CN101568328A - Microparticles based on an amphiphilic copolymer and on active ingredient(s) with modified release and pharmaceutical formulations containing same - Google Patents

Microparticles based on an amphiphilic copolymer and on active ingredient(s) with modified release and pharmaceutical formulations containing same Download PDF

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CN101568328A
CN101568328A CN 200780031893 CN200780031893A CN101568328A CN 101568328 A CN101568328 A CN 101568328A CN 200780031893 CN200780031893 CN 200780031893 CN 200780031893 A CN200780031893 A CN 200780031893A CN 101568328 A CN101568328 A CN 101568328A
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preferably
microparticles
po
ap
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CN 200780031893
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CN101568328B (en
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阿兰·孔斯坦西斯
陈幼平
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弗拉梅技术公司
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/14Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605Excipients; Inactive ingredients
    • A61K9/1629Organic macromolecular compounds
    • A61K9/1641Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/10Dispersions; Emulsions
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/0012Galenical forms characterised by the site of application
    • A61K9/0019Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/0012Galenical forms characterised by the site of application
    • A61K9/007Pulmonary tract; Aromatherapy
    • A61K9/0073Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy

Abstract

The present invention relates to novel microparticles of amphiphilic polyamino acids that transport active ingredient(s) - AI(s) -, in particular protein and peptide active ingredient(s), and also to novel modified-release pharmaceutical formulations containing said microparticles of AI. The objective of the invention is to develop novel microparticles loaded with AI, obtained by aggregation of nanoparticles of amphiphilic polyamino acids, and having improved properties, in particular in dry solid form, with regard to their capacity for dispersion and, as regards the reconstituted suspension, with regard to its stability and its ability to be readily handled and injected. In a first aspect, the invention relates to microparticles of amphiphilic polyamino acid (PO) containing at least one AI (associated noncovalently), spontaneously forming a colloidal suspension of nanoparticles in water, at pH 7.0, under isotonic conditions; wherein said microparticles are characterized a. in that they are obtained by atomization of a colloidal suspension or solution of PO comprising at least one AI, b. by a size of between 0.5 and 100 [mu]m, and c. in that they are dispersible in a colloidal suspension. The invention also relates to the method for preparing these microparticles, to a liquid formulation comprising a suspension of these PO/AI microparticles, to a method and a kit for reconstituting this formulation, and to a dry form of this formulation.

Description

基于两性分子共聚物和活性成分的改性释放的微颗粒以及含有它们的药物制剂技术领域 Based on amphiphilic copolymer and modified release of the active ingredient and microparticles BACKGROUND pharmaceutical formulations containing them

本发明涉及活性成分(AP)(尤其是蛋白质和肽活性成分的)新型的传送体,本发明还涉及含有所述AP传送体的改性释放的药物制剂。 The present invention relates to novel active ingredient (AP) carrying body (especially protein and peptide active ingredient), the present invention also relates to a pharmaceutical formulation comprising a modified release of the AP is transmitting member. 这些制剂有多种治疗(人用和兽用)用途。 These preparations have a variety of therapeutic (human and veterinary) use.

背景技术 Background technique

本说明书中所用的术语AP表示至少一种活性成分。 The term AP in this specification is represented by at least one active ingredient. 术语"改性释放"表示延长和域延迟和/或脉冲释放。 The term "modified release" means a field delay and extension and / or pulsatile release.

更具体地说,这种作为本发明目的的新型的AP传送体是由两性分子聚合物(例如疏水性基团改性的聚氨基酸)形成的微颗粒。 More specifically, the object of the present invention as novel microparticles AP transmitting member is formed from amphiphilic polymers (e.g., hydrophobic group modified poly amino acids). 这些微颗粒含有和所述聚合物结合的至少一种AP,并且能够以胶体悬浮液的形式或干燥形式提供。 These microparticles contain at least one AP and of the polymer-bound, and can be provided in dry form or in the form of a colloidal suspension.

在药用AP,尤其是治疗用肽/蛋白质的延长释放领域中,在很多情形下,目的是尽可能在患者体内重现接近健康受试者的观察值的血浆肽或蛋白浓度。 The AP in a pharmaceutical, in particular therapeutic peptides / proteins extended release art, in many cases, the purpose is to reproduce as close peptide or protein plasma concentration values ​​observed in healthy subjects in a patient.

这个目标与蛋白质在血浆里的存在期短相抵触,因此需要重复注射治疗蛋白质。 The target protein and the presence of short conflict in the plasma, it is necessary repeated injections of therapeutic proteins. 故治疗蛋白质的血浆浓度具有"锯齿"形,其特征是高浓度的峰和极低浓度的谷。 Therefore, the plasma concentration of therapeutic proteins having "sawtooth" shape, wherein the peaks and very low concentration of the high concentration of the valley. 峰浓度远大于健康受试者体内的基础浓度,由于治疗蛋白质(如白细胞介素IL2)的高毒性,其具有非常显著的有害效应。 Base peak concentration is much greater than the concentration of healthy volunteers, due to the high toxicity of therapeutic proteins (such as interleukin IL2), which has a very significant detrimental effects. 另外,谷浓度低于治疗效果所需的浓度,所以患者接受的治疗覆盖较少,而承受严重的长期副作用。 In addition, the trough concentrations below the desired therapeutic effect concentration, so fewer patients receiving treatment coverage, and bear serious long-term side effects.

因此,为在患者体内重现接近其治疗的理想值的治疗蛋白质血浆浓度,所讨论的药物制剂必须允许延长释放治疗蛋白质,以限制血浆浓度随着时间变化。 Therefore, in order to reproduce the plasma concentration of therapeutic protein close to its ideal value in a patient treated with the pharmaceutical formulation in question must allow the prolonged release of therapeutic proteins in order to limit the variations in plasma concentration over time.

而且,这种活性制剂优选应该满足以下为所属领域的技术人员所熟知的规 Furthermore, this active formulation should preferably meet the following of ordinary skill in the art in the field of regulatory

范: Fan:

1- 延长释放活性的和非变性的治疗蛋白质(例如人或合成蛋白质)以使血浆浓 1- prolonged release of the active and non-denatured therapeutic protein (e.g., human or synthetic proteins) to plasma concentration

度保持在治疗水平; Maintained at a therapeutic level;

2- 足够低的注射粘度以便于注射; 2- sufficiently low viscosity to facilitate injection of the injection;

3- 生物适应的和生物可降解的形式; 3- biocompatible and biodegradable form;

114- 无毒并且无免疫原性的形式; 114- toxic and non-immunogenic form;

5- 具有优良的局部耐受性的形式。 5- excellent topical tolerability.

为了实现这些目标,现有技术提出的最佳方法之一是开发治疗蛋白质的延长释放形式,其由加载治疗蛋白质的纳米颗粒的低粘度液体悬浮液构成。 To achieve these goals, one of the best prior art method proposed is the development of an extended release form of the protein therapeutic that low-viscosity liquid suspensions of nanoparticles loaded with therapeutic proteins configuration. 这些悬浮液有助于天然的治疗蛋白质的给药。 These suspensions facilitate administration of native therapeutic protein.

因此,所述治疗蛋白质和含有疏水性基团和亲水性基团的共聚氨基酸的纳米颗粒结合。 Thus, the therapeutic protein and amino acid copolymer comprising a hydrophobic group and a hydrophilic group bound nanoparticles.

专利US-B-5904936描述了一种两性分子聚氨基酸共聚物的平均尺寸0.01-0.5 pm的亚微米颗粒(NPV)和平均尺寸0.5-20 iim的微米颗粒(MPV),所述共聚物包括至少两种类型的氨基酸, 一种是中性和疏水性的,另一种是可电离的。 Patent US-B-5904936 describes the average size of a polyamino acid copolymer amphiphilic 0.01-0.5 pm submicron particles (NPV), and an average size of 0.5-20 iim microparticles (MPV), said copolymer comprising at least Two types of amino acids, one is neutral and hydrophobic, the other is ionizable. 在水性溶液中,蛋白质(如胰岛素)被自发地吸附到这些颗粒上。 In aqueous solution, proteins (such as insulin) are spontaneously adsorbed onto the particles. 这种聚氨基酸共聚物是例如聚(L-亮氨酸-b-(L-谷氨酸钠))的嵌段共聚物。 Such polyamino acid copolymer such as poly (L- leucine-b- (L- glutamate)) block copolymers. 该专利公开了通过向聚-Leu/Glu的胶体悬浮液加入单价阳离子盐(硫酸铵)或多价阳离子盐(Fe^、 Fe3+、 Zn2+、 Ca2+、 Al2+、 A产或Cu2+)、酸(HC1)或阳离子聚合物(聚赖氨酸)引发的NPV 向MPV的聚集。 This patent discloses the addition of a monovalent cation salt (ammonium sulfate) to poly -Leu / Glu colloidal suspension or a monovalent cation salt (Fe ^, Fe3 +, Zn2 +, Ca2 +, Al2 +, A production or Cu2 +), the acid (HC1 is) NPV aggregation induced or cationic polymers to MPV (polylysine).

专利申请WO-A-03/104303公开了两性分子的聚氨基酸,其含有天冬氨酸残基和/或谷氨酸残基,这些残基的至少一部分带有含有至少一个a-生育酚单元的接枝物(例如由合成或者天然来源的a-生育酚接枝的聚谷氨酸根或聚天冬氨酸根)。 Patent Application WO-A-03/104303 discloses amphiphilic polyamino acids comprising aspartic acid residues and / or glutamic acid residues, at least part of these residues having at least a unit a- tocopherol grafts (e.g. a- tocopherol grafted with synthetic or natural origin poly-glutamate or poly-aspartate). 在水中,这些"疏水改性的"均聚氨基酸自发地形成一种纳米颗粒胶体悬浮液,所述纳米颗粒在pH 7.4的水性悬浮液中易于和至少一种活性蛋白质(胰岛素) 相结合。 In water, these "hydrophobically modified" polyamino acids are spontaneously form a colloidal suspension of nanoparticles, said nanoparticles are easy and at least one active protein (insulin) in combination at pH 7.4 in aqueous suspension.

根据US-B-5904936和WO-A-2005/033181,由悬浮液"矢量化的"活性蛋白质(如胰岛素)的体内释放时间能够增加。 According to US-B-5904936 and WO-A-2005/033181, the suspension "vectorized" activity of a protein (e.g. insulin) in vivo release time can be increased.

在PCT申请WO-A-05/051416中公开的药物形式也部分实现了释放时间的增加。 Pharmaceutical forms disclosed in PCT application WO-A-05/051416 also partially achieved to increase the release time. 该专利申请公开了一种疏水改性的聚(L-谷氨酸钠)的纳米颗粒(0.001-0.5 pm) 的胶体悬浮液,其在一定浓度下注射,使得在皮下注射后,和内源性的白蛋白接触,在患者的注射部位形成凝胶体。 This patent application discloses a hydrophobically modified poly (L- sodium glutamate) nanoparticles (0.001-0.5 pm) colloidal suspension, injected at a certain concentration, such that after subcutaneous injection, and endogenous albumin contact formed gel at the injection site of the patient. 然后这种蛋白质缓慢释放,通常超过一周的时间。 This protein is then slowly released, usually more than a week. 然而,当给药的治疗蛋白质浓度相对高时,例如人生长激素的情形时,其释放时间只能局限在几天。 However, when relatively high concentrations of therapeutic proteins administered, for example, the case of human growth hormone, which is released only for a limited time in a few days.

通过提供一种用于AP延长释放的药物制剂有可能改进现有技术:•通过胃肠外途径(例如皮下)注射后,能够实现非变性并且高浓度(例 By providing a pharmaceutical formulation for the prolonged release of AP is possible to improve the prior art to: • the parenteral route (e.g. subcutaneous) injection, and possible to realize a high non-denaturing concentrations (Example

如几mg/ml)的AP (例如治疗用蛋白质、治疗用肽和小分子)体内释放时间的延长, As several mg / ml) of the AP (e.g. therapeutic proteins, peptides and small molecule therapeutics) prolonged in vivo release time,

•在物理化学和生理条件下均储存稳定。 • are stored chemical and physical stability under physiological conditions. 为了实现这个目的,可行的方法是按照WO 05/051416,通过加入和两性分子聚氨基酸的可电离基团(IG)极性相反的多价离子的方法将制剂的纳米颗粒聚集在一起以提供一种微颗粒,所述离子以确定的比例存在。 For this purpose, a method is possible in accordance with WO 05/051416, and by the addition of amphiphilic polyamino acids ionizable group (IG) method of multivalent ions of the polarity of the formulation nanoparticles aggregated together to provide a reverse microparticles species, to determine the ratio of the ion is present. 这导致特定微颗粒群体的选择,结合有AP的微颗粒能够使所述AP(例如蛋白质或肽)释放的时间得到显著延长。 This results in the selection of a specific population of microparticles, AP bound microparticles enables the AP (e.g. protein or peptide) release time is significantly extended. 一些多价离子,例如Mg2+、 Ca2+、 Zn2+、 Fe2+、 Ci^+或它们的混合物和/或A产、F^+或它们的混合物使得这种用于AP延长释放的液体药物制剂具有极好的耐受性。 Some multivalent ions such as Mg2 +, Ca2 +, Zn2 +, Fe2 +, Ci ^ + or mixtures thereof and / or a A yield, F ^ + or mixtures thereof such that such liquid pharmaceutical formulation AP extended release for excellent tolerance. 例如,这种制剂可以含有基于合成来源的a-生育酚接枝的粒径为0.5-100 pm的聚谷氨酸微米尺寸的颗粒的低粘度的水性胶体悬浮液,其含有多价离子Mg2+、 Ca2+、 Zn2+、 Fe2+、 Cu2+、 Al3+或Fe3、其乘积r,如下式所定义,为0.3-10, For example, such preparations may contain synthetic origin, based on a- tocopherol particle diameter of grafted low-viscosity aqueous colloidal suspension of particles of micron size polyglutamate 0.5-100 pm, which contains multivalent ions Mg2 +, Ca2 +, Zn2 +, Fe2 +, Cu2 +, Al3 + or with Fe3, which is the product of r, as defined in formula, 0.3 to 10,

g巾 g towel

,n是所述多价离子的化合价, , N being the valence of polyvalent ions,

■ [MI]是多价离子的摩尔浓度, ■ [MI] is the molar concentration of multivalent ions,

■ [IG]是可电离基团IG的摩尔浓度。 ■ [IG] is the molar concentration of ionizable groups IG. 这些选定的微米尺寸的颗粒来自大量两性分子共聚物纳米颗粒的聚集。 The micron-sized particles selected from a large accumulation of amphiphilic copolymer nanoparticles. 加载AP(例如hGH)的微颗粒的悬浮液因此能够通过和多价离子Mg2、 Ca2+、 Load suspension AP (e.g. hGH) The microparticles can be adopted and multivalent ions Mg2, Ca2 +,

Zn2+、 Fe2+、 Cu2+、 A产或Fe"的絮凝作用以及随后的陈化和洗涤生产。这种悬浮液可以随后冻干或雾化并且然后可以和水重构以产生一种能够现成用于注射的制剂。 Zn2 +, Fe2 +, Cu2 +, A production or Fe "flocculation and subsequent production of washing and aging. This suspension may then be lyophilized or atomized water and then reconstituted to produce a ready-made for injection can be preparations.

很明确这些干燥粉末形式的微颗粒在储藏方面具有优势。 These very clear dry powder form of micro-particles have advantages in terms of storage. 具体地说,它们应当能够增加这些微颗粒的物理稳定性和AP的稳定性。 In particular, they should be able to increase the stability and the physical stability of the AP microparticles.

然而,在通过胃肠外途径将这些微颗粒粉末用于治疗应用时,难度在于当使用时能够容易地分散(或悬浮)这些粉末以获得重构的液体微颗粒形式,更具体地说,是一种悬浮液,它在室温下是稳定的,例如至少稳定几分钟,事实上平均至少稳定几十分钟。 However, when by parenteral route these micro-particles for therapeutic applications, the difficulty is that when used can be easily dispersed (or suspension) of these powders to obtain a liquid form reconstituted microparticles, more particularly, one kind of suspension, it is stable at room temperature and stable for at least several minutes, for example, an average of actually stable for at least several tens of minutes. 这种稳定性是人们想要的,尤其是为了能够易于操作这种重构的悬浮液。 This stability is that people want to be able to particularly easily reconstituted operation of such a suspension. 这种悬浮液能够易于通过连接在注射器或注射笔(胰岛素笔类型)上的空心针头而容易地注射也很重要。 This suspension can be easily connected to a syringe or by injection pen (insulin pen type) of the hollow needle is easily injection is also important. 最后,还应当考虑到一个事实,即分散、操作和注射这种重构的悬浮液将由患者或医务人员进行。 Finally, it should also take into account the fact that the dispersion, and the operation of such an injection suspensions reconstituted by the patient or medical personnel.

发明内容 SUMMARY

在这种情形下,本发明的一个主要的目的是提供一种新型的微颗粒,其通 In this case, a main object of the present invention is to provide a novel microparticle that pass

过可生物降解的和水溶性的两性分子的聚合物的纳米颗粒(例如根据WO-A-05/051416中公开的那些制剂的类型)的聚集获得,这些微颗粒加载有AP,并且能够显示出改进的性质,尤其是干燥的固体形式,特别是关于它们被分散的能力("可分散性"),以及对于重构的悬浮液,其稳定性以及其易于操作和注射的能力。 Biodegradable polymers too water soluble and amphiphilic nanoparticles (e.g., those formulations in accordance with the type of WO-A-05/051416 disclosed) to obtain aggregated, microparticles loaded with the AP, and can exhibit improved properties, especially the dry solid form, in particular for a suspension of reconstituted, its stability, and is easy to handle and injection capabilities on their ability to be dispersed ( "dispersibility"), as well.

本发明的另一个目的是提供一种微颗粒,其通过可生物降解的和水溶性的两性分子的聚合物的纳米颗粒的聚集获得,所述微颗粒加载有AP,并且具有良好的分散性质,无论是在含水相中或者是在有机相中,同时保持所述微颗粒的完整性。 Another object of the present invention is to provide a microparticle, nanoparticles aggregated polymer degradation by water-soluble and biodegradable amphiphilic molecules obtained, the microparticles loaded with the AP, and has excellent dispersion, whether the aqueous phase or in organic phase, while maintaining the integrity of the microparticles.

本发明的另一个目的是提供一种微颗粒,其通过可生物降解的和水溶性的两性分子的聚合物的纳米颗粒的聚集获得,所述微颗粒加载有AP,并且其固体和干燥形式具有优良的稳定性。 Another object of the present invention is to provide a microparticle, nanoparticles aggregated polymer degradation by water-soluble and biodegradable amphiphilic molecules obtained, the microparticles loaded with the AP, and a solid and dry form having excellent stability.

本发明的另一个目的是提供一种制备固体和干燥形式微颗粒的方法,而且这些微颗粒: Another object of the present invention is to provide a solid preparation and drying methods in the form of microparticles, and the microparticles:

— 通过可生物降解的和水溶性的两性分子的聚合物的纳米颗粒(例如根据WO-A-05/051416中公开的那些制剂的类型) 的聚集获得, - (e.g., those formulations in accordance with the type of WO-A-05/051416 disclosed) obtained through the aggregation of nanoparticles of a biodegradable polymer and water-soluble amphiphilic,

~> 加载有AP, ~> Loaded with AP,

— 并且如上述的目的所定义。 - as defined above and the object. 本发明的另一个目的是提供一种制备固体和干燥形式微颗粒的方法,其类 Another object of the present invention is to provide a solid preparation in the form of dried microparticles and methods of its class

型为前面的目的中所定义,而且这种方法简单、经济以及是工业性质的。 Type as defined in the preceding object, and this method is simple, economical and industrial properties.

本发明的另一个目的是提供一种药物制剂,其含有新型的微颗粒,所述微颗粒通过可生物降解的和水溶性的两性分子的聚合物的纳米颗粒(例如根据WO-A-05/051416 中公开的那些制剂的类型)的聚集获得,而且这些微颗粒:4 加载有AP, — 并且如上述的目的所定义。 Another object of the present invention is to provide a pharmaceutical formulation containing the novel microparticles, nanoparticles of the polymer microparticles by a biodegradable and amphiphilic water-soluble (e.g. according to WO-A-05 / those of the type disclosed in the formulation of 051,416) to obtain aggregates, and these microparticles: 4 loaded with AP, - and the object as defined above. 本发明的另一个目的是提供一种药物制剂,其用于AP的延长释放,该制剂能够克服现有技术的缺陷,并且尤其能够在通过胃肠外途径(例如皮下)注射后, Another object of the present invention is to provide a pharmaceutical formulation for the prolonged release of the AP, the formulation is capable of overcoming the drawbacks of the prior art, and in particular after the injection can be by parenteral route (e.g. subcutaneous),

使非变性的AP(例如蛋白质、治疗用肽或小分子)的体内释放时间得到延长。 Non denatured AP (e.g. proteins, therapeutic peptides or small molecules) to give prolonged in vivo release time.

本发明的另一个目的是提供一种药物制剂,其能够在通过胃肠外途径(例如皮下)注射后,使高浓度(例如几mg/ml)的治疗用蛋白质或肽的体内释放时间得到延长。 Another object of the present invention is to provide a pharmaceutical formulation, which can be by parenteral route after (e.g. subcutaneous) injection, high-concentration (e.g., several mg / ml) with time in vivo release of therapeutic protein or peptide to be extended .

本发明的另一个目的是提供一种药物制剂,用于延长AP的体内释放时间, 其在物理化学和生理条件下均储存稳定。 Another object of the present invention is to provide a pharmaceutical formulation for the prolonged release of AP in vivo time, which are stored in the physical and chemical stability and physiological conditions.

本发明的另一个目的是提供一种药物制剂,用于延长AP的体内释放时间, 其显示出以下性质中的至少一种:生物相容性、生物可降解性、无毒性和良好的局部耐受性。 Another object of the present invention is to provide a pharmaceutical formulation for prolonging the in vivo release time of the AP, which exhibits at least one of the following properties: biocompatibility, biodegradability, non-toxicity and good resistance to localized sexual.

本发明的另一个目的是提供一种药物制剂,用于延长AP的体内缓慢释放时间,所述制剂含有和至少一种AP自结合的两性分子聚合物PO的微颗粒(PO/AP 微颗粒),所述聚合物PO是带有疏水性基团(HG)和亲水性基团[优选至少部分电离的可电离基团(IG)]的水溶性可生物降解的聚合物,其在水中自发地形成胶体纳米微粒的悬浮液,这种聚合物PO是,例如一种聚氨基酸,其主链由天冬氨酸残基或谷氨酸残基形成,至少部分这些单元通过在链中和/或链端接枝至少一个疏水性基团HG而改性。 Another object of the present invention is to provide a pharmaceutical formulation for the slow prolonged release of AP in vivo time, the formulation containing the amphiphilic polymer microparticles of PO and at least one self-association of the AP (PO / AP microparticles) , PO is a polymer having a hydrophobic group (HG) and the hydrophilic group [preferably at least partially ionized ionizable group (the IG)] of water-soluble biodegradable polymers, which spontaneously in water forming a colloidal suspension of nanoparticles, the polymer PO is a polyamino acid such as one whose main chain is formed by an aspartic acid residue or glutamic acid residues, at least partially by these units in the chain and / branched chain or terminating the at least one hydrophobic group HG be modified.

本发明的另一个目的是提供一种药盒,用于重构前面所列目的所定义的制剂,这种药盒例如是使用简单的因此能够由病人或医务人员容易地使用。 Another object of the present invention is to provide a kit for reconstructing the front formulations listed object defined, for example, such a kit is simple to use can be easily used by the patient or medical personnel.

本发明的另一个目的是提供一种重构如前面所列目的所定义的制剂的方法,这种方法是,例如,操作简单的,尤其是对于病人或医务人员。 Another object of the present invention is to provide a method for preparation purposes as set out hereinbefore defined reconstructed, this method is, for example, simple operation, especially for the patient or medical personnel.

本发明的另一个目的是提供一种固体药物制剂,用于AP的延长释放,尤其是一种用于肺部吸入和给药的干燥粉末形式: Another object of the present invention is to provide a solid pharmaceutical formulation for the prolonged release of AP, in particular a dry powder inhalation and pulmonary administration for:

•其基于如前面所列目的定义的和至少一种AP结合的PO微颗粒; •或者由如前面所列目的定义的制剂获得。 • PO based microparticles as previously listed and defined at least one object AP binding; • or object as defined by the formulations listed earlier obtained.

为了实现这些目的,其中,在长期的勤奋研究之后,发明人的贡献在于以完全意外和预想不到的方式,发现了雾化基于两性分子PO(例如基于共聚氨基酸)和基于AP的制剂将获得非常稳定的干燥PO/AP微颗粒,其能够实现在液体介 To achieve these objects, which, after a diligent study of long-term, the contribution of the inventors that the unexpected and completely unexpected manner, based on the atomization of amphiphilic molecules found PO (e.g. amino acid-based copolymer) and an AP-based formulation is obtained stable dry PO / AP microparticles, which can be implemented in liquid medium

质中重构0.5-100微米大小的微颗粒的水性悬浮液。 Mass reconstituted microparticle size of 0.5 to 100 microns an aqueous suspension.

而且,发明人还对于开发从这些微颗粒重构悬浮液的方法作出了贡献,所述方法在水性液体相和有机液体相中都能够优化它们的分散。 Further, the inventors have also contributed to the development of these methods from a suspension of microparticles is reconstituted, the method can optimize their phase dispersed in an aqueous liquid and organic liquid.

这是因为优质稳定易于获得的分散体是把这些重构自干燥PO/AP微颗粒的悬浮液用作可注射制剂的前提。 This is because the quality and stability of the dispersion is easily obtained from these reconstituted dried premise PO / AP microparticles suspension was used as an injectable formulation.

雾化是人们所知的从由颗粒作为成分材料的溶液或悬浮液生产干燥颗粒的方法。 It is known to spray dried particles produced by the process from the material component particles as a solution or suspension. 雾化(或喷雾干燥)是这种溶液或悬浮液的雾状小液滴在热空气流或热的惰性气体流中非常快的蒸发。 Atomizing (or spray dried) which is a mist of small droplets of a solution or a suspension in an inert gas stream of hot air or hot very quickly evaporated.

在制药领域,雾化可能由于其带入了热应力而成为问题,所述热应力证明对一些热敏感的AP完全不适用,例如基于蛋白质或其它肽化合物的AP。 In the pharmaceutical field, which may be due to atomization into a problem of thermal stress, the thermal stress demonstrated some heat sensitive AP completely NA, for example based on a protein or peptide compound other AP. 因此,依靠雾化以获得基于赋形剂和AP的干燥颗粒不是显而易见的,除非通过选择能够避免肽AP变性(实际上是使其最小化)的赋形剂。 Thus, relying on the atomization in order to obtain dry particles of AP and excipients not obvious, unless the peptides AP can be avoided by selecting degeneration (actually minimizes) excipient.

因此,专利申请US-A-2005/0158392公开了通过雾化加载了肽AP的固体亲水性微颗粒的制备方法。 Thus, patent application US-A-2005/0158392 discloses a method for preparing a peptide loading atomizing AP solid hydrophilic microparticles. 这种雾化在于雾化一种水性溶液,其含有透明质酸、 AP和亲脂性表面活性齐tj(例如卵磷脂),事实上甚至另一种Tween® 80类型的表面活性剂,或者,在第一步,雾化一种水性溶液,其含有透明质酸、AP和可选的一种Tween® 80类型的表面活性剂以获得一种初级颗粒,然后在第二步,雾化一种亲脂性表面活性剂(例如卵磷脂)的醇性溶液,其中分散了所述的初级颗粒。 Such atomizing spray comprising an aqueous solution containing hyaluronic acid, AP, and a lipophilic surfactant Qi TJ (e.g., lecithin), indeed even of another type of Tween® 80 surfactant, or, in the step, atomizing an aqueous solution containing hyaluronic acid, AP, and optionally one type of Tween® 80 surfactant to obtain a primary particle, and then in a second step, atomization one lipophilic alcoholic solution of a surfactant (e.g., lecithin), wherein said primary particles are dispersed. 该美国申请的重点在于透明质酸和亲脂性表面活性剂(例如卵磷脂)的保护性联合,后者的目的在于形成一层用于基于透明质酸和基于AP的微颗粒的覆盖膜。 The key is that the US application of hyaluronic acid and a lipophilic surfactant (e.g., lecithin) protective joint, which aims to form a hyaluronic acid-based film and a cover layer for the AP-based microparticles. 这份文件既未提及甚至也没有暗示两性分子PO的使用,以及两性分子的共聚氨基酸作为AP的赋形剂或者传送体和AP形成微颗粒。 This document neither mentioned nor suggested even amphiphilic PO use of amino acids and copolymers of amphiphilic molecules is formed as microparticles or excipient transfer member AP and AP.

在Maa等人J. Pharm. Sci., 87(2), 152-159, 1988的论文中描述了其它的雾化例子。 ., 87 (2), 152-159, 1988 describes other articles atomization examples Maa et al. J. Pharm. Sci. 根据该论文,所述AP(人生长激素)在雾化以产生微颗粒之前在生物可降解的表面活性剂存在下和锌复合。 According to this paper, the AP (human growth hormone) and zinc complexes in the presence of a surfactant biodegradable before atomization to produce fine particles.

因此,很明显蛋白质雾化的操作很困难,它要求使用含有多种赋形剂的复合制剂,以在过程中保护所述AP并且保证其储存的稳定性,这一点对于一些敏感的分子,如人生长激素(hGH)尤其是非常重要的。 Thus, it is clear that proteins atomizing operation is difficult, it requires complex formulation containing more excipients, in the process in order to protect and ensure the stability of the AP storage thereof, which is sensitive for some molecules, such as human growth hormone (hGH) in particular is very important.

而且,由于现有技术没有教导关于设计用于AP延长释放的干燥微颗粒在液体中分散和稳定的难题(尤其是关于两性分子PO的微颗粒),发明人的贡献是巨大的。 Further, since the prior art does not teach about the problems designed for extended release AP dried microparticles dispersed and stabilized in the liquid (especially amphiphilic molecules on microparticles PO), the inventor has contributed enormously.

本发明的第一方面涉及含有至少一种活性成分(AP)的聚合物(PO)的微颗粒,所述聚合物PO The first aspect of the invention relates to microparticles comprising at least one active principle (AP) polymer (PO), the polymer PO

,是带有疏水性基团(HG)和亲水性基团的水溶性生物可降解的两性分 , Having a hydrophobic group (HG) and a water-soluble hydrophilic group of bio-degradable fraction amphoteric

子(共)聚合物, Promoter (co) polymer,

,在pH7.0,等渗条件下,在水中自发地形成纳米颗粒的胶体悬浮 , At pH 7.0, isotonic condition, in water to form spontaneously the nanoparticles colloidal suspension

液, liquid,

•并且和所述AP非共价地结合; 其中微颗粒 • and the AP and non-covalently bound; wherein microparticles

a. 通过雾化含有至少一种AP的PO的溶液或胶体悬浮液获得, a. atomizing obtained by containing at least one AP-PO solution or colloidal suspension,

b. 以T试验测得的尺寸为0.5-100微米,优选l-70微米,更优选2-40微 b. T test as measured in a size of 0.5 to 100 microns, preferably l-70 [mu] m, more preferably 2-40 micro

米, Meter,

c. 并且在"可分散性"试验DP 1中在胶体悬浮液中是可分散的。 C. and in "dispersibility" DP 1 in Test colloidal suspension is dispersible. 本发明第二方面涉及一种制备PO微颗粒的方法,其与至少一种活性成分 The second aspect of the invention relates to a method for preparing microparticles PO, with at least one active ingredient

(AP)相结合,这些PO/AP微颗粒尤其是根据本发明第一方面所定义的那些, i该聚合物PO (AP) in combination, the PO / AP microparticles as defined according to a first particular aspect of the present invention are those, the polymer PO I

• 是带有疏水性基团(HG)和亲水性基团[优选至少部分电离的可电离基团(IG)]的水溶性可生物降解的两性分子(共)聚合物, • with a water-soluble hydrophobic group (HG) and the hydrophilic group [preferably at least partially ionized ionizable group (the IG)] biodegradable amphiphilic molecules (co) polymer,

• 在pH7.0,等渗条件下,在水中自发地形成纳米颗粒的胶体悬浮液, • at pH 7.0, isotonic conditions, spontaneously in water forming a colloidal suspension of nanoparticles,

• 并且和所述AP非共价地结合; • and the AP and non-covalently bound;

ii所述微颗粒以T试验测得的尺寸为0.5-100微米,优选l-70微米,更优选2-40微米,其主要包括雾化含有AP的PO的溶液或胶体悬浮液。 ii the microparticles to test T dimension measured from 0.5 to 100 microns, preferably l-70 [mu] m, and more preferably 2-40 microns, which includes atomizing a solution containing an AP PO or colloidal suspension. 根据一个有趣的实施方式,所述通过雾化获得的微颗粒再分散于基本上是水性的液体介质中(优选含有多价离子作为分散方法),然后冻干所获得的分散体。 According to an interesting embodiment, the microparticles obtained is atomized by re-dispersed in a substantially aqueous liquid medium (preferably containing a polyvalent ion as a dispersion method), and then the obtained dispersion was freeze-dried. 本发明第三方面涉及一种液体药物制剂,用于AP的延长释放,其含有一种"低"粘度的胶体悬浮液,所述胶体悬浮液基于含有至少一种AP的PO微颗粒,这些微颗粒是根据本发明第一方面所定义的那些或者是根据本发明第二方面所定义的方法获得的那些。 The third aspect of the present invention relates to a liquid pharmaceutical formulation for the prolonged release of the AP, which contains a "low" viscosity colloidal suspension, a colloidal suspension based on PO microparticles comprising at least one AP, the microstructures or those particles are those defined in the method according to the second aspect of the present invention as defined in the first aspect of the present invention is obtained. 本发明第四方面涉及一种重构药盒,尤其用于重构根据本发明第三方面定义的制剂,其包括- A fourth aspect of the present invention relates to a reconstruction kit, in particular for reconstructing a third aspect of the definition of the formulation according to the invention, which comprises -

•含有至少一种AP的PO微颗粒,这些微颗粒是根据本发明第一方面所定义的那些或者是根据本发明第二方面所定义的方法获得的那些; ,和重构液体,其选自以下组: • PO containing at least one kind of microparticles of AP, microparticles or those that are obtained according to the method of the second aspect of the present invention, as defined as defined according to a first aspect of the present invention; and reconstitution liquid, which is selected from the following groups:

4 基本上水性的液体; 4 substantially aqueous liquid;

4 基本上有机并且与水互溶的液体; 4 and the substantially water-miscible organic liquid;

— 以及基本上有机并且与水不互溶的液体。 - and and a substantially water-immiscible organic liquids. 本发明第五方面涉及一种重构方法,尤其用于重构根据本发明第三方面定 The fifth aspect of the present invention relates to a reconstruction method for reconstructing a particular set according to a third aspect of the present invention

义的制剂,其主要包括: Meaning preparations which include:

•混合 •mixing

3含有至少一种AP的PO微颗粒,这些微颗粒是根据本发明第一 3 PO microparticles comprising at least one AP, the microparticles of the present invention according to a first

方面所定义的那些或者是根据本发明第二方面所定义的方法获得的那些. Or those those obtained according to the method of the second aspect of the present invention as defined in aspect defined above.

3和一种重构液体,其选自以下组: 3 and one reconstitution liquid, which is selected from the group:

— 基本上水性的液体; - a substantially aqueous liquid;

— 基本上有机并且与水互溶的液体; - substantially miscible with water and organic liquid;

— 以及基本上有机并且与水不互溶的液体; ,并且搅拌这种混合物。 - and and a substantially water-immiscible organic liquid;, and this mixture was stirred.

本发明第六方面涉及一种固体药物制剂,用于AP的延长释放,其含有用于吸入和肺部给药的干燥的粉末形式: Sixth aspect of the invention relates to a solid pharmaceutical formulation for the prolonged release of AP that forms containing a dry powder for inhalation and pulmonary administration:

參基于含有至少一种AP的PO微颗粒,这些微颗粒是根据本发明第一方面所定义的那些或者是根据本发明第二方面所定义的方法获得的那些; ,或者根据本发明第三方面所定义的制剂获得。 Reference PO based microparticles comprising at least one AP, the microparticles according to the first aspect of the present invention are those defined or those obtained according to the method of the second aspect of the present invention, as defined above; or, according to a third aspect of the present invention, formulation as defined is obtained. 优点: advantage:

不愿被理论束缚,也可以认为能够聚集起来以提供疏水性区域的所述聚合物PO的疏水性基团在所述AP改性释放的过程中起了重要的作用,如同在后者的稳定中一样。 Without wishing to be bound by theory, believed to be aggregated together to provide hydrophobic regions in the polymer PO is a hydrophobic group plays an important role in the modified release of the AP, the latter as stable in the same.

也可以假设所述两性分子聚合物PO的疏水区域和所述AP(尤其是蛋白质类 May be assumed that the amphiphilic polymer and the hydrophobic regions PO AP (especially proteins

型的)之间发生了物理(非共价滩互作用。所述蛋白质对于所述聚合物的强亲和 It occurred between type) Physical (non-covalent interaction beach. Strong affinity of the protein for the polymer

18力导致皮下给药后其释放延长。 18 force causes prolonged release after subcutaneous administration. 这种释放机理尤其和聚乳酸、聚乳酸-乙二醇酸的微颗粒中观察到的不同,并且可以可选地与聚乳酸、聚乳酸-乙二醇酸的微颗粒中观察到的结合起来,或者也可以和透明质酸钠的那些结合起来,其中所述AP的释放主要与所述活性成分的扩散和所述颗粒的分解/蚀耗有关。 Such release mechanism and in particular, polylactic acid, polylactic acid - glycolic acid microparticles observed different, and may optionally polylactic acid, polylactic acid - glycolic acid combination of microparticles observed or may be those of sodium hyaluronate and consumption relevant combination, wherein the release of the AP and the primary particles and the diffusion of the active ingredient decomposition / corrosion.

所述蛋白质对于所述聚合物PO的亲和力使得能够限制蛋白质的聚集现象以及其它在雾化过程中可能出现的破坏性情况的发生,而不必加入其它稳定剂(例如糖、表面活性剂等)。 For the protein affinity such that the polymer PO is possible to limit the occurrence of protein aggregation, and other destructive situation may occur in the atomization process, without addition of other stabilizers (e.g., sugars, surfactants, etc.). PO的这种保护效果还使得能够容易获得储存稳定的液体制剂。 This protective effect PO also enables easy access to a storage stable liquid formulation. 最后,当PO主要是无定形性质时(尤其当PO是聚氨基酸时),当它们以干燥形式储存时,这给所述微颗粒带来优异的物理稳定性。 Finally, when the PO predominantly amorphous properties (particularly when PO is a polyamino acid), when they are stored in dry form, which results in superior physical stability to the microparticles.

因此,很明显这些两性分子聚氨基酸PO的存在带来了控制所述AP改性释 Thus, it is clear that the control of the AP brings these modified release is present in the amphiphilic polyamino acids PO

放以及关于聚集或者可能的化学分解的稳定化的新方法。 And put new methods of stabilizing chemical decomposition on the aggregated or possible.

而且,所述聚合物PO的两性分子性质使得能够在雾化过程中完全使用含水 Furthermore, the amphiphilic properties of the polymer PO enables full use of the atomization process in an aqueous

相或者完全使用有机挥发相或使用含水和有机相的挥发性混合物,这给方法的实施带来了极大的灵活性。 Volatilization of the organic phase or completely using a volatile or a mixture of an aqueous phase and an organic phase, to which the method of the embodiment brings great flexibility.

'而且,由于其化学性质,这些由所述(共)聚氨基酸型PO形成的微颗粒是生 'Furthermore, due to its chemical nature, these micro-particles formed by the (co) PO is a polyamino acid type Health

物可降解的和生物相容性的,并且总体来说具有良好的局部耐受性。 Biodegradable and biocompatible, and overall good local tolerance.

关于制备根据本发明的所述固体和干燥微颗粒的方法,即雾化,其在本质 For the preparation according to the present invention, the solid microparticles and drying methods, i.e. atomized, which in essence

上也容易转变成工业化规模。 Also easily transformed into the industrial scale.

最后,本发明的微颗粒以及含有它们的制剂是无毒的并且局部耐受良好。 Finally, the microparticles of the present invention and formulations containing them are non-toxic and well tolerated locally.

定义: definition:

在本申请的全文中,连词"或"应当理解为包括"这个或另一个或两者"的意 In the present application text, the conjunction "or" should be understood to include the meaning "one or the other or both," the

思。 think. 因此,例如可以含有至少一个杂原子(O、 N或S)或至少一个不饱和键的直链烷基可以具有两个杂原子,N和S,和一个不饱和键。 Thus, for example, can contain at least one heteroatom (O, N or S) at least one unsaturated bond or a straight chain alkyl group may have two hetero atoms, N and S, and one unsaturated bond.

在本申请的全文中,术语"亚微米-等级的颗粒"或"纳米颗粒"和根据本发明的微颗粒不同,指颗粒尺寸(以下面描述的T试验测定)为,例如大于或等于l nm 并且小于500nm,优选5-250nm。 In the present application text, the term "sub-micron - grade particles" or "nanoparticle" and in accordance with the microparticles of the present invention is different, refers to a particle size (T test measures the manner described below), for example greater than or equal to l nm and less than 500nm, preferably 5-250nm.

在本申请的全文中,根据本发明,术语"聚氨基酸"同时覆盖天然的聚氨基酸与合成的聚氨基酸,以及含有2-20个氨基酸残基的低聚氨基酸,和含有大于20个氨基酸残基的聚氨基酸。 In the present application in full, according to the present invention, the term "polyamino acid" covers both natural and synthetic polyamino acids polyamino acid, and oligomeric acid contains 2-20 amino acid residues, and containing more than 20 amino acid residues poly amino acids.

聚氨基酸主链优选的氨基酸残基是具有L构型的那些,键类型优选a型的,键存在于氨基酸的a氨基和另一个a-氨基酸的l位的羧基之间。 Preferred polyamino acid backbone having amino acid residues are those of the L configuration, preferably a type of the key type, bond between a carboxyl group present in the amino acid at position l a- a-amino group and another amino acid.

根据本发明,术语"蛋白质"指例如蛋白质和肽,无论其为寡肽或多肽。 According to the present invention, the term "protein" refers to proteins and peptides, e.g., whether it is an oligopeptide or polypeptide. 这种蛋白质或这种肽可以是改性或未改性的,例如,通过接枝一个或多个聚乙二醇基团改性。 Such proteins or peptides that may be modified or unmodified, e.g., by grafting one or more modified polyethylene glycol group.

在本申请的全文中,根据本发明,术语"结合"(名词或动词)用于描述一种或 In the present application in full, according to the present invention, the term "binding" (noun or verb) is used to describe one or

多种AP和聚合物PO之间的关系,尤其是指所述活性成分或多个成分通过非共价键和一种或多种聚合物PO(s)结合,例如通过静电或疏水相互作用或氢键或立体相互位阻。 The relationship between the AP and the plurality of polymer PO, particularly to the active ingredient or a plurality of components joined by non-covalent bonds and one or more polymers PO (s), for example by electrostatic or hydrophobic interactions or hydrogen bonding or steric another perspective.

本发明的第一方面:微颗粒特.征M:… The first aspect of the present invention: microparticle characteristics M:. ...

在雾化之前,AP(例如一种蛋白质或其它肽化合物)和一种两性分子聚合物PO(例如一种两性分子(共)聚氨基酸)的结合事实上显示出许多同时关于方法本身和通过雾化生产的微颗粒的性质方面的优点。 Prior to atomizing, binding AP (e.g. a protein or another peptide compound) and one amphiphilic polymers PO (for example, an amphoteric molecule (co) poly-amino acids) showed virtually simultaneously on many mist process itself and by nature of the advantage of production of microparticles.

物理雾化处理可以使所得的干燥固体微颗粒具有独特的结构,其带来了大量优良的性质。 Physical treatment may atomizing the resulting solid was dried microparticles have a unique structure, which brings a lot of excellent properties. 这种结构通过这种雾化的生产方法以及这些优良的性质所附带的功能正确地进行表征。 This configuration properly be characterized by the production method of such atomization and these excellent properties of the attendant functions.

授.征.0?〕;.... .] .0 levy granted;? ....

该特征由下面描述的T试验客观地定义。 T Test This feature is defined by the following described objectively.

通过激光衍射测定微颗粒尺寸的T试验: Micro determined by laser diffraction particle size T test:

a-TO试验,其中微颗粒是干燥形式的1设备和操作条件 a-TO test, wherein the microparticle is a drying apparatus and operating conditions in the form of

激光颗粒检测仪 Malvern Mastersizer 2000 Laser particle detector Malvern Mastersizer 2000

单位 Hydro 2000SM液体通路 Hydro 2000SM liquid path unit

分散载体流体体积 150 ml Dispersing a carrier fluid volume 150 ml

波长(蓝和红) 466和632 nm Wavelength (blue and red) and 632 nm 466

搅拌速度 2400 rev/min Stirring speed of 2400 rev / min

分析范围 0.02 ,2000 0.02 Analysis, 2000

光学模型(Mie理论) Optical model (the Mie theory)

所使用的折射率值: n流体=1.39 + i0 Refractive index value used: n = 1.39 + i0 fluid

分散流体(水) 11聚苯乙烯胶乳=1.59 + iO Dispersing fluid (water) 11 = 1.59 + iO polystyrene latex

聚苯乙烯胶乳 Polystyrene latex

触发分析的遮拦值 5%-20% Analysis in the trigger obscuration 5% -20%

获取时间 10 s Acquisition time 10 s

2样品制备 2 Sample preparation

- 制备0.1Q/。 - Preparation 0.1Q /. 的Span 80庚垸溶液(称取0.01 g Span 80粉末于20 ml烧瓶 Heptyl embankment of Span 80 solution (0.01 g Span 80 weighed powder in 20 ml flask

中,然后加入庚烷,控制最终的重量为10g), Then heptane was added to control the final weight of 10g),

- 称取约6 mg粉末于5 ml试管中, - Weigh about 6 mg powder in 5 ml test tube,

- 加入0.7 g含有0.P/Q Span 80的庚烷于该试管中, - addition of 0.7 g containing 0.P / Q Span heptane in the tube 80,

- 试管置于超声池中2分钟以彻底分散粉末。 - The tube was placed for 2 minutes to thoroughly disperse the powder ultrasonic bath.

3样品分析 3 Sample Analysis

清空储存于静止的液体-通路分散系统中的循环流体,然后加入庚垸。 Emptying the liquid stored in the stationary - circulating fluid passageway dispersion system, and then added heptane embankment. Hydro 2000SM转子的搅拌速度单位设置在2400 rev/min。 Hydro 2000SM speed stirring unit disposed in the rotor 2400 rev / min.

测试在前面提到的试验条件下开始: In the test under the test conditions mentioned earlier start:

-校正激光束, - correction of the laser beam,

-记录背景噪音。 - recorded background noise.

这些步骤之后操作员按照以下方式加入分析样品: These steps the operator after adding samples were analyzed as follows:

滴加稀释的样品(用Pasteur管),直到遮拦值为5%-20%,开始捕获。 Diluted sample was added dropwise (with a Pasteur pipe), until the obscuration value of 5% -20%, to begin capturing.

获得数据和D50有关,它是一个数值,即被分析目标中直径小于该值的占50%。 Obtaining related data and D50, which is a value smaller than the target diameter value i.e. 50% Analysis.

在三个不同样品中检测D50值三次,然后取其平均值。 D50 value detected three times in three different samples and averaged.

21b-Tl试验,其中微颗粒是水性分散体形式 21b-Tl trial, wherein the microparticles are in the form of an aqueous dispersion of

1设备和操作条件 <table>table see original document page 22</column></row> <table> 1 equipment and operating conditions <table> table see original document page 22 </ column> </ row> <table>

2样品制备 待分析样品加入到样品池中或者高散射样品也可以可选地用水再稀释。 Preparation of Sample 2 was added to the sample to be analyzed or highly scattering sample cell sample may also be optionally further diluted with water.

3样品分析 3 Sample Analysis

清空储存于静止的液体-通路分散系统中的循环流体,然后加入新鲜的去离子水。 Emptying the liquid stored in the stationary - circulating fluid passageway dispersion system, and fresh deionized water was added. Hydro 2000SM转子的搅拌速度单位设置在2400 rev/min。 Hydro 2000SM speed stirring unit disposed in the rotor 2400 rev / min.

测试在前面提到的试验条件下开始: In the test under the test conditions mentioned earlier start:

-校正激光束, - correction of the laser beam,

-记录背景噪音。 - recorded background noise.

这些步骤之后操作员按照以下方式加入分析样品: These steps the operator after adding samples were analyzed as follows:

滴加稀释的样品(用Pasteur管),直到遮拦值为5%-20%,开始捕获。 Diluted sample was added dropwise (with a Pasteur pipe), until the obscuration value of 5% -20%, to begin capturing.

获得数据和D50有关,它是一个数值,即被分析目标中直径小于该值的占50%。 Obtaining related data and D50, which is a value smaller than the target diameter value i.e. 50% Analysis.

在三个不同样品中检测D50值三次,然后取其平均值。 D50 value detected three times in three different samples and averaged. c-T2试验,其中微颗粒是有机溶剂中的分散体形式该试验和T1试验类似。 c-T2 test, a dispersion in which the microparticles in the form of an organic solvent and the test similar to Test T1. 然而,在本试验中,有必要将水替换成和分散液完全互溶的并且颗粒在其中不膨胀的溶剂。 However, in this test, it is necessary to replace the water and the dispersion and the particles are completely miscible in the solvent in which no swelling. 在多个实例中是使用了庚垸。 In various examples, it is the use of heptane embankment.

<table>table see original document page 23</column></row> <table>2样品制备 <Table> table see original document page 23 </ column> </ row> <table> 2 Sample Preparation

为制备待分析样品,400 pl待分析样品必须用600 jil庚垸在5 ml试管中稀释,然后该样品必须搅拌10秒(10士5)。 For the preparation of sample to be analyzed, 400 pl sample to be analyzed must be diluted in 5 ml test tubes with 600 jil heptyl embankment, the sample must then stirred for 10 seconds (10 ± 5).

3样品分析 3 Sample Analysis

清空储存于静止的液体-通路分散系统中的循环流体,然后加入新鲜的去离子水。 Emptying the liquid stored in the stationary - circulating fluid passageway dispersion system, and fresh deionized water was added. Hydro 2000SM转子的搅拌速度单位设置在2400 rev/min。 Hydro 2000SM speed stirring unit disposed in the rotor 2400 rev / min.

测试在前面提到的试验条件下开始: In the test under the test conditions mentioned earlier start:

-校正激光束, - correction of the laser beam,

-记录背景噪音。 - recorded background noise.

这些步骤之后操作员按照以下方式加入分析样品: These steps the operator after adding samples were analyzed as follows:

滴加稀释的样品(用Pasteur管),直到遮拦值为5%-20%,开始捕获。 Diluted sample was added dropwise (with a Pasteur pipe), until the obscuration value of 5% -20%, to begin capturing.

获得数据和D50有关,它是一个数值,即被分析目标中直径小于该值的占50%。 Obtaining related data and D50, which is a value smaller than the target diameter value i.e. 50% Analysis.

在三个不同样品中检测D50值三次,然后取其平均值。 D50 value detected three times in three different samples and averaged. 该特征由下面描述的DP1试验客观地定义。 The DP1 characterized by the test described below defined objectively. 粉末的"可分散性"DP1试验: Powder "dispersible" DP1 Test:

大约30 mg的颗粒粉末加入到带有隔板的3 ml烧瓶中。 About 30 mg of particles was added to the flask with 3 ml of the separator. 然后测定颗粒的精确重量(w,)。 Then determine the exact weight of the particles (w,). 用装有25G x 5/8 (0.5 x 16 mm)针头(例如BD Microlance 3型号)的1 ml注射器通过隔板将1 ml重构溶液加入到粉末中,然后该烧瓶手工间歇地搅拌15分钟。 Equipped with a 25G x 5/8 (0.5 x 16 mm) needle (e.g. Model BD Microlance 3) of a 1 ml syringe through the separator 1 ml of reconstituted solution was added to the powder, and then the flask was stirred by hand intermittently for 15 minutes. 烧瓶称重以确定所加入溶液的精确重量(W2)。 The flask was weighed to determine the exact weight of the solution (W2) was added. 最后,用装有25G针头的1 ml注射器(Braun Injeckt-F Luer 1 ml型,ref.9166017V)吸取全部悬浮液,然 Finally, all the suspension was suction a 1 ml syringe (Braun Injeckt-F Luer 1 ml type, ref.9166017V) equipped with 25G needle, then

后倾入到预先称皮重的另一个烧瓶中,检测所回收溶液的重量W3。 After pre-poured into another tared flask detecting the weight W3 of the solution recovered.

根据Tl或T2试验(取决于分散液体是水性溶剂或有机液体)测定分散体的质量方面(分散的倾向性、不透明性、溶液的视觉均一性)、通过25G针头注射的可能性、溶液的回收百分数:%=w3xl00/(Wl+w2),以及微颗粒的尺寸。 (Depending on the dispersion liquid is an aqueous liquid or an organic solvent) determining the quality of dispersion (dispersion tendency, opacity, visual uniformity solution) The test Tl or T2, recovered through 25G injection needle possibility solution percentage:% = w3xl00 / (Wl + w2), and the size of microparticles.

如果符合以下条件,则认为粉末在液体中的分散性质良好: If the following conditions are considered well dispersed in the liquid in the powder properties:

■所得悬浮液含有平均直径为2-40pm的微颗粒, "悬浮液的外观是均一的, "能够通过25G针头注射,以及•至少80%的悬浮液可被回收。 ■ The resulting suspension of microparticles having an average diameter of 2-40pm, "the appearance of the suspension is uniform," through 25G injection needle, and • at least 80% suspension can be recovered. 除特征(a)、 (b)和(c)外,根据本发明的微颗粒可以由它们在ST1试验和/或ST2试验中是稳定的事实客观地定义。 In addition to feature (a), (b) and (c), the microparticles according to the present invention may be stable in the fact that objectively defined and test ST1 / ST2 assay or by them.

ST1试验,用于测定干燥微颗粒的物理稳定性: ST1 test for determining the physical stability of the dried microparticles:

干燥雾化的粉末中的微颗粒的尺寸在粉末雾化后的一周内根据TO试验用激光颗粒尺寸检测仪测定。 Size of the micro particles in a dry atomized powder particle size measured by a laser detector within one week of the test powder atomizer according to TO. 然后颗粒置于3(TC烘箱中一周。"对照"样品在5"C放置。分散后按照和时间tO相同的条件再次测定尺寸。比较陈化前后的颗粒分布。 The particles were then placed in 3 (TC oven for one week. "Control" sample 5 "C is placed. After the dispersion was measured in accordance with the size of the time tO and the same condition again. Particle distribution compared before and after aging.

ST2试验,用于测定悬浮液中颗粒的胶体稳定性: ST2 test for determining the stability of the colloidal suspension of particles:

微颗粒粉末用磁力搅拌分散在分散液体中,其浓度为观察其稳定性的理想浓度,然后用中速磁力搅拌至少2h(t0检测)。 Stirred with a magnetic micro particles dispersed in a dispersion liquid, a concentration of the stability was observed over the concentration, and then stirred for at least 2h (t0 detection) speed with a magnetic force.

根据T1或T2试验在激光颗粒尺寸检测仪上根据分散介质(水性或有机)测定颗粒的尺寸。 The measured particle size dispersion medium (aqueous or organic) particle size on the laser detector according to Test T1 or T2.

悬浮液然后于5T:下放置7天。 The suspension was then to 5T: placing at 7 days. 沉淀的颗粒搅拌几分钟分散(直至所得悬浮液 The precipitated particles dispersed stirred for several minutes (until the resulting suspension

视觉上是均一的)。 Visually uniform). 分散后按照和时间to相同的条件再次测定尺寸。 Measured according to size and time again to the same dispersing conditions. 聚合物PO Polymer PO

根据本发明的聚合物PO是带有疏水性基团HG和亲水性基团[优选部分电离的可电离基团IG]的水溶性生物可降解聚合物。 Water-soluble biological polymer according to the present invention, PO is a group having a hydrophobic group and a hydrophilic group HG [preferably partially ionized ionizable groups the IG] degradable polymers. 和链的剩余部分相比,疏水性基团HG的数目可以少一些,并且可以从侧面连接到链上或者插在链中,并且可以随机分布(无规共聚物)或者以序列或者接枝的形式分布(嵌段共聚物或连续共聚物)。 And the remainder of the chain as compared to the number of hydrophobic groups HG may be less, and may be connected to the chain, or from the side is inserted in the chain, and may be distributed randomly (random copolymer) or in a sequence or grafted distributed fashion (continuous copolymer or a block copolymer).

根据本发明一个优选的实施方式,聚合物PO是两性分子(共)聚氨基酸。 According to a preferred embodiment of the invention, the polymer PO is amphiphilic (co) polyamino acids. PO的这种选择提供了和蛋白质/肽类型AP优良的相容性。 This provides a choice of PO and protein / peptide type AP excellent compatibility. 因此能够极大地简化蛋白质/肽类型AP和聚合物PO的起始溶液或者悬浮液的组成。 It is possible to greatly simplify the composition of a protein / peptide and the type of polymer PO AP starting solution or suspension. 这种起始溶液或悬浮液可以仅含有在水性和/或有机相中的AP和PO。 This starting solution or suspension may contain only an aqueous and / or organic phase of the AP and PO. 对于这样的起始溶液或悬浮液,不含有其它成分使得能够在雾化前过滤(通过0.2 pm的滤膜),这使得能够在无菌条件下操作后者。 For such starting solution or suspension, containing no other ingredients enables filtering (0.2 pm filter) prior to atomization, which makes it possible to operate the latter under sterile conditions.

根据本发明一个优选的实施方式中,PO选自两性分子(共)聚氨基酸及其混合物。 According to a preferred embodiment of the present invention, PO is selected from amphiphilic molecules (co) poly amino acids and mixtures thereof.

优选地,根据本发明的聚氨基酸是低聚物或均聚物,含有谷氨酸或天冬氨酸重复残基,或共聚物体,含有这两种类型氨基酸残基的混合物。 Preferably, the polyamino acids according to the present invention are oligomers or homopolymers comprising glutamic or aspartic acid repeat residues, or copolymer thereof, a mixture containing these two types of amino acid residues. 这些聚合物中所述的残基是具有D或L或D/L构型的氨基酸,就谷氨酸或谷氨酸残基来说,其通过它们的a或Y位结合,就天冬氨酸或天冬氨酸残基来说,其通过它们的a或(3位结合。 These residues are polymers having the D or L or D / L-configuration amino acids, glutamic acid or glutamic acid residue on it, which is incorporated by their position a or Y, it aspartyl acid or aspartic acid residues for which they are joined by a or (3.

根据本发明一个特别优选的实施方式,聚合物PO是由天冬氨酸残基和/或谷氨酸残基形成的聚氨基酸,至少一部分这些残基中带有包含至少一个疏水性基团HG的接枝。 According to a particularly preferred embodiment of the present invention, the polymer PO is a polyamino acid formed by aspartic acid residues and / or glutamic acid residues, at least a portion of these residues having comprising at least one hydrophobic group HG the graft. PCT专利申请WO-A-00/30618中特别描述了这些类型的聚氨基酸。 PCT patent application WO-A-00 medium / 30618 describes these particular types of polyamino acids.

根据第一种可能性,所述PO由下列通式(I)(基团-COOR?包括羧基和113之间的键是离子键-COO'+R3的形式)定义:<formula>formula see original document page 26</formula>其中: According to a first possibility, the PO by the following general formula (I) (including a carboxyl group -COOR 113 and the bond between the ionic bond -COO '+ R3 form?) Is defined: <formula> formula see original document page 26 </ formula> where:

■ R1代表H、 C2到C1Q的直链垸基或C3到C1Q的支链垸基、苄基和-Rt[HG],或者 ■ R1 representative of H, C2 to C1Q straight-chain alkyl group or branched C3 to C1Q chain alkyl, benzyl, and -Rt [HG], or

■ NHI^是末端氨基酸残基; ■ NHI ^ is a terminal amino acid residue;

■ R2代表H、 C2到C1()的直链酰基或C3到C1()的支链酰基、焦谷氨酸根基团或-R、[HG]; ■ R2 Representative H, a C1 to C2 () a straight-chain acyl group or a C1 to C3 () branched chain acyl, or pyroglutamic acid radicals -R, [HG];

■ RS是H,或者 ■ RS is H, or

■ +113优选自包括以下的组: ■ +113 preferably selected from the group consisting of:

- 金属阳离子,优选自包括钠、钾、钙和镁的亚组, - metal cation, preferably from the group consisting of sodium, potassium, calcium and magnesium, subgroup,

- 有机阳离子,优选自包括以下的亚组: - an organic cation, preferably from the subgroup comprising:

• 基于胺的阳离子, • cations based on amine,

• 基于低聚胺的阳离子, • cations based on oligomeric amine

• 基于聚胺的阳离子(特别优选聚乙烯亚胺), • cations based on polyamine (polyethyleneimine is particularly preferred),

• 基于氨基酸的阳离子,优选自包括基于赖氨酸或精氨酸的阳离子的组, • cations based on amino acid, preferably selected from the group consisting of cations based on lysine or arginine, and

- 或者阳离子聚氨基酸,优选自包括聚赖氨酸和低聚赖氨酸的亚组; - or cationic polyamino acids, preferably from the subgroup comprising polylysine and oligomeric lysine;

■ W代表直接键或基于1到4个氨基酸残基的"间隔子"; ■ W represents a direct bond or a 1 to 4 based on the amino acid residues of the "spacer";

■ 独立地,A代表-(:112-基团(天冬氨酸残基)或-0^2-(3112-基团(谷氨酸残基); ■ independently, A represents - (: 112- group (aspartic acid residue), or -0 ^ 2- (3112- groups (glutamic acid residues);

■ n/(n+m)是摩尔接枝率,在pH为7, 25'C时溶于水中时PO的摩尔接枝率值足够低,以形成PO亚微米尺寸颗粒的胶体悬浮液;n/(n+m)优选为1 -25 mol% ,更优选1 -15 mol%;■ n+m是聚合度,其为10-1000,优选50-300; ■ n / (n + m) is the molar grafting rate, at pH 7, when dissolved in water, at 25'C value PO molar grafting rate is sufficiently low, to form a colloidal suspension of submicron sized particles of PO; n- / (n + m) is preferably from 1 -25 mol%, more preferably 1 -15 mol%; ■ n + m is the degree of polymerization, which is 10-1000, preferably 50-300;

■ HG代表含有6-30个碳原子的疏水性基团。 ■ HG Representative hydrophobic group containing 6-30 carbon atoms group. 在本发明一个优选的实施方式中,所述疏水性基团HG选自含有以下类型 In a preferred embodiment of the present invention, the hydrophobic group is selected from the group comprising the following type HG

的烷氧基的组:一OCH2(CH2-CH2)3.8~CH3、油烯基、生育酚基或胆甾醇基,并且W代表一个单键。 Alkoxy group: a OCH2 (CH2-CH2) 3.8 ~ CH3, oleyl, tocopheryl or cholesteryl group, and W represents a single bond.

根据第二种可能性,PO对应于以下通式(II)、 (ni)和(IV)(基团-COORS包括羧基和W之间的键是离子键-COO'+W的形式)中的一种: According to a second possibility, PO corresponding to the following general formula (II), (ni) and (IV) (-COORS group include a carboxyl group and the bond between W is an ionic bond -COO '+ W form) in the one of:

COOR3' ,COOr3' COOR3 ', COOr3'

OH OH

(IV) (IV)

其中: among them:

■ HG代表含有6-30个碳原子的疏水性基团; Representative ■ HG containing a hydrophobic group from 6 to 30 carbon atoms;

■ R^是直链的二价CVC6亚烃基链; ■ R ^ is a divalent linear alkylene chain CVC6;

■ R3'是H,或者 ■ R3 'is H, or

■ +113'优选自包括以下的组: ■ +113 'is preferably selected from the group consisting of:

- 金属阳离子,优选自包括钠、钾、钙和镁的亚组, - metal cation, preferably from the group consisting of sodium, potassium, calcium and magnesium, subgroup,

- 有机阳离子,优选自包括以下的亚组: - an organic cation, preferably from the subgroup comprising:

• 基于胺的阳离子, • cations based on amine,

• 基于低聚胺的阳离子, • cations based on oligomeric amine

• 基于聚胺的阳离子(特别优选聚乙烯亚胺), • cations based on polyamine (polyethyleneimine is particularly preferred),

• 基于氨基酸的阳离子,优选自包括基于赖氨酸或精氨酸的阳离子的组, • cations based on amino acid, preferably selected from the group consisting of cations based on lysine or arginine, and

27- 或者阳离子聚氨基酸,优选自包括聚赖氨酸和低聚赖 27- or cationic polyamino acids, polylysine and preferably from the group consisting oligomeric Lai

氨酸的亚组; Acid subgroup;

■ R5G是直链的二价CVQ亚烃基链,其中一个或两个优选位于R5G每个末端的亚甲基单元可以独立地被-O-或-NH代替; ■ R5G is a divalent linear alkylene chain CVQ, wherein one or two preferably located at each end of the R5G methylene units may independently be replaced by -O- or -NH;

■ R"代表直接键或基于1到4个氮基酸残基的间隔子; ■ R "represents a direct bond or a spacer based on 1 to 4 nitrogen atom of the amino acid residues;

■ 独立地,A代表-CH2-基团(天冬氨酸残基)或-CH2-CH2-基团(谷氨酸残基); ■ independently, A represents a group -CH2- (aspartic acid residue) or a group -CH2-CH2- (glutamic acid residues);

■ n,+m'或n"是聚合度,其为IO-IOOO,优选50-300。 根据一个优选的可供选择的形式,W代表一个单键。 ■ n, + m 'or n "is the degree of polymerization, which is IO-IOOO, preferably 50-300. According to a preferred form of alternative, W represents a single bond.

根据第三种可能性,PO是基本上中性的共聚羟基烷基谷氨酰胺(优选地垸基是乙基),其含有多个相同或互不相同的疏水性基团(HG)侧基。 According to a third possibility, PO is substantially neutral copolyhydroxyalkylglutamine (preferably alkyl with ethyl), comprising a plurality of same or different from the hydrophobic groups (HG) side group . 所述共聚羟基烷基谷氨酰胺还带有羟基烷基胺基团。 The copolyhydroxyalkylglutamine further amine group with a hydroxyl group. 这些羟基垸基胺基团优选通过酰胺键结合到所述共聚物。 These hydroxyl alkyl with amine groups preferably incorporated into the copolymer via an amide bond. 可用于酰胺化谷氨酸残基的羧基并用于形成所述共聚羟基烷基谷氨酰胺的这些羟基垸基胺基团可以相同或者互不相同,例如选自以下基团:2-羟基乙基胺、3-羟基丙胺、2,3-二羟基丙胺、三(羟基甲基)氨基甲垸和6-羟基己胺。 It can be used for the amidation of a carboxyl group of glutamic acid residues and used to form the copolyhydroxyalkylglutamine embankment of these hydroxyl groups or amine groups may be the same different from each other, for example, selected from the group: 2-hydroxyethyl amine, 3-hydroxy-propylamine, 2,3-dihydroxy-propylamine, tris (hydroxymethyl) carbamoyl embankment and 6-hydroxyhexyl amine.

优选地,所述疏水性基团HG中的至少一个包括在疏水性接枝物中,所述疏水性接枝物含有至少一个间隔接合点(或单元)(间隔子),其能将所述疏水性基团HG连接到共聚谷氨酸的链上。 Preferably, the hydrophobic group HG comprises at least one hydrophobic graft, the hydrophobic grafts containing at least one engagement point interval (or cell) (spacer), which is capable of the HG hydrophobic group attached to the copolymer chain of glutamic acid. 该接合点可以包括,例如至少一个直接的共价键或至少一个酰胺键或至少一个酯键。 The joint may comprise, for example, at least one direct covalent bond or an amide bond or at least one of the at least one ester bond. 例如,该接合点可以尤其属于以下组的类型:氨基酸残基、氨基醇的衍生物、聚胺的衍生物(例如二胺)、多元醇的衍生物(例如二醇)和多羟基酸的衍生物。 For example, the joint may in particular belong to the group of the type: amino acid residues, amino alcohol derivatives, polyamine derivatives (e.g. diamines), derivatives derived from polyols (e.g. glycols) and polyhydroxy acids thereof. 所述HG和共聚谷氨酸或聚垸基谷氨酰胺链的接枝可以包括使用能够键合到所述共聚谷氨酸或共聚羟基烷基谷氨酰胺链的HG 前体。 HG and the polyethylene graft copolymer alkyl with glutamine or glutamic acid may include the use of chain capable of bonding to the copolymerization of glutamic acid or glutamine copolymerizable hydroxyalkyl front HG chain thereof. 所述HG的前体实际上没有限制,选自包括醇类和胺类的组,这些化合物能够容易地被本领域的技术人员功能化。 The precursor of HG virtually no limitation, selected from the group comprising alcohols and amines, these compounds can easily be functionalized those skilled in the art. 关于这种共聚羟基垸基谷氨酰胺(优选的垸基是乙基)的细节参见FR-A-2,881,140。 Details on this comonomer hydroxy alkyl with glutamine (preferably alkyl with ethyl) See FR-A-2,881,140.

根据一个优选的可供选择的形式,尤其根据上述三种可能性的至少一种, 所述PO的所有或部分疏水性基团HG独立地选自包括以下基团的组: According to a preferred alternative form, in particular according to at least one of the three possibilities, the PO of all or part of the hydrophobic groups HG independently selected from the group consisting of the following groups:

■ 含有6到30个碳原子的直链或支链烷氧基并且其可以含有至少一个杂原子(优选O、 N或S)和/或至少一个不饱和键,■ 含有6到30个碳原子的垸氧基,其具有一个或多个退火 ■ containing from 6 to 30 carbon atoms, a straight-chain or branched alkoxy group and which may contain at least one heteroatom (preferably O, N or S) and / or at least one unsaturated bond, ■ containing from 6 to 30 carbon atoms the embankment group having one or more annealing

的环烷基(cycloalkyles annel6s),并且可选地,含有至少一个不饱和键或至少一个杂原子(优选O、 N或S), Cycloalkyl (cycloalkyles annel6s), and optionally containing at least one unsaturated bond or at least one heteroatom (preferably O, N or S),

■ 含有7到30个碳原子的烷氧基芳基或芳氧基垸基,其含有至少一个不饱和键或至少一个杂原子(优选O、 n或S)。 ■ an aryl group an alkoxy group or aryloxy alkyl with 7-30 carbon atoms, containing at least one unsaturated bond or at least one heteroatom (preferably O, n, or S).

根据一个优选的可供选择的形式,尤其根据上述三种可能性的至少一种, 所述疏水性基团HG选自含有以下类型的垸氧基的组:-OCH2(CH2-CH2)3.8~€H3、 油烯基、生育酚基或胆甾醇基,并且R"代表一个单键。 According to a preferred alternative form, in particular according to at least one of the above three possibilities, the hydrophobic group HG is selected from the group comprising the following types of embankment group: -OCH2 (CH2-CH2) 3.8 ~ € H3 of, oleyl, tocopheryl or cholesteryl group, and R "represents a single bond.

根据另一个优选的可供选择的形式,尤其根据上述三种可能性的至少一种,所述PO的n个HG基团各自独立地代表下式的单价基团: According to another preferred alternative form, in particular according to at least one of the n HG of the PO each independently represent a group of the three possibilities monovalent radical of the formula:

(HG) (HG)

其中: among them:

- RS代表甲基(丙氨酸)、异丙基(缬氨酸)、异丁基(亮氨酸)、仲丁基(异亮氨酸)或苄基(苯丙氨酸); - RS represents a methyl (alanine), isopropyl (valine), isobutyl (leucine), sec-butyl (isoleucine) or benzyl (phenylalanine);

- W代表含有6到30个碳原子的疏水性基团; - W represents an 6-30 hydrophobic group carbon atoms;

- l为0到6。 - l is 0-6.

根据本发明的一个显著特征,所述PO的所有或部分疏水性基团R^虫立地选 According to a significant feature of the present invention, all or part of the PO is a hydrophobic group selected from R ^ insect habitats

自包括以下基团的组: From the group consisting of the following groups:

■ 含有6到30个碳原子的直链或支链烷氧基并且其可以含有至少一个杂原子(优选O、 N或S)或至少一个不饱和键, ■ containing from 6 to 30 carbon atoms, a straight-chain or branched alkoxy group and which may contain at least one heteroatom (preferably O, N or S) or at least one unsaturated bond,

■ 含有6到30个碳原子的烷氧基,其具有一个或多个退火的环烷基(cycloalkyles annel6s),并且可选地,含有至少一个不饱和键或至少一个杂原子(优选O、 N或S), ■ containing 6 to 30 carbon atoms, an alkoxy group having one or more annealing cycloalkyl (cycloalkyles annel6s), and optionally containing at least one unsaturated bond or at least one heteroatom (preferably O, N or S),

■ 含有7到30个碳原子的烷氧基芳基或芳氧基烷基,并且含有至少一个不饱和键或至少一个杂原子(优选O、 N或S)。 ■ an aryl group an alkoxy group or aryloxy group having 7 to 30 carbon atoms, and containing at least one unsaturated bond or at least one heteroatom (preferably O, N or S).

实际上没有限制,所述PO的接枝物的疏水性基团W选自含有以下类型的烷氧基的组:-OCH2(CH2-CH2)3-8~CH3、油烯基、生育酚基或胆甾醇基,并且W代表一个单键。 Virtually no limitation, PO hydrophobic group W is selected from a graft of the type of alkoxy group: -OCH2 (CH2-CH2) 3-8 ~ CH3, oleyl, tocopheryl or cholesteryl, and W represents a single bond.

优选地,聚氨基酸的主链是: Preferably, the main polyamino acid chain are:

► aL-谷氨酸根或aL-谷氨酸均聚物; ► aL- aL-glutamate or glutamic acid homopolymer;

► aL-天冬氨酸根或aL-天冬氨酸均聚物; ► aL- aL-aspartate or aspartic acid homopolymer;

► 或aL-天冬氨酸根/aL-谷氨酸根或aL天冬氨酸/aL-谷氨酸共聚物。 ► aL-aspartate or / aL- aL glutamate or aspartate / aL- acid copolymer. 优选地,PO的聚氨基酸主链的天冬氨酸或谷氨酸残基的分布使得到的聚合 Preferably, PO or the distribution of aspartic acid glutamic acid residues of the polyamino acid backbone resulting polymer

物是无序的或嵌段型的或复块型的。 It was disordered or block-type or complex-type blocks.

根据另一种定义的方法,PO的摩尔质量在2000-100,000g/mo1,优选5000-40,000g/mol。 According to another method defined by the molar mass of the PO 2000-100,000g / mo1, preferably 5000-40,000g / mol.

根据一个可供选择的形式,PO带有至少一个聚二醇类接枝,其与谷氨酸根或天冬氨酸根残基键合。 According to an alternative form, PO with at least one grafted polyethylene glycols, which glutamate or aspartate residues bonded.

优选地,该聚二醇类接枝为下式(V)所示类型: Preferably, the polyethylene glycol is a graft type the following formula (V):

在实践中,聚二醇是例如聚乙二醇。 In practice, for example, polyethylene glycol is polyethylene glycol.

根据本发明,理想的聚二醇接枝的摩尔百分率为1到30%。 According to the present invention, the molar percentage of polyalkylene glycol grafting is preferably 1 to 30%.

此外,聚氨基酸PO在下述方面非常有价值,即它们以可调节的接枝率分散在pH:7.4(例如有磷酸盐缓冲液)的水中,形成胶态悬浮液。 Further, polyamino acids PO is very valuable in the following, i.e., they are adjustable grafting rate was dispersed in pH: 7.4 (for example, phosphate buffer) water, to form a colloidal suspension.

而且,活性成分AP,例如蛋白质、肽或小分子,可以自发地与包括这些聚氨基酸PO的纳米颗粒结合在一起。 Further, the AP active ingredient, such as proteins, peptides or small molecules, may spontaneously combine with nanoparticles comprising these polyamino acids PO's.

应该理解的是,包含可电离功能基团的PO可以是中性的(例如-COOH)或电离的(例如-COCT),视pH和组合物而定。 It should be understood, PO containing ionizable functional groups may be neutral (e.g. -COOH) or ionized (e.g. -COCT), depending on the composition and pH dependent. 因此,在含水相中的溶解度直接取决于可电离功能基团的比例和pH。 Thus, the solubility of the aqueous phase and the pH is directly dependent on the proportion of ionizable functional groups. 当水性溶液中有羧基功能基时,抗衡离子可以是 When the aqueous solution with a carboxyl functional group, the counter ion may be

其中: among them:

R'"代表直接键或基于l到4个氨基酸残基的"间隔 R ' "represents a direct bond or based l to 4 amino acid residues" spacer

X是选自氧、氮和硫的杂原子; W和RS独立地代表H或直链的d到C4烷基; n"'为10到1000,优选50到300。 X is a hetero atom selected from oxygen, nitrogen and sulfur; W is and RS independently represent H or linear d to C4 alkyl group; n " 'is 10 to 1000, preferably 50-300.

30金属阳离子(例如钠、钙或镁),或有机阳离子(例如三乙醇胺,三羟甲基氨基甲垸的质子形式),或聚胺(例如聚乙烯亚胺)。 30 metal cation (e.g., sodium, calcium or magnesium), or an organic cation (such as triethanolamine, tris (hydroxymethyl) carbamoyl embankment proton form), or polyamines (e.g. polyethyleneimine).

聚氨基酸类的PO可以通过例如为本领域技术人员所知的方法得到。 The polyamino acids PO can be obtained, for example, by methods known to those skilled in the art. 无序聚氨基酸可以通过普通的偶联反应,将疏水基(预先被"间隔子"功能化)直接连接到 Poly amino acids may be disordered by a general coupling reaction, directly connected to the hydrophobic group (previously "spacer" functionalized)

聚合物上而得到。 The polymer obtained. 嵌段或复块聚氨基酸PO可以通过相应N-羧基氨基酸酐(NCA) Polyamino acid block or a complex block by the corresponding N- PO-carboxy amino acid anhydride (NCA)

的连续聚合得到。 The continuous polymerization.

例如,共聚谷氨酸根、共聚天冬氨酸根聚氨基酸或嵌段、复块或无序谷氨酸根/天冬氨酸根共聚物由常规方法制备。 For example, co-glutamate, aspartate copolymer polyamino acid or a block, random or block complex glutamate / aspartate copolymer is prepared by conventional methods.

为得到a型聚氨基酸,最常用的方法是基于N-羧基氨基酸酐(NCA)的聚合, 对该方法例如在"Bi叩olymers, 1976, 15, 1869"的文章以及HRKricheldorf所作的题为"a-amino acid N隱carboxy anhydrides and related heterocycles ,,, Springer Verlag (1987)的书中有描述。当侧链具有羧酸功能时,NCA衍生物优选为NCA-O-Me、 NCA-O-Et或NCA-O-Bz衍生物(Me二甲基,Et二乙基,Bz二节基)。然后将聚合物在适当的条件下水解,以得到它的酸形式聚合物。这些方法基于申请人的在专利FR-A-2801226中的描述。本发明可使用的某些聚合物,例如,分子量不同的聚(aL-天冬氨酸)、聚(aL-谷氨酸)、聚(aD-谷氨酸)和聚(?L-谷氨酸) 类,均可以商购获得。a,p型的聚天冬氨酸经天冬氨酸的縮合(以得到聚琥珀酰亚胺),然后是碱水解得至U(参考Tomida等人的Polymer, 1997, 38, 4733-36)。 To obtain a polylactic acid, the most commonly used method is based on N--carboxy amino acid anhydride (NCA) polymerization, for example, in the method "Bi knock olymers, 1976, 15, 1869" and the article entitled HRKricheldorf made "a -amino acid N implicit carboxy anhydrides and related heterocycles ,,, Springer Verlag (1987) is described in the book. when a side chain having a carboxylic acid function, NCA derivatives are preferably NCA-O-Me, NCA-O-Et or NCA-O-Bz derivatives (Me dimethyl, Et diethyl, Bz two-yl). the polymer is then hydrolyzed under suitable conditions to obtain its acid form polymers. these methods are based on applicants' described in the patent FR-a-2801226 in certain polymers of the present invention can be used, for example, different molecular weights poly (aL-aspartic acid), poly (aL-glutamic acid), poly (aD-Valley acid) and poly (? L-glutamic acid) s, poly-aspartic acid can be commercially available by the condensation of aspartic acid .a, p-type (to give a polysuccinimide) followed by alkaline hydrolysis to obtain U (refer to Tomida et al., Polymer, 1997, 38, 4733-36).

接枝物与聚合物的酸功能基的偶合通过聚氨基酸在碳二亚胺作为偶联剂、 以及可选的催化剂例如4-二甲基氨基吡啶的存在下,于适当的溶剂中容易地进行反应,溶剂例如为二甲基甲酰胺(DMF)、 N-甲基吡咯垸酮(NMP)或二甲亚砜(DMSO)。 Coupling the acid functional groups of the polymer by graft polymerization of amino acids in a carbodiimide as coupling agent, and optionally a catalyst such as 4-dimethylaminopyridine, in a suitable solvent readily reaction, a solvent such as dimethylformamide (DMF), N- methylpyrrole embankment ketone (NMP) or dimethyl sulfoxide (DMSO). 碳二亚胺例如是二环己基碳二亚胺或二异丙基碳二亚胺。 Carbodiimide, for example dicyclohexyl carbodiimide or diisopropyl carbodiimide. 接枝率由组分和反应物的化学当量控制,或通过反应时间控制。 Chemical equivalent component and the grafting rate of the reaction was controlled, or controlled by the reaction time. 由"间隔子"功能化的疏水性接枝通过常规的肽偶合或通过在酸催化之下直接縮合得到。 Hydrophobic graft "spacer" functionalized by conventional peptide coupling or by direct condensation obtained under acid catalysis. 这些技术均为所属领域的技术人员所熟知。 These techniques are all ordinary skill in the art in the art.

由预先与疏水性接枝合成的NCA衍生物来合成嵌段或复块共聚物。 The NCA derivatives previously synthesized with the hydrophobic graft or block of synthesized complex block copolymer. 例如, 疏水NCA衍生物与NCA-O-Bz进行共聚,然后通过水解选择性除去节基。 For example, the hydrophobic NCA derivative with NCA-O-Bz copolymerization, and then selectively removed by hydrolysis section group.

活性成分PA(s) Active ingredient PA (s)

关于AP,优选自以下组:蛋白质、糖蛋白、与一个或多个聚二醇链结合的蛋白质[优选聚乙二醇(PEG):"聚乙二醇化的蛋白质"]、肽、聚糖、脂糖、寡聚核苷酸、多核苷酸及其混合物,并且特别优选自以下亚组:血红蛋白、催产素、后叶加压素、促肾上腺皮质激素、表皮生长因子、血小板源性生长因子(PDGF)、造血刺激因子及其混合物、VIII因子和IX因子、血色素、细胞色素、 On the AP, preferably selected from the group consisting of: a protein, glycoprotein, with one or more polyethylene glycol chain binding protein [preferably polyethylene glycol (PEG): "PEGylated protein"], a peptide, glycan, liposaccharides, oligonucleotides, polynucleotides, and mixtures thereof, and particularly preferably from the following subgroup: hemoglobin, oxytocin, vasopressin, adrenocorticotropic hormone, epidermal growth factor, platelet derived growth factor ( PDGF), and mixtures thereof, hematopoiesis-stimulating factors, VIII and factor factor IX, hemoglobin, cytochrome,

白蛋白、泌乳刺激素、促黄体素释放素、黄体激素释放激素(LHRH)、 LHRH拮抗剂、LHRH竞争剂、人、猪或牛生长因子(GH)、生长激素释放因子、胰岛素、 生长激素抑制素、胰高血糖素、白介素或其混合物(IL-2、 IL-H、 IL-12)、 a-、卩-或Y-干扰素、胃泌素、四肽胃泌素、五肽胃泌素、尿抑胃素、分泌素、降钙素、 脑啡肽、内吗啡肽、血管紧张素、促甲状腺素释放激素(TRH)、肿瘤坏死因子(TNF)、神经生长因子(NGF)、粒细胞集落刺激因子(G-CSF)、粒细胞巨噬细胞集落刺激因子(GM-CSF)、巨噬细胞集落刺激因子(M-CSF)、肝素酶、骨成形素蛋白(BMP)、 hANP、类高血糖素样肽(GLP-1)、 VEG-F、重组乙型肝炎表面抗原(rHBsAg)、肾素、细胞因子、缓激肽、杆菌肽、多粘菌素、粘菌素、短杆菌酪肽、短杆菌肽、环孢菌素类,以及酶、细胞因子、抗体、抗原和疫苗的合成类似物和药学活性改 Albumin, prolactin, luteinizing hormone-releasing hormone, luteinizing hormone releasing hormone (LHRH), LHRH antagonists, of LHRH competitor, human, porcine or bovine growth factor (GH), growth hormone releasing factor, insulin, somatostatin hormone, glucagon, interleukin or mixtures thereof (IL-2, IL-H, IL-12), a-, Jie - Y- or interferon, gastrin, gastrin tetrapeptide, pentapeptide gastrin Su, gastric inhibitory urinary hormone, secretin, calcitonin, enkephalins, endomorphin, angiotensin, thyrotropin releasing hormone (TRH), tumor necrosis factor (of TNF), nerve growth factor (NGF), granulocyte colony stimulating factor (G-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (M-CSF), heparinase, bone morphogen protein (BMP), hANP, glucagon-like peptide (GLP-1), VEG-F, recombinant hepatitis B surface antigen (of rHBsAg), renin, cytokines, bradykinin, bacitracin, polymyxin, colistin, gramicidin casein peptides, gramicidin, cyclosporin, and enzymes, synthetic analogs of cytokines, antibodies, antigens and vaccines and pharmaceutically active modified 性物和片段。 Sexual and fragments thereof.

根据一个可供选择的形式,活性成分是疏水或亲水的,可选地离子的,"小的"有机分子,其类型属于蒽环霉素、紫杉类或喜树碱家族,或是属于肽(如亮丙瑞林或环孢子菌素)家族以及它们的混合物。 According to an alternative form, the active ingredient is hydrophilic or hydrophobic, optionally of ions, "small" organic molecule type is an anthracycline, a taxane or a camptothecin family, belonging or peptides (e.g., leuprolide or cyclosporin) family, and mixtures thereof.

本发明的内容中,"小"分子特别指非蛋白小分子,例如没有氨基酸的小分子。 In the context of the present invention, "small" molecule refers particularly to non-protein small molecule, such as a small molecule without amino acids.

根据另一个可供选择的形式,所述活性成分优选自以下活性物质家族的至少一种:治疗酗酒的药物、治疗阿尔茨海默病毒药物、麻醉药、治疗肢端肥大症的药物、镇痛药物、平喘药、治疗过敏的药物、抗肿瘤药、抗炎药、抗凝血剂和抗血栓形成剂、抗惊厥药、抗癫痫药、抗糖尿病药、止吐药、抗青光眼药、抗组胺药、抗感染药、抗生素、抗真菌药、抗病毒药、抗帕金森病药、抗胆碱药、止咳药、碳酸酐酶抑制剂、心血管药物、降血脂药、抗心律失常药、 血管扩张药、抗心绞痛药、抗高血压药、血管保护剂、胆碱酯酶抑制剂、治疗中枢神经系统失调的药物、中枢神经系统刺激剂、避孕药、生殖促进药、分娩促进药和抑制剂、治疗粘液粘稠病的药物、多巴胺受体激动剂、治疗子宫内膜异位的药物、治疗勃起障碍的药物、治疗生育的药物、治疗胃肠道疾 According to another alternative form, the active ingredient is preferably selected from the following families of active substances at least one of: drug therapy of alcoholism, drugs for treating Alzheimer virus, anesthetics, acromegaly therapy drugs, analgesic drugs, antiasthmatic drugs, allergy therapeutic drugs, antineoplastic agents, anti-inflammatory drugs, anticoagulants and antithrombotic agents, anticonvulsants, antiepileptics, anti-diabetic, anti-emetics, anti-glaucoma agents, anti- histamine drugs, anti-infectives, antibiotics, antifungal agents, antiviral agents, antiparkinsonian agents, anticholinergic agents, antitussives, carbonic anhydrase inhibitors, cardiovascular drugs, hypolipidemic drugs, antiarrhythmic agents, vasodilators agents, anti-anginal agents, antihypertensive agents, vascular protective agents, cholinesterase inhibitors, treatment of CNS disorder drugs, central nervous system stimulants, contraceptives, reproductive medicine to promote, facilitate drug delivery and inhibitors, Drug treatment of mucoviscidosis, dopamine receptor agonists, endometriosis drugs, erectile dysfunction drugs, the treatment of fertility drugs, the treatment of gastrointestinal diseases 病的药物、免疫调节剂和免疫抑制剂、治疗记忆疾病的药物、抗偏头痛药、肌松药、 Disease drugs, immunomodulators and immunosuppressants, a pharmaceutical treating memory disorders, antimigraine agents, muscle relaxants,

32核苷类似物、治疗骨质疏松的药物、拟副交感神经药、前列腺素、心理治疗药、镇静药、催眠和镇定药、安定药、抗焦虑药、精神振奋药、抗抑郁药、治疗皮肤病的药物、留体和激素、安非他明、减肥药、非镇痛型止痛药、巴比妥类、苯并二氮杂卓类、放松药、精神药品以及任何这些产品的联合使用。 32 nucleoside analogues, drugs for treating osteoporosis, parasympathomimetics, prostaglandins, psychotherapeutic agents, sedatives, tranquilizers and hypnotic, tranquilizers, anti-anxiety drugs, spirits, antidepressants, skin treatment disease drugs, stay and hormones, amphetamines, diet pills, non-type analgesic painkillers, barbiturates, benzodiazepines class, relax drugs, psychotropic substances and any joint use of these products.

从定量的角度看,特别优选不与微米尺寸的颗粒结合的AP[非结合AP]的重量百分率为以下数值: From a quantitative point of view, particularly preferably not bind the AP with the micron sized particles [non-bound the AP] The weight percentage of the following values:

o [非结合AP]^1, o [non-bound AP] ^ 1,

o优选[非结合AP]S0.5。 o preferably [non-bound AP] S0.5.

本发明的第二方面:通过雾化含有AP的PO溶液或胶体悬浮液生产微颗粒的 A second aspect of the present invention: by atomization of AP containing solution or a colloidal suspension of PO produced microparticles

方法 method

根据本发明的方法一个优选地方式,用于雾化的溶液或胶体悬浮液中存在 The method according to the present invention a preferred embodiment, for atomizing a solution or a colloidal suspension in the presence of

的PO至少部分为PO纳米颗粒的形式,所述纳米颗粒以T1试验测定的尺寸小于 PO is in the form of at least a portion of PO nanoparticles, the nanoparticles size is less than T1 experimentally determined

500nm,优选为10-300nm,特别优选IO-IOO nm。 500nm, preferably 10-300nm, particularly preferably IO-IOO nm.

优选地,待雾化的溶液或胶体悬浮液中PO的浓度为,例如5 mg/ml-100 Preferably, the concentration to be atomized solution or a colloidal suspension of PO, such as 5 mg / ml-100

mg/ml , 优选10mg/ml-40 mg/ml 。 mg / ml, preferably 10mg / ml-40 mg / ml.

本发明的上下文中,AP/PO结合物在雾化步骤之前和/或期间产生。 The context of the present invention, AP / PO conjugate produced before the atomization step, and / or period. 对于这种结合物,PO和/或AP可以是液体悬浮液形式中的固体形式(优选粉末)。 For this combination, PO and / or AP may be in solid form in a liquid suspension form (preferably a powder).

在雾化步骤前使一种或多种AP和PO相结合的技术尤其在专利申请WO-A-00/30618中有描述。 Before the step of atomizing AP and one or more techniques of combining PO especially in patent application WO-A-00/30618 is described. 这些技术例如是将PO的胶体悬浮液和AP的溶液或悬浮液混合。 These techniques are the colloidal suspension of PO and the AP, for example, mixing a solution or suspension. 根据一种可供选择的方式,粉末形式的AP和PO悬浮液相混合。 According to an alternative embodiment, the AP and PO in powder form suspended liquid mixed phase.

根据本发明的生产方法一种有趣的实施方式,与至少一种活性成分(AP)结合的PO聚合物的微颗粒分散于基本上水性的液体介质中,所述介质优选含有分散措施M,,并且冻干所得的分散体。 According to the production method of the present invention is an interesting embodiment, in combination with at least one active principle (AP) PO polymer microparticles dispersed in a substantially aqueous liquid medium, preferably the medium containing a dispersion measure M ,, and the resulting dispersion was lyophilized.

因此,所得冻干物有助于制备基于微颗粒的液体制剂(下面所述的本发明的第三方面),因为这种冻干物在用于制备液体制剂的重构液体中分散迅速。 Thus, based on the resultant lyophilisates helpful for preparing the liquid formulation of microparticles (a third aspect of the present invention is described below), since such a lyophilisate for reconstitution liquid disperse rapidly in the liquid preparation.

优选地,所述分散措施Mi选自以下组: Preferably, the dispersion measure Mi is selected from the group:

i. 和PO聚合物的可电离基团极性相反的多价离子,并且其包含在连续的水性相中; . I ionizable polar group and PO polymers may be opposite to polyvalent ions, and which contains in the continuous aqueous phase;

ii. 加入到待雾化的PO悬浮液/溶液中并且因此包含在雾化后的PO/AP微颗粒中的至少一种亲水性化合物(优选能用于可注射的制剂); . Ii was added to the suspension to be atomized PO / contained in the solution and thus atomized PO / AP microparticles of at least one hydrophilic compound (preferably capable of formulation for an injectable);

iii. 用至少一种亲水性化合物(优选能用于可注射的制剂)包 iii. with at least one hydrophilic compound (preferably injectable formulations can be used) packet

至少一层膜的至少一种微颗粒; At least one of the at least one membrane microparticles;

iv. 改变pH; . Iv changing the pH;

V. 措施(i)-(iv)中至少两种的结合; V. steps (i) - at least two of (iv) in the binding;

其中措施(i)特别优选。 Special measures which (i) is preferred. 关于措施M, (i)-(iv)实施方式的其他细节,可参考下面关于分散措施M2的描述。 On measures M, (i) - other details (iv) embodiment embodiment, the dispersion may be reference to the following description of the measures M2.

同样地,关于分散的液体介质,它可以含有和下面所定义的重构液体的添加剂相同的添加剂。 Similarly, with respect to the dispersion liquid medium, which may contain the same additives and the reconstitution liquid additive as defined below.

本发明的第三方面:基于微颗粒的液体制剂,所述微颗粒通过雾化含有AP A third aspect of the present invention: based liquid formulation of microparticles, said microparticles containing AP by atomization

的PO的溶液或胶体悬浮液获得 PO is a solution or colloidal suspension obtained

根据本发明的制剂可以是用于AP延长释放的液体药物制剂,其含有基于和至少一种AP相结合的PO微颗粒的低粘度的胶体悬浮液,这些微颗粒是前面定义的那些或者由前面定义的方法本身和下面的实施例中的方法本身所获得的那匙。 The formulation of the invention may be used to extend the AP release liquid pharmaceutical formulation comprising microparticles based PO AP and at least one combination of a colloidal suspension of low viscosity, the microparticles are those previously defined, or from the front that spoon itself and the methods defined in the following examples per se obtained.

这种制剂的优点是可以通过胃肠外注射并且在注射条件下是液体。 The advantage of this preparation is by parenteral injection liquid and is injected under the condition. 根据本发明,术语"低粘度"或"极低粘度"优选对应于2(TC时的动力学粘度低于或等于lOOOmPa.s。优选地,该制剂的动力学粘度,以2(TC时1000 s"的剪切梯度测定,优选小于或等于500 mPa.s,特别优选2-200 mPa.s,例如1.0-100 mPa.s或1.0-50 mPa.s。 动力学粘度的测定 According to the present invention, the term "low viscosity" or "low viscosity" preferably corresponds to 2 (kinetic viscosity at TC of less than or equal lOOOmPa.s. Preferably, the dynamic viscosity of the formulation to 2 (TC 1000 shear gradient measurement s ", preferably less than or equal to 500 mPa.s, particularly preferably 2-200 mPa.s, measured e.g. 1.0-100 mPa.s or dynamic viscosity of 1.0-50 mPa.s.

动力学粘度的对照测量可以在例如2(TC时使用配备锥/板式几何学(4 cm, 2°) 的AR1000流变仪(TA仪器)上完成。粘度v在10 s"的剪切梯度测定。 Dynamic viscosity control measurements may be performed on, for example, 2 (with the use of a cone / plate geometry when TC (4 cm, 2 °) a AR1000 rheometer (TA Instruments). The viscosity v is 10 s "measured shear gradient .

低粘度可以使本发明的制剂容易以胃肠外方式注射,尤其是通过粘膜、皮下、肌内、皮内、腹腔或脑内途径或注射到肿瘤中。 Low viscosity formulations of the invention can be easily injected parenterally, in particular by a mucosal, subcutaneous, intramuscular, intradermal, intraperitoneal or intracerebral route or into a tumor. 根据本发明的制剂也尤其可以通过口腔、鼻腔、肺部、阴道或眼部途径给药。 In particular it may also be nasal, pulmonary, vaginal, or ocular routes of administration through the oral cavity, the formulations according to the present invention.

本发明的液体制剂的低粘度可同时存在于对应于室温的注射温度(例如4-3(TC)和生理温度。 Low viscosity liquid formulation of the present invention may exist in the room corresponding to the injection temperature (e.g. 4-3 (TC) and the physiological temperature simultaneously.

34而且,通过两性分子PO(例如聚氨基酸)雾化形成的干燥的微颗粒可以容易地再分散以形成低粘度的微颗粒悬浮液或溶液。 34 Further, the dried microparticles formed by amphiphilic molecules PO (such as poly amino acids) can be readily atomized to form fine particles redispersed suspension or solution of low viscosity. 分散措施N^ Dispersion measures N ^

特别关于在重构液体中分散或者悬浮雾化的PO/AP微颗粒以制备制剂,根据本发明提供了所述制剂包含措施M2,用于分散PO/AP微颗粒。 In particular regard to the reconstitution liquid dispersion or suspension atomized PO / AP microparticles to prepare a formulation, the formulation is provided comprising M2 measures according to the invention, for dispersing PO / AP microparticles.

这种分散措施M2可以在性质上和用于形成所述制剂的悬浮液的连续相(即重 This dispersion continuous phase may be used to measure M2 formed on the formulation and the nature of the suspension (i.e., weight

构液体)不同。 Structure of liquid) different.

因此,根据本发明可以尤其展望四种可能性: Thus, according to the present invention may in particular be four possibilities Outlook:

1. 悬浮液的连续相基本上是水性的。 1. The continuous phase of the suspension is substantially aqueous.

2. 悬浮液的连续相基本上是有机的和水互溶的相。 2. The suspension of the continuous phase is substantially immiscible organic and aqueous phases.

3. 悬浮液的连续相基本上是有机的和水不互溶的相。 3. The continuous phase of the suspension and the substantially water-immiscible organic phase.

4. 悬浮液的连续相基本上是有机的和水互溶或不互溶的相。 4. The continuous phase of the suspension is substantially water-miscible and organic or immiscible phase. 术语"基本上水性的连续相"的意思是,例如含有至少60%重量的水的液体。 The term "substantially aqueous continuous phase" is meant, for example water containing at least 60% by weight of the liquid.

术语"基本上有机的连续相"的意思是,例如含有至少60%重量的有机相的液体。 The term "substantially continuous organic phase" means, for example, a liquid organic phase containing at least 60% by weight.

i • 暴淳统的逢续报綦.本i;是杰性的 i • storm Chun Qi system of every newspaper continued this i;. Jay is of

可以展望几种类型的分散措施M2,其可选地相互结合,即所述分散措施M2 It is envisaged that several types of dispersion measures M2, optionally bonded to each other, i.e. the dispersion measures M2

例如选自以下组: For example selected from the following group:

i. 和po聚合物的可电离基团极性相反的多价离子,其包含在 Polar ionizable group multivalent ions of the opposite i. Po and a polymer, which is contained in

连续的水性相中; The continuous aqueous phase;

ii. 加入到待雾化的PO悬浮液/溶液中并且因此包含在雾化后的PO/AP微颗粒中的至少一种亲水性化合物(优选能用于可注射的制剂); . Ii was added to the suspension to be atomized PO / contained in the solution and thus atomized PO / AP microparticles of at least one hydrophilic compound (preferably capable of formulation for an injectable);

iii. 用至少一种亲水性化合物(优选能用于可注射的制剂)包至少一层膜的至少一种微颗粒; . Iii with at least one hydrophilic compound (preferably injectable formulations can be used) coated microparticles of at least one of the at least one membrane;

iv. 改变pH; . Iv changing the pH;

V. 措施(i)-(iv)中至少两种的结合; 其中措施(i)特别优选。 V. steps (i) - (iv) in connection with at least two; wherein special measures (i) is preferred. (i)基于多价离子的分散措施当PO带有可电离基团IG时,所述分散措施M2可以含有多价离子,其极性和聚合物PO的可电离基团IG(至少部分电离的)相反,重构悬浮液/形成悬浮液的溶液时将其加入到水性连续相中。 (I) a dispersion measure based on multivalent ions PO when the IG having ionizable groups, M2 measures the dispersion may contain multivalent ions, and the polarity of the polymer PO ionizable group IG (at least partially ionized when) in contrast, the reconstituted suspension / solution of the suspension formed which was added to the aqueous continuous phase.

根据本发明,术语"多价离子"指例如二价离子、三价离子、四价离子以及这些离子的混合物。 According to the present invention, the term "multivalent ions" means, for example divalent ions, trivalent ions, tetravalent ions and mixtures of these ions.

当PO具有阴离子基团IG时,所述多价离子是多价阳离子,优选二价阳离子,并且特别优选自包括Mg"、 Ca2+、 Zn2+、 Fe2+、 012+及其混合物的组,和/或三价离子,特别优选自包括A产、F^+或其混合物的组。 When PO having an anionic group the IG, the multivalent ion is a multivalent cation, preferably a divalent cation, and particularly preferably from the group consisting of Mg ", Ca2 +, Zn2 +, Fe2 +, 012+ and mixtures thereof, and / or tris divalent ions, preferably selected from the group consisting of a particularly yield, F ^ + or mixtures thereof.

除多价离子外,根据本发明的制剂可以包括单价离子(例如阳离子),它们在纳米颗粒聚集成微颗粒的过程中可以起作用。 In addition to the polyvalent ions, the formulation according to the present invention may comprise monovalent ion (e.g. cation), are aggregated into fine particles during the nanoparticles may function.

这些多价离子优选以水性(有机或矿物)盐溶液的形式,例如多价离子的硫酸盐、氯化物、乙酸盐、葡糖酸盐或谷氨酸盐(或其它阴离子氨基酸)的溶液加入到本发明的制剂中。 These multivalent ions preferably in the form of an aqueous salt solution (organic or mineral), polyvalent ions such as sulfate, chloride, acetate, gluconate or glutamate (amino acids or other anionic) was added to the formulation of the present invention.

这种基于多价离子的分散措施M2优选适用于两性分子PO(例如一种(共)聚氨基酸)是相对亲水的情形。 This dispersion measure based on multivalent ions M2 is preferably suitable amphiphilic molecules PO (eg, one (co) poly-amino acids) of the case are relatively hydrophilic.

(ii)和(iii)基于亲水性化合物或含有至少一种基于亲水性化合物的包膜的分散措施M2 (Ii) and (iii) based on the hydrophilic compound or at least one dispersing action based on an envelope containing a hydrophilic compound M2

该分散措施M2可以主要包括: M2 measure of the dispersion may include:

-加入到待雾化的PO悬浮液/溶液中并且因此存在于雾化的PO/AP微颗粒中的至少一种亲水性化合物(优选其能用于可注射的制剂); - was added to the suspension to be atomized PO / present in the solution and thus atomized PO / microparticles of the AP at least one hydrophilic compound (which preferably can be used in injectable formulations);

-或者,用至少一种亲水性化合物膜(优选其能用于可注射的制剂)对微颗粒包上至少一层膜。 - or with at least one hydrophilic compound film (which preferably can be used for injectable formulation) at least one layer of film on the pack microparticles.

这种分散措施M2可以按照几种方法结合到制剂中。 This dispersion may be incorporated into the M2 measures the formulation according to several methods.

根据第一种方法,至少一种亲水性化合物加入到待雾化的PO悬浮液/溶液中,这种亲水性化合物选自可用于可注射制品的那些,特别优选自包括以下的 According to a first method, at least one hydrophilic compound to the PO suspension / solution to be atomized, such hydrophilic compound selected from those used in the preparation of injectables, particularly preferably from the group consisting of the following

组: group:

— 氨基酸类(例如赖氨酸、精氨酸、甘氨酸); - amino acids (e.g. lysine, arginine, glycine);

4 聚二醇类,优选聚乙二醇类; 4 polyalkylene glycols, preferably polyethylene glycols;

~> 共聚二醇类,优选(Poloxamer、 Pluronic或Lutrol型的)乙二醇/丙二醇 ~> Copolymerizable glycols, preferably (Poloxamer, Pluronic or Lutrol type) of ethylene glycol / propylene glycol

共聚物; Copolymers;

36— 纤维素聚合物类及其衍生物,优选羧基垸基纤维素(例如羧甲基纤维素)或烷基纤维素(例如甲基纤维素); 36- cellulose polymers and derivatives thereof, carboxy alkyl with preferably a cellulose (e.g., carboxymethylcellulose), or an alkyl cellulose (e.g. methylcellulose);

4 氢化或非氢化的糖类,例如海藻糖、山梨醇、甘露醇或蔗糖; 4 hydrogenated or non-hydrogenated saccharides, such as trehalose, sorbitol, mannitol, or sucrose;

— 多元醇类,如丙二醇或甘油; - polyhydric alcohols such as propylene glycol or glycerol;

— 明胶类,优选水解明胶; - gelatin preferably hydrolyzed gelatin;

— 含氮(共)聚合物类,优选自包括聚丙烯酰胺、聚-N-乙烯酰胺、聚乙烯吡咯烷酮(PVP)和聚-N-乙烯内酰胺的组; - a nitrogen-containing (co) polymers, preferably from the group comprising polyacrylamide, poly -N- vinyl amide, polyvinyl pyrrolidone (PVP) and poly-vinyl lactam -N- groups;

— 聚(乙烯醇)类(PVA); - poly (vinyl alcohol) (the PVA);

— 聚(谷氨酸钠); - poly (sodium glutamate);

— 及其混合物; - and mixtures thereof;

所述亲水性包膜化合物优选含有至少一种亲水性聚合物。 Said envelope hydrophilic compound preferably contains at least one hydrophilic polymer.

根据第二种方法,所述微颗粒包有用前面定义的至少一种亲水性化合物的 According to the second method, the microparticles includes at least one hydrophilic compound is useful to previously defined

至少一层。 At least one layer.

在该第二种方法中,所述亲水性化合物优选含有至少一种选自前面定义的亲水性化合物的亲水性聚合物。 In this second method, it said hydrophilic compound preferably contains at least one hydrophilic compound selected from the previously defined hydrophilic polymer.

基于至少一种亲水性化合物的分散措施M2 (ii)被证明为特别适合疏水性足够高的两性分子PO,例如对于由至少一种聚氨基酸构成的PO,其HG基团的水平大于或等于10mol%。 Decentralized measures at least one hydrophilic compound M2 (ii) is found to be particularly suitable for hydrophobic amphipathic molecules PO sufficiently high, for example, PO consisting of at least one polyamino acid, which HG level greater than or equal groups 10mol%.

用这种亲水的和生物相容的化合物对微颗粒的包膜[措施M2 (iii)]可以通过例如两步连续的雾化进行,其中第二步的雾化在和微颗粒不互溶的溶剂中在颗粒的悬浮液上进行,所述溶剂有助于这些颗粒的分散。 Coated microparticles [measures M2 (iii)] may be used such hydrophilic and biocompatible compound, for example, by atomizing a continuous two steps, wherein in the second step and atomization microparticles immiscible in the solvent in the suspension of particles, the solvent help disperse the particles.

通过对微颗粒进行亲水性包膜的这种分散措施(iii)可以使微颗粒的悬浮液保持可靠和稳定至少达几天,这使其易于操作。 By this measure the dispersion (iii) of the microparticles may be coated hydrophilic microparticles of the suspension remains stable for at least up to a few days and reliable, making it easy to operate.

优选地,将用于分散基于亲水性化合物的微颗粒的措施(ii)或(m)和在制剂重 Preferably, based on measures for dispersing microparticles of a hydrophilic compound (ii) or (m) and the weight of the formulation

构期间用于分散存在于水性相中二价离子的措施(i)相结合可以加速分散。 During configuration for dispersing the aqueous phase in the presence of divalent ions steps (i) combining the dispersion can be accelerated. 根据另一种具体实施方式,该水性连续相或者亲水性包膜也可以含有至少 According to another particular embodiment, the aqueous continuous phase or the hydrophilic envelope may also contain at least

一种可注射的表面活性剂,例如Tween⑧80、卵磷脂、磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰丝氨酸或者聚氧丙烯/聚氧乙烯共聚物(商品名:Poloxamer、 Pluronic、 Lu加l)。 Surfactants an injectable, e.g. Tween⑧80, lecithin, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine or polyoxypropylene / polyoxyethylene copolymer (trade name: Poloxamer, Pluronic, Lu added l).

(iv)通过改变pH的分散措施M2根据本发明可以展望的另一种分散措施是通过改变pH的分散措施,优选在雾化之前。 (Iv) by varying the dispersion pH M2 measures according to another measure dispersion of the present invention may be envisioned by dispersing measure change in pH, preferably before atomization. 己证明这种类型的措施尤其适合于其中两性分子PO(例如聚氨基酸) 是带有可电离官能团并且高度亲水的化合物的情形,也就是说,其含有例如低 This type of measure has proved particularly suitable in which the amphiphilic molecules PO (e.g. poly amino acid) compound is the case with ionizable functional groups and a highly hydrophilic, i.e., containing, for example low

于10moP/。 In 10moP /. 的疏水性基团HG。 Hydrophobic group HG.

事实上,采用这种措施M2 (iv)能够通过将pH值调节至使PO(例如聚氨基酸) 的可电离官能团不发生电离(例如当PO是聚Glu或聚Asp型的亲水性聚氨基酸时酸化)的值来提高两性分子PO的疏水性。 In fact, using this measure M2 (iv) can be prepared by adjusting the pH to make PO (such as poly amino acids) can not ionize ionizable functional groups (when PO is a hydrophilic poly Glu or Asp poly amino acid type, for example polyethylene acidification) to increase the value of amphiphilic PO hydrophobic.

优选地,这种通过在雾化前改变pH的分散措施M2 (iv)应用于PO悬浮液/ 溶液后立即雾化。 Preferably, this suspension is applied by altering the PO pH of the dispersion measure M2 (iv) atomizing before / immediately after the atomized solution.

这种通过在雾化前改变pH的分散措施M2 (iv)可以和以下措施联合或者不联合使用: Such a joint can not be used in combination or by dispersing the following measures and measures to change the pH before atomization M2 (iv):

,在雾化后的重构步骤中使用的基于多价离子的分散措施M^i);或者,基于亲水性化合物的分散措施M2(ii)或(iii),优选通过用至少一种亲水性聚合物包膜M^iii)微颗粒。 , Based on the dispersion measures multivalent ions M ^ i) used in the reconstruction step atomized; alternatively, a dispersion measure based on a hydrophilic compound M2 (ii), or (III), preferably by treatment with at least one affinity The aqueous polymer envelope M ^ iii) microparticles.

2. 悬.浮液.的逢续报棊.本J;是有机的.租杰.互.澄放报 . 2. The suspension of the present Qi every followup J;. Mutual organic rent Jie Cheng discharge reported..

分散措施由和水互溶的有机相组成。 Measures of dispersion and a water-miscible organic phase.

该相因此可以例如,含有乙醇、N-甲基吡咯垸酮(NMP)、 二甲基亚砜(DMSO)、异丙醇、甘油或三縮四乙二醇(四氢糠基醇聚乙二醇醚)。 The phase can thus be, for example, containing ethanol, N- methyl pyrrol embankment one (NMP), dimethyl sulfoxide (DMSO), isopropanol, glycerol or glycofurol (tetrahydrofurfuryl alcohol polyethylene glycol ether).

3. 暴浮痕的逢续报蓋.本丄.是有机的报杰丕瓦澄的报根据一个优选的实施方式,分散措施含有一种存在于和水不互溶的有机连 3. Floating marks every storm continued report cover. Shang present. Jay organic pi watt clear message packets in accordance with a preferred embodiment, the dispersion contains a measure and present in a water-immiscible organic even

续相中的亲脂性液体,其熔点优选低于或等于15"C。 Lipophilic liquid continuous phase having a melting point preferably less than or equal to 15 "C.

优选地,该亲脂性液体含有至少一种饱和脂肪酸的甘油三酯的混合物或者至少一种植物油或至少一种脂质或至少一种脂质衍生物或至少一种脂肪酸或至少一种脂肪酸衍生物。 Preferably, the lipophilic liquid comprises a mixture of at least one saturated fatty acid triglyceride or at least one vegetable oil or at least one lipid or lipid derivative, or at least one of the at least one fatty acid or at least one fatty acid derivative .

更优选地,该亲脂性液体含有: More preferably, the lipophilic liquid comprises:

•源自椰子油的饱和C8-C1Q脂肪酸的甘油三酯的混合物,例如由Sasol以"Miglyol812"出售的那种; • a mixture of triglycerides derived from coconut oil fatty acids, saturated C8-C1Q, for example by Sasol in "Miglyol812" sold under that;

,至少一种植物油,优选大豆油、棕榈油、亚麻籽油、棉籽油、芝麻油、葵花籽油或花生油; At least one vegetable oil, preferably soybean oil, palm oil, linseed oil, cottonseed oil, sesame oil, sunflower oil, or peanut oil;

,至少一种脂质,优选脂质卵磷脂、合成或天然的维生素E;•至少一种脂质衍生物,优选花生四烯酰磷脂酰胆碱和硬脂酰磷脂酰胆碱; *至少一种脂肪酸,优选油酸、肉豆蔻酸、棕榈酸、硬脂酸及其盐类; At least one lipid, preferably phosphatidylcholine lipids, synthetic or natural vitamin E; • at least one lipid derivative, preferably arachidonic stearoyl phosphatidyl choline and phosphatidyl choline; * at least one fatty acids, preferably oleic acid, myristic acid, palmitic acid, stearic acid and salts thereof;

*至少一种脂肪酸衍生物,优选单-、双-或三-甘油酯衍生物、油酸乙酯、乳酸月桂酯、硬脂酸甘油酯、棕榈酸脱水山梨聚醇酯、硬脂酸脱水山梨醇酯、脱水山梨醇一油酸酯或聚山梨醇酯; * At least one fatty acid derivative, preferably mono -, bis - or tri - glyceride derivatives, ethyl oleate, lauryl lactate, glyceryl stearate, palmitic acid, polyethylene sorbitan esters, sorbitan stearate alcohol esters, sorbitan monooleate or polysorbate;

*和任何它们的混合物; * And any mixtures thereof;

其条件是当某些上面列出的产品如果在例如低于或等于i5X:时单独使用时 With the proviso that when some of the products listed above, for example, if less than or equal i5X: when used alone

不是液体形式,那么这些产品可以和其它产品混合,以使其在例如低于或等于15"C时是液体形式。 Not in liquid form, then these products can be mixed with other products, for example, so as to equal to or less than 15 "C in liquid form.

特别优选脂肪酸甘油三酯的衍生物,尤其是源自椰子油的饱和C8-do脂肪酸的甘油三酯的混合物,例如由Sasol以"Migiyol 812"出售的那种。 Particularly preferred derivatives of fatty acid triglycerides, in particular triglycerides derived from coconut oil fatty acids, saturated C8-do mixture, such as that by Sasol to "Migiyol 812" sold. 其它含有脂肪酸长链的甘油三酯尤其存在于植物油中,例如大豆油、棕榈油、亚麻籽油、棉籽油、芝麻油、葵花籽油或花生油。 Other long-chain fatty acid containing triglycerides, in particular in vegetable oils, such as soybean oil, palm oil, linseed oil, cottonseed oil, sesame oil, sunflower oil or peanut oil.

雾化的PO/AP粉末易于在这种和水不互溶的有机相中分散以得到稳定的微颗粒悬浮液,其可以保持稳定达数天之久,因此容易操作。 Atomized PO / AP in this powder and the easily water-immiscible organic phase to obtain a stable dispersion of microparticles suspension, which may be stable for several days, it is easy to operate. 这种分散尤其迅速,其不用加入另外的或其它分散措施,即使可以展望这种可能性。 This particular the rapid dispersion, which was used without additional or other measures to disperse, even though this possibility may be envisaged.

4- 悬.淫液節.逢续相.蒸.本丄.悬有.机.的招.杰s澄的或煮.和水丕.瓦澄 4- suspended. Yinye section. Every continuous phase. Evaporated. Shang present. Hanging there. Dryers. Strokes. S Jie Cheng broiled. Pi and water. Cheng W

.的报 . The report

根据一个优选的、和构成制剂的悬浮液/溶液的连续相的性质有关的第二和第三种可能性相兼容的方法的可供选择的方式,这种分散措施含有用至少一种膜形成包膜化合物(优选其能用于可注射的制剂)对微颗粒进行包膜。 Alternatively the second and third possibility manner according to the nature of a preferred continuous phase, comprising a formulation, and suspension / solution compatible related method, which measures the dispersion comprising at least one film-forming envelope compound (which preferably can be used for injectable preparations) to be coated microparticles.

优选地,这种膜形成包膜化合物含有至少一种疏水性聚合物,选自包括以下物质的组:聚交酯、聚羟基乙酸、聚(交酯-共-乙交酯)、聚原酸酷、聚酸酐、 聚(羟基丁酸)、聚己酸内酯、聚(碳酸烷基酯)、水不溶性PO聚合物、它们的衍生物和混合物。 Preferably, such compound film envelope containing at least one hydrophobic polymer selected from the group comprising the following substances: polylactide, polyglycolic acid, poly (lactide - co - glycolide), poly ortho cool, polyanhydrides, poly (hydroxybutyric acid), polycaprolactone, poly (alkyl carbonate), water-insoluble PO polymers, derivatives and mixtures thereof.

根据一种可供选择的形式,这种膜形成包膜化合物是脂质性质的,其烙点优选高于或等于15'C,并且含有至少一种脂质或至少一种脂质衍生物,或至少一种植物油,或至少一种脂肪酸或至少一种脂肪酸衍生物或至少一种饱和脂肪酸的甘油三酯的混合物或这些产品的混合物。 According to an alternative form of this compound is a lipid envelope film forming properties, which point is preferably higher than or equal to branded 15'C, and containing at least one of the at least one lipid or lipid derivative, or at least one vegetable oil, or at least one fatty acid or at least one fatty acid derivative, or a mixture of at least one saturated fatty acid triglyceride, or of these products.

根据另一个优选的实施方式,这种有机的连续相,例如疏水性的,或者疏水性包膜可以含有至少一种可注射的表面活性剂,例如Tween⑧80、卵磷脂、磷脂酰乙醇胺、磷脂酰丝氨酸或者聚氧丙烯/聚氧乙烯共聚物(商品名: Poloxamer、 Pluronic、 Lutrol)。 According to another preferred embodiment, the organic continuous phase, such as hydrophobic, or hydrophobic envelope may contain at least one surfactant injectable, e.g. Tween⑧80, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine or a polyoxypropylene / polyoxyethylene copolymer (trade name: Poloxamer, Pluronic, Lutrol).

鹏浪剂/.稳窓.剂 Peng waves agents / stabilizing Chuang. Agent

也可以是优选的,以利于在水性相中分散的性质和/或进一步提高水性悬浮液的稳定性,可注射的制剂可以含有其他添加物, 一方面是下面所指的"赋形剂/ 稳定剂",另一方面是传统的赋形剂。 May also be preferred to facilitate dispersion in the aqueous phase, the nature and / or further enhance the stability of the aqueous suspension, injectable formulations may contain other additives, one is referred to below "excipient / stabilizer agent ", on the other hand is the traditional excipients.

赋形剂/稳定剂可以选自包括以下的组: Excipients / stabilizers may be selected from the group consisting of:

— 至少一种聚合物PO的纳米颗粒,PO是水溶性的可生物降解的两性 - at least one polymer nanoparticles of PO, PO is a water-soluble biodegradable amphoteric

分子共聚物,带有疏水性基团(HG)和至少部分电离的可电离亲水性基团(IG),其在pH = 7.0,等渗条件下自发地形成一种纳米颗粒的胶体悬浮液; 4 氨基酸; Ionizable hydrophilic group copolymer molecule, having a hydrophobic group (HG) and the at least partially ionized (the IG), which in the pH = 7.0, form a colloidal suspension of nano particles spontaneously under isotonic conditions ; 4 amino acids;

— 聚二醇类,优选聚乙二醇; - polyalkylene glycols, preferably polyethylene glycol;

— 共聚二醇类,优选乙二醇/丙二醇共聚物(Poloxamer、 Pluronic或Lutrol型的); - co glycols, preferably ethylene / propylene copolymer (Poloxamer, Pluronic or Lutrol type);

— 纤维素聚合物及其衍生物,优选羧烷基纤维素(例如羧甲基纤维素) 或垸基纤维素(例如甲基纤维素); - cellulosic polymers and derivatives thereof, preferably carboxyalkyl cellulose (e.g., carboxymethylcellulose), or alkyl with cellulose (e.g. methylcellulose);

— 山梨聚糖和一种或多种脂肪酸的酯类,优选吐温(或聚山梨醇酯)类的聚氧烯烃(例如聚氧乙烯)二醇和至少一种酸(例如油酸)的酯类; - esters of sorbitan and one or more fatty acids, preferably Tween (Polysorbate) like polyoxyalkylene (e.g. polyoxyethylene) glycol and at least one acid (e.g., oleic acid) esters ;

— 基于磷脂和聚烯烃二醇的表面活性剂,优选聚乙二醇; - phospholipid-based surfactants and polyolefin diols, preferably polyethylene glycol;

— 氢化或非氢化的糖类,例如海藻糖、山梨醇、甘露醇或蔗糖; - hydrogenated or non-hydrogenated saccharides, such as trehalose, sorbitol, mannitol, or sucrose;

— 多元醇,如丙二醇或甘油; 4 朋胶,优选水解明胶; - polyhydric alcohols such as propylene glycol or glycerol; 4 Four plastic, preferably hydrolyzed gelatin;

~> 含氮(共)聚物,优选自含有聚丙烯酰胺、聚-N-乙烯酰胺、聚乙烯吡 ~> A nitrogen-containing (co) polymer, preferably contain from polyacrylamide, poly -N- vinyl amide, polyvinyl pyrazole

咯烷酮(P VP)和聚-N-乙烯内酰胺的组; Pyrrolidone (P VP) and vinyl lactam poly -N- group;

— 聚(乙烯醇)类(PVA); - poly (vinyl alcohol) (the PVA);

及其混合物。 And mixtures thereof.

根据本发明的优选的赋形剂(稳定剂)中的一种由至少一种聚合物PO的纳米颗粒构成,所述PO和构成所述微颗粒的PO相同或不同(优选相同)。 According to a preferred of the present invention, excipients (stabilizers) is composed of at least one polymer PO nanoparticles, the same or different PO and PO constituting said microparticles (preferably the same).

制剂中所用赋形剂/稳定剂的量优选0.01-10%重量,特别优选0.1-5%重量。 The formulation excipients / stabilizers is preferably 0.01 to 10% by weight, particularly preferably 0.1 to 5% by weight.

40至于含有纳米颗粒的稳定剂,它们优选以1.5-3.5%重量的比例用于所述制 40 As a stabilizer of the nanoparticles, they are preferably in a ratio of 1.5 to 3.5% by weight for the system

齐U,例如2.0-3.0% (- 2.5%)重量。 Qi U, e.g. 2.0-3.0% (- 2.5%) by weight.

重构液体 Reconstruction of liquid

除了前面描述的分散措施,根据本发明的制剂的制备中所用的重构液体可 In addition to the measures described above dispersion, prepared according to the reconstituted liquid formulation according to the invention may be used in the

以含有,例如: Containing, for example:

-至少一种缓冲液或至少一种可注射盐(磷酸盐缓冲液、拧檬酸盐缓冲 - at least one buffer or at least one injectable salt (phosphate buffer, citric acid buffer

液、氯化钠),浓度为例如0.001 M-0.1 M,优选0.005 M-0.02 M,这种缓冲 Solution, sodium chloride) concentration, for example 0.001 M-0.1 M, preferably 0.005 M-0.02 M, such a buffer

液或这种可注射盐能够调节溶液的pH;和 Such injectable solutions or salts capable of adjusting the pH of the solution; and

-至少一种可注射的表面活性剂,优选聚山梨醇酯型的,例如 - at least one surfactant injectable, preferably polysorbate type, e.g.

Tween® 20或Tween® 80 ,或者是Poloxamer类的,例如Lutrol® F3 8 、 Lutrol⑧F68或Lutro1⑧F127,浓度为例如0.01%-2%,优选0.05-0.5%。 Tween® 20 or Tween® 80, class or Poloxamer, e.g. Lutrol® F3 8, Lutrol⑧F68 or Lutro1⑧F127, for example, a concentration of 0.01% to 2%, preferably 0.05 to 0.5%.

重构液体还可以包括增加密度的试剂,例如糖类,尤其是蔗糖、D-甘露醇或海藻糖,浓度为0.1%-10%,优选0.5-5%。 The reconstitution liquid may also include an agent to increase the density, for example, sugars, in particular sucrose, D- mannitol, or trehalose, at a concentration of 0.1-10%, preferably 0.5-5%. 这种重构溶液还可以含有可注射的增粘聚合物,选自包括多糖、合成聚合物(例如羧甲基纤维素钠)、聚乙烯醇、聚乙烯吡咯烷酮、聚二醇(如聚乙二醇)及其混合物的组。 This reconstituted solution may further contain a tackifier injectable polymer selected from the group comprising polysaccharides, synthetic polymers (e.g. sodium carboxymethyl cellulose), polyvinyl alcohol, polyvinylpyrrolidone, polyethylene glycols (e.g., polyethylene alcohols) and mixtures of.

两性分子PO(例如聚氨基酸)干燥微颗粒的使用和即用的(ad hoc)重构措施的使用因此能够满足双重目标,能够获得既稳定又易于分散的药物制剂,以使其能够胃肠外注射。 Amphiphilic PO (such as poly amino acids) that use the dried microparticles and (ad hoc) that is reconstituted measures can be used to meet the dual objectives, to obtain stable and readily dispersible pharmaceutical formulations, so that it can be parenterally injection.

除了前面所列的用于改善分散和稳定性的赋形剂/稳定剂以外,其它可以加入到待雾化的悬浮液/溶液或根据本发明的制剂中的常用赋形剂是,例如是本领域技术人员所知的抗微生物剂、缓冲剂、抗氧化剂和渗透压调节剂。 In addition to the previously listed excipients for improving / stabilizer dispersion and stability, it can be further added to the suspension / solution or excipients commonly used in the formulation according to the present invention is to be atomized, for example, are skill in the art antimicrobial agents, buffers, and antioxidants, osmolality adjusting agent. 可以参考例如Injectable Drug Development—书,PK Gupta等,Interpharm Press, Denver, Colorado, 1999。 Injectable Drug Development- reference may be, for example, books, PK Gupta, etc., Interpharm Press, Denver, Colorado, 1999.

常用赋形剂可以加入到水性分散相中或者在雾化前加入到溶液/悬浮液中。 Common excipients may be added to the aqueous or dispersed phase is added to the solution / suspension prior to the atomization. 根据本发明的液体制剂的应用 The application of liquid formulations of the present invention.

虽然根据本发明的制剂优选是药物,但这并不排除化妆品、健康食品或植物保护制剂,其包括至少一种前面定义的PO和至少一种AP。 Although the preferred formulations of the present invention is a pharmaceutical, this does not exclude cosmetic, a health food or a plant protection formulation comprising at least one PO and at least one AP defined above.

本发明第四方面:用于重构根据本发明制剂的药盒 A fourth aspect of the present invention: A kit for reconstitution of the formulation according to the invention

PO/AP微颗粒和基本上水性的重构液体、基本上有机的和水互溶的重构液体和基本上有机的和水不互溶的重构液体已在前面定义。 PO / AP microparticles and substantially aqueous reconstitution liquid, substantially organic and reconstitution liquid and substantially water-miscible and water-immiscible organic liquids have been reconstructed defined previously. 本发明第五方面:重构方法,尤其是重构根据本发明制剂的方法PO/AP微颗粒和基本上水性的重构液体、基本上有机的和水互溶的重构液体和基本上有机的和水不互溶的重构液体已在前面定义。 The fifth aspect of the present invention: reconstruction methods, in particular reconstituted preparation method according to the present invention PO / AP microparticles and substantially aqueous reconstitution liquid, substantially water immiscible organic liquid and reconstituted substantially organic and a water-immiscible liquid have been defined previously reconstructed.

根据本发明,可以展望使用过滤除菌的方式,在孔径例如等于0.2 pm的滤膜上过滤纳米颗粒悬浮液,生成根据本发明的制剂的微米尺度的颗粒。 According to the present invention, use may be envisaged filter sterilized manner, for example equal to the pore size of the filter on the filter nanoparticle suspension of 0.2 pm, the particles generated according to microscale formulation of the present invention. 因此根据前面描述的方法在无菌条件下的聚集可以将制剂直接注射给病人。 Thus gathered under aseptic conditions can be injected directly formulation according to the method described above to the patient.

本发明第六方面:固体药物制剂 A sixth aspect of the present invention is: a solid pharmaceutical formulation

这种用于AP延长释放的制剂含有用于吸入或肺部给药的基于己在前面定义 Such extended release formulations for containing AP for inhalation or pulmonary administration based on already defined previously

的PO/AP微颗粒的干燥粉末形式。 The PO / AP dry powder microparticles.

本发明的其它方面 Other aspects of the invention

本发明还涉及衍生自根据本发明的PO微颗粒的固体产品。 The present invention further relates to a solid product derived from PO microparticles of the present invention.

在实践中,这些衍生产品可以尤其由粉末、凝胶、植入物或薄膜组成。 In practice, these derived products can be made in particular powders, gels, implants or films composed. 因此,本发明涉及衍生自根据本发明的制剂的产品,它们与其制备方法无关。 Accordingly, the present invention relates to products derived, are independent of their method of preparation of the formulation according to the present invention product.

本发明还涉及一种治疗方法,其主要在于通过胃肠外、粘膜、皮下、肌内、皮内、腹腔或脑内途径给药或肿瘤内给予根据本发明的制剂。 The present invention further relates to a method of treatment, which mainly by parenteral, subcutaneous, intramuscular, intradermal, intraperitoneal or intracerebral route, or intratumoral administration of the formulation according to the present invention. 也可以尤其通过口服、经鼻、经肺部、经阴道或经眼部途径给予本说明书中描述的制剂。 In particular it may be administered by oral, nasal, pulmonary, vaginal or ocular routes formulation described in this specification is administered.

在本发明的一个具体的可供选择的方式中,该治疗方法在于通过胃肠外、 皮下、肌内、皮内、腹腔或脑内途径注射或注射到肿瘤内给予该制剂,优选以在注射位点形成贮库层的方式给药。 In a specific alternative embodiment of the present invention, wherein the method of treatment by parenteral, subcutaneous, intramuscular, intradermal, intraperitoneal or intracerebral route of injection into the tumor or injection administration of the formulation, preferably in injectable administration site of the reservoir layer is formed.

通过以下实施例将更清楚地理解本发明,且本发明的优点和可供选择的实施方式将更清楚地在实施例中显现,所述实施例描述了由聚氨基酸与疏水性基 Clearly understood by the following examples of the present invention will be more, and advantages of the invention and alternative embodiments will emerge more clearly in the embodiments described by hydrophobic polyamino acids embodiment the group

团接枝形成的PO的合成,以及它们转变为AP的延长释放体系,即转变为本发 Synthesis of PO groups formed by grafting, and their transformation AP extended release system, i.e. converted to present hair

明的制剂(干燥微颗粒的粉末),以及这种体系的稳定性和再分散性。 Clear formulation (dried powder of microparticles), and the stability of such systems and redispersibility. 具体实施方式 Detailed ways

实施例l:两性分子聚合物PO-A的合成 Synthetic amphiphilic polymers of PO-A: Embodiment Example l

接枝了合成来源的cc-生育酚的聚谷氨酸的合成 The synthesis of poly-glutamic acid grafted cc- tocopherol synthetic origin

在8(TC加热15g聚(aL-谷氨酸)(相对于聚氧乙烯标准的分子量约为16900 In 8 (TC heating 15g of poly (aL-glutamic acid) (with respect to a polyoxyethylene standard molecular weight of about 16,900

42Da,其通过聚合NCAGluOMe然后水解得到,如专利申请FR-A-2801226所描述) 直到其溶解在288 ml二甲基甲酰胺(DMF)里。 42Da, which is obtained by polymerization and then hydrolysis NCAGluOMe, as described in patent application FR-A-2801226 described below) until it was dissolved in 288 ml of dimethylformamide (DMF) in. 将溶液冷却至15°C,连续加入预先溶解在8mlDMF里的2.5 gD,La-生育酚(〉98X,购自Fluka⑧)、预先溶解在1 ml DMF里的280 mg 4-二甲基氨基吡啶和预先溶解在6 ml DMF里的1.6 g二异丙基碳二亚胺。 The solution was cooled to 15 ° C, was added continuously in the 8mlDMF previously dissolved in 2.5 gD, La- tocopherol (> 98X, available from Fluka⑧), pre-dissolved in 1 ml DMF in a 280 mg 4- dimethylaminopyridine and pre-dissolved in 6 ml DMF was 1.6 g of diisopropyl carbodiimide. 搅拌3小时后,将反应介质倒入1200 ml含有15n/。 After stirring for 3 hours, the reaction medium is poured into 1200 ml containing 15n /. 的氯化钠和盐酸(pH二2)的水中。 Water, sodium chloride and hydrochloric acid (pH = 2) a. 然后通过过滤回收沉淀的聚合物,并用0.1N盐酸、水和二异丙醚洗涤。 Then the precipitated polymer was recovered by filtration, and washed with 0.1N hydrochloric acid, water and diisopropyl ether. 然后该聚合物在4CTC真空箱中干燥,得到的产率约为90%。 The polymer was then dried in a vacuum oven 4CTC, to give about 90% yield. 用体积排除色谱法测得的相对于聚氧乙烯标准的分子量为15500。 By size exclusion chromatography, the molecular weight measured with respect to a polyoxyethylene standard 15,500. 用质子NMR光谱法评估的生育酚的接枝比例为5.1 mol%。 Tocopherol graft ratio assessed by proton NMR spectroscopy was 5.1 mol%. 将其溶解在水中,并将pH调节(中和羧酸盐)至7±1,获得聚合物在水中的纳米颗粒悬浮液。 Which was dissolved in water, and the pH was adjusted (and carboxylate) to 7 ± 1, to obtain a polymer nanoparticle suspension in water.

实施例2:两性分子聚合物PO-B的合成 Synthesis of amphiphilic polymer PO-B of Example 2:

实施例2由实施例1改变条件进行,目标的接枝率为20%。 Example 2 Example 1 is performed by changing conditions embodiments, the target is 20% grafting.

实施例3:含有IFN-a2b的聚合物PO-A的干燥微米-尺寸的颗粒的制备制备含有20 mg/g聚合物和0.25 mg/g IFN的溶液 Example 3: IFN-a2b dried microns containing polymer of PO-A - Preparation of particle size 20 mg / g of polymer and 0.25 mg / g IFN solution containing

将200 g聚合物PO-A浓度为30 mg/g的溶液加入到500 ml的烧瓶中。 The PO-A polymer concentration of 200 g 30 mg / g was added to a 500 ml flask. 然后向含有聚合物的烧瓶中加入68 g 7jC。 Then the flask containing the polymer was added 68 g 7jC. 将浓縮至2.8mg/g的IFN-a-2b冷冻溶液于25'C解冻1 h,然后将26.8 g解冻的溶液加入到含有聚合物溶液的烧瓶中。 Concentrated to 2.8mg / g of IFN-a-2b solution was freeze-thawed 25'C 1 h, then 26.8 g of the thawed solution was added to the flask containing the polymer solution. 该混合物于室温条件下静置14 h。 The mixture was allowed to stand at room temperature for 14 h.

将该溶液通过0.2pm的除菌滤膜过滤。 The solution was sterilized by filtration through a 0.2pm filter.

聚合物-IFN溶液的雾化 -IFN atomized polymer solution

该溶液在Biichi B290型的喷雾干燥装置上雾化。 The solution is atomized on a Biichi B290-type spray-drying apparatus. 以5 ml/min的速度抽出液体溶液,并通过供氮(7 bar,卯0 1/h)的喷嘴雾化。 At a rate of 5 ml / min was withdrawn liquid, and by supplying nitrogen (7 bar, d 0 1 / h) of the atomizing nozzle. 抽出流速(干空气)为40 m3/h。 Extraction flow rate (dry air) of 40 m3 / h. 进口温度维持在9(TC,这些条件下的出口温度为45。C。 Inlet temperature is maintained at 9 (TC, outlet temperature under these conditions is 45.C.

所得微颗粒的表征: The resulting microparticles characterization:

所得颗粒的尺寸(根据T0试验)为D50 = 5 The resulting particle size (according to test T0) of D50 = 5

将粉末于雾化浓度分散于水中以后,IFN的HPLC检测和雾化前溶液的检测相同,未发现分解形式。 The atomized powder is dispersed in water after concentration, the same is detected by HPLC the solution prior to atomization of IFN and found no decomposition form. 实施例4:含有IFN-a2b和聚山梨醇酯80的聚合物PO-A的干燥微米-尺寸的颗 Example 4: drying microns containing IFN-a2b and polysorbate 80 polymer of PO-A - particle size

粒的制备 Preparation of tablets

PO聚合物和干扰素溶液的制备和实施例3相同,在雾化前将0.9g聚山梨醇酯80加入到溶液中。 Preparation Example 3 and the same PO polymer and interferon solution before the atomization 0.9g of polysorbate 80 was added to the solution.

在聚山梨醇酯存在下雾化聚合物-IFN溶液溶液的雾化条件和实施例3相同。 In the presence of polysorbate same atomizing conditions -IFN polymer solution was atomized and Example 3. 所得微颗粒的表征: The resulting microparticles characterization:

所得颗粒的尺寸(根据T0试验)为D50 = 5pm. The resulting particle size (according to test T0) of D50 = 5pm.

将粉末于雾化浓度分散于水中以后,IFN的HPLC检测和雾化前溶液的检测相同,未发现分解形式。 The atomized powder is dispersed in water after concentration, the same is detected by HPLC the solution prior to atomization of IFN and found no decomposition form.

实施例5:含有IFN-a2b的聚合物PO-A的酸化的干燥微米-尺寸的颗粒的制备 Example 5: IFN-a2b acidified containing polymer PO-A drying micron - particle size prepared

聚合物PO-A和IFN溶液的制备和实施例3相同。 The same preparation of the polymer solution and IFN PO-A, and Example 3. 然后加水稀释至聚合物PO-A的浓度为15 mg/g(IFN的浓度应为0.188 mg/g)。 Then diluted with water to a polymer concentration of PO-A 15 mg / g (IFN concentration should be 0.188 mg / g). 通过0.2pm的滤膜过滤后,溶液中加入1NHCl酸化至pH:4。 After 0.2pm by membrane filtration, was added 1NHCl was acidified to pH: 4. 所得溶液是乳白色的。 The resulting solution was milky white.

酸化的聚合物-IFN溶液的雾化 -IFN atomized polymer solution was acidified

溶液的雾化条件和实施例3相同。 Atomized solution and the same conditions of Example 3.

所得微颗粒的表征: The resulting microparticles characterization:

所得颗粒的尺寸(根据T0试验)为D50 = 5 pm。 The resulting particle size (according to test T0) of D50 = 5 pm.

将粉末于雾化浓度分散于水中以后,IFN的HPLC检测和雾化前溶液的检测相同,未发现分解形式。 The atomized powder is dispersed in water after concentration, the same is detected by HPLC the solution prior to atomization of IFN and found no decomposition form.

实施例6:用PVP包膜并含有IFN-a2b的聚合物PO-A的干燥微米-尺寸的颗粒 Example 6: coated with IFN-a2b and containing PVP polymer PO-A drying micron - particle size

的制备 Preparation

按照实施例3使用的方案从PO-A/IFN混合物获得由聚合物PO-A和IFN的干燥微颗粒形成的粉末。 According to the protocol used in Example 3 to obtain a powder formed from the polymer PO-A and IFN from the dried microparticles PO-A / IFN mixture.

在雾化步骤结束后,1.5 g这种粉末再悬浮于含有7 g/1 PVP K30型注射级的聚乙烯吡咯烷酮(PVP)的乙醇溶液中。 At the end of the atomization step, 1.5 g of this powder was suspended in an ethanol solution containing 7 g / 1 PVP K30 injectable grade polyvinylpyrrolidone (PVP) in the. 该乙醇悬浮液搅拌2h,然后在配备了有机溶剂提取捕集器(-20。C惰性循环)的Biichi B290型的喷雾干燥装置中再雾化一次以及在氮气条件下密闭循环操作。 The ethanol suspension was stirred for 2h, and then organic solvent extraction with a trap (-20.C inert cycle) Biichi B290-type spray drying apparatus and then atomized again sealed under nitrogen cycle of operation. 以5 ml/min的速度抽出液体溶液,并通过供氮(7bar-670 1/h)的喷嘴雾化。 At a rate of 5 ml / min was withdrawn liquid, and by supplying nitrogen (7bar-670 1 / h) of the atomizing nozzle. 抽出流速(干空气)为40 m3/h。 Extraction flow rate (dry air) of 40 m3 / h. 进口温度维持在8(TC,这些条件下的出口温度为55X:。 所得微颗粒的表征: Inlet temperature was maintained at 8 (TC, characterization of the resulting microparticles 55X :. outlet temperature under these conditions:

所得颗粒的尺寸(根据T0试验)为D50 = 6 The resulting particle size (according to test T0) of D50 = 6

将粉末于雾化浓度分散于水中以后,IFN的HPLC检测和雾化前溶液的检测相同,未发现分解形式。 The atomized powder is dispersed in water after concentration, the same is detected by HPLC the solution prior to atomization of IFN and found no decomposition form.

实施例7:含有IFN-a2b的聚合物P0-B的干燥微米-尺寸的颗粒的制备聚合物-IFN溶液的制备和雾化方法和实施例3相同,其中聚合物PO-A用聚合物PO-B代替。 Example 7: IFN-a2b dried microns containing polymer P0-B - the same preparation and atomization process and the polymer prepared in Example -IFN solution particle size 3, wherein the polymer PO-A polymer PO -B instead.

所得微颗粒的表征: The resulting microparticles characterization:

所得颗粒的尺寸(根据T0试验)为D50 = 4 The resulting particle size (according to test T0) of D50 = 4

将粉末于雾化浓度分散于水中以后,IFN的HPLC检测和雾化前溶液的检测相同,未发现分解形式。 The atomized powder is dispersed in water after concentration, the same is detected by HPLC the solution prior to atomization of IFN and found no decomposition form.

实施例8:含有hGH的聚合物PO-A的干燥微米-尺寸的颗粒的制备浓縮的hGH溶液的制备: Example 8: Polymer PO-A was dried microns hGH containing a - Preparation of the particle size of the concentrated hGH solution:

60 g重组人生长激素溶液(浓度3.9 mg/g)于25。 60 g of recombinant human growth hormone solution (a concentration of 3.9 mg / g) at 25. C解冻1 h 30,然后通过10 kD滤膜以正向超滤(frontal ultrafiltration)的方式浓縮约6倍直到浓度达到24 mg/g(HPLC监控)。 C Thaw 1 h 30, then by 10 kD ultrafiltration membrane in a forward manner (frontal ultrafiltration) about 6-fold concentrated until the concentration reached 24 mg / g (HPLC monitoring).

和聚合物溶液的混合物 A mixture of polymer solution and

将28 g水加入到52 g浓縮的浓度为30 mg/g的聚合物PO-A的溶液中将其稀释至19.5 mg/g。 The water was added to 28 g in the 30 mg / g polymer PO-A was diluted to 19.5 mg / g concentrated to a concentration of 52 g. 将8 g浓度为24 mg/g的hGH溶液缓慢倒入稀释后的聚合物溶液中。 The concentration of 8 g 24 mg / g of hGH solution was slowly poured into the diluted polymer solution. 混合物于室温下静置过夜然后用0.2 pm除菌滤膜过滤。 The mixture was allowed to stand at room temperature overnight and then filtered through 0.2 pm filter sterilization.

聚合物/hGH溶液的雾化 Atomizing the polymer / hGH solution

溶液的雾化条件和实施例3相同。 Atomized solution and the same conditions of Example 3.

所得微颗粒的表征: The resulting microparticles characterization:

所得颗粒的尺寸(根据T0试验)为D50 = 5 The resulting particle size (according to test T0) of D50 = 5

将粉末于雾化浓度分散于水中以后,hGH的HPLC检测和雾化前溶液的检测相同,未发现分解形式。 The atomized powder is dispersed in water after concentration, the detection and the detection of hGH by HPLC the solution prior to atomization same, No decomposition form.

45实施例9:含有IL-2的聚合物PO-A的干燥微米-尺寸的颗粒的制备浓縮的IL-2溶液的制备: Example 945: containing the polymer PO-A IL-2 is dried micron - particle size Preparation of IL-2 concentrated solution:

100 g用SDS稳定的浓度为2 g/1的IL-2冷冻溶液于室温下解冻,然后该溶液冷却到0-2°C。 100 g stabilized with SDS concentration of 2 g / 1, IL-2 frozen solution was thawed at room temperature and then the solution was cooled to 0-2 ° C. 100 g无水乙醇缓慢加入到该溶液中以沉淀蛋白质。 100 g of anhydrous ethanol was slowly added to the solution to precipitate the protein. 沉淀物用0.65/0.45 nm的膜在正向渗滤池(frontal diafiltration cell)中以过滤的方式回收并用200 g水洗涤。 The precipitate was washed with 0.65 / 0.45 nm filter membrane in a manner diffusion cells recovered in the forward (frontal diafiltration cell) and washed with 200 g of water. 沉淀物用氮气流干燥,然后溶解于10 g预先冷却(〈5")的0.02 N 氢氧化钠中以获得浓度为20mg/g、 pH42并且无SDS的澄清蛋白质溶液。溶液用HPLC方法进行严格检测。 The precipitate was dried with a nitrogen stream, then dissolved in 10 g pre-cooled (<5 ") of 0.02 N sodium hydroxide to obtain a concentration of 20mg / g, pH42 and no SDS in a clear protein solution. The solution was rigorous testing using HPLC method .

和聚合物溶液的混合物 A mixture of polymer solution and

将39 g水加入到96 g浓縮的(浓度为30mg/g)实施例l的聚合物PO的溶液中将其稀释。 The 39 g of water was added to 96 g diluted in the concentrated solution of polymer PO (concentration of 30mg / g) Example l embodiment. 将9 g前面的浓度为20 mg/g的IL-2溶液缓慢倒入稀释后的聚合物溶液中。 The foregoing concentration of 9 g 20 mg / g of the IL-2 solution was slowly poured into the diluted polymer solution.

含有20 mg/g聚合物PO和1.25 mg/g IL-2的混合物于室温下静置过夜然后用0.2 pm除菌滤膜过滤。 20 mg / 1.25 mg g polymer mixture and PO / g IL-2 containing allowed to stand overnight at room temperature then treated with 0.2 pm sterilizing membrane filter.

聚合物-IL-2溶液的雾化溶液的雾化条件和实施例3相同。 Atomizing the same conditions as in Example atomization of the solution -IL-2 polymer 3 solution. 所得微颗粒的表征: The resulting microparticles characterization:

所得颗粒的尺寸(根据T0试验)为D50 = 5 The resulting particle size (according to test T0) of D50 = 5

将粉末于雾化浓度分散于水中以后,IL-2的HPLC检测和雾化前溶液的检测相同,未发现分解形式。 The atomized powder is dispersed in water after concentration, detected by HPLC and the same solution prior to atomization of IL-2, it was not found in exploded form.

实施例10:含有胰岛素的聚合物PO-A的干燥微米-尺寸的颗粒的制备40 g浓度为20.3 mg/g的胰岛素溶液的制备: Example 10: insulin-containing polymer was dried microns of PO-A - 40 g particle size prepared at a concentration of insulin solution to prepare 20.3 mg / g of:

0.83g含水量2.5%、活性为28.7 Ul/g的重组人胰岛素(粉末)加入到玻璃烧瓶中。 0.83g water content of 2.5%, an activity of 28.7 Ul / g recombinant human insulin (powder) was added to a glass flask. 加入23.62 g水,然后胰岛素用缓慢的磁力搅拌分散。 23.62 g of water was added, and then stirred with slow insulin magnetic dispersion. 加入6.22g 0.1N HC1 直到获得澄清的酸性胰岛素溶液。 Was added until acidic 6.22g 0.1N HC1 solution to obtain a clear insulin. 然后加入9.32 g 1N氢氧化钠以获得pH 7-8 的最终澄清液。 Was then added 9.32 g 1N sodium hydroxide to obtain a pH 7-8 in the final supernatant. 该胰岛素溶液用0.2 pm除菌滤膜过滤。 The insulin solution was filtered through a 0.2 pm filter sterilization.

和聚合物溶液的混合物 A mixture of polymer solution and

将37.66 g前面获得的浓縮胰岛素溶液缓慢倒入(同时磁力搅拌)220 g 30 mg/g的聚合物PO溶液中。 37.66 g of the obtained concentrated insulin solution was slowly poured into the front (magnetic stirring) 220 g 30 mg / g solution of polymer PO. 然后往溶液中加入72.34 g水。 Was then added to the solution 72.34 g of water. 混合物于室温下静置4小时然后用0.2 iLim滤膜过滤。 The mixture was allowed to stand at room temperature for 4 hours then treated with 0.2 iLim membrane filter.

聚合物-胰岛素溶液的雾化 Polymer - atomized insulin solution

该溶液然后在Biichi B290型的喷雾干燥装置上雾化。 The solution is then atomized on a Biichi B290-type spray-drying apparatus. 以5 ml/min的速度抽出液体溶液,并通过供氮(7 bar-900 1/h)的喷嘴雾化。 At a rate of 5 ml / min was withdrawn liquid, and by supplying nitrogen (7 bar-900 1 / h) of the atomizing nozzle. 抽出流速(干空气)为40 m3/h。 Extraction flow rate (dry air) of 40 m3 / h. 进口温度维持在120。 Inlet temperature was maintained at 120. C,这些条件下的出口温度为70。 C, outlet temperature of 70 under these conditions. C。 C.

聚合物-胰岛素溶液的雾化 Polymer - atomized insulin solution

溶液的雾化条件和实施例3相同。 Atomized solution and the same conditions of Example 3.

所得微颗粒的表征: The resulting microparticles characterization:

所得颗粒的尺寸(根据T0试验)为D50 = 5 The resulting particle size (according to test T0) of D50 = 5

将粉末于雾化浓度分散于水中以后,胰岛素的HPLC检测和雾化前溶液的检测相同,未发现分解形式。 The atomized powder is dispersed in water after concentration, the detection of insulin by HPLC and the same solution prior to atomization, No decomposition form.

实施例ll:根据本发明所获得的微颗粒的特征:尺寸和稳定性 Example ll embodiment: According to a feature of the obtained microparticles of the present invention: Size and Stability

微颗粒的尺寸根据TO试验测定,稳定性根据S1试验测定。 The size of the microparticles TO test measures, determined according to the stability test S1. 蛋白质的分解通过HPLC比较雾化前后的色谱图进行评估。 The decomposition of protein was assessed by comparing HPLC chromatograms before and after atomization.

实施例 根据TO试验的D50尺寸(pm) 经过根据ST1的稳定性试验后的D50尺寸((im) 雾化所导致的蛋白质分解* The TO Test Example D50 size (PM) through D50 according to the size of the stability test ST1 ((IM) caused by atomization proteolytic *

3 5 5 无 355 free

4 5 5 无 455 free

5 5 5 无 555 free

6 6 5 无 665 free

7 4 5 无 745 free

8 5 5 无 855 free

9 5 5 无 955 free

10 5 5 无 1055 None

*无表示没有观察到分解形式。 * No means that no decomposition was observed form.

结果表明在两性分子聚氨基酸存在下雾化蛋白质能够保持蛋白质的稳定性。 The results show that the presence of amphiphilic poly amino acid protein can be maintained at an atomizing protein stability. 在固体形式时,根据所描述的方案,粉末是稳定的。 In solid form, according to the described embodiment, the powder is stable. 很显然并且可以预期 It is clear and can be expected

粉末在5"C时其稳定性可达至少两年。 Powder stability at 5 "C up to at least two years.

雾化是一种可能引起蛋白质分解的方法。 Atomizing method may cause a breakdown of proteins. 通过HPLC分析比较雾化前后的色谱图显示没有观察到分解的形式。 By comparing the chromatogram of HPLC analysis showed no fogging was observed before and after exploded form.

47在两性分子聚氨基酸存在下雾化蛋白质可以得到长时间稳定的微颗粒而且不会引起蛋白质的分解。 47 in the presence of acid poly amphiphilic proteins may be atomized stably obtained for a long time and the microparticles do not cause decomposition of protein.

这些粉末可以固体形式使用(例如吸入、在压力下通过枪头进行无针头注射) 或者在合适的介质中重构后以可注射悬浮液的形式使用。 These powders can be used in solid form (e.g. inhalation, a needle-free injection under pressure through the pipette tip), or after reconstitution in a suitable medium may be used in the form of an injectable suspension.

实施例12:从实施例3-10的粉末开始悬浮液的重构 Example 12: Starting from the reconstituted powder of Example 3-10 embodiment suspension

根据进行DP1试验所描述的方案,粉末在三种不同的介质中进行重构: The scheme described DP1 test, powder was reconstituted in three different media:

A. pH 7.4的磷酸盐缓冲盐水溶液 A. pH 7.4 phosphate buffered saline solution

BOIM的氯化镁水溶液 BOIM magnesium chloride aqueous solution

C. 癸酸甘油三酯溶液(Miglyo1⑧812) 然后通过进行以下步骤评估样品: C. capric triglyceride solution (Miglyo1⑧812) samples were then evaluated by performing the following steps:

-在前面提及的一种介质中分散-转移到注射器中然后通过25G针头注射。 - dispersion in a medium in the aforementioned - transferred into a syringe and injected through a 25G needle. 如果通过25G针头吸取/注射能够回收至少80n/。 If a 25G needle by suction / injection can be recovered at least 80n /. 的悬浮液,则认为其具有良好的性质。 The suspension is considered to have good properties.

结果:<table>table see original document page 48</column></row> <table> Results: <table> table see original document page 48 </ column> </ row> <table>

在稳定性试验期间还检测到蛋白质在介质A、 B和C中重构后不发生分解。 During further stability test detected no decomposition after protein reconstituted in medium A, B and C.

这些结果显示只有实施例7的组合物才能获得水中用PBS缓冲的pH=7.4的稳定的和可注射的悬浮液。 These results show that only the composition of Example 7 in water to obtain a pH = 7.4 buffered with PBS stable and injectable suspensions. 对于大部分实施例(5除外),含有二价盐的重构介质能够改善这些性质。 For most embodiments (excluding 5), divalent salt-containing reconstituted medium capable of improving these properties. 在Miglyol介质中,所有的制剂都易于分散、注射并且是稳定的。 In Miglyol medium, all formulations are easily dispersed, and stable injection.

根据所用的聚合物并不能容易地获得这些性质的组合。 The polymer used can not be easily obtained and a combination of these properties. 发明人的贡献在于,经过多次试验,发现了某些赋形剂和重构介质能够使其具有这些性质。 Contribution to the inventors that, after several tests, and found that certain excipients reconstituted medium capable to have these properties. 实施例13:重构制剂粘度的测定 Example 13: Determination of the viscosity of reconstituted formulation

30 mg/ml前面实施例的介质B和C的粉末的分散体(介质B中的实施例5除外) 能够得到稳定的低粘度悬浮液;在介质B中测得的粘度均低于IO mPa.s,在介质C中测得的粘度都约在30-40 mPa.s。 Powder dispersion medium B and C of Example 30 mg / ml foregoing (medium B in Example 5 with the exception) can be obtained a stable, low viscosity of the suspension; B, in the medium were lower than the viscosity measured IO mPa. s, C in the medium are measured viscosity of about 30-40 mPa.s.

经过比较,根据现有技术的纳米颗粒形式的相同聚合物的组合物在这个浓度(30 mg/ml)下不能获得可接受的粘度(〈100 mPa.s)。 By comparison, according to the same polymer composition in the form of nano-particles of the prior art in this concentration (30 mg / ml) can not obtain an acceptable viscosity (<100 mPa.s) lower.

实施例14:含有IFN-a2b的聚合物P0-A的干燥微米-尺寸的颗粒的冻干粉末 Example 14: IFN-a2b dried microns containing polymer P0-A - the lyophilized powder particle size

形式的制备 Prepared in the form of

首先按照实施例3的描述制备颗粒。 First, preparation of the particles according to the embodiment described in Example 3. 雾化步骤在无菌条件下进行,并且颗粒在无菌容器中回收。 The atomization step is carried out under sterile conditions, and the particles recovered in a sterile container.

在M^+离子存在下颗粒的再分散 ^ + Ions under the redispersed particles M

在雾化阶段后,颗粒立即回收并且在无菌条件下再分散到Ol MMgCl2水溶液中。 After the atomization phase, the particles immediately recovered and redispersed under aseptic conditions into an aqueous solution Ol MMgCl2. 调整所加入的MgCl2溶液的量,使悬浮液中PO-A聚合物的浓度为大约为40 mg/ml。 Adjusting the amount of MgCl2 solution added, the concentration of the suspension PO-A polymer is about 40 mg / ml. 加入l N氢氧化钠调节pH值为6.5。 L N sodium hydroxide solution was added adjust the pH 6.5.

冻干 Lyophilized

悬浮液分装在Lyoguard⑧型的容器中,其能够使悬浮液在冻干期间保持无菌:容器然后于无菌条件下在实验室冻干仪器(实验室桌面单位Christ, Osterode, Germany)上循环冻干72 h。 The suspension was dispensed in Lyoguard⑧ type container, which can be kept sterile in the suspension during lyophilization: circulation vessel was then lyophilized on a laboratory instrument (laboratory benchtop unit Christ, Osterode, Germany) under sterile conditions lyophilized 72 h. 粉末在使用前在无菌条件下分装在烧瓶中。 Powder before use under sterile conditions dispensed in a flask.

实施例15:从实施例3获得的微颗粒粉末和实施例14的微颗粒粉末重构的对 Example 15: micro particles obtained from Example 3 and Example 14 embodiment the micro-particles of the reconstructed

ratio

对于该对比,两种情况的重构悬浮液的体积相同(大约l ml),并且重构的烧瓶也相同(3 ml玻璃烧瓶)。 For this comparison, the same volume of reconstituted suspension in both cases (about l ml), and reconstructs the same flask (3 ml glass flask). 调节粉末的量使两种悬浮液都最终含有40 mg/ml的聚合物和0.5 mg/ml的IFN-a2b。 Adjusting the amount of the powder so that the final two suspensions are polymers containing 40 mg / ml and 0.5 mg / ml of IFN-a2b. 实施例3的粉末在0.1 M MgCl2水溶液中重构,实施例14的粉末(其已经含有Mg^离子)在纯水中重构。 Reconstituted powder of Example 3 in 0.1 M MgCl2 aqueous solution, the powder of Example 14 (which has been ion-containing Mg ^) reconstituted in purified water.

第一次,烧瓶用人工搅拌。 First, the flask was manually stirred. 当实施例3的粉末需要至少15分钟来再分散时, 实施例14的粉末分散更快,在少于5分钟的时间获得了均匀的悬浮液。 When the powder of Example 3 at least 15 minutes and then dispersing the powder of Example 14 dispersed quickly, in less than 5 minutes to obtain a homogeneous suspension. 然后在两 Then two

49烧瓶中都插入磁力搅拌棒,以相同的方式搅拌lh。 49 are inserted into a flask with a magnetic stir bar, is stirred in the same manner lh.

该搅拌结束后,比较两种悬浮液的特征:两种悬浮液的颗粒尺寸和粘度相似。 After completion of the stirring, wherein the comparison of two suspension: similar particle size and viscosity of the two suspensions. <table>table see original document page 50</column></row> <table>实施例16:皮下注射基于两性分子聚氨基酸微颗粒形式的制剂后,狗中的 <Table> table see original document page 50 </ column> </ row> <table> Example 16: subcutaneous injection of microparticles in the form of a polyamino acid formulations based on amphiphilic molecules, dogs

IFN药代动力学 IFN Pharmacokinetics

用以下制剂处理8条Beagle小猎犬(重量9 ± 0.6 kg): 8 beagle dogs Beagle processing (by weight 9 ± 0.6 kg) with the following formulation:

<table>table see original document page 50</column></row> <table>IFN IR (IR指立即释放)对应于重组人干扰素溶液(PCGen,批号IB05.0516), 其浓度、pH和摩尔渗透压浓度调节到一定值([IFN] = 0.5 mg/ml、 pH = 6.5和354 mOsm)。 <Table> table see original document page 50 </ column> </ row> <table> IFN IR (IR refers to immediate release) corresponding to the recombinant human interferon solution (PCGen, Lot IB05.0516), concentration, pH, and moles osmolarity adjusted to a certain value ([IFN] = 0.5 mg / ml, pH = 6.5 and 354 mOsm).

制剂1的颗粒按照实施例14从相同PCGen批次的干扰素制备:冻干粉末于无菌条件下(在层流气流罩下)按照和实施例15中的描述相类似的方法再悬浮于水中。 Particles of the formulation according to Example 1 14 Preparation of interferon from the same batch PCGen: lyophilized powder under sterile conditions (in the laminar air flow hood) and described in accordance with Example 15 Similar procedure as described resuspended in water .

药代动力学结果汇总于下表中: The pharmacokinetic results are summarized in the following table:

<table>table see original document page 50</column></row> <table>其中- <Table> table see original document page 50 </ column> </ row> <table> wherein -

-Cmax是最大血清IFN浓度: -Cmax is the largest concentration of serum IFN:

-T 〉 50 pg/ml是血清IFN浓度大于5 ng/ml的时间; -AUC代表血清IFN浓度作为时间函数的曲线下面积-RBA代表相对于速释制剂的生物利用度; -Ts。 -T> 50 pg / ml serum IFN concentration greater than 5 ng / ml of time;-AUC serum IFN concentrations representative of the area as a function of time is represented by the following -RBA curve relative bioavailability of immediate release formulation; -Ts. ^ue代表盐析总IFN的50。 Representative ^ ue salting total of 50 IFN. /。 /. 所需的时间。 The time required. IFNIR具有速释曲线,其最大血清浓度为25.2士0.4ng/ml,在5h的中位时间后获得(范围:3-5 h)。 IFNIR having immediate release profile, a maximum serum concentration of 25.2 Disabled 0.4ng / ml, obtained after a median time of 5h (range: 3-5 h). 24小时以后循环IFN检测不到。 After 24-hour cycle IFN undetectable.

制剂1提供了IFN的药代动力学曲线的主要修正,其非常缓慢地释放,在60h 的中位时间(范围:48-96 h)后获得最大血清浓度0.5 ± 0.2 ng/ml (低于IR形式的最大血清浓度50倍)。 Formulation 1 provides a corrected main pharmacokinetic profile of IFN, which is very slowly released in 60h median time (range: 48-96 h) maximum serum concentration of 0.5 ± 0.2 ng / ml after (lower than the IR the maximum serum concentration in the form of 50 fold). 药代动力学的总体趋势是假稳态形式的非常平坦的曲线。 Pharmacokinetics of the overall trend is very flat curve false steady-state form. 循环IFN的水平在192-240小时(8-10天)回到检测不到的浓度。 In the levels of circulating IFN 192-240 hours (8-10 days) back to undetectable concentration. 该制剂具有较低的AUC:相对生物利用度损失66% (RBA = 34%),其T5w高出IFN IR的丁5()%31]£;大约20倍。 This formulation has a lower AUC: lost relative bioavailability of 66% (RBA = 34%), which is higher than T5w D 5 ()% 31 IFN IR of] £; about 20 times.

实施例17:皮下注射基于两性分子聚氨基酸微颗粒形式的制剂后,狗中的胰岛素药代动力学 Example 17: preparation of polyamino acid after subcutaneous injection in the form of microparticles, dog pharmacokinetics of insulin based on amphiphilic molecules

本实施例的对比物Lantus⑧是一种改性的胰岛素类似物(甘精胰岛素-Sanofi Aventis)。 Comparative composition Lantus⑧ embodiment of the present embodiment is a modified insulin analog (insulin glargine -Sanofi Aventis). 在人胰岛素的一级结构上改变了两个氨基酸赋予Lantus⑧延长释放性质,其释放超过24h时间,其通过注射位点的沉淀物实现。 In the primary structure of human insulin two amino acid changes Lantus⑧ impart extended release properties that releases over time 24h, by the site of injection achieved precipitate.

一组6条健康的Beagle犬(体重:11.4 ± 1 kg)在两阶段交叉试验中用制剂Lantus⑧进行处理, 一组8条健康的Beagle犬(体重:11.8 ± 1 kg)在四阶段交叉试验中用制剂2处理,二对二。 A group of six healthy Beagle dogs (body weight: 11.4 ± 1 kg) in a crossover study with formulations Lantus⑧ two stage process, a group of eight healthy Beagle dogs (body weight: 11.8 ± 1 kg) in a four-stage cross-Test was treated with formulation 2, two pairs of two. 下表对处理方法进行了简述: The following table processing methods are briefly:

狗的数目 [胰岛素] (賜 [PO] (mg/g) ,g) 体积剂量(Hl/kg) The number of dogs [insulin] (giving [PO] (mg / g), g) Volume dose (Hl / kg)

Lantus® (批号:40N300) 12 100 0 1 10 Lantus® (Lot: 40N300) 12 100 0 1 10

制剂2 8 100 30 1 10 Formulation 2810030110

制剂2的颗粒按照实施例10制备,目标比例是3.5 mg (100 IU)胰岛素用30 mg PO聚合物。 Particles of the formulation 2 prepared in Example 10, a target ratio is 3.5 mg (100 IU) of insulin with 30 mg PO polymer. 制剂通过将雾化的粉末分散在Ol M的MgCl2中并且磁力搅拌悬浮液lh进行制备。 By atomized powder formulation dispersed in MgCl2 Ol M and the suspension was magnetically stirred for lh prepared. 制剂的pH是6.2,摩尔渗透压浓度为348mOsm。 pH of the formulation is 6.2, osmolality 348mOsm. 对应的以I^试验测定的颗粒尺寸为D50 = 12 pm,制剂在2(TC时的粘度大约为5 mPa.s。 I ^ corresponding to the experimentally determined particle size D50 = 12 pm, the formulation viscosity (TC 2 when approximately 5 mPa.s.

血糖在血液生化分析仪(Advia 1650, Bayer Diagnostics)上用酶方法(己糖激 Glucose in the blood biochemical analyzer (Advia 1650, Bayer Diagnostics) using an enzyme method (hexokinase

酶)计量。 Enzyme) measured.

药代动力学结果的分析基于相对于时间的基础血糖的百分比。 Kinetic analysis of the results with respect to pharmacokinetic percentage basis based on the time of blood glucose. 下表汇总了药代动力学数据: The following table summarizes the pharmacokinetic data:

51<table>table see original document page 52</column></row> <table> 51 <table> table see original document page 52 </ column> </ row> <table>

其中: among them:

-C曲是己观察到的基础血糖的最小百分比; -C song is the minimum percentage basis of blood glucose had observed;

-八? -Eight? 00).3611(血糖曲线面积百分数)代表在剂量后0到3611的时间 00) .3611 (area percent glucose curve) represents the time after the dose of 0-3611

期间,基础血糖(100%)和代表血糖进程(以基础血糖的百分数表示) During the basis of blood glucose (100%) and represents the process of blood glucose (blood sugar expressed as a percentage basis)

AAi rHi A4i ~^ ,、;tT iVi古T7^T CTJ CO] 心I 口J PTJ衣即 AAi rHi A4i ~ ^ ,,; tT iVi ancient T7 ^ T CTJ CO] Heart J PTJ clothing i.e., port I

-T5o。 -T5o. /。 /. ATOc代表获得50Q/。 On behalf of ATOc obtain 50Q /. 的APGCo.36h所需的时间。 The time required to APGCo.36h.

参照物Lantus⑧给药后血糖从第一小时起迅速减少。 After administration Lantus⑧ reference blood glucose rapidly reduced from the first hour. 然后甘精胰岛素的低血糖行为保持18至36小时的时期(平均在30小时后血糖回到其基础水平)。 Then insulin glargine hypoglycemia behavior holding period of 18-36 hours (average blood glucose returns to its basal level after 30 hours).

作为比较,制剂2给药后血糖也从第一小时起迅速减少。 As a comparison, after administration of formulation 2 quickly reduce blood glucose from the first hour. 然后,基础血糖的百分比平均维持稳态直到36小时。 Then, the average percentage based glucose homeostasis is maintained until 36 hours. 制剂l得到的C^平均明显高于参照物Lantus⑧ 得到的值,这能够可观地减少糖尿病病人的严重低血糖现象。 C ^ l formulation obtained values ​​were higher than reference Lantus⑧ obtained, which can considerably reduce the phenomenon of severe hypoglycaemia in diabetic patients.

制剂2的作用时间明显大于长效作用的参照物Lantus⑧的作用时间。 Formulation 2 reaction time is substantially greater than the reference Lantus⑧ long acting duration of action. 这由制剂2较高的平均T50。 This average T50 Formulation 2 by a high. /。 /. APGc值证实。 APGc value confirmed.

与参照物Lantus⑧相比,制剂2的玎00).3611损失仅为24%。 Compared with the reference Lantus⑧, preparation Ding 00) .3611 2 loss of only 24%.

Claims (41)

1.一种聚合物(PO)的微颗粒,其含有至少一种活性成分(AP),所述聚合物PO ●是带有疏水性基团(HG)和亲水性基团的水溶性生物可降解的两性分子(共)聚合物, ●在pH 7.0,等渗条件下,在水中自发地形成纳米颗粒的胶体悬浮液, ●并且和所述AP非共价地结合; 所述微颗粒a.通过雾化PO的溶液或胶体悬浮液获得,所述PO的溶液或胶体悬浮液含有至少一种AP, b.以T试验测得的尺寸为0.5-100微米,优选1-70微米,更优选2-40微米, c.并且在“可分散性”试验DP1中在胶体悬浮液中是可分散的。 A polymer (PO) microparticles which contain at least one active ingredient (the AP), the polymer is water soluble, biologically PO ● having a hydrophobic group (HG) and a hydrophilic group amphiphilic biodegradable (co) polymer, ● at pH 7.0, isotonic condition, formation of nanoparticles in colloidal suspension in water spontaneously, and ● the AP and non-covalently bound; a said microparticles by PO atomizing a solution or colloidal suspension is obtained, the PO solution or colloidal suspension containing at least one AP, b. a T-test as measured in a size of 0.5 to 100 microns, preferably 1-70 microns, more preferably 2-40 microns, c. and "dispersible" in the test DP1 colloidal suspension is dispersible.
2. 如权利要求1所述的微颗粒,其在ST1试验或ST2试验中是稳定的。 The microparticles as claimed in claim 1, which is stable in the test ST1 or ST2 test.
3. 如权利要求1或2所述的微颗粒,其中PO是嵌段型或者无序型的共聚物。 Or the microparticles as claimed in claim 12, wherein PO is a block type or a random type copolymer.
4. 如前述任何一项权利要求所述的微颗粒,其中所述PO的亲水性基团是至少部分电离的可电离基团(IG)。 4. any of the preceding claims said microparticles, wherein PO is the hydrophilic group is at least partially ionized ionizable group (IG).
5. 如前述任何一项权利要求所述的微颗粒,其中所述聚合物PO是两性分子(共) 聚氨基酸或两性分子(共)聚氨基酸的混合物。 5. A microparticle according to any one of claims the preceding claims, wherein said amphiphilic polymer PO is a (co) poly-amino acid molecule, or amphoteric (co) polyamino acids.
6. 如前述任何一项权利要求所述的微颗粒,其中PO是聚氨基酸,其主链由天冬氨酸单元和/或谷氨酸单元形成,这些单元中的至少一部分通过在链中或者链端接枝至少一个疏水性基团(HG)而改性。 6. any of the preceding claims microparticles, wherein PO is a polyamino acid, which backbone is formed by aspartic acid units and / or glutamic units, at least a portion of these units in the chain or by branched chain termination least one hydrophobic group (HG) and modified.
7. 如前述任何一项权利要求所述的微颗粒,其中PO由下列通式(I)(基团-COOR3 包括羧基和RS之间的键是离子键-COCT+R3的形式)定义:<formula>formula see original document page 2</formula>其中:■ R'代表H、 (:2到do的直链垸基或C3到do的支链垸基、苄基或-R4-[HG];■ NHR'是末端氨基酸残基;■ 112代表H、 C2到d。的直链酰基或Q到C,。的支链酰基或-R4-[HG];■ RS是末端焦谷氨酸根残基;■ R3是H,或者■ +&3优选自包括以下的组:-金属阳离子,优选自包括钠、钾、钙和镁的亚组, -有机阳离子,优选自包括以下的亚组:• 基于胺的阳离子,• 基于低聚胺的阳离子,• 基于聚胺的阳离子(特别优选聚乙烯亚胺),• 基于氨基酸的阳离子,优选自包括基于赖氨酸或精氨酸的阳离子的组,-或者阳离子聚氨基酸,优选自包括聚赖氨酸或低聚赖氨酸的亚组;■ W代表直接键或基于1到4个氨基 7. A microparticle according to any of the preceding claims, wherein the PO by the following formula (I) (group -COOR3 include carboxyl groups and the bond between the ionomer is RS form -COCT + R3) is defined: < formula> formula see original document page 2 </ formula> where: ■ R 'representative of H, (: 2 to do a straight-chain alkyl or C3 to do branched chain alkyl, benzyl or -R4- [HG]; ■ NHR 'is a terminal amino acid residue; ■ 112 representative of H, C2 to d or a straight-chain acyl group of Q to C ,. branched acyl or -R4- [HG];. ■ RS is a root terminal pyroglutamate residue ; ■ R3 is H, or ■ + & 3 is preferably selected from the group comprising: - metal cations, preferably from the group comprising an alkylene group of sodium, potassium, calcium and magnesium, - organic cations, preferably from the group comprising the following sub-groups: • amine-based cations, • cations based on oligoamine, • the cations based on polyamine (polyethyleneimine is particularly preferred), • cations based on amino acids, preferably from the group consisting of cations based on lysine or arginine group, - or cationic polyamino acid, preferably from the subgroup comprising polylysine or oligomeric lysine; ■ W represents a direct bond or a 1 to 4-amino-based 残基的间隔子;■ 独立地,A代表-CH2-基团(天冬氨酸单元)或-CH2-CH2-基团C谷氨酸单元);■ n/(n+m)是摩尔接枝率,在pH为7, 25°C时溶于水中PO的摩尔接枝率值足够低,以形成PO亚微米尺寸颗粒的胶体悬浮液;n/(n+m)优选为1-25 mol%,更优选1-15 mol%;■ n+m是聚合度,其为IO-IOOO,优选50-300;■ HG代表含有6-30个碳原子的疏水性基团。 Spacer residues; ■ independently, A represents a group -CH2- (aspartic unit) or -CH2-CH2- group C glutamic unit); ■ n / (n + m) is connected mol rate branches, at pH 7, 25 ° C when dissolved in water PO molar grafting rate value low enough to form a colloidal suspension of submicron sized particles of PO; n / (n + m) is preferably 1-25 mol %, more preferably 1-15 mol%; ■ n + m is the degree of polymerization, which is IO-IOOO, preferably 50-300; ■ HG represents an hydrophobic group of 6-30 carbon atoms.
8.如前述任何一项权利要求所述的微颗粒,其中一种或多种PO对应于以下通式(11)、 (ni)和(IV)(基团-COOI^包括羧基和RS'之间的键是离子键-COCT+RS'的形式) 中的一种:<formula>formula see original document page 4</formula>■ HG代表含有6-30个碳原子的疏水性基团;■ R^是直链的二价CVC6亚烃基链;■ R3'是H,或者■ +尺3'优选自包括以下的组:-金属阳离子,优选自包括钠、钾、钙和镁的亚组, -有机阳离子,优选自包括以下的亚组:• 基于胺的阳离子,• 基于低聚胺的阳离子,• 基于聚胺的阳离子(特别优选聚乙烯亚胺),• 基于氨基酸的阳离子,优选自包括基于赖氨酸或精氨酸的阳离子的组,-或者阳离子聚氨基酸,优选自包括聚赖氨酸或低聚赖氨酸的亚组;■ R^是直链的二价CrQ亚烃基链,其中一个或两个亚甲基单元,优选位于R5Q每个末端的亚甲基单元,可以独立地被 Said microparticles (ni) and (IV) (including carboxyl groups -COOI ^ and RS '8. Any of the preceding claims, wherein the one or more PO corresponding to the following general formula (11), ionic bond is the bond between -COCT + RS 'form) one of: <formula> formula see original document page 4 </ formula> ■ HG represents an hydrophobic group of 6-30 carbon atoms; ■ R ^ is a straight chain divalent CVC6 alkylene chain; ■ R3 'is H, or ■ + ft 3' is preferably selected from the group comprising: - metal cations, preferably from the group consisting of sodium, potassium, calcium and magnesium, subgroup - an organic cation, preferably from the group comprising the following sub-groups: • cations based on amine, • the cations based on oligoamine, • the cations based on polyamine (polyethyleneimine is particularly preferred), • cations based on amino acids, preferably from the group consisting based lysine or arginine cationic group, - or cationic polyamino acids, preferably from the subgroup comprising polylysine or oligomeric lysine; ■ R ^ is a straight chain divalent CrQ alkylene chain, wherein one or two methylene units, preferably located at each end R5Q methylene units may independently be -0或-NH代替;■ W代表直接键或基于1到4个氨基酸残基的间隔子;麵独立地,A代表-CH2-基团(天冬氨酸单元)或-CH2-CH2-基团(谷氨酸单元);■ n,+111'或11"是聚合度,其为IO-IOOO,优选50-300。 -0 or -NH place; ■ W represents a direct bond or a spacer based on 1 to 4 amino acid residues; surface independently, A represents a group -CH2- (aspartic unit) or -CH2-CH2- group group (glutamic unit); ■ n, + 111 'or 11 "is the degree of polymerization, which is IO-IOOO, preferably 50-300.
9. 如权利要求7或8所述的微颗粒,其中W基团代表单键。 Microparticles 7 or claim 8, wherein W represents a single bond group as claimed in claim.
10. 如权利要求l-6任何一项所述的微颗粒,其中一种或多种所述PO含有至少一个"基本上中性的"共聚羟基烷基谷氨酰胺(优选的烷基是乙基),其含有多个相同或互不相同的疏水性基团(HG)侧基。 10. l-6 microparticles of any one of claims, wherein the one or more PO containing at least one "substantially neutral" (preferred alkyl is acetic copolyhydroxyalkylglutamine group), which comprises a plurality of same or different from the hydrophobic groups (HG) side groups.
11. 如权利要求6-10任何一项所述的微颗粒,其中所述PO的所有或部分疏水性基团HG独立地选自包括以下基团的组:■ 含有6到30个碳原子的直链或支链烷氧基,并且其可以含有至少一个杂原子(优选O或N或S)或至少一个不饱和键,■ 含有6到30个碳原子的烷氧基,其具有一个或多个退火的碳环,并且可选地,含有至少一个不饱和键或至少一个杂原子(优选O或N或S),■ 含有7到30个碳原子的烷氧基芳基或芳氧基焼基,并且其可以含有至少一个不饱和键或至少一个杂原子(优选O或N或S)。 11. The microparticle of any one of claims 6-10 claim, wherein the PO of all or part of the hydrophobic groups HG independently selected from the group consisting of the following groups: ■ The containing from 6 to 30 carbon atoms, a straight-chain or branched alkoxy group, and which may contain at least one heteroatom (preferably O or N or S) or at least one unsaturated bond, ■ an alkoxy group having 6-30 carbon atoms, having one or more annealing a carbocyclic rings and optionally containing at least one unsaturated bond or at least one heteroatom (preferably O or N, or S), ■ an aryl group an alkoxy group or aryloxy firing 7-30 carbon atoms, group, and which may contain at least one unsaturated bond or at least one heteroatom (preferably O or N, or S).
12. 如权利要求6-10任何一项所述的微颗粒,其中所述疏水性基团HG选自包括以下类型的烷氧基的组:-0012(012-0"12)3.8_<:113、油烯基、生育酚基或胆甾醇基,并且W代表单键。 12. The microparticle of any one of claims 6-10 claim, wherein the hydrophobic group HG is selected from the group consisting of alkoxy type: -0012 (012-0 "12) 3.8_ <: 113, oleyl, tocopheryl or cholesteryl group, and W represents a single bond.
13. 如权利要求6-10任何一项所述的微颗粒,其中所述PO的n个HG基团各自独立地代表下式的单价基团:<formula>formula see original document page 5</formula>其中:-RS代表甲基、异丙基、异丁基、仲丁基、苄基; -RM戈表含有6到30个碳原子的疏水性基团;-l为0到6。 13. The microparticle of any one of claims 6-10 claim, wherein the n HG of the PO each independently represent a group of monovalent radical of the formula: <formula> formula see original document page 5 </ formula > wherein: -RS represents methyl, isopropyl, isobutyl, sec-butyl, benzyl; -RM Ge table containing a hydrophobic group having 6 to 30 carbon atoms; -l is 0-6.
14.如权利要求13所述的微颗粒,其中所述PO的所有或部分疏水性基团RS独立地选自包括以下基团的组:■ 含有6到30个碳原子的直链或支链垸氧基,并且其可以含有至少一个杂原子(优选O或N或S)或至少一个不饱和键,■ 含有6到30个碳原子的烷氧基,其具有一个或多个退火的碳环,并且可选地,含有至少一个不饱和键或至少一个杂原子(优选O或N或S),■ 含有7到30个碳原子的垸氧基芳基或芳氧基垸基,其可以含有至少一个不饱和键或至少一个杂原子(优选O或N或S)。 14. The microparticle according to claim 13, all or part of a hydrophobic group wherein the RS PO is independently selected from the group consisting of the following groups: ■ The straight-chain or branched-chain containing from 6 to 30 carbon atoms, embankment group, and which may contain at least one heteroatom (preferably O or N or S) or at least one unsaturated bond, ■ an alkoxy group having 6-30 carbon atoms, having one or more ring carbon annealed , and optionally containing at least one unsaturated bond or at least one heteroatom (preferably O or N, or S), ■ embankment containing group or aryloxy group, an aryl alkyl with 7 to 30 carbon atoms, which may contain at least one unsaturated bond or at least one heteroatom (preferably O or N, or S).
15. 如权利要求B或14所述的微颗粒,其中所述PO的接枝物的疏水性基团R6 选自包括以下类型的垸氧基的组:-OCH^CHrCH^^CH^油烯基、生育酚基或胆甾醇基。 15. The microparticles or B according to claim 14, wherein said hydrophobic group grafts PO R6 is selected from the following types of embankment group comprising a group: -OCH ^ CHrCH ^^ CH ^ oleyl group, tocopheryl or cholesteryl.
16. 如权利要求6-9和ll-15任何一项所述的微颗粒,其中所述聚氨基酸的主链是aL-谷氨酸根或aL-谷氨酸均聚物。 16. 6-9 ll-15 and any one of the microparticles as claimed in claim, wherein said polyamino acid backbone is aL- aL- glutamate or glutamic acid homopolymer.
17. 如权利要求6-9和ll-15任何一项所述的微颗粒,其中所述聚氨基酸的主链是aL-天冬氨酸根或aL-天冬氨酸均聚物。 17. 6-9 ll-15 and any one of the microparticles as claimed in claim, wherein said polyamino acid backbone is aL- aL- aspartate or aspartic acid homopolymer.
18. 如权利要求6-9和ll-15任何一项所述的微颗粒,其中所述聚氨基酸的主链是aL-天冬氨酸根/aL-谷氨酸根或aL-天冬氨酸/aL-谷氨酸共聚物。 18. 6-9 ll-15 and any one of the microparticles as claimed in claim, wherein said polyamino acid backbone is aL-aspartate / glutamate or aL-aL-aspartic acid / aL- acid copolymer.
19. 如权利要求6-18任何一项所述的微颗粒,其中PO含有HG,式(I)中的(n/n+m),其水平至少为10 mol%,优选至少为15moP/。 19. The microparticle of any one of claims 6-18 claim, wherein the PO containing HG, of formula (I) (n / n + m), which level of at least 10 mol%, preferably at least 15moP /. .
20. 如权利要求6-19任何一项所述的微颗粒,其中所述? 20. The microparticle of any one of claims 6-19 claim, wherein said? 0的摩尔质量在2000-100000g/mol,优选5000-40000g/mo1 。 0 molar mass in 2000-100000g / mol, preferably 5000-40000g / mo1.
21. —种制备含有至少一种活性成分(AP)的PO微颗粒的方法,这些微颗粒尤其是权利要求l-20所述的微颗粒,i. 该聚合物PO•是带有疏水性基团(HG)和亲水性基团[优选至少部分电离的可电离基团(IG)]的水溶性可生物降解的两性分子(共)聚合物,•在pHK7.0,等渗条件下,在水中自发地形成纳米颗粒的胶体悬,•并且和所述AP非共价地结合;ii. 所述微颗粒以T1试验测得的尺寸为0.5-100微米,优选l-70微米,更优选2-40 微米,该方法主要包括雾化含有AP的PO的溶液或胶体悬浮液。 21. - A method of preparing microparticles PO species contain at least one active ingredient (AP) of the microparticles, especially microparticles l 20-a, i of the polymer as claimed in claim PO • having a hydrophobic group a water-soluble group (HG) and the hydrophilic group [preferably at least partially ionized ionizable group (the IG)] biodegradable amphiphilic molecules (co) polymers, • at pHK7.0, isotonic condition, colloidal nanoparticles are formed spontaneously in water suspension, and the AP and • non-covalently bound;. ii T1 to test the microparticles dimension measured from 0.5 to 100 microns, preferably l-70 [mu] m, more preferably 2-40 microns, the method includes atomizing a solution containing an AP PO or colloidal suspension.
22. 如权利要求21所述的方法,其中存在于所述溶液或胶体悬浮液中的所述PO至少部分是纳米颗粒形式,其以T1试验测定的尺寸小于500 nm,优选为10-300 nm,并且更优选为10-100nm。 22. The method according to claim 21, wherein the solution or is present in the colloidal suspension of PO nanoparticles at least partly in the form, the size of which is measured in T1 trial less than 500 nm, preferably 10-300 nm , and more preferably 10-100nm.
23. 如权利要求21或22所述的方法,其中和至少一种活性成分(AP)相结合的PO聚合物的微颗粒分散在基本上水性的液体介质中,所述介质优选包括分散措施Mp并且冻干所得的分散体。 23. The method of claim 21 or claim 22, and wherein the at least one active ingredient (AP) in combination PO polymer microparticles dispersed in a substantially aqueous liquid medium, the dispersion medium preferably comprises measures Mp and the resulting dispersion was lyophilized.
24. 如权利要求23所述的方法,其中所述分散措施M!选自以下组:i. 和所述PO聚合物的可电离基团极性相反的多价离子,并且其包含在连续的水性相中;ii. 加入到待雾化的所述PO悬浮液/溶液中并且因此包含在雾化后的PO/AP微颗粒中的至少一种亲水性化合物(优选能用于可注射的制剂);iii. 用至少一种亲水性化合物(优选能用于可注射的制剂)包至少一层膜的至少一种微颗粒;iv. 改变pH;v. 以及措施(iMiv)中至少两种的结合; 其中措施(i)特别优选。 24. The method according to claim 23, wherein said dispersion measure M is selected from the group:!. I ionizable group and said polar polymer may PO multivalent ions of the opposite, and which comprises continuous the aqueous phase;. ii be added to the atomized PO suspension / solution and thus contained in the atomized PO / AP microparticles of at least one hydrophilic compound (preferably can be used for injectable formulation); III includes at least one film with at least one hydrophilic compound (preferably injectable formulations can be used) at least one microparticle;.. iv changing pH;. v measures and (iMiv) in at least two binding thereof; in particular wherein steps (i) preferably.
25. —种液体药物制剂,用于AP的延长释放,其含有低粘度的胶体悬浮液,所述胶体悬浮液基于含有至少一种AP的PO微颗粒,这些微颗粒是权利要求l-20任何一项所述的微颗粒或者通过权利要求21-24所述的方法获得的微颗粒。 25. - liquids pharmaceutical formulation for extended release of the AP, which contains a low viscosity colloidal suspension, a colloidal suspension based on PO microparticles comprising at least one AP, the microparticles are any of claims l-20 microparticles or by the method of any one of claims 21-24 obtained microparticles.
26. 如权利要求25所述的制剂,其含有措施M2,用于分散和至少一种AP相结合的PO微颗粒。 26. The formulation according to claim 25, which contains a measure M2, the micro-particles are dispersed for PO and at least one AP in combination.
27. 如权利要求25或26所述的制剂,其中所述悬浮液的连续相基本上是水性的。 25 or 26 formulation as claimed in claim 27, wherein the continuous phase of the suspension is substantially aqueous.
28. 如权利要求25或26所述的制剂,其中所述悬浮液的连续相基本上是有机的和水互溶的相。 25 or 26 formulation as claimed in claim 28., wherein the continuous phase of the suspension is substantially water immiscible organic phase.
29. 如权利要求25或26所述的制剂,其中所述悬浮液的连续相基本上是有机的和水不互溶的相。 29. The formulation of claim 25 or claim 26, wherein the continuous phase of the suspension and the substantially water-immiscible organic phase.
30. 如权利要求25、 26或27所述的制剂,其中所述分散措施M2选自以下组:i. 和所述PO聚合物的可电离基团极性相反的多价离子,并且其包含在连续的水性相中;ii. 加入到待雾化的所述PO悬浮液/溶液中并且因此包含在雾化后的PO/AP微颗粒中的至少一种亲水性化合物(优选能用于可注射的制剂);iii. 用至少一种亲水性化合物(优选能用于可注射的制剂)包至少一层膜的至少一种微颗粒;iv. 改变pH;V. 措施(i)-(iv)中至少两种的结合; 其中措施(i)特别优选。 30. The formulation of 25, 26 or claim 27, wherein said dispersion measure M2 is selected from the group: i ionizable groups and the polar polymer may PO multivalent ions of the opposite, and which comprises in a continuous aqueous phase;. ii PO is added to the suspension / solution to be atomized and is thus contained in the PO / the AP microparticles atomized at least one hydrophilic compound (preferably can be used injectable preparations); at least one packet III microparticles of at least one film with at least one hydrophilic compound (preferably injectable formulations can be used); IV changing pH; V steps (i) -... (iv) in connection with at least two; wherein special measures (i) is preferred.
31. 如权利要求30所述的制剂,其中所述亲水性包膜化合物选自包括以下的组:— 氨基酸类;— 聚二醇类,优选聚乙二醇类;— 共聚二醇类,优选(Poloxamer或Pluronic或Lutrol型的)乙二醇/丙二醇共聚物;4 纤维素聚合物类及其衍生物,优选羧基烷基纤维素或烷基纤维素;— 氢化或非氢化的糖类,例如海藻糖、山梨醇、甘露醇或蔗糖;— 多元醇类,如丙二醇或甘油; ~> 明胶类,优选水解明胶;— 含氮(共)聚合物类,优选自包括聚丙烯酰胺、聚-N-乙烯酰胺、聚乙烯吡咯垸酮(PVP)和聚-N-乙烯内酰胺的组;4 聚(乙烯醇)类(PVA);— 聚(谷氨酸钠); _> 及其混合物;所述亲水性包膜化合物优选含有至少一种亲水性聚合物。 Amino acids; - - polyethylene glycols, preferably polyethylene glycols; - co glycols: 31. A formulation as claimed in claim 30, wherein the hydrophilic compound selected from the group comprising envelope of the group preferably (Pluronic or Poloxamer or Lutrol type) of ethylene glycol / propylene glycol copolymers; 4 cellulose polymers and derivatives thereof, preferably cellulose or carboxyalkyl alkylcellulose; - hydrogenated or non-hydrogenated saccharides, such as trehalose, sorbitol, mannitol or sucrose; - polyols, such as propylene glycol or glycerol; ~> gelatin, hydrolyzed gelatin preferred; - a nitrogen-containing (co) polymers, preferably from the group consisting of polyacrylamide, poly - N- vinyl amide, ketone embankment polyvinylpyrrolidone (PVP) and poly-lactam -N- ethylene group; 4 poly (vinyl alcohol) (PVA); - poly (sodium glutamate); _> and mixtures thereof; said envelope hydrophilic compound preferably contains at least one hydrophilic polymer.
32. 如权利要求25和27或28所述的制剂,其中所述分散措施含有一种存在于和水互溶或和水不互溶的连续相中的亲脂性液体,其熔点优选低于或等于15'C。 25 and 32. The formulation of claim 27 or claim 28, wherein said dispersion contains measures lipophilic liquids and be present in one kind or water-miscible and water-immiscible continuous phase, preferably having a melting point lower than or equal to 15 'C.
33. 如权利要求32所述的制剂,其中所述亲脂性液体含有至少一种饱和脂肪酸的甘油三酯的混合物或者至少一种植物油或至少一种脂质或至少一种脂质衍生物或至少一种脂肪酸或至少一种脂肪酸衍生物。 33. The formulation according to claim 32, wherein the lipophilic liquid mixture containing at least one saturated fatty acid triglyceride or at least one vegetable oil or at least one lipid or lipid derivative or at least one at least one fatty acid or at least one fatty acid derivative.
34. 如权利要求33所述的制剂,其中所述亲脂性液体含有• 至少一种源自椰子油的饱和CVdo脂肪酸的甘油三酯的混合物;• 至少一种植物油,优选大豆油、棕榈油、亚麻籽油、棉籽油、芝麻油、葵花籽油或花生油;• 至少一种脂质,优选脂质卵磷脂、合成或天然的维生素E;• 至少一种脂质衍生物,优选花生四烯酰磷脂酰胆碱和硬脂酰磷脂酰胆碱;• 至少一种脂肪酸,优诜袖酴、囪W蔻酴、棕飽酴、硬脂酴及其盐类;• 至少一种脂肪酸衍生物,优选单-或三-甘油酯衍生物、油酸乙酯、乳酸月桂酯、硬脂酸甘油酯、棕榈酸脱水山梨聚醇酯、硬脂酸脱水山梨醇酯、脱水山梨醇一油酸酯或聚山梨醇酯;• 及其混合物;其条件是当上面列出的某些产品如果在例如低于或等于15C时单独使用时不是液体形式,那么这些产品可以和其它产品混合,以使其在例如低于或 34. The formulation according to claim 33, wherein the lipophilic • at least one liquid mixture of triglycerides containing saturated fatty acid CVdo derived from coconut oil; • at least one vegetable oil, preferably soybean oil, palm oil, linseed oil, cottonseed oil, sesame oil, sunflower oil, or peanut oil; • at least one lipid, preferably phosphatidylcholine lipids, synthetic or natural vitamin E; • at least one lipid derivative, preferably phosphatidyl arachidonic stearoyl phosphatidyl choline and choline; • at least one fatty acid, preferably Shen sleeve yeast, yeast chimney W coronene, full brown yeast, yeast stearyl and salts thereof; • at least one fatty acid derivative, preferably mono- - or tri - glyceride derivatives, ethyl oleate, lauryl lactate, glyceryl stearate, palmitic acid, polyethylene sorbitan esters, sorbitan monostearate, sorbitan monooleate or polysorbate alcohol esters; • and mixtures thereof; with the proviso that when some of the products listed above, for example, if used alone at 15C is not less than or equal to a liquid form, then these products can be mixed with other products, so as for example low in or 等于15'C 时是液体形式。 Equal to 15'C liquid form.
35. 如权利要求25和27或28以及可选地32、 33或34所述的制剂,其中所述分散措施包括用至少一种膜形成包膜化合物(优选其能用于可注射制剂)对所述微颗粒进行包膜。 35. The formulation of claim 25 and 27 or 28 and optionally 32, 33 or 34 pairs, wherein said dispersion include forming compound at least one membrane envelope (which can be preferably used in an injectable formulation) the microparticles be coated.
36. 如权利要求35所述的制剂,其中所述膜形成包膜化合物含有至少一种疏水性聚合物,选自包括聚交酯、聚羟基乙酸、聚(交酯-共-乙交酯)、聚原酸酯、聚酸酐、聚(羟基丁酸)、聚己酸内酯、聚(碳酸垸基酯)、水不溶性PO聚合物、它们的衍生物和混合物的组。 36. The formulation according to claim 35, wherein the envelope film is formed of a compound containing at least one hydrophobic polymer selected from the group comprising polylactide, polyglycolic acid, poly (lactide - co - glycolide) , polyorthoesters, polyanhydrides, poly (hydroxybutyric acid), polycaprolactone, poly (ester carbonate embankment), water-insoluble polymer PO, the group of derivatives and mixtures thereof.
37. 如权利要求25和27或29以及可选地权利要求31-35中至少一项所述的制剂,其中所述膜形成包膜化合物是脂质性质的,其熔点优选高于或等于15i:,并且含有至少一种饱和脂肪酸的甘油三酯的混合物或者至少一种植物油或至少一种脂质或至少一种脂质衍生物或至少一种脂肪酸或至少一种脂肪酸衍生物或者这些产品的混合物。 37. Claim 25 and 27 or 29 and optionally a formulation as claimed in claims 31-35 to at least one of the claims, wherein said film-forming compound is a lipid envelope nature, preferably having a melting point higher than or equal to 15i :, and a mixture comprising at least one triglyceride of saturated fatty acids, or at least one vegetable oil or at least one lipid or lipid derivative, or at least one of the at least one fatty acid or at least one fatty acid or a derivative of these products mixture.
38. —种重构药盒,尤其用于重构如权利要求24-37任何一项所述的制剂,其包括:• 含有至少一种AP的PO微颗粒,这些微颗粒是权利要求l-20任何一项所述的微颗粒或者通过权利要求21或22所述的方法获得的微颗粒;• 和一种重构液体,选自包括以下的组:— 基本上水性的液体; 基本上有机的和水互溶的液体;— 以及基本上有机的和水不互溶的液体。 38. - species reconstruction kit, in particular for the formulation of any one of claims 24-37 reconstructed, which include: • contain at least one AP-PO microparticles, the microparticles are according to claim l- 20 any microparticles or microparticles method according to claim 21 or 22 obtained; • reconstitution liquid and one selected from the group comprising: - a substantially aqueous liquid; substantially organic and a water-miscible liquid; - and a substantially water immiscible organic liquid.
39. —种重构方法,尤其用于重构如权利要求24-37任何一项所述的制剂,其主要包括:• 混合^含有至少一种AP的PO微颗粒,这些微颗粒是权利要求l-20任何一项所述的微颗粒或者通过权利要求21或22所述的方法获得的微颗粒;^和一种重构液体,选自包括以下的组:— 基本上水性的液体;— 基本上有机的和水互溶的液体;— 以及基本上有机的和水不互溶的液体;• 并且搅拌该混合物。 39. - species reconstruction method for reconstructing particular formulation as claimed in any one of claims 24-37, which mainly include: • ^ PO microparticles comprising mixing at least one AP, the microparticles are according to claim l-20 fine particles of any one of the microparticles or by the method as claimed in claim 21 or 22 obtained; ^ reconstitution liquid and one selected from the group comprising: - a substantially aqueous liquid; - and substantially water immiscible organic liquid; - and an organic liquid substantially immiscible in water; • and the mixture was stirred.
40. —种固体药物制剂,用于AP的延长释放,其含有干燥粉末形式:• 其基于含有至少一种AP的PO微颗粒,这些微颗粒是权利要求l-20任何一项所述的微颗粒或者通过权利要求21或22所述的方法获得的微颗粒;• 或者由权利要求24-37任何一项所述的制剂获得。 40. - one solid pharmaceutical formulation for extended release of the AP, which contains a dry powder forms: • A PO based microparticles comprising at least one AP, the microparticles are micro-l-20 according to any one of claims particles or fine particles by the method as claimed in claim 21 or 22 obtainable; • formulation of any one of claims 24-37 or claims to be obtained.
41. 如权利要求40所述的固体药物制剂,用于吸入或者肺部给药。 41. The solid pharmaceutical formulation according to claim 40, for inhalation or pulmonary administration.
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