A sustained-release composition containing a hydroxynaphthoic acid salt of a biologically active substance and a biodegradable polymer, a method of its production, and a pharmaceutical composition containing said sustained-release composition.

The present invention relates to a sustained-release composition of a biologically active substance, and a production method thereof.
Japanese Patent Unexamined Publication No. 97334/1995 discloses a sustained-release preparation comprising a biologically active peptide or salt thereof and a biodegradable polymer having a free carboxyl group at one end and a production method thereof.
The patent publications for GB2209937, GB2234169, GB2234896, GB2257909 and EP626170A2 disclose compositions based on a biodegradable polymer containing a separately prepared water-insoluble salt, such as a pamoate of a peptide or protein, or production methods therefor.
The patent publication for WO95/15767 discloses the embonate (pamoate) of cetrorelix (LH-RH antagonist) and a production method therefor, and describes that the peptide-releasing profile of this pamoate remains the same as in its use alone, even when included in a biodegradable polymer.
To provide a novel composition that contains a high concentration of biologically active substance, and that is capable of a controlled rate of release.
After extensive investigation aiming at resolving the above problem, the present inventors found that when a biologically active substance is incorporated in high concentration in a composition by allowing the biologically active substance and the hydroxynaphthoic acid to be co-present during formation of the composition, and when both are included in a biodegradable polymer, the biologically active substance is released at rates differing from those of the biologically active substance from the counterpart composition of the biologically active substance and hydroxynaphthoic acid prepared in the absence of the biodegradable polymer, which rate of release is controllable by choosing the appropriate kind of biodegradable polymer. The inventors conducted further investigations based on this finding, and developed the present invention.
Accordingly, the present invention provides:
(1) a sustained-release composition containing a biologically active substance or salt thereof, a hydroxynaphthoic acid or salt thereof, and a biodegradable polymer or salt thereof,
(2) a sustained-release composition according to term (1) above wherein the biologically active substance is a biologically active peptide,
(3) a sustained-release composition according to term (2) above wherein the biologically active peptide is an LH-RH derivative,
(4) a sustained-release composition according to term (1) above wherein the hydroxynaphthoic acid is 3-hydroxy-2-naphthoic acid,
(5) a sustained-release composition according to term (1) above wherein the biodegradable polymer is an xcex1-hydroxycarboxylic acid polymer,
(6) a sustained-release composition according to term (5) above wherein the xcex1-hydroxycarboxylic acid polymer is a lactic acid-glycolic acid polymer,
(7) a sustained-release composition according to term (6) above wherein the content ratio of lactic acid and glycolic acid is 100/0 to 40/60 mol %;
(8) a sustained-release composition according to term (7) above wherein the content ratio of lactic acid and glycolic acid is 100/0 mol %,
(9) a sustained-release composition according to term (6) above wherein the weight-average molecular weight of the polymer is about 3,000 to about 100,000,
(10) a sustained-release composition according to term (9) above wherein the weight-average molecular weight of the polymer is about 20,000 to about 50,000,
(11) a sustained-release composition according to term (3) above, wherein the LH-RH derivative is a peptide represented by the formula:
5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z (SEQ ID NO.:1)
wherein Y represents DLeu, DAla, DTrp, DSer(tBu), D2Nal or DHis(ImBzl); Z represents NH-C2H5 or Gly-NH2,
(12) a sustained-release composition according to term (6) above, wherein the terminal carboxyl group content of the polymer is 50-90 micromol per unit mass (gram) of the polymer,
(13) a sustained-release composition according to term (3) above, wherein the molar ratio of the hydroxynaphthoic acid or salt thereof and the LH-RH derivative or salt thereof is from 3 to 4 to 4 to 3,
(14) a sustained-release composition according to term (13) above, wherein the LH-RH derivative or salt thereof is contained at 14% (w/w) to 24% (w/w),
(15) a sustained-release composition according to term (1) above, wherein the bioactive substance or salt thereof is very slightly-soluble in water or soluble in water,
(16) a sustained-release composition according to term (1) above, which is intended for injection,
(17) a method of producing the sustained-release composition according to term (1) above, comprising removing the solvent from a mixture of a bioactive substance or salt thereof, a biodegradable polymer or salt thereof, and hydroxynaphthoic acid or a salt thereof,
(18) a method of producing the sustained-release composition according to term (17) above, comprising mixing and dispersing a bioactive substance or salt thereof in an organic solvent solution containing a biodegradable polymer or salt thereof and hydroxynaphthoic acid or a salt thereof, and subsequently removing the organic solvent,
(19) a method of producing the sustained-release composition according to term (18) above, wherein the bioactive substance or salt thereof is in the form of an aqueous solution,
(20) a production method according to term (17) above, wherein the salt of the bioactive substance is a salt with a free base or acid,
(21) a pharmaceutical containing the sustained-release composition according to term (1) above,
(22) an agent for preventing or treating prostatic cancer, prostatic hypertrophy, endometriosis, hysteromyoma, metrofibroma, precocious puberty, dysmenorrhea, or breast cancer, or a contraceptive, containing the sustained-release composition according to term (3) above,
(23) a sustained-release composition containing the hydroxynaphthoate of a bioactive substance and a biodegradable polymer or salt thereof,
(24) a method of suppressing bioactive substance initial burst from a sustained-release composition, comprising using hydroxynaphthoic acid or a salt thereof,
(25) a method of increasing the efficiency of bioactive substance inclusion in a sustained-release composition, comprising using hydroxynaphthoic acid or a salt thereof,
(26) a hydroxynaphthoate of a bioactive peptide,
(27) a hydroxynaphthoate of a bioactive peptide according to term (26) above, which is soluble in water or very slightly soluble in water, and
(28) a sustained-release composition containing the hydroxynaphthoate of a bioactive peptide.
The present invention further provides:
(29) a sustained-release composition according to term (28) above, wherein the content of the hydroxynaphthoic acid or salt thereof is about 1 to about 7 mol, preferably about 1 to about 2 mol, per mol of the bioactive peptide or salt thereof
(30) a production method for the sustained-release composition according to term (17) above, comprising producing a W/O emulsion with a solution containing a bioactive substance or salt thereof as an internal aqueous phase and a solution containing a biodegradable polymer and hydroxynaphthoic acid or a salt thereof as an oil phase, and subsequently removing the solvent,
(31) a production method for the sustained-release composition according to term (17) above, comprising producing a W/O emulsion with a solution containing hydroxynaphthoic acid or a salt thereof as an internal aqueous phase and a solution containing a bioactive substance or salt thereof and a biodegradable polymer or salt thereof as an oil phase, and subsequently removing the solvent,
(32) a production method for the sustained-release composition according to term (28) above, comprising mixing and dissolving a bioactive peptide or salt thereof and hydroxynaphthoic acid or salt thereof, and subsequently removing the solvent, and
(33) the production method for sustained-release composition according to any one of terms (30) through (32), wherein the solvent removal method is water-in drying method.
The biologically active substance used in the present invention is not subject to limitation, as long as it is pharmacologically useful, and it may be a non-peptide substance or a peptide substance. The non-peptide substance includes an agonist, an antagonist, and a substance having an enzyme inhibitory activity. The peptide substance includes, for example, biologically active peptides, and particularly those having molecular weights of about 300 to about 40,000, preferably about 400 to about 30,000, and more preferably about 500 to about 20,000.
Such biologically active peptides include, for example, luteinizing hormone-releasing hormone (LH-RH), insulin, somatostatin, growth hormones, growth hormone-releasing hormone (GH-RH), prolactin, erythropoietin, adrenocorticotropic hormone, melanocyte-stimulating hormone, thyroid hormone-releasing hormone, thyroid-stimulating hormone, luteinizing hormone, follicle-stimulating hormone, vasopressin, oxytocin, calcitonin, gastrin, secretin, pancreozymin, cholecystokinin, angiotensin, human placental lactogen, human chorionic gonadotropin, enkephalin, endorphin, kyotorphin, tuftsin, thymopoletin, thymosin, thymostimulin, thymic humoral factor, blood thymic factor, tumor necrosis factor, colony-stimulating factor, motilin, daynorphin, bombesin, neurotensin, caerulein, bradykinin, atrial natriuresis-increasing factor, nerve growth factor, cell growth factor, neurotrophic factor, endothelin-antagonistic peptides, derivatives thereof, fragments of these peptides, and derivatives of such fragments.
The biologically active peptide used in the present invention may be as is, or may be a pharmacologically acceptable salt.
Such salts include salts with inorganic acids (which may also be referred to as inorganic free acids) (e.g., carbonic acid, bicarbonic acid, hydrochloric acid, sulfuric acid, nitric acid, boric acid), organic acids (which may also be referred to as organic free acids)(e.g., succinic acid, acetic acid, propionic acid, trifluoroacetic acid) etc., when said biologically active peptide has a basic group such as an amino group.
When said biologically active peptide has an acidic group such as a carboxyl group, such salts include salts with inorganic bases (which may also be referred to as inorganic free bases) (e.g., alkali metals such as sodium and potassium, alkaline earth metals such as calcium and magnesium), organic bases (which may also be referred to as organic free bases) (e.g., organic amines such as triethylamine, basic amino acids such as arginine) etc. The biologically active peptide may form a metal complex compound (e.g., copper complex, zinc complex).
Preferred examples of the above-described biologically active peptide are LH-RH derivatives or salts thereof that are effective against sex hormone-dependent diseases such as prostatic cancer, prostatic hypertrophy, endometriosis, hysteromyoma, precocious puberty and breast cancer, and effective for contraception.
Examples of LH-RH derivatives or salts thereof include, for example, the peptides described in xe2x80x9cTreatment with GnRH Analogs: Controversies and Perspectivesxe2x80x9d (The Parthenon Publishing Group Ltd., published 1996), Japanese Patent Examined Publication No. 503165/1991, Japanese Patent Unexamined Publication Nos. 101695/1991, 97334/1995 and 259460/1996, and elsewhere.
LH-RH derivatives may be LH-RH agonists or LH-RH antagonists; useful LH-RH antagonists include, for example, biologically active peptides represented by general formula [I]:
X-D2Nal-D4ClPhe-D3Pal-Ser-A-B-Leu-C-Pro-DAlaNH2 (SEQ ID NO.: 2)
[X represents N(4H2-furoyl)Gly or NAc; A represents a residue selected from NMeTyr, Tyr, Aph(Atz) and NMeAph(Atz); B represents a residue selected from DLys(Nic), DCit, DLys(AzaglyNic), DLys(AzaglyFur), DhArg(Et2), DAph(Atz) and DhCi; C represents Lys(Nisp), Arg or hArg(Et2)] or salts thereof.
Useful LH-RH agonists include, for example, biologically active peptides represented by general formula [II]:
5-oxo-Pro-His-Trp-Ser-Tyr-Y-Leu-Arg-Pro-Z (SEQ ID NO.: 1)
[Y represents a residue selected from DLeu, DAla, DTrp, DSer(tBu), D2Nal and DHis(lmBzl); Z represents NH2xe2x80x94C2H5, Gly-NH2] or salts thereof. Peptides wherein Y is DLeu and Z is NHxe2x80x94C2H5, (i.e., a peptide represented by the formula: 5-oxo-Pro-His-Trp-Ser-Tyr-DLeu-Arg-Pro-NHxe2x80x94C2H5 (SEQ ID NO.:3)) in particular, are preferred.
These peptides can be produced by the methods described in the above-mentioned references or patent publications, or methods based thereon.
The abbreviations used herein are defined as follows:
The abbreviations for amino acids are based on abbreviations specified by the IUPAC-IUB Commission on Biochemical Nomenclature [European Journal of Biochemistry, Vol. 138, pp. 9-37 (1984)] or abbreviations in common use in relevant fields. When an optical isomer may be present in amino acid, it is of the L-configuration, unless otherwise stated.
The hydroxynaphthoic acid for the present invention consists of a naphthalene ring and 1 hydroxyl group and 1 carboxyl group, both groups binding to different carbons of the ring. There are therefore a total of 14 isomers with the hydroxyl group located at different positions with respect to the carboxyl group located at positions 1 and 2 of the naphthalene ring. Any of these isomers can be used, and their mixtures in any ratios can be used. As described later, it is preferable that the acid dissociation constant be great, or pKa (pKa=xe2x88x92log10Ka, Ka represents acid dissociation constant) be small. Preference is also given to isomers that are very slightly soluble in water.
Isomers that are soluble in alcohols (e.g., ethanol, methanol) are preferred. The term xe2x80x9csoluble in alcohols,xe2x80x9d as used herein, means that the solubility is not less than 10 g/l in methanol, for example.
Regarding the pKa values of the above-described hydroxynaphthoic acid isomers, the only known value is for 3-hydroxy-2-naphthoic acid (pKa=2.708, Kagaku Binran Kisohen II, Chemical Society of Japan, published Sep. 25, 1969; however, useful information is obtained by comparing the pKa values of three isomers of hydroxybenzoic acid. Specifically, the pKa values of m-hydroxybenzoic acid and p-hydroxybenzoic acid are not less than 4, whereas the pKa value of o-hydroxybenzoic acid (salicylic acid) (=2.754) is extremely small. Of the above-mentioned 14 isomers, those consisting of a naphthalene ring and a carboxyl group and a hydroxyl group, both bound to adjoining carbon atoms of the ring, i.e., 3-hydroxy-2-naphthoic acid, 1-hydroxy-2-naphthoic acid, and 2-hydroxy-1-naphthoic acid are therefore preferred. Furthermore, 3-hydroxy-2-naphthoic acid, which consists of a naphthalene ring and a hydroxyl group bound to the carbon at position 3 of the ring and 1 carboxyl group bound to the carbon at position 2 of the ring, is preferred.
The hydroxynaphthoic acid may be a salt. Salts include, for example, salts with inorganic bases (e.g., alkali metals such as sodium and potassium, alkaline earth metals such as calcium and magnesium), organic bases (e.g., organic amines such as triethylamine, basic amino acids such as arginine), and salts and complex salts with transition metals (e.g., zinc, iron, copper).
An example method of preparing the hydroxynaphthoic acid salt of the bioactive substance of the present invention is given below.
(1) A hydrated organic solvent solution of hydroxynaphthoic acid is passed through a weakly basic ion exchange column to adsorb the acid and saturate the column. The excess portion of the hydroxynaphthoic acid is then removed through the hydrated organic solvent, after which a hydrated organic solvent solution of the bioactive substance or salt thereof is passed through the column to cause ion exchange; the solvent is removed from the effluent obtained. Useful organic solvents in said hydrated organic solvent include alcohols (e.g., methanol, ethanol), acetonitrile, tetrahydrofuran, and dimethylformamide. Solvent removal for salt precipitation is achieved using a commonly known method or a method based thereon. Examples of such methods include the method in which the solvent is evaporated, with the degree of vacuum adjusted using a rotary evaporator etc.
(2) Through a weakly basic ion exchange column, previously subjected to ion exchange to hydroxide ions, a hydrated organic solvent solution of the bioactive substance or salt thereof is passed, to convert the basic groups to the hydroxide type. Hydroxynaphthoic acid in an amount not more than the molar equivalent is added to the effluent recovered, and dissolved, followed by concentration; the salt precipitated is washed with water as necessary, and dried.
Because the hydroxynaphthoic acid salt of a bioactive substance is very slightly soluble in water, although also depending on the bioactive substance used, said salt itself of a bioactive peptide, exhibiting potential for sustained-release, can be used for a sustained-release preparation of a bioactive substance, and can also be used to produce a sustained-release composition.
Biodegradable polymers used in the present invention include, for example, polymers and copolymers that have been synthesized from one or more kinds selected from xcex1-hydroxymonocarboxylic acids (e.g., glycolic acid, lactic acid), hydroxydicarboxylic acids (e.g., malic acid), hydroxytricarboxylic acids (e.g., citric acid) etc., and that have a free carboxyl group, or mixtures thereof; poly-xcex1-cyanoacrylic acid esters; polyamino acids (e.g., poly-g-benzyl-L-glutamic acid); and maleic anhydride copolymers (e.g., styrene-maleic acid copolymers).
The mode of monomer binding may be random, block, or graft. When the above-mentioned xcex1-hydroxymonocarboxylic acids, xcex1-hydroxydicarboxylic acids, and xcex1-hydroxytricarboxylic acids have an optically active center in their molecular structures, they may be of the D-, L- or DL-configuration. Of these, lactic acid-glycolic acid polymers [hereinafter also referred to as poly(lactide-co-glycolide), poly(lactic acid-co-glycolic acid) or lactic acid-glycolic acid copolymer; generically refer to lactic acid-glycolic acid homopolymers and copolymers, unless otherwise specified; lactic acid homopolymers are also referred to as lactic acid polymer, polylactic acids, polylactides etc., and glycolic acid homopolymers as glycolic acid polymers, polyglycolic acids, polyglycolides etc.], with preference given to poly(xcex1-cyanoacrylic esters) etc. Greater preference is given to lactic acid-glycolic acid polymers. More preferably, lactic acid-glycolic acid polymers having a free carboxyl group at one end are used.
The biodegradable polymer may be a salt. Salts include, for example, salts with inorganic bases (e.g., alkali metals such as sodium and potassium, alkaline earth metals such as calcium and magnesium), organic bases. (e.g., organic amines such as triethylamine, basic amino acids such as arginine), and salts and complex salts with transition metals (e.g., zinc, iron, copper).
When the biodegradable polymer used is a lactic acid-glycolic acid polymer, the content ratio (mol %) is preferably about 100/0 to about 40/60, more preferably about 100/0 to about 50/50. Lactic acid homopolymers having a content ratio of 100/0 are also preferably used.
The optical isomer ratio of lactic acid, one of the minimum repeat units for said lactic acid-glycolic acid polymer is preferably between about 75/25 and about 25/75, as of the D-configuration/L-configuration ratio (mol/mol %). Lactic acid-glycolic acid polymers having a D-configuration/L-configuration ratio (mol/mol %) between about 60/40 and about 30/70 are commonly used.
The weight-average molecular weight of said lactic acid-glycolic acid polymer is normally about 3,000 to about 100,000, preferably about 3,000 to about 60,000, more preferably about 3,000 to about 50,000, and still more preferably about 20,000 to about 50,000.
The degree of dispersion (weight-average molecular weight/number-average molecular weight) is normally about 1.2 to about 4.0, more preferably about 1.5 to 3.5.
The free carboxyl group content of said lactic acid-glycolic acid polymer is preferably about 20 to about 1,000 xcexcmol, more preferably about 40 to about 1,000 xcexcmol, per unit mass (gram) of the polymer.
Weight-average molecular weight, number-average molecular weight and degree of dispersion, as defined herein, are polystyrene-based molecular weights and degree of dispersion determined by gel permeation chromatography (GPC) with 15 polystyrenes as reference substances with weight-average molecular weights of 1,110,000, 707,000, 455,645, 354,000, 189,000, 156,055, 98,900, 66,437, 37,200, 17,100, 9,830, 5,870, 2,500, 1,303, and 504, respectively. Measurements were taken using a high-speed GPC device (produced by Toso, HLC-8120GPC, detection:Refractory Index) and a GPC column KF804Lx2 (produced by Showa Denko), with chloroform as a mobile phase.
The term free carboxyl group content, as used herein, is defined to be obtained by the labeling method (hereinafter referred to as xe2x80x9ccarboxyl group content as determined by the labeling methodxe2x80x9d). Specific procedures for determining this content in a polylactic acid are described below. First, W mg of the polylactic acid is dissolved in 2 ml of a 5 N hydrochloric acid/acetonitrile (v/v=4/96) mixture; 2 ml of a 0.01 M solution of o-nitrophenylhydrazine hydrochloride (ONPH) (5 N hydrochloric acid/acetonitrile/ethanol=1.02/35/15) and 2 ml of a 0.15 M solution of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (pyridine/ethanol=4v/96v) were added, followed by a reaction at 40xc2x0 C. for 30 minutes, after which the solvent is removed. After water washing (4 times), the residue is dissolved in 2 ml of acetonitrile; 1 ml of a 0.5 mol/l ethanolic solution of potassium hydroxide is added, followed by a reaction at 60xc2x0 C. for 30 minutes. The reaction mixture is diluted with a 1.5 N aqueous solution of sodium hydroxide to Y ml; absorbance a (/cm) at 544 nm is determined, with a 1.5 N aqueous solution of sodium hydroxide as control. Separately, with an aqueous solution of DL-lactic acid as reference, its free carboxyl group content C mol/l is determined by alkali titration. Taking the absorbance at 544 nm of the DL-lactic acid hydrazide prepared by the ONPH labeling method as B (/cm), the molar content of the free carboxyl groups per unit mass (gram) of the polymer can be calculated using the equation:
[COOH](mol/g)=(AYC)/(WB)
Although said carboxyl group content can also be obtained by dissolving the biodegradable polymer in a toluene-acetone-methanol mixed solvent, and titrating this solution for carboxyl groups with an alcoholic solution of potassium hydroxide, with phenolphthalein as indicator (value obtained by this method hereinafter referred to as xe2x80x9ccarboxyl group content as determined by the alkali titration methodxe2x80x9d), it is desirable that quantitation be achieved by the labeling method described above, since it is possible that the titration endpoint is made unclear as a result of competition of the hydrolytic reaction of the polyester main chain during titration.
The decomposition/elimination rate of a biodegradable polymer varies widely, depending on copolymer composition, molecular weight or free carboxyl group content. However, drug release duration can be extended by lowering the glycolic acid ratio or increasing the molecular weight and lowering the free carboxyl group content, because decomposition/elimination is usually delayed as the glycolic acid ratio decreases, in the case of lactic acid-glycolic acid polymers. Because the free carboxyl group content affects the rate of bioactive substance incorporation in the preparation, however, it must be above a given level. For this reason, it is preferable, in obtaining a biodegradable polymer for a sustained-release preparation of the long acting type (e.g., 6 months or longer), that in the case of a lactic acid-glycolic acid polymer, a polylactic acid (e.g., D-lactic acid, L-lactic acid, DL-lactic acid, preferably DL-lactic acid etc.) whose weight-average molecular weight and free carboxyl group content as determined as described above are about 20,000 to about 50,000 and about 30 to about 95 xcexcmol/g, preferably about 40 to 30 about 95)xcexcmol/g, more preferably about 50 to about 90 xcexcmol/g, be used.
Said xe2x80x9clactic acid-glycolic acid polymerxe2x80x9d can be produced by, for example, the catalyst-free dehydration polymerization condensation method (Japanese Patent Unexamined Publication No. 28521/1986) from a lactic acid and a glycolic acid, or ring-opening polymerization from a lactide and a cyclic diester compound such as glycolide by means of a catalyst (Encyclopedic Handbook of Biomaterials and Bioengineering Part A: Materials; Volume 2, Marcel Dekker, Inc., 1995). Although the polymer obtained by the above-mentioned known method of ring-opening polymerization does not always contain a free carboxyl group at one end, it can also be used after being modified to a polymer having a given amount of carboxyl groups per unit mass, by subjecting it to the hydrolytic reaction described in EP-A-0839525.
The above-described xe2x80x9clactic acid-glycolic acid polymer having a free carboxyl group at one endxe2x80x9d can be produced, with no problem, by a commonly known method (e.g., catalyst-free dehydration polymerization condensation, Japanese Patent Unexamined Publication No. 28521/1986), or by the method described below.
(1) First, in the presence of a hydroxymonocarboxylic acid derivative (e.g., tert-butyl D-lactate, benzyl L-lactate) with its carboxyl group protected, or a hydroxydicarboxylic acid derivative (e.g., dibenzyl tartronate, di-tert-butyl 2-hydroxyethylmalonate) with its carboxyl group protected, a cyclic ester compound is subjected to a polymerization reaction using a polymerization catalyst.
The above-described xe2x80x9chydroxymonocarboxylic acid derivative with its carboxyl group protectedxe2x80x9d or xe2x80x9chydroxydicarboxylic acid derivative with its carboxyl group protectedxe2x80x9d is exemplified by hydroxycarboxylic acid derivatives with its carboxyl group (xe2x80x94COOH) amidated (xe2x80x94CONH2) or esterified (xe2x80x94COOR), with preference given to hydroxycarboxylic acid derivatives with its carboxyl group (xe2x80x94COOH) esterified (xe2x80x94COOR) etc.
Here, R for the ester is exemplified by C1-6 alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, and tert-butyl, C3-8 cycloalkyl groups such as cyclopentyl and cyclohexyl, C6-12 aryl groups such as phenyl and xcex1-naphthyl, and C7-14 aralkyl groups such as phenyl-C1-2 alkyl groups such as benzyl and phenethyl, and xcex1-naphthyl-C1-2 alkyl groups such as xcex1-naphthylmethyl. Of these groups, tert-butyl groups, benzyl groups etc. are preferred.
Said xe2x80x9ccyclic ester compoundxe2x80x9d refers to a cyclic compound having at least one ester linkage in the ring thereof. Specifically, such compounds include cyclic monoester compounds (lactones) or cyclic diester compounds (lactides).
Said xe2x80x9ccyclic monoester compoundxe2x80x9d is exemplified by 4-membered ring lactones (xcex2-propiolactone,xcex2-butyrolactone, xcex2-isovalerolactone, xcex2-caprolactone, xcex2-isocaprolactone, xcex2-methyl-xcex2-valerolactone etc.), 5-membered ring lactones (xcex3-butyrolactone, xcex3-valerolactone etc.), 6-membered ring lactones (xcex4-valerolactone etc.), 7-membered ring lactones (xcex5-caprolactone etc.), p-dioxanone, and 1,5-dioxepan-2-one.
Said xe2x80x9ccyclic diester compoundxe2x80x9d is exemplified by the compounds represented by the formula:
Wherein R1 and R2, whether identical or not, represent a hydrogen atom or a C1-6 alkyl group such as methyl, ethyl, n-propyl, isopropyl, n-butyl, or tert-butyl), with preference given to the lactides having a hydrogen atom for R1 and a methyl group for R2, or having a hydrogen atom for R1 and R2, etc.
Specifically, such compounds include glycolides, L-lactides, D-lactides, DL-lactides, meso-lactides, and 3-methyl-1,4-dioxane-2,5-dione (including optically active configurations).
Said xe2x80x9cpolymerization catalystxe2x80x9d is exemplified by organic tin catalysts (e.g., tin octylate, di-n-butyltin dilaurylate, tetraphenyltin), aluminum catalysts (e.g., triethylaluminum), and zinc catalysts (e.g., diethylzinc).
From the viewpoint of ease of removal after reaction, aluminum catalysts and zinc catalysts are preferred; from the viewpoint of safety In case of retention, zinc catalysts are preferred.
Useful solvents for polymerization catalysts include benzene, hexane, and toluene, with preference given to hexane, toluene etc.
Regarding xe2x80x9cmethod of polymerization,xe2x80x9d the mass polymerization method, which is conducted with the reaction product in a molten state, or the solution polymerization method, which is conducted with the reaction product dissolved in an appropriate solvent (e.g., benzene, toluene, xylene, decalin, dimethylformamide). Although polymerization temperature is not limited, it is not lower than the temperature at which the reaction product becomes molten at reaction initiation, normally 100 to 300xc2x0 C., for mass polymerization, and is normally room temperature to 150xc2x0 C. for solution polymerization; if the reaction temperature exceeds the boiling point of the reaction solution, the reaction is carried out under refluxing using a condenser or in a pressure-resistant reactor. Determined as appropriate in consideration of polymerization temperature, other reaction conditions, physical properties of the desired polymer, etc., polymerization time is, for example, 10 minutes to 72 hours. After completion of the reaction, polymerization is terminated with an acid (e.g., hydrochloric acid, acetic anhydride, trifluoroacetic acid), with the reaction mixture dissolved in an appropriate solvent (e.g., acetone, dichloromethane, chloroform) if necessary, after which the mixture is mixed in a solvent that does not dissolve the desired product (e.g., alcohol, water, ether, isopropyl ether) or otherwise precipitated, followed by the isolation of a polymer having a protected carboxyl group at the xcfx89-end.
The method of polymerization of the present application employs hydroxycarboxylic acid derivatives (e.g., tert-butyl D-lactate, benzyl L-lactate) with a protected carboxyl group or hydroxydicarboxylic acid derivatives (e.g., dibenzyl tartronate, di-tert-butyl L-2-hydroxyethylmalonate) with a protected carboxyl, in place of conventional protonic chain transferring agents such as methanol.
Using hydroxycarboxylic acid derivatives (e.g., tert-butyl D-lactate, benzyl L-lactate) with a protected carboxyl group or hydroxydicarboxylic acid derivatives (e.g., dibenzyl tartronate, di-tert-butylL-2-hydroxyethylmalonate) with a protected carboxyl, as protonic chain transferring agents, it is possible to {circle around (1)} achieve molecular weight control by the composition of the starting materials, and to {circle around (2)} liberate the carboxyl group at the xcfx89-end of the biodegradable polymer obtained, by a deprotection reaction after polymerization.
(2) Second, by subjecting the polymer obtained by the polymerization reaction described in paragraph (1) above, which has a protected carboxyl group at the xcfx89-end, to a deprotection reaction, the desired biodegradable polymer having a free carboxyl group at the xcfx89-end can be obtained.
Said protecting group can be removed by commonly known methods. Such methods include all methods enabling the removal of the protecting group without affecting the ester linkage of poly(hydroxycarboxylic acid), specifically exemplified by reduction and acid decomposition.
Such methods of reduction include, for example, catalytic reduction using catalysts (e.g., palladium carbon, palladium black, platinum oxide), reduction using sodium in liquid ammonium, and reduction with dithiothreitol. When a polymer having a protected carboxyl group at the xcfx89-end is subjected to catalytic reduction, for example, deprotection can be achieved by adding palladium carbon to a solution of the polymer in ethyl acetate, dichloromethane, chloroform, or the like, and supplying hydrogen at room temperature for about 20 minutes to about 4 hours with vigorous shaking.
Such methods of acid decomposition include, for example, acid decomposition with inorganic acids (e.g., hydrogen fluoride, hydrogen bromide, hydrogen chloride), organic acids (e.g., trifluoroacetic acid, methanesulfonic acid, trifluoromethanesulfonic acid), or mixtures thereof. Also, where necessary, a cation scavenger (e.g., anisole, phenol, thioanisole) is added as appropriate. When a polymer having a carboxyl group protected with a tert-butyl group, at the xcfx89-end, is subjected to acid decomposition, for example, deprotection can be achieved by adding an appropriate amount of trifluoroacetic acid to a solution of the polymer in dichloromethane, xylene, toluene, or the like, or dissolving the polymer in trifluoroacetic acid, and stirring at room temperature for about 1 hour.
Preferably, said acid decomposition is conducted just after the polymerization reaction; in this case, it can serve as a polymerization termination reaction.
Furthermore, by subjecting the polymer obtained by the above-described deprotection reaction to an acid hydrolytic reaction as necessary, the weight-average molecular weight, number-average molecular weight or terminal carboxyl group content of said polymer can be regulated according to the purpose. Specifically, this can, for example, be achieved by the method described in EP-A-0839525 or a method based thereon.
A biodegradable polymer obtained as described above can be used as a base for producing a sustained-release preparation.
In addition, a polymer having a given free carboxyl group at one end can be produced by known production methods (e.g., see Patent Publication for WO94/15587).
Also, a lactic acid-glycolic acid polymer with one end rendered a free carboxyl group by a chemical treatment after ring-opening polymerization may be a commercial product of Boehringer Ingelheim KG, for example.
The biodegradable polymer may be a salt (salts of biodegradable polymers include, for example, the salts mentioned above). Useful methods of their production include, for example, (a) the method in which a solution of the above-described biodegradable polymer having a carboxyl group in an organic solvent, and an aqueous solution containing the ions of inorganic bases (e.g., alkali metals such as sodium and potassium, alkaline earth metals such as calcium and magnesium) or organic bases (e.g., organic amines such as triethylamine, basic amino acids such as arginine), are mixed together to cause an ion exchange reaction, after which the polymer, now in the form of a salt, is isolated, (b) the method in which a weak acid salt (e.g. acetate, glycolate) of a base listed in (a) above is dissolved in a solution of the above-described biodegradable polymer having a carboxyl group in an organic solvent, after which the polymer, now in the form of a salt, is isolated, and (c) the method in which a weak acid salt (e.g. acetate, glycolate) or oxide of a transition metal (e.g., zinc, iron, copper) is mixed in a solution of the above-described biodegradable polymer having a carboxyl group in an organic solvent, after which the polymer, now in the form of a salt, is isolated.
As a biodegradable polymer for a sustained-release preparation of the long acting type (e.g., 6 months or longer), the xe2x80x9clactic acid-glycolic acid polymer having a free carboxyl group at one endxe2x80x9d produced by the method described above is preferred.
The ratio by weight of the bioactive substance in the composition of the present invention varies depending on kind of bioactive substance, desired pharmacological effect, duration of effect, and other factors. In the case of a sustained-release composition containing three components (bioactive substance or salt thereof, hydroxynaphthoic acid or salt thereof, and biodegradable polymer or salt thereof), the ratio by weight of bioactive peptide or salt thereof, for example, is about 0.001 to about 50% by weight, preferably about 0.02 to about 40% by weight, more preferably about 0.1 to 30% by weight, and most preferably about 14 to 24% by weight, relative to the sum of the three components. In the case of a non-peptide bioactive substance or salt thereof, the ratio is about 0.01 to 80% by weight, preferably about 0.1 to 50% by weight. When the hydroxynaphthoic acid salt of a bioactive substance is contained, similar ratios by weight are applicable. In the case of a sustained-release composition containing the salt of a bioactive peptide (referred to as (A)) with hydroxynaphthoic acid (referred to as (B)), the ratio by weight of (A) is normally about 5 to about 90% by weight, preferably about 10 to about 85% by weight, more preferably about 15 to about 80% by weight, and still more preferably about 30 to about 80% by weight, relative to the sum of the salt (A) with (B).
In the case of a sustained-release composition containing three components (bioactive substance or salt thereof, hydroxynaphthoic acid or salt thereof, and biodegradable polymer or salt thereof), the amount of hydroxynaphthoic acid or salt thereof formulated is preferably about 1/2 to about 2 mol, more preferably about 3/4 to about 4/3 mol, and still more preferably about 4/5 to about 6/5 mol, per mol of bioactive substance or salt thereof.
Designing the composition of the present invention is hereinafter described for a sustained-release composition containing three components: basic bioactive substance, hydroxynaphthoic acid, and biodegradable polymer. In this case, the bioactive substance, as a base, and hydroxynaphthoic acid, as an acid, are concurrently present in the composition; whether they are formulated in the composition in the form of free configurations or salts, a dissociation equilibrium for each component is present, in a hydrated state, or in the presence of a trace amount of water, at a point during production of the composition. Because the salt formed by any hydroxynaphthoic acid, which is very slightly soluble in water, with a bioactive substance is assumed to be very slightly soluble in water, although the solubility also depends on the characteristics of said bioactive substance, its dissociation equilibrium shifts toward the formation of a salt very slightly soluble in water.
In producing a composition having high contents of a basic bioactive substance, it is desirable that most of the bioactive substance be protonated to render it to a salt very slightly soluble in water as described above, judging from the above-described dissociation equilibrium. For this purpose, it is desirable that the hydroxynaphthoic acid or salt thereof be formulated in an amount at least nearly equivalent to that of the bioactive substance or salt thereof.
Next, the mechanism of release of a bioactive substance included in a composition is described below. In the above-described formula composition, the bioactive substance is mostly protonated and present with a counter ion. The counter ton is mainly hydroxynaphthoic acid (preferably hydroxynaphthoic acid). After the composition is administered to the living body, its oligomers and monomers begin to be produced over time due to decomposition of the biodegradable polymer. When said polymer is a lactic acid-glycolic acid polymer, the resulting oligomer (lactic acid-glycolic acid oligomer) and monomer (lactic acid or glycolic acid) always has one carboxyl group, which can also serve as a counter ion for the bioactive substance. The bioactive substance is released without charge transfer, or in the form of a salt with a counter ion; transferable counter ions include hydroxynaphthoic acids, lactic acid-glycolic acid oligomers (of such molecular weights that transfer is possible), and monomers (lactic acid or glycolic acid), as described above.
When a plurality of acids are concurrently present, salts of stronger acids are usually preferentially produced, although the outcome also depends on their content ratio. Regarding the pKa values of hydroxynaphthoic acids, 3-hydroxy-2-naphthoic acid, for example, is known to have a pKa value of 2.708 (Kagaku Binran Kisohen II, Chemical Society of Japan, published Sep. 25, 1969). On the other hand, the pKa values of the carboxyl groups of lactic acid-glycolic acid oligomers are unknown but can be calculated on the basis of the pKa value of lactic acid or glycolic acid (=3.86 or 3.83), in accordance with the theory that xe2x80x9cthe free energy level change due to substituent introduction can be approximated by the addition rule.xe2x80x9d The contributions of substituents to dissociation constants have already been determined and can be used for this purpose (Table 4.1 in xe2x80x9cpKa Prediction for Organic Acid and Bases,xe2x80x9d D. D. Perrin, B. Dempsey, and E. P. Serjeant, 1981). Because the following data are applicable for the hydroxyl group and ester linkage:
xcex94pKa (OH)=xe2x88x920.90
xcex94pKa (ester linkage)=xe2x88x921.7
the pKa value of the carboxyl group of lactic acid-glycolic acid oligomers can be determined, in consideration of the contribution of the ester linkage closest to the dissociation group, as follows:
pKa=pKa (lactic acid or glycolic acid)xe2x88x92xcex94pKa (OH)+xcex94pKa (ester linkage)=3.06 or 3.03
Because hydroxynaphthoic acids are therefore stronger acids than lactic acid (pKa=3.86.), glycolic acid (pKa=3.83), and lactic acid-glycolic acid oligomers, it is assumed that the hydroxynaphthoic acid salt of the bioactive substance is preferentially produced in the above-described composition, the characteristics of the salt being assumed to predominantly determine the nature of sustained-release of the bioactive substance from the composition. Said bioactive substance is exemplified by the bioactive substances described above.
Here, the fact that the salt formed by the hydroxynaphthoic acid with the bioactive substance is very slightly soluble in water, rather than insoluble in water, serves in favor of the sustained-release mechanism. In other words, as demonstrated in the above discussion of acid dissociation constant, the salt of hydroxynaphthoic acid, a stronger acid than the above-described lactic acid-glycolic acid oligomers and monomers, is predominant in the initial stage of release; the initial release pattern of the drug can be regulated by the content ratio of hydroxynaphthoic acid, because the solubility and body tissue distribution profile of the salt serves as determinants of the bioactive substance release rate. Then, as the oligomers and monomers increase, due to reduction in the hydroxynaphthoic acid and hydrolysis of the biodegradable polymer, the bioactive substance release mechanism involving oligomers and monomers as counter ions becomes predominant gradually; even if the hydroxynaphthoic acid disappears substantially from said xe2x80x9ccomposition,xe2x80x9d stable bioactive substance release is achieved. The increased efficiency of bioactive substance incorporation for production of a sustained-release composition, and the possibility of suppression of initial burst after administration of the bioactive substance incorporated, can also be explained.
The role of the hydroxynaphthoic acid in the sustained-release composition containing the hydroxynaphthoic acid salt of a bioactive peptide can also be explained by the above-described mechanism.
The term xe2x80x9cinsoluble in water,xe2x80x9d as used herein, means that when said substance is stirred in distilled water for 4 hours at temperatures of not higher than 40xc2x0 C., the mass of the substance that dissolves in 1 l of the solution is not more than 25 mg.
The term xe2x80x9cvery slightly soluble in water,xe2x80x9d as used herein, means that the above-described mass is not less than 25 mg and not more than 5 g. When said substance is a salt of a bioactive substance, the above definition is applied for the mass of the bioactive substance that dissolved in the above-described operation.
Although the sustained-release composition of the present invention is not subject to limitation as to form, microparticles are preferred, with greater preference given to microspheres (also referred to as microcapsules in the case of sustained-release compositions containing biodegradable polymers). The term xe2x80x9cmicrosphere,xe2x80x9d as used herein, is defined as an injectable sphere that can be dispersed in solutions. Its shape can be confirmed by, for example, scanning microscopy.
Production methods for sustained-release compositions of the present invention, which contain a biologically active substance or a salt thereof, a hydroxynaphthoic acid or a salt thereof, and a biodegradable polymer or a salt thereof, microspheres, are exemplified below.
(i) O/W Method
In this method, an organic solvent solution of the hydroxynaphthoic acid or a salt thereof and biodegradable polymer or a salt thereof is prepared.
Said organic solvent is exemplified by halogenated hydrocarbons (e.g., dichloromethane, chloroform, dichloroethane, trichloroethane, carbon tetrachloride), ethers (e.g., ethyl ether, isopropyl ether), fatty acid esters (e.g., ethyl acetate, butyl acetate), aromatic hydrocarbons (e.g., benzene, toluene, xylene), alcohols (e.g., ethanol, methanol), and acetonitrile. Among these, dichloromethane is preferable for an organic solvent of the biodegradable polymer or a salt thereof. Alcohols are preferable for an organic solvent of the hydroxynaphthoic acid or a salt thereof. These solvents may be used in mixtures at appropriate ratios. Of these solvents, mixtures of halogenated hydrocarbons and alcohols are preferred, with greater preference given to mixtures of dichloromethane and ethanol.
When the organic solvent used is a mixture of dichloromethane and ethanol, the ratio of their concentrations is normally chosen over the range from about 0.01 to about 50% (v/v), preferably from about 0.05 to about 40% (v/v), and more preferably from about 0.1 to about 30% (v/v).
The biodegradable polymer concentration in the organic solvent solution varies depending on the molecular weight of biodegradable polymer and the kind of organic solvent. For example, when the organic solvent used is dichloromethane, the biodegradable polymer concentration is normally chosen over the range from about 0.5 to about 70% by weight, preferably from about 1 to about 60% by weight, and more preferably from about 2 to about 50% by weight.
The hydroxynaphthoic acid or a salt thereof concentration in the organic solvent solution is normally chosen, for example, over the range from about 0.01 to about 10% by weight, preferably from about 0.1 to about 5% by weight, and more preferably from about 0.5 to about 3% by weight.
The biologically active substance or salt thereof is added to thus-obtained organic solvent solution containing a hydroxynaphthoic acid or salt thereof, and a biodegradable polymer, and dissolved or dispersed.
The thus-obtained organic solvent solution containing a biologically active substance or salt thereof, a hydroxynaphthoic acid or salt thereof, and a biodegradable polymer, is then added to a water phase to form an O (oil phase)/W (water phase) emulsion, after which the solvent is evaporated from the oil phase to yield microspheres. For this operation, the water phase volume is normally chosen over the range from about 1 time to about 10,000 times, preferably from about 5 times to about 50,000 times, and more preferably from about 10 times to about 2,000 times, the oil phase volume.
An emulsifier may be added to the above-described external water phase. Said emulsifier may be any one, as long as it is capable of forming a stable O/W emulsion. Such emulsifiers include, for example, anionic surfactants (e.g., sodium oleate, sodium stearate, sodium lauryl sulfate), nonionic surfactants [e.g., polyoxyethylene sorbitan fatty acid esters (Tween 80, Tween 60, Atlas Powder Company), polyoxyethylene castor oil derivatives (e.g., HCO-60, HCO-50, Nikko Chemicals)], polyvinylpyrrolidone, polyvinyl alcohol, carboxymethyl cellulose, lecithin, gelatin and hyaluronic acid. These emulsifiers may be used singly or in combination. Regarding the concentration, it is preferable that they be used over the range from about 0.01% to 10% by weight, preferably from about 0.05% to about 5% by weight.
An osmotic pressure regulator may be added to the above-described external water phase. Said osmotic pressure regulator may be any one, as long as it shows an osmotic pressure when prepared as an aqueous solution.
Said osmotic pressure regulator is exemplified by polyhydric alcohols, monohydric alcohols, monosaccharides, disaccharides, oligosaccharides, amino acids, and derivatives thereof.
Useful polyhydric alcohols include, for example, dihydric alcohols such as glycerol, pentahydric alcohols such as arabitol, xylitol and adonitol, and hexahydric alcohols such as mannitol, sorbitol and dulcitol. Of these alcohols, hexavalent alcohols are preferred, with greater preference given to mannitol.
Useful monohydric alcohols include, for example, methanol, ethanol and isopropyl alcohol, with preference given to ethanol.
Useful monosaccharides include, for example, pentoses such as arabinose, xylose, ribose and 2-deoxyribose, and hexoses such as glucose, fructose, galactose, mannose, sorbose, rhamnose and fucose, with preference given to pentoses.
Useful oligosaccharides include, for example, trisaccharides such as maltotriose and raffinose, and tetrasaccharides such as stachyose, with preference given to trisaccharides.
Useful derivatives of monosaccharides, disaccharides and oligosaccharides include, for example, glucosamine, galactosamine, glucuronic acid and galacturonic acid.
Useful amino acids include, for example, glycine, leucine and arginine, with preference given to L-arginine.
These osmotic pressure regulators may be used singly, or in combination.
These osmotic pressure regulators are normally used at such concentrations that the external water phase osmotic pressure is about 1/50 to about 5 times, preferably about 1/25 to about 3 times, the physiological saline osmotic pressure.
Organic solvent removal can be achieved by commonly known methods or methods based thereon. Such methods include, for example, the method in which the organic solvent is evaporated under normal or gradually reduced pressure during stirring using a propeller stirrer, magnetic stirrer or the like, and the method in which the organic solvent is evaporated while the degree of vacuum is adjusted using a rotary evaporator or the like.
The thus-obtained microspheres are centrifuged or filtered to separate them, after which they are washed with distilled water several times to remove the free biologically active substance, hydroxynaphthoic acid, drug support, emulsifier etc. adhering to the microsphere surface, then again dispersed in distilled water etc. and freeze-dried.
To prevent mutual aggregation of particles during the production process, an anticoagulant may be added. Said anticoagulant is exemplified by water-soluble polysaccharides such as mannitol, lactose, glucose and starches (e.g., corn starch), amino acids such as glycine, and proteins such as fibrin and collagen. Of these substances, mannitol is preferred.
Where necessary, freeze-drying may be followed by heating under reduced pressure without causing mutual adhesion of microspheres, to remove the water and organic solvent from the microspheres. It is preferable that the microspheres be heated at a temperature slightly higher than the intermediate glass transition point of the biodegradable polymer, as determined using a differential scanning calorimeter when the temperature is increased at a rate of 10 to 20xc2x0 C. per minute. More preferably, the microspheres are heated within the temperature range from the intermediate glass transition point of the biodegradable polymer to a temperature higher by about 30xc2x0 C. than the glass transition temperature. When a lactic acid-glycolic acid polymer is used as the biodegradable polymer, in particular, it is preferable that the microspheres be heated within the temperature range from the intermediate glass transition point to a temperature higher by 10xc2x0 C. than the glass transition temperature, more preferably within the temperature range from the intermediate glass transition point to a temperature higher by 5xc2x0 C. than the glass transition temperature.
Although it varies depending on the amount of microspheres and other factors, heating time is normally about 12 hours to about 168 hours, preferably about 24 hours to about 120 hours, and more preferably about 48 hours to about 96 hours, after the microspheres reach a given temperature.
Any heating method can be used, as long as microsphere aggregates are uniformly heated.
Useful thermal drying methods include, for example, the method in which thermal drying is conducted in a constant-temperature chamber, fluidized bed chamber, mobile chamber or kiln, and the method using microwaves for thermal drying. Of these methods, the method in which thermal drying is conducted in a constant-temperature chamber is preferred.
(ii). W/O/W Method (1)
First, an organic solvent solution containing a biodegradable polymer or salt thereof is prepared.
The concentration of the organic solvent and the biodegradable polymer or salt thereof are the same as those described in paragraph (I) (i) above.
When more than two kinds of solvents are used, the ratios of these solvents are the same as those described in paragraph (I) (i) above. The biologically active substance or salt thereof is added to thus-obtained organic solvent solution containing the biodegradable polymer, then dissolved and dispersed.
Next, to the organic solvent solution (oil phase) of the biologically active substance and biodegradable polymer, a solution of a hydroxynaphthoic acid or salt thereof [this solvent exemplified by water, alcohols (e.g., methanol, ethanol), pyridine solution, dimethylacetamide solution etc.] is added. This mixture is emulsified by a known method such as homogenization or sonication to form a W/O emulsion.
The thus-obtained W/O emulsion containing a biologically active substance or salt thereof, hydroxynaphthoic acid or salt thereof, and a biodegradable polymer or salt thereof, is then added to a water phase to form a W (internal water phase)/O (oil phase)/W (external water phase) emulsion, after which the solvent is evaporated from the oil phase to yield microspheres. For this operation, the external water phase volume is normally chosen over the range from about 1 time to about 10,000 times, preferably from about 5 times to about 50,000 times, and more preferably from about 10 times to about 2,000 times, the oil phase volume.
The above-described emulsifier and osmotic pressure regulator that may be added to the external water phase, and the subsequent procedures are the same as those described in paragraph (I) (i) above.
ii) W/O/W Method (2)
First, an organic solvent solution containing a hydroxynaphthoic acid and a biodegradable polymer is prepared. Thus-obtained organic solvent solution is referred to as an oil phase. The preparation method is the same as those described in paragraph (I) (i) above.
Alternatively, an organic solvent solution containing a hydroxynaphthoic acid and an organic solvent solution containing a biodegradable polymer may be prepared separately, and mixed together to prepare the oil phase.
The biodegradable polymer concentration in the organic solvent solution varies depending on the molecular weight of biodegradable polymer and the kind of organic solvent. For example, when the organic solvent used is dichloromethane, the biodegradable polymer concentration is normally chosen over the range from about 0.5 to about 70% by weight, preferably from about 1 to about 60% by weight, and more preferably from about 2 to about 50% by weight.
Next, a solution of a biologically active substance or salt thereof [this solvent exemplified by water, alcohols (e.g., methanol, ethanol)] is prepared. Thus-obtained solution is referred to as internal water phase. The concentration of the biologically active substance is normally 0.001 mg/ml to 10 g/ml, preferably, 0.1 mg/ml to 5 g/ml, more preferably, 10 mg/ml to 3 g/ml. The oil phase and the internal water phase are emulsified by a known method such as homogenization or sonication to form a W/O emulsion.
For this operation, the oil phase volume is normally chosen over the range from about 1 time to about 1,000 times, preferably from about 2 times to about 100 times, and more preferably from about 3 times to about 10 times, the internal water phase volume.
The viscosity of the w/o emulsion is normally chosen over the range from about 10 to about 1,0000 cp, preferably from about 100 to about 5,000 cp, more preferably from about 500 to about 2,000 cp.
Thus-obtained W/O emulsion containing a biologically active substance or salt thereof, hydroxynaphthoic acid or salt thereof, and a biodegradable polymer, is then added to a water phase to form a w(internal water phase)/o(oil phase)/w(external water phase) emulsion, after which the solvent is evaporated from the oil phase to yield microspheres. For this operation, the external water phase volume is normally chosen over the range from about 1 time to about 10,000 times, preferably from about 2 times to about 100 times, and more preferably from about 3 times to about 10 times, the internal water phase volume.
The above-described emulsifier and osmotic pressure regulator that may be added to the external water phase, and the subsequent procedures are the same as those described in paragraph (I) (i) above.
For producing microspheres by this method, a coacervating agent is added little by little to the organic solvent solution described in aqueous drying method paragraph (I) above, which contains a composition consisting of a biologically active substance or salt thereof, hydroxynaphthoic acid or salt thereof and biodegradable polymer or salt thereof, during stirring, to precipitate and solidify the microspheres. Said coacervating agent is added in an amount by volume of about 0.01 to 1,000 times, preferably about 0.05 to 500 times, and more preferably about 0.1 to 200 times, the oil phase volume.
Said coacervating agent may be any one, as long as it is a polymer, mineral oil or vegetable oil compound that is miscible in the organic solvent, and that does not dissolve the salt complex of the biologically active substance with the hydroxynaphthoic acid and biocompatible polymer. Specifically, useful coacervating agents include, for example, silicon oil, sesame oil, soybean oil, corn oil, cotton seed oil, coconut oil, linseed oil, mineral oil, n-hexane and n-heptane. These may be used in combination.
The microspheres thus obtained are collected, after which they are repeatedly washed with heptane etc. to remove the coacervating agent etc. other than the composition of the biologically active substance, hydroxynaphthoic acid and biodegradable polymer, followed by drying under reduced pressure. Alternatively, the microspheres are washed in the same manner as in aqueous drying method paragraph (I) (i) above, then freeze-dried and thermally dried.
For producing microspheres by this method, the organic solvent solution described in aqueous drying method paragraph (I) above, which contains a composition consisting of a biologically active substance or salt thereof, hydroxynaphthoic acid or salt thereof and biodegradable polymer or salt thereof, is sprayed via a nozzle into the drying chamber of a spray drier to volatilize the organic solvent in the fine droplets in a very short time, to yield microspheres. Said nozzle is exemplified by the double-fluid nozzle, pressure nozzle and rotary disc nozzle. The microspheres may be then freeze-dried and thermally dried as necessary after being washed in the same manner as that described in aqueous drying method paragraph (I) above.
For a dosage form other than the above-described microspheres, the organic solvent solution described in aqueous drying method paragraph (I) above, which contains a composition consisting of a biologically active substance or salt thereof, hydroxynaphthoic acid or salt thereof and biodegradable polymer or salt thereof, may be dried by evaporating the organic solvent and water, while the degree of vacuum is adjusted using a rotary evaporator or the like, followed by milling with a jet mill or the like, to yield microparticles.
The milled microparticles may be then freeze-dried and thermally dried after being washed in the same manner as that described in aqueous drying method paragraph (I) for microsphere production.
The microspheres or microparticles thus obtained enable drug release corresponding to the rate of decomposition of the biodegradable polymer or lactic acid-glycolic acid polymer used.
A biologically active substance or salt thereof is added to a solution of a hydroxynaphthoic acid or salt thereof in an organic solvent to a weight ratio falling within the above-described content range for biologically active substances, to yield an organic solvent solution of the hydroxynaphthoic acid of the biologically active substance.
Said organic solvent is the same as those described in paragraph (I) (i) above. When more than two kinds of organic solvents are used as a mixed solvent, the ratio of mixture is the same as those described in paragraph (I) (i) above.
Organic solvent removal for precipitation of a composition of a hydroxynaphthoic acid of the biologically active substance can be achieved by commonly known methods or methods based thereon. Such methods include, for example, the method in which the organic solvent is evaporated while the degree of vacuum is adjusted using a rotary evaporator or the like.
The thus-obtained composition of a hydroxynaphthoic acid of the biologically active substance can be again dissolved in an organic solvent to yield a sustained-release composition (microspheres or microparticles).
Said organic solvent is exemplified by halogenated hydrocarbons (e.g., dichloromethane, chloroform, dichloroethane, trichloroethane, carbon tetrachloride), ethers (e.g., ethyl ether, isopropyl ether), fatty acid esters (e.g., ethyl acetate, butyl acetate), and aromatic hydrocarbons (e.g., benzene, toluene, xylene). These solvents may be used in mixtures at appropriate ratios. Of these solvents, halogenated hydrocarbons are preferred, with greater preference given to dichloromethane.
The organic solvent solution containing the hydroxynaphthoic acid of the biologically active substance is then added to a water phase to form an O (oil phase)/W (water phase) emulsion, after which the solvent is evaporated from the oil phase to yield microspheres. For this operation, the water phase volume is normally chosen over the range from about 1 time to about 10,000 times, preferably from about 5 times to about 5,000 times, and more preferably from about 10 times to about 2,000 times, the oil phase volume.
An emulsifier, an osmotic pressure regulator and the following step is the same as those described in paragraph (I) (i).
Organic solvent removal can be achieved by commonly known methods or methods based thereon. Such methods include, for example, the method in which the organic solvent is evaporated under normal or gradually reduced pressure during stirring using a propeller stirrer, magnetic stirrer or the like, and the method in which the organic solvent is evaporated while the degree of vacuum is adjusted using a rotary evaporator or the like.
The thus-obtained microspheres are centrifuged or filtered to separate them, after which they are washed with distilled water several times to remove the free biologically active substance, hydroxynaphthoic acid, emulsifier etc. adhering to the microsphere surface, then again dispersed in distilled water etc. and freeze-dried.
To prevent mutual aggregation of particles during the production process, an anticoagulant may be added. Said anticoagulant is exemplified by water-soluble polysaccharides such as mannitol, lactose, glucose and starches (e.g., corn starch), amino acids such as glycine, and proteins such as fibrin and collagen. Of these substances, mannitol is preferred.
Where necessary, freeze-drying may be followed by heating under reduced pressure without causing mutual adhesion of microspheres, to further remove the water and organic solvent from the microspheres.
Although it varies depending on the amount of microspheres and other factors, heating time is normally about 12 hours to about 168 hours, preferably about 24 hours to about 120 hours, and more preferably about 48 hours to about 96 hours, after the microspheres reach a given temperature.
Any heating method can be used, as long as microsphere aggregates are uniformly heated.
Useful thermal drying methods include, for example, the method in which thermal drying is conducted in a constant-temperature chamber, fluidized bed chamber, mobile chamber or kiln, and the method using microwaves for thermal drying. Of these methods, the method in which thermal drying is conducted in a constant-temperature chamber is preferred. The microspheres obtained are relatively uniformly spherical and undergo little resistance during administration by injection so that needle clogging is unlikely. Also, possible use of thin injection needles mitigates patient pain at injection.
A biologically active substance or salt thereof is added to a solution of a hydroxynaphthoic acid or salt thereof in an organic solvent to a weight ratio falling within the above-described content range for biologically active substances, to yield an organic solvent solution of the hydroxynaphthoic acid of the biologically active substance, after which a sustained-release preparation (microspheres or microparticles) is prepared.
Said organic solvent is the same as those described in (I) (i). When more than two organic solvents are used as mixed solvents, the ratio of mixture is as same as those described in (I) (i).
The organic solvent solution containing the hydroxynaphthoic acid of the biologically active substance is then added to a water phase to form an O (oil phase)/W (water phase) emulsion, after which the solvent is evaporated from the oil phase to yield microspheres. For this operation, the water phase volume is normally chosen over the range from about 1 time to about 10,000 times, preferably from about 5 times to about 5,000 times, and more preferably from about 10 times to about 2,000 times, the oil phase volume.
The above-described emulsifier and osmotic pressure regulator that may be added to the external water phase, and the subsequent procedures are the same as those described in paragraph (IV) above.
The sustained-release composition of the present invention can be administered as such or in the form of various dosage forms prepared using it as a starting material, specifically as intramuscular, subcutaneous, visceral and other injectable preparations or implant preparations, nasal, rectal, uterine and other transdermal preparations, oral preparations [e.g., solid preparations such as capsules (e.g., hard capsules, soft capsules), granules and powders; liquids such as syrups, emulsions and suspensions] etc.
For example, the sustained-release composition of the present invention can be prepared as injectable preparations by suspending in water with a dispersing agent (e.g, surfactants such as Tween 80 and HCO-60, polysaccharides such as sodium hyaluronate, carboxymethyl cellulose and sodium alginate), a preservative (e.g., methyl paraben, propyl paraben), an isotonizing agent (e.g., sodium chloride, mannitol, sorbitol, glucose, proline) etc. to yield an aqueous suspension, or by dispersing in a vegetable oil such as sesame oil or corn oil to yield an oily suspension, whereby a practically useful sustained-release injectable preparation is obtained.
When the sustained-release composition of the present invention is used in the form of an injectable suspension, its particle diameter is chosen over such a range that the requirements concerning the degree of dispersion and needle passage are met. For example, the mean particle diameter normally ranges from about 0.1 to 360 xcexcm, preferably from about 0.5 to 150 xcexcm, and more preferably from about 1 to 100 xcexcm.
The sustained-release composition of the present invention can be prepared as a sterile preparation by such methods in which the entire production process is aseptic, the method using gamma rays for sterilization, and the method in which a preservative is added, which methods are not to be construed as limitative.
Because of low toxicity, the sustained-release composition of the present invention can be used as a safe pharmaceutical etc. in mammals (e.g., humans, bovines, swines, dogs, cats, mice, rats, rabbits).
Although varying widely depending on kind, content and dosage form of the active ingredient biologically active substance, and duration of release of the biologically active substance, target disease, subject animal species and other factors, the dose of the sustained-release composition may be set at any level, as long as the biologically active substance is effective. The dose of the active ingredient biologically active substance per administration can be preferably chosen as appropriate over the range from about 0.01 mg to 10 mg/kg body weight, more preferably from about 0.05 mg to 5 mg/kg body weight, per adult in the case of a 1-month release preparation.
The dose of the sustained-release composition per administration can be preferably chosen as appropriate over the range from about 0.05 mg to 50 mg/kg body weight, more preferably from about 0.1 mg to 30 mg/kg body weight per adult.
The frequency of administration can be chosen as appropriate, depending on kind, content and dosage form of the active ingredient biologically active substance, duration of release of the biologically active substance, target disease, subject animal species and other factors, e.g., once every several weeks, one every month or once every several months (e.g 3 months, 4 months, 6 months).
The sustained-release composition of the present invention is useful, depending on the biologically active substance which is contained in the sustained-release composition, as an agent for treating or preventing various kinds of diseases. When the biologically active substance is LH-RH derivatives, the sustained-release composition of the present invention is useful as an agent for treating or preventing of hormone-dependent diseases, especially sex hormone-dependent diseases, such as sex hormone-dependent cancer (e.g. prostatic cancer, hysterocarcinoma, breast cancer, hypophysoma, etc.), prostatic hypertrophy, endometriosis, hysteromyoma, precocious puberty, dysmenorrhea, amenorrhea, premenstrual syndrome, multilocular-ovary syndrome. The sustained-release composition of the present invention is also useful for an anticonceptive agent. When the rebound effect of after medication is used, the sustained-release composition of the present invention is useful as an agent for treating or preventing of infecundity. Further, the sustained-release composition of the present invention is useful as an agent for treating or preventing of sex hormone-nondependent, but LH-RH sensitive benign or cacoethic neoplasm.

Sustained release compositions, process for producing the same and utilization thereof


A squeeze spray device is described comprising a deformable bottle (1) for holding a liquid composition, a leave-in-place dispenser cap (5), and a dip-tube (50); said dispenser cap comprising:

The present invention relates to a squeeze-spray device and to a method of application of a liquid cosmetic composition to the human body.
Many forms of device for applying liquid cosmetic compositions are known. For spray application, propellant-driven aerosol devices are widely used. Alternatively, squeeze spray devices may be employed. Unfortunately, squeeze spray dispensing of liquid compositions commonly leads to poor spray quality, which, in turn, leads to unfavourable sensory perception when the composition is applied to the human body.
Squeeze spray devices of the prior art are generally unsophisticated devices and have the disadvantage of being prone to accidental discharge when inadvertently put under pressurexe2x80x94for example, when being carried in a handbag or a jacket pocket. Such accidental discharge not only wastes the product, but may also lead to inconvenient damage or cleaning requirements for the accidentally sprayed substrate.
Numerous publications exist in the literature on squeeze spray devices, some addressing one or more of the aforementioned problems. GB 2,284,202 (1994, Gene Stull) discloses a closure cap for a squeeze bottle, which, when opened, enables multi-stream spraying of the enclosed product. U.S. Pat. No. 5,240,149 (1993, Perfect-Valois Ventil) also discloses a closure cap for a squeeze bottle, although the outlet valve of disclosed closure cap may be opened simply by squeezing the bottle. U.S. Pat. No. 4,226,367 (1980, Hayes) discloses a squeeze spray device with a rotatable cap and alignable orifices. U.S. Pat. No. 4,102,476 (Ciba-Geigy) discloses a squeeze bottle device with an air check valve on the cover. U.S. Pat. No. 4,02,0,979 (1977, Summit Packaging Systems) discloses a squeeze spray device offering improved spray atomisation.
The present invention overcomes the aforementioned mentioned problems associated with many earlier squeeze spray devices. Other benefits may include ease of use, in particular one-handed use; robustness; and ease of manufacture.
When used to apply a liquid cosmetic composition to the human skin the devices described herein, in particular the preferred embodiments, lead to good sensory perception of the composition by the user.
According to a first aspect of the present invention, there is provided a squeeze spray device comprising a deformable bottle for holding a liquid composition, a leave-in-place dispenser cap attached to the deformable bottle, and a dip-tube, one end of said dip-tube being received by the dispenser cap and the other end terminating inside the deformable bottle; said dispenser cap comprising:
a) a base section, comprising a means for attachment to the deformable bottle, a segment defining at least one aperture and a receiver for the dip-tube on the interior face of said segment, the at least one aperture providing fluid connection between the interior of the deformable bottle and exterior to the base section, both via the dip-tube and otherwise; and
b) a moveable member, retained exterior to the base section, comprising a cover for said segment of the base section, said cover defining a spray hole;
the moveable member functioning to change the device between an inoperable state in which the spray hole is blocked from within by a blocking means that forms a part of the base section and an operable state in which the cover defining the spray hole is at a distance from said blocking means.
According to a second aspect of the present invention, there is provided a dispenser cap suitable for use as part of a squeeze spray device according to the first aspect of the invention.
According to a third aspect of the present invention, there is provided a method of topical application of a liquid cosmetic composition comprising the use of a squeeze spray device as described in the first aspect of the invention.
According to a fourth aspect of the present invention, there is provided a consumer product comprising a squeeze spray device as described in the first aspect of the invention containing a liquid composition.
With regard to the third and fourth aspects of the invention, the squeeze spray device of the present invention may be used with a variety of liquid compositions. It is suitable for topical application of a liquid cosmetic composition to the human body, in particular deodorant, antiperspirant, hair spray, and perfume compositions. Cosmetic compositions that are especially suitable are deodorant and antiperspirant compositions; the consumer may gain efficacy and/or sensory benefits with such compositions. The composition may be in the form of a homogeneous solution, an emulsion, or a suspension of one or more components in a liquid carrier material. Homogeneous solutions are the most preferred option because of their comparative stability and lack of problems with dispenser blockage.
The sensory and/or efficacy benefits referred to above are particularly noticeable when the composition used with the device is a liquid cosmetic composition for application to the human body that comprises a C2 to C4 alcohol, for example ethanol, propylene glycol, propanol, or iso-propanol. Suitable compositions typically comprise C2 to C4 alcohol at a level of from 5% to 95%, in particular from 25% to 80%, and especially from 40% to 75% by weight of the composition. Liquid compositions comprising ethanol are particularly suitable.
The deformable bottle may be of any shape; for example, it may be a straight-sided jar or flask, or it might have a waisted design. Preferably, the bottle is of tubular shape, with a base upon which it can stand upright, without falling over. The base may be essentially flat. It is preferred that the base is raised towards the interior of the bottle in a central region. The cross-section of the bottle is preferably circular. Another preferable feature is the presence of a neck portion, onto which the dispenser cap may be attached. Typical dimensions, excluding any neck portion, are from 5 cm to 20 cm for the height and, independently, from 3 cm to 8 cm for the diameter of a container of circular cross-section or from 3 cm to 12 cm for the greatest cross-sectional dimension of a container of non-circular cross-section. The sidewalls of the tubular container have a thickness enabling them to be deformable under hand-pressure, a typical thickness being from 0.3 mm to 1.2 mm. The base preferably has a thickness of from 0.6 mm to 2.5 mm.
The bottle may be made from any suitable plastic material; for example, polyethylene. A mixture of high- and low-density polyethylene may advantageously be employed.
When the deformable bottle is being used as part of a squeeze spray device suitable for topical application of a liquid cosmetic composition, its internal volume is typically from 10 cm3 to 300 cm3, particularly from 30 cm3 to 200 cm3, and especially from 50 cm3 to 150 cm3.
A dispenser cap is attached to the deformable bottle of the squeeze spray device. The dispenser cap is not removed during use of the squeeze spray devicexe2x80x94this is what is meant by xe2x80x9cleave-in-placexe2x80x9d.
The dispenser cap comprises a base section attached to the deformable bottle by some form of attachment means. A suitable attachment means is a screw-threaded tube designed to mate with a screw-threaded tube on a neck portion of the deformable bottle. Alternatively, the attachment means may be a friction fit onto the deformable bottle and/or lugs that interact with lugs on an adjoining surface on the base section. When the attachment means comprises screw-threaded elements as described above, it is preferred that there is also present a means of preventing over-tightening of the screw-threaded elements.
Preferably the base section comprises an outer skirt that fits over an upper part of the sidewall of the deformable bottle. This feature can aid the retention of the base section onto the deformable bottle. In particularly preferred embodiments, the lower edge of the outer skirt is shaped to fit into a correspondingly shaped indentation in the sidewall of the deformable bottle. This feature can prevent over-tightening of a screw-thread when the attachment means of the base section to the deformable bottle comprises such a feature. A suitable shape for the lower edge of the outer skirt is saddle-shaped.
The base section serves to largely close the deformable bottle; however, a segment of the base section defining at least one aperture provides fluid connection between the interior of the deformable bottle and exterior to said segment. The segment has a receiver for the dip-tube on its interior face (i.e., facing the interior of the deformable bottle) and the one or more apertures provide fluid connection with the interior of the deformable bottle both via the dip-tube and otherwise. The receiver is a structure that holds one end of the dip-tube in close proximity to the interior face of the base section without closing or blocking the bore of the dip-tube.
In certain embodiments, the receiver is in the interior of a tube projecting outwards from the base section (i.e., away from the deformable bottle). The outward-projecting tube generally has the segment of the base section defining at least one aperture at its outer end. Preferably, the interior of the tube comprises a portion of polygonal cross-section, triangular cross-section being particularly preferred. In these certain embodiments, the segment of the base section defining at least one orifice preferably defines a number of orifices equal to the number of sides of the portion of the interior of the tube of polygonal cross-section.
In preferred embodiments, the segment of the base section defining the at least one aperture is tubular in overall shape, preferably narrowing towards its outer end, ie. having a frusto-conical shape. The tubular segment may be located centrally at the outer end of an outward-projecting tube as described in the previous paragraph and share a common axis therewith. Preferably the tubular segment defining the at least one orifice is narrower than the outward-projecting tube, i.e. it has a smaller maximum outer dimension orthogonal to the axis than the corresponding outer dimension of the outward-projecting tube. Preferably, the at least one aperture is a slit running longitudinally through the sidewall of the tubular section.
The fluid connection between the interior of the deformable bottle and exterior to the segment of the base section defining the at least one aperture is provided both via the dip-tube and otherwise. The fluid connection other than via the dip-tube provides a means for air to pass between the interior of the deformable bottle and exterior to the segment of the base section defining the at least one aperture. When more than one aperture is defined by said segment of the base section, then two distinct embodiments are possible. In a first embodiment, each aperture independently provides fluid connection between the interior of the deformable bottle and exterior to said segment of the base section both via the dip-tube and otherwise. In a second embodiment, one or more apertures provide fluid connection between the interior of the deformable bottle and exterior to said segment of the base section without fluid connection via the dip-tube. In this latter embodiment, the fluid connection via the dip-tube is provided by one of the other apertures.
The number of orifices defined by the aforementioned segment of the base section is preferably at least three, three orifices being particularly preferred.
The base section comprises a blocking means for a spray hole (vide infra), said blocking means being a continuous surface on the exterior face of the base section of size sufficient to block said spray hole. Preferably the blocking means is at the outer end of a tubular element projecting outwards from the base section. More preferably, the tubular element comprises the segment of the base section that defines the at least one aperture. In the preferred embodiments in which said segment is tubular in overall shape, it is particularly preferred that said segment is closed at its outer end by the blocking means.
The dispenser cap also comprises a moveable member retained exterior to the deformable bottle. The moveable member is capable of changing the device from an inoperable to an operable state, and vice-versa. The member is retained in such a manner that it is not removed from the device, whether the device is in its operable state, its inoperable state, or in some intervening state. Preferably the moveable member is attached to the base section of the dispenser cap. The attachment means may comprise a tongue and groove arrangement and is preferably located at the side of the dispenser cap.
The moveable member comprises a cover for the segment of the base section defining at least one aperture, said cover defining a spray hole which is blocked from underneath by the aforementioned blocking means when the device is in its inoperable state. When the device is in its operable state, the cover defining the spray hole is at a distance from the blocking means, said distance being non-zero.
In preferred embodiments, the segment of the base section defining at least one aperture and the cover therefor form a swirl chamber where air and a liquid composition from the deformable bottle are mixed together prior to exiting through the spray hole. The air and the liquid composition are forced into the swirl chamber from the deformable body by squeezing thereof, in a preferred mode of operation.
In one embodiment, the device is changeable between its operable and its inoperable states by sliding the cover away from and towards the blocking means, respectively. In a second embodiment, the device is changeable between its operable and its inoperable states by screwing the cover away from and towards the blocking means, respectively. The former embodiment is preferred for ease of one-handed use.
In preferred embodiments, the cover defining the spray hole is a plate substantially orthogonal to the top-bottom axis of the device. The plate is moveable from a lower position in which the spray hole is blocked from underneath by the blocking means and the device is inoperable, to an upper position in which the spray hole is at a distance from the blocking means and the device is operable. The substantially orthogonal plate may be convex when viewed from the exterior and may be shaped to fit into an indentation in an outer portion of the base section.
The cover defining the spray may be attached to a side piece which is slid up or down the side of the dispensing cap to raise or lower the plate, thereby changing the device between its operable and inoperable state, respectively. A side piece of this nature is particularly advantageous for one-handed use.
In particularly preferred embodiments, the cover for the segment of the base section defining at least one aperture is a capped tube, the cap portion of which defines the spray hole and the interior side wall of which fits snugly over the exterior side wall of a tube projecting outwards from the base section, as previously described.
The moveable member may comprises a restraining means that interacts with the base section to prevent movement of the cover defining the spray hole beyond a pre-set distance from the blocking means, the device being in an operable state when the cover is at said pre-set distance from the blocking means. Said restraining means or another may also interact with the base section to provide resistance to the movement of the cover defining the spray hole away from the position in which the spray hole is blocked from within by the blocking means. Preferably the moveable member comprises a restraining means that can accomplish both of these functions.
The spray hole preferably has a maximum cross-sectional size of 1 mm or less, more preferably 0.6 mm or less. The spray hole preferably has a circular cross-sectional shape and preferably expands in cross-sectional size towards the exterior.
The dip-tube linking the dispensing cap with the lower region of the deformable plastic bottle is received by the dispensing cap in a way that enables flow of the liquid composition from the lower region of the bottle into the dispensing cap upon squeezing of the deformable bottle. The dip-tube preferably extends to the bottom of the deformable plastic bottle. Typically the dip-tube is manufactured from polyethylene and has a degree of flexibility. Typically the bore of the tube is from 0.5 mm to 1.5 mm and the thickness of the tube wall is from 0.2 mm to 0.7 mm.

Abstract: a) a base section (5B), comprising a means (15) for attachment to the deformable bottle, a segment (22) defining at least one aperture and a receiver (27B) for the dip-tube on the interior face of said segment (22), the at least one aperture providing fluid connection between the interior of the deformable bottle (1) and exterior to the base section (5B), both via the dip-tube and otherwise; and

Abstract: b) a moveable member (10), retained exterior to the base section, comprising a cover (14, 38) for said segment of the base section, said cover defining a spray hole (40);

Abstract: the moveable member (10) functioning to change the device between an inoperable state in which the spray hole (40) is blocked from within by a blocking means (26) that forms a part of the base section (5B) and an operable state in which the cover (14, 38) defining the spray hole (40) is at a distance from said blocking means (26).


Separate audio amplifier channels are provided with separate input signal level controls so that the output of one channel is unaffected by the input signal level of the other channel, and vice versa. A switch controls selection of the desired channel. Three channels also be provided and first and second switches control selection of any one of the three channels. As before, separate input signal level controls are provided for the three channels so that each is unaffected by the signal level control of either of the others. Two channels may be distortion channels, each with an input level control and also with an output level control. One channel may be a clean sound channel.

This invention relates to specialized amplifier systems for musical instrumentation, particularly for guitars, although its use is not limited thereto.
In performing contemporaneous popular music such as rock music, blues and the like, wherein the ensemble includes an electric lead instrument such as a guitar which at times may be playing a "background" or chordal line and, at other times, a solo or lead line, it has become customary for such instrument to be amplified differently for these two modes. In addition, it has been customary to provide for reverberation in either of these two modes. The solo line is normally referred to as the lead mode and the chordal line is normally referred to as the rhythm mode and for consistency, these terms will be used herein.
The rhythm mode refers to amplification which produces a relatively "clean" sound for the instrument in question, i.e., one which is free or substantially free of distortion; and the lead mode refers to amplification involving distortion of the sound of the instrument.
In the prior art, as exemplified for example by FIG. 1 of this application, the amplifier system comprises a series of cascaded amplifier stages for the rhythm mode and providing a "Volume I" potentiometer control at or near the input end of the cascaded stages and a "Master I" potentiometer control at the output of the final stage of the rhythm mode cascade. The amplifier system is switchable to the lead mode by switching in a further, distortion amplifier stage to the rhythm mode cascaded stages (while switching out the "Master I" control), such further stage having a "lead drive" potentiometer control at the input to the distortion stage and a "lead master" potentiometer control as the output of the distortion stage. As is conventional, the distortion stage comprises one or more high gain amplifiers operated non-linearly to provide generation of a composite signal in which half or more of the signal content may be distortion products (mostly even harmonics together with a substantial amount of odd harmonics).
Amplifier systems of this type have been very successful, but are sometimes difficult to use because the two modes are not independent. That is, the "Volume I" control sets the initial gain for both modes (or for further modes such as added reverberation or a preset attenuation of the lead mode) and this dependency makes it impossible always to obtain the desired output volumes for both a rhythm mode sound and for a lead mode sound. That is, for a very clean rhythm mode sound, the "Volume I" control must be set sufficiently low as to assure that the cascaded stages of the rhythm mode do not distort the audio signal input. This limits the maximum or full tilt distortion which may be produced in the lead mode. In consequence, some degree of compromise must be effected when setting the various interrelated volume controls. For example, assuming a very clean rhythm mode sound is to be achieved, this means that the "Volume I" control must be set to a relatively low value. If, now, a high degree of distortion is desired in the lead mode, it is limited by the setting of the "Volume I" control and the output volume of this limited distortion is likewise limited by the "Volume I" setting.
Consequently, this invention is directed to improvements in such amplifier systems.

Specialized amplifier systems for musical instruments