WO1992007551A1

WO1992007551A1 - Process for producing fat emulsion

Info

Publication number: WO1992007551A1
Application number: PCT/JP1991/001510
Authority: WO
Inventors: Junzo Seki; Hirofumi Yamamoto; Shuji Yamane; Yutaka Takahashi; Kouichi Ushimaru
Original assignee: Nippon Shinyaku Co., Ltd.
Priority date: 1990-11-06
Filing date: 1991-11-05
Publication date: 1992-05-14

Abstract

A process for producing a fat emulsion wherein the emulsion particle has a mean diameter of 10 to 100 nm, characterized by shortening the emulsification time through the addition of sodium chloride as an emulsification aid in an amount of 0.01 to 0.2 % (w/v) based on the whole emulsion in an arbitrary stage of the production process. This process also serves to reduce the decomposition of the emulsion components as a result of shortening the emulsification time.

Description

明

Production of fats and milks

Technical field

The present invention relates to an improved technique for producing a fat-and-oil emulsion having an average particle size of 10 nm to 100 nm.

Background technology

The fat-resistant emulsion having an average particle size of 10 to 10 OOnro has an effect of improving the transferability of a drug from blood or an application site to a diseased tissue, and has excellent characteristics.

A fat emulsion having such an average particle size has the property of avoiding uptake (RBS) by the reticuloendothelial system. In addition, compared to a fat emulsion having a diameter of lOOmn or more, it can maintain a high concentration in the blood, and easily leak out of the blood vessel from a site where vascular permeability is enhanced. It is said that blood vessels have various pore systems (pore system, small pore systems up to 9 nm in diameter and large pore systems with diameters of 25 to 70 nro). It is known that the permeability is further increased.) There is a site called), and there are other intercellular spaces, and in various lesions such as inflammation, pain, atheromas, etc., vascular permeability is enhanced. At such sites, more ultrafine emulsion particles than the blood vessels selectively leak through the large volume system and migrate into the diseased tissue. At the same time, the drug contained in the particles also moves into the lesion. As a result, the drug can be easily and selectively transferred to the lesion, so that the drug concentration at the lesion can be further increased, and the effect can be increased. In normal cells, the presence of the small pore system described above allows the permeability of particles smaller than lOnra. Prevents the transfer of such small particles from blood vessels to normal cells. From the above, ΙΟππ! Fat emulsions having an average particle size of about 70 nm to about 70 nm, and fat emulsions having an average particle size of about 10 to 100 nm in consideration of the pore size tolerance of the poi system and the particle size distribution of the fat emulsion particles. It is the best for improving drug transportability (see JP-A-2-203).

Average particle size is ΙΟηπ! The present inventors have already found that it is important to limit the quantitative ratio of the simple lipids and the phospholipids, which are the constituents, in order to produce a fat emulsion having a 〜ππι. It contains 0.5 to 30% (w / v) of simple lipids, 0.15 to 2 times the weight of simple lipids (by weight), and an appropriate amount of water. This is achieved by:

Another important requirement in producing a fat emulsion having an average particle S of 10 nm to LOOnm was the production process.

In order to produce such a fat emulsion, ordinary machines, tools, equipment, and the like can be used. For example, all components, including drugs, can be prepared using a manton-gaurine-type 髙 -pressure homogenizer, Although it could be manufactured by a method of forming finely by a fluidizer, an ultrasonic homogenizer, or the like, the use of these required a sufficient pressure and sufficient filling time.

For example, when using an ultrasonic homogenizer (Branson model 185), it takes 60 minutes or more for 10 ml of the coarse emulsion, and when using a microfluidic distiller, it takes 10 minutes or more. () 101 to require even more than 60 minutes during operation with a E Ryo圧5 ^ / cm ^8.

Since many of the lipids constituting the fat emulsion according to the present invention contain unsaturated fatty acids, they are denatured by oxidation, and May be toxic. Applying heat or applying physical pressure, such as mixing, may help to alter such lipids over a long period of time.

In the above manufacturing process, a process of applying a physical pressure such as heat or mixing has to be continued for a relatively long time so far, and there is a strong possibility that the lipid may be altered.

In addition, large-scale facilities were required, the risk of foreign matter being mixed was high, and there were economic disadvantages.

Disclosure of the invention

The present inventors have made various studies for the purpose of solving the above technical disadvantages, and as a result, have reached the present invention.

The gist of the present invention is that the average particle diameter is Ι Οηη! In the step of producing a fat emulsion having a drug size of ~ 100 nm and containing a drug, 0.01% to 0.2% (w / v) of sodium chloride in the whole fat emulsion is emulsified. It is used as an agent.

It is common practice to use isotonic agents in the process of producing fat emulsions, but substances that dissolve in water can cause agglomeration of the particles due to the tight distribution of the emulsion particles. Therefore, for the purpose of maintaining the average particle size, it is not preferable to use an electrolyte material such as sodium chloride as the isotonic material, for example, glycerin, mannitol. It was common practice to use non-analogues such as ト. Therefore, when trying to achieve the object of the present invention, it has been conventionally recognized that the use of sodium chloride as an emulsifying aid is inappropriate.

However, by accident, when sodium chloride was present in the process of producing the fat and oil emulsion of the present invention, it was surprising that The present inventors have found that a phenomenon completely opposite to the common sense of the above occurs, and have completed the present invention.

By adding the sodium chloride of the present invention before the emulsification step, the time required for the emulsification step can be reduced to 1/2 to L / 3. This can reduce the total processing energy required for emulsification. This means that if the emulsification step is continued within the same time, the processing energy can be reduced as compared to before. Further, from the above, (1) the deterioration of the fat emulsion in the processing step is prevented beforehand, and (2) the present invention can prevent the abrasion of equipment for emulsification and reduce the maintenance cost. To obtain the effect of this.

In the present invention, only sodium chloride I) can be selectively used. In the present invention, other clays, eg, calcium chloride, magnesium chloride, sodium carbonate, etc. cannot be applied.

The amount of sodium chloride used in the present invention is suitably 0.01% to 0.2% (w / v) of the total amount of fat to be emulsified.

The effect cannot be obtained at a horn degree of less than 0.01%, and the effect becomes opposite when added at more than 0.2%, and the emulsion grains formed are not stable. Therefore, the specific effects of the present invention can be obtained only in the range of 0.0% to 0.2%. Preferably, it is 0.01 to 0.1%.

• In the present invention, sodium chloride can be added in any step of emulsion production. It is possible to mix it with lipids before emulsification, or it may be dissolved in ice before emulsification. Alternatively, it may be added during the emulsification step.

In the present invention, sodium chloride is added before the end of the emulsification step. There is a need. Before the end of the emulsification step, it can be added to simple lipids or to phospholipids.

In the above, in addition to adding sodium chloride itself, a substance containing chloride and sodium salt is added to neutralize sodium chloride during the process of producing a fat emulsion. An operation that produces the same result as the addition of sodium chloride as a result of the formation of the aluminum also constitutes the present invention.

Examples of the lipid used in the present invention include simple lipids derived from natural plant minerals, derived lipids, complex lipids, and mixtures thereof. For example, any of pure lipid, conductive lipid, or complex lipid derived from egg yolk, soybean, cotton, or the like, or any of pure synthetically produced simple lipid, conductive lipid, or complex lipid may be used.

Examples of the simple lipid include those which can be usually used such as refined soybean oil, cottonseed oil, rapeseed oil, sesame oil and the like.

Examples of phospholipids include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, and the like. These hydrogenated products can also be used. A particularly preferred representative example is egg yolk purified lecithin.

The drug to which the present invention can be applied is not particularly limited as long as it is pharmaceutically acceptable. Even insoluble or poorly soluble drugs can be used. In the present invention, the drug easily forms an emulsion.

Drugs to which the emulsion of the present invention can be applied include, for example, anti-inflammatory drugs, analgesics, anti-allergic drugs, antibiotics, chemotherapeutic drugs, anti-cancer drugs, Anti-viral agent, Anti-atherosclerotic agent, Anti-lipemic agent, Anti- * 瘙 agent, Immunomodulator, Pectin, Radical remover, Bronchodilator, Hypnotic, Tranquilizer, Local anesthetic, Fat-soluble vitamins, new medicines and the like can be mentioned. Examples of these include, for example, ancitabine, fluorouracil, mitomycin (:, mitomycin C, furnesylate, mitomycin C, funnesyl acid amide, nonyloxycarbonylmytomycin (:, Mystomycin C derivatives such as cholesteryl oxyglysilmytomysin C, etc., limofur, phtofuryl palmitate, 5—fluorosilyl myristate, ryolimycin, daunomycin. , Anti-cancer drugs such as cytarabine derivatives such as aclarubicin hydrochloride, macra j revisin, vinblastine, pinklistin, cytarabine B-fatty acid ester, mittan, estramustine, and dichloroflavan Viruses, steroids, e.g. dexamethas Palmitic acid ester, nodocortisone palmitic acid ester, blednisolone palmitic acid ester, dexamethasone stearic acid ester, methylprednisolone, no, lame.tazone, fluocinolone Methazone Probionate, Hide Cortisone Fatty Acid Ester, Ridoson, Spi-D-Nolactone, and Non-Speroids, such as Ibuprofen, Flufenamic Acid, Ketoprofen, Phenacetin, Rintivirin, Rinominopirin, Phenylbutazone indole acetic acid ester, Bif: t-Tyrylpropionic acid derivative, Indomethacin, Indomethacin Ethoxycarbonyl methyl ester , India Metal Sintering Lilyester, Kim Cholingo Cetyl esters, dichloroethylene Roff Enaku, and the like Ryo cetyl salicylic acid and its derivatives. Antiallergic agents such as tranilast, ketotifen, and zelastine can also be used. Examples of antibiotics and chemotherapeutic agents include tetracycline base, erythromycin, midecamysin, ammho τ-lysine Bs, minocycline, miconazole and the like. Can be mentioned. PGE! And PGA! Are examples of brostaglandin agents. PGA 1 alkyl ester, PGE i alkyl ester, PGE j derivative, PGI ₂ derivative, PGD derivative and the like can be used. Difamine X-hydramine, olphinadrine, chlorphenoxamin, chlorulf: L-histamine, anti-histamine such as L-lamin, promethazine, meclizine, cipro-buta-zine, and mouth-xazidine. Agents can also be mentioned. In addition, local anesthetics such as lidocaine, penzocaine, dantrolene, cocaine, tetraccaine, virokine, mebira-kin, and derivatives thereof can also be mentioned. . Hepatopathy ameliorating agents, such as mallotilate, glycyrrhetinic acid, acetyltillyl retinoic acid ethyl ester, glycyrrhetinic acid methyl ester, and anti-inflammatory agents, such as farnesol, geraniol, gefarnet, tepreno , Brown towels, and softcon. Central nervous system drugs, such as penobarbital, methaqualone, haine, diazepam, medazeham, frazeham *, clotiazepam, etizolam, mecridin, buclidin, ajifu Methine, methamphetamine, imimiramin, chlorimiramin, imitriptyline, mianserin, trimetadione, phensuximid, tetrabe Enzamide, penzunamide, camphor, dimorpholine, striki, chlorepromazine, 'promethazine, brochlorazine, mequitazine, triflupromazine, levomepromazine, ziff JL Nido And their derivatives. Examples of bronchodilators include vestophilin and other theophylline derivatives, methyl efdrine and the like. Anti-cholinergic agents, such as benztrobin, physostigmine, at-mouth bottle, scopolamine, etc., parasympathetic β-blockers, such as oxyphencycline, virenzebine, etmidrin, etc. Calcium blockers, for example, diltiazem, diphezibine, perapamil, etc., opener, for example, dipenzamine, phenoxybenzamine, etc., antitussives, for example, nosibi bin, dextromethorphan , Pentoxibelin, pembu α-perin, etc., therapeutic agents for prostate hypertrophy, eg, gastron, oxenden, etc. , Spartin, pavulin, etc., antihyperlipidemic drugs such as crofibrate, simfibrate, blobcol, etc. It can be also mentioned. In addition, for example, vaccines such as polyamino acids, vitamins, dilazeb hydrochloride, ubitecarenone, flavoxate, cyclosporin II, influenza, dibenzthion, diphenirabiline, phenova Renewal, metadion, tofisobam, limonene, etc. can also be mentioned. Fat-soluble vitamins include vitamin ミ and its derivatives ビ, vitamin E and its derivatives, vitamin K and its derivatives, vitamin D and their derivatives, and the like. .

In addition, guaiazulene and essential oily crude drugs, such as, for example, oil of quinnin, oil of primrose, time oil, turpentine oil, eucalyptus oil, palm oil, poppy oil, camellia oil and the like can be mentioned.

New drugs include, for example, compounds labeled with radioisotopes, and radiopharmaceuticals, iodized poppy oil fatty acids, which are X-ray contrast agents. Steal and the like can be mentioned.

The drug to which the present invention can be applied is not particularly limited, as described above. However, when judging from the characteristics of the emulsion properties derived from the particle size of the emulsion, inflammation, blood, vascular Or drugs that are involved in the immune system are generally desirable.

The drug concentration in the fat emulsion of the present invention can be appropriately reduced according to the biological activity of the drug. In addition, the concentrations of the emulsion constituents and the drug in the preparation using the present invention can be appropriately determined as desired. is there

The shape and particle size of the fat emulsion of the present invention can be easily confirmed by communication with an electron microscope, a light scattering type particle size analyzer, a membrane filter, or the like.

Optional components of the fat emulsion of the present invention include additives and auxiliary substances used in general injections. For example, antioxidants, preservatives, stabilizers, tonicity agents, buffers and the like can be mentioned. The required and optimal amounts of these additives, trapping substances, etc. can be varied according to the purpose.

The fat emulsion of the present invention can be sterilized (eg, sterilized by filtration or high-pressure steam) as necessary, and enclosed in an amble together with nitrogen gas. It can be freeze-dried if necessary. The lyophilized material can be reconstituted by adding an appropriate solution.

The emulsion of the present invention can be provided as it is, but if necessary, it is easy to remove sodium chloride added as an emulsifying aid by a general de-wetting operation.

The fat emulsion of the present invention is generally administered intravenously, As in the case of conventional products, it can also be administered as an injection intraarterially, intramuscularly, intracavitary, subcutaneously, or the like. The emulsion of the present invention can also be formulated and used as eye drops, drops *, inhalants, orally administered drugs, bladder injections, external preparations or suppositories. In this case also, additives such as pharmaceutically acceptable bases and excipients can be cited as optional components.

The best way to carry out the invention

The present invention will be described in more detail with reference to Test Examples and Examples of the present invention under £ 1. Test example 1

30 mg of dexamethasone palmitate, 0.6 g of refined soybean oil and 0.5 g of refined egg yolk lecithin were mixed and dissolved in 100 ml of a mixed solution of black mouth form Z methanol (1/1, v / v). Thereafter, the solvent was completely removed under reduced pressure using a rotary evaporator. To this, add Permanent Solution 8 containing 5% glucose and 0.05% sodium chloride, and stir with a homogenizer to obtain a coarse emulsion. Then, an aqueous solution containing 5% of glucose and 0.03% of sodium chloride was added, and the volume was adjusted to 10 m to obtain a sample. Similarly, a control sample was prepared by roughly dispersing a 5% aqueous solution of dalcose containing no sodium chloride and then dispersing the solution. Each of them was emulsified under the same conditions using an ultrasonic homogenizer (Branson model 185) under ice cooling, and the temporary particle size was measured. The results are shown in Table 1.

In the test sample, the particle size already reached a plateau in 40 to 60 minutes, but this phenomenon did not occur in the control sample even in 60 minutes. In the sample Already had a particle size of 43 nm in 30 minutes, but the control sample showed a value of 43 nro only after a lapse of 60 minutes, twice as long. Table 1 Comparison with the sample without sodium chloride

Test example 2

Amphotericin B (3 mg), purified soybean oil (0.5 g) and purified egg yolk lecithin (0.4 g) ^ dimyristylphosphatidylglycerol (0.1 g) mixed with a mouth-form / methanol (1/1, v / v) mixture 100 m After mixing and dissolving in £, the solvent is completely removed under reduced pressure using a rotary evaporator. to this

Then, 8 m of a 0.1% aqueous sodium chloride solution was added, and the mixture was stirred with a homogenizer to obtain a coarse emulsion, which was used as a sample. Similarly, a 0.9% sodium chloride aqueous solution (physiological saline) was used as a crude dispersion to serve as a control sample. Each of them was emulsified under the same conditions using a microfluidizer (end pressure: 5 kgZcrf) under ice-cooling, and the particle size over time was measured. The results are shown in Table 2.

The size reduction of the sample reached a plateau in 40 to 60 minutes, but this phenomenon did not occur in the control sample even in 60 minutes. In addition, the particle size of the sample sample already reached 62nra in 20 minutes, whereas the control sample had a particle size of 60 Table 20.9 Comparison with the sample that yielded sodium chloride

Example 1

3 g of dexamethasone balmitate, 50 g of refined soybean oil and 20 g of refined egg yolk lecithin are heated and mixed at about 60, and then maltose is added to 10% and sodium chloride is added as emulsifying aid to 0. Add 500% of 1% ice solution and mix with a homomixer to obtain a coarse emulsion. The crude emulsion was cured under high pressure with a Manton-Gaurin homogenizer to obtain an emulsion having emulsion particles having a particle size of 10-100 LOOmii. This was freeze-dried according to a conventional method. Example 2

After mixing and dissolving 30 mg of difugibin, 0.6 g of refined soybean oil and 0.5 g of refined egg yolk lecithin in lOOmg of α-form form Z methanol (l / l, v / v) mixture, rotary -Completely remove the solvent under reduced pressure with a vaporizer. 0 Add 8 m of an aqueous solution containing 5% glucose and 0.05% sodium chloride, and stir with a homogenizer to obtain a coarse emulsion. And a permanent solution containing 5% dalcose and 0.05% sodium chloride. 〗

13

After adding to a volume of 10 mL, the mixture was emulsified with an ultrasonic homogenizer (Branson model 185) for 30 minutes under ice-cooling to obtain an emulsion having 10 to 10 nm of emulsion particles. This was freeze-dried according to a conventional method.

Example 3

30 mg of chomhotericin B, 5 g of refined soybean oil and 5 g of refined egg yolk lecithin are homogenized in a mortar, and 10 g of maltose is added to the mixture. To this, add 80m of injection solution, and stir with a polon homogenizer to obtain a coarse emulsion. To this, 10 mg of sodium chloride was added as an emulsifying aid, and after dissolution, water for injection was added, and the volume was adjusted to 100 mL.The emulsion was emulsified with a microfluidizer under permanent cooling. An emulsion with a diameter of 10 to 100 nm was obtained. This was freeze-dried according to a conventional method. The condition of the dried cake was very good, and no defects such as chipping, cracking, and convergence were observed. When water for injection was added and redissolved, the dissolution was completed very quickly, and the emulsion particles were completely restored with no change in the particle size of the emulsion particles after the dissolution.

Example 4

2 g of miconazole, 20 g of refined soybean oil and 30 g of refined egg yolk lecithin are heated and mixed at about 60, and an aqueous solution containing 20% sucrose and 0.07% sodium chloride is added thereto. In addition, the volume is adjusted to 100 ml, and then stirred with a homomixer to obtain a coarse emulsion. Then, the coarse emulsion was emulsified under reduced pressure with a Micro n-Dizer to obtain an emulsion having an emulsion particle size of 10 to 10 O O rm. This was freeze-dried according to a conventional method.

Example 5

1 mg of cyclosporin A, 0.5 g of cholesterol luterate and 0.5 g of purified egg yolk lecithin mixed with black-mouthed form Z methanol (1/1, v / v) 100 After mixing and dissolving in the solvent, completely remove the solvent under reduced pressure with a single evaporator. To this, add 8 ml of a permanent solution containing 5% trehalose and 0.03% sodium chloride, and stir with a homogenizer to obtain a crude emulsified solution. Then, a permanent solution containing 5% of trehalose and 0.03% of sodium chloride was added, and the volume was adjusted to 10 ml. Then, the mixture was treated with an ultrasonic homogenizer (Branson model 185) for 60 minutes. The emulsion was emulsified to obtain an emulsion having a particle diameter of 10 to 100 nm. This was freeze-dried according to a conventional method.

Example 6

Amphotericin B 3 nig, 0.5 g of refined soybean oil, 0.4 g of refined egg yolk lecithin, and 0.1 g of dimyristyl phosphatidylglycerol were added to form mouth Z methanol (1/1). , _y / _v ) After mixing and dissolving in a mixed solution of 100 ml, the solvent is completely removed under reduced pressure using a rotary evaporator. To this is added 8 mL of a 0.1% sodium chloride ice solution, and the mixture is stirred with a homogenizer to obtain a coarse emulsion. The emulsion was emulsified with an ultrasonic homogenizer (Branson model 185) for 60 minutes to obtain an emulsion having an emulsion particle size S of 10 to 10 OOnra. After 1 g of maltose was added to this and dissolved, water was added to adjust the volume to 10πι £. This was freeze-dried according to a conventional method. The condition of the dried cake was extremely good, and no chipping, cracking, shrinkage, etc. were observed. When water for injection was added and redissolved, the dissolution was completed very quickly, and the emulsion particles after the dissolution were completely restored without any change in the particle diameter.

Claims

551 15 Scope of request

(1) G.5 to 30% (w / v) of simple lipids in total, 0.15 to 2 times (weight ratio) of phospholipids than pure fat, and an appropriate amount of ice are required. In producing a fat emulsion with an average particle size of 10 nm to 100 rini, which is characterized by having a constituent component of 0.1% to 0.2% (w / v) of sodium chloride, emulsified sodium chloride A method for producing a fat emulsion, which is used as an auxiliary.