CN102526753B - In-situ phase change gel slow release system taking phospholipid as substrate and preparation method thereof - Google Patents
In-situ phase change gel slow release system taking phospholipid as substrate and preparation method thereof Download PDFInfo
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- CN102526753B CN102526753B CN201110420104.4A CN201110420104A CN102526753B CN 102526753 B CN102526753 B CN 102526753B CN 201110420104 A CN201110420104 A CN 201110420104A CN 102526753 B CN102526753 B CN 102526753B
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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
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 with alcoholic solution and/or the oil for injection of phospholipid, variable concentrations, is medical technical field.
Background technology
Along with the rise and development of biotechnology, increasing protein and peptide drugs is developed and for diagnosis and the treatment of disease.Compare with micromolecule chemicals, in protein and peptide drugs and body, normal physiologically substance is more approaching, has pharmacologically active high, and toxic and side effects is low, and dosage is little, determined curative effect, the advantages such as high specificity.But also exist a lot of problems: on the one hand, protein and peptide drugs molecular mass is large, and poor stability, easily by the Proteolytic enzyme enzymatic degradation in gastrointestinal tract, therefore, can not adopt common gastrointestinal administration mode conventionally, can only pass through drug administration by injection simultaneously; On the other hand, the biological half-life of this class medicine is shorter, is decomposed soon by body, and the curative effect that needs lasting injection or intravenous drip to guarantee medicine, has increased the weight of patient's health, psychology and financial burden.
Therefore, people wish to develop a kind of Atrigel of injectable protein and peptide drugs.
At present, the Injectable sustained release drug-supplying system having gone on the market mainly comprises PLGA (Poly(D,L-lactide-co-glycolide) microsphere and multivesicular liposome (MVL).Microsphere refers to medicine dissolution or is dispersed in the small spherical entity forming in macromolecular material substrate, belongs to matrix type skeleton microgranule.PLGA is the copolymer being 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 successfully kind.Triptorelin is one of p-GLU-HIS-TRP-SER-TYR-D-TRP-LEU-ARG-PRO-GLY-NH2, and its PLGA microsphere is developed by French Ipsen company, and listing in 1986 can slow release 1 month, is first polypeptide microspheres product.Alarelin is the first class national new drug of Shanghai Li Zhu 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 monthly medication 1 time, animal vivo test proves that its slow release effect is reliable.
Although PLGA microsphere has good slow-releasing and controlled-releasing action and biocompatibility, but due to its complicated process of preparation, drug loading is lower, the organic solvent using in preparation process has residual in preparation, and in degradation process, produce lactic acid and hydroxyacetic acid can cause the decline of injection site pH value, thereby produce local aseptic inflammation.Above shortcoming and defect has limited the application of PLGA microsphere greatly.
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 by lipoid and immobilized artificial membrane, forms a lot of vesicles.In this discontinuous vesicle, when certain vesicle breaks, medicine only disengages from the vesicle breaking, and complete vesicle still maintains the original state, thereby has good slow release effect.The height that traditional liposomal is generally difficult to reach water soluble drug with prior art is sealed and low seepage, multivesicular liposome has the advantages such as envelop rate is high, drug leakage is 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 product introduction research and development and clinical research stage.
Although multivesicular liposome route of administration is various, there is good slow releasing function and bank effect, it also has many weak points: first, wish is prepared MVL, at least needs to contain parents' lipid (as phospholipid) and neutral lipid (as triglyceride).If form traditional unilamelar liposome (ULV) or multilamelar liposome (MLV) without neutral lipid.Secondly, because water inside and outside MVL exists drug level poor, the leakage problems of small-molecule drug still exists, and phospholipid bilayer also there will be partial rupture when storing, and its sedimentation and rendezvous problem of occurring at storage process are not yet resolved, may affect the stability of preparation.The 3rd, the MVL product of exploitation is suspension at present, is not easy to storage and transport.The 4th, the preparation process of MVL is complicated, very harsh to working condition requirement, is difficult for expanding production.In addition, the medication amount that MVL bag carries is limited, and this also makes its development be subject to certain restrictions.
In addition, the vesicle type phospholipid gel (vesicular phospholipid gels, VPG) in addition of studying.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.The in the situation that of water as solvent, in vesicle and all contain the water of certain volume between vesicle, can loaded with water-soluble, fat-soluble and amphipathic medicine.For water soluble drug, the inside and outside intimate equal water volume of vesicle not only makes its envelop rate improve, and medicine in vesicle and vesicle all exist outward, interior extracellular concentration reaches unanimity, and has weakened to a certain extent the impact on stability such as the drug leakage that causes due to Concentraton gradient.
Now existing more document has disclosed the report as peptide medicament Atrigel about vesicle type phospholipid gel.For example, Brandl, the research group at the people places such as Winter adopts respectively micromolecule chemicals calcein and 5-fluorouracil, micromolecule polypeptide cetrorelix and usings high molecular weight protein erythropoietin as model drug, in homemade release flow cell, release in vitro behavior (C. Tardi, Journal of Controlled Release 55 (1998) 261 –s 270 of VPG during as drug depot are usingd in research; 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, VPG is better to the slow release effect of high molecular weight protein, can reach hundreds of hours; Patent application 20110036587.8 is found by research in the body of animal, concentration also has longer 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, likely causes toxic and side effects.Along with the increase of phospholipid concentration, prominent releasing reduces.When phospholipid concentration is elevated to 40%, although slow release effect is better, its viscosity reaches 14.3PaS(W. Tian, Journal of Controlled Release 142 (2010) 319 – 325), quite thickness, almost can not inject.So can find out from existing research, vesicle type phospholipid gel is not too suitable for as injection polypeptide slow-released carrier.
Patent documentation 93119112.2 has disclosed a kind of lecithin gel that forms in vivo with the Pharmaceutical composition of sustained release bioactive compound, comprises lecithin, pharmaceutically acceptable for intramuscular or subcutaneous injection and water-fast lecithin solvent and bioactive compound substantially.Briefly, this Pharmaceutical composition is exactly lecithin organic solution or the suspension that contains bioactive compound.By intramuscular or this solution of subcutaneous injection or suspension, can form in vivo lecithin gel, by absorbing moisture body fluid between the moisture gap in injection site, form in vivo lecithin gel.According to patent description, medicine length of release time from compositions is decided by the composition of said composition.In addition can also control by adding excipient.Yet, in the main component due to said composition, lack polar solvent, in said composition, there is certain difficulty in the medicinal application that water soluble drug or polarity are stronger.Although say in document and active component first can be dissolved in a small amount of water or be applicable in the buffer solution of this concrete active component, and then water or buffer are added in 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, during as local drug delivery for lasting, it is 10-90% that said composition limits phospholipid fraction concentration in certain embodiments, maximum quantity is 45% at the most in certain embodiments, but do not clearly state, is which embodiment.Compositions prepared in embodiment 1 is because the dissolubility of phospholipid in selected solvent is not high, in order to be injected into smoothly human body under suitable viscosity, phospholipid concentration is all lower, and the large usage quantity of propylene glycol, glycerol equal solvent, therefore, even if said composition is injected human body on a small quantity, also can cause that local swelling even festers.In addition, its process is: weigh up and mixing soya beans lecithin (Phospholipon 90G in clean container, the injectable grade soybean lecithin that comprises approximately 90% phosphatidylcholine, Phospholipid GmbH), medium chain triglyceride (Miglyol812, Sasol Corp.), sucrose and propylene glycol, add dehydrated alcohol, dissolve all solids to form limpid yellow solution.Evacuation is to remove ethanol, until residue ethanol content is less than 5% of gross weight.Heat this mixture to 60 ℃ to form clear solution, then by sterilization filter, filter this solution.By filtrate cool to room temperature, obtain yellow transparent gel uniformly, dissolve as can be seen here the not ethanol of above-mentioned phospholipid but propylene glycol.
More crucially, this application is not carried out experiment in body, and it can reach the slow release effect of expection there is no 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 its slow releasing function is very poor completely.
Therefore, develop a kind of can being expelled in body and reach good slow release effect, viscosity phospholipid slow releasing preparation lower, drug administration by injection smoothly just becomes a difficult problem and the focus that this area is studied again.
Summary of the invention
One of object of the present 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 when the content of phospholipid in ethanol is up to 85%(g/g), still clear, can flow.According to this character, inventor's imagination, by medicine dissolution in the alcoholic solution of this high concentration phospholipid, after being injected in vivo, because second alcohol and water can dissolve each other completely, after ethanol is diffused into body fluid rapidly, 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 is also completely different from the common phospholipid gel of mentioning in prior art.Patent documentation 93119112.2 has disclosed lecithin organic solution or the suspension that contains bioactive compound, this organic solution is substantially water insoluble, after intramuscular or subcutaneous injection, then form in vivo lecithin gel by absorbing moisture body fluid between the moisture gap in injection site.
According to above-mentioned unexpected discovery, inventor be take octreotide acetate thus as model drug, has prepared the alcoholic solution containing 70% phospholipid, gives after rat skin lower injection 1ml, and medicine can steadily discharge almost about one month.But adopt single ethanol as solvent, side effect is also extremely obvious, there is red and swollen and the phenomenon of festering in most of rat injection site.This is also in prior art, to be avoided using a large amount of ethanol as one of major reason of injection solvent as far as possible.
In order to overcome the side effect of ethanol, by creationary research, we are surprised to find, if replaced part ethanol with soybean oil in preparation, no longer there is redness and the phenomenon of festering in rat injection site, this is the effective solution that is conducive to the industrialization of this slow releasing preparation system.
One of object of the present invention, is to provide a kind of compositions that comprises phospholipid, alcoholic solution and/or oil for injection.
By more deep research, inventor finds, adds oil for injection in solvent, is the consumption that reduces ethanol on the one hand; On the other hand, reduce the diffusion velocity of ethanol, thereby not only realized effective utilization of phospholipid gel slow-released system, also reduced widely the side effect of ethanol.
One of object of the present invention, is to provide and a kind ofly with phospholipid, alcoholic solution and/or oil for injection, is prepared into high concentration phospholipid slow releasing preparation, be i.e. in-situ injection phase change gel slow-released system of the present invention or in-situ injection phase change gel slow releasing preparation.
Known 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 as soybean oil, replace part ethanol, reduce the side effect of ethanol, finally realized thus the present invention.
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.
Although have the technical scheme that comprises lecithin, oil for injection, ethanol, active component in prior art, the concrete preparation of these compositionss is generally Emulsion, the mechanism of action is different from original position phase change gel preparation of the present invention completely.
One of object of the present invention, is to provide a kind of compositions that comprises active constituents of medicine, phospholipid, alcoholic solution and/or oil for injection.After said composition is injected in vivo, ethanol spreads rapidly, and the phospholipid of separating out forms gel, and packaging medicine active component, has good slow releasing function.
One of object of the present invention, is to provide 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 object of the present 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 being injected in vivo, ethanol spreads rapidly, and the phospholipid of separating out forms gel, packaging medicine, has good slow releasing function.
Term herein " in-situ injection phase change gel slow-released system ", also comprises high concentration phospholipid slow releasing preparation.
The phospholipid that is applicable to high concentration phospholipid slow releasing preparation of the present invention, includes but not limited to: one or more combination in natural phospholipid, semi-synthetic phospholipid, synthetic phospholipid.
Described natural phospholipid, includes but not limited to Ovum Gallus domesticus Flavus lecithin, soybean phospholipid or its combination etc.
Described semi-synthetic phospholipid, includes but not limited to hydrogenated yolk lecithin, hydrogenated soya phosphatide or its combination etc.
Described synthetic phospholipid, includes but not limited to one or more the combination etc. in DPPE, dipalmitoyl phosphatidyl choline, distearoyl phosphatidylcholine, dimyristoyl phosphatidyl choline, DOPE, DPPG, DPPA.
In specific embodiment, the preferred Ovum Gallus domesticus Flavus lecithin E80 of the present invention.
In specific embodiment, high concentration phospholipid slow releasing preparation of the present invention, the gross weight meter based on preparation, wherein content of phospholipid, is approximately 50%～about 85%(g/g).
Be applicable to prepare the bioactive ingredients of high concentration phospholipid slow releasing preparation of the present invention, it 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.
Being applicable to medicine of the present invention includes but not limited to as follows:
Protein and peptide, includes but not limited to, insulin, glucagon, alpha-interferon, beta-interferon, interleukin, erythropoietin (EPO), promoting sexual gland hormone, granulocyte-macrophage colony stimutaing factor (GM-GSF), tissue-type plasminogen activator, streptokinase, cell growth factor, thrombin, prostaglandin, antithrombase, hirudin, histrelin, growth hormone-releasing peptide-2, His-D-Trp-Ala-Trp-D-Phe-Lys-NH2, cetrorelix, bivalirudin, arginine vasopressin, Schweine-Vasopressin, Desmopressin, gonadorelin, leuprorelin, De She Rayleigh, buserelin, triptorelin, goserelin, alarelin, Sermorelin, nafarelin, lutrelin, fertirelin, Hexarelin, octreotide, Exenatide, human glucagon-like-peptide-1, growth hormone, somatostatin, ziconotide, thymosin a1, sincalide, Thymopentin, 1: PN: WO02056903 PAGE: 25 claimed protein, aviptadil, leucine enkephalin, 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, Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2, neurotensin, human brain natriuretic peptide, atrial natriuretic peptide, hypertensinⅠ, angiotensin II, angiotensin II I, sleep-inducing peptide, hypermnesia peptide, dynorphin, endorphins, angiotensin, enfuirtide, CJC-1295, elcatonin, Thymosin β4, vassopressin, Pitressin Tannate, felypressin, ornipressin, melanotan II, spleen pentapeptide, Pramlintide, vapreotide, thyroliberin, silk relies short cortex 18 peptides, sweet essence is urged cortex 18 peptides, zinc is urged cortex 24 peptides, short cortex 24 peptides, short cortex 25 peptides, short cortex 28 peptides, vasoactive intestinal peptide, brain natriuretic factor(peptide), Kallidin I, orphanin, asparaginase etc., micromolecule chemicals class, include 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, receive many Nores, indole Nore, Ah 's Nore, Mei Tuonuoer, A Pu Nore, vinegar fourth Nore, nifedipine, verapamil, diltiazem, spectrum Ni Laming, piperazine can former times woods, bepridil, nicorandil, molsidomine, dilazep, trimetazidine, quinidine sulfate, the 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, receive many Nores, indole Nore, Ah 's Nore, Mei Tuonuoer, A Pu Nore, vinegar fourth Nore, peso Nore, his Nore doubly, prazosin, terazosin, doxazosin, trimazosin, draw 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, diethylstilbestrol propionic ester, diethylstilbestrol phosphate ester, estradiol cypionate, Dai, medroxyprogesterone acetate, megestrol acetate, chlormadinone acetate, 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, norpregnenes, nomegestrol, trimegestone, diphenhydramine, promethazine, tripelennamine, chlorphenamine, buclizine, phenindamine, Cyproheptadine, clemastine, azelastine, astemizole, loratadine, cetirizine, mizolastine, teldane,
fexofenadine fourth, Efletirizine, Desloratadine, ebastine,
promethazine, pheniramine, brompheniramine, tolpropamine, pyrrobutamine, the pyridine of music score of Chinese operas profit, 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 is (as 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, Lenalidomide, aspirin, magnesium salicylate, choline salicylate, salicylamide, salsalate, diflunisal, indomethacin (indometacin), sulindac, acetaminophen (acetaminophen), aminophenazone, oxyphenbutazone, Phenylbutazone, mefenamic acid, clofenamic acid, diclofenac, ibuprofen, naproxen, 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 the multiple pharmaceutically acceptable salts forms such as acetate, hydrochlorate, sulfate, citrate, sulfonate, Salicylate.
High concentration phospholipid slow releasing preparation of the present invention, bioactive ingredients content wherein, the gross weight meter based on preparation, is approximately 0.01%～about 10%(g/g).
High concentration phospholipid slow releasing preparation of the present invention, oil for injection content wherein, the gross weight meter based on preparation, is approximately 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), the content of this alcoholic solution in high concentration phospholipid slow releasing preparation is approximately 30%～about 5%(g/g).
One of object of the present invention, a kind of high concentration phospholipid slow releasing preparation is provided, comprise phospholipid, bioactive ingredients, alcoholic solution and/or oil for injection, gross weight meter based on preparation, wherein content of phospholipid is approximately 50%～about 85%(g/g), bioactive ingredients content is approximately 0.01%～about 10%(g/g), oil for injection content is approximately 10%～about 40%(g/g), alcoholic solution content is approximately 30%～about 5%(g/g), and the concentration range of alcoholic solution is 80%～100%(v/v).
It is of the present invention that to take the in-situ injection phase change gel slow-released system that phospholipid is substrate be a kind of dosage form of novelty, 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 being injected in vivo, ethanol spreads rapidly, the phospholipid of separating out forms gel, and packaging medicine, has good slow releasing function.
One of another object of the present invention, provides a kind of in-situ injection phase change gel slow-released system, i.e. method of high concentration phospholipid slow releasing preparation prepared.
In one embodiment, preparation method of the present invention, comprises the steps:
(1) get bioactive ingredients and be dissolved in appropriate alcoholic solution, more mixed with appropriate oil for injection, form drug solution, filtering with microporous membrane sterilizing;
(2) under aseptic condition, get injection stage phospholipid and mix with drug solution, stir phospholipid is dissolved completely, standing a moment, packing, sealed, and obtains to remove the bubble in preparation.
Due to part of polypeptide less stable in liquid environment, need kept dry, therefore in another embodiment, the present invention also provides the preparation method of another kind of high concentration phospholipid slow releasing preparation, comprises the steps:
(1) get appropriate bioactive ingredients and be dissolved in appropriate pure water or buffer salt solution, form drug solution, filtering with microporous membrane sterilizing, adds injection stage phospholipid, stirring and evenly mixing under aseptic condition;
(2) by above-mentioned semi-solid phospholipid gel packing, lyophilization in the usual way, sealing;
(3) appropriate alcoholic solution and appropriate oil for injection is mixed, filtering with microporous membrane sterilizing, packing, sealing, standby.
In specific embodiment, microporous filter membrane is 0.22 μ m preferably.
In specific embodiment, preferably by the product of step (2) and (3), with the mode assembly packaging of test kit.
When product of the present invention is used, directly oil for injection-ethanol mixture the solution in assembly packaging is injected to lyophilizing phospholipid, firmly jolting or ultrasonic, dissolves, and obtains.
In-situ injection phase change gel slow-released system of the present invention can be for drug administration by injection, also can be for other form of medication such as topical administrations.
In-situ injection phase change gel slow-released system of the present invention also can and expand soft and/or sclerous tissues for reparation.
The present invention is by creationary research, using phospholipid, alcoholic solution and/or injection wet goods as primary raw material, adopt protein and peptide drugs or micromolecule chemicals as octreotide, erythropoietin, morphine sulfate etc. be model drug, the mode by simple agitation is prepared into high concentration phospholipid slow releasing preparation.Gained preparation is true solution, good fluidity, and easily drug administration by injection, experimental results show that in animal body that it has good slow release effect.Examined or check the zest of high concentration phospholipid slow releasing preparation to injection site, result shows that said preparation has extraordinary biocompatibility, can medicine-feeding part not produced and be stimulated.
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 again good slow release effect 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 completely with water, and after being expelled in body, ethanol can be diffused into rapidly in body fluid, and the phospholipid in preparation is separated out curing, becomes the carrier of medicament slow release, controls the release of medicine.In animal body, experimental results show that said preparation has good slow release effect really.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 there is good fluidity, the easy advantage such as drug administration by injection.Can also select suitable liquid or liquid combination to carry out dissolved substance according to the dissolubility that will wrap medicine carrying thing in addition, to make the stabilization formulations of homogeneous transparent.In addition, in high concentration phospholipid slow releasing preparation, the content of moisture-free or moisture is very low, and in preparation, contains ethanol, can effectively suppress microbial growth, is conducive to the stable and storage of preparation, thereby has expanded the range of application of high concentration phospholipid slow releasing preparation.
Accompanying drawing explanation
Below, describe by reference to the accompanying drawings embodiment of the present invention in detail, wherein:
Fig. 1 be High Performance Liquid Chromatography/Mass Spectrometry method 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 illustrate of the present invention, but never limit the scope of the present invention.Below with reference to embodiment, further elaborate the present invention, but it will be appreciated by those skilled in the art that the present invention is not limited to the preparation method of these embodiment and use.And those skilled in the art can be equal to replacement, combination, improvement or modify 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 the 85%(v/v of 1.5g) in ethanol-pH4.0 lactic acid buffer solution, obtain drug solution, 0.22 μ m filtering with microporous membrane, add Ovum Gallus domesticus Flavus lecithin E80 7.0g, then add the soybean oil 1.5g after 0.22 μ m filtering with microporous membrane, gnotobasis lower magnetic force stir about 0.5h, dissolve completely to E80, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear completely to bubble, packing, sealing, obtains.
embodiment 2
Get Thymopentin 20mg, be dissolved in 2.0g dehydrated alcohol, obtain drug solution, 0.22 μ m filtering with microporous membrane, add Ovum Gallus domesticus Flavus lecithin E80 6.0g, then add the medium chain length fatty acid triglyceride 2.0g of 0.22 μ m filtering with microporous membrane, the about 0.5h of magnetic agitation, dissolve completely to E80, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear completely to bubble, packing, seal, obtain the high concentration phospholipid slow releasing preparation of Thymopentin.
embodiment 3
Get insulin 100mg, be dissolved in 2.0g 90%(v/v) in ethanol-normal saline solution, obtain drug solution, 0.22 μ m filtering with microporous membrane, add soybean phospholipid 5.5g, then add the soybean oil 2.5g of 0.22 μ m filtering with microporous membrane, the about 0.5h of magnetic agitation, dissolve completely to soybean phospholipid, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear completely to bubble, packing, seal, obtain 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, add Ovum Gallus domesticus Flavus lecithin E80 5.0g, the Oleum Sesami that adds again 3.0g 0.22 μ m filtering with microporous membrane, the about 0.5h of magnetic agitation, dissolve completely to E80, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear substantially completely to bubble, packing, seal, obtain the high concentration phospholipid slow releasing preparation of erythropoietin.
embodiment 5
Get leucine enkephalin 100 μ g, be dissolved in the 85%(v/v of 2.0g) in ethanol-water solution, obtain drug solution, 0.22 μ m filtering with microporous membrane, add hydrogenated soya phosphatide 7.0g, then add the soybean oil of 1.0g 0.22 μ m filtering with microporous membrane, the about 0.5h of magnetic agitation, dissolve completely to hydrogenated soya phosphatide, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear completely to bubble, packing, seal, obtain the high concentration phospholipid slow releasing preparation of leucine enkephalin.
embodiment 6
Get interleukins 16 mg, be dissolved in the 80%(v/v of 1.0g) in ethanol-water solution, obtain drug solution, 0.22 μ m filtering with microporous membrane, add Ovum Gallus domesticus Flavus lecithin E80 7.5g, then add the medium chain length fatty acid triglyceride of 1.5g 0.22 μ m filtering with microporous membrane, the about 0.5h of magnetic agitation, dissolve completely to E80, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear completely to bubble, packing, seal, obtain the high concentration phospholipid slow releasing preparation of interleukin.
embodiment 7
Get interferon 30mg, be dissolved in the 85%(v/v of 1.5g) in ethanol-water solution, obtain drug solution, 0.22 μ m filtering with microporous membrane, add hydrogenated yolk lecithin 6.5g, then add 2.0g through the soybean oil of 0.22 μ m filtering with microporous membrane, the about 0.5h of magnetic agitation, dissolve completely to hydrogenated yolk lecithin, obtain the opaque thick liquid of milk yellow that contains a large amount of bubbles, standingly disappear completely to bubble, packing, seal, obtain the high concentration phospholipid slow releasing preparation of interferon.
embodiment 8
Get leuprorelin acetate 100mg, be dissolved in 1.5g 80%(v/v) in ethanol-water solution, obtain drug solution, 0.22 μ m filtering with microporous membrane, add soybean phospholipid 7.5g, then add 1.0g through the medium chain length fatty acid triglyceride of 0.22 μ m filtering with microporous membrane, the about 0.5h of magnetic agitation, dissolve completely to soybean phospholipid, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear completely to bubble, packing, seal, obtain the high concentration phospholipid slow releasing preparation of leuprorelin acetate.
embodiment 9
Get growth hormone 40mg, be dissolved in the 95%(v/v of 1.5g) in ethanol-water solution, obtain drug solution, 0.22 μ m filtering with microporous membrane, adds dipalmitoyl phosphatidyl choline 6.0g, then adds 2.5g through the ethyl oleate of 0.22 μ m filtering with microporous membrane, the about 0.5h of magnetic agitation, to dissolving completely, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear completely to bubble, obtain clear liquid, packing, seals, and obtains the high concentration phospholipid slow releasing preparation of growth hormone.
embodiment 10
Getting acetic acid Ai Saina peptide 20mg is dissolved in 10ml water for injection, form drug solution, 0.22 μ m filtering with microporous membrane sterilizing, adds injection stage Ovum Gallus domesticus Flavus lecithin E80 3.0g, stirring and evenly mixing under aseptic condition, press the packing of 2ml/ bottle,-40 ℃ freezing 4 hours, on freeze dryer, drain 24 hours, obtain loose lyophilizing phospholipid, sealing, standby.
95% alcoholic solution of 5g and 5g medium chain length fatty acid triglyceride is mixed, and 0.22 μ m filtering with microporous membrane sterilizing, is distributed into 0.4g/ bottle, and sealing is standby.
Face the used time medium chain length fatty acid triglyceride-95% ethanol mixture solution is injected to lyophilizing phospholipid, firmly jolting, dissolves, and obtains.
embodiment 11
Take water soluble drug morphine sulfate 60mg, be dissolved in 2.0g 85%(v/v) in ethanol-water solution, obtain drug solution, 0.22 μ m filtering with microporous membrane, add soybean phospholipid 6.0g, then add 2.0g through the ethyl oleate of 0.22 μ m filtering with microporous membrane, magnetic agitation to soybean phospholipid dissolves completely, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear completely to bubble, obtain clear liquid, packing, sealing, obtains.
embodiment 12
Take fat-soluble medicine nimodipine 40mg, be dissolved in 2.0g dehydrated alcohol, obtain drug solution, 0.22 μ m filtering with microporous membrane, add Ovum Gallus domesticus Flavus lecithin E80 7.0g, then add 1.0g through the medium chain length fatty acid triglyceride of 0.22 μ m filtering with microporous membrane, magnetic agitation is dissolved completely to E80, obtain the milk yellow opaque liquid that contains a large amount of bubbles, standingly disappear substantially completely to bubble, obtain clear liquid, packing, sealing, obtains.
the selection of embodiment 13 concentration of alcohol
Take 4 parts of Ovum Gallus domesticus Flavus lecithins (E80), every part of 7.0g, adds respectively the ethanol of variable concentrations to 10.0g, stirs 2 hours, observes outward appearance, and measures viscosity, the results are shown in Table 1.Visible, concentration of alcohol, more than 80% dissolving well phospholipid, obtains the solution of good fluidity.
The screening of table 1 concentration of alcohol
| Concentration of |
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 |
the impact of the different types of host material of embodiment 14 on preparation viscosity
By table 2 prescription, take respectively different types of phospholipid 7.0g, oily 1.5g and alcoholic solution 1.5g, stir 2 hours, measure viscosity, the results are shown in Table 2.The different types of phospholipid of visible interpolation, oil and alcoholic solution are little to the viscosity influence of preparation.
The impact of the different types of host material of table 2 on 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 consumption screening of embodiment 15 ethanol
The soybean oil and the ethanol solution that by table 3 prescription, take respectively injection Ovum Gallus domesticus Flavus lecithin (E80), filtration sterilization, mix under aseptic condition, is stirred to dissolving, packing, sealing.By below testing 3 methods, carry out local irritation test, tentatively investigate the zest of different dehydrated alcohol consumptions to injection site.
Experimental technique: with experiment 3.Under injection end postscript, inserting needle position and medicine, in subcutaneous position, are observed its variation.The results are shown in Table 3.Visible, in prescription the content of dehydrated alcohol lower than 30% time, preparation to medicine-feeding part do not have zest or zest very little, and reversible.In theory, containing the alcoholic solution replacement dehydrated alcohol of a certain amount of moisture, its toxicity should be lower.
The consumption screening of table 3 ethanol
| Prescription | Phospholipid E80 (g) | Oil (g) | Dehydrated alcohol (g) | Local irritation test result |
| Prescription 1 | 5.0 | 4.0 | 1.0 | Medicine-feeding part is without obvious zest |
| Prescription 2 | 5.0 | 3.0 | 2.0 | Medicine-feeding part is without obvious zest |
| Prescription 3 | 5.0 | 2.5 | 2.5 | Medicine-feeding part is without obvious zest |
| Prescription 4 | 5.0 | 2.0 | 3.0 | Slightly red and swollen, but disappear after 2-3 days |
| Prescription 5 | 5.0 | 1.0 | 4.0 | Medicine-feeding part is red and swollen, and after 5-6 days, skin starts to fester |
[effect experiment]
the viscosity of [testing 1] high concentration phospholipid slow releasing preparation and with the comparison of vesicle type phospholipid gel preparation
Found through experiments, although the concentration of phospholipid is very high in high concentration phospholipid slow releasing preparation, viscosity is within injectable scope, more much lower than the vesicle type phospholipid gel preparation of identical even lower phospholipid concentration.
Vesicle type phospholipid gel preparation prepare reference literature method (M. Brandl, Liposome Technol. 1 (1993) 49 – 65), take respectively Ovum Gallus domesticus Flavus lecithin E80 3.0g, 3.5g, 4.0g, 4.5g, 5.0g, mend and inject water to 10.0g respectively, be stirred to and mix, high pressure homogenize, obtains again.
The viscosity of the vesicle type phospholipid gel preparation of the high concentration phospholipid slow releasing preparation that comparative example 1,2,4,5,8,10 makes and several different phosphate lipid concentrations.Experimental result below:
The comparison of table 4 high concentration phospholipid slow releasing preparation and vesicle type phospholipid gel preparation viscosity
Note: above viscosity number is under 25 ℃ of conditions and records.
From above experimental result, can find out, for high concentration phospholipid slow releasing preparation, its viscosity increases with the rising of content of phospholipid in preparation, but the amplitude changing is less, changes generally little.And for vesicle type phospholipid gel preparation, its viscosity sharply increases with the rising of content of phospholipid in preparation, and its viscosity has reached 11659.4cP when phospholipid concentration reaches 40%, injection difficult, and when phospholipid concentration reaches 50%, almost there is no mobility.And further find, 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, and therefore, high concentration phospholipid slow releasing preparation of the present invention has better syringeability.
the slow release characteristic of [testing 2] high concentration phospholipid slow releasing preparation
Adopt the slow release effect of experiment exam embodiment 1 in animal body.
By embodiment 1(phospholipid concentration, be 70%) product to be expelled to male Wistar rat subcutaneous, regularly get blood, adopt High Performance Liquid Chromatography/Mass Spectrometry method to measure the octreotide acetate content in blood plasma, examine or check its slow releasing function time.
1. the preparation of standard solution
Precision takes 13.18 mg octreotide acetate standard substance, puts in 10 ml volumetric flasks, is dissolved in water, and is settled to scale, is mixed with the storing solution of 1.318 mg/ml.Get this storing solution appropriate, use respectively 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, obtain 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, mix to be placed in 37 ℃ of shaking tables and hatch 30min, after hatching end, add 200 μ l acetonitriles, whirlpool mixes 5min, centrifugal (13500rpm) 5min, supernatant is crossed to 0.22 μ m filtering with microporous membrane, get subsequent filtrate sample introduction.The known drug concentration in blood plasma of take is abscissa, and peak area is vertical coordinate, drawing standard curve, and gained linear regression equation is 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 rat plasma sample is 0.51-10.14ng/ml.
3. the processing of plasma sample
Get 100 μ l plasma samples, first add 25 μ l methanol-waters (1:1 v/v), then add 200 μ l acetonitriles, whirlpool mixes 5min, and centrifugal (13500rpm) 5min crosses 0.22 μ m filtering with microporous membrane by supernatant, gets subsequent filtrate sample introduction.
4. chromatograph and mass spectrum condition
Chromatographic condition chromatographic column is 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,, 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.Cation mode detects, and scan mode is multiple-reaction monitoring (MRM).For the ionic reaction of Mass Spectrometer Method, be respectively m/z 510 → m/z 120 and m/z 510 → m/z 159, wherein m/z 510 → m/z 120 is for quantitative analysis, and 510 → m/z 159 is for qualitative analysis.
5. interior animal experiment
Get 6 male Wistar rats, body weight 200±20g， Sichuan University West China medical experiment animal center provides.Fasting 12 h before administration, press the sample of dosage subcutaneous injection embodiment 1 preparation of 20mg/kg, ad lib and drinking-water after administration.Before administration, after (0h) and administration, 0.5h, 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, centrifugal (4500rpm) 5min, separated plasma, to be measured in-20 ℃ of Refrigerator stores.After above-mentioned plasma sample disposal methods, LC-MS analyzes mensuration.
Measurement result: rat skin lower injection administration can also measure medicine (during medicine, curve is shown in accompanying drawing 2) after 30 days, visible high concentration phospholipid slow releasing preparation has fabulous slow release effect.
the zest research at [testing 3] high concentration phospholipid slow releasing preparation drug administration by injection position
The product of embodiment 1 of take is the zest of representative examination high concentration phospholipid slow releasing preparation to medicine-feeding part.With reference to " chemicals zest, anaphylaxis and hemolytic investigative technique guideline ", carry out local irritation test, and compare with the Comparative formulation product that only contains medicine, phospholipid and ethanol.
The preparation of Comparative formulation: get octreotide acetate 50mg, be dissolved in the 85%(v/v of 3.0g) in ethanol-water solution, obtain drug solution, 0.22 μ m filtering with microporous membrane, add Ovum Gallus domesticus Flavus lecithin E80 7.0g, gnotobasis lower magnetic force stir about 0.5h, dissolves completely to E80, obtains the milk yellow opaque liquid that contains a large amount of bubbles, standingly substantially disappear to bubble, packing, sealing, obtains.
1. test method
Get 8 rabbit, be divided into two groups, each 4, male and female half and half.If normal saline contrast, adopts consubstantiality left and right sides self-contrast method.Test first 24 hours to the administration district of both sides, back lose hair or feathers processing (first with shears by depilation district hair cut to suitable length, then with depilatory, further process).Depilation scope is 3cm * 3cm each side.Before administration, check that skin of unhairing, whether because of depilation damaged, has the skin of damage to test.
Adopt subcutaneous injection administration, adjust inserting needle position, make medicine just in time be expelled to depilation position subcutaneous.The product of 4 rabbit one side injection embodiment 1, opposite side injecting normal saline.The product of another 4 rabbit one side injection comparative examples, opposite side injecting normal saline.Injection finishes under postscript inserting needle position and medicine in subcutaneous position, to observe and carry out medicine-feeding part histopathological examination.After administration, in 72 hours, animal and injection site are carried out to perusal, the observation period finishes latter every group and randomly draws two rabbit and carry out medicine-feeding part histopathological examination.The rabbit staying continues to observe, and carries out histopathological examination, to understand the degree of reversibility of irritative response after 14 days.
2. result of the test
After administration, in 72 hours, animal and injection site are carried out to perusal, just started 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 does not occur other abnormal phenomenas such as erythema or edema.As time goes on, heave gradually and diminish, by 72 hours, do not seen obvious difference.The medicine-feeding part histopathological examination result of 72 hours shows, injects the tissue of high concentration phospholipid slow releasing preparation one side and follows organizing of injecting normal saline one side substantially as broad as long, all there is no obvious medicine-feeding part zest.
Two rabbit that stay after administration during 72h ~ 14 day medicine-feeding part there is not zest phenomenon, and the position of As time goes on losing hair or feathers grows gradually hair and comes.The histopathological examination result of 14 days shows that injection high concentration phospholipid slow releasing preparation is the same with injecting normal saline, does not all occur obvious medicine-feeding part zest.
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, injection site occurs red and swollen, and after 5 days, fester in red and swollen position, shows obvious local toxicity.
Claims (9)
1. an in-situ injection phase change gel slow releasing preparation, it is characterized in that comprising phospholipid, bioactive ingredients, alcoholic solution and oil for injection, gross weight meter based on preparation, wherein content of phospholipid is 50%～(46875/626) %(g/g), bioactive ingredients content is 0.01%～10%(g/g), oil for injection content is 10%～40%(g/g), alcoholic solution content is 30%～5%(g/g), the concentration range of alcoholic solution is for being more than or equal to 80%(v/v) and be less than 100%(v/v); Described phospholipid is selected from natural phospholipid, semi-synthetic phospholipid, synthetic phospholipid one or more combination;
Described oil for injection is selected from 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;
Described alcoholic solution is selected from 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;
Described natural phospholipid is selected from Ovum Gallus domesticus Flavus lecithin, soybean phospholipid or its combination; Described semi-synthetic phospholipid is selected from hydrogenated yolk lecithin, hydrogenated soya phosphatide or its combination; Described synthetic phospholipid is selected from one or more the combination in DPPE, dipalmitoyl phosphatidyl choline, distearoyl phosphatidylcholine, dimyristoyl phosphatidyl choline, DOPE, DPPG, DPPA.
2. an in-situ injection phase change gel slow releasing preparation, it is characterized in that comprising phospholipid, bioactive ingredients, alcoholic solution and oil for injection, gross weight meter based on preparation, wherein content of phospholipid is 50%～(46875/626) %(g/g), bioactive ingredients content is 0.01%～10%(g/g), oil for injection content is 10%～40%(g/g), alcoholic solution content is 30%～5%(g/g); Described phospholipid is selected from natural phospholipid, semi-synthetic phospholipid, synthetic phospholipid one or more combination;
Described oil for injection is selected from 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;
Described alcoholic solution is selected from dehydrated alcohol;
Described natural phospholipid is selected from Ovum Gallus domesticus Flavus lecithin, soybean phospholipid or its combination; Described semi-synthetic phospholipid is selected from hydrogenated yolk lecithin, hydrogenated soya phosphatide or its combination; Described synthetic phospholipid is selected from one or more the combination in DPPE, dipalmitoyl phosphatidyl choline, distearoyl phosphatidylcholine, dimyristoyl phosphatidyl choline, DOPE, DPPG, DPPA.
3. in-situ injection phase change gel slow releasing preparation according to claim 1 and 2, is characterized in that described phospholipid is selected from Ovum Gallus domesticus Flavus lecithin E80.
4. in-situ injection phase change gel slow releasing preparation according to claim 1 and 2, is characterized in that described bioactive ingredients is selected from: insulin, glucagon, alpha-interferon, beta-interferon, interleukin, erythropoietin, promoting sexual gland hormone, tissue-type plasminogen activator, streptokinase, cell growth factor, thrombin, prostaglandin, antithrombase, hirudin, histrelin, growth hormone-releasing peptide-2, His-D-Trp-Ala-Trp-D-Phe-Lys-NH2, cetrorelix, bivalirudin, arginine vasopressin, Schweine-Vasopressin, Desmopressin, leuprorelin, De She Rayleigh, buserelin, triptorelin, goserelin, alarelin, Sermorelin, nafarelin, lutrelin, fertirelin, Hexarelin, octreotide, Exenatide, human glucagon-like-peptide-1, growth hormone, ziconotide, thymosin a1, sincalide, Thymopentin, 1: PN: WO02056903 PAGE: 25 claimed protein, aviptadil, leucine enkephalin, methionine enkephalin,MEK, tetracosactide, gonadotropin releasing hormone, throtropin releasing hormone, growth hormone releasing hormone, Somat, short melanocyte inhibitory hormone, short melanocyte releasing hormone, corticotropin-releasing hormone, secretin, Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2, neurotensin, human brain natriuretic peptide, atrial natriuretic peptide, sleep-inducing peptide, hypermnesia peptide, dynorphin, endorphins, angiotensin, enfuirtide, elcatonin, Thymosin β4, vassopressin, Pitressin Tannate, felypressin, ornipressin, melanotan II, spleen pentapeptide, Pramlintide, vapreotide, thyroliberin, silk relies short cortex 18 peptides, sweet essence is urged cortex 18 peptides, zinc is urged cortex 24 peptides, short cortex 24 peptides, short cortex 25 peptides, short cortex 28 peptides, vasoactive intestinal peptide, Kallidin I, orphanin, asparaginase.
5. in-situ injection phase change gel slow releasing preparation according to claim 1 and 2, is characterized in that described bioactive ingredients is selected from: granulocyte-macrophage colony stimutaing factor, gonadorelin, hypertensinⅠ, angiotensin II, angiotensin II I.
6. in-situ injection phase change gel slow releasing preparation according to claim 1 and 2, is characterized in that described bioactive ingredients is selected from Fondaparinux sodium, morphine, tramadol, oxycodone, fentanyl, lignocaine, dolantin, sorbide nitrate, pentaerithrityl tetranitrate, isosorbide mononitrate, general naphthalene Nore, receive many Nores, indole Nore, Ah 's Nore, Mei Tuonuoer, A Pu Nore, vinegar fourth Nore, nifedipine, verapamil, diltiazem, spectrum Ni Laming, piperazine can former times woods, bepridil, nicorandil, molsidomine, dilazep, trimetazidine, quinidine sulfate, 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, amlodipine, felodipine, nitrendipine, lacidipine, nimodipine, nicardipine, nisoldipine, isradipine, peso Nore, his Nore doubly, prazosin, terazosin, doxazosin, trimazosin, draw 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, melatonin, curcumin, desferrioxamine, idebenone, Tirilazad Mesylate, estradiol, ethinylestradiol, estrone, quinestrol, nilestriol, estradiol valerate, estradiol benzoate, androstenediol, methyltestosterone, testosterone, diethylstilbestrol, diethylstilbestrol propionic ester, diethylstilbestrol phosphate ester, estradiol cypionate, Dai, medroxyprogesterone acetate, megestrol acetate, chlormadinone acetate, gestodene, norethindrone, desogestrel, delalutin, levonorgestrel, norethindrone acetate, norethisterone enanthate, dibasic acid esters etynodiol, Quingestanol Acetate, ground norgesterone, drospirenone, norpregnenes, nomegestrol, trimegestone, diphenhydramine, tripelennamine, chlorphenamine, buclizine, phenindamine, Cyproheptadine, clemastine, azelastine, astemizole, loratadine, cetirizine, mizolastine, teldane, Fexofenadine fourth, Efletirizine, Desloratadine, ebastine, promethazine, pheniramine, brompheniramine, tolpropamine, pyrrobutamine, the pyridine of music score of Chinese operas profit, acrivastine, cimetidine, ranitidine, famotidine, nizatidine, Luo Sha is for fourth, ketotifen, sodium cromoglicate, cyclophosphamide, ifosfamide, carmustine, lomustine, semustine, dacarbazine, procarbazine, temozolomide, chlorambucil, melphalan, cisplatin, carboplatin, oxaliplatin, 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, flutamide, tamoxifen, exemestane, tretinoin, imatinib, gefitinib, erlotinib, Thalidomide, Lenalidomide, magnesium salicylate, choline salicylate, salicylamide, salsalate, diflunisal, indomethacin, sulindac, acetaminophen, aminophenazone, oxyphenbutazone, Phenylbutazone, mefenamic acid, clofenamic acid, diclofenac, ibuprofen, naproxen, fluorine ratio is coughed up sweet smell, fenoprofen, ketoprofen, piroxicam, meloxicam, nimesulide, rofecoxib, celecoxib, colchicine, probenecid, other purine, sulfinpyrazone, benzbromarone.
7. a method of preparing the in-situ injection phase change gel slow releasing preparation described in claim 1 or 2, is characterized in that comprising the following steps:
(1) get bioactive ingredients and be dissolved in appropriate alcoholic solution, then mix with appropriate oil for injection, form drug solution, filtering with microporous membrane sterilizing;
(2) under aseptic condition, get injection stage phospholipid and mix with drug solution, stir phospholipid is dissolved completely, standing a moment, packing, sealed, and obtains to remove the bubble in preparation.
8. a method of preparing the test kit of the in-situ injection phase change gel slow releasing preparation described in claim 1 or 2, is characterized in that comprising the following steps:
(1) get appropriate bioactive ingredients and be dissolved in appropriate pure water or buffer salt solution, form drug solution, filtering with microporous membrane sterilizing, adds injection stage phospholipid, stirring and evenly mixing under aseptic condition;
(2) by above-mentioned semi-solid phospholipid gel packing, lyophilization in the usual way, sealing;
(3) by appropriate alcoholic solution and appropriate oil for injection mixing, filtering with microporous membrane sterilizing, packing, sealing, standby;
(4) product of step (2) and (3), with the mode assembly packaging of test kit.
9. claim 1 or 2 in-situ injection phase change gel slow releasing preparation reduce the application in the in-situ injection phase change gel slow releasing preparation of ethanol side effect in preparation.
Priority Applications (1)
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| CN103705442B (en) * | 2012-09-28 | 2017-12-01 | 上海恒瑞医药有限公司 | Lipid gel pharmaceutical preparation in situ and its production and use |
| CN102866153B (en) * | 2012-09-29 | 2014-08-13 | 郑州安图生物工程股份有限公司 | Prostatitis joint-detection kit |
| JP6134050B2 (en) * | 2013-03-14 | 2017-05-24 | マサチューセッツ インスティテュート オブ テクノロジー | Multilayer injectable self-assembled peptide scaffold hydrogel for long-term sustained release of human antibodies |
| CN104230736B (en) * | 2014-05-26 | 2017-01-04 | 中国人民解放军第二军医大学 | A kind of antiinflammatory, analgesic medicine and preparation thereof |
| CN108771657B (en) * | 2017-06-22 | 2020-11-06 | 四川大学 | Small molecule drug in-situ phase change gel sustained release system and preparation method thereof |
| CN108772042A (en) * | 2018-07-04 | 2018-11-09 | 林思思 | A kind of anthraquinone derivative bonded silica gel stationary phase, preparation method and ginsenoside detection application |
| CN108815514B (en) * | 2018-07-11 | 2022-07-08 | 浙江工业大学 | A hirudin tannate tablet and its preparation method |
| CN111012734B (en) * | 2018-10-09 | 2021-08-20 | 四川大学 | Drug-loaded reticular in-situ phase-change gel sustained-release system and preparation method thereof |
| CN111166890A (en) * | 2018-11-09 | 2020-05-19 | 四川大学 | Slow-release vaccine carrier and preparation method thereof |
| CN109316602A (en) * | 2018-11-13 | 2019-02-12 | 西安力邦医药科技有限责任公司 | The prescription and application of a kind of long-acting analgesic and the compound sustained-released delivery system for promoting wound healing |
| CN111388744B (en) * | 2020-01-15 | 2021-05-18 | 华中科技大学 | Gel storage, preparation method thereof and application of gel storage in preparation of postoperative hemostatic preparation |
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