WO2016167730A1

WO2016167730A1 - Nanomicelles for the treatment of cancer

Info

Publication number: WO2016167730A1
Application number: PCT/TR2015/000320
Authority: WO
Inventors: Oğuz Aslan ÖZEN; Mustafa ÖZGÜL; Muhammed AYDIN
Original assignee: Özen Oğuz Aslan; Özgül Mustafa; Aydin Muhammed
Priority date: 2015-04-13
Filing date: 2015-09-21
Publication date: 2016-10-20

Abstract

The invention is about a cancer obliterator that contains anticancer drug enriched by H5WYG peptides containing several histidine with a property escaping from endosomes; and solves the problem of the vasoactive intestinal peptides and drug-loaded nanoparticles taken into the cell (which play a role in endocytose process and specifically bind to the receptors on cancer cell membranes) not being released into the cytosol by being held by endosomes.

Description

Instruction Book

NANOMICELLES FOR THE TREATMENT OF CANCER

Technical Area

The invention is about a cancer obliterator that contains anticancer drug enriched by H5WYG peptides containing several histidine amino acids with a property escaping from endosomes; and solves the problem of the vasoactive intestinal peptides and drug-loaded nano-particles taken into the cell (which play a role in endocytose process and specifically bind to the receptors on cancer cell membranes) not being released into the cytosol by being held by endosomes.

Current state of the technique

The new developed strategies provide that anticancer drugs systematically applied go to the target region selectively, and target the drug accumulation in the sick region and minimize the cytotoxic effect of anticancer drug on the healthy cells. Polymeric particles, liposomes, micelles, nano-emulsions and similar methods are utilized in the pharmaceutically transport systems of anticancer drugs to the target region. Nowadays, there are drug transport approaches to the various tumors with the methods of passive targeting and active targeting.

The one of the most used and known method in the current technique is passive targeting. There are two characteristics of solid tumors in the studies about this method;

• Highly irregular tumor blood vessels (Peer et al., 2007)

· Dysfunctional lymphatic drainage (Matsumura and Maeda, 1986)

The junctions between the endothelial cell lines of tumor blood vessels disappear, in conclusion, large molecules and particles (in size of 10— 500 nm) can pass into the tumoral area because of the high the permeability of tumor vessels (Serpe, 2006; Torchilin, 2010) (Figure 1).

There are tight junctions between endothelial cells of the normal capillaries as shown in Figure 1. it shows tumor vascular structure with high capacity of permeability by the decrease of the tight junctions between the endothelial cells constituting the structure of capillary feeding the tumor cells. The drug carrier nanoparticles (NP) in the circulatory system reach to the tumor cells by leaking through the decreased tight junctions of the endothelial cells thanks to the 'EPR effect (Enhanced Permeability and Retention effect)'.

In passive drug carrying method, the EPR effect provides the accumulation of the drug in the interstitial region of the tumor by leaking through the tumor vessels with nanocarriers. In addition to this, the degenerate lymphatic clearance in tumors (clearance) provides the drug loaded nanoparticles to accumulate in tumor interstitial region. By this reason, release of encapsulated drug into the interstitial area or internalization of encapsulated drug by cancer cells can occur via endocytosis/phagocytosis. In conclusion, drug accumulation increases in the targeted tumor region and anticancer efficiency of chemotherapeutic agents can be increased successfully. However, most of the tumors stay out of mononuclear phagocytosis system. Therefore, by this type of targeting, the circulation half-life of drug loaded nano particles is limited with the elimination by the macrophages (Koo et al, 2005).

Consequently, the sterically stabilized carriers coated with hydrophilic polymer (Polyethylene glycol) (PEG), poloxamin, poloxamer and polysaccharide) with long circulation half-life have been developed. Phagocytosing developed nanocarriers by immune cells digesting and destroying foreigner organism and molecules in the body (phagocytes) through decreasing the binding of plasma membrane proteins. For example, it has been shown that Doxil®-PEG-coated liposomes approved by FDA (Food and Drug administration) has -275 times longer half-life and accumulates more in the tumoral region when compared with free doxorubicin (Gabizon, 2001). The other technique in current methods is the active targeting. Passive targeting methods are used widely in the clinical treatments. However, active targeting is developed for providing more selectivity and more drug intake to the tumoral regions, and also preventing the elimination of the drugs from the cells in case of multi-drug resistance. These developed strategies include the drug carrier structures which are coated with ligands binding to the receptors over-expressing on tumor cell surface (Allen, 2002). Monoclonal antibodies and their fragments regulate the cancer cell proliferations by binding to the receptors. For example, trastuzumab is utilized in breast cancer treatment due to binding to the human epidermal growth factor receptor 2 (EGFR-2) (Danhier et al., 2010).

Radioimmunotherapeutic agents; therapeutic molecules and antibody are localized on the tumor surface by being combined. An important feature of the targeted nanocarriers is carrying the cytotoxic drugs by keeping a targeting ligand on the surface of nano-sized drug carrying system (Koo et al. , 2005; Lammers et al., 2008; Peer et al., 2007). Antibodies, peptides, aptamers (short single-strand DNA or Ribonucleic acid oligonucleotides), vitamins and carbohydrates are being investigated as the targeting parts for the nano-carriers (Serpe, 2006; Lammers et al., 2008). Although current clinical evidences are insufficient for the active targeting strategy, pre-clinical data obtained from animal studies for the last ten years are promising. Studies have targeted the nano-carriers' receptors having only internalized characteristic and anti-cancer efficiency was shown to be increased significantly (Marcucci et al., 2004; Lammers et al.,2008; Kirpotin et al., 2006). The drug activity was provided to be increased by receptor-mediated endocytosis of nanocarriers and larger drug accumulation is prevented in tumor interstitium. These findings have provided that internalized receptors on cell surface (eg. folate receptor and vasoactive intestinal peptide (VIP) receptors) are selected as target regions in the drug transport systems (Lee and Low., 1994; Leamon et al., 2003; Park et al., 2002; Krishnadas, 2004; Koo et al., 2011 ; Onyuksel et al., 2009). In addition, targeting strategies have been investigated through protein transduction domain mediation (eg. TAT peptides) and cell penetrating peptides. TAT peptides can be benefited for internalizing to the cytosol without interacting with surface receptors (Peer et al., 2007; Marcucci and Lefoulon., 2004; Torchilin et al., 2001). However, these peptides are not cell specific and by this reason their usage is limited.

Another method in the current technique is target-specific sterically stabilized phospholipid micelles. In this method, drug loaded nanoparticles may constitute of one or more materials. These are natural or synthetic polymers and lipids. Numerous nanocarriers have been developed over the years. These developed nanocarriers are polymeric nanospheres, nanoemulsions, dendrimers, nanocapsules, liposomes and micelles. Nanovectors must be smaller than 200 nm, must have a high drug loading capacity, must be biodegradable and have biocompatibility, and they also must have a long circulation period in the body and target the sick areas and show their effects in the unhealthy regions to be successfully used in anticancer therapy in clinic (Torchilin, 2010).

Sterically stabilized phospholipid micelles (SSM) consist of PEGylated phospholipid (DSPE-PEG 2000). SSMs have the necessary characteristics to be used for clinical anticancer treatment. Phospholipid is an amphiphilic molecule which has nonpolar hydrocarbon domain and polar phosphate head domain. When DSPE-PEG 2000 is added to the aqueous medium, lipid molecules come together (self-aggregate) as a result of the specific characteristic of molecule and by this way the undesired hydrophobic interactions are minimized. T/TR2015/000320

DSPE-PEG 2000 molecule takes the shape of a cone because of Gaussian random coil of PEGs connected to the polar head domain of phospholipid. It should be over certain lipid concentrations so the formation of appropriate micelle in aqueous mediums can occur. This definite lipid concentration is known as critical micelle concentration. When SSMs ranging between 0.5 and 1.0 μΜ are applied into the body in relative low critical micelle concentrations, high thermodynamic stability and negligible disintegration can occur in micelles (Ashok et al., 2004).

Studies have shown that anticancer drugs such as paclitaxel, camptothecin that include hydrophobic molecules, dissolve in SSM (Lim et al., 2011 ; Sezgin et al., 2006; Koo et al., 2005; Onyuksel et al., 1999). Hydrophobic molecules move towards the micelle nucleus in water and the hydrophilic molecules hold on to micelle surface (Torchilin, 2010). Phospholipid micelles can be targeted to the cancerous area via passive or active targeting mechanism. SSMs in approximately 15 nm size can be targeted to the cancer tissue passively. In addition to this, SSMs can be also be targeted to the tumor region by adding specific ligands such as peptide to the distal end of PEG chain (Onyuksel et al., 2009).

Another method of the current technique is H5WYG peptide method. Success in nanomedicine and gene therapy application is highly related to the development of vectors for applying gene or treating agents to the target cells with minimum toxicity in a selective and efficient way (Nishikawa et al., 2001 ; Tokatlian and Segura 2010). The internalization of nanoparticles into the cell by endocytosis and the endosomal escape causing destabilization in endosomal membrane via endosomolitic peptides are shown in Figure 2. Early endosomes consisting of vesicles containing therapeutics are formed by cell surfaces. Late endosomes are the last cluster of a series of stages prior to interactions with lysosome and it consists of early endosome materials. Lysosomes are the last part of the endocytic pathway and there are hydrolytic enzymes breaking up the content of the late endosomes within. For this reason, the escape of the therapeutics from the endosome is necessary, before the lysosome-mediated disintegration of the therapeutics (Carlee et al., 2011). Various methods have been tested for early secretion of therapeutic cargos into the cytosol in endosomal pathway. These methods are based on the description of endosomal escape mechanisms such as pore formation, pH buffering effect and structural changes increasing endosomal escape. Viral proteins, bacterial proteins and especially synthetic biomimetics are used as endosomal releasing agents in the nucleic acid and protein delivery system (Amir et al., 201 1). Endosomal pathway is one of the cellular uptake mechanisms. Endosomal pathway includes vesicles named as endosome at about 5 pH and occurs from early to late endosomes as one-way path before the merging with intracellular organelles known as lysosomes containing digestive enzymes (Gruenberg and Goot, 2006). Particles are taken into the cells via endosomes through endosomal pathway and in the last step, it reaches to the lysosome and active enzymatic digestion process occurs. This situation causes limitation in the delivery of therapeutic agents into the cell. Thus, although there are many promising and potential compounds in in vitro application, it cannot be applied in vivo because of bioavailability problems. Until now, many ways have been tried for direct transport into the cell by escaping various macromolecules from the endosomal pathways and protecting them from the cellular degradation (Amir et al., 201 1 ).

Endosomal escape is provided by creating destabilization on the endosomal membrane via endosomolitic peptides (Marsh and Helenius, 1989; Horth et al., 1991). Although studies have shown that H5WYG (GLFHAIAHFIHGGWHGUHGWYGGGC) peptides permeabilize the cell membrane in mildly acidic pH, not in neutral pH, however the peptide was shown to make serious structural changes in pH between 7.0 and 6.0 through absorbance, fluorescence and circular dichroism spectrums (Murata et al., 1992; Midoux et al., 1998).

Midoux et al. (1998) have shown that H5WYG peptide permeabilizes the cell membrane in a great efficiency in pH 6.4 in living cells in their cell permeabilization study. H5WYG peptide is not active in neutral peptide however, H5WYG peptide was shown to permeabilize 50% of cells in pH 6.8 at 20 °C in 10 minutes, through the flow cytometry. Nano-sized particles modified with histidine-rich H5WYG peptide was shown to prevent the degradation in the endolysosome and disperse in the cytosol in 4 hours by providing endosomal escape in in vitro studies (Carlee et al., 2011 ). Besides, osmotic swelling and membrane destabilization occur by the 'proton sponge' mechanism through protonated imidazole parts of endosomolitic peptide (Behr et al., 1997).

Another known method among current methods is anticancer drugs placed in nanomiceiie structure. Nigella sativa is a spice in the east which is known as black cumin. The essential oils of Nigella sativa consist of thymoquinone, thymohydroquinone, dithymoquinone, and thymol by the High Performance Liquid Chromatography (HPLC) analysis (Gosheh et al., 1998). Thymoquinone is the most bioactive compound in Nigella sativa (Motaghed et al., 2013). Pharmacological and therapeutic effects of thymoquinone have been shown in studies with disease models in vivo and in vitro (Butt and Sultan, 2010; llaiyaraja and Khanum, 2010; Chehl et al., 2009; Motaghed et al., 2013). Thymoquinone shows anti-cancer characteristic regulating p53, p73, STAT3, NF-KB, PPAR-γ and reactive oxygen products (Gali-Muhtasib et al., 2004; Gali-Muhtasib et al., 2008; Alhosin et al., 2010; Woo et al., 2011 ; Li et al., 2010; El-Najjar et al., 2010). Thymoquinone has anti-oxidant, anti-inflammatory and anti-cancer properties (Gali- Muhtasib vd., 2004; Badary vd., 1999; Badary vd., 2001). It was determined to suppress proliferation of various tumor cells such as pancreatic carcinoma, breast adenocarcinoma, colorectal carcinoma, osteosarcoma (Gali-Muhtasib et al., 2004; Gali-Muhtasib et al., 2008; Roepke et al., 2007; Shoieb et al., 20031 ; El-Mahdy et al., 2005; Tan et al., 2006).

Moreover, thymoquinone was shown not to have a toxic effect on the healthy cells in in vivo and ex vivo experimental models (Worthen et al., 1998; Attoub et al., 2012). Thymoquinone triggers p53-dependent and independent apoptotic pathways and it inhibits the MCF-7/DOX doxorubicin resistant breast cancer cells in the stage of G2/M and increases the p53 and p21 proteins (Arafa vd., 201 1). Thymoquinone suppresses DNA synthesis and cell cycle, and as a result inhibits the tumour growth (Worthen et al., 1998; Shoieb et al., 2003). Woo et al., (2013) found that thymoquinone inhibits XIAP (X-linked inhibitor of apoptosis protein), survivin, Bcl-xL and Bcl-2 anti-apoptotic gene products and it suppresses the tumor growth in breast cancer cells and breast cancer xenograft mice model. Arafa et al., (201 1) showed that thymoquinone induces the apoptosis of breast carcinoma cells by providing the up regulation of Bax protein and down regulation of Bcl-2 protein.

IC_5C value (50% inhibitor concentration) of thymoquinone was found to be approximately 25 μΜ for MCF-7 breast cancer cell line (Motaghed et al., 2013). Besides, thymoquinone was shown to inhibit the proliferation of breast cancer cell line at the end of the 72 hours when minimal dose was 25 μΜ and was shown to arrest in S phase in cell cycle analysis. When applying 100 μΜ thymoquinone to the MCF-7 cell line, decreasing cell viability by 49% was shown by flow cytometry at the end of 24 hours (Motaghed et al., 2013). Consequently, results of the studies show that thymoquinone has important anti-cancer activity mechanisms for breast cancer. However, developed drug distribution strategies have to be improved to use this promising natural substance in the clinics in the future.

Another method known in this technique is using thymoquinone loaded nanoparticle. In Shah's et al. (2010) study, PHA-mPEG copolymeric nanoparticles loaded with thymoquinone at the size of averagely 1 12-162 nm have been generated and shown to be biocompatible in in vitro cytotoxicity experiments. In another study, thymoquinone-loaded micelle-structured poly(D, L tactide- co-glycolide) (PLGA) nanoparticle in the size of 200 nm was shown to inhibit the growth of MDA-MB-231 cell line (Ganea et al., 2010).

Thymoquinone loaded nanoparticle in size of 150-200 nm has been shown to inhibit the activation of NF-kB on MCF-7 human breast cancer cell line, PC-3 human prostate cancer cell line, U266 multiple myeloma cell line, and HCT166 human colon cancer cell line, and suppress the expression of cyclin D1 , matrix metalloproteinase (MMP)-9, vascular endothelial growth factor (VEGF) (Ravindran et al 2010). Liposome loaded with thymoquinone in approximately 100 nm diameter suppresses the proliferation in MCF-7 and T47D breast cancer cell line and it has very little toxic effect on healthy periodontal ligament fibroblast (Odeh et al., 2012). When the products obtained from anticancer drug studies are examined, although there are products having passive targeting systems of nanoparticle structures loaded with anti-cancer drug, the problem by which nanoparticles taken by endosomes thus anticancer drug not to release into the cytosol is the biggest disadvantage of the current anticancer drugs. Moreover, the most of anticancer drugs have lipophilic characteristic, they can cause damage to almost all healthy cells. For this reason, there are serious side effects of anticancer drugs. The development of new drug delivery strategies featured passive targeting, active targeting and endosomal escape are required.

Although, there are studies demonstrating passive targeting systems with non-toxic effects of the nanoparticles structure loaded with thymoquinone; developed active targeting featured drug delivery strategies needs to be developed.

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Brief explanation of the invention

The current invention is about a cancer obliterator that meets the above mentioned requirements, removes all the disadvantages and additionally brings some advantages.

The primary objective of the invention is, a cancer obliterator that contains anticancer drug enriched by H5WYG peptides containing several histidine amino acids with a property escaping from endosomes; and solves the problem of the vasoactive intestinal peptides and drug-loaded nanoparticles taken into the cell (which play a role in endocytose process and specifically bind to the receptors on cancer cell membranes) not being released into the cytosol by being held by endosomes. It is a product that will be produced for the first time in sterically stabilized micelles and one of its most important advantages is that is does not cause the side effects that occur with the other anti-cancer drugs.

Another objective of the invention is that the newly developed anticancer drug-loaded SSMs are in the appropriate nano-size and biodegradable to reach the cancer region by passive targeting. It also contains the receptor-specific peptide thus it will fill an important gap in this area by its active targeting feature and has the capability of being a leading product to create a strong anticancer effect.

The figures assisting in understanding of the invention

The subject of our application; the developed structure of "cancer obliterator" and other matters in current technique are shown in the attached figures. These are; Figure 1 shows the tumor vascular structure with the feature of high permeability by the reduction of tight junctions between endothelial cells forming the capillary structure that is feeding the tumor cells.

Figure 2 shows endosomal escape by destabilizing in endosomal membrane via endosomolitic peptides and the internalization of nanoparticles into the cell by endocytosis.

Figure 3 Schematic illustration of Anticancer drug-loaded SSM (Sterically stabilized micelle). Figure 4 Schematic illustration of Anticancer drug-loaded SSMM (Sterically stabilized mixed micelle).

Figure 5 Schematic illustration of Anticancer drug-loaded sterically stabilized micelle containing VIP peptide.

Figure 6 Schematic illustration of Anticancer drug-loaded sterically stabilized micelle containing SP94 peptide.

Figure 7 Schematic illustration of Anticancer drug-loaded sterically stabilized micelle containing VIP and H5WYG peptides.

Figure 8 Schematic illustration of Anticancer drug-loaded sterically stabilized micelle containing SP94 and H5WYG peptides.

Detailed explanation of the invention

The primary objective of the invention is, a cancer obliterator that contains anticancer drug enriched by H5WYG peptides containing several histidine amino acids with a property escaping from endosomes; and solves the problem of the vasoactive intestinal peptides and drug-loaded nano-particles taken into the cell (which play a role in endocytose process and specifically bind to the receptors on cancer cell membranes) not being released into the cytosol by being held by endosomes. It is a product that will be produced for the first time in sterically stabilized micelles group and one of its most important advantages is that is does not cause the side effects that occur with the other anti-cancer drugs.

The chemical structure of the materials involved in cancer obliterator which was the topic of our application and the structure of the developed derivatives of the obliterator have been clarified below;

Amphiphilic Compounds

They are the chemical compounds including both hydrophilic and hydrophobic structures. Amphiphilic phospholipids and polymer bound phospholipid compounds were used in this current invention.

Phospholipids

Phospholipids were utilized in this invention because of their biocompatibility, biodegradable characteristics and due to the ability to create anticancer drug loaded nanomicelle with targeting features. Phospholipids used in the current invention have different properties based on polar head group, non-polar chain length and the position of functional groups in the chain (DOPE, DOPC, DPPC etc.).

DOPC: 1 ,2-dioleoyl-sn-glycero-3-phosphocholine

DOPE: 1 ,2-dioleoyl-sn-glycero-3-phosphoethanolamin

DPPC:1 ,2-dipalmitoyl-sn-glycero-3-phosphocholine

Polymer bound phospholipid compounds

Polymer bound phospholipid compounds were used in this invention to increase their hydrophilic and escaping characteristic from immune cells in prepared nanomicelles. Poly(ethylene glycol) (PEG) bound phospholipid structures used in nanomicelle preparation have differences based on PEG chain length (n=500-1000) (DSPE-PEG2000, DSPE-PEG3400 etc.).

• DSPE-PEG2000:

1 ,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000 Vasoactive Intestinal Peptide (VIP) binds covalently DSPE-PEG-NHS polymer and the amphiphilic polymer is obtained. This polymer is in the structure of drug-loaded nano micelle, binds the cells that include VIP receptor, it provides the nano-micelle insures into the cell by active targeting. Nano-micelle structure that includes VIP and has the property of anticancer drug loaded active targeting is shown in the Figure 5.

Amino acid sequence of VIP peptide is shown below.

H-His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Thr-Arg-Leu-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr- Leu-Asn-Ser-lle-Leu-Asn-NH2

• Anticancer drug loaded SSM (Sterically stabilized micelle) and SSMM (Sterically stabilized mixed micelle)

The endothelial permeability increases when the junctions disappear between endothelial cell lines of tumor blood vessels. Therefore, drug loaded nanomicelles reach to the target region by passive targeting. It has the structure of drug loaded nano micelle occurring by the combination of phospholipids and/or polymer bound phospholipids with passive targeting features (Figure 3- 8).

• SP94GGC peptide:

Amphiphilic polymer is obtained by binding covalently with DSPE-PEG-MALEIMIDE polymer. This polymer is present in the structure of drug loaded nanomicelle, so it provides nanomicelle taken into the cell by active targeting binding to the receptors in HCC cells. The nanomicelle structure including SP94GGC with the property of anticancer drug loaded active targeting is shown in the Figure 6.

• Amino acid sequence of SP94GGC peptide is shown below.

^■ H-Ser-Phe-Ser-lle-lle-His-Thr-Pro-lle-Leu-Pro-Leu-Gly-Gly-Cys-COOH · Histidine aminoacid-rich H5WYGGGC peptide:

Amphiphilic polymer is obtained by binding covalently with DSPE-PEG-MALEIMIDE. This polymer is present in drug loaded nanomicelle structure and so it provides nanomicelle, which is taken into the cell by active targeting, to escape from endosomes. The nanomicelle structure including H5WYGGGC with the characteristic of anticancer drug loaded active targeting and endosomal escape is shown in the Figure 7,8.

• Amino acid sequence of H5WYGGGC peptide is shown below.

^■ Gly-Leu-Phe-His-Ala-lle-Ala-His-Phe-lle-His-Gly-Gly-Trp-His-Gly-Leu -lle- His-Gly-Trp-Thy-Gly-Gly-Gly-Cys-COOH

Anticancer drugs used in the samples were classified based on solvents used in the preparation step. Anticancer drug-loaded nanomicelles were generated with four different solvents (Methanol, ethanol, DMSO, water).

Methanol-soluble anticancer drugs; Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulphate, Bleomycin sulphate, Vindesine, Etoposide, Ibrutinib

DMSO (Dimethil - sulfoxid)-soluble anticancer drugs: Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib

• Ethanol-soluble anticancer drugs; Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib

^• Water-soluble anticancer drugs; Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate

SAMPLE 1

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulphate, Bleomycin sulphate, Vindesine, Etoposide, Ibrutinib) loaded nanomicel!e preparation:

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing methanol. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas till the methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours.

SAMPLE 2

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded nanomicelle preparation

DSPE- PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing DMSO. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until the DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours.

SAMPLE 3

Anticancer drug (Epothilone D,Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded nanomicelle preparation

DSPE- PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing ethanol. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours.

SAMPLE 4

Anticancer drug (Enzalutamide, Eriotinib Hydrochloride, Etoposide phosphate) loaded nanomicelle preparation DSPE- PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing water. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25°C in dark for 2 hours.

SAMPLE 5

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulfate, Bleomycin sulphate, Vindesine, Etoposide, Ibrutinib) loaded SSMM preparation

DOPC, DSPE- PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 3 vials containing methanol. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded SSMM generation at 25 °C in dark for 2 hours.

SAMPLE 6

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSMM preparation

DOPC, DSPE- PEG₂₀₀₀ and anticancer drug were vortexed untill they were completely dissolved in 3 vials containing DMSO. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded SSMM generation at 25 °C in dark for 2 hours. SAMPLE 7

Anticancer drug (Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSMM preparation

DOPC, DSPE- PEG₂₀₀₀ and anticancer drug were vortexed untill they were completely dissolved in 3 vials containing ethanol. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until the ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded SSMM generation at 25 °C in dark for 2 hours.

SAMPLE 8

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSMM preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 3 vials containing water. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded SSMM generation at 25 °C in dark for 2 hours.

SAMPLE 9

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulfate, Bleomycin sulfate, Vindesine, Etoposide, Ibrutinib) loaded SSMM-VIP preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing methanol. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nano micelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 10

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSMM-VIP preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing DMSO. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 11

Anticancer drug (Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSMM-VIP preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing ethanol. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v). SAMPLE 12

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSMM-VIP preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing water. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE- PEG3400-VIP and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 13

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulfate, Bleomycin sulfate, Vindesine, Etoposide, Ibrutinib) loaded SSMM-SM94 preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing methanol. Equal volumes were taken from DSPE-PEG_20-o and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94 and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 14

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSMM-SM94 preparation

DSPE-PEG2₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing DMSO. Equal volumes were taken from DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94 and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 15

Anticancer drug (Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSM-SP94 preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing ethanol. Equal volumes were taken from DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94 and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 16

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSMM-SM94 preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed till they were completely dissolved in 2 vials containing water. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic waterbath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE- PEG₃₄₀₀-SP94 and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v). SAMPLE 17

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulphate, Bleomycin sulphate, Vindesine, Etoposide, Ibrutinib) loaded SSM-H5WYG preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing methanol. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 18

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSM-H5WYG preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing DMSO. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 19

Anticancer drug (Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSM-H5WYG preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing ethanol. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 20

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSM-H5WYG preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing water. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE- PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 21

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing methanol. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v). SAMPLE 22

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing DMSO. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 23

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing ethanol. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 24

Anticancer drug (Enzalutamide, Eriotinib Hydrochloride, Etoposide phosphate) loaded SSMM-VIP preparation DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing water. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 25

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulfate, Bleomycin sulfate, Vindesine, Etoposide, Ibrutinib) loaded SSMM-SP94 preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing methanol. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94 and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 26

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSMM-SP94 preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing DMSO. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film 5 000320

was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94 and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 27

Anticancer drug (Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSMM-SP94 preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing ethanol. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for generation of anticancer drug loaded nanomicelle at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94 and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 28

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSMM-SP94 preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing water. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until the water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94 and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v). SAMPLE 29

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulfate, Bleomycin sulfate, Vindesine, Etoposide, Ibrutinib) loaded SSMM-H5WYG preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing methanol. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated.

The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 30

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSMM-H5WYG preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing DMSO. Equal volumes were taken from DOPC, DSPE- PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-H5\/VYG and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 31

Anticancer drug (Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSMM-H5WYG preparation P T/TR2015/000320

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing ethanol. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-H5\/VYG and anticancer drug loaded nanomicelle as the rate of 1 :4 (v/v).

SAMPLE 32

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSMM-H5WYG preparation

SAMPLE 33

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulfate, Bleomycin sulfate, Vindesine, Etoposide, Ibrutinib) loaded SSM-VIP-H5WYG preparation DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing methanol. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE- PEG₃₄₀₀-VIP, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v). SAMPLE 34

Anticancer drug (Doxorubicin, Cisplatin, ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSM-VIP-H5WYG preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing DMSO. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE- PEG₃₄₀₀-VIP, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1.1 :4 (v/v/v).

SAMPLE 35

Anticancer drug (Epothilone D.Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSM-VIP-H5WYG preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing ethanol. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE- PEG₃₄₀₀-VIP, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v).

SAMPLE 36

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSM-VIP-H5WYG preparation DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing water. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE- PEG3₄00-VIP, DSPE-PEG₃₄₀₀-H5VWG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v).

SAMPLE 37

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulfate, Bleomycin sulfate, Vindesine, Etoposide, Ibrutinib) loaded SSM-SP94-H5WYG preparation

DSPE-PEG₂₀₀0 and anticancer drug were vortexed until they were completely dissolved in 2 vials containing methanol. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE- PEG₃₄₀₀- SP94, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v).

SAMPLE 38

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSM-SP94-H5WYG preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing DMSO. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v).

SAMPLE 39

Anticancer drug (Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSM-SP94-H5WYG preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing ethanol. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v).

SAMPLE 40

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSM-SP94-H5WYG preparation

DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 2 vials containing water. Equal volumes were taken from DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE- PEG₃₄₀₀-SP94, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v). SAMPLE 41

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulfate, Bleomycin sulfate, Vindesine, Etoposide, Ibrutinib) loaded SSMM-VIP-H5WYG preparation

DOPC, DSPE-PEG2₀00 and anticancer drug were vortexed until they were completely dissolved in 3 vials containing methanol. Equal volumes were taken from DOPC, DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelie generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelie as the rate of 1 :1 :4 (v/v/v).

SAMPLE 42

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSMM-VIP-H5WYG preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing DMSO. Equal volumes were taken from DOPC, DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelie generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelie as the rate of 1 :1 :4 (v/v/v). SAMPLE 43

Anticancer drug (Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSMM-VIP-H5WYG preparation DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing ethanol. Equal volumes were taken from DOPC, DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v).

SAMPLE 44

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSMM-VIP-H5WYG preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing water. Equal volumes were taken from DOPC, DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-VIP, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v).

SAMPLE 45

Anticancer drug (Curcumine, Thymoquinone, Paclitaxel, Rapamycin, Vincristine sulfate, Bleomycin sulfate, Vindesine, Etoposide, Ibrutinib) loaded SSMM-SP94-H5WYG preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing methanol. Equal volumes were taken from DOPC, DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until methanol completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v).

SAMPLE 46

Anticancer drug (Doxorubicin, Cisplatin, Ixabepilone, Docetaxel, Amphotericin B, Carboplatin, Oxaliplatin, Methotrexate, 5-Fluorourasil, Gemcitabine, Vinblastine sulfate, Vinorelbine tartrate, Teniposide, Belinostat, Pomalidomide, Lenalidomide, Regorafenib, Dabrafenib, Everolimus, 10-hydroxy camptothecin, Cabozantinib, Vismodegib, Axitinib) loaded SSMM-SP94-H5WYG preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing DMSO. Equal volumes were taken from DOPC, DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until DMSO completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94, DSPE-PEG₃₄Q_O-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v). SAMPLE 47

Anticancer drug (Epothilone D, Colchicine, Mercaptopurine, Cyclophosphamide, Ceritinib, Afatinib, Trametinib, Bosutinib, Ponatinib, Pazopanib, Sunitinib malate, Vandetanib, Crizotinib, Vemorafenib) loaded SSMM-SP94-H5WYG preparation

DOPC, DSPE-PEG₂₀₀₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing ethanol. Equal volumes were taken from DOPC, DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until ethanol completely evacuated.

The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v). SAMPLE 48

Anticancer drug (Enzalutamide, Erlotinib Hydrochloride, Etoposide phosphate) loaded SSMM-SP94-H5WYG preparation

DOPC, DSPE-PEG200₀ and anticancer drug were vortexed until they were completely dissolved in 3 vials containing water. Equal volumes were taken from DOPC, DSPE-PEG₂₀₀₀ and anticancer drug solutions, and it was put in a round bottom volumetric flask and vortexed. It was vacuumed through the rotary evaporator under argon gas until water completely evacuated. The dried film was rehydrated with PBS (pH 7.4) and waited in ultrasonic water bath until the film was dissolved completely. Round bottom volumetric flask was closed with argon gas for anticancer drug loaded nanomicelle generation at 25 °C in dark for 2 hours. Solution was obtained by adding DSPE-PEG₃₄₀₀-SP94, DSPE-PEG₃₄₀₀-H5WYG and anticancer drug loaded nanomicelle as the rate of 1 :1 :4 (v/v/v).

Claims

1. The invention has composition of thymoquinone, phospholipids and polymer bound phospholipid mixtures, DSPE-PEG-VIP and DSPE-PEG-H5WYG compounds, it has also the characteristic of cancer obliterator with the feature of escaping from endosomes, thus solving the problem of drug loaded nanoparticles, which are used in cancer treatment, not to release into the cytosol by being held by the endosomes.

2. It is the invention mentioned in Claim 1 , the ratio between lipid and lipid types, DSPE-PEG- VIP, DSPE-PEG-H5WYG, thymoquinone generating drug loaded nanomicelle is between 0.01 and 100 and its optimum value is 1.

3. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes curcumine instead of the mentioned thymoquinone.

4. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes paclitaxel instead of the mentioned thymoquinone.

5. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes doxorubicin instead of the mentioned thymoquinone.

6. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes cisplatin instead of the mentioned thymoquinone.

7. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes rapamycin instead of the mentioned thymoquinone.

8. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes ixabepilone instead of the mentioned thymoquinone.

9. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes epothilone instead of the mentioned thymoquinone.

10. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes colchicine instead of the mentioned thymoquinone.

11. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes docetaxel instead of the mentioned thymoquinone.

12. lt has the feature of cancer obliterator according to Claim 1 and 2, and it includes 10- hydroxy camptothecin instead of the mentioned thymoquinone.

13. lt has the feature of cancer obliterator according to Claim 1 and 2, and it includes

Amphotericin B instead of the mentioned thymoquinone.

14. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes

Carboplatin instead of the mentioned thymoquinone.

15. lt has the feature of cancer obliterator according to Claim 1 and 2, and it includes

Oxaliplatin instead of the mentioned thymoquinone.

16. lt has the feature of cancer obliterator according to Claim 1 and 2, and it includes methotrexate instead of the mentioned thymoquinone.

17. lt has the feature of cancer obliterator according to Claim 1 and 2, and it includes 5- fluorourasil instead of the mentioned thymoquinone.

18. lt has the feature of cancer obliterator according to Claim 1 and 2, and it includes mercaptopurine instead of the mentioned thymoquinone.

19. lt has the feature of cancer obliterator according to Claim 1 and 2, and it includes gemcitabine instead of the mentioned thymoquinone.

20. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes bleomycin instead of the mentioned thymoquinone.

21. lt has the feature of cancer obliterator according to Claim 1 and 2, and it includes vincristine sulfate instead of the mentioned thymoquinone.

22. lt has the feature of cancer obliterator according to Claim 1 and 2, and it includes vinblastine sulfate instead of the mentioned thymoquinone.

23. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes vindesine instead of the mentioned thymoquinone.

24. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes vinorelbine tartrate instead of the mentioned thymoquinone.

25. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes cyclophosphamide instead of the mentioned thymoquinone.

26. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes etoposide instead of the mentioned thymoquinone.

27. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes etoposide phosphate instead of the mentioned thymoquinone.

28. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes teniposide instead of the mentioned thymoquinone.

29. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes belinostat instead of the mentioned thymoquinone.

30. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes Ibrutinib instead of the mentioned thymoquinone.

31. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes ceritinib instead of the mentioned thymoquinone.

32. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes afatinib instead of the mentioned thymoquinone.

33. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes trametinib instead of the mentioned thymoquinone.

34. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes pomalidomide instead of the mentioned thymoquinone.

35. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes Lenalidomide instead of the mentioned thymoquinone.

36. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes regorafenib instead of the mentioned thymoquinone.

37. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes dabrafenib instead of the mentioned thymoquinone.

38. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes everolimus instead of the mentioned thymoquinone.

39. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes bosutinib instead of the mentioned thymoquinone.

40. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes cabozantinib instead of the mentioned thymoquinone.

41. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes vismodegib instead of the mentioned thymoquinone.

42. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes ponatinib instead of the mentioned thymoquinone.

43. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes axitinib instead of the mentioned thymoquinone.

44. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes erlotinib hydrochloride instead of the mentioned thymoquinone.

45. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes omacetaxine mepesuccinate instead of the mentioned thymoquinone.

46. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes pazopanib instead of the mentioned thymoquinone.

47. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes enzalutamide instead of the mentioned thymoquinone.

48. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes sunitinib malate instead of the mentioned thymoquinone.

49. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes Vandetanib instead of the mentioned thymoquinone.

50. It has the feature of cancer obliterator according to Claim 1 and 2, and it includes crizotinib instead of the mentioned thymoquinone.

51. It has the feature of cancer obliterator according to Claim 1 , and it includes DSPE-PEG-

SP94 instead of DSPE-PEG-VIP.

52. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes curcumine instead of the mentioned thymoquinone.

53. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes paclitaxel instead of the mentioned thymoquinone.

54. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes doxorubicin instead of the mentioned thymoquinone.

55. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes cisplatin instead of the mentioned thymoquinone.

56. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes rapamycin instead of the mentioned thymoquinone.

57. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes ixabepilone instead of the mentioned thymoquinone.

58. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes epothilone instead of the mentioned thymoquinone.

59. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes colchicine instead of the mentioned thymoquinone.

60. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes docetaxel instead of the mentioned thymoquinone.

61. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes 10- hydroxy camptothecin instead of the mentioned thymoquinone.

62. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes

Amphotericin B instead of the mentioned thymoquinone.

63. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes Carboplatin instead of the mentioned thymoquinone.

64. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes

Oxaliplatin instead of the mentioned thymoquinone.

65. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes methotrexate instead of the mentioned thymoquinone.

66. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes 5- fluorourasil instead of the mentioned thymoquinone.

67. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes mercaptopurine instead of the mentioned thymoquinone.

68. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes gemcitabine instead of the mentioned thymoquinone.

69. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes bleomycin instead of the mentioned thymoquinone.

70. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes vincristine sulfate instead of the mentioned thymoquinone.

71. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes vinblastine sulfate instead of the mentioned thymoquinone.

72. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes vindesine instead of the mentioned thymoquinone.

73. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes vinorelbine tartrate instead of the mentioned thymoquinone.

74. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes cyclophosphamide instead of the mentioned thymoquinone.

75. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes etoposide instead of the mentioned thymoquinone.

76. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes etoposide phosphate instead of the mentioned thymoquinone.

77. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes teniposide instead of the mentioned thymoquinone.

78. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes belinostat instead of the mentioned thymoquinone.

79. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes Ibrutinib instead of the mentioned thymoquinone.

80. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes ceritinib instead of the mentioned thymoquinone.

81. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes afatinib instead of the mentioned thymoquinone.

82. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes trametinib instead of the mentioned thymoquinone.

83. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes pomalidomide instead of the mentioned thymoquinone.

84. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes

Lenalidomide instead of the mentioned thymoquinone.

85. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes regorafenib instead of the mentioned thymoquinone.

86. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes dabrafenib instead of the mentioned thymoquinone.

87. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes everolimus instead of the mentioned thymoquinone.

88. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes bosutinib instead of the mentioned thymoquinone.

89. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes cabozantinib instead of the mentioned thymoquinone.

90. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes vismodegib instead of the mentioned thymoquinone.

91. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes ponatinib instead of the mentioned thymoquinone.

92. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes axitinib instead of the mentioned thymoquinone.

93. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes erlotinib hydrochloride instead of the mentioned thymoquinone.

94. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes omacetaxine mepesuccinate instead of the mentioned thymoquinone.

95. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes pazopanib instead of the mentioned thymoquinone.

96. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes enzalutamide instead of the mentioned thymoquinone.

97. It has the feature of cancer obliterator according to Claim 2 and 51 , and it includes sunitinib malate instead of the mentioned thymoquinone.

98. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes

Vandetanib instead of the mentioned thymoquinone.

99. lt has the feature of cancer obliterator according to Claim 2 and 51 , and it includes crizotinib instead of the mentioned thymoquinone.

100. It has the feature of cancer obliterator according to Claim 1 , the mentioned nanomicelle has the composition having a mixture of only thymoquinone, phospholipids and/or polymer- bound phospholipids.

101. It is the invention stated in the Claim 100, wherein ratio between mixtures of phospholipids and / or polymer bound phospholipids forming drug loaded nanomisei and Timokinon is 0.01 to 100, the optimum value is 1.

102. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes curcumine instead of the mentioned thymoquinone.

103. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes paclitaxel instead of the mentioned thymoquinone.

104. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes doxorubicin instead of the mentioned thymoquinone.

105. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes cisplatin instead of the mentioned thymoquinone.

106. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes rapamycin instead of the mentioned thymoquinone.

107. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes ixabepilone instead of the mentioned thymoquinone.

108. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes epothilone instead of the mentioned thymoquinone.

109. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes colchicine instead of the mentioned thymoquinone.

110. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes docetaxel instead of the mentioned thymoquinone.

111. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes 10-hydroxy camptothecin instead of the mentioned thymoquinone.

112. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes Amphotericin B instead of the mentioned thymoquinone.

113. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes Carboplatin instead of the mentioned thymoquinone.

114. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes Oxaliplatin instead of the mentioned thymoquinone.

115. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes methotrexate instead of the mentioned thymoquinone.

116. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes 5- fluorourasil instead of the mentioned thymoquinone.

117. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes mercaptopurine instead of the mentioned thymoquinone.

118. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes gemcitabine instead of the mentioned thymoquinone.

119. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes bleomycin instead of the mentioned thymoquinone.

120. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes vincristine sulfate instead of the mentioned thymoquinone.

121. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes vinblastine sulfate instead of the mentioned thymoquinone.

122. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes vindesine instead of the mentioned thymoquinone.

123. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes vinorelbine tartrate instead of the mentioned thymoquinone.

124. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes cyclophosphamide instead of the mentioned thymoquinone.

125. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes etoposide instead of the mentioned thymoquinone.

126. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes etoposide phosphate instead of the mentioned thymoquinone.

127. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes teniposide instead of the mentioned thymoquinone.

128. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes belinostat instead of the mentioned thymoquinone.

129. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes Ibrutinib instead of the mentioned thymoquinone.

130. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes ceritinib instead of the mentioned thymoquinone.

131. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes afatinib instead of the mentioned thymoquinone.

132. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes trametinib instead of the mentioned thymoquinone.

133. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes pomalidomide instead of the mentioned thymoquinone.

134. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes Lenalidomide instead of the mentioned thymoquinone.

135. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes regorafenib instead of the mentioned thymoquinone.

136. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes dabrafenib instead of the mentioned thymoquinone.

137. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes everolimus instead of the mentioned thymoquinone.

138. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes bosutinib instead of the mentioned thymoquinone.

139. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes cabozantinib instead of the mentioned thymoquinone.

140. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes vismodegib instead of the mentioned thymoquinone.

141. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes ponatinib instead of the mentioned thymoquinone.

142. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes axitinib instead of the mentioned thymoquinone.

143. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes erlotinib instead of the mentioned thymoquinone.

144. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes omacetaxine mepesuccinate instead of the mentioned thymoquinone.

145. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes pazopanib instead of the mentioned thymoquinone.

146. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes enzalutamide instead of the mentioned thymoquinone.

147. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes sunitinib instead of the mentioned thymoquinone.

148. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes Vandetanib instead of the mentioned thymoquinone.

149. It has the feature of cancer obliterator according to Claim 100 and 101 , and it includes crizotinib instead of the mentioned thymoquinone.