Background of invention
Pain is a kind of physiology of complexity, psychological activity, it includes two ingredients, first, noxious stimulation acts on bodyCaused pain is felt, when reaction of the individual to the pain of noxious stimulation, also with complicated psychological activity.International pain is groundStudy carefully definition of the meeting (IASP) to pain：Pain is a kind of offending feeling and actual, potential tissue damage, causedEmotional experience；Or to being described made by this damage.Pain belongs to a kind of offending physiology experience, it comes across respectively extensivelyIn the course of disease of kind disease.Pain not only brings discomfort to patient's body, but also also will produce to spirit, psychology, constitution etc.Different degrees of influence directly affects life and the life quality of patient.
Ropivacaine, Bupivacaine are the first-line drug of current clinical local anaesthesia, central nervous system and angiocarpySystem toxicity is relatively low, has been had been more and more widely used in fields such as anesthesia, Postoperative Analgesia Afters, and wherein local anaesthesia effect is summarizedIt is as follows：
One surgery anesthesia
1 intravertebral anesthesia
Epidural ropivacaine is since its broader treatment tolerance interval allows to use higher dosage, to ensureEnhance anaesthetic effect by improving dosage reduce local anaesthetics toxic hazard under the premise of.The discoveries such as Camorcia,The ED50 of 1% Ropivacaine kinesitherapy nerve retardance is 6.1mg (CI 95%：5.1~7.1mg), and when a concentration of 0.1%It is then 9.1mg (CI 95%：7.8~10.3mg), dose increase about 50% when low concentration.Fettes etc. then has found, weighs proportion sievePiperazine cacaine lumbar anesthesia work faster, retardance it is more perfect, retardance after regression time it is shorter.
2 peripheral nerve blocks
The minimum effective concentration of Ropivacaine brachial plexus block is 0.5%, and optimum concentration is then 1%；Increase injection capacity(60ml, 150mg) can shorten the onset time of movement retardance, and Casati etc. thinks that 0.75% Ropivacaine is best suited forSciatic nerve combines retardance with femoral nerve.
Two Pain managements
A large amount of random, perspective studies confirm that Ropivacaine is definite for obstetrical analgesia and labor analgesia, to movementNerve block is light, does not influence stages of labor and newborn, is current Epidural cavity obstetrical analgesia and the ideal drug of inhibiting pain in parturition.SievePiperazine cacaine is treated for URP Postoperative Analgesia Afters, and the Ropivacaine of continuous infusion 0.1% can obtain satisfied analgesic effect, and can subtractThe dosage of few opioid drug.By Ropivacaine Combined With Morphine thoracic epidural controlled analgesia for treating unstable angina pectorisAchieve good effect.
In recent years, in gynaecology's inhibiting pain in parturition, gynecological surgery analgesia, paediatrics analgesia, gerontal patient's analgesia etc., to alleviateIn patient's art, various pain caused by postoperative, cancer and the nervous system disease, clinical application long-acting local anesthetics have useSafety, definite effect, patient restore the features such as fast, therefore clinically bright for the demand growth of long-lasting local anesthesia preparationIt is aobvious, it is worthy to be popularized.
Soybean oil is that a kind of oil extracted is squeezed from soybean, contains a large amount of linoleic acid.Corn oil is by cornEmbryo processing vegetable fat obtained, is mainly made of unsaturated fatty acid.Its Linoleic acid is aliphatic acid needed by human, is structureThe component part of adult somatic cells has important physiological function.Child lacks linoleic acid, and skin becomes drying, and the scales of skin that peel off thickens,Development growth is slow；The elderly lacks linoleic acid, can cause cataract and cardio cerebrovascular affection.Median chain triglyceride oil is from coconut palmIt is extracted in seed oil or from palm oil, it is the mixture of saturated triglycerides, and important is octanoic acids(C8H16O2) and capric acid (C10H20O2).Olive oil is the not heated and change made of the fresh direct cold press of olive fruitHandle.Olive oil is rich in abundant monounsaturated fatty acids-oleic acid, also vitamin A, vitamin B, vitamin D, dimension lifePlain E acetates, vitamin K and polyphenoils etc..
Phosphatide was found from human brain in 1812 by Uauquelin earliest, and Golbley was detached in 1844 from yolkOut, and in 1850 according to Greek Iekithos (yolk) it is named as Lecithin (lecithin).Its main component is phosphorusPhosphatidylcholine (PC), the content of phosphatidyl choline (PC) will purify yolk phospholipid up to 98% or more in the yolk phospholipid of purifyingReferred to as phosphatidyl choline (PC).Using the yolk phospholipid of purifying, phosphatidyl courage in the alprostadil injection listedThe content of alkali is 98% or more.Also there is document that lecithin is claimed to be a kind of mixture, main component is phosphatidyl choline (PC), content80% or so is accounted for, other compositions are lysophosphatidyl choline (L-HPC), phosphatidylinositols (PI), phosphatidylserine (PS), phosphorusAcyl ethanol amine (PE).Phosphatidyl-ethanolamine is also known as cephalin.
Pharmaceutically, lecithin is mainly used as dispersant, emulsifier and stabilizer, is also used as the material of liposome, andIncluded in intramuscular (IM) and intravenous (IV) injection, parenteral absorption preparation and local product.Lecithin is also listed in useFDA in inhalant, IM and IV injections, oral capsule, suspending agent and tablet, rectum, part and vagina preparation is inactiveIn Ingredient Guide.
For weakly basic drugs Ropivacaine, Bupivacaine, can use the content of phosphatidyl choline be up to 95%~100% phosphatide, because it is almost without the acidic phospholipid of contribution negative electrical charge, the blank liquid surface negative charge of preparation is less,When positively charged weakly basic drugs are added, higher positive charge is presented in liquid surface particles, and the stability of liquid can obtainTo guarantee.
Document report, certain active constituents are compound under certain condition with phosphatide, obtain active constituent phosphatide complexes,The more former compound of activity improves significantly, and is effectively improved the absorption of active constituent in vivo, significantly improves its lifeObject validity, changes drug release feature, reduces toxic side effect and dosage.After drug and phosphatide form phosphatide complexes, lipophilicProperty obviously increases, to which bioavilability can be improved.
Oxygen atom in phosphatide on phosphorus atoms in hydroxyl has the tendency that stronger that electronics, nitrogen-atoms have stronger betatopicTendency, therefore, under certain condition, can with the drug of certain structure generate compound.Phosphatide complexes are BombardelliIt was found that a kind of novel drug-loading system, many active constituents of medicine cause difficult absorption, bioavilability low due to physicochemical property itselfThe defects of and limit it and clinically apply；And being formed after compound with phosphatide can then show to live with parent drug difference pharmacologyProperty.The research of drug phosphatide complexes since the 1980s, gradually draws attention, and increasingly extensive depth is applied in researchEnter.Including Phospholipid Complex with Active Constituents from Chinese Materia, metal ion phosphatide complexes, protein and peptide class drug phosphatide complexes withAnd nonsteroidal anti-inflammatory drug phosphatide complexes etc..Wherein non-steroidal anti-inflammatory drugs generally has different degrees of GI irritation to makeWith the compound of formation can be such that the stimulation of these drugs significantly reduces, in addition, nonsteroidal anti-inflammatory drug is ionic mostlyDrug, fat-soluble poor, Transdermal absorption is more difficult, its Determination of oil-water partition coefficient significantly increases after forming compound with phosphatide, toImprove its permeability to skin.
In addition, phosphatide is added in liquid can change the drug release behavior of main ingredient, it can achieve the effect that sustained release.Small point of both sexesSub- substance lecithin can have an impact pectin base colon-site specific drug delivery system, and the addition of lecithin improves pectin gel of zinc ballTo the load medicine Release Performance of Indomethacin and Ketoprofen.This may be that the introducing of lecithin hydrophobic grouping improves the hydrophobic of pectinProperty, the rate of release of drug is reduced, makes the less leakage of drug in dissolution medium.After colon being reached by gel ball, withPectase will accelerate the release of drug to the degradation of pectin in human colon[16], to reach colon locating administrated mesh's.Drug-phospholipid compound can change physical and chemical properties of drugs, with commercial preparation hydroxycamptothecin (HCPT) carboxylic acid salt solution's agentFor reference, the pharmacokinetic trial of phosphatide complexes (HCPT-phytosome) injection of hydroxycamptothecin is carried out, with3P87 programs calculate pharmacokinetic parameter.The two all meets two compartment model, the t of reference preparation1/2, α 4.608min, t1/2, βIt is 128.203 μ gminmL for 30.074min, AUC-1；The t of HCPT phospholipid complex formulations1/2, α 1.208min, t1/2,β is 45.548min, and AUC is 104.337 μ gminmL-1.The pharmacokinetic parameters of the two are fitted with counting Moment Methods,The AUC of HCPT phosphatide complexes0→ t is the 87.1% of reference agent；Elimination rate constant K is 0.016, is the 0.38 of reference agentTimes；The MRT of the two0→ ∞ is respectively 17.064min and 61.081min, and HCPT-phytosome is extended than reference agent3.58 times, the phosphatide complexes of formation make increased retentions of the HCPT in rat body, preparation have extended release kinetics specialSign.
Chinese patent 201380036669 " hydrophobic depot formulations of active constituent and preparation method thereof " and Chinese patentContain castor oil in the slow-released carrier mentioned in 201380036700 " depot formulations of local anesthetic and preparation method thereof ", andCastor oil is low to moderate harm in MSDS reports, has certain irritation and allergy to act on skin and eyes.
Invention content
The present invention provides a kind of analgesic sustained release drug delivery systems, the preferred Ropivacaine of analgesic that the present invention selects orPerson's Bupivacaine, the sustained release drug delivery carrier are phosphatide-soybean oil, and the safety of wherein soybean oil is higher, can avoid castor oilIrritation.Wherein phosphatide is cell membrane constituent, has good biocompatibility, the stabilization of liquid can be effectively ensuredProperty, so that duration of efficacy is significantly extended, dosing interval extends, and administration number of times is less, improves the compliance of patient.
The sustained release drug delivery systems of the present invention, can more extend the release time of drug, and drug release concentration is more steady, partTissue irritation's smaller is greatly reduced the adverse reaction risk of local anesthetic clinical application, highlights good clinical developmentForeground contributes to the compliance of patient, reduces hospital stay and hospital expenses, can be clinically used for the analgesia of open wound section, officePortion's inflammatory pain etc..
For this purpose, the present invention provide it is a kind of with phosphatide-miscible agent-oil for the local anesthetic sustained release preparation of carrier and its preparationMethod.
Local anesthetic sustained release preparation of the present invention, including：As the local anesthetic of active constituent, as sustained releaseThe mixture of phosphatide-miscible agent-oil of delivery system, and optional may also include antioxidant.
Wherein, the mixing of the preferred Bupivacaine of the local anesthetic either one of Ropivacaine free alkali or the twoObject.
Wherein, the phosphatide is selected from natural phospholipid, synthetic phospholipid and hydrogenated phospholipid etc., such as：Egg yolk lecithin, soybeanLecithin, phosphatidyl choline, egg PC, Polyene Phosphatidylcholine, glycerolphosphocholine, hydrogenated soya phosphatide,Hydrogenate yolk phosphide, phosphatidylserine, dioleoyl phosphatidylserine, phosphatidic acid, dipalmitophosphatidic acid, distearoylOne kind in base phosphatidic acid, phosphatidyl-ethanolamine, yolk phosphatidylglycerol, sphingomyelins, phosphatidylinositols, two or moreMixture；It is preferred that egg yolk lecithin, soybean lecithin, phosphatidyl choline, hydrogenated soya phosphatide are a kind of, two or more is mixedClose object；
Wherein, the miscible agent is selected from：One of ethyl alcohol, benzyl alcohol, ethyl acetate, ethyl lactate, the two or the twoAbove with the mixture of Ergol, more preferably include the mixture of ethyl alcohol and Ergol, benzyl alcohol, ethyl alcohol and benzene firstThe mixture of acid benzyl ester；
Wherein, the oil is selected from：Soybean oil, midchain oil, corn oil, olive oil, sunflower oil, tea oil, structure oil, oilAt least one of acetoacetic ester, two or more mixture；
Wherein, the antioxidant is selected from：Vitwas E, cysteine, n-acetyl-L-cysteine, fourthBase hydroxyanisole, butyl hydroxy toluene, propylgallate, tert-butyl hydroquinone, lipoic acid, tea polyphenols, L- Vitamin CsSour palmitate, one kind of glutathione, two kinds or two or more mixtures, further preferred Vitwas E, sulphurSad one or two mixture；
Wherein, in the mixture of phosphatide-miscible agent-oil, content of phospholipid is no more than 50%, Ergol in miscible agentContent be not less than 10%, remaining for oil；
Sustained release preparation of the present invention is prepared by following components in percentage by weight：
Local anesthetic 2-8%；
Sustained release drug delivery systems 91-97%；
Antioxidant 0.1-1%；
Preferably：
Local anesthetic 3-6%；
Sustained release drug delivery systems 93-96%；
Antioxidant 0.1-1%；
Wherein, the weight percent of sustained release drug delivery systems each component is as follows：
Phosphatidase 1 0-50%；
Miscible agent 10-50%；
Remaining is oil 0-80%；
Preferably：
Phosphatidase 1 0-30%；
Miscible agent 15-45%；
Remaining is oil 25-75%；
Sustained release preparation of the present invention, wherein concrete component can be：
1) local anesthetic, ethyl alcohol, Ergol, phosphatide, soybean oil, lipoic acid；
2) local anesthetic, ethyl alcohol, Ergol, phosphatide, midchain oil, Vitwas E；
3) local anesthetic, ethyl alcohol, Ergol, phosphatide, olive oil, lipoic acid；
4) local anesthetic, ethyl alcohol, Ergol, phosphatide, corn oil, Vitwas E；
5) local anesthetic, ethyl alcohol, benzyl alcohol, Ergol, phosphatide, olive oil, lipoic acid；
6) local anesthetic, ethyl alcohol, benzyl alcohol, Ergol, phosphatide, soybean oil, Vitwas E；
7) local anesthetic, ethyl alcohol, benzyl alcohol, Ergol, phosphatide, midchain oil, lipoic acid；
8) local anesthetic, ethyl alcohol, benzyl alcohol, Ergol, phosphatide, corn oil, Vitwas E；
Wherein, phosphatide is preferably one of egg yolk lecithin, soybean lecithin, hydrogenated soybean lecithin, phosphatidyl choline, the two orMixture more than person's the two；
Wherein, phospholipid ratio is no more than 50% (50% refers to that phosphatide accounts for the percent weight in volume g/ml of total preparation)；
Wherein, ethyl alcohol, Ergol, phosphatide, the proportionate relationship of oil are as follows：
1：1：1：2 or 1：3：2：4；
Wherein, ethyl alcohol, benzyl alcohol, Ergol, phosphatide, the proportionate relationship of oil are as follows：1：1：2：2：4；
The present invention further provides the preparation methods of sustained release preparation of the present invention, include the following steps：
It is prepared by A liquids：Precision weighs a certain amount of local anesthetic and is dissolved in the miscible agent of certain volume, ultrasound or stirIt mixes to complete drug dissolution, then adds recipe quantity antioxidant and phosphatide, ultrasound or stirring are completely dissolved to phosphatide, finally addAdd recipe quantity oily, ultrasound or stirs and evenly mixs and prepare required liquid；
B is aseptic subpackaged：Aseptically, the liquid prepared is crossed into film and removes impurity, degerming, packing；The present invention'sMain advantage may be summarized as follows：
1, the solvent in the drug release carrier of phosphatide-miscible agent-oil of the invention selects Ergol, with ethyl alcohol, benzeneMethanol is miscible, and phosphatide-oil can be made to be dissolved into uniform solution, wherein Ergol is indispensable, unique；
2, the phosphatide main component in the drug release carrier of phosphatide-miscible agent-oil of the invention is phosphatidyl choline, and content is highUp to 95%~100%, almost without the acidic phospholipid of contribution negative electrical charge, the positively charged alkalescent medicine Ropivacaine of additionOr Bupivacaine, liquid surface are presented higher positive charge, the stability of liquid have been effectively ensured；
3, the oil of phosphatide-miscible agent-oil in the present invention has selected the higher soybean oil of safety, does not include castor oil,Skin irritation and anaphylaxis are avoided, safety greatly improves；
4, the present invention with phosphatide-miscible agent-oil is slow-released carrier, and duration of efficacy significantly extends, at least 48h withOn, administration number of times is reduced, and is effectively improved the compliance of Clinical practice patient；
5, the phosphatide in the present invention is life basic substance, is the main constituents of cell membrane, has good biologyCompatibility, the addition of phosphatide can reduce the dosage of oil phase, improve local histocompatbility, reduce local anesthetic and faceThe adverse reaction risk of bed application；
6, there is antioxidant in the present invention, can be with stable grease, while having certain suppression to the oxidation impurities of RopivacaineIt makes and uses；
7, the present invention mainly takes the method that cutting part directly uses, so as to avoid clinical patients inconvenience oral drugsAnalgesic is perplexed, while also overcoming oral preparation and working slow disadvantage；
8, invention formulation is oiliness sustained-release preparation, and preparation process is simply controllable, and production cost is greatly reduced；
9, invention formulation is oiliness sustained-release preparation, and pre-filled container is noted for that can be cillin bottle, can also be pre- embeddingEmitter；
10, single-point administration or multi-point injection can be used when invention formulation is administered, single-dose volume is controllable smaller, symbolRequirement of the current oily preparation intramuscular injection to administered volume is closed, irritation, sensitization are smaller, are easy to implement industrialization.
The advantageous effect further illustrated the present invention below by way of experimental data.
The dissolubility test of 1 phosphatide of test example
Precision weighs phosphatide (egg yolk lecithin PC-98T), is separately added into benzyl alcohol, ethyl alcohol, ethyl lactate, Benzyl BenzoateEach 5ml of ester, observation dissolving situation continue to add phosphatide to saturation state, primarily determine phosphatide different molten if dissolving is completeSolubility in agent.As shown in table 1 below.
The solubility test result of 1 phosphatide of table
Experimental result is shown：25 DEG C of room temperature, solubility of the phosphatide in ethyl alcohol, benzyl alcohol is more than 800mg/ml, dissolvingProperty is preferable, takes second place in the solubility of ethyl lactate, Ergol；Therefore, medicine solvent can choose ethyl alcohol, benzyl alcohol,Secondly also optional extracting lactic acid ethyl ester, Ergol, wherein solubility in ethanol is best.
The dissolubility test of 2 oil of test example
Soybean oil is weighed by recipe quantity precision, benzyl alcohol, ethyl alcohol, ethyl lactate, Ergol is separately added into, observes itSituation is dissolved, and by two or three in these types of solvent according to 1:The dissolving situation of oil wherein is investigated in 1 mixing.It is as followsShown in table 2.
The solubility test result of 2 soybean oil of table
| Solvent title | Dissolve situation |
| Benzyl alcohol | Dissolving, solution clarification |
| Ethyl alcohol | Endless fully dissolved, solution are slightly muddy |
| Ethyl lactate | Insoluble, solution is muddy |
| Ergol | Dissolving, solution clarification |
| Benzyl alcohol/ethyl alcohol | Endless fully dissolved, solution are slightly muddy |
| Benzyl alcohol/Ergol | Dissolving, solution clarification |
| Benzyl alcohol/ethyl lactate | Insoluble, solution is muddy |
| Ethyl alcohol/ethyl lactate | Insoluble, solution is muddy |
| Ethyl alcohol/Ergol | Dissolving, solution clarification |
| Benzyl alcohol/ethyl alcohol/Ergol | Dissolving, solution clarification |
Experimental result is shown：25 DEG C of room temperature, solubility of the soybean oil in benzyl alcohol, Ergol is best, in ethyl alcoholIn dissolubility take second place；It is miscible in benzyl alcohol/Ergol, benzyl alcohol/ethyl alcohol/Ergol, ethyl alcohol/ErgolSolubility in agent is preferable.In conjunction with the conclusion in test example 1, phosphatide can be compared with good dissolving in alcohol solvent.Therefore it deduces mostExcellent miscible agent is ethyl alcohol/Ergol, benzyl alcohol/ethyl alcohol/Ergol, but need to further be tested by following experimentCard.
The intersolubility of 3 blank medicine-carried system of test example is tested
Using ethyl alcohol, benzyl alcohol, Ergol as medicine solvent, be added after phosphatide dissolving respectively with soybean oil, oliveOil, corn oil, midchain oil progress are miscible, add stabilizer, observe mutually immiscible situation.
The blank medicine-carried system of experiment includes following：
Ethyl alcohol/benzyl alcohol/soybean oil/egg yolk lecithin/Vitwas E
Ethyl alcohol/benzyl alcohol/olive oil/soybean lecithin/Vitwas E
Ethyl alcohol/benzyl alcohol/corn oil/phosphatidyl choline/Vitwas E
Ethyl alcohol/benzyl alcohol/midchain oil/hydrogenated soybean lecithin/Vitwas E
Benzyl alcohol/Ergol/soybean oil/egg yolk lecithin/Vitwas E
Benzyl alcohol/Ergol/olive oil/soybean lecithin/Vitwas E
Benzyl alcohol/Ergol/corn oil/phosphatidyl choline/Vitwas E
Benzyl alcohol/Ergol/midchain oil/hydrogenated soybean lecithin/Vitwas E
Ethyl alcohol/Ergol/soybean oil/egg yolk lecithin/Vitwas E
Ethyl alcohol/Ergol/olive oil/soybean lecithin/Vitwas E
Ethyl alcohol/Ergol/corn oil/phosphatidyl choline/Vitwas E
Ethyl alcohol/Ergol/midchain oil/hydrogenated soybean lecithin/Vitwas E
Benzyl alcohol/ethyl alcohol/Ergol/soybean oil/egg yolk lecithin/Vitwas E
Benzyl alcohol/ethyl alcohol/Ergol/olive oil/soybean lecithin/Vitwas E
Benzyl alcohol/ethyl alcohol/Ergol/corn oil/phosphatidyl choline/Vitwas E
Benzyl alcohol/ethyl alcohol/Ergol/midchain oil/hydrogenated soybean lecithin/Vitwas E
Blank medicine-carried system with preferable intersolubility is：
Ethyl alcohol/Ergol/soybean oil/egg yolk lecithin/Vitwas E
Ethyl alcohol/Ergol/olive oil/soybean lecithin/Vitwas E
Ethyl alcohol/Ergol/corn oil/phosphatidyl choline/Vitwas E
Ethyl alcohol/Ergol/midchain oil/hydrogenated soybean lecithin/Vitwas E
Benzyl alcohol/ethyl alcohol/Ergol/soybean oil/egg yolk lecithin/Vitwas E
Benzyl alcohol/ethyl alcohol/Ergol/olive oil/soybean lecithin/Vitwas E
Benzyl alcohol/ethyl alcohol/Ergol/corn oil/phosphatidyl choline/Vitwas E
Benzyl alcohol/ethyl alcohol/Ergol/midchain oil/hydrogenated soybean lecithin/Vitwas E
Experimental result prompts：The intersolubility having had containing Ergol in blank medicine-carried system, wherein ethyl alcohol/benzene firstAlcohol/Ergol, ethyl alcohol/Ergol the blank system of miscible agent mix and can preferably dissolve each other with soybean oil and phosphatide,It is consistent with conclusion (of pressure testing) before.
4 Ropivacaine free alkali of test example, dissolubility test of the Bupivacaine free alkali in medicine-carried system
Precision weighs Ropivacaine free alkali respectively, and Bupivacaine free alkali is appropriate, is separately added into ethyl alcohol, benzyl alcohol, benzeneEach 5ml such as benzyl formate, midchain oil, soybean oil, corn oil, olive oil, observation dissolving situation continue to add if dissolving is completeDosing object primarily determines solubility range of the drug in different solvents, as shown in table 3 to saturation state.
3 Ropivacaine free alkali of table, dissolubility test of the Bupivacaine free alkali in medicine-carried system
| Solvent title | Ropivacaine free alkali (mg/ml) | Bupivacaine free alkali (mg/ml) |
| Ethyl alcohol | 162 | 134 |
| Benzyl alcohol | 226 | 210 |
| Ergol | 72 | 88 |
| Midchain oil | 10.8 | 9.6 |
| Soybean oil | 9.7 | 10.8 |
| Corn oil | 10 | 11.2 |
| Olive oil | 9.5 | 9.9 |
Experimental result is shown：25 DEG C of room temperature, solubility of the Ropivacaine free alkali in ethyl alcohol, benzyl alcohol are more than160mg/ml, dissolubility is preferable, takes second place in the solubility of Ergol；Solvent can choose ethyl alcohol, benzyl alcohol, Benzyl BenzoateThree kinds of ester.Bupivacaine free alkali ethyl alcohol, benzyl alcohol solubility more than 120mg/ml, dissolubility is preferable, in benzoic acidThe solubility of benzyl ester is taken second place.
5 pharmaceutical preparation stability test of test example
On the basis of test example 1,4 experience of test example 2, test example 3 and test example, in addition other components in preparation, 2It places 48 hours for~8 DEG C and observes its solubility property.The results are shown in Table 4.
4 pharmaceutical preparation stability test result of table
Experimental result is shown：Ropivacaine and Bupivacaine are in preferred drug release carrier system-ethyl alcohol/methanol/benzoic acidBenzyl ester, ethyl alcohol/Ergol miscible agent mixed with soybean oil, olive oil, any oil phase of midchain oil relatively stablize, 2~8DEG C place 48h after without drug be precipitated, have preferable dissolubility.
The Ropivacaine sustained release preparation sensory nerve blockade test (hot plate method) of the different prescriptions of test example 6
The selection of animal
By hot plate method to 230~250g of weight, healthy female Wister rats carry out testing preceding screening.By hot plate literTemperature is to (56 ± 1) DEG C, and a wherein metapedes for rat is placed on it, and in addition a metapedes is then in laboratory room temperature conditionIn, observe and record its tested time retracted enough.Then in kind measure an other metapedes for rat.Every metapedesIt alternately measures, is spaced 5 minutes, each measure is averaged three times, is denoted as Basic Pain Threshold value.As paw withdrawal time measurement result is more than5.0 seconds, then the rat Basic Pain Threshold value is undesirable, should give rejecting.
Grouping and administration
54 rats are taken to be tested, this experiment is divided into 9 groups, and each group is made a living respectively manages brine group, Ropivacaine injectionLiquid group, -1 group of Ropivacaine sustained release preparation, -2 group of Ropivacaine sustained release preparation, -3 group of Ropivacaine sustained release preparation, Ropivacaine- 4 group of sustained release preparation, -5 group of Ropivacaine sustained release preparation, -6 group of Ropivacaine sustained release preparation, -7 group of Ropivacaine sustained release preparation(specific prescription is shown in Table 5 with specification), each group injects relative medicine around Rat Right sciatic nerve-trunk respectively, and each group dosage is equalFor 10mg/kg.
The different Ropivacaine sustained release preparation prescriptions of table 5
Method and evaluation
Local anaesthetics has apparent retardation for rat sciatic nerve, feels that retardance becomes after investigating the administration of each group ratChange situation, and carries out comprehensive compare and evaluation.To avoid the occurrence of experimental animal body injury, as rat is more than in hot-plateThere is not paw withdrawal phenomenon yet in 15s, should be denoted as 15s away from hot plate, and by experimental result.
Statistical method
For statistical analysis with SPSS10.0 softwares, t is examined between chi-square criterion and group, and P < 0.05 are statistically significant, P< 0.01 indicates that tool is statistically significant.
The evaluation of result of sensory nerve retardance
Experimental result such as table 6, in the local anaesthesia experiment investigation of Ropivacaine sustained release preparation, sensory nerve retardance is shownApparent slow release effect.The Ropivacaine sustained release preparation of different prescriptions has certain duration analgesic activity, and certain formulations canContinue 48 hours even 72 hours or more, Ropivacaine sustained release preparation group has significance difference compared with Ropivacaine injection groupIt is different.In clinical analgesia therapy, need to mitigate pain by the feeling retardation of local anaesthetics in some cases, while not wishing againHoping influences the proper motion function of patient, can be by meeting such treatment requirement to the further research of this slow release product.
The different prescription Ropivacaine sustained release preparation rat intramuscular injection kinesitherapy nerve residence time statistics (n=6) of table 6
Influence of the different prescription Ropivacaine sustained release preparations of test example 7 to pig wound healing
Experiment packet and dosage：Ba-Ma mini pig, 12-25kg, male, 54 or so, adaptability is raised 2-3 daysAfterwards, it is screened according to weight and is divided into 9 groups, every group is 6, and each group is respectively control group, Ropivacaine injection group, Ropivacaine- 1 group of preparation, -2 group of Ropivacaine preparation, -3 group of Ropivacaine preparation, -4 group of Ropivacaine preparation, -5 group of Ropivacaine preparation,- 6 group of Ropivacaine preparation, -7 group of Ropivacaine preparation, dosage are 0.5ml/.
Basic experiment solution processes：For experimental animal after carrying out screening and being grouped according to weight, each experimental group bar horse is small-sizedPig first carries out back depilation, and next day operation establishes 2cm*1cm Ba-Ma mini pigs back full-thickness defects wound model and is calculated as D0, clapsAccording to record.Random grouping, by group administration, administering mode is wound multiple spot intramuscular injection nearby.
Observation index：D1, D3, D7, D14, D21 different time points observe each group Ba-Ma mini pig feelings upon administration respectivelyCondition, and score with recovery situation according to wound healing area.
The different prescription Ropivacaine preparations of table 7 score (n=6) to Ba-Ma mini pig wound healing situation
| Grouping | D1 | D3 | D7 | D14 | D21 |
| Control group | - | 18.36% | 50.24% | 85.65% | 97.58% |
| Ropivacaine injection group | - | 20.15% | 54.21% | 84.53% | 97.64% |
| - 1 group of Ropivacaine sustained release preparation | - | 19.65% | 51.68% | 83.67% | 98.55% |
| - 2 group of Ropivacaine sustained release preparation | - | 19.52% | 53.23% | 82.94% | 99.01% |
| - 3 group of Ropivacaine sustained release preparation | - | 22.35% | 52.10% | 83.28% | 97.34% |
| - 4 group of Ropivacaine sustained release preparation | - | 20.89% | 50.13% | 83.67% | 96.95% |
| - 5 group of Ropivacaine sustained release preparation | - | 19.35% | 49.85% | 86.95% | 97.38% |
| - 6 group of Ropivacaine sustained release preparation | - | 18.95% | 48.57% | 87.56% | 98.65% |
| - 7 group of Ropivacaine sustained release preparation | - | 18.06% | 48.95% | 88.65% | 99.32% |
Evaluation of result：Comparative evaluation is carried out for different groups, different observation point Ba-Ma mini pig wound healing situations,The result shows that different prescription Ropivacaine sustained release preparation prescriptions have no significant effect Ba-Ma mini pig wound healing.
8 muscle irritation of test example is tested
Whether normal first 1 week of experiment observes state of mind, appetite, body temperature of rabbit etc..It is noted using rabbit quadriceps muscle of thighMethod takes healthy 9 rabbit to be divided into 3 groups, and every group 3, male and female have concurrently, 2~2.5kg of weight, respectively on the left of the 1st group of rabbitThe injection 1mL of -6 group of Ropivacaine sustained release preparation is injected at hind leg quadriceps muscle of thigh, at the 2nd group of rabbit left rear limb quadriceps muscle of thighThe injection 1mL of -7 group of Ropivacaine sustained release preparation is injected, sterile physiological salt is injected at the 3rd group of rabbit left rear limb quadriceps muscle of thighWater 1mL.2,7 and 15d takes each 1 execution of every group of rabbit respectively upon administration, and solution takes quadriceps muscle of thigh, longitudinally slit, observation notePenetrate the tissue change of position muscle, such as congested, red and swollen, denaturation, necrosis.Lesion diameter is measured, is stimulated according to musculature anti-The evaluation criteria answered is evaluated.In addition it takes both sides injection site musculature, after conventional treatment, makees histopathologic examination.Result evaluation standard is shown in Table 8.When the average response score value of rabbit provides that the product can be used as at 2 grades hereinafter, can determine that meetIntramuscular injection, average response score value are more than 2 grades, can determine that be against regulation, which is not available as intramuscular injection.
The reaction classification of 8 musculature of table
| The order of reaction | Stimulate the reaction |
| 0 grade (-) | The musculature of injection site is compared with compareing position musculature without any difference |
| 1 grade (+) | The muscle groups of injection site are woven with hyperemia, and diameter is in 0.5cm or less |
| 2 grades (++) | The musculature of injection site is red and swollen, congested, and diameter is in 1cm or so |
| 3 grades (+++) | The musculature of injection site is red and swollen, congested, blue, gloss disappears, it is seen that necrosis region |
| 4 grades (++++) | The musculature of injection site is red and swollen, congested, blue, gloss disappears, and necrotic extent diameter reaches 0.5cm |
| 5 grades (+++ ++) | Every reaction of the musculature of injection site is more serious, there is large stretch of necrosis |
Test result
Method is noted using rabbit quadriceps muscle of thigh, upon administration 2,7 and 15d, eye is seen without swelling, touches injection site without scleroma.Dissect deferred shares of stock musculus quadriceps, it is longitudinally slit, the response situation of injection site musculature is observed, determines the order of reaction (being shown in Table 8),It is averagely classified as 1 grade, is less than 2 grades, saline control group is 0 grade, and sterile saline, muscle are injected to rabbit quadriceps muscle of thighTissue is identical as normal muscle tissues；Injection site does not occur after -6 2,7 and 15d of group injection of injection Ropivacaine sustained release preparationThe irritative responses such as musculature redness, hyperemia, the performance of the rabbit state of mind are good.Inject -7 group of Ropivacaine sustained release preparationThere is the irritative responses such as musculature redness, hyperemia, the performance of the rabbit state of mind in injection site after 2,7 and 15d of injectionIt is not very good.Microscopically observation muscle fiber band is clear, structural integrity, no denaturation, necrosis and inflammatory reaction, with lifeReason saline control group relatively has no apparent pathological change.Note can be effectively reduced after phosphatide is added in Ropivacaine slow-release injectionIrritation when penetrating.
9 injection of table is to rabbit quadriceps muscle of thigh test result