CN101123949B - Medicine lipid composition - Google Patents

Abstract

The present invention relates to a particulate composition containing; a) 5 to 90% of at least one phosphatidyl choline component b) 5 to 90% of at least one diacyl glycerol component, at least one tocopherol, or mixtures thereof, and c) 1 to 40% of at least one non-ionic stabilising amphiphile, where all parts are by weight relative to the sum of the weights of a+b+c and where the composition contains particles of at least one non-lamellar phase structure or forms particles of at least one non-lamellar phase structure when contacted with an aqueous fluid. The invention additionally relates to pharmaceutical formulations containing such compositions, methods for their formation and methods of treatment comprising their administration.

Description

Medicine lipid composition

Technical field

The present invention relates to the protection, dissolving, stable and send of activating agent in pharmaceutical composition and type medicament nutriment (neutraceutical) compositions.Especially, the present invention relates to amphiphilic compositions and preparation, and send system based on these activating agent.

Background technology

In the sending of many materials, especially in the body of human body or animal body, sending, demonstrate sizable potential based on the preparation of amphiphilic substance.Assemble polar group and the non-polar group that forms polarity and nonpolar district because amphiphilic substance has, it can dissolve polarity and non-polar compound effectively.In addition; The many structures that in polarity and/or non-polar solven, form through amphiphilic substance/structural agent (structuring agent) have sizable polar/non-polar frontier district, can absorb in said polar/non-polar frontier district and other amphipathic compound of stabilisation.

In amphiphilic substance/water, amphiphilic substance/oil and amphiphilic substance/oil/water figure, the formation of non-layer-like zone is known phenomenon.Like this comprise liquid crystalline phase and L mutually ₃" sponge " phase, said liquid crystalline phase for example be a cube P, cube D, cube G and hexagonal mutually, it is fluid on molecular level, but shows tangible long-range order, said L ₃" sponge " comprises the three-dimensional co-continuous network of many places interconnection double-layer tablet (bilayersheets) mutually, and it lacks the long-range order of liquid crystalline phase.Depend on their bending, these can be described to positive (clinodactyly is towards nonpolar district) or anti-phase (clinodactyly is towards doping region) mutually.The spontaneous bending of lipid system near zero situation under, structure typically is stratiform, single or multiple lift vesicle/liposome for example, spontaneous bending be more minus or the situation of corrigendum under, micelle, cube phase and hexagonal typically prevail mutually.

Non-stratiform (for example liquid crystal and L ₃) be thermodynamic stable system mutually.That is to say that they are not simply will separate and/or cambium layer, lamellar phase or similarly metastable state again, but the Thermodynamically stable form of mixture.

After deliberation layered system and non-layered system as food, cosmetics, nutriment, diagnostic agent and the carrier of medicament and/or the characteristic of excipient; And with regard to the doping region and the high internal surface area between the nonpolar district of non-layered system, think that they have sizable advantage.This has caused the considerable research of how non-lamellar phase, especially at controlled release preparation be used to dissolve aspect the chemical compound of relative low solubility.

As discussed above, the non-stratiform bulk phase of body (bulk) is typical thermodynamic stable system.In addition, this this bulk phase can be dispersed in polarity or the non-polar solven in body solvent (bulksolvent), to form the particle of non-stratiform (especially liquid crystal) phase.This have use the body immiscible phase with the situation that has problems in for example in parenteral application, the advantage of the non-lamellar phase of applied ontology.Also can realize the further control of chemical compound release characteristic through the dispersion of so non-booklike particle.

Liquid crystal or L ₃Mutually can be in thermodynamical equilibrium or near thermodynamical equilibrium, and can be scattered in the colloidal stabilising dispersions of non-booklike particle with excessive solvent.Such particle can be that (being thermodynamics) is stable fully, or can progressively degrade, thereby the release characteristic of the activating agent that control is therewith prepared.The formation of dispersion can be spontaneous or for example shear or ultransonic result as mechanical force.These non-booklike particles have sizable meaning in activating agent is sent, and have been suggested the carrier as many such active matters.

The method that for example forms the dispersed particle of non-lamellar phase at solvent in the water is described in US5, in 531,925.Such particle has non-layered liquid crystal or L ₃Inner phase and stratiform or L ₃The surface mutually and can comprise active component.

Through for example to liquid crystal or L ₃Add the surface in the inner phase and form agent solution mutually, stir, can form known liquid crystal or L to form coarse dispersion and to smash the method for gained mixture ₃The particle of inner phase.

In order to evaluate the existence of liquid crystalline phase, the liquid crystal material of small angle X-ray diffraction capable of using (SAX), freezing transmission electron microscope (cryo-TEM) or nuclear magnetic resonance, NMR (NMR) spectrum research test expection.Can especially utilize the granularity and the particle size distribution of laser light scattering instrument test dispersed particle through light scattering.

Dispersion, the especially intravenous administration of hoping to contain active component give human body or animal body those be colloidal, their granularity should not surpass 10 μ m in other words, especially is no more than 5 μ m, particularly is no more than 1 μ m.If disperse intravital particle to surpass this size, this dispersion possibly not be that colloidal state is stable so, when the said preparation of intravenous administration, has sizable risk that causes thromboembolism.And, hope narrower particle size distribution, farthest to control the release of any activating agent.In desire through except intravenous method (for example oral, intramuscular, subcutaneous, rectum or through sucking) when using particle composition; Said particle is unnecessary to be colloidal; But provide well-characterized and reproducible particle size distribution remain favourable so that control the speed that said particle decomposes and/or activating agent discharges.

The granularity of particle composition also should be stable, to store considerable time.If the particle size distribution significant change may influence the effective transfer rate diffusion and the rate of release of any activating agent (for example, because) of compositions so unfriendly.The colloidal dispersants particle size stable property that is used for intravenous administration remains the problem of paying close attention to.If the particle size distribution of such dispersion unstable (for example for storage and distribution) is passed so in time and possibly formed big particle, and is dangerous when using.Even be not that direct danger is arranged, the storage unstability also can cause the significant change of pharmacokinetics, kinetics and/or effectiveness.

Except controlling particle size, hope that maximization is in the ratio of the particle in the non-lamellar phase that needs, with seal with regard to load capacity, protectiveness, speeches such as controlled release, reproducibility maximize its useful effect.Therefore should make booklike particle for example the ratio of single or multiple lift vesicle minimize.

The method of the dispersed particle of the non-lamellar phase of known formation is efficiently, but produces " impurity " stratiform vesicle particle of wide relatively particle size distribution and suitable vast scale usually.The energy input that the ratio of broken (fragmenting) agent and/or stabilizing agent (for example surfactant, copolymer and/or protein) or increase homogenize are handled in the increase preparation can be used for making size distribution narrow but cost is the ratio increase of booklike particle.

A limitation of the non-laminar composition of at present available or suggestion is that they depend on lipid usually, and when high concentration, said lipid can not tolerate in vivo well.Especially, if when high concentration is used (particularly parenteral), (comprise that common glyceryl monooleate-GMO) can be deleterious, it is dose limitation property for the conventional monoacylglycerol that uses.The probability of the toxic and side effects of lipid carrier also can be limited to highly serious character (highly serous nature) with the indication scope of using activating agent, wherein can tolerate the risk of side effect.Therefore; Compare with the compositions of extensive use (for example comprise GMO those), shapable and as particle dispersion stable, show that the lipid composition that predictable non-lamellar phase behavior and toxicity reduce (for example visible from hemolytic index and/or studies on acute toxicity) will be very large progress.If such preparation is shapable and stable as the particle (the for example diameter of 0.05～about 2 μ m) of colloidal state size, and having narrow, monomodal grit distribution, will be further favourable so.Find to be difficult to the particular dispersing body that provides stable in the document, and have only lamellar dispersions stable as can to store (Kamo et al.Langmuir 19,9191-9195 (2003)) more than several days.

Summary of the invention

The inventor has confirmed to contain the mixture of at least 3 kinds of amphiphilic components of DG (DAG), tocopherol or diacyl PHOSPHATIDYL ETHANOLAMINE (PE) component or its mixture, phosphatidylcholine (PC) component and nonionic stabilising component consisting unexpectedly; Stable non-lamellar dispersions can be efficiently formed, and surprising hypotoxicity can be demonstrated in vivo.

First aspect, therefore the present invention provides particle composition, and it comprises:

A) 5 to 90% at least a phosphatidylcholine component,

B) 5 to 90% at least a DG component, at least a tocopherol or its mixture and

C) at least a nonionic stability amphiphilic substance of 1 to 40% (preferred 2 to 40%),

Wherein all umbers are with respect to the gross weight of a+b+c by weight, and wherein compositions comprises the particle of at least a non-lamellar phase structure or when contacting with aqueous fluid, forms the particle of at least a non-lamellar phase structure.

Preferred composition of the present invention comprises at least a as activating agent described herein in addition, and can comprise solvent (especially water or aqueous solvent or solvent mixture).Said compositions also can comprise appropriate carriers, excipient, filler, stabilizing agent and similar component.

Aspect further, the present invention provides pharmaceutical preparation, and said pharmaceutical preparation comprises carrier or the excipient that at least a compositions of the present invention and at least a medicine can tolerate.

Aspect further, the present invention provides the treatment human or animal method of object, comprises and uses the optional compositions of the present invention that comprises activating agent.In this regard, said Therapeutic Method is especially treated the method for inflammation and/or stimulation (irritation), especially in body cavity, for example in gastrointestinal tract.

Another further aspect, the present invention provides the purposes of compositions of the present invention in treatment, the especially optional present composition that comprises activating agent is used for treating the purposes in the medicine of inflammation and/or stimulation (especially body cavity for example gastrointestinal tract) in preparation.

Ternary amphiphilic compositions of the present invention comprise at least a PC component (component a), at least a DAG, at least a tocopherol and/or at least a PE component (components b) and at least a nonionic stability amphiphilic component (amount of component b).Amount of component b will promote the fragmentation of compositions especially.

At least 5% of amphiphilic component (a+b+c) gross weight should be component a.Preferred ingredient a will be 5 to 50%, and more preferably 10 to 40%.Correspondingly, components b will be at least 5% of an a+b+c weight, and will be preferred 20 to 85%, more preferably 30 to 75%.Amount of component b should be 2 to 40% of an a+b+c gross weight, and is preferred 3 to 35%, more preferably 5 to 30%.

In ternary amphiphilic compositions, phosphatidylcholine component " a " is mainly formed with the lipid of two nonpolar acyl chains that are connected through ester bond by having the phosphatidylcholine polar group.As in the document, this component refers to phosphatidylcholine (PC) in this article and can be made up of a kind of pure compound, for example synthetic dioleyl phosphatidyl choline or, be more preferably the mixture of PCs that PCs for example comes from the natural source of purification.PC is particularly advantageous component, this be because it be extensively and obtain easily and purification from multiple natural source easily.The present invention can comprise blended PCs useful effect with the product of natural source; Be superior to some other lipid composition significantly; Dioleoyl PHOSPHATIDYL ETHANOLAMINE (DOPE) for example; The extraction of said dioleoyl PHOSPHATIDYL ETHANOLAMINE and purification more complicacy maybe must be synthetic, and this makes them more be difficult to industrial-scale production and obtain.

Typically, the PCs that extracts from natural source will have the mixture of acyl chain, and this mixture will have some variations according to the tissue that obtains extract.Liver PC for example has the acyl chain length (C at least of relative wide region ₁₆To C ₂₀), and have the saturated acyl group of larger proportion and acyl group with 2 above degrees of unsaturation.By contrast, Semen sojae atricolor PC typically mainly comprises the C that contains 0 or 2 degree of unsaturation ₁₆To C ₁₈Acyl group.This makes the characteristic of the present composition to change subtly through selecting suitable substance P C component or its mixture.Preferred PCs comprises the PC (SPC) of ovum, heart, brain, liver, especially Semen sojae atricolor.When needing the saturated PCs of higher proportion, also hydrogenatable PC or its any part.

Because PC component of the present invention is natural extract preferably, so there is a spot of non-PC " impurity " usually.For the lipid content with other polar group, the accurate level of purification of PC component will depend on the concrete application of the present composition to be used.Important factor is keep the phase behavior of said compositions, very high stability and low-down toxicity through the component of selecting adequate purity.As long as these performances remain acceptable, the purity of PC will have less relatively importance so.Yet as general guidance, the PC component contains the non-PC polar group that is no more than 10wt% usually.In some embodiments, non-PC polar group preferably is no more than 5wt%, more preferably no more than 2wt%.

In compositions of the present invention, components b) be DG (DAG) and/or tocopherol.These can be by single, pure DG, or tocopherol is formed; It can be the mixture of DG and/or tocopherol; It maybe can be natural extract with purification of high-load DG and/or tocopherol.The carboxyl groups of preferred DG has 10 to 24 carbon atoms, preferred 12 to 20 carbon atoms, most preferably 14 to 18 carbon atoms independently.Saturated and/or undersaturated carboxyl groups is suitable, but preferably has the group of one, two or three two keys.Said carboxyl groups can be identical or different.Highly preferred DAG is NOFABLE GO-902P (GDO) and composition thereof.

Term used herein " tocopherol " is used to represent nonionic lipid tocopherol, is commonly referred to vitamin E, and/or its any suitable salt and/or analog.Suitable analog will be have phase behavior, stability and lack toxic those, these performances are characteristics of the present composition, and in water, do not form the liquid crystalline phase structure usually as pure compound.Most preferred tocopherol is the tocopherol itself with following structure.Obviously, especially from natural source during purification, possibly have the non-tocopherol of fraction " impurity " but this will be not enough to change favourable phase behavior, stability and avirulence.Typically, tocopherol will comprise the non-tocopherol anologs chemical compound that is no more than 10wt%, preferably be no more than 5wt%, and be most preferably not exceeding 2wt%.

Tocopherol

For the PC component, components b, for example DG, and/or tocopherol, can be used as natural extract provides.For the availability and reliability of material, this has significant advantage.Yet the DAG component is a natural extract, possibly still exist a spot of non--the DAG lipid.For the PC component, will be that compositions has favourable stability of the present invention and avirulence for the vital test of DAG component purity.Typically the DAG component will have other lipid that is no more than 15wt%, preferably be no more than 10% and more preferably no more than 5%.DAGs (as described herein) is preferred ingredients b).

The preferred compositions of composition components b) is the mixture of at least a DAG (for example GDO) and at least a tocopherol.Such mixture comprises 2: 98 to 98: 2 by weight tocopherol: GDO, for example 10: 90 to 90: 10 tocopherol: GDO, especially these chemical compounds of 20: 80 to 80: 20.

The preferred especially combination of component a and b is PC and DAG, and wherein two kinds of components have 50%C18 at least: 1 (oleoyl) and/or 50%C18: 2 (inferior oleoyl) acyl group.Semen sojae atricolor PC and ovum PC are special preferred examples.These preferred weight ratio is at 1: 5 to 3: 2, and most preferably 2: 5 to 4: 5PC: GDO.Bioactive a kind of measurement of lipid is its dissolubility in water or aqueous solution.Component with high relatively water solubility is kept the monomeric higher equilibrium concentration of dissolved lipid in solution, this can be the reason of observed biological effect at least in part." the single oleic acid glycerol " used always (GMO) for example, at room temperature has 10 ^-7The equilibrium water dissolubility of the M order of magnitude, and bigger under physiological temp.Relatively, preferred DG and diacyl PHOSPHATIDYL ETHANOLAMINE at room temperature can have and be no more than 10 ^-8Or more typically 10 ^-9The dissolubility of M, preferred 5 * 10 ^-10M and more preferably 10 ^-10M or still less.Minimum desirable dissolubility common about 10 ^-15M.Especially, when high dilution, the stability of non-layered system will depend on that lipid molecular leaves the structural material surface and is diffused into the speed in the solution.Therefore, the stability of non-booklike particle dispersion will be directly with solvent in monomeric dissolubility relevant.

Amount of component b plays cracking agent, and helps control and the stability and the promotion of particle phase behavior and stablize non-lamellar phase to be broken into particle.Amount of component b will exist with the level that enough causes the broken and/or stable broken non-lamellar phase particle of compositions.Such fragmentation can be that shearing and/or ultrasonic is for example passed through in the spontaneous physics fragmentation that maybe possibly need.Consider the embodiment of this paper, those skilled in the art will have no to estimate any compositions difficultly and whether comprise enough cracking agents.

Generally speaking, nonionic stability amphiphilic substance " c " is the component that improves dispersion stability, especially as the dispersion of colloidal particle.The preferred form of these nonionic amphiphilic substances is the nonionic lipids that are grafted with PEO and/or PPOX chain (or its copolymer).

Such chemical compound is the for example grafted fatty acid of polyalkylene oxide or substituted fatty acid (especially hydroxylating fatty acid), the grafted lipid of polyalkylene oxide or polyol, and said polyol has and is grafted to one or more (all preferred) polyalkylene oxide groups of alcohol moiety and one or more fatty acid chains with the opposite ends that is connected to one or more said polyalkylene oxide chains.Instance comprises stearic acid Polyethylene Glycol (PEG) ester; Distearyl acid PEG ester; Lauric acid PEG ester; Oleic acid PEG ester; The Oleum Ricini of polyethoxylated; PEG-DOPE; PEG-(4-hydroxy stearic acid ester) (Solutol); PEG-sorbitan-monolaurate (Polysorbate 20 or 21); PEG-sorbitan-monopalmitate (Polysorbate 40); PEG-sorbitan-monostearate (Polysorbate 60 or 61); PEG-sorbitan-tristearate (Polysorbate 65); PEG-sorbitan-monoleate (Polysorbate 80 or 81); PEG-sorbitan-trioleate (Polysorbate 85); PEG-sorbitan-single vaccenic acid ester (Polysorbate 120) and succinic acid d-alpha tocopherol base cetomacrogol 1000 ester (vitamin E TPGS).The PEG chain can suitably be connected on other amphiphilic substance component through ester bond or ehter bond.Typically, the polyalkylene oxide total content of the molecule of amount of component b will be no more than 50 monomers, preferably be no more than 30 monomers.The Oleum Ricini of Polysorbates 20 and 80, solutol, succinic acid d-alpha tocopherol base cetomacrogol 1000 ester (vitamin E TPGS) and polyethoxylated most preferably.

The inventor has confirmed that now the stability of non-booklike particle dispersion demonstrates the sizable dependency to used stabilizer types.Especially, above-mentioned dispersant is preferred especially, this be because have been found that high-molecular weight surfactant to the stability of particle dispersion significantly less than stability that lower molecular weight compound showed.Therefore, in one embodiment, amount of component b) should comprise or preferably basically by or be lower than 10,000 dalton by molecular weight, preferably be lower than 8,000 dalton and form more preferably less than 5000 daltonian surfactants.Similarly, amount of component b) should preferably not comprise block copolymer surfactant, especially molecular weight and surpass the sort of of above-mentioned scope.This is surprising especially, is effective because the surfactant of higher molecular weight has demonstrated for stablizing the compositions related of stratiform form.

An importance of the present invention is that compositions can be mixed with the described preconcentrate that comprises cosolvent of this paper.These concentrate are particularly suited in parenteral application, being used as control delivery.In this purposes, the most available ratio between the c/a+b+c is 1 to 30% amount of component b, more preferably 3 to 25%.For the bank that the compositions that is intended to discharge at about 1 time-of-week forms, most preferred scope is 3 to 10%c.

The amphiphilic component of compositions of the present invention can be basically by, or only by component a), b) and c) form (, adding any activating agent) if amphipathic.In this embodiment, at least 95%, preferred at least 98% and most preferably in fact 100% amphiphilic component will be a kind of in these.Alternatively, in addition, optional, amphiphilic component d) can be in weight a)+b)+c)+d), have preferably at the most 8%, more preferably at the most 5% with 10% amount at the most.This component d) can be any suitable amphiphilic substance, for example natural or synthetic lipid, or derivatives thereof or analog.Special preferred ingredient d) be the ion-type lipid, fatty acid or its biology salt that can tolerate for example.

Compositions of the present invention comprises non-booklike particle or when contacting with aqueous fluid, forms the compositions of such particle.Such fluid can be that the fluid (for example water or Sterile Saline) for delivery to object maybe can be fluid, blood or the iuntercellular fluid of body fluid, especially stomach fluid, intestinal fluid, mucomembranous surface.

Term used herein " non-stratiform " is used for expression cube, hexagonal, L ₂Or L ₃Phase structure or its any combination, with as the layer structure in lamellar phase or liposome/vesicle, found opposite.When particle was described to have non-lamellar phase or structure, this represented that particle inside has this structure at least.Many such particles will have two different zones, interior zone and circumferential surface zone.Said surf zone, even in " non-stratiform " particle, can be lamellated or crystal, and it is mutually any to be the crystal layer that comprises high-sequential, liquid crystalline phase with in fact unordered fluid layer.

Term used herein " booklike particle " is meant vesicle shape particle (for example liposome), it is characterized in that they comprise the stratiform bilayer of one or more outsides of amphiphilic substance, around inner solvent compartment.

In one aspect of the invention, said compositions comprises non-booklike particle.This representes in the particle of the present invention (preferred colloidal state), at least 50%, preferably at least 75%, and most preferably at least 85% (as through cubing) is unstratified (for example combining freezing-TEM or the SAXS definite through laser diffraction).In an alternative aspect of the present invention, said compositions forms non-booklike particle when contacting with aqueous fluid.When this is illustrated in and contacts with aqueous fluid (as described herein), at least 50%, preferably at least 75%, most preferably at least 85% particle (as through cubing) becomes non-booklike particle.

In a preferred embodiment of the invention, compositions of the present invention comprises or forms hexagonal and/or L ₃The anti-phase particle.The most preferred group compound comprises or forms L ₃The phase particle.L ₃, be called the long-range order that " sponge " lacks real liquid crystalline phase mutually again, but form by the sheet of the double-layer of lipoid of many places interconnection, wherein these " interconnection " do not adopt being seen ordered arrangement in the cubic liquid crystal structure.

In an alternate particularly advantageous again embodiment, compositions of the present invention can form I ₂Or L ₂Non-lamellar phase.Said I ₂Be cubic liquid crystal anti-phase mutually with discontinuous water zone.This is advantageous particularly in the controlled release of activating agent, when especially for example the water dissolvable active matter combines with the polarity activating agent.Said L ₂Have similar advantage mutually and be in so-called " reverse micelle " mutually in, said " reverse micelle " has around the continuous hydrophobic district of discrete polarity nuclear mutually.

For many combinations of lipid, only there is some non-lamellar phase, or exists with any steady statue.A surprising characteristic of the present invention is that compositions described herein often shows non-lamellar phase, and said non-lamellar phase does not combine existence with many other of component.Therefore, in a particularly advantageous embodiment, the present invention relates to have the compositions of combination of components, there is I in the combination of said component when adopting the aqueous solvent dilution ₂And/or L ₂Phase region.Through with aqueous solvent diluted composition and the phase structure that gets through method institute described herein simply, can easily test and have or do not exist such zone in any particular combination.

When in compositions of the present invention, preparing activating agent, said activating agent has influence with common phase behavior to structural agent.For example, some activating agent (for example Ciclosporin A) is compared the bigger hogging bending of introducing with some structural agents, when high concentration, can cause the formation of height hogging bending phase, for example reverse micelle L ₂Mutually rather than cube or the hexagonal liquid crystalline phase.But, through preparing, can such activating agent be mixed with for example hexagonal anti-phase with the mixture of the component a with the spontaneous bending of less negativity, b and c.Through the method, whole mixt provides suitable hogging bending in compositions of the present invention, using.

Those skilled in the art can adopt the method for standard to estimate the spontaneous flexibility of any ad hoc structure agent mixture of component (or itself and other) or comprise the influence of activating agent to this.This can be through carrying out as follows, the body phase behavior through every kind of structural agent in the research water for example, and the concentration that changes contained activating agent is subsequently studied.Select the suitable mixture of component through any method shown in this paper (for example polarized light, SAXS, freezing-TEM etc.) with for every kind of situation, can analyze said phase.In some cases, at activating agent the phase behavior of mixture is influenced when remarkable, selected structural agent itself possibly not provide ideal non-lamellar phase (for example can have too little or too big spontaneous bending), but only when preparing with activating agent, can produce this mutually.Therefore, when adding activating agent, equilibrium phase can be from for example cube being varied to the hexagonal liquid crystalline phase.

One preferred aspect, compositions of the present invention comprises at least a activating agent.Suitable activating agent comprises human and veterinary drug and vaccine, diagnostic agent, and " alternate " activating agent is plants essential oil, extract or aromatic, cosmetic agent, nutrient, food additive etc. for example.Suitably the instance of medicine comprises for example for example polyene macrolides (for example amphotericin B) or triazole antifungal agent of beta-lactam or Macrocyclic peptides antibiotic, antifungal of antibacterial; Anticancer and/or antiviral drugs is nucleoside analog, paclitaxel and derivant for example, and anti-inflammatory agent is nonsteroidal antiinflammatory drug for example, and cardiovascular drugs comprises cholesterol reducing and hypotensor; Analgesic; Anesthetis, antidepressant comprise the serotonin absorption inhibitor, vaccine and bone regulator.Diagnostic agent comprises that radionuclide labelled compound and contrast agent comprise X-ray, ultrasonic and MRI contrast-enhancing agent.Nutrient comprises vitamin, coenzyme, food additive etc.Being used for activating agent of the present invention is not any component a described herein, b or c usually.Other preferred activating agent comprises insulin and insulin analog; Growth hormone is human growth hormone (hgh) for example; Immunosuppressant is tacrolimus and Ciclosporin A for example, peptide medicament for example described herein those, comprise Sandostatin LAR Depot, salmon calcitonin see calcimar, Desmopressin, somatostatin; Antibody and antibody fragment, nucleic acid comprise antisense and RNA (for example siRNAs) and vaccine.

Of the present invention one preferred aspect, compositions of the present invention makes when being exposed to aqueous fluid and to form I ₂Or L ₂Phase or its mixture, and in compositions, comprise the polarity activating agent.Suitable especially polarity activating agent comprises peptide and protein active thing, comprises following those that list.The special in this regard peptide class Sandostatin LAR Depot peptide relevant meaningfully with other somatostatin, polarity activity chlorhexidine (for example chlorhexidine digluconate or chlorhexidine dihydrochloride) and diphosphate (for example her this Alendronate, Zoledronate (zoledronate), fosamax, Sodium Pamidronate, Disodium tiludronate etc.).

The one type of suitable especially activating agent that is included in any suitable aspect of the present invention is peptide/protein active thing.These comprise: hormone and hormone derivative be growth hormone for example; Somatostatin (and analog); Calcitonin (people or salmon); Oxytocin; Luetinizing hormone releasing factor is (with derivant leuprorelin acetate for example; Coserelin and triptorelin); Vassopresin (with derivant for example Desmopressin and felypressin); α and β follitropin; Human chorionic gonadotropin-β; Thyrotropin-α; Secretin (for example pig); Bradykinin; Hypotension tissue hormone; Insulin-α and insulin-β; Antiviral, antibiotic and anti-fungus peptide class comprise interferon-' alpha ' 1/13, interferon-' alpha ' 2, interferon-beta, interferon-(comprising reorganization shape), king crab Copeptin (tachyplesin) i, stimulin (tuftsin), magainin i and ii, unfolding peptides (indolicidin) (for example cattle), protegrin (for example pig), polyphemusin i and ii, polymyxin b, Gramicidin s; Interleukin (ils) comprises il-1 α; Cytagenin (hematopoietin)-1; Il-1 β; Catabolin; Il-2; T-cell growth factor (tcgf) (recombination leukocyte mesonium-2); Il-3; Hematopoietic cell growth factor; Il-4; The b-LSF; Il-5; The t-cell replacement factor; Il-6; The b-LSF; Il-7; Il-8; The neutrophil activation agent; Il-9; T-cell growth factor p40; Il-10; CSIF; Il-11; Adipogenesis inhibitory factor; Il-13; Il-15; Il-17; Cytotoxic t lymphocyte-associated antigen 8; Il-18; The interferon gamma inducible factor; Il-19; Melanoma differentiation related protein appearance protein; Il-20; Four α spiral cell factor zcyto10; Il-24; Melanoma differentiation related protein 7; Il-26; And other peptide and protein, comprise ICAIU 1, pneumadin, alteplase, interleukin-1 receptor antagonist, gmcsf, filgrastim (g-csf), lepirudin, becaplermin, ospa, avicine, tubulysins a-f, contakulin g (cgx-1160), α conotoxin appearance peptide (seeing WO 02/079236) and metformin.

In Therapeutic Method of the present invention, and in corresponding treatment purposes and drug manufacture, activating agent is always unessential.Especially, lipid, particularly phospholipid for example PC be implied to be itself the treatment of some disease (comprise following those) be highly profitable.Not by any theory constraint, those in the lipid of the thinking fit preparation for example of the present invention are on the contact surface in the lining form (linings) of many structure example of health such as many body cavitys and joint and form protective layer on every side.These layers can be used for protecting the adhesion that avoids number of chemical agent and biological agent and attack (for example on stomach surface with the lining form in GI road); Can play the effect (especially on joint and most important ground also in many internal structures for example on the lining form around heart and the lung and the film) of lubricant, and other dilution through permission lipid exchange and unwanted film bonding agent and film lytic agent can help the reparation of cell wall.The lipid in nature of compositions also forms unwanted inflammatory lipase for example phospholipase such as phospholipase A ₂(PLA ₂) harmless substrate.

In the alternate embodiment of Therapeutic Method of the present invention and corresponding uses, it is involved that suitable active matter can be used as single beneficial agent, or in order to replenish the effect of suitable lipid composition.Suitable active matter will typically be applicable to treatment inflammation and/or stimulation, for example steroidal and NSAID and local immunomodulator.The instance of such reagent is known and comprises corticosteroid for example prednisone, Methyllprednisolone and hydrocortisone, and the derivant of nonsteroidal anti-inflammatory compound for example benzydamine, acetaminophen, ibuprofen and salicyclic acid derivatives comprise acetylsalicylate and 5-aminosalicylate.The local inhibitor of inflammation passage also is suitable for, and comprises for example PLA of antigen recognition inhibitor methotrexate, azathioprine or Ismipur and inhibitor of phospholipase enzymes ₂Inhibitor.In context, it should be noted that compositions of the present invention is applicable to that intraarticular is administered in the synovial fluid, wherein phospholipid has known beneficial effect about joint lubrication except being the controlled release carrier.

Through with reference to its known dose, consider route of administration, with the suitable load of confirming activating agent, and to compare than known formulations, compositions of the present invention can provide the bio-absorbable of bigger activating agent.

A particularly advantageous aspect of compositions of the present invention is to introduce very high-caliber activating agent.Particularly, the compositions that comprises certain proportion water or cosolvent (described herein) is very effective in the high-caliber polytype activating agent of dissolving.Therefore, compositions can comprise at least 2% activating agent, preferred at least 5% and more preferably at least 10% activating agent.Advantageously, can introduce the activating agent of as many as 20wt%.

The particle (comprise by maybe can forming of forming of the present composition those) that the present invention is based on amphiphilic substance also can needs with surfactant (especially polymer) modification, for example starch or starch derivatives, the copolymer (for example ethylene oxide/propylene oxide block copolymer) that contains alkylene oxide residue, cellulose derivative (for example hydroxypropyl emthylcellulose, hydroxyethyl-cellulose, ethylhydroxyethylcellulose, carboxymethyl cellulose etc.) or its grafted hydrophobically modified derivant, arabic gum, hydrophobically modified polyacrylic acid or polyacrylate etc.Surface-active polymer also can have functional impact to particle surface, for example, and for the combination of particle selection property or be targeted to its ideal action site.Particularly, for example polyacrylic acid, hyaluronic acid, gellan gum or chitosan can be used for providing the mucosa-adherent particle to polymer.Therefore, such particle will trend towards keeping localization, therefore increase the spatial control that activating agent is discharged.The present composition that contains such surface-modified particles forms another embodiment of the invention.

In colloidal composition, particle mean size typically in 0.1 to 0.6 mu m range, is for example measured through light scattering method (for example laser diffraction).Preferably, being no more than 1% particle will be beyond the scope of 0.05 to 1.5 μ m, and more preferably, being no more than 0.1% particle will be at this beyond scope, but and most preferably beyond this scope, does not have a particle (through laser diffraction) of detection ratio.In non-colloidal state preparation, particle mean size will be typically in 10 to 200 mu m ranges.

In addition, the advantage of a highly significant of the present invention is: the colloidal state preparation typically is a physically stable for long-term storage under the ambient temperature.Such preparation will at room temperature at least 10 days, more typically at least 3 months, preferred at least 6 months and more preferably in period of 12 months or longer time, with regard to phase behavior, granularity and particle size distribution, are stable basically.By contrast, the dispersion of known similar granularity at room temperature particle size stable be less than 10 days (seeing for example Kamo etal supra).This is the special benefits that the present invention contains the compositions of component a+b+c, and this is to lack the compositions of amount of component b because containing component a+b for storing usually instability.

If be no more than twice, can think that particle size distribution is stable basically for storage in the increase of storage period particle mean size.Preferably, should be no more than 50% in the increase of storage period average-size, more preferably no more than 20%.Similarly, at storage period, the dispersion of distribution should preferably increase and is no more than 50% when half-peak was high, more preferably no more than 20%, was most preferably not exceeding 10%.Be distributed as under the unimodal situation, should preferably remaining unimodal at storage period.In an especially preferred embodiment, the change of particle size distribution width is no more than 10% and remain unimodal when the particle mean size of the particle size distribution of the above-mentioned storage period present composition and half-peak are high.

When colloidal dispersion was used for intravenous or intra-arterial and uses, particle size distribution is stable between storage and operating period to be particular importance.Contain even the compositions of few relatively non-colloidal particle component can cause thromboembolism, or cause the rate of release that to expect in being applied directly to blood flow the time at least.Similarly, the controlled release of activating agent can be depending on the reliable particle size distribution in the compositions of using through any other approach.Hope that medicine, diagnosis article and article for animals also are stable or the cost and the obtaining property of said product affect adversely significantly in the storage of several months.

In the other important and particularly preferred embodiment of the present invention, fluid composition of the present invention can be prepared into solvent mixture.Such Liquid precursor will comprise component a, b, c, cosolvent and optional activating agent.For example, the said Liquid precursor that comprises activating agent can be filled in the capsule, and when contacting with the GI-fluid, forms non-booklike particle.Similarly, the injection before can with fluid precursor be fed to be used for being scattered in fluid (for example isotonic saline solution) but ampoule or direct injection and when contacting, form non-booklike particle in vivo with body fluid.The most important thing is that the inventor finds through changing the content of amount of component b unexpectedly, possibly adjust release duration window in the body several weeks from several hours to as many as.In addition, high initial drug concentration be can avoid, potential part and systemic side effects therefore reduced.

General cosolvent should be can be with water miscible or part is water-soluble at least, and should in the application that will use compositions, can tolerate.Has 1 to 6 carbon atom and preferably the substituent organic solvent of at least one oxygen atom and water-soluble polymer thereof are preferred.Suitable cosolvent kind is alcohol (comprising polyhydric alcohol), ketone, ester, ether and polymer thereof.Typical cosolvent is ethanol, isopropyl alcohol, N-N-methyl-2-2-pyrrolidone N-(NMP), propylene glycol, PEG400 and glycerol.Ethanol is particularly suitable.Can solvent be added to the level of the highest about 10 to 20% (by weight) of TL.

Dispersion through preparation component a, b and c in solvent (for example aqueous solvent) can form compositions of the present invention, adopts one or more heating and cooling circulations randomly to handle dispersion then.

Before optional heat treated circulation, the particle dispersion that will comprise component a, b and c forms preformulation.Through the method for confirming; The for example embodiment of the invention and US 5; 531,925, those that describe among WO02/02716, WO 02/068561, WO 02/066014 and the WO 02/068562 can prepare this preformulation, and this preformulation itself can be a compositions of the present invention.These disclosure and all reference papers of here quoting are at this by reference as a reference.

These methods comprise:

I) (for example the component in water a) is added in the aqueous solution of cracking agent (for example components b and/or c), or makes that mixture is broken naturally or adopt that for example mechanical agitation, vortex, rotor stator (roto-stator) mixing, high pressure homogenizing, Micro Fluid (microfluidisation) and/or ultrasonic the acceleration are handled with amphiphilic substance/water liquid crystalline phase; Or

Ii) the mixture (randomly comprising at least a bioactivator) with a+b+c adds in the solvent (for example aqueous solution) and directly stirring.

Can prepare another method that (especially from liquid crystalline phase) contain the dispersion of activating agent is through being dissolved in supercritical carbon dioxide (sc-CO ₂) or be suitable for dissolving and reducing the alternate processing solvent of conduct of composition viscosity, for example in the lower alcohol (for example methanol or ethanol).Especially, the liquid crystalline phase of body cube or hexagonal phase for example, normally high viscosity and possibly be difficult to handle and mix.Therefore, if liquid crystalline phase is prepared into bulk liquid and supported active agent subsequently, is difficult to realization so and need makes the equally distributed mixing of activating agent.At the supercritical region of Pressure/Temperature phasor (typically under room temperature or above and 150 crust or bigger pressure), carbon dioxide forms very effective solvent and can be used for reducing the viscosity of liquid crystalline phase, and promotion mixes and load with the effective of activating agent.Then, can remove said sc-CO ₂(for example through reduce pressure), and this bulk phase of load is dispersed in the solvent, as stated.Therefore, use sc-CO in the dispersion liquid crystalline phase that forms the activating agent load (especially of the present invention those) ₂Constitute another aspect of the present invention.

Can be through the phase behavior and the distribution of sizes of one or more (preferred one) heating and cooling loop control particle preparation of the present invention.Such circulation can be used for booklike particle is changed into non-stratiform form, and/or is used to reduce particle size distribution.The stability of particle also can be improved through the method.

Heat cycles is elevated to the compositions that has or do not exist activating agent is enough to feasible part particle at least changes into non-lamellar phase when being cooled to ambient temperature temperature.This will typically comprise be heated to about 90 to 150 ℃ 1 to 30 minute, be cooled to ambient temperature subsequently.More typical heat cycles will be included in the cooling before be heated to about 100 to 120 ℃ 2 to 20 minutes.Optimal condition will detail change between the different combinations thing, but those skilled in the art can easily confirm.

In heat cycles was handled, particle mean size increased and the particle size distribution reduction usually slightly.

On the other hand, therefore the present invention also is provided for forming the method for non-booklike particle, and said method comprises the formation mixture, and said mixture comprises:

A) 5 to 90% at least a phosphatidylcholine component,

B) 5 to 90% at least a DG component, at least a tocopherol or its mixture and

C) 2 to 40% at least a nonionic stability amphiphilic substance,

Wherein all parts and are dispersed in said mixture in the aqueous fluid with respect to a+b+c weight summation by weight.This method preferably also comprises at least one heating and cooling as described herein circulation, aqueous solution, body fluid or any other suitable fluid as described herein that aqueous fluid can be water, be suitable for injecting.Other can is made up of or also can comprise to mixture fully amphiphilic substance a-c component is activating agent and/or the mixable solvent of water for example, like following embodiment illustrated.This method also randomly has drying steps (for example spray drying or lyophilization) subsequently, thereby compositions is formed powder type.

With preferably from a series of freezing-the transmission electron microscope particle picture estimates the existence of non-stratiform form particle, preferably shows more than 20, preferred more than 30 and the most preferably sample of at least 50 particles.Also can be through the existence of the non-booklike particle of X-ray scattering test evaluation.

Because heating treatment method can be used for booklike particle is changed into non-stratiform form, thus the preformulation particle unnecessary be unstratified.Therefore, any method of knowing that lipid is mixed with vesicle can be used for producing the preformulation that uses in the heating treatment method of the present invention.Appropriate method comprises, and is for example, ultrasonic or extrude (for example passing through polycarbonate membrane).Such method will be that the technical staff in the suitable field knows.

Preferably, should prepare preformulation and make that the Thermodynamically stable state is unstratified at ambient temperature, this generally will be because component a, b and the concrete selection of c and the situation that ratio causes thereof.Alternatively, said non-stratiform form can be the thermodynamics metastable condition.When having activating agent, before the heat cycles and/or afterwards, can said activating agent be incorporated in the particle.When using an above heat cycles, can between circulation, introduce activating agent.Under the heat sensitive situation of activating agent (for example peptide or protein), preferably, heat cycles introduces activating agent after accomplishing.By contrast, under the activating agent situation stable to the heat cycles method, the method (heat cycles that has activating agent) can be used for providing very high activating agent load, its stable for extended periods of time.

Particle (heat treated or heat treated subsequently) can concentrate (for example ultrafiltration or dialysis) and/or drying, for example through spray drying, fluid bed drying or lyophilization.Under the situation of drying particulate, dried can be through single or multiple gathering and granulation step expansion granularity subsequently.Spissated, the exsiccant and/or accumulative particle preparation that so forms can so use, or hydration and/or disperse to obtain being applicable to the non-booklike particle dispersion of delivery of active substances (especially in vivo).Spissated, exsiccant and/or accumulative particle preparation like this and the dispersion that is obtained by its resuspending/hydration constitute another aspect of the present invention.

But resuspending drying of the present invention (it refers on the function dry rather than does not contain solvent fully) powder composition is suitably to obtain colloidal state or non-colloidal dispersion in (especially aqueous) fluid.Alternatively, said dry compsns may be dissolved in the suitable cosolvent as described herein and uses, thereby when contacting with body fluid, forms non-layer structure in vivo.These aspects of the present invention are particularly suited for intramuscular and/or subcutaneous injection and can form persistent non-layer structure, and activating agent can slowly release from said persistent non-layer structure in the period of a couple of days or several weeks.Such slow releasing preparation can result from any suitable compositions of the present invention, but is particularly suited for from the powder of resuspending, producing.

Through in compositions of the present invention, using polymeric reagent can prepare semisolid (example gel, waxy solid) compositions.Such semi-solid precursors will comprise compositions of the present invention as described herein and other at least a polymer coagulant.Typically, such compositions comprises component a, b, c, polymeric reagent, optional cosolvent and optional activating agent.Said semi-solid precursors normally can be through heating liquefaction, and for example can be filled in the capsule, molded etc.But resuspending semi-solid combination of the present invention is suitably to obtain colloidal state or the non-booklike particle dispersion of non-colloidal state in (especially aqueous) fluid.Alternatively, when for example the GI-fluid contacts with aqueous body fluid, form non-layer structure.The preferably biological polymer that can tolerate of polymer coagulant, preferred fusing point is at 35 to 100 ℃, more preferably at 40 to 95 ℃, most preferably at 45 to 90 ℃.Preferred especially polymeric reagent is Polyethylene Glycol (PEG), its molal weight in 950 to 35000 scopes, most preferably 1000 to 10,000.PEG4000 is special preferred examples.

Preparation of the present invention comprises at least a compositions of the present invention and at least a appropriate carriers or excipient.At preparation is under the situation of pharmaceutical preparation, and said carrier or excipient will be can tolerate on the medicine.

Adopt conventional pharmaceutical carrier, diluent and/or excipient for example aqueous carrier (for example water for injection), binding agent, filler, stabilizing agent, osmotic pressure regulator, antioxidant, foaming agent, pH buffer agent and regulator, viscosity modifier, sweeting agent, lubricant, emulsifying agent, flavoring agent, coating materials (for example resisting the coating of gastric juice) but etc. compositions formulated.

The preparation that comprises the present composition and at least a medicine acceptable carrier and/or diluent can be mixed with any known dosage form, comprises as the suspensoid in the liquid, powder, tablet, capsule, coating capsule (coated capsules), coated tablet, aerosol, suppository, drop, cream, transdermal patch, spray etc.Under the exsiccant situation of compositions of the present invention, before using, this can be with suitable form (for example powder) preparation, with resuspending in suitable medium (for example purified water or physiology isosmotic solution).Can comprise oral, eye usefulness, suction, parenteral (for example passing through intramuscular, subcutaneous or intravenous injection or infusion), part, rectum etc. through any appropriate method, come administered formulation and pharmaceutical composition.The parenteral compositions is preferred, and this is because the present invention provides high stability (aspect granularity and phase structure) and the toxic significant combination of low-down parenteral.Yet topical compositions also is very effective, and the spraying dispersion of pump spraying (pump-spray) dispersion or pressurization can be used for skin, nasal cavity and mouthful interior (an especially buccal (buccal)) application.Rectal administration as spissated dispersion or solid-state suppository also is fit closely.

The preparation of the present invention, compositions and the method that relate to inflammation or stimulation therapy are particularly suitable for solving inflammation and/or stimulation in the body cavity.Therefore, in this regard, in body cavity, use and be fit closely and will carry out through being suitable for waiting the method for treating the chamber.Collutory for example, can be suitable for oral or oral cavity, but and other part administered through oral preparation and the rectal formulation in GI road are suitably treated, said oral formulations comprises dispersion and exsiccant preformulation, said rectal formulation for example is enema or suppository.Lotion (Rinses) and pesseries are suitable for vagina similarly and send.

Compositions of the present invention is very suitable for treating inflammation in the body cavity, and this is because the height bioadhesion character of non-lamellar phase and the persistent effect of gained.Preparation comprises or the intrinsic comfort property and the biocompatibility of height that are dispersed into ability and the component of non-booklike particle also is important, said then non-booklike particle easily transmit and be distributed in application site around.

Therefore, Therapeutic Method of the present invention and corresponding use are best suited for inflammatory diseases and the inflammation that is for example caused by wound or scratch.What especially be fit to is the inflammatory diseases that infects at least one body cavity.The disease in GI road is fit to adopt compositions of the present invention to treat very much, and particularly inflammatory bowel disease comprises the sick and ulcer property collitus of Crohn ' s.Similarly, also can use and be applied to body cavity, to utilize the characteristic of preparation at intra-operative.Therefore, for example through spraying or apply can they directly applied to alleviate and come from or inflammation that intra-operative exposes, also be used to reduce " adhesion " of surgical procedure tissue and/or in the tendency of undesirable position formation adhesion/bridging.

The special advantage of the present composition is their significantly low toxicity, especially when parenteral is used.Particularly, when using through intravenous injection, compositions of the present invention can show significantly low acute toxicity.One preferred aspect, therefore the present invention provides with 200mg/kg body weight at least, preferred 600mg/kg at least when more preferably rat is given in the horizontal intravenous injection of 1g/kg body weight at least, the present composition of the no acute toxicity of demonstration.

As stated, another key advantage of the present composition is that they can be used for producing " short-term " depot compositions.Especially, the quick releasing formulation of activating agent is common, and coating etc. can be used for being provided at the preparation that is up to release bioactive agent in about 12 hours period sometimes.By contrast, long-acting " depot " injection typically comprises the polymer for example solution or the suspension of polylactic acid-common-hydroxyacetic acid, or passes through the physics pump of the implantation health of osmotic pressure driving.These methods typically provided one month or the release of above time, and typically needed complicated preparation and/or use.By contrast, seldom method can be as required in 1 to 30 day period during for example operation back analgesia or the antibiotic, especially in 1 to 14 day time, and release bioactive agent in 2 to 7 day time most preferably.Compositions of the present invention has or the more how following advantageous feature that provides with its specific composition.

They provide seldom what need or need not prepare is the injection-type compositions;

They have been avoided the tediously long preparation that the depot product typically needs and have used;

They can be directly by the preparatory filling injection device injection (it also constitutes one aspect of the present invention) that contains compositions;

Their storage-stables, as stated;

They can pass through pore pin (for example less than No. 20, preferred No. 23 or littler, more preferably No. 27 or littler) injection;

They are intramuscular or subcutaneous or intracavitary administration effectively;

Can introduce the activating agent (as described herein) of high-load level;

Particularly advantageous short-term bank is the sort of (preferred dose 0.1 to 20mg) that comprises the present composition and GLP-1 or its analog or derivant, is used for subcutaneous or intramuscular injection.Such bank can provide the GLP-1 analog in 2 to 14 day time, the lasting release in preferred 5 to 10 day time.The optimal purposes of such compositions can be treatment diabetes (especially II type) or produce the medicine that is used for this purposes.

Glucagon-like peptide (GLP-1) is that strong glucose of imitating is regulated hormone, its by small intestinal L cellular response in nutrient picked-up and neural with the endocrine stimulation and be discharged in the circulation.On the structure, GLP-1 is that MW is 37 amino acid whose peptides of 4.2KDa, has the sequence of high conservative between variety classes.GLP-1 relates to insulin secretion and the biosynthesis that strengthens the glucose stimulation through comprising, and the effect of glucagon suppression secretion, gastric emptying and food intake adjustment glucose homeostasis.The ability that GLP-1 stimulates insulin secretion and glucagon suppression discharges is that glucose is dependent; Therefore, the risk of hypoglycemia used of GLP-1 is low.GLP-1 also stimulates beta-cell proliferation and regeneration and suppresses the beta cell mechanism of apoptosis through comprising, increases the beta cell amount in the diabetes preclinical models.Research among the animal and human shows that GLP-1 also can play protective effect in the cardiovascular system.

The synergy of GLP-1 is to having produced substantive meaning with this peptide as the therapeutic agent of treating type 2 diabetes mellitus.Yet the treatment potential of natural GLP-1 receives its very restriction of short plasma half-life (being lower than 2 minutes).This since proteolytic enzyme dipeptidyl peptidase (DPP)-IV and kidney removing cause.Therefore; GLP-1 analog long-acting, the DPP-IV-tolerance is used for clinical use by exploitation; Comprise exenatide (Byetta, Amylin-Lilly), liraglutide (NovoNordisk), CJC-1131 (ConjuChem), AVE010 (ZealandPharma-Sanofi-Aventis), LY548806 (Lilly) and TH-0318 (TheraTechnologies).All these be once a day or every day administered twice product; Controlled release (1 week) exenatide product (Alkermes-Amylin-Lilly) is in the clinical research at present.These GLP-1 analogies are combining the GLP-1 receptor and to produce those the biological agent be equal to natural GLP-1 with the similar affinity of natural GLP-1, but the inactivation of tolerance DPP-IV-mediation and kidney removing.These chemical compounds play more persistent GLP-1-appearance activity in the longer time in vivo.The replacement therapy method that prolongs natural GLP-1 effect is to suppress the DPP-IV activity, thereby prevents the GLP-1 degraded.Estimate several kinds of active Peroral active agents of inhibition DPP-IV and be used to treat type 2 diabetes mellitus.

One other aspect; The present invention is provided for the test kit with the suspensoid prepare present composition; Said test kit comprises the compositions of the present invention of at least a powder type and randomly with preferably with the description of powder suspension in aqueous fluid.

Description of drawings

Referring now to following indefiniteness embodiment and accompanying drawing, further illustrate the present invention, wherein:

Fig. 1 shows before the heat treated and the particle size distribution of dispersive non-stratiform SPC/GDO/P80 sample afterwards.

Fig. 2 shows heat treated before with afterwards, the particle size distribution of dispersive non-stratiform SPC/GDO/P80 sample.

Fig. 3 shows heat treated before with afterwards, the freezing-transmission electron microscope photo of dispersive non-stratiform SPC/GDO/P80 sample.

Fig. 4 shows heat treated before with afterwards, the particle size distribution of dispersive non-stratiform SPC/GDO/Solutol HS 15 samples.

Fig. 5 shows after the heat treated, freezing-transmission electron microscope photo of dispersive SPC/GDO/Solutol

HS 15.

Fig. 6 shows heat treated before with afterwards, the particle size distribution of the non-stratiform SPC/GDO/P80 sample of spissated dispersion.

Fig. 7 shows preparation back and at 25 ℃ of storages after 2 months, the particle size distribution of dispersive non-stratiform SPC/GDO/P80 sample.

Fig. 8 shows preparation back and at 25 ℃ of storages after 2 months, the particle size distribution of dispersive non-stratiform SPC/GDO/P80 sample.

Fig. 9 is presented at the particle size distribution of two different propofol-amphiphilic substances than the non-stratiform SPC/GDO/P80 particle dispersion of the dispersion of load anesthetic propofol.

Figure 10 is presented at the PC of using the back propofol in the rat medium-sized vein.

Figure 11 shows the particle size distribution of dispersive non-stratiform SPC/ alpha-tocopherol/vitamin E TPGS sample.

Figure 12 shows the particle size distribution of dispersive non-stratiform EPC/GDO/P80 sample.

Figure 13 shows heat treated before with afterwards, freezing-transmission electron microscope photo of dispersive non-stratiform sample SPC/GDO/P80.

Figure 14 shows heat treated before with afterwards, freezing-transmission electron microscope photo of dispersive non-stratiform sample SPC/GDO/P80.

Figure 15 is presented in the rat PC of Sandostatin LAR Depot after the subcutaneous administration.

Figure 16 is presented in the rat PC of Sandostatin LAR Depot after the subcutaneous administration.

Abbreviation

The SPC=S-PC derives from Lipoid GmbH, Germany

The GDO=NOFABLE GO-902P derives from Danisco, Denmark

P80=Polysorbate 80, derive from Apoteket, Sweden

Solutol HS 15=Polyethylene Glycol 15 hydroxy stearic acid esters; Derive from BASF, Germany

Freezing-TEM=is freezing-transmission electron microscope

The PPF=propofol derives from Sigma-Aldrich, Sweden

The EPC=Yolk lecithin derives from Lipoid GmbH, Germany

DOPE-PEG (5000)=dioleoyl PHOSPHATIDYL ETHANOLAMINE Polyethylene Glycol 5000 derives from AvantiPolar Lipids, U.S.A.

CMC=carboxymethyl cellulose (sodium salt) derives from Sigma-Aldrich, Sweden

The PVP=polyvinylpyrrolidone derives from ISP, U.S.A.

The PEG=Polyethylene Glycol derives from Merck, U.S.A.

The non-stratiform anti-phase of embodiment 1-nanoparticle

1.1-the preparation of non-lamellar dispersions

Through mix 2.125g SPC/GDO40/60wt/wt mixture (through in ethanol, mix lipid and subsequently evaporating solvent form) and 0.3826g P80 form non-stratiform (amphiphilic substance＞80wt%) and stratiform (and amphiphilic substance＜20wt%) dispersion of nano-particles body.Through mixing said component at molecular level in 70 ℃ of heating 5 minutes and vortex.Fused even lipid (2.012g) is dropwise joined in the 38.01g deionized water.The coarse dispersion of gained was placed on the jolting platform (350rpm) and jolting 24 hours, to obtain muddy homogeneous dispersion.

Adopt laser diffraction (Coulter LS230) to measure granularity.Find that distribution of sizes is narrow with unimodal, particle mean size is 95nm.

1.2-heat treated

The heat treated of the enterprising line option of dispersion of preparation circulation in embodiment 1.1.

With dispersion samples (25mL) autoclaving (125 ℃, 20 minutes) that produces among the embodiment 1.1 and be cooled to room temperature.Particle size distribution is narrow, and particle mean size is increased to 137nm, freezing when adopting-during tem analysis, still more the particle of vast scale shows non-stratiform characteristic.Fig. 1 shows before the heat treated and particle size distribution afterwards.

Component:

a SPC

b GDO

c P80

Preparation

a∶b∶c

abc wt％

Medium

Water wt%

Mutually

Temperature ℃

Time minute

Mutually

i

33.9∶50.8∶ 15.3

5.0

Deionized water

95

Non-stratiform

125

20

Non-stratiform

Other compositions of embodiment 2-

Method through embodiment 1.1 and 1.2 prepares second compositions, estimates the influence of adding the higher concentration stabilizing agent.Through coming at 70 ℃ of heating 5 minutes and vortex (2.017g) and the solution of P80 (0.514g) at molecular level mixing SPC and GDO (40/60wt/wt).Fused even lipid (2.006g) dropwise is added in the 38.00g deionized water.Place on the jolting platform coarse dispersion of gained and jolting 24 hours, to obtain muddy homogeneous dispersion.Thereafter according to embodiment 1.2 heat treated dispersions.

Find before the heat treated with afterwards distribution of sizes be narrow with unimodal, particle mean size is respectively 88 and 129nm.As shown in Figure 2, heat treated also makes size distribution narrow.Fig. 3 shows before the heat treated and the freezing-TEM photo that obtains from sample afterwards.Freezing-TEM result clearly confirms the formation of the non-laminar nano grain of consistent size, and the non-laminar nano particle of wherein said consistent size comprises unordered many places and connects the double-layer inner structure.Find that particle shows than those the finer and close nuclears before the heat treated after heat treated.

Component:

a SPC

b GDO

c P80

Preparation

a∶b∶c

abc wt％

Medium

Water wt%

Mutually

Temperature ℃

Time minute

Mutually

ii

31.9∶47.8∶ 20.3

5.0

Deionized water

95

Non-stratiform

125

20

Non-stratiform

This concrete compositions also is suitable for preparing the Liquid precursor of non-lamellar phase dispersion very much.Identical component is used with identical ratio.Through mixing said component at molecular level in 70 ℃ of heating 5 minutes and vortex.Adopt the cosolvent (for example ethanol, N-N-methyl-2-2-pyrrolidone N-(NMP), propylene glycol, PEG400, glycerol) of 10wt% also can strengthen Liquid precursor; Thereafter the mode with gentle jolting is distributed in the water (5wt% amphiphilic substance), obtains the milky dispersion of non-lamellar phase particle.

Other compositions of embodiment 3-

Method through embodiment 1.1 and 1.2 prepares another kind of compositions, estimates the influence of the stabilizing agent that adds another kind of type.Through coming at 70 ℃ of heating 5 minutes and vortex (2.004g) and the solution of Solutol

HS 15 (0.516g) at molecular level mixing SPC and GDO (40/60wt/wt).Fused even lipid (2.042g) is dropwise joined in the 38.00g deionized water.Place on the jolting platform coarse dispersion of gained and jolting 24 hours, to obtain containing the muddy dispersion of dispersive macroparticle a little less than some.In order to obtain homogeneous dispersion, adopt the Microfluidizer sample that under 5000PSI and room temperature, homogenizes.With sample through homogenizer 5 times to obtain the emulsus homogeneous dispersion.Thereafter according to embodiment 1.2 heat treated dispersions.

Fig. 4 shows before the heat treated and the distribution of sizes that obtains afterwards, shows that heat treatment step obtains unimodal and narrow distribution, and particle mean size is 343nm.Freezing-TEM test after the heat treated shows the particle with fine and close non-layer structure in inside, and is as shown in Figure 5.

Component:

a SPC

b GDO

c Solutol

HS?15

Preparation

a∶b∶c

abc wt％

Medium

Water wt%

Mutually

Temperature ℃

Time minute

Mutually

iii

31.8∶47.7∶20. 5

5

Deionized water

95

Non-stratiform

125

20

Non-stratiform

Other compositions of embodiment 4-: spissated non-booklike particle dispersion

Method through embodiment 1.1 and 1.2 prepares spissated non-booklike particle dispersion.Through coming at 70 ℃ of heating 5 minutes and vortex (4.7958g) and the solution of P80 (0.8152g) at molecular level mixing SPC and GDO (40/60wt/wt).Fused even lipid (5.001g) is dropwise joined in the 44.999g deionized water.Place on the jolting platform coarse dispersion of gained and jolting 48 hours (350rpm), to obtain muddy homogeneous dispersion.Thereafter according to embodiment 1.2 heat treated dispersions.

Find before the heat treated with afterwards distribution of sizes be narrow with unimodal, particle mean size is respectively 103 and 174nm.As shown in Figure 6.

Component:

a SPC

b GDO

c P80

Preparation

a∶∶b∶c

abc wt％

Medium

Water wt%

Mutually

Temperature ℃

Time minute

Mutually

iv

34.2∶51.3∶14.5

10

Deionized water

90

Non-stratiform

125

20

Non-stratiform

Embodiment 5-storage stability

Method according to embodiment 1.1 and 1.2 prepares non-lamellar dispersions.The composition that shows dispersion in the following table.Dispersion is stored at 25 ℃ and with the interval measurement particle size distribution of rule.It is consistent to find that at least 2 months distribution of sizes of storage and original size distribute, and this shows fabulous colloidal state and storage stability.

Can be observed at storage period (through freezing-TEM) not variation of form of non-booklike particle.Fig. 7 (SPC/GDO/P80=34/51/15wt%) and 8 (SPC/GDO/P80=32/48/20wt%) show raw dispersion and the particle size distribution of storage after 2 months.As observable, original distribution of sizes with the dispersion of storing is identical basically.

Be used to study the composition table of storage stability

Compositions	Weight ratio	Lipid concentration (wt%)	Medium
				SPC/GDO/P80	34∶51∶15	5	Deionized water
SPC/GDO/P80	32∶48∶20	5	Deionized water

The load of embodiment 6-activating agent

Contain SPC (32wt% amphiphilic substance), GDO (48wt% amphiphilic substance) and P80 (20wt% amphiphilic substance) and PPF through the mixed shown in the following table, form the non-booklike particle dispersion that contains narcotic activity agent PPF.Through at 70 ℃ of heating 5 minutes and vortex, come at the molecular level blending ingredients.Fused even lipid/PPF is dropwise joined in the aqueous solution that contains 2.5% (in the weight of total preparation) glycerol.Place jolting platform (350rpm) to go up also jolting 12 hours the coarse dispersion of gained, to obtain homogeneous dispersion.Thereafter according to the method heat treated dispersion of embodiment 1.2.Dispersions obtained particle size distribution is narrow with unimodal, and particle mean size is 140 in the 150nm scope, and is as shown in Figure 9.The dispersion of finding the PPF load was a storage-stable at room temperature at least 2 months.

The composition table of final non-booklike particle/PPF dispersion

Amphiphilic species concentration (mg/mL)	PPF concentration (mg/mL)	PPF: amphiphilic species (wt: wt)
			50	10	1∶5
50	20	1∶2.5

Embodiment 7-is carried on the pharmacokinetics and the drug influence kinetics of the propofol of non-booklike particle

Except PPF concentration in the case is that 10mg/mL and amphiphilic substance concentration are 25mg/mL (PPF: amphiphilic substance=1: 2.5wt/wt), adopt composition identical among the embodiment 6 and identical method preparation to comprise the dispersion of the non-booklike particle of PPF.With non-booklike particle PPF dispersion at rat (male SPF Sprague-Dawley rat (Mo1:SPRD HAN; M&B Taconic; Lille Skensved, propofol Fresenius Kabi Emulsion (the 10mg PPF/mL) of the duration of anaesthesia in Denmark) and reference commerce usefulness compares.Dosage with the 10mgPPF/kg body weight carries out single intravenous push (after two kinds of situation injected, occurring induction of anesthesia immediately) to animal.For the drug influence kinetic parameter, write down recovery time (the real response time shown in attempting to stand firm).The result is summarized in the following table, shows that non-booklike particle PPF dispersion efficiently keeps required anesthetic action.

Drug influence kinetic parameter table

Preparation	The rat number of elements	Mean Time To Recovery (second) (standard deviation)
			Propofol Fresenius Kabi	5	377(89)
Non-booklike particle PPF dispersion	5	448(60)

Before giving dosage, (give dosage preceding 1 day), give 5 minutes, 15 minutes, 30 minutes, 1 hour, 3 hours, 6 hours and 24 hours blood sample collections (0.3mL) behind the dosage.Measure propofol in the rat plasma through the known HPLC of art technology scientist (HPLC) method.Propofol PC in time respectively with reference preparation and non-booklike particle propofol preparation similar (Figure 10).Calculate elimination half-life (t through non-chamber pharmacokinetics (PK) method _Half), the extrapolation PC (C when mean residence time (MRT), total body clearance (CL), 0 time ₀) and curve under the gross area (AUC _∞).Adopt trapezoidal rule with from C _LastBe extrapolated to the infinitely great AUC of calculating _∞

The pharmacokinetic parameter table

Preparation

n

t Half (hour)

MRT (hour)

CL(mL/h)

?C 0

?AUC ∞

Propofol Fresenius Kabi

4

0.90(0.50)

0.88(0.38)

4326(1507)

?2036(329)

?777(207)

Non-booklike particle PPF dispersion

5

1.97(0.85)

2.56(1.25)

2799(568)

?1187(501)

?1159(243)

P＜0.05 (t-check)

?

Not

Be

Not

Be

Be

Suppose that using propofol with non-stratiform propofol preparation will cause circulation time increase in the blood plasma.Observed PK parameter indicating has really realized increasing the situation that propofol exists.When analyzing MRT (unimolecule that is any medical compounds is trapped in the time in the circulation), this is the most tangible, and this compares with the reference product has increased about 3 times.The parameter of destiny in other reflection propofol body, the t of promptly non-booklike particle propofol preparation _HalfIncrease and CL reduction show that also propofol stops the longer time (can not check difference in statistics ground, but trend clearly) in blood plasma.And, the AUC of non-booklike particle propofol preparation _∞Increase, being illustrated under the equal dose said preparation, compare the time that is exposed to propofol with reference preparation longer.All observe the circulation time of supporting non-booklike particle propofol preparation can increase active component.

Embodiment 8-acute toxicity test

Adopt following component to prepare non-lamellar dispersions through the method for embodiment 1.1 and 1.2:

a)SPC

b)GDO

c)P80

The weight ratio of a: b: c=34: 51: 15, being dispersed in total amphiphilic substance concentration is in the water of 10wt%.In dispersion, add sodium chloride (NaCl) to reach 9mg NaCl/mL.Rat model medium-sized vein in injection after, test the acute toxicity of dispersion thereafter.

Non-lamellar dispersions shows: (do not have acute toxicity in the dose dependent research of 1g amphiphilic substance/kg) at dosage up to the 10wt% amphiphilic substance dispersion of 10mL/kg.

Sealing of embodiment 9-hydrophilic, water-soluble colorant

Be prepared as follows the non-booklike particle of sealing high water soluble coloring agent patent blue (Patent blue): according to embodiment 1 preparation 3.0g SPC/GDO/P80 (34/51/15wt%) preparation.In this solution, add 0.15g ethanol and mix said preparation through vortex mixed.In the 3.0gSPC/GDO/P80/EtOH preparation, add 0.20g patent blue (20mg/mL) aqueous solution.Mix the sample of gained through vortex mixed, to obtain even low viscosity preparation.In the 22.5g deionized water, add the 2.55g said preparation, and with the preparation of 350rpm jolting gained 18 hours to obtain an even blue sub prose style free from parallelism.After ultrafiltration (30000MWCO filter) dispersion; Deduct the absorbance of filtrating (not sealed part) with the absorbance (640nm) of raw dispersion; And with the absorbance measuring envelop rate (all absorbance measurings before, add TritonX100 (concentration in deionized water be 10wt%) to obtain clear solution) of difference divided by raw dispersion.

Find that envelop rate is 85%, this shows that non-booklike particle has the big potential of sealing the water-soluble active thing.

Sealing of embodiment 10-hydrophilic, water-soluble peptide

Be prepared as follows the non-booklike particle of sealing the water-soluble peptide Sandostatin LAR Depot: according to embodiment 1 preparation 1.0g SPC/GDO/P80 (34/51/15wt%) preparation.In this solution, add 0.10g ethanol and mix said preparation through vortex mixed.Thereafter the sample that adds 0.054g Sandostatin LAR Depot (35.5mg/mL) aqueous solution and mix gained through vortex mixed is to obtain even low viscosity preparation.In 9.0g saline (9mg NaCl/mL), add the 1.0g said preparation, and in the preparation of 350rpm vibration gained 18 hours to obtain homogeneous dispersion (the about 100nm of particle mean size).Through the 2.5mL dispersion partly is collected in the independent bottle through Sephadex G25 (PD-10) post and with lipid part and free Sandostatin LAR Depot, seal the Sandostatin LAR Depot that separation is sealed the peptide from non-.After adding TritonX100, analyze the Sandostatin LAR Depot concentration in lipid part and the free Sandostatin LAR Depot part through HPLC.

Find that envelop rate is 71%, this shows that once more non-booklike particle has the big potential of sealing the water-soluble active thing.

Embodiment 11-is from the non-booklike particle of SPC/ tocopherol mixture

SPC, alpha-tocopherol and ethanol (27/63/10wt%) (1.34g) (0.30g) mix with d-alpha tocopherol cetomacrogol 1000 succinate (vitamin E TPGS).Through at 40 ℃ of heating 15 minutes and vortex, come at the molecular level biased sample.Fused even lipid (1.0g) is dropwise joined in the 19g deionized water.With the coarse dispersion of gained place on the jolting platform and jolting 20 hours (350rpm) to obtain muddy even non-booklike particle dispersion.

Find that distribution of sizes is narrow with unimodal, particle mean size is 128nm, and is shown in figure 11.

Thereafter according to embodiment 1.2 heat treated dispersions, and the heat treated sample freezing-TEM photo shows that non-booklike particle contains unordered surface texture and fine and close inner non-layer structure.

Embodiment 12-uses EPC as the SPC substitute

EPC (1.539g), GDO (2.302g), P80 (0.685g) are mixed with ethanol (0.501g).Rotate biased sample 3 hours through vortex mixed and end-to-end (end-over-end), obtain even clear liquid.In sterilized water (28.335g), add said liquid preparation (1.665g),, the coarse dispersion of gained was mixed 18 hours with 400rpm on the jolting platform.The dispersion that obtains is uniformly, finds that distribution of sizes is narrow with unimodal, and particle mean size is 114nm, and is shown in figure 12.

Freezing-TEM result shows the formation of the non-laminar nano particle of uniform-dimension, and said non-laminar nano particle contains many places and connects double-deck unordered surface texture and fine and close inner non-layer structure.

The robustness of embodiment 13-compositions

Form the influence that variation is easy to prepare to non-laminar nano grain in order to study lipid, preparation SPC/GDO ratio variation and P80 are than constant sample.Component is mixed with ethanol, is distributed to thereafter in the sterilized water, described in embodiment 13, except sample #1128 is vibrated 48 hours.Particle mean size and polydispersity index after particle mean size after the final composition of following table show sample, the jolting (400rpm) and polydispersity index (PI) and the heat treated (according to embodiment 1.2).

The composition and the tables of data of the sample of preparation among the embodiment 14

Sample ID	Form (wt%) SPC/GDO/P80/EtOH/ water	Mean size/nm (vibration)	PI a(vibration)	Mean size/nm (HT b)	PI a (HT b )
						#1128	2.125/2.125/0.75/0.56/94.44	110	0.24	141	0.17
#1129	1.91/2.34/0.75/0.56/94.44	114	0.24	136	0.18
						#1130	1.70/2.55/0.75/0.56/94.44	116	0.23	134	0.19
#1131	1.49/2.76/0.75/0.56/94.44	115	0.23	127	0.18

A) PI=polydispersity index is defined as the ratio between distribution of sizes standard deviation and the average-size; B) HT=heat treated

From above-mentioned table in the result displayed, about change SPC/GDO than the time particle average-size and distribution of sizes, non-laminar nano plastochondria system is very firm.Also observe heat treated and under all scenario, make size distribution narrow (lower PI).Figure 13 and Figure 14 show respectively before the heat treated and freezing-TEM photo of sample #1128 and #1131 afterwards, and show that non-booklike particle has the unordered surface texture that the many places of sealing the fine and close non-stratiform nuclear structure in inside connect bilayer.

The preparation of the non-booklike particle precursor of embodiment 14-liquid

Prepare the non-booklike particle precursor of liquid through mixing SPC (1.45g), GDO (2.15g), P80 (0.90g) with EtOH (0.50g), and end-to-end subsequently rotation mixed 5 hours, obtain all even limpid liquid.

Through in the 0.198g sterilized water, adding 2.0g aforesaid liquid formulation preparation second preparation, vortex mixed obtained limpid and uniform liquid in 1 minute subsequently.Following table shows the accurate composition of the non-booklike particle precursor of liquid.

The composition table of the non-booklike particle precursor of liquid

Sample	Form (wt%)	Outward appearance
			The non-booklike particle precursor 1 of liquid	SPC/GDO/P80/EtOH＝ 29/43/18/10	Clarification, equal even light yellow liquids
The non-booklike particle precursor 2 of liquid	SPC/GDO/P80/EtOH/H 2O＝ 26.4/39.1/16.4/9.1/9.0	Clarification, equal even light yellow liquids

The syringe that utilization has No. 27 pins is the dispersing liquid precursor easily, or for example utilizes pump sprayer atomizing of liquids precursor easily.

The pharmacokinetics of the non-booklike particle precursor of the liquid of embodiment 15-load Sandostatin LAR Depot (OCT) behind subcutaneous injection

Of embodiment 15, prepare the non-booklike particle precursor of liquid that contains Sandostatin LAR Depot through the SPC of mixed shown in the following table, GDO, P80, EtOH and OCT (every g preparation 0.6mg Sandostatin LAR Depot).Limpid to obtain, the uniform liquid preparation of sample through end-to-end rotation mixing gained.

The composition table that contains the non-booklike particle precursor of Sandostatin LAR Depot

Sample ID	Form (wt%) SPC/GDO/P80/EtOH/OCT	Outward appearance
			2022OCT-C	28.78/43.17/17.99/10.00/0.06	Clarification, equal even light yellow liquids
2022OCT-E	34.18/51.26/4.50/10.0/0.06	Clarification, equal even light yellow liquids

With the dose volume of the 1mL/kg that is equivalent to every kg body weight 0.6mg OCT, give rat through subcutaneous injection with the non-booklike particle precursor formulation of liquid.For 2022OCT-C; Before giving dosage, (give dosage preceding 1 day), give 10 minutes, 30 minutes, 1 hour, 3 hours, 6 hours, 24 hours and 48 hours blood sample collections (0.3mL) behind the dosage; For 2022OCT-E, before giving dosage, give 1 hour, 6 hours, 24 hours, 48 hours, 120 hours and 168 hours blood sample collections (0.3mL) behind the dosage.

Through the OCT content in all blood samples of competitive immunization experimental measurement.In brief, the antibody in the OCT competition solution that exists in OCT peptide that applies on the microplate and the plasma sample.Remove remaining antibody moiety in the solution, quantize to combine the part of immobilization peptide, the concentration of OCT is inversely proportional in signal that obtains and the sample.

The pharmacokinetics of the OCT that in the non-booklike particle precursor of liquid, prepares and the pharmacokinetics of the OCT in the saline solution compare.Result among Figure 15 shows that non-booklike particle precursor formulation provides the initial blood plasma level of low OCT (low " prominent releasing ") (with brinish contrast, C _MaxReduce about 15 times), and discharge that (or slow release) persistent period is up at least 48 hours and discharge (or slow release) persistent period for 2022OCT-E for 2022OCT-C and be up at least 168 hours (1 week).

Embodiment 16-is dispersed in the pharmacokinetics of the OCT that is carried on non-booklike particle behind subcutaneous injection in the saline

Through in vial, mixing SPC (0.918g), GDO (1.377g), P80 (0.405g) and EtOH (0.30g), prepare the liquid fatty stock solution subsequently through end-to-end rotation 15 hours.OCT (5mg) is dissolved in the sterilized water (0.095g), 2.2g lipid stores solution is added in the Sandostatin LAR Depot aqueous solution.The mixture of vortex gained becomes evenly up to sample.Lipid/Sandostatin LAR Depot mixture (1.85g) is added in the saline (18.15g), and on vibration table with the dispersion (0.2mg OCT/mL) of 400rpm mixing gained 15 hours.Thereafter through aseptic filtration (0.22 μ m filter) to the dispersion degerming.The dispersion of gained is muddy to milky and uniform, and like the about 100nm of particle mean size through laser diffraction measurement.

To be equivalent to the 3mL/kg dose volume of every kg body weight 0.6mg OCT, give rat through subcutaneous injection with the non-booklike particle precursor formulation of liquid.Before giving dosage, (give dosage preceding 1 day), give 10 minutes, 30 minutes, 1 hour, 3 hours, 6 hours, 24 hours and 48 hours blood sample collections (0.3mL) behind the dosage.

Like embodiment 16 said measurement OCT PCs.

The pharmacokinetics of OCT compares in the pharmacokinetics of the OCT for preparing in will the non-booklike particle in being scattered in saline and the saline solution.Result among Figure 16 shows that non-booklike particle dispersion provides the initial blood plasma level of significantly reduced OCT and (compares C with brinish situation _MaxReduce about 2.5 times), and discharge (or slow release) persistent period and be up at least 24 hours.

Embodiment 17-is used for other compositions of the OCT that is carried on non-booklike particle of injection (for example intravenous (i.v.), subcutaneous or intramuscular)

Described in embodiment 17, preparation contains the non-booklike particle dispersion of OCT (0.2mg OCT/mL) in saline.With the dispersion of gained muddy to milky with uniformly, and as through laser diffraction measurement, the about 100nm of particle mean size.Following table provides the composition of preparation.

The composition table that contains the non-booklike particle dispersion of OCT

Sample	Form (wt%)	Outward appearance
			2022OCT- A	SPC/GDO/P80/EtOH/OCT/ saline=2.71/4.06/1.19/0.89/0.02/91.13	All even white to light yellow dispersion
2022OCT- B	SPC/GDO/P80/DOPE-PEG (5000)/EtOH/OCT/ saline=2.47/4.06/1.19/0.24/0.89/0.02/91.13	All even white to light yellow dispersion

The syringe that utilization has No. 31 pins easily disperses non-booklike particle dispersion, or for example utilizes that pump sprayer easily sprays non-booklike particle dispersion.

Salmon calcitonin see calcimar (sCT) preparation in the non-booklike particle precursor of embodiment 18-liquid

Of embodiment 14, through the mixed SPC shown in the following table, GDO, P80, EtOH and sCT, preparation contains the non-booklike particle precursor of liquid (every g preparation contains 0.5mg sCT) of sCT.Through the sample of end-to-end rotation mixing gained, to obtain limpid and uniform liquid preparation.

The composition table that contains the non-booklike particle precursor of sCT

Sample ID	Form (wt%) SPC/GDO/P80/EtOH/sCT	Outward appearance
			2022sCT-A	28.78/43.18/17.99/10.00/0.05	Clarification, equal even light yellow liquids
2022sCT-B	30.58/45.88/13.49/10.00/0.05	Clarification, equal even light yellow liquids
			2022sCT-C	32.38/48.57/9.00/10.00/0.05	Clarification, equal even light yellow liquids
2022sCT-D	34.18/51.27/4.50/10.00/0.05	Clarification, equal even light yellow liquids

Embodiment 19-contains the lyophilization powder precursor of the non-booklike particle of OCT

Prepare the non-booklike particle precursor of liquid through mixing SPC (0.3046g), GDO (0.4570g), P80 (0.1344g), EtOH (0.100g) and OCT (0.004g), obtained limpid uniform liquid in 15 hours through end-to-end rotation subsequently.The Liquid precursor (0.50g) that will contain OCT is added in the 9.5g sterilized water, and on the jolting platform, mixes dispersions obtained 20 hours with 400rpm, to obtain the even non-booklike particle dispersion that every g preparation contains 0.2mg OCT.In non-booklike particle dispersion (9.0g), add the PVP solution of 9.0g 1wt%CMC aqueous solution and 18g 5wt%.The mixture of gained is added in the round-bottomed flask, and freezing on EtOH/ dry ice mixture, subsequently lyophilized overnight.The powder of gained be white to light yellow, have dry concordance and contain＜residual water of 2wt% and every g powder in OCT content be 1.3mg.Through vortex mixed powder easily is dispersed in the saline once more, to obtain the non-booklike particle dispersion of milky-white (muddiness).

The spray drying of the non-booklike particle of embodiment 20-

Through mixing the non-booklike particle dispersion of the SPC/GDO/P80 (31/54/15wt%) (5wt% amphiphilic substance) of preparation in advance among 6g such as the embodiment 1.1; With 12g1wt%CMC aqueous solution and 12g 5wt%PVP aqueous solution, obtain spray-dired non-booklike particle precursor.Adopt

mini spray exsiccator B-290 spray drying gained mixture; To obtain white to buff powder, said powder has the residual water of dry concordance and＜2wt%.Easily spray-dried powders is dispersed in the saline once more through vortex mixed, to obtain the non-booklike particle dispersion of milky-white (muddiness).

Non-booklike particle precursor of embodiment 21-liquid and the non-booklike particle dispersion that contains insulin

Through in vial, mixing SPC (0.918g), GDO (1.377g), P80 (0.574g) and EtOH (0.319g) and preparing the liquid fatty stock solution in 15 hours through end-to-end rotation subsequently.Insulin (10mg) is added in the sterilized water (0.190g), add 1.80g lipid stores solution in the insulin solution (every g preparation contains the 5mg insulin).Vortex gained mixture becomes evenly up to sample.

Lipid/insulin the mixture (1.85g) of above-mentioned preparation is added in the sterilized water (18.15g), and on vibration table with dispersion (the 0.46mg insulin/mL) 15 hours of 400rpm mixing gained.The dispersion of gained is muddy to milky and uniform.

Non-booklike particle precursor of embodiment 22-liquid and the non-booklike particle dispersion that contains GLP-1

Through in vial, mixing SPC (0.918g), GDO (1.377g), P80 (0.574g) and EtOH (0.319g) and, preparing the liquid fatty stock solution subsequently through end-to-end rotation 15 hours.GLP-1 (10mg) is added in the sterilized water (0.190g), add 1.80g lipid stores solution in the insulin solution (every g preparation contains the 5mg insulin).Vortex gained mixture becomes evenly up to sample.

Lipid/GLP-1 the mixture (1.85g) of above-mentioned preparation is added in the sterilized water (18.15g), and on vibration table with the dispersion (0.46mg GLP-1/mL) of 400rpm mixing gained 15 hours.The dispersion of gained is muddy to milky and uniform.

Claims (24)

1. particle composition, it comprises:

A) 10 to 40% at least a phosphatidylcholine component, said phosphatidylcholine component comprises at least a PC that is selected from synthetic dioleyl phosphatidyl choline, ovum PC, heart PC, brain PC, liver PC and Semen sojae atricolor PC,

B) 30 to 75% at least a DG component, at least a tocopherol or its mixture, wherein said DG component comprise the acyl chain with 14 to 18 carbon atoms DG and

C) 5 to 30% at least a nonionic stability amphiphilic substance; Said nonionic stability amphiphilic substance is lower than 8000 daltonian surfactants by molecular weight and forms; It is selected from stearic acid Polyethylene Glycol (PEG) ester; Distearyl acid PEG ester; Lauric acid PEG ester; Oleic acid PEG ester; The Oleum Ricini of polyethoxylated; PEG-DOPE; PEG-(4 hydroxy stearic acid ester); PEG-sorbitan-monolaurate; PEG-sorbitan-monopalmitate; PEG-sorbitan-monostearate; PEG-sorbitan-tristearate; PEG-sorbitan-monoleate; PEG-sorbitan-trioleate; PEG-sorbitan-single vaccenic acid ester and succinic acid d-alpha tocopherol base cetomacrogol 1000 ester

Wherein all umbers are with respect to the gross weight of a+b+c by weight, and wherein said compositions comprises the particle of at least a non-lamellar phase structure or form the particle of at least a non-lamellar phase structure when contacting with aqueous fluid.

2. the compositions of claim 1, wherein components b is the mixture of GDO or tocopherol and GDO.

3. the compositions of claim 1, wherein component a) and/or components b) derive from natural source.

4. the compositions of claim 1, wherein component a) has 50%C18:1 and/or C18:2 acyl group at least and a components b) be DG with 50%C18:1 at least and/or C18:2 acyl group.

5. each compositions, wherein amount of component b in the claim 1 to 4) comprise at least a nonionic stability amphiphilic substance that is selected from PEG-sorbitan-monolaurate and PEG-sorbitan-monoleate, PEG-(4 hydroxy stearic acid ester), succinic acid d-alpha tocopherol cetomacrogol 1000 ester and GREMAPHOR GS32.

6. the compositions of claim 1, it comprises activating agent in addition.

7. the compositions of claim 6, wherein said activating agent is be selected from the Sandostatin LAR Depot peptide relevant with other somatostatin, insulin, chlorhexidine digluconate, two hydrochloric acid chlorhexidines, diphosphate/ester, non-steroidal anti-inflammatory agent, corticosteroid, methotrexate, azathioprine, Ismipur and inhibitor of phospholipase enzymes at least a.

8. the compositions of claim 1, wherein said compositions comprises I ₂And/or L ₂The particle of phase structure and/or form I when contacting with aqueous fluid ₂And/or L ₂The particle of phase structure.

9. the compositions of claim 1 wherein comprises the particle of said compositions or the particle mean size of the particle that forms when contacting with aqueous fluid is 0.1 to 0.6 μ m.

10. the compositions of claim 1, wherein about phase behavior, granularity and particle size distribution, said composition stable at least 3 months.

11. the compositions of claim 1, it comprises in addition and is up to 20% at least a organic solvent and/or its water-soluble polymer, and wherein said organic solvent has 1 to 6 carbon atom, and said organic solvent is selected from alcohol, ketone, ester, ether and polymer thereof.

12. the compositions of claim 1; It comprises in addition and is up to 20% at least a organic solvent and/or its water-soluble polymer; Wherein said organic solvent has 1 to 6 carbon atom, and said organic solvent is selected from polyhydric alcohol, ketone, ester, ether and polymer thereof.

13. the compositions of claim 1, it is in following form:

A) dispersion,

B) preconcentrate in cosolvent,

C) dried powder or can tolerate the curing mixture of polymer with biology.

14. a pharmaceutical preparation, carrier or excipient that its at least a compositions and at least a biology that comprises in the claim 1 to 13 each can tolerate.

15. the pharmaceutical preparation of claim 14, it is in the following form that is selected from: the suspensoid in the liquid, powder formulation, tablet, capsule, aerosol, suppository, drop, cream and transdermal patch.

16. the pharmaceutical preparation of claim 14, it is in the form that is selected from coating capsule, coated tablet and spray.

17. the preparation of claim 14 or 15, it is suitable for parenteral and uses.

18. a method that forms non-booklike particle, it comprises the formation mixture, and said mixture comprises:

A) 10 to 40% at least a phosphatidylcholine component, said phosphatidylcholine component comprises at least a PC that is selected from synthetic dioleyl phosphatidyl choline, ovum PC, heart PC, brain PC, liver PC and Semen sojae atricolor PC,

B) 30 to 75% at least a DG component, at least a tocopherol or its mixture, wherein said DG component comprise the acyl chain with 14 to 18 carbon atoms DG and

C) 5 to 30% at least a nonionic stability amphiphilic substance; Said nonionic stability amphiphilic substance is lower than 8000 daltonian surfactants by molecular weight and forms; Said surfactant is selected from stearic acid Polyethylene Glycol (PEG) ester; Distearyl acid PEG ester; Lauric acid PEG ester; Oleic acid PEG ester; The Oleum Ricini of polyethoxylated; PEG-DOPE; PEG-(4 hydroxy stearic acid ester); PEG-sorbitan-monolaurate; PEG-sorbitan-monopalmitate; PEG-sorbitan-monostearate; PEG-sorbitan-tristearate; PEG-sorbitan-monoleate; PEG-sorbitan-trioleate; PEG-sorbitan-single vaccenic acid ester and succinic acid d-alpha tocopherol base cetomacrogol 1000 ester

Wherein all umbers are with respect to the gross weight of a+b+c by weight, and said mixture is dispersed in the aqueous fluid.

19. a test kit that is used to prepare the particle composition that is in the suspensoid form, said test kit comprise the description of each the described at least a compositions in the claim 1 to 13 of powder type and the optional said powder that in aqueous fluid, suspends.

20. each described compositions is used for treating the purposes of the medicine of human or animal's object in the claim 1 to 13 in preparation.

21. the purposes of claim 20, wherein said medicine is used to treat endoceliac inflammation and/or stimulation.

22. each purposes in the claim 20 to 21, wherein said medicine is used to treat inflammatory bowel.

23. each described compositions comprises the purposes in the preparation of at least a activating agent in preparation in the claim 1 to 13, wherein said preparation continues to discharge said activating agent 1 to 30 day.

24. each compositions is used for treating the purposes of the medicine of inflammation in the claim 1 to 13 in preparation.

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