CN109528686A

CN109528686A - Utilize the polymer-lipid mixing nano particle of microring array and the capecitabine of capecitabine amphipathic characteristic

Info

Publication number: CN109528686A
Application number: CN201710864852.9A
Authority: CN
Inventors: 程仲毅
Original assignee: PTM Biolabs Inc
Current assignee: PTM Biolabs Inc
Priority date: 2017-09-22
Filing date: 2017-09-22
Publication date: 2019-03-29

Abstract

The present invention includes composition and the method for preparing composition, the composition includes nano particle, the nano particle includes phosphatide core and at least one layer of one or more polymer on phosphatide core surface, and the phosphatide core includes one or more lipids and one or more active constituents；More particularly it relates to purposes of the capecitabine in the lipid-polymer nanoparticle formulations of such pharmaceutical properties for optimizing capecitabine (the fluoro- cytidine of N4- pentyloxy carbonyl -5- deoxidation -5-, CAP) in treating cancer.

Description

Utilize the polymer-lipid of microring array and the capecitabine of capecitabine amphipathic characteristic Mix nano particle

Inventive technique field

It is one or more poly- that the present invention relates generally at least one layer comprising phosphatide core and on the surface of the phosphatide core The nano particle of object is closed, the phosphatide core includes one or more lipids and one or more active constituents；More specifically, this hair It is bright to be related to such for mitigating the rouge of the side effect of capecitabine (the fluoro- cytidine of N4- pentyloxy carbonyl -5- deoxidation -5-, CAP) Purposes of the capecitabine in treating cancer in matter-polymer nanoparticle preparation.

Background of invention

It does not limit the scope of the invention, its background is described as about active medicine, more specifically capecitabine (N4- The fluoro- cytidine of pentyloxy carbonyl -5- deoxidation -5-, CAP) or its metabolin delivering.Although many United States Patent (USP)s provide cancer Learn the nanoparticle formulations of therapy medicine, but they are not with the lipid of capecitabine or Polymeric delivery, specifically card is trained The mixing delivering of the polymer-lipid of his shore or its metabolin is directly related.

Summary of the invention

Capecitabine (the fluoro- cytidine of N4- pentyloxy carbonyl -5- deoxidation -5-, CAP) is a kind of prodrug generally used, quilt For treating the multiclass cancer [bibliography 1-5] including colorectal cancer.In target tissue, CAP will be by enzyme from 5 '- '-Deoxy-5-fluorouridine is converted into 5 FU 5 fluorouracil, is a kind of active metabolite.Although CAP is in treatment such as colorectal cancer Kinds cancer in effectively, but half-life period is shorter.Drug is effectively removed from body in 0.5-1.0 hours, it is therefore desirable to High dose (150mg/m², twice daily).The large dosage of active component and/or peak value exposure can lead to more side effects, Such as Nausea and vomiting, dermatitis, bone marrow suppression, cardiac toxic and diarrhea.Therefore, sustained release and targeting particular cancers site Delivery system may improve treatment window.Nanoparticle formulations provide such possibility.We disclose preparations herein The method of stable capecitabine nano particle, the capecitabine nano particle control good, diameter particle diameter and surface Property is consistent with design.

The present invention includes composition and the method for preparing composition, the composition include containing one or more lipids and Phosphatide core including at least capecitabine or one or more active constituents of its metabolin and on the surface of phosphatide core At least one layer of one or more polymer.One or more lipids include at least one below: bis- capryl-sn- of 1,2- Glyceryl -3- phosphocholine (DDPC), 1,2- dilauroyl-sn- glyceryl -3- phosphoethanolamine (DLPE), two nutmegs Phosphatidyl choline (DMPC), dimyristoylphosphatidylglycerol (DMPG), bis- myristoyl-sn- glyceryl of 1,2-- 3- phosphoethanolamine (DMPE-PEG), 1- palmityl -2- myristoyl-sn- glyceryl -3- phosphocholine (PMPC), 1, Bis- palmityl-sn- glyceryl -3- phosphoric acid-(1 '-rac- glycerol) (DPPG) of 2-, bis- palmityl-sn- glyceryl -3- of 1,2- Phosphocholine (DPPC), bis- palmityl-sn- glyceryl -3- phosphoethanolamine (DPPE-PEG) of 1,2-, bis- palmityl of 1,2- Base-sn- glyceryl -3- phosphate (sodium salt) (DPPA), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphate choline (POPC), 1- palmityl -2- stearyl-sn- glyceryl -3- phosphocholine (PSOC), 1- stearyl -2- palmityl Base-sn- glyceryl -3- phosphocholine (SPPC), 1,2- distearyl acyl group-sn- glyceryl -3- phosphoethanolamine-N- [methoxyl group (polyethylene glycol) -2000] (DEPE-PEG), 1,2- distearyl acyl group-sn- glyceryl -3- phosphoethanolamine-N- [dibenzo ring Octyl (polyethylene glycol) -2000] (DSPE-PEG), L- α-phosphatidyl choline (L- α-PC), bis- sub-oleoyl-sn- glycerol of 1,2- Base -3- phosphocholine (DLPC), 1,2- dioleoyl-sn- glyceryl -3- phosphoric acid-(1 '-rac- glycerol) (DOPG), 1,2- bis- Stearyl-sn- glyceryl -3- phosphoric acid-(1 '-rac- glycerol) (DSPG), 1,2- distearyl acyl group -3- Trimethylammonium-Propane (DSTAP), 1,2- dioleyl-3-trimethylammonium-propane (DOTAP), 1,2- dioleoyl-sn- glyceryl -3- phosphate (DOPA), 1,2- dioleoyl-sn- glyceryl -3- phosphoethanolamine (DOPE), 1- stearyl -2- oleoyl-sn- glycerol Base -3- phosphocholine (SOPC), 1,2- dioleoyl-sn- glyceryl -3- phosphocholine (DOPC).In some aspects, Yi Zhonghuo A variety of active ingredients includes capecitabine (the fluoro- cytidine of N4- pentyloxy carbonyl -5- deoxidation -5-, CAP) or its metabolin.Activity at Dividing may include at least one anticancer drug；And/or selected from anticancer drug, antibiotic, antivirotic, antifungal agent, pest repellant, Nutrient, small molecule, at least one of siRNA, antioxidant and antibody.In some aspects, composition has high biology benefit Expenditure.In some aspects, active constituent may include traditional radioactive isotope.One or more active constituents include that water is insoluble Property dyestuff；And/or the metal nanoparticle to the contrast agent as MRI；And/or it is selected from Nile red, iron and platinum.In certain sides Face, one or more polymer include at least one below: poly- (lactic-co-glycolic acid) (PLGA) or its Pegylation Form PEG-PLGA, polylactic acid (PLA) or its PEGylated forms PEG-PLA, polyglycolic acid (PGA) or its Pegylation Form PEG-PGA, poly-L-lactide -co- 6-caprolactone (PLCL) or its PEGylated forms PEG-PLCL, hyaluronic acid (HA) or its PEGylated forms PEG-HA, poly- (- L-lysine) (PLL) or its PEGylated forms PEG-PLL, poly- third Olefin(e) acid (PAA) or its PEGylated forms PEG-PAA, polyphosphate (polyP), poly- (acrylic acid-co-maleic acid), poly- (fourth Succinate adipate), poly- (alkylcyanoacrylate) (PAC) or its PEGylated forms PEG-PAC.In many aspects, It is sweet that composition may include bis- palmityl-sn- glyceryl -3- phosphocholine (DPPC) of 1,2-, bis- palmityl-sn- of 1,2- Oil base -3- phosphoethanolamine (DPPE-PEG), L- α-phosphatidyl choline (L- α-PC), bis- sub-oleoyl-sn- glyceryl of 1,2- - 3- phosphocholine (DLPC) or 1,2- dioleyl-3-trimethylammonium-propane (DOTAP).Lipid and pegylated lipids Most common molar ratio is 100:0.01 to 50:50.Saturation lipid and unsaturated lipids molar ratio be usually 100:0.01 extremely 25:75.In some aspects, composition may include at least one targeting agent, wherein targeting agent with making nanoparticulate selective target To illing tissue/cell, thus minimize whole-body dose；And/or wherein targeting agent includes that can identify resisting for targeting antigen Body or its function fragment；And/or selected from antibody, small molecule, peptide, carbohydrate, siRNA, protein, nucleic acid, aptamer, the Two nano particles, cell factor, chemotactic factor (CF), lymphokine, receptor, lipid, agglutinin, ferrous metal, magnetic-particle, connection Son, isotope and their combination.In some aspects, nano particle has the diameter of 10nm to 200nm.Activity can be increased The bioavilability of ingredient mitigates side effect, and active constituent can be discharged in a continuous manner.

The present invention includes the embodiment for being used to form the method for nano particle comprising by by one or more phosphorus Rouge, one or more polymer, one or more solvents and the combination of at least one of capecitabine and its metabolin are to form Organic phase；Lipid water phase is formed by combining one or more targeting agents with water；Revolve organic phase and water phase in multiple entry To generate suspension thus the self assembly of micella occurs for mixing in the mixer of whirlpool；Spray drying or freeze-drying suspension simultaneously return Receive organic solvent；And mix the solution with one or more polymer with micella, layer-by-layer polymer deposits thus occur, To forming nano particle, and wherein when providing to object, nano particle provides continuing for capecitabine or its metabolin Release.

One or more polymer may include poly- (lactic-co-glycolic acid) (PLGA) or its PEGylated forms PEG-PLGA, polylactic acid (PLA) or its PEGylated forms PEG-PLA, polyglycolic acid (PGA) or its PEGylated forms PEG-PGA, poly-L-lactide -co- 6-caprolactone (PLCL) or its PEGylated forms PEG-PLCL, hyaluronic acid (HA), Polyacrylic acid (PAA) or its PEGylated forms PEG-PAA, polyphosphate (polyP), poly- (acrylic acid-co-maleic acid), Poly- (succinic acid-butanediol ester), poly- (alkylcyanoacrylate) (PAC) or its PEGylated forms PEG-PAC.

In many aspects, at least one solvent is selected from ethyl alcohol, methanol, tetrahydrofuran, acetonitrile, acetone, the tert-butyl alcohol, dimethyl Formamide and hexafluoroisopropanol.One or more active constituents include capecitabine or its metabolin；And/or at least one anticancer Drug；And/or traditional radioactive isotope；And/or at least one selected from fluorescent dye, quantum dot, iron, silver, gold and platinum is living Property ingredient.

In some aspects, one or more lipids may include 1,2-, bis- palmityl-sn- glyceryl -3- phosphocholine (DPPC), bis- palmityl-sn- glyceryl -3- phosphoethanolamine (DPPE-PEG) of 1,2-, L- α-phosphatidyl choline (L- α - PC), bis- sub-oleoyl-sn- glyceryl -3- phosphocholine (DLPC) of 1,2- or 1,2- dioleyl-3-trimethylammonium-propane (DOTAP)；And/or at least one lipid selected from the following: 1,2- didecyl docosahexaenoyl-sn-glycero -3- phosphocholine (DDPC), 1,2- dilauroyl-sn- glyceryl -3- phosphoethanolamine (DLPE), L-Dimyristoylphosphatidylcholine (DMPC), two meat Cardamom acyl group phosphatidyl glycerol (DMPG), bis- myristoyl-sn- glyceryl -3- phosphoethanolamine (DMPE-PEG) of 1,2-, 1- Palmityl -2- myristoyl-sn- glyceryl -3- phosphocholine (PMPC), bis- palmityl-sn- glyceryl -3- of 1,2- Phosphoric acid-(1 '-rac- glycerol) (DPPG), bis- palmityl-sn- glyceryl -3- phosphocholine (DPPC) of 1,2-, bis- palm of 1,2- Docosahexaenoyl-sn-glycero -3- phosphoethanolamine (DPPE-PEG), bis- palmityl-sn- glyceryl -3- phosphate (sodium salt) of 1,2- (DPPA), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphate choline (POPC), 1- palmityl -2- stearyl - Sn- glyceryl -3- phosphocholine (PSOC), 1- stearyl -2- palmityl-sn- glyceryl -3- phosphocholine (SPPC), 1,2- distearyl acyl group-sn- glyceryl -3- phosphoethanolamine-N- [methoxyl group (polyethylene glycol) -2000] (DEPE-PEG), 1, 2- distearyl acyl group-sn- glyceryl -3- phosphoethanolamine-N- [dibenzo cyclooctyl (polyethylene glycol) -2000] (DSPE- PEG), L- α-phosphatidyl choline (L- α-PC), bis- sub-oleoyl-sn- glyceryl -3- phosphocholine (DLPC) of 1,2-, 1,2- bis- Oleoyl-sn- glyceryl -3- phosphoric acid-(1 '-rac- glycerol) (DOPG), 1,2- distearyl acyl group-sn- glyceryl -3- phosphoric acid - (1 '-rac- glycerol) (DSPG), 1,2- distearyl acyl group -3- Trimethylammonium-Propane (DSTAP), 1,2- dioleoyl -3- front three Base ammonium-propane (DOTAP), 1,2- dioleoyl-sn- glyceryl -3- phosphate (DOPA), 1,2- dioleoyl-sn- glycerol Base -3- phosphoethanolamine (DOPE), 1- stearyl -2- oleoyl-sn- glyceryl -3- phosphocholine (SOPC), bis- oil of 1,2- Docosahexaenoyl-sn-glycero -3- phosphocholine (DOPC).

In some aspects, organic phase is mixed with lipid water phase including violent micro- mixed in multiple entry vortex mixer It closes；And/or mix organic phase with lipid water phase including being vortexed；And/or mixing organic phase with lipid water phase further includes ultrasound Processing.In some aspects, method further includes organic solvent removal；And/or dialysis；And/or freezing nanometer particle；And/or freeze-drying Nano particle；And/or spray-dried granules；And/or targeting agent is connect with nano particle；And/or at least one targeting of connection Agent, wherein targeting agent targets illing tissue/cell with making nanoparticulate selective, thus minimizes whole-body dose；And/or it will At least one targeting agent is connect with nano particle, and wherein targeting agent includes the antibody or its functional sheet that can identify targeting antigen Section.In some aspects, nano particle has the diameter of 10nm to 200nm.

The present invention includes the embodiment of the pharmaceutical agent comprising the nano particle for drug delivery, the nano particle The one or more polymer of at least one layer comprising lipid, active constituent and encapsulation lipid and active constituent.The present invention includes using In the embodiment for treating the doubtful patient with disease comprising application nano particle, wherein nano particle includes phosphatide core With the one or more polymer of at least one layer on the surface of phosphatide core, the phosphatide core include one or more phosphatide and At least one capecitabine and its active metabolite.In some aspects, application nano particle includes passing through intramuscular, subcutaneous, blood vessel It is interior or it is intravenous application and apply nano particle.Disease can be selected from tumour, nervous system and disease of metabolism；And/or choosing From parkinsons disease, Alzheimer's disease, multiple sclerosis, ALS, sequelae (sequel), behavior and cognitive disorder, certainly Close disease spectrum, depression and tumor disease；And/or cancer.In some aspects, active constituent is discharged in a continuous manner.

In a further embodiment, the method for treating the doubtful object with cancer includes: that identification is doubtful with cancer Patient；And at least one capecitabine and its metabolin for being enough to mitigate the amount of the cancer in object are provided to object, At least one of capecitabine and its metabolin be present in comprising one or more phosphatide and at least one capecitabine and its The phosphatide core of active metabolite；In at least one layer of one or more polymer on phosphatide core surface, wherein when to right When as providing, at least one of nano particle does not cause to include that Nausea and vomiting, dermatitis, bone marrow suppression, cardiac toxic and diarrhea exist One of interior side effect.In one aspect, cancer is breast cancer, colorectal cancer or cancer of pancreas.

Brief description

Feature and advantage for a more complete understanding of the present invention, referring now to detailed description of the invention and attached drawing, in which:

Fig. 1 is the figure for encapsulating the polymer-lipid nano particle of capecitabine.

Fig. 2 is the geometry of the center mixing portion of MIVM.

Fig. 3 is capecitabine molecular structure.

Fig. 4 is the CAP that is measured using low laser intensity (left side) and middle power intensity (right side) by DLS molten in DI water Solution is write music line.

Fig. 5 is the correlation function under different CAP concentration.(A) water, (B) 0.01mg/ml CAP are in DI water, (C) 0.1mg/ml CAP in DI water, (D) 0.5mg/ml CAP in DI water, (E) 2.5mg/ml CAP is in DI water, (F) 5mg/ Ml CAP in DI water, (G) 10mg/ml CAP in DI water, (H) 20mg/ml CAP is in DI water.

Fig. 6 is the structure of liposome-CAP micella.

Fig. 7 be micelle diameters distribution (A) CAP:DPPC:DPPE-PEG (30:40:30), (B) CAP:DPPE-PEG (30: 50) DLS data.Wherein x- axis is the diameter of nano particle, y- axis be to be formed nano-particle diameter quality of materials it is opposite Volume.

Fig. 8 is the CAP release from different micellar preparations.Wherein x- axis is the time, and y- axis is CAP from nano particle The percentage of cumulative release.

Fig. 9 is the diameter distribution of the micella of CAP, L- α-PC and DPPE-PEG.Wherein x- axis is the diameter of nano particle, y- Axis is the relative volume to form the quality of materials of nano-particle diameter.

Figure 10 is compared with pure CAP, and CAP is from the release in L- α-PC-DPPE-PEG micella.Wherein x- axis is time, y- Axis is the percentage of CAP cumulative release from nano particle.

Figure 11 is the distribution of DOTAP-CAP micelle diameters.Wherein x- axis is the diameter of nano particle, and y- axis is to form nanometer The relative volume of the quality of materials of grain diameter.

Figure 12 is CAP from the release profiles in DOTAP-CAP micella.Wherein x- axis is the time, and y- axis is CAP from nanometer The percentage of cumulative release in grain.

The structure of Figure 13 .polyP-DOTAP-CAP nano particle.

Figure 14, it is left: to be the diameter distribution of the DOTAP-CAP micella before adding polyP；It is right: to be after adding polyP The diameter of polyP-DPTAP-CAP nano particle is distributed.Wherein x- axis is the diameter of nano particle, and y- axis is to form nano particle The relative volume of the quality of materials of diameter.

Figure 15 is the zeta potential (left side) and diameter (right side) after the polyP-DOTAP-CAP nano particle of monitoring in five days.Wherein X- axis indicates the time point of measurement, and y- axis is the zeta potential of particle.

Figure 16 is compared with pure CAP micella, and CAP is from the release in polyP-DOTAP-CAP nano particle.Wherein x- axis is Time, y- axis are the percentage of CAP cumulative release from nano particle.

Figure 17 is the structure of PEG-PAA-DOTAP-CAP nano particle.

Figure 18 is the diameter distribution of PEG-PAA-DOTAP-CAP nano particle.Wherein x- axis is the diameter of nano particle, y- Axis is the relative volume to form the quality of materials of nano-particle diameter.

Figure 19 is the zeta potential (left side) of PEG-PAA-DOTAP-CAP nano particle in 5 days compared with polyP-DOTAP-CAP With diameter (right side).Wherein x- axis indicates the time point of measurement, and y- axis is the zeta potential of particle.

Figure 20 is the release profiles of DOTAP/CAP and PEG-PAA hybrid particles.Wherein x- axis is the time, y- axis be CAP from The percentage of cumulative release in nano particle.

Detailed description of the invention

When the formation and in use, the it should be understood that present invention of various embodiments of the present invention discussed further below Many suitable inventive concepts that can be embodied in a variety of specific contexts are provided.The specific embodiment being discussed herein is only to say It is bright to make and use concrete mode of the invention, it not delimit the scope of the invention.

In order to help to understand the present invention, multiple terms are defined below.Term defined herein has and such as has with the present invention The normally understood meaning of those of ordinary skill in the field of pass.Term for example "one", "an" and " described " be not intended to only Finger single entities, and the general category including that can be used to specific example illustrate.The term of this paper is for describing the present invention Specific embodiment, but in addition to as summarized in the claims, their use does not limit the present invention.

The treatment of cancer is limited by the side effect of anticancer drug.Chemotherapy is can for the limited of advanced cancer therapy In selection.However, the non-specific toxicity limit of these reagents of more and more drug resistance evidences and such as capecitabine Their therapeutic effect is made.It in order to overcome this problem, at cancer site to deliver drug is in right amount in vivo critically important. The new way of solution this problem is the drug delivery system by targeting, preferably delivers the medicament to cancer site.? In certain embodiments, using identification cancer cell and guide the miniature spherical particle (nano particle) of drug containing to the target of cancer cell To molecule (for example, antibody).

In certain embodiments, at least one targeting agent is connected to nano particle, and wherein targeting agent includes that can identify The antibody of targeting antigen or its function fragment.It can react heterologous/same with the amine of lipid and targeting moiety by being inserted into Source difunctional interval base connects targeting agent.

Capecitabine (the fluoro- cytidine of N4- pentyloxy carbonyl -5- deoxidation -5-, CAP) is for treating the general of colorectal cancer Time prodrug.In target tissue, it is a kind of activity that CAP, which will be converted to 5 FU 5 fluorouracil from 5 '-'-Deoxy-5-fluorouridines by enzyme, Metabolin.Although CAP has validity in treatment kinds cancer (i.e. colorectal cancer), CAP shows short drug Half-life period.Drug is removed from body in 0.5-1 hours, it is therefore desirable to high dose (150mg/m², twice daily).Large dosage It can lead to more side effects, such as Nausea and vomiting, dermatitis, bone marrow suppression, cardiac toxic and diarrhea.Therefore, sustained release with And the delivery system in targeting particular cancers site may improve treatment window.Nanoparticle formulations provide selection.It is of the invention public Having opened preparation has control good physical chemical property, such as the stable capecitabine nanometer of diameter diameter and surface nature The method of grain.Present inventors have recognized that the advantages of this lipid-PLGA mixing nanometer capecitabine, is high bioavilability, holds Continuous release, low internal removing simultaneously have the side effect mitigated.Since the pair of better bioavilability, sustained release and mitigation is made With the business potential of mixing nanometer capecitabine preparation is huge.

Nano particle structure

Capecitabine and phosphatide form micellar structure.Glue is wrapped up by electrostatic interaction using the polymer of opposite charges Beam.It if desired, can be by the polymer deposits of multilayer alternating charge on particle.Finally, outermost polyethylene glycol (PEG) mentions For spatial stability and long blood circulation time.

Method for forming particles

Liposome-capecitabine glue is generated by fast solvent exchange process using multiple entry vortex mixer (MIVM) first Beam is then spray-dried or is freeze-dried together with leucine and trehalose.Specifically, as shown in Figure 2, it is arranged MIVM.Micella suspension is freeze-dried 48 hours.By dry powder by be vigorously mixed or be ultrasonically treated be resuspended in it is water-soluble In liquid.The polymer of opposite charges is added to lipid-CAP micella.If plane SH wave on micella, finally uses poly- second Diolation diblock copolymer, to provide spatial stability.This method uses the amphipathic and controllable microring array of capecitabine.

Nanoparticle structure feature

Capecitabine it is amphipathic

The molecule displays of CAP are amphipathic out.As shown in Figure 3, capecitabine molecule has short carbon tail portion and hydrophily Head group (contains hydroxyl).Using dynamic light scattering (DLS) to measure solubility of the CAP in deionization (DI) water.Measurement CAP concentration is in 0.01mg/ml between 20mg/ml.Photon count rate keeps relatively low, until CAP concentration reaches several mg/ ml.Counting rate, which sharply increases, indicates that particle is formed in the solution.Experiment (Fig. 4) is repeated under low laser intensity and moderate strength. As a result significant difference is not present.In addition, by all CAP solution in left at room temperature over night and hereafter duplicate measurements.As a result it can weigh The scattered light intensity under the concentration of about a few mg/ml CAP is shown again to sharply increase.

DLS also is used to determine the diameter of particle, is about 2nm-30nm.Micellar structure is probably formed in the solution. However, particle diameter and correlation function fluctuate greatly under high CAP concentration, this, which shows CAP micella not, is very stable and balances It is quick (Fig. 5).

Liposome-CAP micella

Liposome is added to CAP to control the property of micella.Suitable liposome is listed in Table 1 below.Liposome-CAP The structure of micella is shown in Figure 6.

1. phosphatide of table and they the property of pH 7 list.The phosphatide of overstriking is for the phosphatide in this report.

General terms

Certain embodiments can be described as to the new formulation of the synthesis capecitabine in conjunction with PLGA and liposome Intravenous and/or subcutaneous administration.By such formulation design at providing the sustained release of the capecitabine as active constituent.It refers to Be polymer and being incorporated to for liposome components due to preparation and mitigate side effect.

In a further embodiment, composition can be used for treating tumor disease (cancer) and nervous system autoimmunity Degenerative disease (Parkinson's disease, Alzheimer's disease, multiple sclerosis, ALS, sequelae, behavior and cognitive disorder, from Close disease spectrum and depression).

In certain embodiments, composition of the invention is subjected to intramuscular, subcutaneous and/or intravascular application.

Certain embodiments include the capecitabine being encapsulated in liposome-PLGA encapsulating, and referred to as mixing Nano card trains him Shore preparation.

It should be understood that specific embodiments described here is shown by way of explanation but not as limit of the invention System.Without departing substantially from the scope of the present invention, main feature of the invention can be used in various embodiments.Ability The technical staff in domain will be appreciated that or just can determine using only routine experiment many equal similar shapes of specific procedure described herein Formula.Such equivalent form is deemed within the scope of the present invention and is covered by claim.

The all publications and patents application referred in the description shows the skill of those skilled in the art in the invention Art is horizontal.All publications and patents application is incorporated herein by reference, and degree is such as each independent publication or patent Shen It is please specifically and individually expressed as being incorporated by reference into.

When combine term " include (including) " in claim and/or specification in use, the word used "one" Or "an" can mean " one (kind) ", but its also be compliant with " one (kind) or multiple (kind) ", " at least one (kind) " and The meaning of " one (kind) or more than one (kind) ".The term "or" used in the claims for meaning "and/or", unless Clearly state and simply mean to optinal plan or optinal plan is mutually exclusive, but present disclosure support simply mean to optinal plan and The definition of "and/or".In entire application, term " about " is for indicating that numerical value includes for measuring the device of the value, method Change present in intrinsic error change or research object.

As used in the specification and claims, word "comprising", " having ", " comprising " or " containing " are inclusive Or it is open and be not excluded for other, unlisted element or method and step.

The term as used herein " or their combination " refers to all arrangements and combination that project is listed before the term. For example, " A, B, C or their combination " is intended to include at least one below: A, B, C, AB, AC, BC or ABC, and if suitable Sequence is critically important in specific context, then further includes BA, CA, CB, CBA, BCA, ACB, BAC or CAB.Continue the example, it is bright Really include the duplicate combination containing one or more projects or term, for example, BB, AAA, AB, BBC, AAABCCCC, CBBAAA, CABABB etc..It will be appreciated by those skilled in the art that the number of project or term in any combination there is usually no Limitation, unless in addition apparent from the context.In certain embodiments, the present invention can also include method and composition, Can also wherein use transition phrase " substantially by ... form " or " by ... form ".

According to present disclosure, it can prepare and implement disclosed herein and claimed all compositions and/or side Method, without excessive experiment.Although having described the compositions and methods of the invention in a manner of preferred embodiment, to this It, can be to combination described herein without departing substantially from design of the invention, spirit and scope for the technical staff of field Object and/or method and the method the step of in or in the sequence of step using variation be obvious.To this field skill Obvious all such similar alternatives and modifications are considered as sending out at as defined in the appended claims for art personnel In bright spirit, scope and spirit.

Multiple polymers and the method that particle is consequently formed are known in the field of drug delivery.Of the invention one In a little embodiments, the matrix of particle includes one or more polymer.Any polymer can be used according to the present invention.Polymerization Object can be natural polymer or non-natural (synthesis) polymer.Polymer can be homopolymer or comprising two or more list The copolymer of body.According to sequence, copolymer can be random, block, or including random and block order combination.In general, Polymer according to the present invention is organic polymer.

As used herein, " polymer " is endowed its common meaning as used in this field, that is, molecular structure includes one Kind or a variety of repetitive units (monomer) by being covalently keyed.Repetitive unit can be all the same, or in some cases Under, there can be the repetitive unit of more than one type to be present in polymer.In some cases, polymer is biological source , that is, biopolymer.The non-limiting example of biopolymer includes peptide or protein matter (that is, the polymerization of various amino acid Object) or such as DNA or RNA.In some cases, part in addition can also exist in polymer, such as described below Biological part retinal diseases.If the repetitive unit of more than one type is present in polymer, polymer is referred to as " copolymer ".It answers Understand, in any embodiment using polymer, the polymer used in some cases can be copolymer. The repetitive unit for forming copolymer can be arranged in any way.For example, repetitive unit can with random order, with alternately Sequence is arranged, or is arranged in " block " copolymer, that is, one or more areas comprising respectively containing the first repetitive unit Domain (for example, first block) and the one or more regions (for example, second block) for respectively containing the second repetitive unit etc..Block Copolymer can have the different blocks of two (diblock copolymers), three (triblock copolymer) or more.Of the invention Multiple embodiments are related to copolymer, describe in specific embodiments usually through two or more polymer one Two or more polymer (such as polymer described herein) for playing covalent bonding and being bonded to each other.Therefore, copolymer It may include first polymer and second polymer, they are combined together to form block copolymer, and wherein first polymer is The first block and second polymer of block copolymer are the second blocks of block copolymer.Certainly, ordinary skill Personnel should be understood that in some cases block copolymer can contain multiple polymer blocks, and should be understood that used herein " block copolymer " is not limited only to block copolymer only with single first block and single second block.For example, block is total Polymers may include comprising the first block of first polymer, the second block comprising second polymer and comprising third polymer Or third block of first polymer etc..In some cases, block copolymer can contain any number of first polymer The first block and second polymer the second block (and in some cases, third block, the 4th block etc.).In addition, It should be noted that in some instances, block copolymer can also be formed by other block copolymers.For example, the first block is total Polymers can be combined with other polymer (it can be homopolymer, biopolymer, block copolymer in addition etc.), with shape The block copolymer containing polymorphic type block of Cheng Xin, and/or combined with other parts (for example, non-polymeric part).Optionally, As described below, lipid connexon (for example, DSPE), which can be used, forms copolymer.

In one group of embodiment, polymer of the invention (for example, copolymer, for example, block copolymer) includes biology Compatible polymer, that is, when in the object of insertion or injection work, the polymer does not cause adverse reaction, such as nothing usually Significant inflammation and/or pass through immune system (for example, via t cell response) the acute cellular rejection polymer.Therefore, of the invention Nano particle can be " non-immunogenic ".The term as used herein " non-immunogenic " refers in its native state Source property growth factor, do not cause antibody, T- cell or reactive immunocyte circulation usually or only cause antibody, T- cell or The floor level of reactive immunocyte circulation, and it does not cause the immune response that itself is directed in individual usually.

Certainly, it should be recognized that " biocompatibility " is relative terms, and expectable immune response to a certain degree, even if For the polymer compatible with tissue height living also so.However, as used herein, " biocompatibility " refers at least Acute cellular rejection of a part of immune system to material, that is, the non-biocompatible material for being implanted into object causes being immunized in object Reaction, seriously prevents immune system to the repulsion of material from being suitably controlled, and be usually such that material enough The degree that must be removed from object.A simple experiment for measuring biocompatibility is that polymer is exposed to cell in vitro； Biocompatible polymer is such polymer, under intermediate concentration, for example, under the concentration of 50 microgram/10 cells, It usually will not result in significant cell death.For example, when being exposed to the cell of such as fibroblast or epithelial cell, biology Compatible polymer can cause the cell death less than about 20%, even if being swallowed by such cell or other modes absorb also such as This.The non-limiting example for the biocompatible polymer that can be used in various embodiments of the present invention includes poly- dioxanes Ketone (PDO), polyhydroxyalkanoatefrom, poly butyric ester, poly- (glycerol sebacate), polyglycolide, polylactide, PLGA, Polycaprolactone or copolymer or derivative comprising these polymer and/or other polymers.

In certain embodiments, biocompatible polymer is biodegradable, that is, in physiological environment, such as In vivo, polymer being capable of chemical degradation and/or biological degradation.For example, polymer can be when be exposed to water (for example, In object) when the polymer that spontaneously hydrolyzes, when being exposed to hot (for example, at a temperature of about 37 DEG C), polymer can degrade. According to the polymer or copolymer used, the degradation of polymer can occur at a different rate.For example, according to polymer, The half-life period (50% polymer is degraded into monomer and/or the time of other non-polymeric portions) of polymer can be approximate number It, several weeks, several months or several years.Such as by enzymatic activity or cell mechanism, in some cases, such as by being exposed to lysozyme (for example, having relatively low pH), polymer can carry out biological degradation.In some cases, polymer can be decomposed At monomer and/or other non-polymeric portions (for example, polylactide can be hydrolyzed to form lactic acid, polyglycolide can be into Row hydrolyzes to form glycolic etc.), in the case where not having significant toxic effect to cell, cell can be re-used or be removed The monomer and/or other non-polymeric portions.In some embodiments, polymer can be polyester comprising: include cream The copolymer of acid and glycolic acid units, such as poly- (lactic-co-glycolic acid) and poly(lactide-co-glycolide), herein It is referred to as " PLGA "；And the homopolymer comprising glycolic acid units, referred to herein as " PGA ", and the homopolymerization comprising lactic acid units Object, such as Poly-L-lactide, poly- D-ALPHA-Hydroxypropionic acid, poly- D, Pfansteihl, poly-L-lactide, poly- D- lactide and poly- D, L- third are handed over Ester, referred to herein, generally, as " PLA ".In some embodiments, Exemplary polyesters include, for example, polyhydroxy acid；Lactide and second The pegylated polymer and copolymer of lactide are (for example, Pegylation PLA, Pegylation PGA, Pegylation PLGA and their derivative.In some embodiments, polyester includes, for example, polyanhydride, poly- (ortho esters) polyethylene glycol Change poly- (ortho esters), poly- (caprolactone), Pegylation poly- (caprolactone), polylysine, Pegylation polylysine, gather (aziridine), Pegylation poly- (aziridine), poly- (L- lactide-co-L lysine), poly- (serine ester), poly- (4- Hydroxy-L-proline ester), poly- [a- (4- aminobutyl)-L glycolic] and their derivative.

In some embodiments, polymer can be PLGA.PLGA be bio-compatible and biodegradable lactic acid and The copolymer of glycolic, and various forms of PLGA are characterized in that lactic acid: the ratio of glycolic.Lactic acid can be L- cream Acid, D-ALPHA-Hydroxypropionic acid or D, Pfansteihl.The degradation rate of PLGA can be adjusted by changing the ratio of lactic acid-ethanol.? In some embodiments, PLGA ready for use is characterized in that lactic acid according to the present invention: the ratio of glycolic is about 85:15, about 75:25, about 60:40, about 50:50, about 40:60, about 25:75 or about 15:85.

In specific embodiments, by optimize nano particle polymer (for example, PLGA block copolymer or PLGA-PEG block copolymer) in lactic acid and glycolic acid monomers ratio, can optimize such as water imbibition, drug release (for example, " control release ") and polymer degradation kinetics nano particle parameter.In some embodiments, polymer can be one kind Or a variety of acrylate copolymers.In certain embodiments, acrylate copolymer includes, for example, acrylic acid and methacrylic acid Copolymer, methylmethacrylate copolymer, ethoxyethyl methacrylates copolymer, the copolymerization of methacrylic acid cyanaoethyl methacrylate Object, amino alkyl methacrylate copolymer, poly- (acrylic acid), poly- (methacrylic acid), methacrylic acid alkylamide are total Polymers, poly- (methyl methacrylate), poly- (methacrylic acid polyacrylamide, amino alkyl methacrylate copolymer, first Base glycidyl acrylate copolymer, polybutylcyanoacrylate and the combination comprising one or more aforementioned polymers.Third Olefin(e) acid polymer may include have the quaternary ammonium group of low content acrylate and methacrylate polymerize completely be total to Polymers.

In some embodiments, polymer can be cationic polymer.In general, cationic polymer can flocculate And/or the electronegative nucleic acid chains (such as DNA, RNA or their derivative) of protection.Amine-containing polymer, such as poly- (bad ammonia Acid) (Zauner et al., 1998, Adv.Drug Del.Rev., 30:97；And Kabanov et al., 1995, Bioconjugate Chem., 6:7), polyethyleneimine) (PEI；Boussif et al., 1995, Proc.Natl.Acad.ScL, USA, 1995,92: 7297) and poly- (amide amine) dendritic macromole (Kukowska-Latallo et al., 1996, Proc.Natl.Acad.ScL,USA,93:4897；Tang et al., 1996, Bioconjugate Chem., 7:703；And Haensler et al., 1993, Bioconjugate Chem., 4:372), it is positively charged at physiological ph, ion is formed with nucleic acid It is right, and the mediated transfection in multiple cell lines.

In some embodiments, polymer can be it is degradable with cationic side chain polyester (Putnam et al., 1999,Macromolecules,32:3658；Barrera et al., 1993 ,/.Am.Chem.Soc, 115:11010；Kwonef Et al., 19%9, Macromolecules, 22325Q-, Urn et al., 1999, J.Am.Chem.Soc, 121:5633；And Zhou et al., 1990, Macromolecules, 23:3399).The example of these polyester includes that poly- (L- lactide-co-L- relies ammonia Acid) (Barrera et al., 1993 ,/.Am.Chem.Soc, 115:11010), poly- (serine ester) (Zhou et al., 1990, Macromolecules, 23:3399), poly- (4-hydroxy-L-proline ester) (Putnam et al., 1999, Macromolecules, 32:3658；And Lim et al., 1999 ,/.Am.Chem.Soc, 121:5633).Prove that poly- (4-hydroxy-L-proline ester) is logical Cross electrostatic interaction flocculation Plasmid DNA, and mediate gene transfer (Putnam et al., 1999, Macromolecules, 32: 3658；And Lim et al., 1999 ,/.Am.Chem.Soc, 121:5633).These new polymer are than poly- (lysine) and PEI Toxicity is low, and they are degraded into nontoxic metabolin.Containing poly(ethylene glycol) repetitive unit polymer (for example, copolymer, For example, block copolymer) it is also known as " Pegylation " polymer.Due to the presence of poly(ethylene glycol) group, this Type of Collective Object can control inflammation and/or immunogenicity (that is, the ability for causing immune response) and/or reduce via reticuloendothelial system from The clearance rate of the circulatory system.In some cases, polymer can also be reduced using Pegylation and Division of Biological Science divides it Between charge interaction, for example, by generating hydrophilic layer on the surface of polymer, can protect polymer from Biological part retinal diseases interaction.In some cases, the addition of poly(ethylene glycol) repetitive unit can increase polymer (for example, Copolymer, for example, block copolymer) plasma half-life, for example, it is absorbed by reducing polymer by mononuclear phagocyte system, Transfection/the absorption efficiency for reducing cell simultaneously carries out.Those of ordinary skill in the art, which will be known, is used for pegylated polymer Methods and techniques, for example, by using EDC (l- ethyl -3- (3- dimethylaminopropyl) carbodiimide hydrochloride) and NHS (n-hydroxysuccinimide) is so that polymer is reacted with the PEG group blocked with amine, by ring-opening polymerisation technology (ROMP) etc..

In addition, certain embodiments of the present invention are related to the copolymer containing poly- (ester-ether), lead to for example, polymer has Cross the repetitive unit of ester bond (for example, R-C (O)-O-R ' key) and ehter bond (for example, R-O-R ' key) connection.Of the invention some In embodiment, biodegradable polymer, such as the hydrolysable polymeric containing carboxylic acid group, it can be with poly(ethylene glycol) weight Multiple unit is combined to form poly- (ester-ether).

In specific embodiments, optimize the molecular weight of nanoparticulate polymer of the invention for effectively treating such as The diseases such as cancer.For example, the molecular weight effects nano particle degradation rate of polymer is (especially when adjustment biological degradation polyalcohol When molecular weight), solubility, water imbibition and drug release kinetics (such as " controlled release ").As another example, adjustable The molecular weight of polymer makes nano particle (several small in carrying out biodegrade in the reasonable period in treated object Up to 1-2 weeks, 3-4 weeks, 5-6 weeks, 7-8 weeks etc.).In the specific embodiment party of the nano particle comprising PEG and the copolymer of PLGA In case, the molecular weight of PEG is 1,000-20,000, such as 5,000-20,000, such as 10,000-20,000, and PLGA Molecular weight is 5,000-100,000, such as 20,000-70,000, such as 20,000-50,000.

In other embodiment, the present invention provides the nano particles of amphipathic layer protection, and prepare the nanometer The method of particle, wherein by a kind of polymer (for example, PEG) of polymer substrate and will be with another polymer (for example, PLGA) The lipid binding of self assembly, so that the polymer of polymer substrate not instead of covalent bond, is combined by self assembly." from group Dress " refers to the process of the spontaneous assembling of higher order structure, and the component (for example, molecule) dependent on higher order structure is mutual naturally to draw Power.

It is typically based on diameter, shape, composition or chemical property and is occurred by the random motion of molecule and the formation of key. The lipid of self assembly for polymer is the component in addition to the amphipathic component of nano particle.

In certain embodiments, the polymer of nano particle can be with lipid, that is, except the two of nano particle of the invention Lipid binding except parent's property component.Polymer can be the PEG of such as lipid sealing end.The present invention also provides be used to form to have The method of the nano particle of the amphipathic protection of the PEG of lipid sealing end.For example, the method includes first provided with lipid reactant Polymer, to form polymer/lipid conjugates.Then, polymer/lipid conjugates are reacted with targeting moiety to prepare targeting Polymer/lipid conjugates that part combines；And keep polymer/lipid conjugates of ligand binding and second polymer (non- Functional group's fluidized polymer), amphipathic component and drug mixing；So that forming the nano particle of amphipathic layer protection.In certain realities It applies in scheme, first polymer PEG, so that forming the PEG of lipid sealing end.Then, the PEG of lipid sealing end can be with, for example, with PLGA is mixed to form nano particle.As described above, the lipid part of polymer can be used for other polymer self assembles, Contribute to form nano particle.For example, hydrophilic polymer can be with the lipid binding with hydrophobic polymer self assembly.

Liposome

In some embodiments, lipid is oil.In general, any oil known in the art can gather with for of the invention Object is closed to combine.In some embodiments, oil may include one or more fatty acid groups or their salt.In some implementations In scheme, fatty acid group may include digestible long-chain (for example, Cg-Cso), substituted or unsubstituted hydrocarbon.In some realities It applies in scheme, fatty acid group can be C₁₀-C₂₀Fatty acid or its salt.In some embodiments, fatty acid group can be C₁₅-C₂₀Fatty acid or its salt.In some embodiments, fatty acid can be unsaturated.In some embodiments, rouge Fat acid groups can be monounsaturated.In some embodiments, fatty acid group can be how unsaturated.In some realities It applies in scheme, the double bond of unsaturated fat acid groups can be cisoid conformation.In some embodiments, unsaturated fatty acid Double bond can be anti conformation.

In some embodiments, fatty acid group can be butyric acid, caproic acid, octanoic acid, capric acid, lauric acid, myristic acid, One of palmitinic acid, stearic acid, arachidic acid, behenic acid or lignoceric acid are a variety of.In some embodiments, rouge Fat acid groups can be palmitoleic acid, oleic acid, octadecenoic acid, linoleic acid, alpha-linolenic acid, gamma-linoleic acid, arachidonic acid, One of cis-9-20 carbon acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid or erucic acid are a variety of.

The property of these polymer and other polymers and the method for being used to prepare them are (ginsengs known in the art See, for example, United States Patent (USP) 6,123,727；5,804,178；5,770,417；5,736,372；5,716,404；6,095,148； 5,837,752；5,902,599；5,696,175；5,514,378；5,512,600；5,399,665；5,019,379；5,010, 167；4,806,621；4,638,045；With 4,946,929；Wang et al., 2001 ,/.Am.Chem.Soc, 123:9480；Lim Et al., 2001 ,/.Am.Chem.Soc, 123:2460；Langer,2000,Ace.Chem.Res.,33:94；Langer, 1999,/.Control.Release,62:7；And Uhrich et al., 1999, Chem.Rev., 99:3181).More generally, A variety of methods for synthesizing suitable polymer are described in the following: Concise Encyclopedia of Polymer Science and Polymeric Amines and Ammonium Salts, is write by Goethals, Pergamon Press,1980；Principles of Polymerization by Odian,John Wiley&Sons).

Pharmaceutical composition

As used herein, term " pharmaceutically acceptable carrier " means any kind of nontoxic, inert solid, semisolid Or liquid filler, diluent, encapsulating material or formulation auxiliary agents.Remington ' s Pharmaceutical Sciences. by Gennaro writes, and Mack Publishing, Easton, Pa., 1995 disclose the various carriers for compounding pharmaceutical composition And it is used to prepare the known technology of the carrier.The some examples that can be used as the material of pharmaceutically acceptable carrier include, but It is not limited to: sugar, such as lactose, dextrose and saccharose；Starch, such as cornstarch and potato starch；Cellulose and its derivates, Such as sodium carboxymethylcellulose, ethyl cellulose and cellulose ethanoate；Powdered tragacanth；Malt；Gelatin；Talcum；Excipient, Such as cupu oil and suppository wax；Oil, such as peanut oil, cottonseed oil；Safflower oil；Sesame oil；Olive oil；Corn oil and soya-bean oil；Two Alcohol, such as propylene glycol；Ester, such as ethyl oleate and ethyl laurate；Agar；Detergent, such as Tween 80；Buffer, such as Magnesium hydroxide and aluminium hydroxide；Alginic acid；Apirogen water；Isotonic saline solution；Ringer's solution；Ethyl alcohol；And phosphate buffer solution, with And other non-toxic compatible lubricants, such as lauryl sodium sulfate and magnesium stearate, and according to the judgement of formulator, Can also there are colorant, releasing agent, coating agent, sweetener, flavoring agent and aromatic, preservative and anti-oxidant in the composition Agent.If filtering or other final method for disinfection are infeasible, preparation can be aseptically prepared.

Pharmaceutical composition of the invention can be applied by any method known in the art to patient, the method includes Oral route and parenteral route.In certain embodiments, parenteral route is desired, because they avoid and are found in Digestive ferment contact in alimentary canal.According to such embodiment, composition of the invention can by injection (for example, it is intravenous, Subcutaneously or intramuscularly, intraperitoneal injection), per rectum, Via vagina, part (as passed through pulvis, creme, ointment or drops) or pass through Sucking (as by spraying) is administered.In specific embodiments, nano particle of the invention is for example defeated by vein It infuses or injects to the object systemic administration for thering is this to need.

According to known technology using suitable dispersing agent or wetting agent and suspending agent, injection preparation can be prepared, for example, Sterile injectable aqueous or oil-based suspension.Sterile injectable preparation can also be in the nontoxic acceptable diluent of parenteral Or sterile injectable solutions, suspension or lotion in solvent, for example, as the solution in 1,3-BDO.It can be used Acceptable medium and solvent be water, Ringer's solution, U.S.P. and isotonic sodium chlorrde solution.In addition, usually will be sterile Fixed oil is used as solvent or suspension media.For this purpose, any mild fixed oil, including synthesis can be used Monoglyceride or double glyceride.In addition, the fatty acid of such as oleic acid is used to prepare injection.In one embodiment, originally The conjugate of invention is suspended in the carrier fluid comprising 1% (w/v) sodium carboxymethylcellulose and 0.1% (v/v) Tween 80.Example Such as, it can be filtered by retaining the filter of bacterium, or (it is being used the bactericidal agent by mixing aseptic solid composite form Before can be dissolved or dispersed in sterile water or other sterile injectable mediums), so that injection preparation be sterilized.

Composition for rectum or vaginal application can be suppository, can by make conjugate of the invention with such as The suitable nonirritant excipient or carrier of cupu oil, polyethylene glycol or suppository wax mix to be prepared, and the suppository exists It is solid under environment temperature and is under body temperature liquid, and is therefore fused in rectum or vaginal canal and discharges conjugation of the present invention Object.Dosage form for part or the pharmaceutical composition of the invention of transdermal administration includes ointment, paste, creme, lotion, gel Agent, pulvis, solution, spray, inhalant or patch.According to possible needs, aseptically by conjugate of the invention It is mixed with pharmaceutically acceptable carrier and any desired preservative or buffer.Eye-drops preparations, auristilla and eye drops Also it is included within the scope of the invention.Ointment, paste, creme and gelling agent remove containing invention conjugate of the invention it Outside, can also contain: excipient, such as animal tallow and plant fat, oil, wax, paraffin, starch, bassora gum, cellulose derive Object, polyethylene glycol, silicone, bentonite, silicic acid, talcum and zinc oxide or their mixture.Transdermal skin patches have to be mentioned to body For the other advantage of the controllable delivery of compound.Such dosage form can be by the way that conjugate of the present invention to be dissolved or dispersed in suitably Medium in prepared.The flux that compound passes through skin can also be increased using absorption enhancer.Rate can pass through Rate controlling membranes are provided or are controlled by dispersing conjugate of the present invention in polymer substrate or gel.

Pulvis and spray can also contain excipient, such as lactose, cunning in addition to containing invention conjugate of the invention Stone, silicic acid, aluminium hydroxide, calcium silicates and polyamide powder or their mixture.In addition spray can contain commonly pushes away Into agent, such as chlorofluorocarbons.When being administered orally, nano particle of the invention can be packaged, but be not required.Various conjunctions Suitable package system is (" Microcapsules and Nanoparticles in Medicine and known in the art Pharmacy " is edited by Doubrow, M., CRC Press, Boca Raton, and 1992；Mathiowitz and Langer J.Control.Release 5:13,1987；Mathiowitz et al. .Reactive Polymers 6:275,1987； Mathiowitz et al. .J.Appl.Polymer Sci.35:755,1988；Langer Ace.Chem.Res.33:94,2000； Langer J.Control.Release 62:7,1999；Uhrich et al. .Chem.Rev.99:3181,1999；Zhou et al. .J.Control.Release 75:27,2001；And Hanes et al., Pharm.Biotechnol.6:389,1995).This hair Bright conjugate can be encapsulated in biodegradable polymer microballoon or liposome.It is used to prepare the day of biodegradable microspheres So includes carbohydrate such as alginates, cellulose, polyhydroxyalkanoatefrom, polyamide, gathers with the example of synthetic polymer Phosphonitrile, poly- propyl fumarate, polyethers, polyacetals, polybutylcyanoacrylate, Biodegradable polyurethane, polycarbonate, polyacids Acid anhydride, polyhydroxy acid, poly- (ortho esters) and other biological degradable polyester.The example that can be used for the lipid of liposome preparation includes phosphorus Acyl compound, for example, phosphatidyl glycerol, phosphatidyl choline, phosphatidylserine, phosphatidyl-ethanolamine, sphingolipid, cerebroside and Gangliosides.Pharmaceutical compositions for oral administration can be liquid or solid.Suitable for present invention combination is administered orally The liquid dosage form of object includes pharmaceutically acceptable lotion, microemulsion, solution, suspension, syrup and elixir.In addition to encapsulating or not sealing The conjugate of dress, liquid dosage form can also contain inert diluent commonly used in the art, for example, water or other solvents, solubilising Agent and emulsifier, such as ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, benzylalcohol, Ergol, propylene glycol, 1,3- fourth Glycol, dimethylformamide, oil (especially cottonseed oil, peanut oil, corn oil, embryo oil, olive oil, castor oil and sesame Oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan aliphatic ester and their mixture.

Other than inert diluent, Orally administered composition can also include adjuvant, wetting agent, emulsifier and suspending agent, sweet taste Agent, flavoring agent and aromatic.As used herein, term " adjuvant " refers to anyization of the non-specific regulator for immune response Close object.In certain embodiments, adjuvant challenge.Any adjuvant can be used according to the present invention.A large amount of adjuvant Compound is (Allison Dev.Biol.Stand.92:3-11,1998 known in the art；Unkeless et al. .Annu.Rev.Immunol.6:251-281,1998；And Phillips et al. .Vaccine10:151-158,1992).

Solid dosage forms for oral administration includes capsule, tablet, pill, powders and granules.In such solid dosage forms In, encapsulation or unencapsulated conjugate and at least one inert pharmaceutically acceptable excipient or carrier such as sodium citrate or Calcium monohydrogen phosphate and/or following substance are mixed: (a) filler or incremental agent, such as starch, lactose, sucrose, glucose, sweet Reveal pure and mild silicic acid, (b) adhesive, for example, carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and I Primary glue, (c) moisturizer such as glycerol, (d) disintegrating agent such as aga agar, calcium carbonate, potato or tapioca, alginic acid, certain silicon Hydrochlorate and sodium carbonate, (e) solution resistance solvent such as paraffin, (f) sorbefacient such as quaternary ammonium compound, (g) wetting agent, such as spermaceti Pure and mild glycerin monostearate, (h) adsorbent such as kaolin and bentonite, and (i) lubricant, such as talcum, calcium stearate, hard Fatty acid magnesium, solid polyethylene glycol, lauryl sodium sulfate and their mixture.Capsule, tablet and the pill the case where Under, dosage form also may include buffer.

The solid composite of similar type is also used as using such as lactose (lactose) or lactose (milk ) and the filler in the soft filling of excipient such as high molecular weight polyethylene glycol and the gelatine capsule filled firmly sugar.It can use Other coatings, prepare tablet, dragee, capsule, pill known in coating and shell, such as enteric coating and pharmaceutical-formulating art With the solid dosage forms of granule.

It should be understood that, it is contemplated that patient to be treated is selected the exact dose of PSMA- targeting particle by single doctor, is led to Often, adjustment dosage and application are to provide a effective amount of PSMA- targeting particle to treated patient.As used herein, PSMA target Refer to amount necessary to causing desired biological respinse to " effective quantity " of particle.As managed by those of ordinary skill in the art Solution can change PSMA- targeting according to factors such as all biological terminals as desired, drug to be delivered, target tissue, administration method The effective quantity of particle.For example, the effective quantity of the PSMA- targeting particle containing anticancer drug can be through desired amount through it is expected Period cause diameter of tumor reduce amount.Admissible other factor includes the seriousness of morbid state；It is treated Age, weight and the gender of patient；Diet, administration time and frequency of administration；Pharmaceutical composition；Reaction sensibility；And to treatment Tolerance and response.

Nano particle of the invention can be configured to the dosage unit form convenient for application and dose uniformity.It is used herein Statement " dosage unit form " refer to the physically separated unit of the nano particle suitable for patient to be treated.However, Ying Li Solution, the daily usage of composition of the invention should be determined in reliable medical judgment scope by attending physician.For appointing What nano particle, can initial estimation be controlled in cell culture assays or in the animal model of usually mouse, rabbit, dog or pig Treat effective dose.Animal model also is used to obtain desired concentration range and administration method.It is then possible to which this type of information is used In useful dosage and approach that measurement is applied in the mankind.It can be by standard pharmaceutical procedures in cell culture and/or experiment The therapeutic effect and toxicity that nano particle is determined in animal, for example, ED50 (treating effective dosage in 50% group)) and LD50 (the lethal dosage of 50% group).The dose ratio of toxicity and therapeutic effect is therapeutic index, and it can be expressed as The ratio of LD50/ED50.The pharmaceutical composition for showing big therapeutic index can be used in some embodiments.It is trained from cell The data obtained in nutrient analysis and zooscopy can be used for preparing a series of dosage used for the mankind.

The present invention also provides the kits comprising any above-mentioned composition, optionally have by as discussed previously Any suitable technology or via it is also known that drug delivery route apply the specification of any composition as described herein, The technology for example takes orally, is intravenous, pumping or implanted delivery apparatus." specification " can define the ingredient of popularization, and lead to Often it is related to the packaging or associated written explanation to composition of the invention.Specification can also include mentioning in any way Any oral or electronics explanation supplied." kit " usually definition includes any composition of the invention or group of the invention The combination of object and the packaging of specification are closed, it is also possible to include composition and any type of specification of the invention, institute State that specification is related to composition so that clinical professional will readily recognize that specification is and particular composition is related Mode provided.Kit described herein can also contain one or more containers, can contain the present composition And other compositions as discussed previously.Kit can also be containing for mixing in some cases, diluting and/or applying this hair The specification of bright composition.Kit can also comprising with one or more solvents, surface active ingredient, preservative and/ Or diluent (for example, physiological saline (0.9%NaCl) or 5% dextrose) other containers and for mixing, diluting or apply With the component in sample or the container applied to the object for needing this to treat.

The composition of kit can be provided as any suitable form, for example, as liquid solution or as drying Powder.When provided composition is dried powder, composition can be redissolved by adding suitable solvent, this can also be mentioned For.In the embodiment of composition for wherein using liquid form, liquid form can be concentration or instant.Solvent It will depend on the mode of nano particle and use or application.Suitable solvent for pharmaceutical composition is well-known (example It can get as discussed previously) and in the literature.Solvent will depend on the mode of nano particle and use or application.

In a further aspect, the invention further relates to the applications for promoting any nano particle as described herein.In some implementations In scheme, one or more compositions of the invention are used to prevent via the application popularization of any composition of the invention Or the various diseases for the treatment of, such as disease as described herein.As used herein, " popularization " includes related with composition of the invention All methods for carrying out commercial operation, including teaching method, hospital's introduction and other clinic introductions, the drug including drug product Industry activity and any advertisement or other popularization activities including any type of Written Communications, world-of-mouth communication and electronic communication.

Following embodiment is intended to explain certain embodiments of the present invention, but does not illustrate full scope of the invention.

Embodiment

The present invention is further illustrated by following embodiment.Embodiment should not be construed to further limit.

Embodiment 1: the formation of the liposome-CAP micella with neutral-surface.

Using two methods to generate micella: (1) film hydration effect and (2) quickly mix.It is acted on by film hydration, it will 215.6uL CAP (5mg/ml) and 117.4uL DPPC (bis- palmityl-sn- glyceryl -3- phosphocholine of 1,2-) (25mg/ ) and 329.9uL DPPE-PEG (1,2- distearyl acyl group-sn- glyceryl -3- phosphoethanolamine-N- [methoxyl group (poly- second two ml Alcohol) -2000] (25mg/ml) addition is together.Mixture is sufficiently mixed, evaporation is placed in vacuum at least 2 under argon gas stream Hour.Then, by mixture and water rehydration and ultrasonic treatment 10 minutes.Method by quickly mixing, by CAP, DPPC and Pegylated lipids DPPE-PEG is dissolved in ethyl alcohol and is mixed in MIVM with DI water rapidly according to molar ratio with different volumes. The following describe representative conditions.CAP (2mM), DPPC (2.67mM) and DPPE-PEG (2mM) are dissolved in ethyl alcohol CAP.By solution Loading is mixed into 5mL gastight syringe and rapidly with other three streams, and the stream is DI water.By CAP/ phospholipid solution One of stream and water flow are pumped with 6ml/min.Other two water flows are pumped with 54ml/min.Micella is suspended liquid cooling It is lyophilized dry 48 hours and is resuspended in aqueous solution.The molar ratio of CAP, DPPC and DPPE-PEG are shown in Table 2.

The information of the micella of table 21.CAP, DPPC and DPPE-PEG

The diameter (Fig. 7) of these micellas is measured using DLS.The average diameter of micella is reported in Table 2 below.Use dialysis process CAP is measured from the release in micella.Micella hinders the release of CAP to a couple of days (Fig. 8).

Also using other phosphatide to prepare lipid micelle.The other examples of the lipid micelle of test are with following molar ratio L- α-phosphatidyl choline (L- α-PC), DPPC-PEG and CAP combination.Diameter distribution is shown in Figure 9.

The information of the micella of table 3.CAP, DPPC and DPPE-PEG

Releasing result is shown when compared with the release of pure CAP, and release slows down (Figure 10).

Embodiment 2: the formation of the liposome-CAP micella with positively charged surface.

Using two methods to generate micella: (1) film hydration effect and (2) quickly mix.It is acted on by film hydration, it will DOTAP (1,2- dioleyl-3-trimethylammonium-propane) and CAP are dissolved in chloroform into 10mM concentration.By isometric DOTAP Mixed with CAP (500ul) solution and evaporate mixture under argon gas stream, be subsequently placed under vacuum at least 2 hours with Ensure solvent evaporating completely.Then, by desciccator diaphragm and 1ml DI water rehydration and ultrasonic treatment 10 minutes.Pass through what is quickly mixed Method is used for the cationic phospholipid of such as DOTAP to prepare micella with CAP.CAP (2mM) and DOTAP (2mM) are dissolved in ethyl alcohol In.Solution stowage is mixed in MIVM into 5mL gastight syringe and rapidly with other three streams, the stream is DI water.It will One of the stream of CAP/ phospholipid solution and water flow are pumped with 6ml/min.Other two water flows are pumped with 54ml/min. Micella suspension is freeze-dried 48 hours and is resuspended in aqueous solution.The diameter distribution of micella is shown in Figure 11.CAP from Release in DOTAP-CAP micella is shown in Figure 12.

The information of table 4.DOTAP-CAP micella.

Polymer-lipid body hybrid particles

Liposome-CAP micella is further wrapped up in the polymer by layer by layer deposition process to change particle surface Matter obtains preferable stability, controls particle diameter, and also maintain release.This method is fully able to control from several nanometers To hundreds of nanometers of particle diameter and be designed to negative, positive or neutral surface charge.The life that FDA for preparation ratifies The list of the compatible and biodegradable polymer of object is listed in Table 5 below.

The list of the biodegradable polymer of 5. anion of table and cation

Anionic biopolymer	Cationic biopolymers
		Alginates	Chitosan
Polyphosphate	Epsilon-polylysine
		Pectin (under weakly acidic solution)	Glucan
Carragheen	Poly- (carboxylic acid amide esters)
		Hyaluronic acid (HA)	Poly- (- L-lysine) (PLL)
Polyacrylic acid
		Poly- (lactic-co-glycolic acid (PLGA), acid blocked
Polylactic acid (PLA)
		Polyglycolic acid (PGA)
Poly-L-lactide -co- 6-caprolactone (PLCL)
		Poly- (acrylic acid-co-maleic acid)
Poly- (succinic acid-butanediol ester)
		Poly- (alkylcyanoacrylate) (PAC)

Embodiment 3:polyP-DOTAP-CAP nano particle

Prepare DOTAP-CAP micella as described in example 2 above first.By polyphosphate, (polyP averagely has 75 weights Multiple unit) 5mM aqueous solution is made.The DOTAP-CAP micella of resuspension is mixed with polyP solution under various ratios.In polyP The typical DLS measurement of particle diameter before and after deposition is shown in Figure 14.

The information of table 6.polyP-DOTAP-CAP nano particle.

The stability for monitoring polyP-DOTAP-CAP nano particle at 4 DEG C and at room temperature continues five days (Figure 15).? The zeta potential of all three samples slightly reduces during 5 days.The diameter of particle remains unchanged in five days at 4 DEG C, in room temperature The diameter of lower particle increases according to power-law function.

CAP has significant duration (Figure 16) from the release in these polyP-DOTAP-CAP nano particles.

Embodiment 4:PEG-PAA-DOTAP-CAP nano particle.

In order to enhance spatial masking effect and extend the circulation of nano particle in blood, we devise di-block copolymer Object PEG-b-PAA.Preparation DOTAP-CAP micella as described in example 2 above.PEG-PAA is soluble in water at 0.6mM.By 500uL DOTAP-CAP micella is mixed with isometric PEG-PAA solution.The structure of PEG-PAA-DOTAP-CAP nano particle is shown in figure In 17.

The information of table 7.PEG-PAA-DOTAP-CAP nano particle suspension.

Zeta potential is monitored at room temperature and diameter distribution continues five days.As shown in Figure 19, the zeta potential of PEG-PAA is in Property and particle keep stablize.

After seven days, CAP is shown in Figure 20 from the sustained release in PEG-PAA-DOTAP-CAP nano particle.

Equivalent form

Those skilled in the art will appreciate that or can determine the specific of invention as described herein using only routine experiment Many equivalent forms of embodiment.It is intended to cover such equivalent form in claim.

It is incorporated by reference into

All patents recited herein, disclosed patent application, website and other bibliography full content lead to accordingly Reference is crossed to be expressly incorporated herein with its entirety.

Bibliography

1.K.Chudasama“A review on capacetabine”World Journal of Pharmaceutical Research(2015),4(7),1427-1432.

2.S.Lam,H.Guchelaar,E.Boven“The role of pharmacogenetics in capecitabine efficacy and toxicit”Cancer Treatment Reviews(2016),50,9-22.

3.J.Kwakman,C.Punt“Oral drugs in the treatment of metastatic colorectal cancer”Expert Opinion on Pharmacotherapy(2016),17(10),1351-1361.

4.H.Iqbal,Q.Pan“Capecitabine for treating head and neck cancer”Expert Opinion on Investigational Drugs(2016),25(7),851-859.

5.R.Mahlberg,S.Lorenzen,P.Thuss-Patience,V.Heinemann,P.Pfeiffer, M.Moehler“New Perspectives in the Treatment of Advanced Gastric Cancer:Oral 5-FU Therapy in Combination with Cisplatin”Chemotherapy(Basel,Switzerland) 2016,62(1),62-70.

Claims

1. being used for the composition comprising nano particle for the treatment of cancer, the nano particle includes:

Phosphatide core, it includes one or more phosphatide and at least one of capecitabines and its active metabolite；And

At least one layer of one or more polymer on phosphatide core surface,

Wherein when providing to object, the composition provides capecitabine or the sustained release of its active metabolite.

2. composition as described in claim 1, one or more of them lipid includes at least one below: 1,2- bis- caprinoyl Base-sn- glyceryl -3- phosphocholine (DDPC), 1,2- dilauroyl-sn- glyceryl -3- phosphoethanolamine (DLPE), two Myristoyl phosphatidyl choline (DMPC), dimyristoylphosphatidylglycerol (DMPG), bis- myristoyl-sn- of 1,2- Glyceryl -3- phosphoethanolamine (DMPE-PEG), 1- palmityl -2- myristoyl-sn- glyceryl -3- phosphocholine (PMPC), bis- palmityl-sn- glyceryl -3- phosphoric acid-(1 '-rac- glycerol) (DPPG) of 1,2-, bis- palmityl-sn- of 1,2- Glyceryl -3- phosphocholine (DPPC), bis- palmityl-sn- glyceryl -3- phosphoethanolamine (DPPE-PEG) of 1,2-, 1,2- Two palmityl-sn- glyceryl -3- phosphate (sodium salt) (DPPA), 1- palmityl -2- oleoyl-sn- glyceryl -3- phosphorus Sour choline (POPC), 1- palmityl -2- stearyl-sn- glyceryl -3- phosphocholine (PSOC), 1- stearyl -2- palm fibre Palmitic acid docosahexaenoyl-sn-glycero -3- phosphocholine (SPPC), 1,2- distearyl acyl group-sn- glyceryl -3- phosphoethanolamine-N- [first Oxygroup (polyethylene glycol) -2000] (DEPE-PEG), 1,2- distearyl acyl group-sn- glyceryl -3- phosphoethanolamine-N- [hexichol And cyclooctyl (polyethylene glycol) -2000] (DSPE-PEG), L- α-phosphatidyl choline (L- α-PC), bis- sub-oleoyl-sn- of 1,2- Glyceryl -3- phosphocholine (DLPC), 1,2- dioleoyl-sn- glyceryl -3- phosphoric acid-(1 '-rac- glycerol) (DOPG), 1, 2- distearyl acyl group-sn- glyceryl -3- phosphoric acid-(1 '-rac- glycerol) (DSPG), 1,2- distearyl acyl group -3- trimethyl ammonium - Propane (DSTAP), 1,2- dioleyl-3-trimethylammonium-propane (DOTAP), 1,2- dioleoyl-sn- glyceryl -3- phosphoric acid Salt (DOPA), 1,2- dioleoyl-sn- glyceryl -3- phosphoethanolamine (DOPE), 1- stearyl -2- oleoyl-sn- are sweet Oil base -3- phosphocholine (SOPC), 1,2- dioleoyl-sn- glyceryl -3- phosphocholine (DOPC).

3. composition as described in claim 1, wherein one or more polymer include at least one below: poly- (lactic-co-glycolic acid) (PLGA) or its PEGylated forms PEG-PLGA, polylactic acid (PLA) or its Pegylation shape Formula PEG-PLA, polyglycolic acid (PGA) or its PEGylated forms PEG-PGA, poly-L-lactide -co- 6-caprolactone (PLCL) or its PEGylated forms PEG-PLCL, hyaluronic acid (HA), polyacrylic acid (PAA) or PEG-PAA, polyphosphate (polyP), poly- (acrylic acid-co-maleic acid), poly- (succinic acid-butanediol ester), poly- (alkylcyanoacrylate) (PAC) or its PEGylated forms PEG-PAC.

4. composition as described in claim 1, wherein the composition also includes selected from anticancer drug, antibiotic, antiviral In agent, antifungal agent, pest repellant, nutrient, small molecule, siRNA, antioxidant and antibody or traditional radioactive isotope extremely A kind of few active constituent.

5. composition as described in claim 1, one or more of them lipid includes: 1,2-, bis- palmityl-sn- glycerol Base -3- phosphocholine (DPPC), bis- palmityl-sn- glyceryl -3- phosphoethanolamine (DPPE-PEG) of 1,2-, L- α-phosphatide Phatidylcholine (L- α-PC), bis- sub-oleoyl-sn- glyceryl -3- phosphocholine (DLPC) of 1,2- or 1,2- dioleoyl -3- front three Base ammonium-propane (DOTAP),

Wherein the common molar ratio of the lipid and pegylated lipids is 100:0 to 50:50, and saturation lipid and insatiable hunger Molar ratio with lipid is usually 100:0 to 25:75.

6. composition as described in claim 1 also includes at least one targeting agent, wherein the targeting agent makes the nanometer The tissue/cell of illness is targeted to Selective feeding, so that whole-body dose be made to minimize.

7. composition as described in claim 1 also includes at least one targeting agent, wherein the targeting agent include antibody or Its function fragment, small molecule, peptide, carbohydrate, siRNA, protein, nucleic acid, aptamer, the second nano particle, cell because Son, chemotactic factor (CF), lymphokine, receptor, lipid, agglutinin, ferrous metal, magnetic-particle, connexon, isotope and they Combination.

8. composition as described in claim 1, wherein the nano particle has the diameter of 10nm to 200nm.

9. composition as described in claim 1, wherein the bioavilability of composition increases, selectively such as nausea, vomit It spits, dermatitis, bone marrow suppression, one or more side effects reduction of cardiac toxic and diarrhea, and capecitabine or its active generation It thanks to object to be discharged in a continuous manner.

10. composition as described in claim 1, wherein the composition is suitable for intramuscular, subcutaneous, intravascular or intravenously applies With.

11. the method for forming nano particle comprising:

The combination of one or more phosphatide, one or more solvents and at least one of capecitabine and its metabolin is come Form organic phase；

It combines one or more targeting agents to form lipid water phase with water；

It mixes the organic phase in multiple entry vortex mixer with the water phase and generates suspension, oneself of micella thus occurs Assembling；

It is spray-dried or is freeze-dried suspension and recycle the organic solvent；And

It mixes the solution with one or more polymer with the micella, layer-by-layer polymer deposits thus occurs, thus shape At nano particle,

And wherein when providing to object, the nano particle is capable of providing the sustained release of capecitabine and its metabolin.

12. the method for forming composition described in claim 1, wherein preferably diameter preparation has with particle diameter repeatability Control the nano particle of good physicochemical properties such as surface nature.

13. the method for treating the doubtful patient with disease comprising application nano particle, wherein the nano particle packet Contain: the phosphatide core comprising at least one of one or more phosphatide and capecitabine and its active metabolite；With in phosphatide At least one layer of one or more polymer on core surface,

Wherein when providing the nano particle to object, at least one of the capecitabine and its active metabolite cause One or more side effects reduce, the side effect such as Nausea and vomiting, dermatitis, bone marrow suppression, cardiac toxic and diarrhea.

14. method as claimed in claim 13, wherein apply the nano particle include by intramuscular, subcutaneous, intravascular or It intravenously applies and applies the nano particle.

15. method as claimed in claim 13, wherein the disease is selected from tumor disease, the nervous system disease and metabolism Disease.

16. method as claimed in claim 13, wherein the disease is selected from Parkinson's disease, Alzheimer's disease, multiple Property hardening, ALS, sequelae, behavior and cognitive disorder, self-closing disease spectrum, depression and tumor disease.

17. method as claimed in claim 13, wherein at least one of the capecitabine and its active metabolite are to hold Continuous mode is discharged.

18. pharmaceutical agent, it includes:

For the nano particle of drug delivery, it includes cause such as Nausea and vomiting, dermatitis, bone marrow suppression, cardiac toxic and abdomen At least one of capecitabine and its active metabolite of one of the side effect rushed down and encapsulation phosphatide and the capecitabine And its at least one layer of one or more polymer of at least one of active metabolite, and the reagent does not cause one kind Or a variety of side effects.

19. the method for treating the doubtful patient with disease, the method includes applying nano particle, wherein the nano particle Include:

Phosphatide core comprising at least one of one or more phosphatide and capecitabine and its active metabolite；With

At least one layer of one or more polymer on phosphatide core surface.

20. the method for treating the doubtful object with cancer comprising:

Identify the doubtful patient with cancer；And

At least one in the capecitabine and its metabolin for the amount for being enough to mitigate the cancer in the object is provided to the object Kind, wherein at least one of the capecitabine and its metabolin are present in nano particle, the nano particle includes:

Phosphatide core comprising at least one of one or more phosphatide and the capecitabine and its active metabolite；With At least one layer of one or more polymer on phosphatide core surface,

Wherein when providing to object, the nano particle does not cause to include Nausea and vomiting, dermatitis, bone marrow suppression, cardiac toxic With one of the side effect including diarrhea.

21. method as claimed in claim 20, wherein the cancer is breast cancer, colorectal cancer or cancer of pancreas.