CN101756961A

CN101756961A - Drug formulations having long and medium chain triglycerides

Info

Publication number: CN101756961A
Application number: CN200910176277A
Authority: CN
Inventors: 埃德加·H·厄尔马; 罗伯特·曼斯菲尔德; 马库斯·贝姆
Original assignee: Conforma Therapeutics Corp
Current assignee: Conforma Therapeutics Corp
Priority date: 2003-03-13
Filing date: 2003-10-04
Publication date: 2010-06-30
Also published as: AU2003277299A1; EP1605931A4; WO2004082676A1; JP2006514994A; EP1605931A1; AU2003277299B2; CA2518836A1

Abstract

Drug formulations having emulsifying agents and both medium and long chain triglycerides are described. In preferred embodiments, the long chain triglycerides negate or lessen deleterious central nervous system effects that are caused by medium chain triglycerides.

Description

Contain the pharmaceutical preparation of long-chain and medium chain triglyceride

The application is the dividing an application that be on October 4th, 2003, denomination of invention the applying date for the application for a patent for invention of " containing the pharmaceutical preparation of long-chain and medium chain triglyceride ".

Related application

The application requires following priority and quotes its full content in the lump as a reference at this: the title that Ulm etc. submitted on July 29th, 2003 is No. 60/491,050, the U.S. Provisional Patent Application of " Ansamycin preparation and preparation thereof and using method "; The title that Ulm etc. submitted on June 12nd, 2003 is No. 60/478,430, the U.S. Provisional Patent Application of " based on preparation and the preparation and the using method of phospholipid "; The title that Ulm etc. submitted on March 13rd, 2003 is No. 60/454,812, the U.S. Provisional Patent Application of " HSP90 inhibitor formulations and data "; The title of on April 4th, 2003 submitting to Ulm etc. be that the PCT of " novel Ansamycin preparation and preparation and using method " applies for PCT/US03/10533 number, it requires the priority of the U.S. Provisional Patent Application 60/371,668 of the same title submitted on April 10th, 2002.

Technical field

The present invention relates generally to pharmaceutical preparation and method, and relate to the emulsification preparation of Ansamycin In a more specific embodiment, as 17-AAG.

Background technology

Comprise having in below describing and help understand information of the present invention.This is not that approval is a prior art or relevant with the present invention of applying in this any information that provides, and any public publication perhaps clear and definite or that hint is mentioned is a prior art.

17-allyl amino-geldanamycin (17-AAG) is a kind of synthetic analogues of geldanamycin (GDM).These two kinds of molecules all belong to a big class antibiotic molecule that is called as Ansamycin.GDM at first separates from microorganism streptomyces hygroscopicus (Streptomyceshygroscopicus), being confirmed as at first is some kinase whose effective inhibitor, the effect that showed them afterwards is by promoting the kinases degraded, particularly targeting " molecular chaperone ", for example heat shock protein 90 (HSP90).Afterwards, various other Ansamycins also demonstrated this activity more or less, and wherein 17-AAG is one of most promising, the deep clinical research of just implementing by National Cancer Institute (NCI) at present with it as object of study.Referring to, for example, Federal Register, 66 (129): 35443-35444; Erlichmanet al., Proc.AACR (2001), 42, abstract 4474.

HSP90 is ubiquitous chaperone protein, and it folds, activates and assemble relevant with numerous protein, and these protein comprise the key protein matter that relates to control of signal transduction, cell cycle and transcriptional regulatory.Personnel report according to the study, and the HSP90 chaperone protein is relevant with important signal-proteins, for example steroid hormone receptor and protein kinase, comprise as, Raf-1, EGFR, v-Src family kinase, Cdk4 and ErbB-2 (Buchner J., 1999, TIBS, 24:136-141; Stepanova, L.et al., 1996, Genes Dev.10:1491-502; Dai, K.et al., 1996, J.Biol.Chem.271:22030-4).Research further shows some auxilliary chaperone, as, Hsp70, p60/Hop/Sti1, Hip, Bag1, HSP40/Hdj2/Hsj1, immunophilin, p23 and p50, can help the HSP90 function performance (referring to, for example, Caplan, A., Trends in Cell Biol., 9:262-68 (1999)).

It is believed that the ansamycins antibiotic, for example Antibiotic TAN 420F (HA), geldanamycin (GM) and 17-AAG, can bring into play its anticancer effect (Stebbins by combining closely with the terminal ATP binding pocket of N-(ATP-binding pocket) of HSP90, C.et al., 1997, Cell, 89:239-250).This bag is high conservative, and and the ATP binding site of dna gyrase between have weak homology (Stebbins, C.et al., the same; Grenert, J.P.et al., 1997, J.Biol.Chem., 272:23843-50).In addition, there has been the result to show that ATP and ADP all combine with this bag with low-affinity, and had weak atpase activity (Proromou, C.et al., 1997, Cell, 90:65-75; Panaretou, B.et al., 1998, EMBO J., 17:4829-36).Studies show that in vitro and in vivo, the terminal bag of N-is occupied the function that has changed HSP90 and has suppressed Protein Folding by Ansamycin and other HSP90 inhibitor.Under high concentration, Ansamycin and other HSP90 inhibitor can stop (Scheibel, T.H.et al., 1999, the Proc.Natl.Acad.Sci.U S A 96:1297-302 of combining of protein substrate and HSP90; Schulte, T.W.et al., 1995, J.Biol.Chem.270:24585-8; Whitesell, L., et al., 1994, Proc.Natl.Acad.Sci.U S A 91:8324-8328).Shown also simultaneously that Ansamycin can suppress release (Schneider, C.L.et al., 1996, Proc.Natl.Acad.Sci.USA, the 93:14536-41 of the ATP dependence of conpanion's associated protein substrate; Sepp-Lorenzino et al., 1995, J.Biol.Chem.270:16580-16587).Which kind of situation no matter, and a kind of process degraded that relies on ubiquitin that substrate is all existed in the proteasome (Schneider, C., L., the same; Sepp-Lorenzino, L., et al., 1995, J.Biol.Chem.270:16580-16587; Whitesell, L.et al., 1994, Proc.Natl.Acad.Sci.USA, 91:8324-8328).

The stabilization removal of this substrate occurs in tumor and the non-transformed cell, show simultaneously a subclass Signal Regulation thing effective especially, as Raf (Schulte, T.W.et al., 1997, Biochem.Biophys.Res.Commun.239:655-9; Schulte, T.W., et al., 1995, J.Biol.Chem.270:24585-8), nuclear steroid receptor (Segnitz, B., andU.Gehring.1997, J.Biol.Chem.272:18694-18701; Smith, D.F.et al., 1995, Mol.Cell.Biol.15:6804-12), v-src (Whitesell, L., et al., 1994, Proc.Natl.Acad.Sci.U S A 91:8324-8328) and some stride film tyrosine kinase (Sepp-Lorenzino, L.et al., 1995, J.Biol.Chem.270:16580-16587), for example EGF receptor (EGFR) and Her2/Neu (Hartmann, F., et al., 1997, Int.JCancer 70:221-9; Miller, P.et al., 1994, Cancer Res.54:2724-2730; Mimnaugh, E.G., et al., 1996, J.Biol.Chem.271:22796-801; Schnur, R.et al., 1995, J.Med.Chem.38:3806-3812), CDK4 and mutant p53.Erlichman?et?al.，Proc.AACR(2001)，42，abstract?4474。Inductive these the proteinic losses of Ansamycin cause some adjusting path to suffer selective destruction, and the moment growth retardation (Muise-Heimericks that causes at cell cycle, R.C.et al., 1998, J.Biol.Chem.273:29864-72), and the apoptosis of handled cell and/or differentiation (Vasilevskaya, A.et al., 1999, Cancer Res., 59:3935-40).

Recently, the WO 01/72779 (PCT/US01/09512) of Nicchitta etc. shows, it is contemplated that HSP90 has a kind of different conformation in heat shock and/or after by fluorogen bis-ANS combination.Particularly, Nicchitta etc. have proved, inductive this conformation is compared with the different shape of dominant HSP90 in normal cell, and having shown has higher affinity to some HSP90 part.The application PCT/US02/39993 that owns together has further supported this discovery by proof cancerous cell lysate as the purposes and the application in the good source of high-affinity HSP90.

Except anticancer and antitumorigenic activity, the HSP90 inhibitor also relates to multiple other application, comprise as the sick medicine of anti-inflammatory agent, infection, treat the medicine of autoimmune medicine, treatment apoplexy, ischemia, cardiac disorder and help lend some impetus to neuranagenesis medicine (referring to, for example, Rosen et al., WO 02/09696 (PCT/US01/23640); Degranco et al., WO 99/51223 (PCT/US99/07242); Gold, United States Patent (USP) 6,210,974B1; DeFranco et al., United States Patent (USP) 6,174,875).Overlapping to some extent with some top content, document has reported that also it also can treat fiber generative nature disease, includes but not limited to scleroderma, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, liver cirrhosis, keloid formation, interstitial nephritis and pulmonary fibrosis.(Strehlow，WO02/02123；PCT/US01/20578)。In addition, the modulation of HSP90, instrumentality and application are also reported in following document: PCT/US03/04283, PCT/US02/35938, PCT/US02/16287, PCT/US02/06518, PCT/US98/09805, PCT/US00/09512, PCT/US01/09512, PCT/US01/23640, PCT/US01/46303, PCT/US01/46304, PCT/US02/06518, PCT/US02/29715, PCT/US02/35069, PCT/US02/35938, PCT/US02/39993,60/293,246,60/371,668,60/331,893,60/335,391,06/128,593,60/337,919,60/340,762 and 60/359,484.

At present, the same with other many lipophilic drugs, Ansamycin is difficult to be prepared into pharmaceutical formulation, especially the used for intravenous injection preparation.Up to now, attempted it is prepared into lipid vesicle and oil-in-water emulsion, but this all needs complicated procedure of processing, coarse or clinical unacceptable solvent and/or causes the preparation instability.Usually can be referring to Vemuri, S.andRhodes, C.T., Preparation and characterization of liposomes astherapeutic delivery systems:a review, Pharmaceutica Acta Helvetiae 70, pp.95-111 (1995); Also can be referring to announcing that publication number is the PCT/US99/30631 of WO00/37050 on June 29th, 2000.The application requires the application PCT/US03/10533 that owns together of priority to describe the Emulsion that contains medium chain triglyceride.But the medium-chain fatty acid and the triglyceride that carry them may cause metabolism to generate caprylate, thereby cause undesirable central nervous system's effect, for example, and the change of drowsiness, nauseating, sleepy and EEG.Referring to Cotter et al., Am.J.Clin.Nutr.50:794-800 (1989); Miles etal., Journal of Parenteral and Enteral Nutrition 15:37-41 (1991); Traul et al., Food Chem.Toxicol.38:79-98 (2000).Up to now, this class side effect is by using long-chain fatty acid partly to improve with the form of supplementary, and its mechanism of action is that long-chain fatty acid is competed crucial caprylate by way of enzyme with higher joint efficiency.But, according to the applicant up to now known to, they are not and medium chain triglyceride and Ansamycin use in conjunction.

Therefore, need other preparation and preparation method thereof, it can create relatively simply and can improve the above-mentioned shortcoming that one or several is caused by medium chain triglyceride usually.

Summary of the invention

By long chain triglyceride is formulated together as the component and the chemical compound of preparation, find the applicant this claimed preparation can be used in toleration better intravenous use for example Ansamycin of lipophilic compound.

In first aspect, the invention is characterized in a kind of pharmaceutical composition, it comprises pharmaceutically active compound, and Ansamycin for example as 17-AAG, closes use with a kind of emulsifying agent (for example phospholipid, as the phospholipid of being found) and line of oils in lecithin.Oil can and preferably comprise long chain triglyceride.Said composition also can comprise medium chain triglyceride.Emulsifying agent constitutes fat mutually with oil is common.

In certain embodiments, fat accounts for the 5-30%w/w of total preparation mutually, preferred 5-20%w/w.

In certain embodiments, total w/w percentage ratio of long chain triglyceride is no more than 10%, and more preferably scope is 7% or lower, and more preferably scope is 6% or lower, to adapt to viscosity limitation.

In certain embodiments, medium chain triglyceride is 10: 0.0001 to 0.0001: 10 with the w/w ratio of long chain triglyceride, more preferably 10: 1 to 1: 10.

In certain embodiments, phospholipid accounts for the 3-10%w/w of gross weight.

In certain embodiments, triglyceride accounts for the 5-20%w/w of gross weight.

In certain embodiments, triglyceride is, or to small part be the form of the oil that exists of nature, vegetable oil for example is as soybean oil, Oleum sesami, safflower oil and Semen Maydis oil.

In certain embodiments, said composition further comprises one or more in water, antiseptic (for example sodium ethylene diamine tetracetate), antifreeze, buffer, chelating agen and the tonicity contributor (tonicifier).

In certain embodiments, medicine is 17-AAG (17-allyl amino geldanamycin mycin, a 17-allyl amino-17-demethoxylation-geldanamycin), and amount is 0.5mg/ml-4mg/ml or the 0.05%w/w-0.4%w/w that accounts for total weight of formulation.

In one embodiment, said composition contains the 17-AAG of following composition: 2mg/ml, 3.3% soybean oil, 6.6% lecithin, 9.9% Miglyol 812N, 7.15% sucrose, and water.

In another embodiment, said composition contains the 17-AAG of following composition: 2mg/ml, 7.5% lecithin, 15% Miglyol 812N, 10% sucrose, and water.

17-AAG is a 17-allyl amino geldanamycin mycin, has following structure:

In certain embodiments, compositions of the present invention also comprises short chain triglyceride.

In an especially preferred embodiment, said composition comprises the long chain triglyceride of q.s to reduce or to avoid the generation of central nervous system (CNS) effect of medium chain triglyceride mediation, particularly also comprises in said composition under the situation of medium chain triglyceride.The CNS effect normally negative, do not wish the effect that takes place, include but not limited to drowsiness, nauseating, sleepy and EEG one or more in changing.Yet in certain embodiments, this (a bit) acts under the given background and may wish, so that medium chain triglyceride is with respect to the amount increase of long chain triglyceride.

In certain embodiments, said composition is refrigerated and/or freeze dried, as summarizing among the PCT/US03/10533.In freeze dried embodiment, the percentage by weight of triglyceride, phospholipid, medicine and each component of nonvolatile element all must increase and also surpasses above-mentioned scope, to meet lost the water that exists at first but lose and other volatile components after lyophilizing after their relative fractional increase.

In certain embodiments, said composition also can be prepared in inert gas conditions and/or be stored, and for example is stored in dark and/or lighttight bottle, phial or the ampoule.

Also relate to the combination of any above-mentioned embodiment in the time of suitable.

On the other hand, the invention is characterized in the method for a kind of patient's of minimizing generation by central nervous system's effect of medium chain triglyceride mediation, comprise: a kind of pharmaceutical preparation that comprises pharmaceutical active medicine and long chain triglyceride is provided, the amount of described long chain triglyceride is enough to reduce or avoid the generation of central nervous system's effect of medium-chain fatty acid mediation, and the said goods is administered to the patient.The embodiment of this respect can be with reference to any compositions embodiment and the combination thereof of above-mentioned aspect.

On the other hand, the invention is characterized in that the method that makes pharmaceutical composition, preparation and the product by biology being used pharmacy effective dose are with treatment or pre-anti-biological said method and product as mammalian diseases.Described disease, is preferably selected from diseases such as ischemia, proliferative disease and nerve injury, and comprises the HSP90 inhibitor during mammiferous situation in treatment at least, as one or more Ansamycins as the pharmaceutical active medicine.Proliferative disease includes but not limited to tumor and cancer, inflammatory diseases, fungal infection, yeast infection and viral infection.In some preferred embodiment, mammal is the people.In some preferred embodiment, administering mode is intravenous injection, and even so, in the more detailed description, other administering modes also are admissible below.

According to specific embodiment, advantage of the present invention comprises following one or multiple advantages: be easy to preparation, use that clinical acceptable agents (as reducing the toxicity to environment and/or patient), stability of formulation improve, are easy to load and transport and put in storage, prescription and bedside is simple to operate, intravenous and whole body tolerance during administration and having avoided often caused by intravital medium-chain fatty acid and triglyceride some undesirable side effects.Other advantage, aspect and embodiment will be conspicuous by following accompanying drawing, detailed description and claim.

Description of drawings

Fig. 1 shows that the drowsiness of rat alleviates owing to add long-chain fatty acid in comprising the preparation of medium chain triglyceride.

The explanation of preferred embodiment

Preparation of the present invention has special advantage on being suitable for the water soluble drug of patient with intravenous injection and otherwise administration.Compound method is simple relatively, adopts clinical acceptable agents usually, and the product of preparation all has the advantage that surpasses existing method and product on storage, stability and biological tolerance.

Definition

Below term have following implication, and the following term that does not particularly point out also has this area and is used to the implication that has usually:

Term " pharmaceutically active compound " has identical meanings with " medicine ", is meant in vivo or the external chemical compound that is applied to cultured cells or any direct or indirect performance biological effect of biochron of working as.Medicine preferably can be packaged in the liposome and/or emulsifying, though and be not essential, this medicine is normally lipophilic.

Term " evaporation " and " lyophilizing " also do not mean that must 100% to remove and desolvate and solution, may just remove less ratio.But, in freeze dried embodiment, preferably remove basically, preferably remove about 95% or more.

" inert gas conditions " is meant to compare with the gas in the standard atmosphere condition to have relatively low reactive gas condition.An example of this inert gas conditions is to use pure or pure basically nitrogen in process for preparation.Those skilled in the art also knows for other situation.

Term " hydration " or " rehydrated " are meant the adding aqueous solution, as the buffer of water or physical compatibility, and for example hanks' solution, Ringer's solution or normal saline buffer solution.

Term " about " is meant the scope that contains about described value 20% deviation.Term " comprise " and " between " or during " between approximately " logotype, mean the endpoint value that comprises described scope.

Term " Ansamycin " is a generalized term, characterizes the chemical compound with " loop " structure, and described " loop " structure comprises one of benzoquinone, benzo hydroquinone, naphthoquinone or naphtho-hydroquinone part by the long-chain bridge joint.The example of naphthoquinone or naphtho-hydroquinone compound is respectively important clinically medicament benemicin and rifamycin.The example of benzoquinone compound is that geldanamycin (comprises its synthesis of derivatives, 17-allyl amino-17-demethoxylation geldanamycin (17-AAG) and 17-N, N-dimethylamino-ethylamino-17-demethoxylation geldanamycin (DMAG)), dihydro geldanamycin and herbimycin.The example of benzo hydroquinones is Mike's rhzomorph.According to Ansamycin of the present invention and benzoquinone Ansamycin can be synthetic, natural generation or the combination of the two, i.e. " semisynthetic ", and can comprise the form of dimer and coupling variant and prodrug.Typical benzoquinone Ansamycin of useful in the present invention some and preparation method thereof includes but not limited to that as United States Patent (USP) 3,595 955 (having described the preparation of Ansamycin), 4,261,989,5,387,584 and 5,932 are described in 566.Geldanamycin also can be buied, and for example available from CN Biosciences, it is Merck KGaA,, Darmstadt, the tame affiliated company of Germany, general headquarters are positioned at San Diego, California, USA (article No. 345805).Geldanamycin derivant 4,5-dihydro geldanamycin and hydroquinone thereof the biochemistry purification from streptomyces hygroscopicus (ATCC 55256) culture is Pfizer Inc the application people, the invention people is Cullen etc., be disclosed on July 22nd, 1993, publication number is WO93/14215, and international application no is to describe in the application of PCT/US92/10189; Catalytic hydrogenation by geldanamycin synthesizes 4, and a kind of selectable method of 5-dihydro geldanamycin also is known.Referring to, for example, Progress in the Chemistry of OrganicNatural Products, Chemistry of the Ansamycin Antibiotics, 33:278 (1976).Other useful in various embodiments of the present invention Ansamycins have description in the document that above-mentioned " background technology " part is quoted.

" oil " comprises fatty acid and contains the glyceride of fatty acid, monoglyceride for example known in the art, diglyceride and triglyceride.Fatty acid of Shi Yonging and glyceride can be saturated and/or undersaturated in the present invention, and is natural and/or synthetic, charged or neutral.It can be synthetic or semi-synthetic fully " synthesizing ", and these terms all are known in the art.Oil also can be homogeneous or inhomogenous on component and/or source.

Herein, " medium chain triglyceride " is that to contain length be 8-12 straight chain carbon atom, and more preferably length is the triglyceride compositions of the fatty acid of 8-10 carbon atom.Various embodiments of the present invention comprise that (Cranford, NJ USA) provide by CONDEA in use 812.

812 roughly comprise 50-65% sad (8 carbon atoms) and 30-45% capric acid (10 carbon atoms).Also can have caproic acid (6 carbon atoms), reach as high as approximately 2%, lauric acid (12 carbon atoms) is so same.Myristic acid (14 carbon atoms) exists with amount (the highest by 1%) still less.Condea also provides

810,818,829 and 840, can predict one or more these other Solution and other medium chain triglyceride solution also can more or less be successfully used in various aspects of the present invention and the embodiment.As for the latter, those skilled in the art are familiar with its character, source and/or preparation method, and just can obtain or prepare without too many research or experiment.Miglyol 812N in European Pharmacopoeia as medium chain triglyceride, in British Pharmacopoeia as fractionated Oleum Cocois, and in Japanese Pharmacopoeia, monograph is all arranged as caprylic/capric triglyceride.Other sources of medium chain triglyceride comprise Oleum Cocois, palm-kernel oil and butter.

" short chain triglyceride " is to contain the triglyceride compositions that length is less than the fatty acid of 8 straight chain carbon atoms.

" long chain triglyceride " is to contain the triglyceride compositions of length greater than the fatty acid of 12 straight chain carbon atoms.Their common source is a vegetable oil, soybean oil for example, and it contains 55-60% linoleic acid (9, the 12-octadecadienoic acid) usually, 22% oleic acid (suitable-the 9-octadecenoic acid) and a small amount of Palmic acid and stearic acid.

Term " weak point ", " in " and " length " also can be used for independent fatty acid, in this case, its definition comprises respectively and is less than 8 straight chain carbon atoms, 8-12 straight chain carbon atom and greater than 12 straight chain carbon atoms.

" emulsifying agent " and " surfactant " synonym include but not limited to phospholipid, as lecithin." lecithin " is the naturally occurring mixture that is connected to stearic acid, Palmic acid and oleic diglyceride on the phosphocholine ester.Nomenclature surface-active agent or emulsifying agent also comprise phosphatidylcholine, and this compounds is well-known.The preferred emulsifier of using among the present invention is a soybean lecithin, as American Lecithin Company (Oxford, CT, the Phospholipon 90G that USA) provides.Phospholipon 90G has been applied in the nutrition product of parenteral introduction in the early time, as

Middle concentration is about 1.2%,

Be about 1% in (doxorubicin),

Be about 2% in (amphotericin B), and In be about 1.2%.About the latter, can be referring to for example United States Patent (USP) 6,140,374.In the amount of other components of water and/or dissolving surfactant, surfactants exists with the concentration of about 0.5-25%w/v usually.Surfactant concentration preferably is about 0.5-10%w/v, is most preferably 1-8%w/v.

Examples of anionic surfactants comprises sodium lauryl sulfate; triethanolamine lauryl sulfate ester; sodium laureth sulfate; polyoxyethylene nonylplenyl ether sodium sulfate; triethanolamine polyoxyethylene lauryl ether sulfuric ester; the cocoyl sodium sarcosinate; N-cocos nucifera oil acyl methyl taurine sodium; polyoxyethylene (Cortex cocois radicis) sodium alkylether sulphate; diether hexyl sodium sulfo-succinate; the alpha-olefin sodium sulfonate; dodecylphosphoric acid sodium; the polyoxyethylene lauryl ether sodium phosphate; perfluoroalkyl carboxylate (made by Daikin Industries Ltd., commodity are called UNIDINE DS-101 and 102).

The example of cationic surfactant comprises dialkyl group (C ₁₂-C ₂₂) alkyl dimethyl ammonium chloride, alkyl (Cortex cocois radicis) dixylyl ammonium chloride, 18-amine. acetate, tetradecy lamine acetate, tallow alkyl Propylenediamine Tetraacetylimide acetate, octadecyl trimethyl ammonium chloride, alkyl (Adeps Bovis seu Bubali) trimethyl ammonium chloride, Dodecyl trimethyl ammonium chloride, alkyl (Cortex cocois radicis) trimethyl ammonium chloride, hexadecyltrimethylammonium chloride, the biphenyl trimethyl ammonium chloride, alkyl (Adeps Bovis seu Bubali)-imidazoline quaternary salt, myristyl methylbenzyl chlorination ammonium octadecyl dixylyl ammonium chloride, two oil base alkyl dimethyl ammonium chlorides, polyoxyethylene dodecyl monomethyl ammonium chloride, polyxyethylated (C ₁₂-C ₂₂) benzyl ammonium chloride, polyoxyethylene lauryl monomethyl ammonium chloride, 1-ethoxy-2-alkyl (Adeps Bovis seu Bubali)-imidazoline quaternary salt and contain siloxane group as the silicone cationic surfactant of hydrophobic group, contain the fluorine-containing cationic surfactant (by Daikin Industries Ltd made, commodity be called UNIDINE DS-202) of fluoro-alkyl as hydrophobic group.

The example of non-ionic surface active agent comprises polyoxyethylene lauryl ether, the polyoxyethylene tridecyl ether, Polyoxyethylene cetyl ether, the polyoxyethylene polyoxypropylene cetyl ether, polyoxyethylene stearyl ether, polyoxyethylene oleyl ether, the polyoxyethylene nonylplenyl ether, NONIN HS 240, the polyoxyethylene mono laurate, the polyoxyethylene monostearate, polyoxyethylene list oleic acid, Arlacel-20, Arlacel-60, Arlacel-40, Arlacel-60, Arlacel-80, Arlacel-83, sorbitan trioleate, polyoxyethylene sorbitan monolaurate, the polyethenoxy sorbitan monopalmitate, the polyethenoxy sorbitan monostearate, polyoxyethylene sorbitan monooleate dehydration, the polyoxyethylene polyoxypropylene block polymer, polyglyceryl fatty acid ester, the silicone oil of polyether-modified is (by Toray DowCorning Silicone Co., Ltd. make, commodity are called SH3746, SH3748, SH3749 and SH3771), the perfluoroalkyl ethylene oxide adduct (is made by Daikin Industries Ltd, commodity UNIDINE DS-401 by name and DS-403), fluoro-alkyl oxireme adduct (is made by Daikin Industries Ltd, commodity are called UNIDINE DS-406), and perfluoroalkyl oligomer (made by Daikin Industries Ltd, commodity are called UNIDINE DS-451).

" physiology acceptable carrier " is meant a kind of carrier or diluent, and it can not produce significant stimulation to biology nor the biological activity of the chemical compound of using and character are disappeared.

" excipient " is meant and joins the material that can further promote compound administration in the pharmaceutical compositions.The example of excipient includes but not limited to calcium carbonate, calcium phosphate, multiple sugar and dissimilar starch, cellulose and cellulose derivative, gelatin, vegetable oil and Polyethylene Glycol.These also can be the physiology acceptable carriers, as mentioned above, and sucrose for example.Further belong in the excipient range of definition is filler." filler " generally keeps its conformation to provide machinery support to give the lyophily preparation by the substrate that makes drying agent.Be preferably sugar.Sugar used herein includes but not limited to monosaccharide, disaccharide, oligosaccharide and polysaccharide.Concrete example includes but not limited to fructose, glucose, mannose, trehalose, sorbose, xylose, maltose, lactose, sucrose, dextrose, reaches glucosan.Sugar also comprises sugar alcohol, as mannitol, sorbitol, inositol, galactitol, xylitol and 1,2,3,4,5-pentanepentol.Also can use the mixture of sugar according to the present invention.Multiple filler also can be brought into play the function of isotonic agent as glycerol, sugar, sugar alcohol and monosaccharide and disaccharide.Preferred filler is chemically inert for medicine and does not have or have extremely limited deleterious side effect or toxicity under service condition.Except that the filler carrier, other carriers also may reach or may not reach the purpose of filler, comprise, as known in the art and the adjuvant and the diluent that obtain easily.

The preferred a kind of filler of the present invention is a sucrose.Be not subjected to any theoretical constraint, think that sucrose forms unformed glassy mass when freezing and lyophilizing subsequently, thereby by in hard glassy mass, forming the possible stability that the dispersive oil droplet that comprises active component strengthens preparation.Because sugar has substituted the water that loses when lyophilizing, therefore also can enhanced stability.Be not hydrone, but glycan molecule is by the phospholipid bonding on hydrogen bond and the interface.Have these features and can include but not limited to cellulosics by substituted other filleies, for example, corn starch, wheaten starch, rice starch, potato starch, gelatin, tragakanta, methylcellulose, hydroxypropyl emthylcellulose, sodium carboxymethyl cellulose and/or polyvinylpyrrolidone (PVP).If desired, also can add disintegrating agent, as crosslinked polyvinylpyrrolidone, agar or alginic acid or its salt such as sodium alginate.

Phrase in the claim " reduce or avoid taking place amount by central nervous system's effect of medium chain triglyceride mediation " need not too many experiment and promptly can utilize starting point described herein to determine by experience by those of ordinary skill.

Emulsification

Known in this field, the Emulsion that comprises oil phase and water is as the carrier of treatment effective ingredient or as the source of parenteral absorption.Emulsion can be oil-in-water or water-in-oil type.As current situation, if therapeutic component dissolves in oil phase especially, then oil-in-water type is an embodiment preferred.Simple Emulsion is the system of thermodynamic instability, because wherein oil phase and water separately (oil droplet can be coalescent).The key that agglomeration process is reduced to slight level is to add emulsifying agent in Emulsion.

For stoping or minimizing oxidation degradation reaction or lipid peroxidation, except deoxidation (for example places preparation noble gas such as nitrogen and argon, and/or use fast light container) in addition, perhaps as its alternative approach, can also add for example alpha-tocopherol and butylation benzylalcohol, and antiseptic such as edetate.

Can realize emulsifying by multiple technologies well-known in the art, as mechanical mixture, homogenize (as, adopt polytron or Gaulin high energy type equipment), vortex and supersound process.Supersound process can adopt water-bath type or sonde-type equipment to carry out.Microfluidization device for example can be available from Microfluidics Corp., Newton, and Mass. at United States Patent (USP) 4,533, further describes in 254, and utilizes the auxiliary narrow orifice of passing through of pressure.Also can the auxiliary extruding of working pressure pass the multiple polycarbonate membrane of buying.Low-voltage device also is operable.These high pressure and low-voltage device can be used to select and/or adjust the size of vesicle.

By the Filtration degerming.Filtration can comprise by than the pre-filtering of major diameter filter as 0.45 micron filter, and and then see through less filter, as 0.2 micron filter.Preferred filter medium is cellulose acetate (Sartorius-Sartobran ^TM).

Lyophilizing

Lyophilizing is from a sample liquid to be removed or removed basically, for example by literizations, is as described in the part of " removing of solvent " as title in the above.

The sign of physics and chemical stability and assessment

Phospholipid and catabolite thereof can extract from Emulsion and measure.Then liquid mixture is separated with two-dimentional thin layer chromatography (TLC) system or high performance liquid chromatography (HPLC) system.In TLC, can get the amount of measuring phosphorus corresponding to the speckle of single component.The total amount of phosphorus can quantitative assay in the sample, as measuring the intensity of the blueness of formation with respect to water at 825nm place with spectrophotometer.In HPLC, can separating phospholipids phatidylcholine (PC) and phosphatidyl glycerol (PG) and accurate quantification.Lipid can detect in the zone of 203-205nm.In the 200nm of ultraviolet spectra wavelength region may, unsaturated fatty acid shows higher absorbance, and satisfied fatty acid shows lower absorbance.As an example, Vemuri and Rhodes, the same, PC and PG in the egg yolk separating in Licrosorb Diol and LicrosorbS1-60 described.The mobile phase that separate to adopt is acetonitrile-methanol, and it contained hexane-water of 1% (74: 16: 10v/v/v).After 8 minutes, observation PG separates with PC's.Retention time is approximately 1.1 and 3.2 minutes respectively.

The outward appearance of Emulsion, average droplet diameter, and size distribution for observing and keeping, be important parameters.These parameters can adopt many methods to estimate.For example, can adopt dynamic light scattering and Electron Microscopy.Referring to, for example, Szoka andPapahadjopoulos, Annu.Rev.Biophys.Bioeng., 9:467-508 (1980).Especially, can adopt the freeze fracture electron micrograph to carry out morphology characterizes.Also can select the optical microscope of low usefulness for use.

The emulsion droplets size distribution for example can adopt the particle size distribution analyzer to detect, Horiba Limited (Ann Arbor for example, MI, USA) CAPA-500 of Zhi Zaoing, coulter counter (Beckman Coulter Inc., Brea, CA, or Zetasizer (Malvern Instruments, Southborough USA),, MA, USA).

The Detection of Stability of using HPLC to carry out

Similar to the above-mentioned method that is used for the Emulsion lipidic component, the chemical stability of therapeutic activity composition such as 17-AAG also can be assessed by HPLC after the extraction of Emulsion.Can develop special assay method, separate with its catabolite with Ansamycin with therapeutic activity.The degree of degraded can be assessed according to the minimizing of the HPLC peak-to-peak signal relevant with the therapeutic activity Ansamycin and/or according to the increase of the HPLC peak-to-peak signal relevant with catabolite.Compare with other components in the Emulsion component, Ansamycin detects maximum absorbance easily and clearly at the 345nm place.

Preparation and administering mode

Although administration all is preferred in various aspects of the present invention and embodiment medium-sized vein, but it will be appreciated by those skilled in the art that this method also can change or adapts to other administering modes at an easy rate, in for example oral, aerosol, non-gastrointestinal, subcutaneous, intramuscular, intraperitoneal, rectum, vagina, the tumor or tumor week administration.Next the major part of Tao Luning all is well known by persons skilled in the art, is used as background technology other probabilities of the present invention are described but still provide.Be appreciated that following content and top discussion will have eclipsed part.

Pharmaceutical composition can be by routine mixing, dissolving, granulation, make coated tablet, levigate, emulsifying, incapsulate, parcel or freeze drying process prepare.

The pharmacy acceptable composition can adopt conventional method to utilize one or more physiology acceptable carriers to prepare, and includes the excipient and the auxiliary agent that help reactive compound is processed into pharmaceutically acceptable preparation.Suitable compound method depends on the route of administration of selection.Some excipient and the existing description of the application in the preparation preparation thereof.Some other is known in the art, as PCT/US99/30631, Remmington ' s Pharmaceutical Sciences, MeadePublishing Co., Easton, PA (latest edition) and Goodman and Gilman ' s ThePharmaceutical Basis of Therapeutics, Pergamon Press, New York is described in the N.Y. (latest edition).

For injection, medicine can be prepared in aqueous solution, preferably in the physiology compatible buffers, prepares in hanks solution, Ringer's solution or the normal saline buffer solution as known in the art.For mucosal, in preparation, use the penetrating agent be suitable for the barrier that will penetrate.These penetrating agent are well-known in the art.

The above-mentioned preparation of the present invention and its all are very suitable for through injection as inject or continuous infusion carries out parenteral immediately or near administration immediately after the lyophilized cake hydration.Ejection preparation can be a unit dosage form, as is contained in ampoule or the multi-dose container, and adds antiseptic, as edetate.As previously discussed, pharmaceutical composition of the present invention can be stored under the inert environments, in the packing of or starvation fast light as ampoule or other.

Dosage range

The I phase pharmaceutical research of the 17-AAG that carries out in the solid tumor adult patient late, per three weeks have determined that by infusion every day administration in 1 hour 5 days maximum tolerated dose (MTD) is 40mg/m ²Wilson?et?al.，Am.Soc.Clin.Oncol.，abstract，Phase?IPharmacologic?Study?of?17-(Allylamino)-17-Demethoxygeldanamycin(AAG)in?Adult?Patients?with?Advanced?Solid?Tumors(2001)。In this research, the meansigma methods of t1/2, clearance rate and stable state capacity+/-the SD pH-value determination pH be 2.5+/-0.5 hour, 41.0+/-13.5L/ hour and 86.6+/-34.6L/m ²When dosage is 40 and 56mg/m ²The time, the C of blood plasma _MaxLevel be determined as respectively 1860+/-660nM and 3170+/-1310nM.As instructing, can expect that the useful patient dose scope of effective ingredient is about 0.40mg/m in the preparation of the present invention with these values ²To 4000mg/m ²M ²The expression body surface area.With canonical algorithm mg/m ²Change mg medicine/kg body weight into.

The following examples only are illustrative, unless particularly point out in the claims, component herein and step are not to limit the invention.Embodiment 1-5 and 9 has used for reference the application and has required priority, application on April 4th, 2003, title is the PCT that owns together the application PCT/US03/10533 of " novel Ansamycin preparation and preparation and using method ".

Embodiment

The preparation of embodiment 1:17-AAG; Alternative 1

Dropwise drip 36.0mL (470mmol) allylamine that is dissolved in the 50mL dry THF to the 45.0g that is dissolved in the 1.45L dry THF (80.4mmol) geldanamycin that places the dry flask of 2L, drip above 30 minutes.Reactant mixture in stirring at room 4 hours, the analysis showed that until TLC reaction finishes [GDM: glassy yellow: Rf=0.40 under nitrogen; (5%MeOH-95% CHCl ₃); 17-AAG: purple: Rf=0.42 (5% MeOH-95%CHCl ₃)].Remove by rotary evaporation and to desolvate, crude product under 25 ℃ at 420mL H ₂Form serosity among O: the EtOH (90: 10), filter then, and 45 ℃ dry 8 hours down, obtain 40.9g (66.4mmol) purple crystals 17-AAG (82.6% yield is monitored purity＞98% through HPLC at 254nm place).Recording fusing point with differential scanning colorimetry (DSC) is 206-212 ℃. ¹H NMR is consistent with the product that needs with the result that HPLC obtains.

Embodiment 2: the preparation of low melting point type 17-AAG

A kind of alternative purification process is the rough 17-AAG in 80 ℃ of dissolving embodiment 1 in 800mL 2-propanol (isopropyl alcohol), is cooled to room temperature then.Filter, descended dry 8 hours at 45 ℃ then, obtain 44.6g (72.36mmol) purple crystals 17-AAG (90% yield is monitored purity＞99% through HPLC at the 254nm place).Recording fusing point with differential scanning colorimetry (DSC) is 147-175 ℃. ¹H NMR is consistent with the product that needs with the result that HPLC obtains.

Embodiment 3: the preparation of low melting point type 17-AAG, alternative 1

A kind of alternative purification process is with the product 17-AAG of embodiment 2 H at 400mL ₂Form serosity in 25 ℃ among O: the EtOH (90: 10), filter, and 45 ℃ dry 8 hours down, obtain 42.4g (68.6mmol) purple crystals 17-AAG (95% yield is monitored purity＞99% through HPLC at 254nm place).Fusing point is 147-175 ℃. ¹H NMR is consistent with the product that needs with the result that HPLC obtains.

The preparation of embodiment 4:17-AAG Emulsion

The 17-AAG that the foregoing description 1-3 is obtained in any is dissolved in ethanol.Below form shows is a collection of goods according to the 4000gm of one embodiment of the invention preparation.One of ordinary skill in the art would recognize that this operating procedure can amplify or dwindle, the amount of each component also can change or the like, and also can add the component of not listing.

Component	??％w/w	Gram number in 4000g batch
Component	??％w/w	Gram number in 4000g batch	??17-AAG	??0.2	??8
??Miglyol?812	??15	??600	??17-AAG	??0.2	??8
??Miglyol?812	??15	??600	??Phospholipon?90G	??7.5	??300
The EDTA disodium, dihydrate, USP	??0.005	??0.2	??Phospholipon?90G	??7.5	??300
The EDTA disodium, dihydrate, USP	??0.005	??0.2	Sucrose, NF	??15	??600
Water for injection, USP	??QS	??QS	Sucrose, NF	??15	??600
Water for injection, USP	??QS	??QS	??0.2N?NaOH	Regulate pH to 6.0 ± 0.2	Add in a quantity as required

The polypropylene beaker that 17-AAG (CNF-101) is weighed and places 5L.Be incorporated as the about 50 times ethanol of drug weight, and place water-bath to carry out supersound process solution, with dispersion medicine.Then with Miglyol 812 (Sasol North America Inc; Houston, TX, USA) (CT USA) joins in the dispersion for American Lecithen Co., Oxford, mixture is placed to stir on the agitator disk up to solid roughly dissolved fully with Phospholipon 90G.Ultrasonoscope water-bath and/or be heated to about 45 ℃ and can help to carry out solid dissolving.Can check the level of dissolution of solution with optical microscope to guarantee to wish.

Under vigorous stirring, make dry air or nitrogen (National Formulary) gas communication cross liquid surface, reduce up to ethanol content, preferably be brought down below the 50%w/w of its initial amount with ethanol evaporation, more preferably less than 10%w/w, most preferably be about 5%w/w or lower.Solution checks being equipped with under the optical microscope of polarizing filter, with the level of dissolution of guaranteeing to wish, and preferably dissolving (not having crystallization or precipitate) fully.

(disodium, dihydrate USP), sucrose and water for injection (WFI) the polypropylene beaker of weighing and placing 5L, stir and dissolve until solid with EDTA.Then water is joined in the oil phase, fully mix up to being adhered to lip-deep oil with the speed of 5000RPM with the emulsify at a high speed device that has the Emulsion head and " peel off ".Then shearing is increased to 10000RPM 2-5 minute to obtain uniform primary emulsion.Laser light scattering (LLS) can be used to measure average droplet diameter, and solution can further detect, and determines crystal existence or the disappearance relative with solid under at optical microscope.

With the NaOH of 0.2N pH regulator to 6.0 ± 0.2 with Emulsion.Allow primary emulsion under the static pressure of about 110psi (operating pressure 60-95psi) by having 11OS type microfluidization device (the Microfluidics Inc. of 75 microns Emulsion application chambers (F20Y), Newton, MA, USA), by 6-8 time, until average droplet diameter≤190nm.At every turn by after can come its progress is assessed with LLS.Solution can further detect with polarized light whether the crystal existence is arranged under optical microscope.

Then in super quiet of laminar flow hood with Gelman microcapsule formula filter (the Pall Corp. of Emulsion by 0.45 micron, East Hills, NY, USA), then pass through 0.2 micron aseptic Sartorius Sartobran P bag type filter (500cm2) (Sartorius AG, Goettingen, Germany).Use up to the pressure of 60PSI and keep level and smooth and lasting flowing.Filtrate is collected in one or more polypropylene vial and is placed immediately-20 ℃ of refrigerators.Can reserve the equal portions of 1ml so that detect with laser light scattering (LLS) and/or high performance liquid chromatography (HPLC).

Embodiment 5: the preparation method of alternative 17-AAG emulsification preparation

When promoting that with ethanol 17-AAG is dissolved in the oil phase of Emulsion, the most frequently used is at first to utilize supersound process that 17-AAG is dissolved in the ethanol, then adds emulsifying agent and medium chain triglyceride again in this solution.Utilize supersound process then and stir the dissolving that realizes all components.

Selectable, can not use ethanol that 17-AAG is joined in the oil phase of solution, method comprises: heating preformed emulsifying agent in triglyceride solution, as

Phospholipon in 812,, preferably be heated to 65 ℃ or higher, to wherein adding medicine, as 17-AAG, and by mixing as stirring and/or supersound process.Have been found that at room temperature the low melting point type 17-AAG that is easier to prepare by crystallization 17-AAG in isopropyl alcohol rather than in ethanol among the embodiment 2 can be dissolved among the Phospholipon in the Miglyol solution.Referring to the title in JIUYUE in 2002 application on the 18th be the PCT/US01/29715 that owns together of " prepare ... .[17-AAG] and the method for other Ansamycins ".

Embodiment 5 and 6 product all are the lactous Emulsion of lilac, and the average diameter of oil droplet is about 200nm or littler.Preserve the time more than 2 months under 20 ℃, 2-8 ℃ or room temperature, the oil droplet size is stable.Totally reach concentration, and only in oil phase, its concentration is up to 20-30mg/ml up to 3mg/ml.If be stored under 40 ℃, can observe the degraded of 17-AAG after about two weeks.

The mixed solvent solution of medicine is through the Miglyol solution of vacuum evaporation ethanol component with formation 17-AAG.Emulsification can be handled, realize by microfluidization device at last by mechanical mixture, ultrasonic irradiation, yet be appreciated that, term " emulsifying " and " emulsification " are not limited in these processing methods, the emulsifying technology that also has other, and can select to use or use with one or more above-mentioned technical tie-ups.

Embodiment 6: add long chain triglyceride

The variation of said method comprises the adding long chain triglyceride, adds as the form with soybean oil.As among Fig. 1 about shown in the 17-AGG, the source of long chain triglyceride (soybean oil) and Miglyol 812N (source of medium chain triglyceride) and emulsifying agent (Phospholipon 90G (PL90G)) elder brother close, its w/w ratio is 16.7%: 50.0%: 33.3%.Carry out homogenize and dissolve (about 20, about 20 minutes of 000rpm) fully up to PL90G.The 17-AAG that adds 1%w/w then, it is about 20, and homogenize/dissolving is about 5 minutes under the 000rpm.Above-mentioned substance has constituted " oil phase ".Then this oil phase (1 part) is slowly joined 3.6 parts with about 12,000-15, the aqueous phase of 000rpm homogenize (being dissolved in the 9.375%w/w sucrose of sterile water for injection, 0.0063%w/w EDTA).In case of necessity the mixture of gained is regulated pH value to 6.0 ± 0.2 with sodium hydroxide and/or hydrochloric acid.Should carry out Micro Fluid by the F12Y application chamber by " just " Emulsion then, and with 0.2 μ m polyether sulfone (PES) membrane filtration degerming.

Adopt said method, prepare following two kinds of preparations:

Component (w/w) preparation 1 preparation 2

????????????????????????????????????????????????????????

17-AAG???????????????2mg/ml???????????2mg/ml

Soybean oil 3.3%---

Phospholipon?90G?????6.6％????????????7.5％

(soybean lecithin)

Miglyol?812N?????????9.9％????????????15％

Sucrose 7.5% 10%

Sodium ethylene diamine tetracetate 0.005% 0.005%

Sterilized water 72.5% 67.3%

Embodiment 7: lyophilizing

The lyophilizing of embodiment 5 and 6 Emulsion can be finished according to the scheme in the similar following table.

Under being stored in 2-8 ℃ and after rebuilding, the stability of lyophilizing 17-AAG Emulsion spectrum is as follows.

Embodiment 8: long chain triglyceride suppresses drowsiness

When quick administration, because metabolism discharges caprylate, Miglyol 812N can produce sedation.In the process of infusion 17-AAG Emulsion in to rat vein (Miglyol 812N oil), observe sedation during greater than the total fat of 1.1gm/Kg/hr at infusion velocity.Referring to accompanying drawing 2.In with Canis familiaris L., also observe sedation greater than the speed intravenous infusion 17-AAG emulsification preparation of the total fat/Kg/hr of 1.13gm.In order to resist this effect, add soybean oil as described above to compete, so that reduce the fatty acid caprylate that in the intravenous infusion process, produces with the internal metabolism of Miglyol 812N.For soybean oil/Miglyol 812NCF237 Emulsion, during up to the total fat of about 40gm/kg/hr, in rat, obviously do not observe sedation at infusion velocity.Therefore, the combination of soybean oil and Miglyol 812N has greatly improved the toleration of CF237 emulsification preparation to sedation.Similarly,, promptly do not observe sedation in the monkey of intravenous infusion 12mL preparation/kg/hr, do not observe venous stimulation yet using the CF237 emulsification preparation of six multiple doses.

Embodiment 9: be used for the preparation of other Ansamycin of similar formulations

Ansamycin beyond the 17-AAG

Basically all Ansamycins can replace 17-AAG, and according to the described preparation of top embodiment.Multiple such Ansamycin and their preparation are described in detail in PCT/US03/04283.Wherein two kinds preparation is described below.

Chemical compound 563:17-(benzoyl)-amino geldanamycin mycin.EtOAc solution Na with 17-amino geldanamycin mycin (1mmol) ₂S ₂O ₄(0.1M 300ml) at room temperature handles.After 2 hours, water layer extracts twice with EtOAc, and blended organic layer is at Na ₂SO ₄Last dry, under low pressure concentrate the back and obtain yellow solid 18,21-dihydro-17-amino geldanamycin mycin.The latter is dissolved in anhydrous THF, again by intubate transfer to Benzenecarbonyl chloride. (1.1mmol) and

In the mixture (1.2g).After 2 hours, with EtN (i-Pr) ₂(2.5mmol) join in the reaction mixture.After stirring was spent the night, filter reaction mixture also under low pressure concentrated.Add water then in residue, extract three times with EtOAc, blended organic layer is at Na ₂SO ₄The last dry and under low pressure concentrated crude product that obtains, crude product has obtained 17-(benzoyl)-amino geldanamycin mycin through purified by flash chromatography.CH at 80: 15: 5 ₂Cl ₂: EtOAc: among the MeOH, Rf=0.50.Mp＝218-220℃。1H?NMR(CDCl ₃)0.94(t，6H)，1.70(br?s，2H)，1.79(br?s，4H)，2.03(s，3H)，2.56(dd，1H)，2.64(dd，1H)，2.76-2.79(m，1H)，3.33(br?s，7H)，3.44-3.46(m，1H)，4.325(d，1H)，5.16(s，1H)，5.77(d，1H)，5.91(t，1H)，6.57(t，1H)，6.94(d，1H)，7.48(s，1H)，7.52(t，2H)，7.62(t，1H)，7.91(d，2H)，8.47(s，1H)，8.77(s，1H)。

Chemical compound 237: a kind of dimer.At N ₂Following 3 of condition, (1.32g, (10g in DESO 17.83mmol) (200ml) solution, stirs under the room temperature 3-diaminourea-di-n-propylamine 9.1mmol) dropwise to join the geldanamycin that places flame-dried flask.Dilute with water reactant mixture after 12 hours.Reactant forms precipitation, and filters to obtain crude product.Crude product is through silicon chromatography (5%CH ₃OH/CH ₂Cl ₂) chromatography, obtain to be solid this dimer of purple.Behind flash chromatography (silica gel) purification, obtain the pure products of purple; Yield: 93%; 165 ℃ of mp; 1H NMR (CDCl ₃) 0.97 (d, J=6.6Hz, 6H, 2CH3), 1.0 (d, J=6.6Hz, 6H, 2CH3), 1.72 (m, 4H, 2CH2), 1.78 (m, 4H, 2CH2), 1.80 (s, 6H, 2CH3), 1.85 (m, 2H, 2CH), 2.0 (s, 6H, 2CH3), 2.4 (dd, J=11Hz, 2H, 2CH), 2.67 (d, J=15Hz, 2H, 2CH), 2.63 (t, J=10Hz, 2H, 2CH), 2.78 (t, J=6.5Hz, 4H, 2CH2), 3.26 (s, 6H, 2OCH3), 3.38 (s, 6H, 2OCH3), 3.40 (m, 2H, 2CH), 3.60 (m, 4H, 2CH2), 3.75 (m, 2H, 2CH), 4.60 (d, J=10Hz, 2H, 2CH), 4.65 (Bs, 2H, 2OH), 4.80 (Bs, 4H, 2NH2), 5.19 (s, 2H, 2CH), 5.83 (t, J=15Hz, 2H, 2CH=), 5.89 (d, J=10Hz, 2H, 2CH=), 6.58 (t, J=15Hz, 2H, 2CH=), 6.94 (d, J=10Hz, 2H, 2CH=), 7.17 (m, 2H, 2NH), 7.24 (s, 2H, 2CH=), 9.20 (s, 2H, 2NH); MS (m/z) 1189 (M+H).

Corresponding hydrochlorate is prepared by following method: (5ml, 0.123N) solution joins among the THF (15ml) and EtOH (50ml) solution of chemical compound #237 (1gm of above-mentioned preparation) with the EtOH of HCl under the room temperature.Reactant mixture was stirred 10 minutes.Salt is precipitated out, and refilters and with a large amount of EtOH washing, vacuum drying.

*????????????????*????????????????*

Whether those of ordinary skill is appreciated that the parameter in the foregoing description and the form can adjust according to the condition that adopts, and amplify or dwindle or how change respect to one another and degree are adjusted according to the amount of compound method and material therefor.

The foregoing description is not to be restrictive, and they just represent different aspect of the present invention and embodiment.Represented the technical merit in field under the present invention at all documents that this quotes.Although the document of quoting does not have one piece to be considered to prior art, the content of every piece of quoting document is all quoted as a reference with same degree at this, is all quoted as a reference fully independently as every piece of document.If clearly defining and appearing at prior art or the definition in the priority text that this quotes and have and conflict herein, then the clearly definition with the application is as the criterion.

Those skilled in the art will realize the inherent characteristics that the present invention can reach above-described target well and obtain The above results, advantage and comprise at an easy rate.Described method and composition is used for illustrating preferred embodiment, is exemplary rather than limitation of the scope of the invention.Those skilled in the art will envision that some change and other purposes, these are changed and other purposes is included in by in the defined scope of the present invention of claims scope.

Described reagent both can be buied, and as available from Sigma-Aldrich, also can need not conventional method too much test just can easily prepare by well known to a person skilled in the art.

It will be apparent to one skilled in the art that under the situation that does not depart from spirit and scope of the invention, can carry out multiple replacement and change the present invention.Therefore, other embodiment of this class is all in the scope of the present invention and following claims.

Lacking under the situation of this not concrete disclosed any key element, restriction, also can suitably be implemented in the present invention that this illustrative is described.For example, under all situations herein, term " comprises ", " substantially by ... form " and " by ... form " each can be with the replacement of one of two other term, and each term all has different implications on Patent Law.Term that uses and statement are to limit as descriptive term rather than with it, the use of these terms and statement do not get rid of of the present invention and shown in any equivalent feature or its partial content.One skilled in the art will recognize that simultaneously it is fully possible carrying out multiple change in the scope of protection of present invention.Therefore be appreciated that, though the present invention is by preferred embodiment and optionally feature is specifically disclosed, but those skilled in the art can carry out modifications and changes to notion disclosed herein, and these modifications and changes are considered in description and the defined scope of the present invention of claim.

In addition, feature of the present invention or aspect are described with Ma Kushi (Markush) group or other alternative group, those skilled in the art can think that the also available Ma Kushi group of the present invention or each member of other group or member's subgroup are described, and get rid of by each member's qualifications in case of necessity.

Other embodiment within the scope of the following claims.

Claims

1. pharmaceutical composition, it comprises:

The HSP90 inhibitor, optional Ansamycin;

Emulsifying agent; With

Oil, described oil comprises medium chain triglyceride and long chain triglyceride simultaneously.

2. the pharmaceutical composition of claim 1, wherein said HSP90 inhibitor is an Ansamycin.

3. the pharmaceutical composition of claim 2, wherein said Ansamycin is geldanamycin or geldanamycin derivant.

4. the pharmaceutical composition of claim 3, wherein said geldanamycin derivant is selected from 17-AAG and DMAG.

5. the pharmaceutical composition of claim 1, wherein said medium chain triglyceride is 10: 1 to 0.01: 10 with the w/w ratio of described long chain triglyceride.

6. the pharmaceutical composition of claim 1, it is to have fat mutually and the oil in water emulsion of water, wherein said fat accounts for the 5-30%w/w of gross weight mutually.

7. the pharmaceutical composition of claim 1, it comprises the phospholipid of 3-10%w/w and the oil of 5-20%w/w.

8. the pharmaceutical composition of claim 7, wherein said phospholipid exists with the lecithin form.

9. the pharmaceutical composition of claim 8, wherein the amount of lecithin is the 3-10%w/w of oil in water emulsion.

10. the pharmaceutical composition of claim 1, it comprises the long chain triglyceride that is no more than 10%w/w.

11. the pharmaceutical composition of claim 10, it further comprises the long chain triglyceride that is less than or equals 7%w/w.

12. the pharmaceutical composition of claim 10, wherein said long chain triglyceride comprises the fatty acid ester of being made up of 16-18 straight chain carbon unit, and described triglyceride is selected from one or more in linoleic acid, oleic acid, Palmic acid and the stearic acid.

13. the pharmaceutical composition of claim 7, wherein said phospholipid are the lecithin forms, optional is Semen sojae atricolor or egg lecithin.

14. being fusing points, the pharmaceutical composition of claim 2, wherein said Ansamycin be no more than 200 ℃ 17-AAG low melting point isomer.

15. the pharmaceutical composition of claim 14, wherein said low melting point isomer are fusing points is 147-175 ℃ 17-AAG.

16. the pharmaceutical composition of claim 1, wherein said oil comprise one or more natural oil that is selected from soybean oil, Oleum sesami, safflower oil and Semen Maydis oil.

17. the pharmaceutical composition of claim 1, it is a kind of Emulsion, and optional is freeze dried.

18. the pharmaceutical composition of claim 1, it comprises in water, antiseptic, antifreeze, buffer, chelating agen and the tonicity contributor one or more.

19. the pharmaceutical composition of claim 1, it comprises Miglyol 812N.

20. each described pharmaceutical composition of claim 1-19 wherein comprises the 17-AAG of 0.5mg/ml-4mg/ml.

21. each described pharmaceutical composition of claim 1-19 wherein comprises the 17-AAG of 0.05%w/w-0.4%w/w.

22. the pharmaceutical composition of claim 1 wherein comprises the 17-AAG of following component: 2mg/ml, 3.3% soybean oil, 6.6% lecithin, 9.9% Miglyol 812N, 7.5% sucrose, and water.

23. the pharmaceutical composition of claim 1 wherein comprises the 17-AAG of following component: 2mg/ml, 7.5% lecithin, 15% Miglyol 812N, 10% sucrose, and water.

24. the pharmaceutical composition of claim 22 or 23, it further comprises sodium ethylene diamine tetracetate.

25. the pharmaceutical composition of claim 24, wherein said sodium ethylene diamine tetracetate are 0.005%w/w.

26. the pharmaceutical composition of claim 1, the amount of wherein said long chain triglyceride enough reduce or avoid taking place central nervous system's effect of medium chain triglyceride mediation.

27. the pharmaceutical composition of claim 26, wherein said central nervous system's effect is selected from one or more in drowsiness, nauseating, sleepy and the EEG change.

28. the pharmaceutical composition of claim 1, it further comprises short chain triglyceride.

Accept to comprise medium chain triglyceride and as the patient of the pharmaceutical preparation of formulation components purposes by the medicine of central nervous system's effect of medium chain triglyceride mediation takes place 29. a pharmaceutical preparation is used for reducing in preparation, the amount that wherein said pharmaceutical preparation comprises Ansamycin and medium chain and long chain triglyceride and described long chain triglyceride is enough to reduce or avoid the generation of central nervous system's effect of medium-chain fatty acid mediation.

30. the purposes of claim 29, wherein said central nervous system's effect is selected from one or more in drowsiness, nauseating, sleepy and the EEG change.

31. the pharmaceutical composition of claim 1, it is in lyophilizing, freezing, thawing or the reconstruction form one or more.

32. the pharmaceutical composition of claim 1, it is stored in the inert environments.