 - 20 ml
4 Transglutaminase enzyme 50 - 100 mg
6 Vehicle (10 mM NaCl solution) 10 ml

Process:
1. Cisplatin was added to aqueous human serum albumin solution under stirring after pH adjustment.
2. Ethanol was added to the solution obtained in step (1) to cause precipitation of the protein molecules leading to the formation of nanoparticles.
3. The nanoparticles obtained in step (2)were cross linked with transglutaminase enzyme under stirring
4. The nanoparticle solution obtained in step (4) was ultracentrifuged, redispersed in phosphate buffered saline (PBS), lyophilized and provided with a sterile solution for reconstitution.

d) Bolus injection of gelatin nanoparticles of Cisplatin
Sr. No Ingredients Quantity
1 Injectable grade Gelatin 100 - 300 mg (10 ml )
2 Cisplatin 5 - 12 mg
3 Acetone 10 - 30 ml
4 Glutaraldehyde 50 - 200 µl (8 %w/v)
Glycine 40 - 100 mg
6 Vehicle (10 mM NaCl solution) 10 ml

Process:
1. Gelatin was dissolved in water and acetone was added.
2. The supernatant containing low molecular weight (LMW) fraction of gelatin discarded by decantation. The remaining high molecular weight (HMW) sediment was re-dissolved in equal volume of distilled water.
3. The pH of gelatin solution obtained in step (2) was adjusted and cisplatin was dissolved in it and gelatin was desolvated again by addition of acetone leading to the formation of nanoparticles.
4. Glutaraldehyde was added to the nanoparticles obtained in step (3) under stirring.
5. Glycine was added under stirring to the nanoparticles obtained in step (4) for removing excess glutaraldehyde.
6. The nanoparticle solution obtained in step (5) was ultracentrifuged, redispersed in phosphate buffered saline (PBS), lyophilized and provided with a sterile solution for reconstitution.

It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the spirit of the invention. Thus, it should be understood that although the present invention has been specifically disclosed by the preferred embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and such modifications and variations are considered to be falling within the scope of the invention.

It is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of “including,” “comprising,” or “having” and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items.

It must be noted that, as used in this specification and the appended claims, the singular forms "a," "an" and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to "an excipient" includes a single excipient as well as two or more different excipients, and the like. ,CLAIMS:WE CLAIM:

1. A pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs or any of their pharmaceutically acceptable salts, solvates, complexes, hydrates, isomers, esters, tautomers, anhydrates, enantiomers, polymorphs, prodrugs or derivatives thereof and one or more pharmaceutically acceptable excipients.
2. A pharmaceutical composition according to claim 1, wherein the polymer is biodegradable, non-biodegradable or combination thereof.
3. A pharmaceutical composition according to claim 1, wherein the anticancer drug is DNA linking agent.
4. A pharmaceutical composition according to claim 1, wherein the DNA linking agent is cisplatin.
5. A pharmaceutical composition according to preceding claims wherein polymer is a biodegradable polymer selected from Human Serum Albumin (HSA) or Gelatin or a combination thereof.
6. A pharmaceutical composition according to any preceding claim, wherein the nanoparticles have an average particle size of less than or equal to about 2000 nm.
7. A pharmaceutical composition according to any preceding claims for parenteral administration.
8. A pharmaceutical composition according to claim 7, wherein the dosage form for parenteral administration is in the form of a liquid, gel, bolus, depots, implant, biodegradable or non-biodegradable microparticles, microspheres.
9. A pharmaceutical composition according to any preceding claims wherein the pharmaceutically acceptable excipients are selected from desolvating agents, cross linkers, neutralisers, cryoprotectants, buffering agents, ligands and solvents.
10. A pharmaceutical composition according to any preceding claim, further comprising at least one additional active ingredient selected from alkylating agents, anti-metabolites, anti-microtubule agents, topoisomerase inhibitors or antitumor antibiotics, biological agents or bisphosphonates.
11. A process for preparing a pharmaceutical composition comprising polymeric nanoparticles of anticancer drugs, said process comprising dissolving the anticancer drug in aqueous solution of human serum albumin or gelatin and adding a desolvating agent for formation of nanoprecipitates, cross linking and neutralization of the formed nanoparticles with other pharmaceutically acceptable excipients, optionally freeze drying the purified nanoparticles and processing into the desired dosage form.
12. A pharmaceutical composition according to any one of claims 1 to 11 for use in the treatment of hepatocellular carcinoma.
13. A pharmaceutical composition as described herein with reference to the examples.