FR2552679A1 - VESICULAR COMPOSITIONS BASED ON QUATERNARY AMMONIUM SALT AND SURFACTANT AGENT - Google Patents

Abstract

VESICULAR COMPOSITIONS CONSISTING OF THE FOLLOWING CONSTITUENTS A AND B: A A QUATERNARY AMMONIUM SALT OF THE TWO-CHAIN TYPE, OF GENERAL FORMULA: (CF DRAWING IN BOPI) IN WHICH R AND R ARE EACH ONE HYDROCARBON RADICAL HAVING FROM 10 TO 24 ATOMES CARBON, R AND R ARE EACH A HYDROCARBON OR HYDROXYHYDROCARBON RADICAL HAVING 1 TO 4 CARBON ATOMS, OR A BENZYL RADICAL, AND X IS AN ANION SERVING AS A COMPLEMENTARY ION TO THE QUATERNARY AMMONIUM ION; B ONE OR MORE SURFACE-ACTIVE AGENTS CHOSEN FROM THE GROUP INCLUDING COMPONENTS HAVING FORMULAS OF THE TYPE: (CF DRAWING IN BOPI) THE WEIGHT RATIO OF CONSTITUENT A TO CONSTITUENT B IS FROM 100: 1 TO 100: 100. THE B CONSTITUENTS ARE USED TO STABILIZE VESICLES, AND THE COMPOSITIONS, ACCORDING TO THE INVENTION, MAY FORM VESICLES STABLE OVER A LONG PERIOD.

Description

This invention relates to vesicular compositions, and more

  particularly vesicular compositions which are capable of forming stable vesicles on a

long period.

  It is well known that phospholipids, in particular lecithin, which are important constituents of the "biomembrane", form in the water hollow double-layered vesicles called liposomes. This liposome is a hollow sphere with a double layer of lipids. It can contain various chemical substances in its hollow part, and thus has a close resemblance to the red blood cells with respect to the structure. In this sense, the liposome has been studied as a model of red blood cell or as a cellular model and

  plays an important role in the study of the biomembrane.

  In recent years, attention has focused on the liposome as a carrier of drugs in vivo. More particularly, the liposome, which is capable of containing various chemicals in its hollow part, may be considered as a kind It has been reported that when a drug is dosed after inclusion in the liposome, the in vivo metabolism of the drug is suppressed and the drug is retained in the living body for a long time, maintaining its effectiveness. Therapeutic (see, for example, FEBS Letters, Vol 36, No. 3, page 292, 1973), that the side effects of the drug, allergy for example, are removed (see, for example, FEBS Letters, Vol 45, 71, 1974), and that the distribution of the drug in the various organs changes (see, for example,

  Eur J Biochem Vol 47, page 179, 1974).

  As indicated above, the liposome exhibits good properties as a carrier of drugs in vivo, and in particular its property of modifying the distribution of the drug in the organs gives the possibility of selectively acting a drug on the affected oranes, for example, most anti-cancer drugs cause side effects because they affect not only cancer cells but also healthy normal cells If it is possible to selectively activate an anticancer drug on cancer cells by administering it after incorporating it into the liposome, this mode of administration is considered very useful In practice, a few attempts have shown good results (see, for example, Collection of Summaries of Lectures to the Meeting of the Japanese Cancer Society,

page 8, (1976)).

  On the other hand, a suitable conductive material, such as an alkylamine, is incorporated in the double-layered portion at the same time as a photoreductive substance, such as cupric ions, is placed in the internal liquid.

  liposome and a photooxidative substance, such as ascorbic acid, in the liposome outer liquid.

  Then, by irradiating this liposome with light, it becomes possible to concentrate a specific type of ions of the external liquid in the internal liquid, depending on the property of the conductive material which is incorporated in the double layer membrane. is possible to collect, as resources, specific types of elemental ions

from seawater.

  As indicated above, the liposome gives rise to a novel and time-bound technique in the administration of drugs, for example. However, since phospholipids which are capable of forming the liposome are substances derived from living bodies, there exists large limits to their chemical structures, so there are

  disadvantages such as the difficulty of conferring them various functions and relatively poor chemical stability.

  In order to eliminate such liposome limits from phospholipids, studies have recently been made to form liposome-type vesicles from synthetic surfactants. It has been confirmed to date that several types of surfactants have the property of These vesicles of surfactants have the hollow double-layered vesicular structure similar to that of the liposome and may therefore be applicable as

  carriers of drugs as previously indicated.

  However, the known surfactant vesicles are all prepared by dispersing the surfactants in water and irradiating the dispersion, with ultrasonic waves for example. As the resulting vesicle structure is unstable, the vesicles can not be stored over a long period of time. More particularly, in the case where the dialkyl phosphates used in the present invention are used to obtain vesicles according to the known method described above, the vesicle structure will disintegrate in a few days, the vesicle solution becoming cloudy or

gelling.

  For the use of surfactant vesicles as carriers of drugs or for other applications, it would be essential to improve stability, so there is a strong demand for the development of techniques.

to improve it.

  In view of the foregoing circumstances, the present inventors have conducted intensive studies to obtain stable vesicles over a long period of time and, as a result of these studies, they have found that vesicles obtained from a a specific type of quaternary ammonium salt having the property of forming vesicles, a type of surfactant, and water, were stable over a long period of time.

  invention was made on the basis of this discovery.

  Accordingly, it is an object of this invention to provide a vesicular composition comprising the following two components (A) and (B), the weight ratio of the components (A) and (B) being in the range of 100: 1

at 100: 100,

  (A) a quaternary ammonium salt of the two-chain type, of general formula (I)

R 1 R 3

  Wherein R 1 and R 2 are hydrocarbon radicals having 10 to 24 carbon atoms, R 3 and R 4 are hydrocarbon or hydroxyhydrocarbon radicals having 1 to 4 carbon atoms, or benzyl radicals, and Xe is an anion serving as an ion complementary to the quaternary ammonium ion, (B) one or more surface-active agents chosen from the group comprising the compounds of formulas (a) to (q), LR 6 R 4 l N Xe (a)

/ \

  R 3 R 5 j R 60 (AO) n H (b) -R 6 -O (AO) n H (c) HO (AO) p (C 3 H 60) q (AO) r H Hn (OA)

) O (AO) LH

H O (AO) m H (d) (e) (f)

CH 20 COR 6

l

HOH 2 C-C-CH 2 OH

I

CH 2 OH

R 6 COO (AO) R 10 l

CH 2 OR 6

CHO Rll CH 2 OR 12

CH 200 CR 7

I

CHOR 8 CH 20 R 9

  (g) (h) (i) CH 2 O (AO) 1 COR 6 CHO (AO) m H CH (AO) n H (AO) m HR 6 N (AO) n H (j) (k) AOH /

R 6 CONA (1)

  AOH R 16 R 15 -N O (m) I R 17 R 3 R 13 C-N -N-R 14 (n) II t

O R 4

  R 16 R 15 -N (A) tc OO (OO 5 N 1 A 1 <o) IIR 17 R 15 NH (A) t COOM (p) R 16 R 15 -N - (A) t SO 3 e (q) R 17 in which, R 5 is a hydrocarbon or hydroxyhydrocarbon radical having 1 to 4 carbon atoms, or a benzyl radical, R 6 is a hydrocarbon radical having 8 to 36 carbon atoms, R 7 is a hydrocarbon radical having 5 to 23 carbon atoms, R 8, R 9 and R 10 are hydrogen atoms or acyl radicals having from 6 to 24 carbon atoms (provided that when R 7 and R 8 are both in the same molecule, at least one of them being a hydrogen atom), R 11 and R 12 are hydrogen atoms or hydrocarbon radicals having from 6 to 24 carbon atoms (provided that at least one of the two is a hydrogen atom), R 13 is a hydrocarbon radical having 9 to 23 carbon atoms, R 14 is a hydrocarbon or hydroxyhydrocarbon radical having 1 to 24 carbon atoms, or a benzyl radical, R 15, R 1 And R 17 are hydrocarbon radicals having 1 to 24 carbon atoms, A is an alkylene radical having 2 to 4 carbon atoms, M is an alkali metal ion, a monoethanolammonium ion, a diethanolammonium ion or an ion. triethanolammonium, 1, m and N are O or are integers from 1 to 150, p, q and r are integers from 1 to 150, t is an integer of 1 to 4, and

  Xe has the same definition as above.

  It was already known that quaternary ammonium salts of the two-chain type, which constitute ingredient (A), form vesicles, as described in Japanese Patent Application Publication No. 53-134784 and as reported in J Am Chem Soc, Vol 99, p. 3860 (1977), J. Colloid Sci Interface, Vol 65, No. 1, page 155 (1978) and J. Am Chem Soc, Vol 101, p. 4030 (1979) However, it was not known not, until it was discovered for the first time by the authors of the present invention, that the stability of these vesicles could be maintained over a long period of time.

  period by incorporating component (B) into component (A).

  Examples of component (A) of the present invention include didecyldimethylammonium salt, didodecyldimethylammonium salt, ditetradecyl dimethylammonium salt, dihexadecyldimethylammonium salt,

  dioctadecyldimethylammonium salt, the di-icosyl salt

  dimethylammonium salt, didecyl-N-hydroxyethyl-N-methylammonium salt, dodecyl-N-hydroxyethyl-N-methylammonium salt, diterradecyl-N-hydroxyethyl-N-methylammonium salt, dihexadecyl-N-hydroxyethyl-N-methylammonium salt, dioctadecyl-N-hydroxyethyl-N-methylammonium salt, di-eicosyl-N-hydroxyethyl-N-methylammonium salt, and similar salts As regards the complementary ions of component (A), they are not critical but it is best if it is

  halogen ions, methylsulfate ions or ethylsulfate ions.

  The surfactants that are used as

  Components (B) in the practice of the invention are considered to have a vesicle stabilizing action.

  The surfactant of formula (a) is a cationic surfactant, the surfactants of formulas (b) to (m) are nonionic surfactants, and the surfactants of formulas (No. ) in (q) are amphoteric surfactants In these constituents (B), the alkylene oxide added to the nonionic surfactants of

  Formulas (b) to (m) is preferably ethylene oxide.

  In the vesicular compositions according to the invention, the weight ratio of mixture of the constituents (A) and (B) is important and must be in the range of 100: 1 to 100: 100 Outside this range, or else it does not form

  no vesicles, or the vesicles formed are unstable.

  To prepare vesicular compositions according to the invention, the constituents (A) and (B) are dissolved in solvents capable of dissolving the two constituents, then the mixture is stirred to obtain a homogeneous solution and the

  solvent according to any known procedures.

  In order to obtain vesicles from the composition thus obtained according to the invention, it is sufficient to suspend the vesicular composition in water and

  subject this suspension to ultrasonic irradiation.

  Apart from this procedure, vesicular solutions can be obtained, for example, by a procedure of dissolving the vesicular composition in a water-miscible solvent such as ethanol, and strongly injecting the solution into water. or by a procedure of solubilizing the vesicular composition with water-soluble surfactants and dialyzing it to remove the surfactant. The vesicular solution prepared from the vesicular composition according to the invention should preferably have a concentration of from 1 to 50% by weight (hereinafter simply referred to as%), or better still from 5 to 30% when this concentration exceeds 50%, the viscosity of the solution becomes too great, which can be a source of difficulty in the preparation and use of the vesicles. In the case where the concentration is less than 1%, there are no difficulties of preparation and of use, but the costs of transporting and conditioning the vesicular solution

  increase, which makes it uneconomic.

  The most reliable method currently known in the art for confirming the presence of vesicles in the thus prepared vesicle solution is to observe it under an electron microscope using a negative staining technique. This negative staining technique is a method of increasing the density. electrodes of the hydrophilic portions of surfactants capable of forming vesicles, by means of phosphotungstic acid or uranyl acetate, and to blacken these hydrophilic parts in the practice of the invention, the electron microscopic vesicle formation Vesicular solutions containing vesicles are transparent and very fluid On the other hand, the solutions in which the compounds used do not form vesicles but give a multi-layered structure are gelled and opaque with a very poor fluidity As a result , 35 in the stability test vesicles (which will be described later in detail), it is possible to judge the stability of a vesicular composition by preparing a vesicular solution by ultrasonic irradiation, and by observing the transparency and the fluidity of the solution as a function of time. A simple method for confirming vesicle formation is nuclear magnetic resonance spectroscopy (NMR). The relaxation time and the width of the absorption line in H-or C 13-NMR of the vesicles are in fact clearly different for compositions vesiculars and non-vesicular compositions. More particularly, with the vesicles, the relaxation time is prolonged and the absorption line is narrow and clean. In the case of non-vesicular compositions, the relaxation time is short and the absorption line However, the presence of the double-layer structure that characterizes the vesicles can not be directly confirmed by NMR In order to confirm for sure the presence of vesicles, it is necessary to

  make an observation under the electron microscope.

  The present invention will be illustrated further in

  examples of which should not be considered as limiting the present invention.

Example 1

  Vesicular compositions were prepared from quaternary ammonium salts of the two-chain type, and various surfactants listed in Table 1, for

  check the formation and stability of the resulting vesicles.

  Preparation of vesicular compositions 10 g of a quaternary ammonium salt of the two-chain type, dissolved in chloroform, and 1 g of each surfactant were mixed and stirred until a homogeneous solution was obtained. removed chloroform with an evaporator to obtain a powder of the vesicular composition. Formation and Confirmation of Vesicle Presence 10 g of the previously obtained powder composition were mixed with 90 g of water and stirred to obtain a viscous, opaque, gel-like composition. This composition was subjected to 60 C and ultrasonic irradiation at 100 W and 25 k Hz for approximately one hour. The resulting solution was observed under an electron microscope, which confirmed the formation of vesicles. the vesicles had formed had a good transparency and a good fluidity. Vesicle Stability Test The vesicle solutions thus obtained were stored for 3 months in a C-thermostatic chamber. Their stability was evaluated by comparing the condition of each solution after 3 months with the condition it had. Immediately after preparation, with regard to transparency and fluidity The standards for evaluating the formation and stability of vesicles are as follows: A: the condition after preservation is exactly the

  same as the state immediately after preparation, the vesicular structure retaining perfectly.

  B: compared to the solution immediately after the preparation, there is only a slight increase in viscosity, the vesicular structure being conserved so

almost perfect.

  C: the solution becomes more viscous and takes a considerable opacity, the vesicular structure hardly surviving. D: the solution immediately after the preparation

  becomes completely gelled and opaque, no vesicular structure being noted.

  :: Despite the ultrasonic irradiation, it does not form

no vesicles.

Table 1

  Results of Observation Agent Agent Agent Surfactant Selmonio Active Active Ammonium Acyl Activated Acensio: 2-Quaternary N 2 N 3 Type Two-chain N 02 N 3 Didecyldimethyl 10 Ammonium Chloride BBB ABB Didodecyldimethylammonium Chloride Diterradecyl dimethyl ammonium chloride ABA Dihexadecyl dimethylammonium chloride ABA Dioctadecyl dimethyl ammonium chloride ABA Di-icosyldimethylammonium chloride BBA Didecyl-N-hydroxy-20-ethyl-N-methylammonium chloride BBB Didodecyl-N-hydroxyethyl-N-methylammonium chloride BBB Diterradecyl-N-hydroxyethyl-N-methylammonium chloride AAA Dihexadecyl chlorideN25 hydroxyethyl-N-methylammonium AAA Dioctadecyl chlorideNhydroxyethyl-N-methylammonium AAA Di-icosyl-N-hydroxyethyl-N-methylammonium chloride ABA Didecyl-N-benzyl chloride N-methylammonium BBB Didodecyl-N-benzyl-N-methylammonium chloride BBB Ditertradecyl chloride Nbenzyl-N-methylammonium BAA Dihexadecyl-N35 benzyl-N-methylammonium chloride BAA Dioctadecyl-N-benzyl-N-methylammonium chloride BAA / - 13 - Table 1 (continued) Dicyosyl-N-benzyl chloride BBA N-methylammonium Dioleyldimethylammonium chloride ABA Dioleyl chloride -N-hydroxymethyl-N-methylammonium ABA Dioleyl-N-benzyl-N-methylammonium chloride ABA :: Surfactants Surfactant nl (CH 2 CH 2 0) m HC 18 H 37 N (CH 2 CH 2 O) n H (N, N-dipolyoxyethylene-N-stearylamine, m + N = 20) Surfactant # 2

C 18 H 37 '1

CH 3 N 3 / N

CH 31 01 C

  c C He CH 3 (trimethylstearylammonium chloride) surfactant n 3 CH 3

C 17 H 35-C-N -N -H 37 C 18

He I

O CH 3

  Comparative Example 1 10 g of each of the quaternary ammonium salts of the two-chain type, which are shown in Table 1, were dissolved in 90 g of water and then vesicles were formed according to the procedure of US Pat. Example 1 The vesicle solutions were stored in a thermostatic chamber at 20 ° C.

  for 3 months to check their condition after storage.

  In each case using one of the 21 quaternary ammonium salts, it was found that vesicles had formed,

  but after the 3-month storage period, the solutions became gelled and opaque by losing their vesicular structure.

Example 2

  Dioctadecyldimethylammonium chloride and the surfactants listed in Table 2 were used to prepare the vesicle compositions according to the procedure of Example 1. The stability of the vesicles obtained from these compositions was determined.

  The results are given in Table 2.

  Table 2 Surfactant 2 n Trimethylstearylammonium chloride Stearyl alcohol Polyoxyethylene stearyl ether (n = 100) Nonylphenyl ether polyoxyethyl ether (n = 10) Block copolymer of polyoxyethylene and polyoxypropylene (p = 10 iq = 10, r = 10) Stearate of polyoxyethylene sorbitan (l + m + n = 5) Sorbitan stearate Pentaerythritol stearate Polyoxyethylene stearate (n = 7) Stearyl glyceryl ether Stearic acid monoglyceride Polyoxyethylene glycerine stearate (l + m + n = 8) Polyoxyethylene stearyl amine (m + n = 20) Stearic acid diethanolamide Stearyl dimethylamine distearyl aminimide Stearyl dimethylcarbobetaine (t = 1) Sodium dodecylaminocarboxylate (t = 1) Stearyl dimethyl sulfobetaine (t = 3) Observation results

B A A B

A B B A A A A A A A B A B B B

_ _

Example 3

  The twelve quaternary ammonium salts of the two-chain type which are shown in Table 3 were used, and the three surfactants used in Example 1, in combination, to prepare 36 vesicular compositions according to the procedure of US Pat. Example 1 in order to

  check the stability of the vesicles obtained from the respective compositions.

  The results are given in Table 3.

Table 3

  Results of Observation Se Agent Agent Agent Surfactosurfactant Ammonium Ammonia Active: Active Two-Chain Active Element nln 2 n 3 Dioctadecyl Dimethylammonium Bromide ABA Dioctadecyl Dimethylammonium Methyl Sulfate BAA Dioctadecyl Dimethylammonium Ethyl Sulfate BBB Dioctadecyl Dimethylammonium Acetate BBB Dioctadecyl-N-hydroxyethyl-N-methylammonium bromide ABA Dioctadecyl-N-hydroxyethyl-N-methylammonium methyl bromide BBA Dioctadecyl-NB AA hydroxyethyl-N-methylammonium ethyl alcohol BAA Dioctadecyl-N-hydroxyethyl-N-methylammonium acetate BBB Dioctadecyl-N-benzyl N-methylammonium bromide BBA Dioctadecyl methylsulphate N35 benzyl-N-methylammonium BBA Dioctadecyl-Nbenzyl-N-methylammonium Ethylsulfate BBA Dioctadecyl-N-benzyl-N-methylammonium acetate BBB

  __________________________________________________________B_

Example 4

  Dioctadecyl dimethyl ammonium chloride and the surfactants used in Example 1 were mixed in different proportions in Table 4, and a total of 39 vesicular compositions were prepared according to the procedure of Example 1. used these compositions to determine whether vesicles were formed and whether these vesicles were stable.

are given in Table 4.

Table 4

  Results of Observation Agent Agent Agent Agent surfactant surfactant surfactant ratio active active ingredient (by weight) n / l n 2 N 03 Dioctadecyl dimethyl chloride / ammonium agent / surfactant

/ 0.5 C C C

/ 1.0 B B B

/ 1,1 B B B

/ 1.25 B B B

100 / 1.67 B B A

/ 2.0 A B A

/ 2.5 A B A

/ 5.0 A B A

/ 10 A B A

100/20 A B A

/ 50 A B B

/ 100 B B B

/ 200 C

Claims (2)

  A vesicular composition comprising the following two components (A) and (B), the ratio of component (A) to component (B) being from 100: 1 to 100: 100, (A) a quaternary ammonium salt of the type with two chains of the general formula (I) 10 R 1 R 3 N X (I)   R 2 R 4 in which R 1 and R 2 are hydrocarbon radicals having 10 to 24 carbon atoms, R 3 and R 4 are hydrocarbon or hydroxyhydrocarbon radicals having 1 to 4 carbon atoms, or radicals benzyl, and Xv is a quaternary ammonium ion-complementary anion, (B) one or more surfactants selected from the group consisting of the compounds of formulas (a) to (q), RN \ JX (a) R 3 R   R 60 (AO) n H (b) R 6 X> O (AO) n H (c) HO (AO) p (C 3 H 60) q (AO) r H (d) / ICH 200 CR 6   H (OA) OO (AO) 1H (e) H O (AO) m H 2552679 CH 20 COR 6 J HOH 2 C-C-C H 120 H (f) CH 2 OH R 6 COO (AO) n R 10 (g) CH 20 R 6 I CHO R 11 (h) CH 2 OR 12 CH 2 OOCR 7 CHOR 8 (i) CH 20 R 9 CH 20 (AO) 1 COR 6   CHO (AO) m H (j) CH 2 O (AO) n H (AO) m H R 6 N (k) (AO) n H HAE R 6 CONA (1) HAE R 16   R 15-1 5 o (m) R 5 -N j 0) R 17 R 3 R 13 C-Ne-Ne-R 1 4 Il 1 0 R 4 R 16 R 15 -N - (A) t COO 8 (o) R 17   And wherein R 5 is a hydrocarbon or hydroxyhydrocarbon radical having 1 to 4 carbon atoms, or a radical benzyl, R 6 is a hydrocarbon radical having 8 to 36 carbon atoms, R 7 is a hydrocarbon radical having 5 to 23 carbon atoms, R 8, R 9 and R 10 are hydrogen atoms or acyl radicals having 6 to 24 carbon atoms (provided that when R 7 and R are both in the same molecule, at least one of them is a hydrogen atom), R 11 and R 12 are hydrogen or hydrocarbon radicals having 6 to 24 carbon atoms (provided that at least one of them is a hydrogen atom), R 13 is a hydrocarbon radical having 9 to 23 carbon atoms, R 14 is a hydrocarbon or hydroxyhydrocarbon radical having 1 to 24 carbon atoms, or a benzyl radical, R 15, R 16 and R 17 are hydrocarbon radicals having 1 to 24 carbon atoms, A is an alkkykene radical having 2 to 4 carbon atoms, M is an alkali metal ion, a monoethanolammonium ion, a diethanolammonium ion or a triethanolammonium ion, 1, m and N are equal to O or are integers from 1 to 150, p, q and r are integers from 1 to 150, t is an integer from 1 to 4, and   X has the same definition as above.   Composition according to Claim 1, characterized in that the complementary ion X of each of the constituents (A) and (B) is a halogen ion, a methylsulphate ion or a ethylsulfate ion.