CN102526753A - In-situ phase change gel slow release system taking phospholipid as substrate and preparation method thereof - Google Patents

Abstract

The invention provides an in-situ phase change gel slow release system taking a phospholipid as a substrate, and provides a preparation method thereof. A high-concentration phospholipid slow release preparation is prepared from a high-concentration (50-85 percent) phospholipid, a bioactive ingredient, ethanol solutions of different concentrations and/or injection oil with a simple method, and has the characteristics of high biocompatibility, small untoward effect, remarkable slow release effect and suitability for various administration forms such as hypodermic injection, intramuscular injection, external administration and the like; the amount of a coated medicament can be conveniently adjusted according to the clinical dosage of a medicament; and the in-situ phase change gel slow release system has a wide application prospect.

Description

A kind of is the original position phase change gel slow-released system and preparation method thereof of substrate with phospholipid

Technical field

The present invention relates to a kind of in-situ injection phase change gel slow-released system, be specifically related to be prepared into high concentration phospholipid slow releasing preparation, be medical technical field with phospholipid, Different concentrations of alcohol solution and/or oil for injection.

 

Background technology

Along with the rise and the development of biotechnology, the diagnosis and the treatment of disease developed and be used for to increasing protein and peptide drugs.Compare with the micromolecule chemicals, normal physiologically substance is more approaching in protein and peptide drugs and the body, has the pharmacologically active height, and toxic and side effects is low, and dosage is little, determined curative effect, advantages such as high specificity.But also exist a lot of problems simultaneously: on the one hand, the protein and peptide drugs molecular mass is big, and poor stability easily by the Proteolytic enzyme enzymatic degradation in the gastrointestinal tract, therefore, can not adopt common gastrointestinal administration mode usually, can only pass through drug administration by injection; On the other hand, the biological half-life of this type medicine is shorter, is decomposed by body soon, and the curative effect that needs lasting injection or intravenous drip to guarantee medicine has increased the weight of patient's body, psychology and financial burden.

Therefore, people hope to develop a kind of Atrigel of injectable protein and peptide drugs.

At present, the Injectable sustained release drug-supplying system that has gone on the market mainly comprises PLGA (polylactic acid-glycolic guanidine-acetic acid copolymer) microsphere and multivesicular liposome (MVL).Microsphere is meant medicine dissolution or is dispersed in the small spherical entity that forms in the macromolecular material substrate, belongs to matrix type skeleton microgranule.PLGA is the copolymer that is polymerized by a certain percentage by lactic acid and hydroxyacetic acid, is the Biodegradable material that can be used for human body of FDA approval.

In many polypeptide slow releasing injection, LHRH (luteinizing hormone releasing hormone) analog microsphere is to study the most successful kind.Triptorelin is one of LHRH analog, and its PLGA microsphere is by the exploitation of French Ipsen company, and listing in 1986, but slow release 1 month are first polypeptide microspheres products.Alarelin is the first class national new drug of Shanghai beautiful pearl east wind Bioisystech Co., Ltd exploitation; Curative effect is 15 times of LHRH; Shanghai Institute of Pharmaceutical Industry bears State Scientific and Technological Commission " 95 " brainstorm project; Carry out the developmental research of the continuous release microsphere of medication in every month 1 time, its slow release effect of animal vivo test proof is reliable.

Although the PLGA microsphere has slow-releasing and controlled-releasing action and biocompatibility preferably; But because its complicated process of preparation; Drug loading is lower; The organic solvent that uses in the preparation process has residual in preparation, and produces lactic acid in the degradation process and hydroxyacetic acid can cause the decline of injection site pH value, thereby produces local aseptic inflammation.The big limitations of above shortcoming and defect the application of PLGA microsphere.

Multivesicular liposome (MVL) is the special novel lipid body preparation of a kind of structure, and its discontinuous non-concentric aqueous inner chamber is cut apart through lipoid and immobilized artificial membrane, forms a lot of vesicles.In this discontinuous vesicle, when certain vesicle broke, medicine only disengaged from disruptive vesicle, and complete vesicle still maintains the original state, thereby had the good slow release effect.The height that traditional liposomal generally is difficult to reach water soluble drug with prior art is sealed and is hanged down seepage; Multivesicular liposome has advantages such as envelop rate height, drug leakage are few by comparison, is particularly useful for sealing soluble small molecular medicine and bioactive macromolecule medicine.

At present; SkyePharma company utilizes DepoCytTm (cytosine arabinoside MVL preparation) and two products of DepoDurTm (morphine sulfate MVL preparation) of DepoFormTm technology (registered trade mark of MVL technology) exploitation successfully to go on the market, and has a plurality of products to get into research and development and clinical research stage.

Although the multivesicular liposome route of administration is various, have good slow release effect and bank effect, it also has many weak points: at first, desire prepares MVL, need contain parents' lipid (like phospholipid) and neutral lipid (like triglyceride) at least.If no neutral lipid then forms traditional unilamelar liposome (ULV) or multilamelar liposome (MLV).Secondly; Because the inside and outside water of MVL exists drug level poor, the leakage problems of small-molecule drug still exists, and phospholipid bilayer also partial rupture can occur when storing; And it is not resolved in sedimentation and rendezvous problem that storage process occurs as yet, possibly influence stability of formulation.The 3rd, the MVL product of exploitation is a suspension at present, is not easy to storage and transport.The 4th, the preparation process of MVL is complicated, and working condition is required very harshness, is difficult for enlarging and produces.In addition, the medication amount that the MVL bag carries is limited, and this also makes its development receive certain restriction.

In addition, and the vesicle type phospholipid gel of studying in addition (vesicular phospholipid gels, VPG).VPG is a kind of semisolid phospholipid dispersion, has unique three-dimensional netted stereochemical structure, is different from traditional lipidosome gel and conventional liposome cell structure.VPG is as the bank of medicine, in the substrate of drug distribution outside vesicle and vesicle.Under the situation of water as solvent, all contain the water of certain volume in the vesicle and between vesicle, but loaded with water-soluble, fat-soluble and amphipathic medicine.For water soluble drug; The inside and outside water volume that equates that is close to of vesicle not only makes its envelop rate improve; And medicine all exists in vesicle with outside the vesicle, and interior extracellular concentration reaches unanimity, weakened to a certain extent since the drug leakage that Concentraton gradient causes etc. to the influence of stability.

Now existing more document has disclosed about the report of vesicle type phospholipid gel as the peptide medicament Atrigel.For example; Brandl; The research group at people such as Winter place adopt micromolecule chemicals calcein and 5-fluorouracil, micromolecule polypeptide cetrorelix respectively and with the high molecular weight protein erythropoietin as model drug; In homemade release flow cell research with VPG release in vitro behavior (C. Tardi, Journal of Controlled Release 55 (1998) 261 – 270 during as drug depot; N. Kaiser, International Journal of Pharmaceutics 256 (2003) 123 – 131; H. Grohganz, European Journal of Pharmaceutics and Biopharmaceutics 59 (2005) 439 – 448; W. Tian, Journal of Controlled Release 142 (2010) 319 – 325).Its experimental result shows that VPG is better to the slow release effect of high molecular weight protein, can reach hundreds of hours; Discover in the body of patent application 20110036587.8 through animal; Concentration also has long slow-release time at the phospholipid gel of 20-40% for micromolecule polypeptide class medicine; But the phospholipid gel subcutaneous injection initial stage burst effect of low concentration is obvious, might cause toxic and side effects.Along with the increase of phospholipid concentration, prominent releasing reduces.When phospholipid concentration was elevated to 40%, though slow release effect is better, its viscosity reached 14.3PaS (W. Tian, Journal of Controlled Release 142 (2010) 319 – 325), and quite thickness almost can not be injected.So can find out from existing research, vesicle type phospholipid gel not too is suitable for as injection polypeptide slow-released carrier.

Patent documentation 93119112.2 has disclosed a kind of lecithin gel that forms in vivo to continue the Pharmaceutical composition of delivery of biologically active chemical compound, comprises lecithin, pharmaceutically acceptable confession intramuscular or subcutaneous injection and water-fast basically lecithin with solvent and bioactive compound.Briefly, this Pharmaceutical composition is exactly lecithin organic solution or the suspension that contains bioactive compound.Through intramuscular or this solution of subcutaneous injection or suspension, can form the lecithin gel in vivo, promptly form the lecithin gel in vivo through absorbing moisture the body fluid between moisture crack, injection site.According to patent description, medicine length of release time from compositions is decided by the composition of said composition.In addition, can also control through adding excipient.Yet owing to lack polar solvent in the main component of said composition, there is certain degree of difficulty in the medicinal application that water soluble drug or polarity are stronger in said composition.Although say in the document and can active component is dissolved in earlier in the low amounts of water or be fit in the buffer solution of this concrete active component, and then water or buffer are added in the compositions, just before injection, increased the viscosity of compositions like this, be unfavorable for drug administration by injection.

Patent documentation 200580025364.4 provides a kind of compositions that comprises phospholipid fraction and the acceptable fluid carrier of medicine; But this application is smudgy; As during the localized drug delivery that is used to continue; It is 10-90% that said composition limits phospholipid fraction concentration in certain embodiments, and maximum quantity is 45% at the most in certain embodiments, is which embodiment but do not offer some clarification on.Prepared compositions among the embodiment 1 is because the dissolubility of phospholipid in selected solvent is not high; In order under suitable viscosity, to be injected into human body smoothly; Phospholipid concentration is all lower; And the large usage quantity of propylene glycol, glycerol equal solvent, even so said composition inject human body on a small quantity, also can cause local swelling even fester.In addition; Its process is: weigh up and mixing soya beans lecithin (Phospholipon 90G in the container of cleaning; The injectable grade soybean lecithin that comprises about 90% phosphatidylcholine, Phospholipid GmbH), medium chain triglyceride (Miglyol812, Sasol Corp.), sucrose and propylene glycol; Add dehydrated alcohol, the dissolving all solids is to form limpid yellow solution.Evacuation is to remove ethanol, up to remaining ethanol content less than 5% of gross weight.Heat this mixture to 60 ℃ to form clear solution, filter this solution through sterilization filter then.The cool to room temperature of will filtrating obtains yellow transparent and even gel, and what this shows the above-mentioned phospholipid of dissolving is not ethanol but propylene glycol.

More crucially, this application is not carried out experiment in the body, and it can reach the slow release effect of expection not have digital proof; Those skilled in the art are from the formation of its lower phospholipid concentration; Less volume injected in conjunction with the known trial of prior art, can assert that fully its slow releasing function is very poor.

Therefore, develop and a kind ofly can be expelled to the slow release effect that reaches good in the body, a difficult problem and focus that viscosity is lower again, phospholipid slow releasing preparation drug administration by injection smoothly just becomes this area research.

Summary of the invention

One of the object of the invention provides a kind of in-situ injection phase change gel slow-released system.

Be surprised to find that under study for action the dissolubility of phospholipid in ethanol is high especially, even work as the content of phospholipid in ethanol up to 85% (g/g), still clear can flow.According to this character, inventor imagination, with medicine dissolution in the alcoholic solution of this high concentration phospholipid; After injecting in the body; Because the second alcohol and water can dissolve each other fully, after ethanol was diffused into body fluid rapidly, then the remaining phospholipid in injection site can solidify; Thereby become the carrier of medicament slow release, effectively control the release of medicine.

Gel formation process of the present invention also be with prior art in the common phospholipid gel mentioned different fully.Patent documentation 93119112.2 has disclosed lecithin organic solution or the suspension that contains bioactive compound; This organic solution is water insoluble basically; After intramuscular or subcutaneous injection, form the lecithin gel in vivo through absorbing moisture the body fluid between moisture crack, injection site again.

According to above-mentioned unexpected discovery, the inventor is a model drug with the octreotide acetate thus, has prepared the alcoholic solution that contains 70% phospholipid, and to behind the rat skin lower injection 1ml, medicine can steadily discharge almost about one month.But adopt single ethanol as solvent, side effect also is extremely tangible, and red and swollen and the phenomenon of festering appear in most of rat injection site.This also is to avoid the use of a large amount of ethanol in the prior art as one of major reason of injection solvent as far as possible.

In order to overcome alcoholic acid side effect; Through creationary research, we are surprised to find, if in preparation, replaced part ethanol with soybean oil; The redness and the phenomenon of festering no longer appear in the rat injection site, and this is the effective solution that helps the industrialization of this slow releasing preparation system.

One of the object of the invention provides a kind of compositions that comprises phospholipid, alcoholic solution and/or oil for injection.

Through more deep research, the inventor finds, in solvent, adds oil for injection, is to reduce consumption of ethanol on the one hand; On the other hand, reduce alcoholic acid diffusion velocity, thereby not only realized effective utilization of phospholipid gel slow-released system, also reduced alcoholic acid side effect widely.

One of the object of the invention provides and a kind ofly is prepared into high concentration phospholipid slow releasing preparation with phospholipid, alcoholic solution and/or oil for injection, in-situ injection phase change gel slow-released system promptly of the present invention or in-situ injection phase change gel slow releasing preparation.

Can know according to the specific embodiment of the present invention; A series of liquid fat acid glycerol three esters comprise medium chain length fatty acid triglyceride, Oleum Sesami, Semen Maydis oil, Oleum Gossypii semen, fish oil etc., can both replace part ethanol as soybean oil; Reduce alcoholic acid side effect, finally realized the present invention thus.

According to the specific embodiment of the present invention, oil for injection of the present invention includes, but not limited to the combination of soybean oil, Semen Maydis oil, Oleum Sesami, Oleum Camelliae, fish oil, medium chain length fatty acid triglyceride, Oleum Gossypii semen, ethyl oleate and above-mentioned substance.

Though have the technical scheme that comprises lecithin, oil for injection, ethanol, active component in the prior art, the concrete preparation of these compositionss generally is an Emulsion, the mechanism of action is different from original position phase change gel preparation of the present invention fully.

One of the object of the invention provides a kind of compositions that comprises active constituents of medicine, phospholipid, alcoholic solution and/or oil for injection.After said composition was injected in the body, ethanol spread rapidly, and the phospholipid of separating out forms gel, and the packaging medicine active component has the good slow release effect.

One of the object of the invention provides a kind of had good sustained release effect, high concentration phospholipid content, is easy to the phospholipid slow releasing preparation that contains bioactive ingredients of drug administration by injection.

One of the object of the invention is that protein and peptide drugs or other drug are dissolved in the phospholipid-alcoholic solution of high concentration, becomes the liquid of good fluidity; After injecting in the body, ethanol spreads rapidly, and the phospholipid of separating out forms gel; Packaging medicine has the good slow release effect.

Term among this paper " in-situ injection phase change gel slow-released system " also comprises high concentration phospholipid slow releasing preparation.

Be applicable to the phospholipid of high concentration phospholipid slow releasing preparation of the present invention, include but not limited to: one or more combination in natural phospholipid, semi-synthetic phospholipid, the synthetic phospholipid.

Said natural phospholipid includes but not limited to Ovum Gallus domesticus Flavus lecithin, soybean phospholipid or its combination etc.

Said semi-synthetic phospholipid includes but not limited to hydrogenated yolk lecithin, hydrogenated soya phosphatide or its combination etc.

Said synthetic phospholipid includes but not limited to one or more the combination etc. in two palmityl PHOSPHATIDYL ETHANOLAMINEs, dipalmitoyl phosphatidyl choline, DSPC, dimyristoyl phosphatidyl choline, DOPE, two palmityl phosphatidyl glycerols, the two palmityl phosphatidic acid.

In concrete embodiment, the preferred Ovum Gallus domesticus Flavus lecithin E80 of the present invention.

In concrete embodiment, high concentration phospholipid slow releasing preparation of the present invention, based on the gross weight meter of preparation, wherein content of phospholipid is about 50%～about 85% (g/g).

Be applicable to the bioactive ingredients of preparation high concentration phospholipid slow releasing preparation of the present invention; Can be water-soluble molecules, lipophilic molecule or amphipathic molecule, include but not limited to: anticarcinogen class, anti-inflammatory agent class, anti-infective class, analgesic class, hormones, antidiabetic drug class, antihypertensive class, anti-acquired immunodeficiency syndrome drug class, immunostimulant class, antiviral agents class, cardiac tonic class, antiadipositas drug class, bone metabolism regulator class, antuepileptic class, anticonvulsant class, antidepressants class, psychosis class, antiparkinsonism drug class, urinary tract medicine class, contraceptive class, anti-osteoporotic class, anabolic agents class, smoking cessation auxiliary agent class and cell attachment promoter class.

Be applicable to that medicine of the present invention includes but not limited to as follows:

Protein and peptide; Include but not limited to that insulin, glucagon, alpha-interferon, beta-interferon, interleukin, erythropoietin (EPO), GTH, granulocyte-macrophage colony stimutaing factor (GM-GSF), tissue-type plasminogen activator, streptokinase, cell growth factor, thrombin, prostaglandin, antithrombase, hirudin, histrelin, GHRP-2, GHRP-6, cetrorelix, bivalirudin, arginine vasopressin, Schweine-Vasopressin, Desmopressin, gonadorelin, leuprorelin, De She Rayleigh, buserelin, triptorelin, goserelin, my Rayleigh, Sermorelin, nafarelin, lutrelin, fertirelin, sea sand Rayleigh, octreotide, Exenatide, human glucagon-like-peptide-1, growth hormone, somatostatin, ziconotide, thymosin a1, sincalide, Thymopentin, 1: PN: WO02056903 PAGE: 25 claimed protein, aviptadil, LEK, methionine enkephalin,MEK, tetracosactide, gonadotropin releasing hormone, throtropin releasing hormone, growth hormone releasing hormone (GHRH), Somat (GHIH), short melanocyte inhibitory hormone (MRIH), short melanocyte releasing hormone (MRH), corticotropin-releasing hormone (CRH), secretin, P material, neurotensin, human brain natriuretic peptide, ANP, hypertensin, angiotensin II, angiotensin II I, sleep-inducing peptide, hypermnesia peptide, dynorphin, endorphins, angiotensin, En Fuwei ground, CJC-1295, elcatonin, thymosin β 4, vassopressin, Pitressin Tannate, felypressin, ornipressin, melanotan II, spleen pentapeptide, AC-137, vapreotide, thyroliberin, silk rely short cortex 18 peptides, short cortex 18 peptides of sweet essence, short cortex 24 peptides of zinc, short cortex 24 peptides, short cortex 25 peptides, urge cortex 28 peptides, vasoactive intestinal peptide, BNF, Kallidin I, OFQ, asparaginase etc.; Micromolecule chemicals class includes but not limited to, semi-annular jade pendant reaches liver sodium in the last of the ten Heavenly stems, morphine, tramadol, oxycodone, fentanyl, lignocaine, dolantin, sorbide nitrate; Pentaerithrityl tetranitrate, isosorbide mononitrate, general naphthalene Nore is received many Nores, indole Nore, Ah 's Nore, Mei Tuonuoer, A Pu Nore, vinegar fourth Nore, nifedipine; Verapamil, diltiazem, spectrum Ni Laming, but piperazine former times woods, bepridil, nicorandil, molsidomine, dilazep, trimetazidine, quinidine sulfate; Third pyridine, the medicine of mexiletine hydrochloride, Amiodarone Hydrochloride, lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, cerivastatin, Xuezhikang; Colestyramine, colestipol, probucol, gemfibrozil, aspirin, ozagrel, ticlopidine, clopidogrel, ridogrel, tirofiban; Warfarin, dicoumarol, acenocoumarol, aesculetin, angelica lactone, astragaloside, ginsenoside, auraptene, furocoumarin, pyranocoumarin; Pteryxin, calophylloide, bisaesculetin, silibinin, puerarin, rutin, Hesperidin, catechin, Herba Pelargonii Graveolentis glycosides, breviscapine; Quercetin, resveratrol, Li Keding, captopril, enalapril, lisinopril, benazepril, fosinopril, ramipril, quinapril; Perindopril, cilazapril, trandolapril, alacepril, delapril, losartan, irbesartan, Candesartan, nifedipine, amlodipine; Felodipine, nitrendipine, lacidipine, nimodipine, nicardipine, nisoldipine, isradipine, diltiazem, verapamil, Propranolol; Mei Tuonuoer receives many Nores, indole Nore, Ah 's Nore, Mei Tuonuoer, A Pu Nore, vinegar fourth Nore, peso Nore, his Nore doubly, prazosin; Terazosin, doxazosin, trimazosin draws the shellfish Nore, screens Nore, clonidine, methyldopa, moxonidine, reserpine, guanethidine; Hydralazine, sodium nitroprusside, donepezil hydrochloride, Rivastigmine, huperzine A, metrifonate, tacrine, galantamine, piracetam, almitrine, methanesulfonic acid dihydroergocristine, pentoxifylline, nicergoline; Vitamin E melatonin, curcumin, desferrioxamine, idebenone, Tirilazad Mesylate, estradiol, ethinylestradiol, estrone, quinestrol, nilestriol, estradiol valerate, estradiol benzoate, androstenediol, methyltestosterone, testosterone, diethylstilbestrol; The diethylstilbestrol propionic ester, diethylstilbestrol phosphate ester, estradiol cypionate, big legumin, medroxyprogesterone acetate, megestrol acetate, CA, gestodene, norethindrone; Desogestrel, acid in the sixth of the twelve Earthly Branches progesterone, levonorgestrel, norethindrone acetate, norethisterone enanthate, dibasic acid esters etynodiol, Quingestanol Acetate, ground norgesterone, drospirenone; The alkene norgesterone, nomegestrol, trimegestone, diphenhydramine, promethazine, tripelennamine, chlorphenamine, buclizine, phenindamine; Cyproheptadine, clemastine, azelastine, astemizole, loratadine, cetirizine, mizolastine, teldane The Fexofenadine fourth, Efletirizine, Desloratadine, ebastine, Promethazine, pheniramine, brompheniramine, tolpropamine, pyrrobutamine, the sharp pyridine of the music score of Chinese operas, acrivastine, cimetidine, ranitidine, famotidine; Nizatidine, Luo Sha is for fourth, ketotifen, sodium cromoglicate, cyclophosphamide, ifosfamide, carmustine, lomustine, semustine, dacarbazine; Procarbazine, temozolomide, chlorambucil, melphalan, platinum complexes (like cisplatin, carboplatin, oxaliplatin etc.), methotrexate, fluorouracil; Ftorafur, mercaptopurine, pentostatin, cytosine arabinoside, gemcitabine, vinblastine, vincristine, vinorelbine, vindesine; Paclitaxel, docetaxel, homoharringtonine, harringtonine, camptothecine, hydroxycamptothecin, Top is for health, irinotecan, etoposide; Teniposide, actinomycin D, mitomycin, bleomycin, daunorubicin, doxorubicin, idarubicin, aclarubicin, mitoxantrone; Epirubicin, plicamycin, adrenocortical hormone, androgen, flutamide, estrogen, tamoxifen, exemestane, tretinoin; Imatinib, gefitinib, erlotinib, Thalidomide, Rayleigh degree amine, aspirin, magnesium salicylate, choline salicylate, salicylamide; Salsalate, diflunisal, indomethacin (indometacin), sulindac, acetaminophen (acetaminophen), aminophenazone, oxyphenbutazone, Phenylbutazone, mefenamic acid; Clofenamic acid, diclofenac, ibuprofen, naproxen, the fluorine ratio is coughed up sweet smell, fenoprofen, ketoprofen, piroxicam, meloxicam; Nimesulide, rofecoxib, celecoxib, colchicine, probenecid, other purine, sulfinpyrazone, benzbromarone etc., or the pharmaceutically acceptable salt form of the above-mentioned medicine of mentioning.

Further, above-mentioned protein and peptide drugs or chemicals pharmaceutically acceptable salt form include but not limited to multiple pharmaceutically acceptable salts forms such as acetate, hydrochlorate, sulfate, citrate, sulfonate, Salicylate.

High concentration phospholipid slow releasing preparation of the present invention, wherein bioactive ingredients content based on the gross weight meter of preparation, is about 0.01%～about 10% (g/g).

High concentration phospholipid slow releasing preparation of the present invention, wherein oil for injection content based on the gross weight meter of preparation, is about 10%～about 40% (g/g).

High concentration phospholipid slow releasing preparation of the present invention; Wherein alcoholic solution comprises dehydrated alcohol, ethanol-water solution, ethanol-normal saline solution, ethanol-phosphate-buffered liquor, ethanol-acetate salt buffer liquor, ethanol-carbonate buffer solution solution, ethanol-succinate buffer soln, ethanol-citrate buffer solution, ethanol-lactate buffer solution etc.; Wherein concentration of alcohol can be 80%～100% (v/v), and the content of this alcoholic solution in high concentration phospholipid slow releasing preparation is about 30%～about 5% (g/g).

One of the object of the invention provides a kind of high concentration phospholipid slow releasing preparation, comprises phospholipid, bioactive ingredients, alcoholic solution and/or oil for injection; Gross weight meter based on preparation; Wherein content of phospholipid is about 50%～about 85% (g/g), and bioactive ingredients content is about 0.01%～about 10% (g/g), and oil for injection content is about 10%～about 40% (g/g); Alcoholic solution content is about 30%～about 5% (g/g), and the concentration range of alcoholic solution is 80%～100% (v/v).

Of the present invention is that the in-situ injection phase change gel slow-released system of substrate is a kind of dosage form of novelty with phospholipid; Be that protein and peptide drugs or other drug are dissolved in the phospholipid-alcoholic solution of high concentration, become the liquid of good fluidity, after the injection body is interior; Ethanol spreads rapidly; The phospholipid of separating out forms gel, and packaging medicine has the good slow release effect.

One of another object of the present invention provides a kind of preparation in-situ injection phase change gel slow-released system, i.e. the method for high concentration phospholipid slow releasing preparation.

In one embodiment, method for preparing of the present invention comprises the steps:

(1) get bioactive ingredients and be dissolved in an amount of alcoholic solution, mixed with an amount of oil for injection again, form drug solution, the filtering with microporous membrane sterilization;

(2) under aseptic condition, get injection stage phospholipid and mix with drug solution, stirring is dissolved phospholipid fully, leaves standstill a moment to remove the bubble in the preparation, and packing seals, and promptly gets.

Because part of polypeptide less stable in liquid environment needs kept dry, so in another embodiment, the present invention also provides the method for preparing of another kind of high concentration phospholipid slow releasing preparation, comprises the steps:

(1) get an amount of bioactive ingredients and be dissolved in an amount of pure water or buffer salt solution, form drug solution, the filtering with microporous membrane sterilization, aseptic condition adds injection stage phospholipid, stirring and evenly mixing down;

(2) with above-mentioned semi-solid phospholipid gel packing, press the usual manner lyophilization, sealing;

(3) an amount of alcoholic solution is mixed with an amount of oil for injection, the filtering with microporous membrane sterilization, packing, sealing, subsequent use.

In concrete embodiment, the preferred 0.22 μ m of microporous filter membrane.

In concrete embodiment, preferably with the product of step (2) and (3), with the mode assembly packaging of test kit.

When product of the present invention uses, directly the oil for injection in the assembly packaging-ethanol mixture solution is injected lyophilizing phospholipid, firmly jolting or ultrasonic, dissolving promptly gets.

In-situ injection phase change gel slow-released system of the present invention can be used for drug administration by injection, also can be used for other form of medication such as topical administration.

In-situ injection phase change gel slow-released system of the present invention also can be used for repairing and expanding soft and/or sclerous tissues.

The present invention is through creationary research; With phospholipid, alcoholic solution and/or injection wet goods as primary raw material; Adopting protein and peptide drugs or micromolecule chemicals such as octreotide, erythropoietin, morphine sulfate etc. is model drug, and the mode through simple agitation is prepared into high concentration phospholipid slow releasing preparation.The gained preparation is a true solution, good fluidity, and easy drug administration by injection, experiment proves that it has the good slow release effect in the animal body.Examined or check the zest of high concentration phospholipid slow releasing preparation to the injection site, the result shows that said preparation has extraordinary biocompatibility, and can not produce medicine-feeding part stimulates.

Therefore, thereby the present invention has solved the problem that the vesicle type phospholipid gel of high phospholipid concentration is difficult for drug administration by injection well, has obtained slow release effect preferably again simultaneously, has a good application prospect.

 

Advantage of the present invention

High concentration phospholipid slow releasing preparation contains the alcoholic solution that can dissolve each other fully with water, be expelled in the body after, ethanol can be diffused into rapidly in the body fluid, the phospholipid in the preparation is separated out curing, becomes the carrier of medicament slow release, the release of control medicine.Experiment proof said preparation has the good slow release effect really in the animal body.High concentration phospholipid slow releasing preparation is a kind of true solution in fact, compare with the vesicle type phospholipid gel of identical phospholipid concentration have good fluidity, easy advantage such as drug administration by injection.In addition can also according to wrap the medicine carrying thing dissolubility select suitable liquid or liquid combination to come dissolved substance, to make the stabilization formulations of homogeneous transparent.In addition, the content of moisture-free or moisture is very low in the high concentration phospholipid slow releasing preparation, and contains ethanol in the preparation, can effectively suppress microbial growth, helps the stable and storage of preparation, thereby has enlarged the range of application of high concentration phospholipid slow releasing preparation.

 

Description of drawings

Below, specify embodiment of the present invention in conjunction with accompanying drawing, wherein:

Fig. 1 for HPLC-mass spectrography paint the linear regression curve of octreotide acetate.

Curve when Fig. 2 is the medicine of high concentration phospholipid slow releasing preparation of rat skin lower injection octreotide acetate.

Fig. 3 is the external standard curve of octreotide acetate.

 

The specific embodiment

Following examples are to further specify of the present invention, but never limit the scope of the present invention.Further set forth the present invention in detail with reference to embodiment below, but it will be appreciated by those skilled in the art that the present invention is not limited to the method for preparing of these embodiment and use.And those skilled in the art can be equal to replacement, combination, improvement or modification to the present invention according to description of the invention, but these all will comprise within the scope of the invention.

 

Embodiment 1

Get octreotide acetate 50mg, be dissolved in 85% (v/v) ethanol-pH4.0 lactic acid buffer solution of 1.5g, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add Ovum Gallus domesticus Flavus lecithin E80 7.0g, add the soybean oil 1.5g behind 0.22 μ m filtering with microporous membrane again, gnotobasis lower magnetic force stir about 0.5h; Dissolve fully to E80, obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill to the bubble complete obiteration; Packing, sealing promptly gets.

 

Embodiment 2

Get Thymopentin 20mg, be dissolved in the 2.0g dehydrated alcohol, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add Ovum Gallus domesticus Flavus lecithin E80 6.0g, add the medium chain length fatty acid triglyceride 2.0g of 0.22 μ m filtering with microporous membrane again, the about 0.5h of magnetic agitation; Dissolve fully to E80, obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill to the bubble complete obiteration; Packing seals, and promptly gets the high concentration phospholipid slow releasing preparation of Thymopentin.

 

Embodiment 3

Get insulin 100mg, be dissolved in 2.0g 90% (v/v) ethanol-normal saline solution, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add soybean phospholipid 5.5g, add the soybean oil 2.5g of 0.22 μ m filtering with microporous membrane again, the about 0.5h of magnetic agitation; Dissolve fully to soybean phospholipid, obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill to the bubble complete obiteration; Packing seals, and promptly gets the high concentration phospholipid slow releasing preparation of insulin.

 

Embodiment 4

Get erythropoietin (EPO) 30mg, be dissolved in 80% ethanol-phosphate buffer (pH value 6.9) of 2.0g, obtain drug solution; 0.22 μ m filtering with microporous membrane adds Ovum Gallus domesticus Flavus lecithin E80 5.0g, adds the Oleum Sesami of 3.0g 0.22 μ m filtering with microporous membrane again; The about 0.5h of magnetic agitation dissolves to E80 fully, obtains containing the milk yellow opaque liquid of great amount of bubbles; Leave standstill to the basic complete obiteration of bubble; Packing seals, and promptly gets the high concentration phospholipid slow releasing preparation of erythropoietin.

 

Embodiment 5

Get LEK 100 μ g, be dissolved in 85% (v/v) ethanol-water solution of 2.0g, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add hydrogenated soya phosphatide 7.0g, add the soybean oil of 1.0g 0.22 μ m filtering with microporous membrane again, the about 0.5h of magnetic agitation; Dissolve fully to hydrogenated soya phosphatide, obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill to the bubble complete obiteration; Packing seals, and promptly gets the high concentration phospholipid slow releasing preparation of LEK.

 

Embodiment 6

Get interleukins 16 mg, be dissolved in 80% (v/v) ethanol-water solution of 1.0g, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add Ovum Gallus domesticus Flavus lecithin E80 7.5g, add the medium chain length fatty acid triglyceride of 1.5g 0.22 μ m filtering with microporous membrane again, the about 0.5h of magnetic agitation; Dissolve fully to E80, obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill to the bubble complete obiteration; Packing seals, and promptly gets the high concentration phospholipid slow releasing preparation of interleukin.

 

Embodiment 7

Get interferon 30mg, be dissolved in 85% (v/v) ethanol-water solution of 1.5g, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add hydrogenated yolk lecithin 6.5g, add the soybean oil of 2.0g again, the about 0.5h of magnetic agitation through 0.22 μ m filtering with microporous membrane; Dissolve fully to hydrogenated yolk lecithin, obtain containing the opaque thick liquid of milk yellow of great amount of bubbles, leave standstill to the bubble complete obiteration; Packing seals, and promptly gets the high concentration phospholipid slow releasing preparation of interferon.

 

Embodiment 8

Get leuprorelin acetate 100mg, be dissolved in 1.5g 80% (v/v) ethanol-water solution, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add soybean phospholipid 7.5g, add the medium chain length fatty acid triglyceride of 1.0g again, the about 0.5h of magnetic agitation through 0.22 μ m filtering with microporous membrane; Dissolve fully to soybean phospholipid, obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill to the bubble complete obiteration; Packing seals, and promptly gets the high concentration phospholipid slow releasing preparation of leuprorelin acetate.

 

Embodiment 9

Get growth hormone 40mg, be dissolved in 95% (v/v) ethanol-water solution of 1.5g, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add dipalmitoyl phosphatidyl choline 6.0g, add the ethyl oleate of 2.5g through 0.22 μ m filtering with microporous membrane again, the about 0.5h of magnetic agitation is to dissolving fully; Obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill, obtain clear liquid to the bubble complete obiteration; Packing seals, and promptly gets the high concentration phospholipid slow releasing preparation of growth hormone.

 

Embodiment 10

Get acetic acid Ai Saina peptide 20mg and be dissolved in the 10ml water for injection, form drug solution, the sterilization of 0.22 μ m filtering with microporous membrane; Aseptic condition adds injection stage Ovum Gallus domesticus Flavus lecithin E80 3.0g down, and stirring and evenly mixing is pressed the packing of 2ml/ bottle;-40 ℃ freezing 4 hours, drained on the freeze dryer 24 hours, loose lyophilizing phospholipid; Sealing, subsequent use.

95% alcoholic solution and the 5g medium chain length fatty acid triglyceride of 5g is mixed, and the sterilization of 0.22 μ m filtering with microporous membrane is distributed into the 0.4g/ bottle, and sealing is subsequent use.

Face the time spent medium chain length fatty acid triglyceride-95% ethanol mixture solution is injected lyophilizing phospholipid, firmly jolting, dissolving promptly gets.

 

Embodiment 11

Take by weighing water soluble drug morphine sulfate 60mg, be dissolved in 2.0g 85% (v/v) ethanol-water solution, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add soybean phospholipid 6.0g, add the ethyl oleate of 2.0g through 0.22 μ m filtering with microporous membrane again, magnetic agitation to soybean phospholipid dissolves fully; Obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill, obtain clear liquid to the bubble complete obiteration; Packing, sealing promptly gets.

 

Embodiment 12

Take by weighing fat-soluble medicine nimodipine 40mg, be dissolved in the 2.0g dehydrated alcohol, obtain drug solution, 0.22 μ m filtering with microporous membrane; Add Ovum Gallus domesticus Flavus lecithin E80 7.0g, add the medium chain length fatty acid triglyceride of 1.0g through 0.22 μ m filtering with microporous membrane again, magnetic agitation is dissolved to E80 fully; Obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill, obtain clear liquid to the basic complete obiteration of bubble; Packing, sealing promptly gets.

?

The selection of embodiment 13 concentration of alcohol

Take by weighing 4 parts of Ovum Gallus domesticus Flavus lecithins (E80), every part of 7.0g adds Different concentrations of alcohol respectively to 10.0g, stirs 2 hours, observes outward appearance, and measures viscosity, and the result sees table 1.It is thus clear that concentration of alcohol obtains the solution of good fluidity dissolving phospholipid well more than 80%.

The screening of table 1 concentration of alcohol

Concentration of alcohol	100	95	90	85	80	75
							Outward appearance	Clear	Clear	Clear	Clear	Clear	Muddiness is not clarified
Viscosity	152.7	158.1	156.8	161.2	173.6	326.3

Embodiment 14 different types of host materials are to the influence of preparation viscosity

Take by weighing different types of phospholipid 7.0g, oily 1.5g and alcoholic solution 1.5g respectively by table 2 prescription, stirred 2 hours, measure viscosity, the result sees table 2.It is thus clear that it is little to the viscosity influence of preparation to add different types of phospholipid, oil and alcoholic solution.

The different types of host material of table 2 is to the influence of preparation viscosity

Prescription	Phospholipid	Oil	Alcoholic solution	Viscosity
					Prescription 1	Soybean phospholipid	Semen Maydis oil	Dehydrated alcohol	468.5
Prescription 2	Soybean phospholipid	Oleum Sesami	90% ethanol-water solution	471.4
					Prescription 3	Egg yolk lecithin	Oleum Camelliae	85% ethanol-normal saline solution	460.2
Prescription 4	Egg yolk lecithin	Fish oil	95% ethanol-pH7.4 phosphate-buffered liquor	469.3
					Prescription 5	Hydrogenated soya phosphatide	Oleum Gossypii semen	90% ethanol-pH4.0 lactate buffer solution	477.6
Prescription 6	Hydrogenated soya phosphatide	Medium chain length fatty acid triglyceride	80% ethanol-pH6.0 citrate buffer solution	480.9
					Prescription 7	Dipalmitoyl phosphatidyl choline	Ethyl oleate	85% ethanol-pH5.0 succinate buffer soln	475.7
Prescription 8	Dipalmitoyl phosphatidyl choline	Soybean oil	95% ethanol-pH4.6 acetate salt buffer liquor	462.6

The screening of embodiment 15 consumption of ethanol

Take by weighing the soybean oil and the ethanol solution of injection Ovum Gallus domesticus Flavus lecithin (E80), filtration sterilization respectively by table 3 prescription, aseptic condition mixes down, is stirred to dissolving, packing, sealing.Carry out the local irritation experiment by hereinafter 3 methods of testing, tentatively investigate the zest of different dehydrated alcohol consumptions the injection site.

Experimental technique: with experiment 3.Inserting needle position and medicine are observed its variation in subcutaneous position under the injection end postscript.The result sees table 3.It is thus clear that the content of dehydrated alcohol is lower than at 30% o'clock in prescription, preparation does not have zest or zest very little to medicine-feeding part, and reversible.In theory, the alcoholic solution that contains a certain amount of moisture replaces dehydrated alcohol, and its toxicity should be lower.

The screening of table 3 consumption of ethanol

Prescription	Phospholipid E80 (g)	Oil (g)	Dehydrated alcohol (g)	The local irritation experimental result
					Prescription 1	5.0	4.0	1.0	Medicine-feeding part does not have tangible zest
Prescription
	2	5.0	3.0	2.0	Medicine-feeding part does not have tangible zest
Prescription 3						5.0	2.5	2.5	Medicine-feeding part does not have tangible zest
	Prescription
4		5.0	2.0	3.0	Redness is arranged slightly, but disappear after 2-3 days
	Prescription 5					5.0	1.0	4.0	Medicine-feeding part is red and swollen, and skin begins to fester after 5-6 days

[effect experiment]

The viscosity of [experiment 1] high concentration phospholipid slow releasing preparation reaches the comparison with vesicle type phospholipid gel preparation

Find that through experiment though the concentration of phospholipid is very high in the high concentration phospholipid slow releasing preparation, viscosity is within injectable scope, and is more much lower than vesicle type phospholipid gel preparation identical even lower phospholipid concentration.

Preparation reference literature method (the M. Brandl of vesicle type phospholipid gel preparation; Liposome Technol. 1 (1993) 49 – 65); Take by weighing Ovum Gallus domesticus Flavus lecithin E80 3.0g, 3.5g, 4.0g, 4.5g, 5.0g respectively, add water for injection respectively, be stirred to mixing to 10.0g; High pressure homogenize promptly gets again.

The viscosity of the vesicle type phospholipid gel preparation of high concentration phospholipid slow releasing preparation that comparative example 1,2,4,5,8,10 makes and several kinds of different phosphate lipid concentrations.Be experimental result below:

The comparison of table 4 high concentration phospholipid slow releasing preparation and vesicle type phospholipid gel preparation viscosity

Annotate: above viscosity number is under 25 ℃ of conditions and records.

Can find out that from above experimental result for high concentration phospholipid slow releasing preparation, its viscosity increases with the rising of content of phospholipid in the preparation, but the amplitude that changes is less, changes little generally.And for vesicle type phospholipid gel preparation, its viscosity sharply increases with the rising of content of phospholipid in the preparation, and its viscosity has reached 11659.4cP when phospholipid concentration reaches 40%, and injection is difficulty quite, and when phospholipid concentration reaches 50%, does not almost have flowability.And find that further the viscosity of high concentration phospholipid slow releasing preparation of the present invention is less than the viscosity of the vesicle type phospholipid gel preparation of equal content of phospholipid, therefore, high concentration phospholipid slow releasing preparation of the present invention has better syringeability.

 

The slow release characteristic of [experiment 2] high concentration phospholipid slow releasing preparation

Adopt the slow release effect of experiment exam embodiment 1 in the animal body.

It is subcutaneous that the product of embodiment 1 (phospholipid concentration is 70%) is expelled to male Wistar rat, regularly gets blood, adopts the octreotide acetate content in HPLC-mass spectrometric determination blood plasma, examines or check its slow releasing function time.

1. the preparation of standard solution

Precision takes by weighing 13.18 mg octreotide acetate standard substance, puts in the 10 ml volumetric flasks, is dissolved in water, and is settled to scale, is mixed with the storing solution of 1.318 mg/ml.It is an amount of to get this storing solution, uses methanol-water (1:1 v/v) to be diluted to 0,6.59,13.18,26.36,65.90,105.44 and 131.80 ng/ml respectively, obtains a series of octreotide acetate standard solution.

2. the preparation of standard curve

Get blank plasma 100 μ l, add octreotide acetate standard serial solution 25 μ l, be mixed with and be equivalent to the sample that plasma drug level is respectively 0,0.51,1.01,2.03,5.07,8.11 and 10.14 ng/ml; Mixing is placed in 37 ℃ of shaking tables and hatches 30min; Hatch and finish back adding 200 μ l acetonitriles, whirlpool mixing 5min, centrifugal (13500rpm) 5min; Supernatant is crossed 0.22 μ m filtering with microporous membrane, get the subsequent filtrate sample introduction.With known drug concentration in the blood plasma is abscissa, and peak area is a vertical coordinate, and drawing standard curve, gained linear regression equation are y=53.973x+0.5335 (R2=0.9998), is standard curve (seeing accompanying drawing 1).The range of linearity that this method is measured octreotide acetate in the rat plasma sample is 0.51-10.14ng/ml.

3. the processing of plasma sample

Get 100 μ l plasma samples, add 25 μ l methanol-waters (1:1 v/v) earlier, add 200 μ l acetonitriles again, whirlpool mixing 5min, centrifugal (13500rpm) 5min crosses 0.22 μ m filtering with microporous membrane with supernatant, gets the subsequent filtrate sample introduction.

4. chromatograph and mass spectrum condition

The chromatographic condition chromatographic column is a Zorbax SB-Aq post, 5 μ m particle diameters, 150 * 4.6 mmI.D., U.S. Anjielun Technology Co., Ltd; Mobile phase is acetonitrile: 0.023% aqueous formic acid=25:75, and, isocratic elution; Flow velocity is 0.3 ml/min; Column temperature is 30 ℃; Sample size is 1 μ l.

Mass spectrum condition ion source: ESI electro-spray ionization source; Dwell:250ms; Fragmentor 200V; Collision Energy:40V; Delta Emv (+): 100V; Gas Temperature:350 ℃; Gas Flow:8L/min; Nebulizer:30psi.The cation mode detects, and scan mode is multiple-reaction monitoring (MRM).The ionic reaction that is used for Mass Spectrometer Method is respectively m/z 510 → m/z 120 and m/z 510 → m/z 159, and wherein m/z 510 → m/z 120 is used for quantitative analysis, and 510 → m/z 159 is used for qualitative analysis.

5. interior animal experiment

Get 6 male Wistar rats, body weight 200 ± 20g, Sichuan University's West China medical experiment animal center provides.Fasting 12 h before the administration press the sample of dosage subcutaneous injection embodiment 1 preparation of 20mg/kg, ad lib and drinking-water after the administration.0.5h after (0h) and the administration, 1h, 2h, 4h, 6h, 9h, 12h, 24h, 36h, 48h, 72h, equi-time point docking in 10 days, 20 days, 30 days are got blood 300 μ l in heparinization EP pipe before the administration; Centrifugal (4500rpm) 5min; Separated plasma is preserved to be measured in-20 ℃ of refrigerators.After above-mentioned plasma sample disposal methods, the LC-MS assay determination.

Measure the result: the rat skin lower injection administration can also measure medicine (curve is seen accompanying drawing 2 during medicine) after 30 days, and visible high concentration phospholipid slow releasing preparation has fabulous slow release effect.

 

The zest research at [experiment 3] high concentration phospholipid slow releasing preparation drug administration by injection position

Product with embodiment 1 is the zest of representative examination high concentration phospholipid slow releasing preparation to medicine-feeding part.Carry out the local irritation experiment with reference to " chemicals zest, anaphylaxis and hemolytic investigative technique guideline ", and with only drug, phospholipid and alcoholic acid Comparative formulation product are compared.

The preparation of Comparative formulation: get octreotide acetate 50mg, be dissolved in 85% (v/v) ethanol-water solution of 3.0g, obtain drug solution; 0.22 μ m filtering with microporous membrane adds Ovum Gallus domesticus Flavus lecithin E80 7.0g, gnotobasis lower magnetic force stir about 0.5h; Dissolve fully to E80, obtain containing the milk yellow opaque liquid of great amount of bubbles, leave standstill to bubble and disappear basically; Packing, sealing promptly gets.

 

1. test method

Get 8 rabbit, be divided into two groups, each 4, male and female half and half.If consubstantiality left and right sides self matching type is adopted in the normal saline contrast.Test preceding 24 hours to the processing (earlier with shears mao the cutting to suitable length of district of will lose hair or feathers, the reuse depilatory is further handled) of losing hair or feathers of the administration district of both sides, back.The depilation scope is 3cm * 3cm each side.Whether the inspection skin of unhairing has the skin of damage to make an experiment because of the depilation damaged before the administration.

Adopt the subcutaneous injection administration, adjust the inserting needle position, it is subcutaneous to make medicine just in time be expelled to the depilation position.The product of 4 rabbit one side injection embodiment 1, the opposite side injecting normal saline.The product of other 4 rabbit one side injection Comparative Examples, the opposite side injecting normal saline.Injection finishes under the postscript inserting needle position and medicine in subcutaneous position, so that observation and carry out the medicine-feeding part histopathological examination.In 72 hours animal and injection site are carried out perusal after the administration, the observation period finishes the back and randomly draws two rabbit for every group and carry out the medicine-feeding part histopathological examination.The rabbit that stays continues to observe, and carries out histopathological examination after 14 days, to understand the degree of reversibility of irritative response.

2. result of the test

In 72 hours animal and injection site are carried out perusal after the administration, just begun to find to compare with a side of injecting normal saline, a side of injection high concentration phospholipid slow releasing preparation is heaved a little a little, but other abnormal phenomenas such as erythema or edema do not occur.As time goes on, heave gradually and diminish, do not seen obvious difference by 72 hours.72 hours medicine-feeding part histopathological examination result shows that the tissue of injection high concentration phospholipid slow releasing preparation one side is as broad as long basically with organizing of injecting normal saline one side, does not all have tangible medicine-feeding part zest.

The zest phenomenon does not appear in two rabbit that stay medicine-feeding part during 72h after the administration ~ 14 day, and the position of As time goes on losing hair or feathers grows hair gradually and comes.14 days histopathological examination result shows that injection high concentration phospholipid slow releasing preparation is the same with injecting normal saline, tangible medicine-feeding part zest all do not occur.

Above result of the test shows that high concentration phospholipid slow releasing preparation can not produce zest to medicine-feeding part, has extraordinary biocompatibility.

And the administration of Comparative formulation group is after 4 hours, and the injection site occurs red and swollen, and after 5 days fester in red and swollen position, shows tangible local toxicity.

Claims (10)

1. pharmaceutical composition that comprises phospholipid, alcoholic solution and/or oil for injection.

2. pharmaceutical composition according to claim 1 is characterized in that being in-situ injection phase change gel slow releasing preparation.

3. the described a kind of in-situ injection phase change gel slow releasing preparation of claim 2; It is characterized in that comprising phospholipid, bioactive ingredients, alcoholic solution and/or oil for injection; Based on the gross weight meter of preparation, wherein content of phospholipid is about 50%～about 85% (g/g), and bioactive ingredients content is about 0.01%～about 10% (g/g); Oil for injection content is about 10%～about 40% (g/g), and alcoholic solution content is about 30%～about 5% (g/g).

4. the described a kind of in-situ injection phase change gel slow releasing preparation of claim 3, the concentration range that it is characterized in that alcoholic solution is about 80%～100% (v/v).

5. pharmaceutical composition according to claim 2 is characterized in that said phospholipid comprises, one or more combination in natural phospholipid, semi-synthetic phospholipid, the synthetic phospholipid.

6. pharmaceutical composition according to claim 2 is characterized in that said oil for injection comprises, the combination of soybean oil, Semen Maydis oil, Oleum Sesami, Oleum Camelliae, fish oil, medium chain length fatty acid triglyceride, Oleum Gossypii semen, ethyl oleate and above-mentioned substance.

7. pharmaceutical composition according to claim 2; It is characterized in that said alcoholic solution comprises dehydrated alcohol, ethanol-water solution, ethanol-normal saline solution, ethanol-phosphate-buffered liquor, ethanol-acetate salt buffer liquor, ethanol-carbonate buffer solution solution, ethanol-succinate buffer soln, ethanol-citrate buffer solution, ethanol-lactate buffer solution etc.; Wherein concentration of alcohol can be about 80%～100% (v/v), and the content that this alcoholic solution is injected in the phase change gel slow releasing preparation in position is about 30%～about 5% (g/g).

8. method for preparing the described in-situ injection phase change gel of claim 2 slow releasing preparation is characterized in that may further comprise the steps:

(1) get bioactive ingredients and be dissolved in an amount of alcoholic solution, mixed with an amount of oil for injection again, form drug solution, the filtering with microporous membrane sterilization;

(2) under aseptic condition, get injection stage phospholipid and mix with drug solution, stirring is dissolved phospholipid fully, leaves standstill a moment to remove the bubble in the preparation, and packing seals, and promptly gets.

9. method for preparing the test kit of the described in-situ injection phase change gel of claim 2 slow releasing preparation is characterized in that may further comprise the steps:

(1) get an amount of bioactive ingredients and be dissolved in an amount of pure water or buffer salt solution, form drug solution, the filtering with microporous membrane sterilization, aseptic condition adds injection stage phospholipid, stirring and evenly mixing down;

(2) with above-mentioned semi-solid phospholipid gel packing, press the usual manner lyophilization, sealing;

(3) an amount of alcoholic solution is mixed with an amount of oil for injection, the filtering with microporous membrane sterilization, packing, sealing, subsequent use;

(4) product of step (2) and (3) is with the mode assembly packaging of test kit.

10. the application of the compositions of claim 1 in the in-situ injection phase change gel slow releasing preparation of preparation reduction ethanol side effect.

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