JP2002226365A - Method for producing pharmaceutical preparation including physiologically active substance - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing a pharmaceutical preparation that can release a constant amount of a physiologically active substance for a long period of time, as the initial excessive release of the physiologically active substance is inhibited immediately after the administration and is extremely reduced in deterioration of the active substance and extremely reduced in residue of an organic solvent. SOLUTION: In this method for producing the objective pharmaceutical preparation, a solid is formed from the physiologically active substance and a polymer and the resultant solid product is characteristically brought into contact with a high-pressure gas.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for producing a preparation containing a physiologically active substance, and more particularly to a method for producing a preparation containing a physiologically active substance and a polymer which are unstable to heat or a solvent.

[0002]

2. Description of the Related Art Peptidic or non-peptidic physiologically active substances are known to exhibit various pharmacological actions in living organisms, and are being applied to pharmaceuticals. However, these physiologically active substances generally have short half-lives in the living body, so that frequent administration is required, and the physical burden on the patient due to injection cannot be ignored. For example, growth hormone is a typical hormone that is originally produced and secreted by the anterior pituitary gland.In addition to working to promote body growth, it is involved in sugar and lipid metabolism, protein assimilation, cell proliferation and differentiation, etc.
It is a bioactive peptide with a wide variety of physiological actions.Currently, it is mass-produced by Escherichia coli using gene recombination technology and is widely used clinically as a drug worldwide.However, growth hormone has a short half-life in vivo. In order to maintain the effective blood concentration, frequent administration is necessary. Particularly in the case of pituitary dwarfism, long-term daily subcutaneous administration for infants or young patients for several months to 10 years or more is required. What is being done is the reality.

[0003] In order to address such problems inherent in physiologically active substances, various studies on drug delivery systems have been conducted. For example, it is a sustained release agent that releases a bioactive peptide over a long period of time. JP-A-8-217691
In the publication (WO96 / 07399), a water-soluble peptidic physiologically active substance is insoluble in water with an aqueous zinc chloride solution or the like.
A method for producing a sustained-release preparation containing a poorly water-soluble polyvalent metal salt and a biodegradable polymer is disclosed.

A sustained-release preparation using a biodegradable polymer suppresses the initial release of a physiologically active substance, in particular, the excessive release within one day, while maintaining the activity of the physiologically active substance, and also provides a long-term release. It is desirable to be able to arbitrarily control the release of a physiologically active substance over a wide range. On this issue,
Japanese Patent Application Laid-Open No. 11-322631 discloses that a bioactive peptide powder obtained by adding a water-miscible organic solvent and / or volatile salts to an aqueous solution of the bioactive peptide and freeze-drying the bioactive peptide is used as a biodegradable polymer. A method for producing a sustained-release preparation characterized by removing an organic solvent after dispersing in an organic solvent liquid is disclosed. Also, Japanese Patent Laid-Open No. 9-1
Japanese Patent No. 32524 discloses a method for producing a sustained-release microcapsule containing a physiologically active substance and a biodegradable polymer, wherein after microencapsulation, the biodegradable polymer has a glass transition temperature of about 24 to 120 hours or more. A method for producing a sustained-release preparation characterized by heating and drying is disclosed. These are methods for producing sustained-release preparations having extremely low residual organic solvents and excellent clinical properties as pharmaceuticals.

[0005]

However, the solvent removal method in the above production method requires a long time to remove the solvent, and therefore, there is room for improvement from the viewpoint of production cost in industrial practice. On the other hand, as a method for removing a solvent remaining in a substance (eg, a polymer) serving as a component when a pharmaceutical is formulated, a heat drying method, a vacuum drying method, and a flash drying method using a dry gas are known. However, such a method has a strong affinity for a solvent and may be insufficient in removing the solvent or decompose the substance for a substance unstable to heat. If the solvent to be removed has a high boiling point, the properties of the preparation may be impaired.

[0006]

Means for Solving the Problems The present inventors have intensively studied to solve the above-mentioned problems, and have found that a method for producing a sustained-release preparation containing a physiologically active substance and a biodegradable polymer, By contacting with a high-pressure gas for about 10 minutes to about 12 hours later, the initial release of an excessive amount of a physiologically active substance immediately after administration is dramatically suppressed, and a certain amount of physiologically active substance is maintained for a long period immediately after administration. It releases substances and has very little residual organic solvent.
They have found that a sustained-release preparation having very excellent properties can be produced, and based on these, the present invention has been completed.

That is, the present invention provides (1) a method for producing a preparation containing a physiologically active substance, which comprises forming a solid containing a physiologically active substance and a polymer, and contacting the solid with a high-pressure gas; ) The method according to the above (1), wherein the physiologically active substance is a physiologically active substance unstable to heat or a solvent, and (3) the physiologically active substance has a molecular weight of about 2,000 to about 50.
(4) the production method according to the above (1), wherein the physiologically active substance is a physiologically active peptide having a molecular weight of about 5,000 to about 500,000; (5) The method according to (4), wherein the bioactive peptide is human growth hormone, (6) the method according to (1), wherein the bioactive substance is a non-peptidic compound,
(7) The method according to the above (6), wherein the non-peptidic compound is a compound having an oxygen atom in the molecule, and (8) the above-mentioned (6), wherein the non-peptidic compound is a compound having an ether bond or a carbonyl group. Wherein the non-peptidic compound is of the formula (I)

Embedded image (Wherein, R ¹ represents a group capable of forming an anion or a group capable of changing to an anion; X represents that a phenylene group and a phenyl group are bonded directly or via a spacer having an atomic chain of 2 or less; represents an integer of 1 or 2, ring a denotes a benzene ring optionally further have a substituent, R ² represents a group capable of converting thereinto or a group capable of forming an anion, R ³
Represents a hydrocarbon residue which may be bonded via a hetero atom and may have a substituent) or a salt thereof, or (10). )
The process according to (6), wherein the non-peptidic compound is losartan, eprosartan, candesartan cilexetil, candesartan, valsartan, telmisartan, irbesartan, tasosartan or olmesartan,
(11) The production method according to the above (6), wherein the non-peptidic compound is candesartan, (12) the production method according to the above (1), wherein the polymer is biodegradable, and (13) the biodegradable polymer is α. The above (12), which is a homo- or copolymer of a hydroxycarboxylic acid or a mixture thereof;
(14) the biodegradable polymer is lactic acid /
(13) The production method according to the above (13), wherein the composition ratio of glycolic acid is about 100/0 to about 40/60 mol%, which is a lactic acid / glycolic acid homopolymer or copolymer, and (15) the biodegradable polymer is lactic acid. (17) the production method according to the above (12), wherein the weight average molecular weight of the biodegradable polymer is from about 3,000 to about 50,000; The process according to (1), wherein the solid is contacted with a high-pressure gas in a temperature range of about +20 to about -60 ° C based on the glass transition temperature of the polymer.
8) The solid is based on the glass transition temperature of the polymer.
(17) The method according to the above (17), wherein the solid is brought into contact with the high-pressure gas within a temperature range of about 0 to about -40 ° C. (20) The method according to (19), wherein the time for contacting the solid with the high-pressure gas is about 10 minutes to about 12 hours.
(21) The production method according to (1), wherein the high-pressure gas is an inert substance with respect to a physiologically active substance and a polymer, and (22) the production method according to (2), wherein the high-pressure gas is carbon dioxide.
(23) The method according to (1), wherein the pressure of the high-pressure gas is about 1 to about 7 MPa, and (24) the method (23), wherein the pressure of the high-pressure gas is about 1 to about 4 MPa. (25) The method according to (23), wherein the preparation is a sustained release microcapsule, and (26) the method according to (25), wherein the sustained release microcapsule is obtained by an underwater drying method. (27) a preparation obtained by the production method described in the above (1), (28) a sustained release microcapsule obtained by the production method described in the above (25), (29) a sustained release microcapsule described in the above (28) An injection comprising a capsule, (3
0) A method for suppressing initial release of a physiologically active substance, which comprises forming a solid containing a physiologically active substance and a polymer and contacting the solid with a high-pressure gas; and (31) containing a physiologically active substance and a polymer. It is intended to provide a method for suppressing deterioration of a physiologically active substance, which comprises forming a solid and bringing the solid into contact with a high-pressure gas.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION Examples of the physiologically active substance in the present invention include various drugs which have useful physiological activity for animals and plants and can be used as an agricultural or animal drug or clinically. As the physiologically active substance in the present invention, a physiologically active substance which is unstable to heat or a solvent is preferable. Here, a physiologically active substance which is unstable to heat or a solvent means decomposition, metabolism, inactivation or decomposition by heat or a solvent in a formulation step involving heating or contact with an organic solvent such as emulsification, desolvation or drying. Refers to a bioactive substance that denatures. As pesticides, for example, pest control agents, disease control agents,
Weed control agents, plant growth regulators, fertilizers and the like, and animal drugs include, for example, antibacterial agents, vitamins, hormones,
Examples include vaccines, marine preparations, insecticides and disinfectants, and pet drugs. It is important that the residual solvent is small as an ideal pesticide and animal medicine that is safe and environmentally friendly. Various drugs that can be used clinically are not particularly limited, for example, peptide compounds having a physiological activity, other antibiotics, antifungals, antihyperlipidemic drugs, antitumor drugs, antipyretics, analgesics, Anti-inflammatory, antitussive expectorant, sedative, muscle relaxant,
Antiepileptic, antiulcer, antidepressant, antiallergic,
Cardiotonic, arrhythmic, vasodilator, antihypertensive, diabetic, anticoagulant, hemostatic, antiplatelet, antituberculous,
Hormonal drugs, narcotic antagonists, bone resorption inhibitors, bone formation promoters, and angiogenesis inhibitors. Among them, peptidic or non-peptidic biologically active substances that produce analogs such as dimers and multimers or oxidized and deamidated forms by heat, and peptidic or non-peptidic substances that produce reactants with biodegradable polymers Non-peptide physiologically active substances are suitably used in the present invention.

[0009] Examples of the physiologically active peptide in the present invention include various peptides or proteins that have useful biological activity for mammals and can be used clinically. The "bioactive peptide" has a molecular weight of, for example, about 200 to 500,000 as a monomer, and preferably has a molecular weight of about 2,000 to 500,000.
000 are widely used. More preferably, a peptide having a molecular weight of 5,000 to about 500,000 is used. A typical example of the activity of a physiologically active peptide is a hormonal action. The bioactive peptide may be any of a natural product, a synthetic product, and a semi-synthetic product, and may be a derivative to an analog thereof. The mode of action of the physiologically active peptide may be either agonistic or antagonistic.
Examples of the physiologically active peptide in the present invention include peptide hormones, cytokines, peptidic neurotransmitters, hematopoietic factors, various growth factors, enzymes, polypeptide antibiotics, analgesic peptides, vaccines, and the like.

As peptide hormones, for example, insulin, somatostatin, somatostatin derivatives (Sandostatin, US Pat. Nos. 4,087,390 and 4,100)
093,574, 4,100,117 and 4,253,998), growth hormone (GH),
Natriuretic peptide, gastrin, prolactin,
Adrenocorticotropic hormone (ACTH), ACTH derivatives (eviratide etc.), melanocyte stimulating hormone (MS
H), thyroid hormone-releasing hormone (TRH), salts and derivatives thereof (see JP-A-50-112273 and JP-A-52-116465), thyroid-stimulating hormone (TSH), luteinizing hormone (LH), Follicle stimulating hormone (FSH), human chorionic gonadotropin (HC
G), thymosin (thymosin), motilin, vasopressin, vasopressin derivative ｛desmopressin [Journal of the Japan Endocrine Society, Vol. 54, No. 5, pp. 676-691 (1
978)]｝, oxytocin, calcitonin, parathyroid hormone (PTH), glucagon, secretin, pancreozymine, cholecystokinin, angiotensin, human placental lactogen, glucagon-like peptide (G
LP-1) and its derivatives (JP-A-6-80584).
JP-A-7-26995, EP658568, JP-A-8-245696, JP-A-8-269097, W
O97 / 15296, WO97 / 31943, WO
Nos. 98/19698, WO98 / 43658, JP-T10-511365, WO99 / 55310, JP-T11-513983, CA2270320, WO
No. 99/64061, JP-T-11-514772, JP-T-2000-500505, WO2000 / 6613
No. 8, WO2000 / 66142, WO2000 / 7
No. 8333, JP-A-2001-11095, Tissue
e Eng. 7 (1) 35-44 (2001), Diab
etologia 43 (10) 1319-1328 (200
0), WO2000 / 34331, WO2000 / 3
No. 4,332, U.S. Pat. No. 6,268,343, U.S. Publication No. 200101071, U.S. Publication No. 20010069.
No. 43, EP 0733644, WO2000 / 770
No. 39, WO99 / 43707, WO99 / 4334
No. 1, WO99 / 43706, WO99 / 43708
No., WO99 / 43705, WO99 / 29336
No., WO2000 / 37098, EP0969016
No. 5,981,488; U.S. Pat.
958,909, WO93 / 25579, WO98
No./43658, EP0869135, US Pat. No. 5,614,492, US Pat. No. 5,545,618.
No. 5,120,712; U.S. Pat.
118,666, WO95 / 05848, WO91
/ 11457, EP0708179, WO96 / 0
No. 6628, EP 0658568, WO 87/069
No. 41), glucose-dependent insulinotropic peptide (GIP), exendin and its derivatives (WO
2000/66629, WO2000 / 41546
No., WO99 / 07404, WO2000 / 0966
No. 6, U.S. Pat. No. 5,424,286), metastin and its derivatives (see WO2000 / 24890) and the like. Preferable peptide hormones include insulin and growth hormone.

The growth hormone (hereinafter referred to as GH) may be derived from any species, but is preferably human growth hormone (hereinafter referred to as hGH). In addition, natural sources extracted from the pituitary gland and the like are also used in the present invention, but preferably, recombinant GH (Japanese Patent Publication No.
No. 12996, Japanese Patent Publication No. 6-48987)
And more preferably a recombinant hGH having the same structure as a natural type having no methionine at the N-terminus. The GH may be a metal salt, but GH substantially containing no metal is also used. hGH not only has a molecular weight of about 22K daltons, but also has a molecular weight of about 20K daltons (JP-A-7-101877, JP-A-10-877).
265404). HG
H derivatives or their related proteins (WO99 / 0
3887).

As cytokines, for example, lymphokines, monokines and the like are used. Examples of lymphokines include interferons (alpha type, beta type, gamma type, etc.) and interleukins (eg, IL-
2,3,4,5,6,7,8,9,10,11,12
Etc.) are used. Monokines include, for example, interleukin-1 (IL-1), tumor necrosis factor (TN)
F) and the like are used. The cytokine is preferably lymphokine or the like, more preferably interferon or the like, particularly preferably interferon alpha or the like.

As the peptide neurotransmitter, for example, substance P, serotonin, GABA and the like are used. Examples of hematopoietic factors include erythropoietin (EP
O), colony stimulating factor (eg, G-CSF, GM-CS
F, M-CSF, etc.), thrombopoietin (TPO), platelet growth stimulating factor, megakaryocyte potentiator and the like. Examples of various growth factors include basic or acidic fibroblast growth factor (FGF) or a family of these (eg, EGF, TGF-α, TGF).
-Β, PDGF, acidic FGF, basic FGF, FGF-
9), nerve cell growth factor (NGF) or a family thereof (eg, BDNF, NT-3, NT-4, CN
TF, GDNF, etc.), insulin-like growth factors (eg, IG
F-1, IGF-2 etc.), factors involved in bone growth (BM
P) or their families are used.

As the enzyme, for example, superoxide dismutase (SOD), urokinase, tissue plasminogen activator (TPA), asparaginase, kallikrein and the like are used. As the polypeptide antibiotic, for example, polymyxin B, colistin, gramicidin, bacitracin and the like are used. As the analgesic peptide, for example, enkephalin, enkephalin derivatives (see U.S. Pat. No. 4,277,394, European Patent Application Publication No. 31567), endorphin, kyotorphin and the like are used. Examples of vaccines include influenza vaccine, Japanese encephalitis vaccine, rabies vaccine, hepatitis B vaccine, hepatitis A vaccine, cholera vaccine, DPT mixed vaccine,
Pneumococcal vaccine, diphtheria vaccine, tetanus vaccine, polio vaccine, prostate-specific antigen vaccine and the like are used. Other bioactive peptides include thymopoietin, dynorphin, bombesin, caerulein, thymostimulin, thymic humoral factor (THF), blood thymic factor (FTS) and derivatives thereof (US Pat. No. 4,22
9,438), and other thymic factors [Ayumi of Medicine, 125, 10, 835-843 (1
983)], peptides having an antagonistic action on neurotensin, bradykinin and endothelin (European Patent Publication Nos. 436189, 457195, 496452, JP-A-3-94692, 3-13).
No. 0299).

In the present invention, when the bioactive peptide contains a metal, if necessary, the metal contained in the bioactive peptide may be removed in advance.
As a method for removing the metal, a known method is used.
For example, an aqueous hydrochloric acid solution of insulin is dialyzed against water or an ammonium acetate solution and then lyophilized to obtain insulin in an amorphous state and a minimum amount of metal.

The non-peptide physiologically active substance in the present invention includes, for example, nourishing tonic health drugs, antipyretic analgesic and anti-inflammatory drugs,
Psychotropics, anxiolytics, antidepressants, hypnotics, antispasmodics, central nervous system drugs, brain metabolism improvers, antiepileptics, sympathomimetics, gastrointestinal drugs, antacids, antiulcers, antitussives Expectorants, antiemetics, respiratory stimulants, bronchodilators, antiallergic drugs, oral drugs, antihistamines, inotropic agents, antiarrhythmic drugs, diuretics, antihypertensive drugs, vasoconstrictors, coronary vasodilators,
Peripheral vasodilators, antihyperlipidemic agents, bile agents, antibiotics, chemotherapeutic agents, antidiabetic agents, osteoporosis agents, skeletal muscle relaxants, analgesics, hormonal agents, alkaloid drugs, sulfa drugs , Gout remedies, anticoagulants, antineoplastics,
One or two selected from drugs for treating Alzheimer's disease
More than one component may be included.

Examples of the nutritional tonic health medicine include vitamin A, vitamin D, vitamin E (such as d-α-tocopherol acetate), vitamin B ₁ (such as dibenzoylthiamine and fursultiamine hydrochloride), and vitamin B ₂ (butyric acid). amino acid; minerals such as calcium, magnesium, iron; riboflavin, etc.), vitamin B ₆ (pyridoxine hydrochloride), vitamin C (ascorbic acid, sodium L- ascorbate), vitamin vitamin B ₁₂ (such as acetic hydroxocobalamin) Oligosaccharides, crude drugs and the like can be mentioned. Antipyretic analgesics and anti-inflammatory drugs include, for example, aspirin, acetaminophen, ethenzamide, ibuprofen, diphenhydramine hydrochloride, dl-chlorpheniramine maleate, dihydrocodeine phosphate, noscapine,
Methyl ephedrine hydrochloride, phenylpropanolamine hydrochloride, caffeine, anhydrous caffeine, tolfenamic acid,
Examples include mefenamic acid, diclofenac sodium, flufenamic acid, salicylamide, aminopyrine, ketoprofen, indomethacin, bucolome, pentazocine and the like. Psychotropic drugs include, for example, chlorpromazine, reserpine and the like. Examples of the anxiolytic include alprazolam, chlordiazepoxide, diazepam and the like. Examples of the antidepressant include imipramine, maprotiline, amphetamine and the like.

Hypnotic sedatives include, for example, estazolam, nitrazepam, diazepam, perlapine, phenobarbital sodium and the like. Examples of antispasmodics include scopolamine hydrobromide, diphenhydramine hydrochloride, papaverine hydrochloride and the like. Examples of central nervous system drugs include citicoline, rothyrenine and the like. Examples of the cerebral metabolism improving agent include vinpocetine, meclofenoxate hydrochloride and the like. Antiepileptic agents include, for example, phenytoin, carbamazepine and the like. Sympathetic stimulants include, for example, isoproterenol hydrochloride and the like. Examples of the gastrointestinal drug include a stomach digestive agent such as gentian, cassava, homica, oak, spruce, conzulango, and calyx oil; Examples of the antacid include magnesium carbonate, sodium hydrogencarbonate, magnesium aluminate metasilicate, synthetic hydrotalcite, precipitated calcium carbonate, magnesium oxide and the like. Examples of the anti-ulcer agent include lansoprazole, omeprazole, rabeprazole, pantoprazole, famotidine, cimetidine, ranitidine hydrochloride and the like.

Examples of antitussive expectorants include cloperastine hydrochloride, dextromeltophan hydrobromide, theophylline, potassium guaiacol sulfonate, guaifenesin, codeine phosphate and the like. Examples of the antiemetic include difenidol hydrochloride, metoclopramide and the like. Examples of the respiratory enhancer include levallorphan tartrate. As a bronchodilator,
Examples include theophylline and salbutanol sulfate. Examples of antiallergic agents include amlexanox, seratrodast, and the like. Examples of the oral medicine include oxytetracycline, triamcinolone acetonide, chlorhexidine hydrochloride, lidocaine and the like. Examples of the antihistamine include diphenhydramine hydrochloride, promethazine, isotipendyl hydrochloride, dl-chlorpheniramine maleate, and the like. Examples of cardiotonic agents include caffeine, digoxin and the like. Examples of the antiarrhythmic agent include procainamide hydrochloride, propranolol hydrochloride, pindolol and the like. Diuretics include, for example, isosorbide, furosemide and the like. Examples of antihypertensive agents include delapril hydrochloride, captopril, hexamethonium bromide, hydralazine hydrochloride, labetalol hydrochloride, manidipine hydrochloride, losartan, eprosartan, candesartan cilexetil (TCV-116), candesartan (CV
-11974), valsartan, telmisartan, irbesartan, tasosartan, olmesartan and the like. As a hypotensive agent, preferably candesartan,
Candesartan cilexetil and the like, particularly preferably candesartan and the like.

Examples of the vasoconstrictor include phenylephrine hydrochloride. Examples of the coronary vasodilator include carbochromene hydrochloride, molsidomine, verapamil hydrochloride and the like. Peripheral vasodilators include, for example, cinnarizine and the like. As antihyperlipidemic agents,
For example, cerivastatin sodium, simvastatin,
Pravastatin sodium and the like. Bile agents include, for example, dehydrocholic acid, trepibutone and the like. Examples of the antibiotics include cephalexin, amoxicillin, pivmecillinam hydrochloride, cefotiam hydrochloride, cefozopran hydrochloride, cefmenoxime hydrochloride, cefusrodin sodium and other cephem antibiotics; ampicillin, cyclacillin, sulbenicillin sodium,
Synthetic antibacterial agents such as nalidixic acid and enoxacin; monobactam antibiotics such as carmonam sodium; penem antibiotics and carbapenem antibiotics. Examples of the chemotherapeutic agent include sulfamethizole hydrochloride, thiazosulfone and the like. Examples of antidiabetic agents include tolbutamide, voglibose, pioglitazone (hydrochloride), troglitazone, 5-[[4- [2-
(Methyl-2-pyridinylamino) ethoxy] phenyl] methyl] -2,4-thiazolidinedione (BRL
-49653), acarbose, miglitol, emiglitate and the like. Examples of the agent for osteoporosis include ipriflavone and the like. Skeletal muscle relaxants include, for example, methocarbamol. Examples of the antitussive include meclizine hydrochloride, dimenhydrinate and the like.

Examples of hormonal agents include liothyronine sodium, dexamethasone sodium phosphate, prednisolone, oxendrone, leuprorelin acetate and the like. Examples of the alkaloid narcotics include opium, morphine hydrochloride, tocon, oxycodone hydrochloride, opium alkaloid hydrochloride, cocaine hydrochloride and the like. Examples of the sulfa drug include sulfamine, sulfamethizole and the like. Examples of gout remedies include allopurinol, colchicine and the like. Examples of the anticoagulant include dicoumarol and the like. Examples of the antineoplastic agent include 5-fluorouracil, uracil, and mitomycin. Examples of the drug for treating Alzheimer's disease include idebenone, vinpocetine and the like.

The non-peptidic physiologically active substance in the present invention is preferably a compound having an oxygen atom in the molecule, specifically, a compound having an ether bond or a carbonyl group. Such compounds include those of formula (I)

Embedded image Or a salt thereof. In the above formula (I), as a group capable of forming an anion as R ¹ (a group having a hydrogen atom that can be released as a proton),
For example, (1) carboxyl group, (2) tetrazolyl group, (3) trifluoromethanesulfonic acid amide group (-
NHSO ₂ CF ₃ ), (4) phosphoric acid group, (5) sulfonic acid group, (6) 5 to 7 members (preferably 5 to 6 members) containing one or more of N, S, and O )) A monocyclic optionally substituted heterocyclic residue.

The above-mentioned "5- to 7-membered (preferably 5- to 6-membered) monocyclic optionally substituted heterocyclic residue containing one or more of N, S and O" Is, for example,

Embedded image And the bond between the heterocyclic residue represented by R ¹ and the phenyl group to which the heterocyclic residue is bonded, as described above when g in the above formula represents —NH— or the like. Carbon-
The bond may be formed not only through a carbon bond but also through one of a plurality of nitrogen atoms. For example, if R ¹ is

Embedded image

Embedded image In the above formula, g is _{-CH 2 -, - NH -,} - O- or -
S (O) _m −,> = Z,> = Z ′ and> =
Z ″ is a carbonyl group, a thiocarbonyl group or a sulfur atom which may be oxidized (eg, S, S (O),
S (O) ₂ or the like (preferably a carbonyl or thiocarbonyl group, more preferably a carbonyl group),
m represents 0, 1 or 2.

The heterocyclic residue represented by R ¹ includes, for example,-as a proton donor such as an oxadiazolone ring, an oxadiazolothione ring or a thiadiazolone ring.
A group obtained by removing one hydrogen atom from a ring having both an NH- or -OH group and a carbonyl group, a thiocarbonyl group, a sulfinyl group, or the like as a proton acceptor is preferable. Further, the heterocyclic residue represented by R ^1, may form a fused ring group bonded to a cyclic substituent. The preferred heterocyclic residue represented by R ^1, 5 or 6-membered Rings and 5-membered ring residues are preferred. The heterocyclic residue represented by R ¹ includes a group represented by the formula:

Embedded image [Wherein, i represents -O- or -S-, j is> = O,
> = S or> = S (O) _m , wherein m is as defined above] (among others, 2,5-dihydro-
5-oxo-1,2,4-oxadiazol-3-yl, 2,5-dihydro-5-thioxo-1,2,4-oxadiazol-3-yl, 2,5-dihydro-5 Oxo-1,2,4-thiadiazol-3-yl, especially 2,5-dihydro-5-oxo-1,2,4-oxadiazol-3-yl) and the like are preferable. The heterocyclic residue (R ¹ ) has a tautomer as shown below. For example,

Embedded image There are three tautomers of a ', b' and c 'such as

Embedded image The heterocyclic residue represented by the above includes all of the above a ′, b ′ and c ′.

The group capable of forming an anion as R ¹ is
At a substitutable position, an optionally substituted lower (C _1-4 ) alkyl group or an acyl group (eg, lower (C
_2-5 ) alkanoyl, benzoyl, etc.). Examples of the optionally substituted lower (C _1-4 ) alkyl group include (1) a halogen atom, nitro, lower (C _1-4 ) alkyl, lower (C _1-4 ) alkoxy and the like. Lower (C) which may be substituted with 1 to 3 phenyl groups
_1-4 ) alkyl group (eg, methyl, triphenylmethyl, p-methoxybenzyl, p-nitrobenzyl, etc.), (2) lower (C _1-4 ) alkoxy-lower (C
_1-4 ) an alkyl group (eg, methoxymethyl, ethoxymethyl, etc.), (3) a formula —CH (R ⁴ ) —OCOR ⁵ [wherein R ⁴ is (a) hydrogen, and (b) 1-6 carbon atoms. Linear or branched lower alkyl groups (eg, methyl, ethyl, n
-Propyl, isopropyl, n-butyl, isobutyl, t
-Butyl, n-pentyl, isopentyl, neopentyl, etc.), (c) a linear or branched lower alkenyl group having 2 to 6 carbon atoms or (d) a cycloalkyl group having 3 to 8 carbon atoms (eg, cyclopentyl, cyclohexyl, shows cycloheptyl, etc.), R ⁵ is (a) a linear or branched lower alkyl group (examples of the carbon atoms 1-6, methyl, ethyl, n- propyl, isopropyl, n- butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, etc.), (b) a linear or branched lower alkenyl group having 2-6 carbon atoms, (c) a 3-carbon atom
8 cycloalkyl group (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.) or an optionally substituted aryl group (eg, halogen atom, nitro, lower (C _1-4 ) alkyl, lower (C _1-4 ) alkoxy Etc., which may have a phenyl or naphthyl group)
A lower alkyl group having 1 to 3 carbon atoms (eg, benzyl, p-chlorobenzyl, phenethyl, cyclopentylmethyl, cyclohexylmethyl, etc.), (d) cycloalkyl having 3 to 8 carbon atoms, or Good aryl groups (eg, halogen atom, nitro, lower (C
_1-4 ) a lower alkenyl group having 2-3 carbon atoms (eg, vinyl such as cinnamyl, propenyl) substituted by an alkyl, a phenyl or naphthyl group which may have a lower (C _1-4 ) alkoxy or the like; , Allyl, those having an alkenyl moiety such as isopropenyl), (e) an optionally substituted aryl group (eg, phenyl, p-tolyl,
Halogen atom such as naphthyl, nitro, lower (C _1-4 )
Alkyl, a phenyl or naphthyl group optionally having a lower (C _1-4 ) alkoxy, etc.), (f) a linear or branched lower alkoxy group having 1-6 carbon atoms (eg, methoxy, ethoxy) , N-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy,
t-butoxy, n-pentyloxy, isopentyloxy, neopentyloxy, etc.), (g) a linear or branched lower alkenyloxy group having 2-8 carbon atoms (eg, allyloxy, isobutenyloxy, etc.), (h) 3-8 carbon atoms
(E.g., cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, etc.), (i) cycloalkyl having 3-8 carbon atoms (e.g., cyclopentyl, cyclohexyl, cycloheptyl, etc.) or optionally substituted aryl Group (eg, halogen atom, nitro, lower (C _1-4 ) alkyl, lower (C
_1-4 ) a lower alkoxy group having 1 to 3 carbon atoms (eg, methoxy such as benzyloxy, phenethyloxy, cyclopentylmethoxy, cyclohexylmethoxy, ethoxy, etc.) substituted with a phenyl or naphthyl group optionally having an alkoxy or the like n-propoxy, isopropoxy, etc. having an alkoxy moiety), (j) 3 carbon atoms
-8 cycloalkyl (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.) or an optionally substituted aryl group (eg, halogen atom, nitro, lower (C _1-4 ) alkyl, lower (C _1-4 ) alkoxy Etc., which may have a phenyl or naphthyl group)
And a lower alkenyloxy group having 2 to 3 carbon atoms (eg, a alkenyloxy group such as vinyloxy, propenyloxy, allyloxy, isopropenyloxy, etc. having cinnamyloxy) or (k) an aryloxy group which may be substituted (E.g., phenoxy, p-nitrophenoxy, naphthoxy and other halogen atoms, nitro, lower (C _1-4 ) alkyl, lower (C _1-4 ) alkoxy, etc., and optionally phenoxy or naphthoxy groups). And the like). Also, R
^The group capable of forming an anion as ¹ is protected by the above-mentioned optionally substituted lower (C _1-4 ) alkyl group or acyl group (eg, lower (C _2-5 ) alkanoyl, benzoyl, etc.). Other than a group, at a substitutable position,
Also optionally substituted lower (C _1-4) alkyl group (has been "optionally substituted exemplified as a protective group for the group capable of forming an anion as R ¹ mentioned above a lower (C _1-4) The same as the “alkyl group”), a halogen atom, nitro, cyano, lower (C _1-4 ) alkoxy, amino optionally substituted with one or two lower (C _1-4 ) alkyl, and the like. May have a substituent. In the above formula, a group capable of forming an anion as R ¹ (a group having a hydrogen atom which can be released as a proton) is converted under a biological or physiological condition (for example, oxidation by an in vivo enzyme or the like). ,
The group may be a group capable of forming an anion by an in vivo reaction such as reduction or hydrolysis (a so-called prodrug), or a cyano or N-hydroxycarbamimidoyl group (-C ( _{= N-OH) -NH 2)} ,
Alternatively, (1) a carboxyl group, (2) a tetrazolyl group, and (3) a trifluoromethanesulfonic acid amide group (—NHSO ₂ CF) protected with an optionally substituted lower (C _1-4 ) alkyl group or an acyl group, respectively. ₃ ),
(4) phosphate group, (5) sulfonic group, (6) N, S,
Like a 5- to 7-membered (preferably 5- to 6-membered) monocyclic optionally substituted heterocyclic residue containing one or more of O, R ¹ (So-called synthetic intermediate). R ¹ is optionally substituted lower (C _1-4 ) alkyl (eg, methyl, triphenylmethyl, methoxymethyl, ethoxymethyl, p-methoxybenzyl, p-nitrobenzyl, etc.) or an acyl group (eg, Carboxyl, tetrazolyl or 2,5-dihydro-5-oxo-1, which may be protected with lower (C _2-5 ) alkanoyl, benzoyl and the like.
2,4-oxadiazol-3-yl (preferably tetrazolyl) or cyano or N-hydroxycarbamimidoyl (preferably cyano) is preferable, and cyano is particularly preferably used.

In the above formula, X represents that an adjacent phenylene group and a phenyl group are bonded directly or via a spacer having an atomic chain of 2 or less (preferably a direct bond). Any of divalent chains having 1 or 2 atoms constituting the straight chain portion may be used, and may have a side chain. Specifically, lower (C _1-4 ) alkylene having 1 or 2 atoms constituting a straight chain portion, —CO—, —O—, —S—, —NH—, —C
_{O-NH -, - O-} CH 2 -, - S-CH 2 -, - CH
= CH- and the like. In the above formula, n is 1 or 2
(Preferably 1). In the above formula, ring A represents a benzene ring which may further have a substituent other than a substituent R ^2, examples of the substituent, for example, (1) halogen (e.g., F, Cl, Br, etc.) , (2) cyano, (3) nitro, (4) optionally substituted lower (C _1-4 ) alkyl, (5) lower (C _1-4 ) alkoxy, (6) optionally substituted Amino group (eg, amino, N-lower (C
_1-4 ) alkylamino (eg, methylamino, etc.),
N, N-di-lower (C _1-4 ) alkylamino (eg, dimethylamino), N-arylamino (eg, phenylamino), alicyclic amino (eg, morpholino, piberidino, piperazino, N-phenyl) Piperazino etc.)), (7) Formula -CO-D 'wherein D' is a hydroxyl group or an alkyl moiety is a hydroxyl group, lower (C _1-4 ) alkoxy, lower (C _2-6 ) alkanoyloxy ( For example, acetoxy, pivaloyloxy, etc.), lower (C _1-6 ) alkoxycarbonyloxy (eg, methoxycarbonyloxy, ethoxycarbonyloxy, etc.) or lower (C
_3-6 ) lower (C _1-4 ) alkoxy which may be substituted by cycloalkoxycarbonyloxy (eg, cyclohexyloxycarbonyloxy, etc.), or (8) may be substituted Low (C
_1-4 ) alkyl (the same as the above-mentioned "optionally substituted lower (C _1-4 ) alkyl group" exemplified as the protecting group for the group capable of forming an anion as R ¹ ) ) Or acyl (eg, lower ( _C2-5 ) alkanoyl, benzoyl, etc.) optionally protected tetrazolyl, trifluoromethanesulfonic acid amide group,
Examples include a phosphoric acid group or a sulfonic acid group. These substituents are located at substitutable positions on the benzene ring,
In addition to the substituent R ² , the substituent which the ring A further has may be a lower (C _1-4 ) alkyl which may be substituted (eg, a hydroxyl group, a carboxyl group, a halogen, etc.) Optionally substituted by lower (C
_1-4) alkyl, etc.), are preferred, such as halogen, ring A other than the substituent R ² is more preferably has no substituent.

In the above formula, R²Forming anions as
Group (having a hydrogen atom that can be released as a proton)
Groups) include, for example, (1) esterification or amide
Carboxyl group which may be converted to (2) tetrazoli
(3) trifluoromethanesulfonic acid amide group
(-NHSO₂CF₃), (4) phosphate group, (5) sulf
And sulfonic acid groups.
A lower alkyl group (R as described above)¹As an anion
"Substituted as exemplified as a protecting group for a group capable of forming
Low-grade (C_1-4) Alkyl group "
Or an acyl group (eg, lower)
(C_2-5) Protected by alkanoyl, benzoyl, etc.)
Under biological or physiological conditions (eg,
For example, oxidation, reduction or hydrolysis by enzymes in the body
In vivo reactions) or chemically anions
Any group that can be formed or can be changed to
Is also good. R²Esterified or amidated as
Examples of the optionally available carboxyl include, for example, a compound of the formula -CO-D
[Wherein D is (1) a hydroxyl group, and (2) it may be substituted.
Amino (for example, amino, N-lower (C_1-4) Al
Killamino, lower N, N-di (C_1-4) Alkylam
Or (3) optionally substituted alkoxy
{Eg, (i) Even if the alkyl moiety is hydroxyl or substituted
Good amino (eg, amino, N-lower (C_1-4) Archi
Ruamino, N, N-di-lower (C_1-4) Alkylam
, Piperidino, morpholino, etc.), halogen, lower
(C_1-6) Alkoxy, lower (C_1-6) Alkyl
Oh, low grade (C_3-8) Cycloalkoxy or substituted
Dioxolenyl (e.g., 5-methyl-2-
Oxo-1,3-dioxolen-4-yl)
Low-grade (C_1-6) Alkoxy groups, also
Is (ii) the formula -O-CH (R⁶) -OCOR⁷(In the formula,
R ⁶Is (a) hydrogen, (b) a straight chain having 1 to 6 carbon atoms or
A branched lower alkyl group (eg, methyl, ethyl, n-propyl)
Propyl, isopropyl, n-butyl, isobutyl, t-butyl
Tyl, n-pentyl, isopentyl, neopentyl
And (c) a linear or branched lower chain having 2 to 6 carbon atoms.
Alkenyl group or (d) cycloalkyl having 3 to 8 carbon atoms
Group (eg, cyclopentyl, cyclohexyl, cyclohexyl)
Etc.), R⁷Is (a) a linear chain having 1-6 carbon atoms
Or a branched lower alkyl group (eg, methyl, ethyl
, N-propyl, isopropyl, n-butyl, isobutyl
, Sec-butyl, t-butyl, n-pentyl, isopene
(B) a straight chain having 2 to 6 carbon atoms.
Or a branched lower alkenyl group, (c)
8 cycloalkyl groups (eg, cyclopentyl, cyclohexyl)
Xyl, cycloheptyl, etc.) or substituted
Aryl groups (eg, halogen atom, nitro, lower
(C_1-4) Alkyl, lower (C_1-4) Alkoxy
A phenyl or naphthyl group which may have any number)
A lower alkyl group having 1 to 3 carbon atoms substituted with
Jill, p-chlorobenzyl, phenethyl, cyclopent
Methyl, cyclohexylmethyl, etc.), (d) carbon number
3-8 cycloalkyl or optionally substituted
Aryl group (eg, halogen atom, nitro, lower (C
_1-4) Alkyl, lower (C_1-4) Alkoxy etc.
Phenyl or naphthyl group which may be present)
Substituted lower alkenyl group having 2-3 carbon atoms (eg, cinna
Vinyl such as mill, propenyl, allyl, isopropenyl
Such as those having an alkenyl moiety such as
Optionally substituted aryl groups (eg, phenyl, p-tolyl,
Halogen atoms such as naphthyl, nitro, lower (C_1-4)
Alkyl, lower (C_1-4) Having alkoxy etc.
Phenyl or naphthyl group, etc.), (f) carbon
A linear or branched lower alkoxy group of the formula 1-6
(E.g., methoxy, ethoxy, n-propoxy, isopro
Poxy, n-butoxy, isobutoxy, sec-butoxy,
t-butoxy, n-pentyloxy, isopentyloxy
, Neopentyloxy, etc.), (g) having 2-8 carbon atoms
A linear or branched lower alkenyloxy group (eg,
Roxy, isobutenyloxy, etc.), (h) 3-8 carbon atoms
A cycloalkyloxy group (eg, cyclopentyloxy)
, Cyclohexyloxy, cycloheptyloxy
), (I) cycloalkyl having 3 to 8 carbon atoms (eg, cycloalkyl)
Lopentyl, cyclohexyl, cycloheptyl, etc.)
Or an optionally substituted aryl group (eg, halogen
Atom, nitro, lower (C_1-4) Alkyl, lower (C
_1-4) Phenyl which may have alkoxy or the like
Or lower having 1 to 3 carbon atoms, which is substituted with
Alkoxy groups (eg, benzyloxy, phenethyloxy,
Clopentyl methoxy, cyclohexyl methoxy, etc.
Methoxy, ethoxy, n-propoxy, isopropoxy
Such as those having an alkoxy moiety), (j) having 3 carbon atoms
-8 cycloalkyl (eg, cyclopentyl, cyclohexyl)
Xyl, cycloheptyl, etc.) or substituted
Aryl groups (eg, halogen atom, nitro, lower
(C_1-4) Alkyl, lower (C_1-4) Alkoxy
A phenyl or naphthyl group which may have any number)
C 2-3 lower alkenyloxy group substituted by
(Eg, vinyloxy such as cinnamiroxy, propeniroki
Alkenyls such as si, allyloxy, and isopropenyloxy
Or (k) even if it is substituted
Good aryloxy groups (eg, phenoxy, p-nitroph
Halogen atom such as phenoxy, naphthoxy, nitro, lower
(C_1-4) Alkyl, lower (C_1-4) Alkoxy
Phenoxy or naphthoxy group which may have
Etc.) or a group represented by｝)
And the like. R²Are esterified
Carboxyl which may be preferred, as specific examples thereof
Is, for example, -COOH and a salt thereof, -COOMe,-
COOEt, -COOTBu, -COOPr, Pivaloyl
Xymethoxycarbonyl, 1- (cyclohexyloxy
Carbonyloxy) ethoxycarbonyl, 5-methyl-
2-oxo-1,3-dioxolen-4-ylmethoxy
Carbonyl, acetoxymethoxycarbonyl, propio
Niloxymethoxycarbonyl, n-butylyloxymethoxy
Cicarbonyl, isobutylyloxymethoxycarbonyl,
1- (ethoxycarbonyloxy) ethoxycarbonyl,
1- (acetoxy) ethoxycarbonyl, 1- (isobu
Tyryloxy) ethoxycarbonyl, cyclohexylcar
Bonyloxymethoxycarbonyl, benzoyloxime
Toxicylcarbonyl, cinnamyloxycarbonyl, cyclo
And pentylcarbonyloxymethoxycarbonyl.
Under biological or physiological conditions (eg, in vivo
Biological inversion such as oxidation / reduction or hydrolysis by enzymes
Anion) or chemically anions (eg, COO⁻,
Or a derivative thereof)
Any group can be used as long as it is a carboxyl group or
Or a prodrug thereof. R above²age
In the formula, -CO-D [where D is (1) a hydroxyl group or
(2) Alkyl moiety is hydroxyl group, amino, halogen, lower
(C_2-6) Alkanoyloxy (eg, acetooxy,
Pivaloyloxy), lower (C_3-8) Cycloal
Canoyloxy, lower (C_1-6) Alkoxycarboni
Luoxy (eg, methoxycarbonyloxy, ethoxyca
Rubonyloxy, etc.), lower (C_3-8) Cycloalco
Xycarboniloxy (eg, cyclohexyloxycarbo
Low-grade (C_1-4) Alkoxy or
Low grade (C_3-8) Even if substituted with cycloalkoxy
Good low grade (C_1-4) Represents alkoxy]
Groups are preferred, and lower (C)_1-4) Alkyl (good
Preferably methyl or ethyl)
Ruboxyl is preferred.

In the above formula, the “hydrocarbon residue” in the “hydrocarbon residue which may be bonded via a hetero atom and has a substituent” represented by R ³ includes, for example,
(1) alkyl group, (2) alkenyl group, (3) alkynyl group, (4) cycloalkyl group, (5) aryl group,
(6) An aralkyl group and the like are mentioned, and among them, an alkyl group, an alkenyl group and a cycloalkyl group are preferable. The alkyl group of the above (1) is a lower alkyl group having about 1 to 8 carbon atoms, which may be linear or branched, for example, methyl, ethyl, propyl, isopropyl,
Butyl, isobutyl, sec-butyl, t-butyl, pentyl, i-pentyl, hexyl, heptyl, octyl and the like. As the alkenyl group of the above (2),
The lower alkenyl group having about 2 to 8 carbon atoms may be linear or branched and includes, for example, vinyl, propenyl, 2-butenyl, 3-butenyl, isobutenyl, 2-octenyl and the like. The alkynyl group of the above (3) is a lower alkynyl group having about 2 to 8 carbon atoms and may be linear or branched. For example, ethynyl, 2-propynyl, 2-butynyl, 2-pentynyl, 2-pentynyl, Octynyl and the like. Examples of the cycloalkyl group of the above (4) include lower cycloalkyl having about 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. The above alkyl group, alkenyl group, alkynyl group or cycloalkyl group is a hydroxyl group, an amino group which may be substituted (eg,
Amino, N-lower (C _1-4 ) alkylamino, N, N
-Di-lower (C _1-4 ) alkylamino, etc.), halogen, lower (C _1-4 ) alkoxy, lower (C _1-4 )
It may be substituted with an alkylthio group or the like. Examples of the aralkyl group in the above (5) include phenyl-lower (C _1-4 ) alkyl such as benzyl and phenethyl, and examples of the aryl group in the above (6) include phenyl. The aralkyl group or the aryl group described above may be, for example, halogen (eg, F, Cl, Br, etc.), nitro, or an optionally substituted amino group (eg, amino, N-lower) at any position on the benzene ring. (C _1-4 ) alkylamino, N, N-di-lower (C
_1-4 ) alkylamino, etc.), lower (C _1-4 ) alkoxy (eg, methoxy, ethoxy, etc.), lower (C
_1-4 ) alkylthio (eg, methylthio, ethylthio, etc.), lower (C _1-4 ) alkyl (eg, methyl, ethyl, etc.), and the like. Among the above, R
The “hydrocarbon residue” in the “hydrocarbon residue which may be bonded via a hetero atom and has a substituent” represented by ³ represents an optionally substituted alkyl or alkenyl group ( Examples are preferably a hydroxyl group, an amino group, a halogen or a lower (C _1-5 ) alkyl or a lower (C _2-5 ) alkenyl group which may be substituted with a lower (C _1-4 ) alkoxy group, and particularly, Lower (C _1-5 ) alkyl (more preferably, ethyl) is preferred. The “hetero atom” in the “hydrocarbon residue which may be bonded via a hetero atom and has a substituent” represented by R ³ includes —O—, —S (O) _m — [ m
Represents an integer of 0 to 2], -NR '-[R' represents a hydrogen atom or lower (C _1-4 ) alkyl], and among them, -O- is preferably used. Among the above-mentioned, as ^{R 3} is, -O -, - S (O ) m -
[M represents an integer of 0 to 2] or -NR'-
[R ′ represents a hydrogen atom or a lower (C _1-4 ) alkyl], and may be a hydroxyl group, an amino group,
A lower (C _1-5 ) alkyl or lower (C _2-5 ) alkenyl group which may be substituted with a substituent selected from a halogen and a lower (C _1-4 ) alkoxy group is preferred, and particularly, a lower (C _1-5 ) alkenyl group is preferred. _1-5 ) alkyl or lower ( _C1-5 ) alkoxy (more preferably ethoxy)
Is preferred.

Among the compounds represented by the formula (I), the compounds represented by the formula (I ')

Embedded image (Wherein R ¹ represents (1) a carboxyl group, (2) a tetrazolyl group, or (3)

Embedded image [Wherein, i represents -O- or -S-, j is> = O,
> = S or> = indicates S (O) _{m, m} is a group represented by as defined above], ring A may be substituted in addition to the substituent R ² a lower (C _{1 -4} ) alkyl (eg,
A hydroxyl group, a carboxyl group, a lower (C _1-4 ) alkyl which may be substituted with halogen, etc.) or a benzene ring which may be substituted with halogen (preferably having a substituent other than the substituent R ²⁾ No benzene ring)
R ² is wherein -CO-D [wherein, D is (1) hydroxy group or (2) alkyl moiety by hydroxyl, amino, halogen, lower _{(C 2 -6)} alkanoyloxy (e.g., acetoxy,
Pivaloyloxy, etc.), lower _{(C 3-8)} cycloalkanoyloxy, lower _{(C 1-6)} alkoxycarbonyloxy (e.g., methoxycarbonyloxy, such as ethoxycarbonyloxy), lower _{(C 3-8)} cycloalkyl alkoxycarbonyl Niro carboxymethyl (eg, such as cyclohexyloxycarbonyloxy), lower _{(C 1-4)} alkoxy or lower _{(C 3-8)} group represented by which may lower substituted with cycloalkoxy _{(C 1-4)} an alkoxy] R ³ represents -O-, -S (O) _m- [m represents an integer of 0 to 2] or -NR '-[R' represents a hydrogen atom or a lower (C _1-4 ) alkyl ] may be bonded via a hydroxy group, an amino group, substituted by halogen and lower (C _1-4) substituents selected from alkoxy groups Which may be a lower _{(C 1-5)} alkyl or lower _{(C 2-5)} alkenyl group (preferably a lower (C
_1-5 ) alkyl or lower ( _C1-5 ) alkoxy;
More preferably, ethoxy). A benzimidazole-7-carboxylic acid derivative or a pharmacologically acceptable salt thereof,
Ethoxy-1-[[2 ′-(1H-tetrazole-5-
Yl) biphenyl-4-yl] methyl] benzimidazole-7-carboxylic acid [Candesartan], 1- (cyclohexyloxycarbonyloxy) ethyl 2-ethoxy-1-[[2 '-(1H-tetrazol-5-yl)
Biphenyl-4-yl] methyl] benzimidazole-
7-carboxylate [Candesartan cilexetil], pivaloyloxymethyl 2-ethoxy-1-[[2'-
(1H-tetrazol-5-yl) biphenyl-4-yl] methyl] benzimidazole-7-carboxylate, 2-ethoxy-1-[[2 ′-(2,5-dihydro-5-oxo-1,2) , 4-oxadiazol-3-yl) biphenyl-4-yl] methyl] benzimidazole-7-carboxylic acid or a salt thereof is preferred. The above-mentioned benzimidazole derivatives are, for example, EP-42
5921, EP-459136, EP-55387
9, EP-578125, EP-520423,
It can be synthesized by a known method described in EP-668272 or the like or a method analogous thereto. When candesartan cilexetil is used, E
It is preferable to use a stable C-type crystal described in P-59136.

The amount of the physiologically active substance in the sustained-release preparation of the present invention varies depending on the kind of the physiologically active substance and the like.
(W / W), preferably about 0.2-30% (W / W),
More preferably, it is about 0.5 to 20% (W / W),
In the case of a non-peptide physiologically active substance, usually about 0.1 to 6
0% (W / W), preferably about 0.2-40% (W / W).
W), more preferably about 0.5 to 30% (W / W).

The polymer used in the present invention is a biopolymer which is hardly soluble or insoluble in water and is biocompatible. Examples of the polymer include polystyrene, polyacrylic acid, polymethacrylic acid, and the like. Copolymer of acrylic acid and methacrylic acid, nylon, tetron, silicone polymer, dextranstearate, ethylcellulose,
Examples include acetylcellulose, nitrocellulose, polyurethane, ethylene vinyl acetate copolymer, polyvinyl acetate, polyvinyl alcohol, polyacrylamide, and the like. Examples of the biodegradable polymer include, for example, α-hydroxycarboxylic acids (eg, glycolic acid, lactic acid, etc.), hydroxydicarboxylic acids (eg, malic acid, etc.), and hydroxytricarboxylic acids (eg, citric acid, etc.). Polymers having free carboxyl groups or mixtures thereof, poly-α-cyanoacrylates, polyamino acids (eg, poly-γ-benzyl-L-glutamic acid, etc.) synthesized from non-catalytic dehydration-condensation polymerization from at least one species And a maleic anhydride polymer (eg, a styrene-maleic acid polymer). These may be either homopolymers or copolymers. The type of polymerization may be random, block, or graft. When the α-hydroxycarboxylic acids, hydroxydicarboxylic acids, and hydroxytricarboxylic acids have an optically active center in the molecule, any of D-, L-, and DL-forms can be used. Among these, biodegradable polymers having a terminal free carboxyl group, for example, α-
Hydroxycarboxylic acids (eg, glycolic acid, lactic acid, etc.)
(Eg, lactic acid polymers, lactic acid-glycolic acid copolymers, etc.), and poly-α-cyanoacrylates are preferred. The biodegradable polymer is more preferably a polymer synthesized from α-hydroxycarboxylic acids, particularly preferably a lactic acid-glycolic acid polymer.

In this specification, lactic acid-glycolic acid polymer may be simply referred to as lactic acid-glycolic acid copolymer, as well as a homopolymer such as polylactic acid and polyglycolic acid. When a lactic acid-glycolic acid polymer (lactic acid-glycolic acid copolymer or homopolymer) is used as the biodegradable polymer, its composition ratio (mol%) is about 10%.
0/0 to about 40/60 are preferred, and about 85/15 to about 5
0/50 is more preferred. The weight average molecular weight of the lactic acid-glycolic acid polymer is preferably about 3,000 to about 50,000, more preferably about 3,000 to about 25,000, and still more preferably about 5,000 to about 20,000.
Degree of dispersion of lactic acid-glycolic acid polymer (weight average molecular weight /
(Number average molecular weight) is preferably about 1.2 to about 4.0, and more preferably about 1.5 to about 3.5.

As for the weight average molecular weight and the degree of dispersion in the present specification, the former has a weight average molecular weight of 12,000,
0,52,000,22,000,9,200,5,0
A value in terms of polystyrene measured by gel permeation chromatography (GPC) using nine kinds of polystyrenes of 50, 2,950, 1,050, 580, and 162 as a reference substance, and the latter is a value calculated from this value. The measurement is performed using a GPC column KF804Lx2 (manufactured by Showa Denko) and an RI monitor L-3300 (manufactured by Hitachi, Ltd.) using chloroform as a mobile phase.

The term "biodegradable polymer having a free carboxyl group at the terminal" refers to a polymer having a number average molecular weight almost identical to the number average molecular weight determined by the above-described GPC measurement, and the number average molecular weight determined by the above-described GPC measurement. The average molecular weight is calculated as follows. About 1 g to about 3 g of a biodegradable polymer is dissolved in a mixed solvent of acetone (25 ml) and methanol (5 ml), and phenolphthalein is used as an indicator to reduce carboxyl groups in the solution to 0.03 g.
5N alcoholic potassium hydroxide solution at room temperature (20
C.), and the mixture was rapidly titrated under stirring, and the number average molecular weight determined by terminal group quantification was calculated by the following equation. Number average molecular weight by terminal group determination = 20,000 × A / B A: Mass of biodegradable polymer (g) B: 0.05N alcoholic potassium hydroxide solution (ml) added up to the end of titration Number by terminal group determination Whereas the average molecular weight is an absolute value, the number average molecular weight measured by GPC can be determined by various analysis and analysis conditions (eg, mobile phase type, column type, reference material, selection of slice width, selection of baseline, etc.). ), It is difficult to obtain a unique numerical value, but it is considered that the number average molecular weights obtained by both measurements are almost the same, for example, in a polymer synthesized from α-hydroxycarboxylic acids. The number average molecular weight determined by quantification is in the range of about 0.5 times to about 2 times, preferably in the range of about 0.7 times to about 1.5 times, the number average molecular weight determined by GPC. That means.

For example, in the case of a polymer synthesized from one or more α-hydroxycarboxylic acids by a noncatalytic dehydration condensation polymerization method and having a free carboxyl group at a terminal, the number average molecular weight by GPC measurement and the number average molecular weight by terminal group quantification The molecular weights are almost the same. On the other hand, in a polymer synthesized from a cyclic dimer by a ring-opening polymerization method using a catalyst and having essentially no free carboxyl group at the terminal, the number average molecular weight determined by the terminal group is determined by the GPC measurement. Significantly more than about twice the average molecular weight. By this difference, a polymer having a free carboxyl group at a terminal can be clearly distinguished from a polymer having no free carboxyl group at a terminal.

A lactic acid-glycolic acid polymer having a free carboxyl group at a terminal can be produced by a method known per se, for example, a method described in JP-A-61-28521 (for example, a dehydration-condensation polymerization reaction or an inorganic polymerization reaction without a catalyst). A dehydration-condensation polymerization reaction in the presence of a solid acid catalyst). The decomposition / elimination rate of the lactic acid-glycolic acid polymer greatly varies depending on the composition ratio or the weight average molecular weight.
Due to the slow elimination, the release period can be extended (eg, about 6 months) by lowering the glycolic acid fraction or increasing the molecular weight. Conversely, the release period can be shortened (eg, about one week) by increasing the glycolic acid fraction or decreasing the molecular weight. For example, in order to prepare a one-week to two-month sustained release preparation, it is preferable to use a lactic acid-glycolic acid polymer having the above composition ratio and weight average molecular weight.

Therefore, it is preferable that the composition of the biodegradable polymer used in the present invention is appropriately selected depending on the kind of the target physiologically active substance, a desired sustained release period, and the like. As a specific example, for example, when GH is used as a physiologically active substance, it is preferable to use a lactic acid-glycolic acid polymer, and the lactic acid / glycolic acid polymer has a lactic acid / glycolic acid composition ratio (mol%). Is 8
A lactic acid-glycolic acid copolymer of 5/15 to about 50/50 is preferred, more preferably about 75/25 to about 50/50.
50 lactic acid-glycolic acid copolymer. The weight average molecular weight is preferably from about 8,000 to about 20,000, more preferably from about 10,000 to about 20,000.
It is. The degree of dispersion (weight average molecular weight / number average molecular weight) of the lactic acid-glycolic acid polymer is preferably from about 1.2 to about 4.0, more preferably from about 1.5 to about 3.5.

The lactic acid-glycolic acid polymer to be used can be produced according to a known method such as the method described in the above publication. The polymer is preferably produced by non-catalytic dehydration-condensation polymerization.
The organic solvent used in the production of the polymer and remaining in the polymer is removed after the polymerization reaction. Examples of the method include, for example, a heat drying method, a vacuum drying method, a flash drying method using a dry gas, or a method similar thereto. It can be removed quickly and the time required for solvent removal is greatly reduced. It is preferable to use a lactic acid-glycolic acid polymer (PLGA) in which the number average molecular weight by the GPC measurement method and the number average molecular weight by the terminal group quantification method are almost the same. The polymer has a composition ratio and / or
Alternatively, two kinds of lactic acid-glycolic acid polymers having different weight average molecular weights may be mixed and used at an arbitrary ratio. As such an example, the composition ratio (lactic acid / glycolic acid) (mol%) is about 75/25 and the weight average molecular weight is about 10,000.
And a lactic acid-glycolic acid copolymer having a composition ratio (lactic acid / glycolic acid) (mol%) of about 50/50 and a weight average molecular weight of about 12,000. Can be The weight ratio upon mixing is preferably from about 25/75 to about 75/25.

The biodegradable polymer used in the present invention may be a metal salt of the above-described biodegradable polymer. A valent metal salt or the like is used. Preferably, a polyvalent metal salt of a lactic acid-glycolic acid polymer (further preferably a zinc salt, a calcium salt, a magnesium salt or the like, more preferably a zinc salt or the like) is used. The metal species of the polyvalent metal salt is not particularly limited as long as it is a compound that does not adversely affect the living body.
Polyvalent metals such as iron, zinc, copper, calcium, magnesium, aluminum, tin, and manganese), trivalent (eg, iron, aluminum, manganese, and the like) and tetravalent (eg, tin and the like) can also be used.

In the present specification, the biodegradable polymer may be referred to as a biodegradable polymer including the case where the biodegradable polymer is a metal salt. It may be referred to as a glycolic acid polymer. These polyvalent metal salts of a biodegradable polymer can be produced by the method described in WO 97/01331 or a method analogous thereto. When the polyvalent metal salt of the biodegradable polymer is a zinc salt, it can be produced by reacting the biodegradable polymer with zinc oxide in an organic solvent.

In the production method, first, a biodegradable polymer and zinc oxide are allowed to coexist in an organic solvent to prepare a biodegradable polymer / zinc oxide solution in an organic solvent. At this time, the concentration of the biodegradable polymer in the solution varies depending on the molecular weight, the type of the organic solvent, and the like.
1 to about 80% (W / W), preferably about 1 to about 70%
(W / W), more preferably about 2 to about 60% (W / W).
W). Further, the amount of zinc oxide to be added was
As described in JP-A-231252, although it differs depending on the type of the organic solvent, for example, when the target physiologically active substance is a peptide, the amount of the biodegradable polymer is about 0.0
01 to about 2% (W / W), preferably about 0.01 to about 1.5% (W / W), more preferably about 0.1 to about 1
% (W / W), and when the target physiologically active substance is a non-peptide, the amount of the biodegradable polymer is about 0.001 to 0.001%.
About 30% (W / W), preferably about 0.01 to about 20%
(W / W), more preferably about 0.1 to about 10% (W / W).
/ W). The order of adding the biodegradable polymer and the zinc oxide to the organic solvent may be such that zinc oxide may be added to the solution of the biodegradable polymer in the organic solvent in the form of powder or suspended in the organic solvent. An organic solvent solution of a biodegradable polymer may be added to a suspension of zinc oxide in an organic solvent. Further, an organic solvent may be added after mixing both in powder form. When the target physiologically active substance is a non-peptide, an organic solvent may be added after mixing the biodegradable polymer, zinc oxide and the physiologically active substance in powder form.

The content of the biodegradable polymer contained in the preparation of the present invention is usually about 30 to 99.9% (W / W), preferably about 60 to 97% (W / W), and more preferably about 60 to 97% (W / W). 70-90% (W / W).

The preparation of the present invention is produced by first forming a solid containing a physiologically active substance and a biodegradable polymer, and then contacting the solid with a high-pressure gas. The solid containing the bioactive substance and the biodegradable polymer is, for example, when the bioactive substance is a bioactive peptide, a powder obtained by freeze-drying a bioactive peptide solution (S
Phase) was dissolved in an organic solvent solution (O
Phase), by removing the solvent from the S / O type dispersion, or by dissolving the biodegradable polymer in the aqueous phase (W phase) in which the physiologically active peptide is dissolved in an organic solvent (O phase). ), By removing the solvent from the W / O emulsion, or by removing the solvent from a solution in which a physiologically active peptide is dissolved in an organic solvent solution (O phase) together with a biodegradable polymer. ,It is formed. Examples of the method include (a) an underwater drying method (S / O / W
Method and W / O / W method or O / W method), (b) phase separation method (coacervation method) and (c) spray drying method, or a method analogous thereto. In this specification, a solid substance refers to a substance in which its constituent substances are physically or chemically bonded. The solid material is not particularly limited, but includes microcapsules and the like.

The organic solvent used for dissolving the biodegradable polymer preferably has a boiling point of 30 ° C. or higher. Examples of the organic solvent include halogenated hydrocarbons such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride, pentane, hexane, heptane, cyclohexane, petroleum benzine, petroleum hydrocarbons such as petroleum ether, toluene, xylene and the like. Aromatic hydrocarbons, alcohols such as methyl alcohol, ethyl alcohol, n-propyl alcohol, isopropyl alcohol, and benzyl alcohol; polyhydric alcohols such as ethylene glycol and propylene glycol; esters such as methyl acetate and ethyl acetate; Acetic acid, trifluoroacetic acid, organic acids such as trichloroacetic acid, diethyl ether, isopropyl ether, dioxane, ethers such as tetrahydrofuran, acetone, ketones such as methyl ethyl ketone, Acetonitrile, propionitrile, pyridine, dimethylacetamide, nitrogen-containing compounds such as dimethyl formamide, sulfur-containing compounds such as dimethyl sulfoxide and the like. These may be mixed and used at an appropriate ratio. Substances, especially organic compounds,
For example, the solvent contained in a drug must be substantially removed from the product due to the properties of the drug. Further, solvents remaining in products of foods and general chemicals are strictly regulated depending on the use. Regarding the amount of residual solvent in pharmaceutical products, guidelines based on ICH (“Guidelines for residual solvent in pharmaceutical products”, Pharm.Tech.Japan 16
(5), 687-704, 2000) has a description of the allowable value.
The concentration limit value of acetone classified as 0 ppm and class 3 is 0.5% (5,000 ppm).

The organic solvent used for dissolving the biodegradable polymer preferably has a boiling point of 120 ° C. or lower. Examples of the organic solvent include halogenated hydrocarbons (eg, dichloromethane, chloroform, etc.), alcohols (eg, ethanol, methanol, etc.), ethyl acetate, acetonitrile, and the like. These may be mixed and used at an appropriate ratio. When an organic solvent is used alone, for example, dichloromethane, ethyl acetate, acetonitrile and the like are preferable. When an organic solvent is used as a mixed solvent, for example, a combination of a halogenated hydrocarbon (eg, dichloromethane, chloroform, etc.) with an alcohol (eg, ethanol, methanol, etc.) or acetonitrile is preferred. The mixing ratio (volume ratio) between the halogenated hydrocarbon and the alcohol or acetonitrile is about 100: 1 to about 1: 1, and preferably a mixed solvent of about 30: 1 to about 2: 1 is used. . The concentration of the biodegradable polymer in the solution varies depending on the molecular weight, the type of the organic solvent, and the like. / W), more preferably about 1 to about 60% (W / W).

The method of microencapsulation when a sustained release microcapsule is produced as a preparation using a physiologically active peptide as a physiologically active substance will be described in more detail below.

(A-1) Underwater drying method (S / O / W method) According to this method, a water-miscible organic solvent and / or volatile salts are first added to a physiologically active peptide aqueous solution, followed by freeze-drying. To prepare a physiologically active peptide powder (S phase). Next, the biodegradable polymer is dissolved in an organic solvent, and the above-mentioned physiologically active peptide powder is added and dispersed in the organic solvent liquid. At this time, the ratio (weight ratio) of the bioactive peptide to the biodegradable polymer is, for example, about 1: 1000.
To about 1: 1, preferably about 1: 200 to about 1: 5, more preferably about 1: 100 to about 1: 5. In order to uniformly disperse the bioactive peptide powder in the organic solvent liquid, it is preferable to apply external physical energy.
As the method, for example, ultrasonic irradiation, a turbine type stirrer, a homogenizer and the like are used. At this time, the average particle size of the physiologically active peptide in the organic solvent solution is about 1
0 μm or less, more preferably about 0.1 μm to about 10 μm
m, more preferably about 0.5 μm to 5 μm. The average particle size of the physiologically active peptide in the present invention is determined by dispersing the physiologically active peptide in an organic solvent such as dichloromethane using a homogenizer, and then using a laser analysis type particle size distribution analyzer (SALD2000A: Shimadzu).
Shows the value obtained by At that time, the bioactive peptide is added to an organic solvent such as dichloromethane, for example, about 20 to 100.
After adding at a concentration of mg / ml, using a homogenizer (eg, Polytron (Kinematica)) to about 20,000 rpm
The mixture is stirred for about 30 seconds to 1 minute at m to form a dispersion liquid, and then appropriately diluted with the organic solvent so as to be within the measurable range of the particle size distribution measuring device described above.

Next, the organic solvent dispersion (S / O type dispersion) thus prepared was further added to an aqueous solvent (W phase).
To form an S / O / W emulsion by the same external physical energy as described above, for example, by ultrasonic irradiation, a turbine stirrer, or a homogenizer. Thereafter, the oil phase solvent is evaporated to produce microcapsules. The volume of the water phase at this time is generally about 1 to the volume of the oil phase.
Double to about 10,000 times. More preferably, it is selected from about 2 times to about 5,000 times. It is particularly preferably selected from about 5 times to about 2,000 times.

An emulsifier may be added to the outer aqueous phase. The emulsifier may be any as long as it can form a generally stable S / O / W emulsion. Examples of the emulsifier include anionic surfactants, nonionic surfactants, polyoxyethylene castor oil derivatives, polyvinylpyrrolidone, polyvinyl alcohol, carboxymethylcellulose, lecithin, gelatin, hyaluronic acid, and the like. These may be used in appropriate combination. The concentration of the emulsifier in the external aqueous phase is preferably about 0.0
01% to 20% (W / W). More preferably about 0.01% to 10% (W / W), particularly preferably about 0.1% to 10% (W / W).
05% to 5% (W / W). The microcapsules thus obtained are separated by centrifugation or filtration, and then the emulsifiers and the like adhering to the surface of the microcapsules are removed by washing with distilled water and dispersed again in distilled water or the like. Lyophilize. Examples of the water-miscible organic solvent to be added to the aqueous physiologically active peptide solution in the present method include alcohols (eg, methanol, ethanol, isopropanol, etc., preferably methanol, ethanol, etc.), acetone and the like. These may be mixed and used at an appropriate ratio, but it is preferable to use alcohols, particularly ethanol alone. The addition amount (concentration) to the aqueous solution of the physiologically active peptide is about 0.03 to 0.5% (V / V) by volume, preferably about 0.06 to 0.25% (V / V). ), More preferably about 0.1 to 0.15% (V / V). The aqueous solution of the physiologically active peptide obtained by adding such a water-miscible organic solvent is further freeze-dried, so that it is easy to handle (has good operability) and is fine (small particle size). Powder can be made. Examples of the volatile salts added to the aqueous physiologically active peptide solution in the present method include ammonium salts (for example, ammonium acetate, ammonium bicarbonate, ammonium carbonate, ammonium chloride, and the like, preferably ammonium acetate and the like). These may be mixed and used at an appropriate ratio. The amount of the volatile salt to be added to the aqueous physiologically active peptide solution is about 10-fold to about 80-fold by mole, preferably about 10-fold to about 70-fold, and more preferably about 15-fold to about 70-fold. The molar amount is 70 times, more preferably about 20 times to about 70 times, and most preferably about 20 times to about 50 times. As in the case of adding a water-miscible organic solvent, the aqueous solution of a physiologically active peptide obtained by adding a volatile salt is further freeze-dried, so that it is easy to handle (has good operability) and has a fine ( A fine bioactive peptide powder (with a small particle size) can be prepared. In this method, the water-miscible organic solvent and / or volatile salts added to the aqueous physiologically active peptide solution may be used alone or in an appropriate combination. When a water-miscible organic solvent and volatile salts are used in combination, they can be added to the aqueous physiologically active peptide solution according to the respective addition amounts described above.

(A-2) Underwater drying method (W / O / W method) According to this method, first, water or an appropriate buffer is added to a physiologically active peptide to prepare a physiologically active peptide solution (W phase). I do. Next, the biodegradable polymer is dissolved in an organic solvent,
The above-mentioned physiologically active peptide solution is added to and dispersed in this organic solvent solution. The W / O emulsion thus obtained is further added to an aqueous solvent (W phase), and the microcapsules are passed through a W / O / W emulsion in the same manner as in the above S / O / W method. Get.

(A-3) Underwater drying method (O / W method) According to this method, first, a biodegradable polymer is dissolved in an organic solvent together with a physiologically active peptide, and an organic solvent solution (O phase) is dissolved.
Is further added to an aqueous solvent (W phase), and the above S / O /
Microcapsules are obtained via an O / W emulsion as in the W method.

(B) Phase separation method (coacervation method) In the present method, the S / O type dispersion of (a-1)
A coacervating agent is gradually added to the W / O emulsion (a-2) or the oil phase solution (a-3) with stirring to precipitate and solidify microcapsules. The coacervation agent is added in a volume of about 0.01 to about 1,000 times the volume of the dispersion. More preferably, about 0.05 times to about 500 times.
Times, particularly preferably about 0.1 to about 200 times the volume. The coacervation agent may be any as long as it does not dissolve the biodegradable polymer used in the polymer, mineral oil or vegetable oil-based compound that is miscible with the organic solvent that dissolves the biodegradable polymer. Specifically, for example, silicon oil, sesame oil, soybean oil, corn oil, cottonseed oil, coconut oil, linseed oil, mineral oil, n-hexane, n-heptane and the like are used. These may be used as a mixture of two or more. After the microcapsules thus obtained are collected by filtration, they are repeatedly washed with heptane or the like to remove the coacervation agent. Further, it is washed and freeze-dried in the same manner as in (a) above. In the production of microcapsules by the in-water drying method and the coacervation method, an aggregation inhibitor may be added to prevent aggregation of particles. Examples of the coagulation inhibitor include water-soluble polysaccharides such as mannitol, lactose, glucose, starches (eg, corn starch), hyaluronic acid and alkali metal salts thereof, proteins such as glycine, fibrin, collagen, sodium chloride, and phosphoric acid. Inorganic salts such as sodium hydrogen and the like are appropriately used.

(C) Spray drying method In the present method, the S / O type dispersion of (a-1), the W / O type emulsion of (a-2) or the oil phase solution of (a-3) With a nozzle using a spray dryer (spray dryer)
And the organic solvent in the atomized droplets is volatilized in a very short time to produce microcapsules. As the nozzle, for example, a two-fluid nozzle type, a pressure nozzle type,
There is a rotating disk type and the like. In this case, if desired, it is also effective to spray an aqueous solution of the above-mentioned anti-agglomeration agent from another nozzle simultaneously with the above-mentioned dispersion liquid for the purpose of preventing the micro-capsule particles from agglomerating.

The solid formed by the above method, such as microcapsules containing a physiologically active substance and a biodegradable polymer, is subsequently contacted with a high-pressure gas (preferably carbon dioxide) to further extract and remove the organic solvent. I do.

Specifically, for example, the freeze-dried microcapsule powder obtained in (a) is placed in an extraction container, and an extraction process is performed using an extraction system including a carbon dioxide feed pump and a pressure control valve. It is also possible to put the microcapsule suspension before freeze-drying obtained in (a) or (b) in an extraction container and perform the extraction treatment in the same manner. In these cases, because the quality of the sustained release formulation is not impaired,
It is preferable to perform the extraction process under more relaxed conditions.

The high-pressure gas in the present invention is a gas whose pressure is equal to or higher than atmospheric pressure at a certain temperature and equal to or lower than the liquefaction pressure at that temperature. Examples of the high-pressure gas used in the present invention include carbon dioxide, nitrous oxide, nitrogen, helium, argon, alkanes (eg, ethane, propane, etc.), and alkenes (eg, ethylene, etc.). These may be used in a mixture at an appropriate ratio, but it is preferable to use carbon dioxide alone. If the temperature of the high-pressure gas in contact with the preparation is too high beyond the glass transition temperature of the biodegradable polymer used as the base of the preparation, fusion of the preparation,
The risk of deformation, decomposition and degradation of the physiologically active substance increases. The glass transition temperature in the present invention refers to a midpoint glass transition temperature obtained when the temperature is raised at a heating rate of 10 or 20 ° C. per minute using a differential scanning calorimeter (DSC). If the temperature of the high-pressure gas is too low, the removal of the organic solvent will be insufficient. The organic solvent is less than 1,000 ppm,
Preferably less than 500 ppm, more preferably 100p
It is better to remove to less than about pm. Therefore, the advantageous temperature when carbon dioxide is used as the high-pressure gas in the present invention is +20 to -60 ° C based on the glass transition temperature of the biodegradable polymer (usually about 20 to 60 ° C),
Preferably +10 to -50 ° C, more preferably 0 to -4
The temperature range is 0 ° C, more preferably -5 to -30 ° C, and most preferably -10 to -25 ° C.

Although the range of pressure during use differs depending on the selected high-pressure gas, generally, if the pressure of the high-pressure gas is too high, there is a risk that the microcapsules are fused, deformed, and the initial release immediately after administration is increased. If the pressure is too high or the pressure is too low, the removal of the organic solvent will be insufficient. Advantageous pressures when using carbon dioxide as a high-pressure gas in the present invention are about 1 to 7 MPa,
Preferably it is about 1 to 4 MPa, more preferably about 2 to 4 MPa. The time of contact with the high-pressure gas also varies depending on the pressure, temperature, amount of microcapsules to be processed, etc.
When using carbon dioxide as high pressure gas, it takes about 5 minutes
About 48 hours are preferred. More preferably, about 10 minutes to about 12 hours.

Hereinafter, the high-pressure gas treatment step of microcapsules using carbon dioxide in a high-pressure gas state will be described in more detail with reference to FIG. FIG. 1 is a schematic view of an example of an apparatus used for high-pressure gas processing in the present invention. As shown in FIG. 1, for example, such a high-pressure gas processing apparatus includes a liquefied carbon dioxide cylinder 1, a carbon dioxide liquid sending pump 2, a heat exchanger 3, an extraction vessel 4, a constant temperature bath 5, a detector 6, a fully automatic pressure regulating valve 7, It is composed of a collection container 8. The microcapsules to be treated are placed in the extraction container 4 and after closing the device,
It is heated to a predetermined temperature. Next, the liquefied carbonic acid is sent from the liquefied carbonic acid cylinder 1 to the heat exchanger 3 by the carbon dioxide liquid sending pump 2, heated to a predetermined temperature, and converted into a high-pressure gas state. Next, the carbon dioxide in the high-pressure gas state is blown into the extraction container 4, and the solvent in the microcapsules is dissolved and extracted in the high-pressure gas. The extracted solvent is detected by the detector 6,
It is collected in the collection container 8 via the automatic pressure control valve 7. The pressure applied to the entire system is controlled by a fully automatic pressure regulating valve 7 connected at the most downstream. By contacting with the high-pressure gas for a predetermined time, the initial release of an excessive amount of the physiologically active substance immediately after administration is drastically suppressed, and without generating a polymer, an analogous substance or a reactant of the physiologically active peptide. , The residual organic solvent can be removed.

The sustained-release preparation of the present invention is preferably in the form of fine particles. This is because the sustained release preparation does not cause undue pain to the patient when administered through a needle usually used for subcutaneous or intramuscular injection. The particle size of the sustained-release preparation is, for example, about 0.1 to 300 μm as an average particle diameter, preferably about 1 to 150 μm,
Particularly preferably, it is about 2 to 100 μm. The content of the physiologically active substance contained in the sustained-release preparation of the present invention is usually, for example, about 0.1 to 50% (W / W) for a physiologically active peptide,
Preferably it is about 0.2-30% (W / W), more preferably about 0.5-20% (W / W). The content of the biodegradable polymer contained in the sustained-release preparation of the present invention is usually about 30 to 99.9% (W / W), preferably about 60 to 9%.
7% (W / W), more preferably about 70-90% (W / W)
/ W). The initial release rate [the release rate up to one day (24 hours) after administration] of the sustained-release preparation of the present invention is preferably about 40% or less, more preferably about 1 to 40%, in the case of a bioactive peptide. Preferably it is about 3-35%.

The sustained-release preparation of the present invention can be used as a parenteral preparation (eg, intramuscular, subcutaneous, organ, etc.) as a microcapsule or a preparation prepared by using the microcapsule as a raw material in various dosage forms. Solid preparations such as injections or implants, transmucosal preparations for the nasal cavity, rectum, uterus, etc.), oral preparations (eg, capsules (eg, hard capsules, soft capsules, etc.), granules, powders, etc.) Liquids such as suspensions)
Etc. can be administered. The preparation of the present invention is particularly preferably for injection. For example, when the sustained-release preparation is a microcapsule, the microcapsule is used as a dispersant (eg, a surfactant such as Tween 80 or HCO-60, a polysaccharide such as carboxymethylcellulose, sodium alginate, or hyaluronic acid), and a preservative. (Examples: methylparaben, propylparaben, etc.) and isotonic agents (eg, sodium chloride, mannitol, sorbitol, glucose, etc.) to form an aqueous suspension, whereby a practical sustained-release injection can be obtained. In addition, vegetable oils such as sesame oil and corn oil or a mixture thereof with a phospholipid such as lecithin, or dispersed with medium-chain fatty acid triglyceride (eg, miglyol 812), can be used as an oily suspension for sustained release injection. Agent.

When the sustained-release preparation is, for example, a microcapsule, the particle size of the microcapsule may be within a range satisfying the degree of dispersion and needle penetration when used as a suspension injection. The average particle size ranges from about 0.1 to about 300 μm. The average particle size is preferably about 1 to about 150 μm, particularly preferably about 2 to about 10 μm.
The range is 0 μm. In order to make the above-mentioned microcapsules into a sterile preparation, a method of sterilizing the whole production process, a method of sterilizing with gamma rays, a method of adding a preservative, and the like are mentioned, but are not particularly limited.

The sustained-release preparation of the present invention has low toxicity and can be safely used for mammals (eg, human, cow, pig, dog, cat, mouse, rat, rabbit, etc.). The application of the sustained-release preparation varies depending on the physiologically active substance used.
When the physiologically active substance is, for example, insulin, diabetes, etc., and when it is interferon-α, viral hepatitis (eg, hepatitis C, HBe antigen-positive active hepatitis, etc.), cancer (eg, renal cancer) Erythropoietin, anemia (eg, anemia during renal dialysis), G-
In the case of CSF, neutropenia (eg, during anticancer drug treatment), infectious disease, etc., in the case of IL-2, cancer (eg, hemangioendothelioma), etc., in the case of FGF, fracture, wound (bedsore) etc),
Periodontal disease, gastrointestinal ulcer, etc., thrombocytopenia etc. for FGF-9, senile dementia, neuropathy etc. for NGF, thrombosis etc. for TPA, tumor necrosis factor In this case, it is effective for treatment or prevention of cancer and the like. In the GH-containing sustained-release preparation, GH secretion deficient short stature (pituitary dwarfism) is based on the growth hormone action of GH.
In addition, the present invention can be applied to the treatment of depleting diseases such as Turner's syndrome, chronic kidney disease, cartilage dysgenesis (chondrodystrophy), adult growth hormone deficiency (adult GHD), and AIDS. It has also been reported that GH has been applied to diseases such as Down syndrome, Silver syndrome, osteogenesis imperfecta, or juvenile chronic arthropathy, and has obtained an effective therapeutic effect. It is also adaptable to diseases. It is also effective for treating or preventing congestive heart failure and the like. Other subjects to which the GH-containing sustained-release preparation is applicable include hematopoiesis during organ transplantation and drug treatment of AIDS patients,
Improve malnutrition, renal anemia, angina, hyperlipidemia, obesity, promote treatment of burns / wounds / ulcers, surgical invasion (surgery / trauma) / early recovery after surgery, sepsis, prevention of osteoporosis fractures ,
Examples include early recovery of postoperative muscular strength in patients with fractures due to osteoporosis, amyotrophic lateral sclerosis (ALS), and pressure ulcers. In addition, as an anti-aging drug for the purpose of improving the quality of life (QOL) of frail elderly people, or by hGH's neuroprotective effect, it is used to suppress and improve progression of neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, cerebrovascular disease, etc.) Can also be expected to be effective.
By formulating a sustained-release preparation of GH, superior drug efficacy can be obtained for these indications compared to daily GH subcutaneous injection. When the bioactive substance is candesartan,
Cardiac hypertrophy, heart failure, myocardial infarction, stroke, ischemic peripheral circulation disorder, myocardial ischemia, venous insufficiency, progression of heart failure after myocardial infarction, diabetic nephropathy, nephritis, glomerulonephritis, arteriosclerosis, vascular hypertrophy, Vascular thickening or occlusion after percutaneous coronary angioplasty,
Revascularization after bypass surgery, hyperaldosteronism, glomerulosclerosis, renal failure, glaucoma, ocular hypertension, hyperlipidemia, angina, aneurysm, coronary atherosclerosis, cerebral atherosclerosis, peripheral atherosclerosis Disease, thrombosis, central nervous system disease, Alzheimer's disease,
Prevention and treatment of memory deficiency, depression, amnesia, senile dementia, sensory dysfunction, multiple organ dysfunction, diseases or scleroderma associated with endothelial dysfunction, or anxiety, tension, discomfort, or indigestion It is effective for prevention and improvement.

The dosage of the sustained-release preparation varies depending on the type and content of the physiologically active substance, the duration of release, the target disease, the target animal, etc., but the amount at which the effective concentration of the physiologically active substance is maintained in the body. Is good enough. As the dose of the physiologically active substance, for example, when the sustained-release preparation is a two-week preparation,
Preferably about 0.0001 to about 10 mg per adult
/ Kg body weight range. More preferably, it can be appropriately selected from the range of about 0.05 to about 3 mg / kg body weight. The number of administrations is once a week, once every two weeks, once a month, once every two months, etc.
It can be appropriately selected depending on the target animal and the like. Preferably, a one week to two month type sustained release preparation is used, and more preferably, a one week to one month type sustained release preparation is used. When the physiologically active substance which is the active ingredient of the sustained-release preparation is, for example, insulin, the dosage for an adult with diabetes is usually about 0.001 to about 1 mg / kg body weight, preferably about 0 to about 1 mg / kg body weight as the active ingredient. The dose is suitably selected from the range of 0.01 to about 0.2 mg / kg body weight, and is preferably administered once a week.

When the physiologically active substance which is the active ingredient of the sustained-release preparation is GH, the dosage varies depending on the type and content of GH, the duration of release, the target disease, the target animal and the like. It is sufficient that the effective concentration of is maintained in the body. In the treatment of the above-mentioned diseases, for example, when the sustained-release preparation is a two-week preparation, the dose of GH as an active ingredient is preferably about 0.03 per pediatric or adult.
1 to about 5 mg / kg body weight (about 0.03 to about 15 IU / k
g body weight) and can be safely administered. More preferably, about 0.05 to about 1 mg / kg
It can be appropriately selected from the range of body weight (about 0.15 to about 3 IU / kg body weight). The frequency of administration can be appropriately selected depending on the GH content, dosage form, duration of release, target disease, target animal, etc., such as once a week, once every two weeks or once a month. Preferably, a one week to two month type sustained release preparation is used, and more preferably, a one week to one month type sustained release preparation is used. The sustained-release preparation is preferably stored at room temperature or in a cold place. More preferably, the sustained-release preparation is stored in a cold place. The normal temperature or the cold place here is defined in the Japanese Pharmacopoeia. That is, normal temperature means 15 to 25 ° C, and cold place means 15 ° C or less. Among cold places, 2 to 8C is particularly preferable.

[0065]

The present invention will be described in more detail with reference to the following Reference Examples, Examples and Test Examples, which do not limit the present invention. Reference Example 1 Genetically modified hGH aqueous solution (final hGH concentration = 2 mg)
/ Ml), 20 ml molar equivalent of ammonium acetate was added thereto, and 100 ml was dropped into the inner wall surface of the eggplant-shaped flask cooled in a dry ice-ethanol bath using a peristaltic pump for 30 minutes. By drying, hGH powder was obtained. Lactic acid-glycolic acid copolymer (lactic acid / glycolic acid = 65/35, viscosity = 0.160)
(dl / g) 1.690 g and zinc oxide 10 mg were dissolved in 2.7 ml of dichloromethane. 359 mg of the above hGH powder was added to this organic solvent liquid, and the mixture was pulverized using Polytron (Kinematica). This S / O dispersion was added to 0.
Add to 800ml of 1% polyvinyl alcohol aqueous solution,
The mixture was stirred and emulsified using a homomixer. After stirring at room temperature for 3 hours to evaporate dichloromethane, microcapsules were collected by centrifugation (about 1,500 rpm). Next, after washing twice with 400 ml of distilled water, 0.2 g of D-mannitol was added and freeze-dried to obtain freeze-dried microcapsule powder containing hGH. Producing 6 batches of microcapsules under the same conditions,
The yield of the obtained freeze-dried microcapsule powder was 6.8.
g.

Reference Example 2 A recombinant hGH aqueous solution (final hGH concentration = 2 mg)
/ Ml) was added with 20-fold molar equivalent of ammonium acetate and freeze-dried to obtain hGH powder. Lactic acid-
Glycolic acid copolymer (lactic acid / glycolic acid = 50/5
1.850 g (0, viscosity = 0.154 dl / g) and 10 mg of zinc oxide were dissolved in 2.7 ml of dichloromethane.
155 mg of the above hGH powder was added to this organic solvent liquid, and the mixture was pulverized using Polytron (Kinematica).
This S / O dispersion was added to 800 ml of a 0.1% aqueous solution of polyvinyl alcohol, and the mixture was stirred and emulsified using a homomixer. After stirring at room temperature for 3 hours to evaporate dichloromethane, microcapsules were collected by centrifugation (about 1,500 rpm). Then 400m of distilled water
After washing twice with l, 0.2 g of D-mannitol was added and freeze-dried to obtain freeze-dried microcapsule powder containing hGH. Six batches of microcapsules were produced under the same conditions, and the yield of the freeze-dried microcapsule powder obtained was 7.6 g.

Reference Example 3 Genetically modified hGH aqueous solution (final hGH concentration = 2 mg / m
To l), add a 20-fold molar equivalent of ammonium acetate,
Using a peristaltic pump, 100 ml was dropped into the inner wall surface of the eggplant-shaped flask cooled in a dry ice-ethanol bath for 30 minutes, rapidly frozen, and dried under vacuum to obtain hGH powder. Lactic acid-glycolic acid copolymer (lactic acid / glycolic acid = 65/35, viscosity = 0.160 dl /
g) 1.521 g and 9 mg of zinc oxide were dissolved in 2.4 ml of dichloromethane. The hGH described above is added to this organic solvent liquid.
Add 270mg of powder and add Polytron (Kinematica)
It was atomized by using. This S / O dispersion was added to 800 ml of a 0.1% aqueous polyvinyl alcohol solution cooled to 18 ° C., and the mixture was stirred and emulsified using a homomixer. After stirring at room temperature for 3 hours to evaporate dichloromethane, microcapsules were collected by centrifugation (about 1,500 rpm). After removing the supernatant as much as possible by suction, 500 ml of a 50% aqueous ethanol solution was added to the microcapsule suspension, and the mixture was slowly stirred with a propeller at room temperature for 15 minutes. Next, the microcapsules are collected by centrifugation (about 1,500 rpm),
After washing twice with ml, 180 mg of D-mannitol
Was added and freeze-dried to obtain a freeze-dried microcapsule powder containing hGH. Furthermore, in order to remove the residual solvent, vacuum drying was performed at 46 ° C. for 72 hours to obtain microcapsules.

Reference Example 4 Evaluation of Pharmacological Efficacy of Human Growth Hormone-Containing Microcapsules The administration of the immunosuppressant tacrolimus (Prograf Injection, Fujisawa Pharmaceutical Co., Ltd.) to female SD rats pituitary at 4 weeks of age was performed to produce antibodies against hGH. 5 weeks after the administration of microcapsules at 6 weeks of age, body weight, body length and serum rat insulin-like growth factor I (rIGF-I)
The concentration was measured. Specifically, Prograf Injection 5mg
Was diluted with physiological saline, and three days before and immediately after administration of the microcapsules, administrations 4, 7, and 11 were performed.
At a dose of 50 μg / 0.2 ml / rat after 75 days and 75 days after administration at 18, 18, 21, 25, 28 and 32 days.
Subcutaneous injection at a dose of μg / 0.2 ml / rat. In addition, hormone replacement was also performed to make the hypophysectomized rats more physiologically normal. A mixture of sodium L-thyroxine pentahydrate and hydrocortisone succinate (both from Wako Pure Chemical Industries, Ltd.) (final concentrations of 1 μg and 50 μg / respectively)
0.2 ml / rat) 3 times a week, ie, 3 days before and immediately after the administration of the microcapsules, 2, 4, 7, 9, 1
1, 14, 16, 18, 21, 23, 25, 28, 30
And subcutaneous injection 32 days later. 2 microcapsules
It was dispersed in a dispersion medium (5% mannitol, 0.5% sodium carboxymethylcellulose, 0.1% Tween 80) to a concentration of 4 mg hGH / ml, and its 0.5 m
was administered subcutaneously to the back of the rat under ether anesthesia. The dose was 12 mg as hGH. After the administration of the microcapsules, the body weight and body length of the rats were measured over time until 35 days later. Blood was collected over time from the tail vein, and serum was collected. The serum rIGF-I concentration was determined by radioimmunoassay (DSL-2900, Diagnostic Systems Laboratories, I
nc.).

Reference Example 5 2.0 g of candesartan, zinc oxide (manufactured by Hakusui Chemical Industry)
0.37 g and lactic acid-glycolic acid copolymer (lactic acid /
Glycolic acid 75/25 (mol%), weight average molecular weight 8,700, 3.6 g) (manufactured by Wako Pure Chemical Industries, Ltd.) was added to a mixture of 12.75 ml of dichloromethane, 2.25 ml of methanol and 0.136 ml of acetic acid, and the mixture was added at room temperature. Shake overnight to obtain a homogeneous solution. This solution was poured into 800 ml of a 0.1% by weight aqueous solution of polyvinyl alcohol containing 20 mM zinc acetate which had been adjusted to 18 ° C. in advance, and an O / W emulsion was prepared at 7,000 rpm using a turbine-type homomixer. The O / W emulsion was stirred at room temperature for 3 hours to evaporate dichloromethane, methanol and acetic acid, and solidify the oil phase.
Collected at 000 rpm. After dispersing this in distilled water again,
Further, centrifugation was performed to wash free drugs and the like. The collected microcapsules were redispersed by adding distilled water in which 0.8 g of mannitol was dissolved, and then freeze-dried to obtain a powder. The encapsulation rate of candesartan in the microcapsules was 90.9%, and the content of candesartan in the microcapsules / mannitol powder was 26.5%.

Reference Example 6 One batch was processed at the following throughput. 1. Candesartan
0 g, zinc oxide (manufactured by Hakusui Chemical Industry) 0.37 g, and lactic acid-glycolic acid copolymer (lactic acid / glycolic acid 75 /
25 (mol%), weight average molecular weight 8,700, manufactured by Wako Pure Chemical Industries, Ltd.) (3.6 g) was added to a mixture of dichloromethane (12.75 ml), methanol (2.25 ml) and acetic acid (0.136 ml), and the mixture was shaken overnight at room temperature with stirring. This gave a homogeneous solution. This solution was adjusted to 18 ° C. in advance and added with 10% zinc acetate-added 0.1% by weight aqueous polyvinyl alcohol solution 8
100 ml, using a turbine-type homomixer,
An O / W emulsion was obtained at 7,000 rpm. This O
The / W emulsion was stirred at room temperature for 3 hours to evaporate dichloromethane, methanol and acetic acid to solidify the oil phase, and then collected at 3,000 rpm using a centrifuge. This was dispersed again in distilled water and then centrifuged to wash free drugs and the like. After the above operation was performed in two batches, two batches of microcapsules were mixed, distilled water in which 1.6 g of mannitol was dissolved was added, and redispersion was performed.
Lyophilized to give a powder. The encapsulation rate of candesartan in the microcapsules was 90.7%, and the content of candesartan in the microcapsules / mannitol powder was 26.4%.

Example 1 Using 0.3 g of each of the freeze-dried microcapsules containing hGH obtained in Reference Example 1, a solvent was removed under the following four conditions. The microcapsule powder was transferred to an extraction container (capacity: 10 ml) of a supercritical fluid extraction device manufactured by JASCO Corporation, the device was sealed, and then heated to a predetermined temperature in a thermostat. Next, pressurize carbon dioxide to cylinder pressure (about 6-7MP).
In a), the liquid is sent to a heat exchanger via a liquid sending pump (SCF-Get), and is heated to a predetermined temperature, and a fully automatic pressure regulating valve (S
CF-Bpg) controls the pressure applied to the entire system,
It was converted to a high pressure gas state at a predetermined pressure. Next, the high-pressure carbon dioxide gas was blown into the extraction container, and the solvent was removed under the following four conditions. (1) Pressure 2MPa, temperature 15 ° C, extraction time 15 minutes. (2) Pressure 2 MPa, temperature 15 ° C., extraction time 30 minutes. (3) Pressure 2 MPa, temperature 15 ° C., extraction time 45 minutes. (4) Pressure 2 MPa, temperature 15 ° C., extraction time 60 minutes.

Example 2 Using 0.3 g each of the freeze-dried microcapsule powder containing hGH obtained in Reference Example 2, the solvent was removed under the following four conditions. The microcapsule powder was transferred to an extraction container (capacity: 10 ml) of a supercritical fluid extraction device manufactured by JASCO Corporation, the device was sealed, and then heated to a predetermined temperature in a thermostat. Next, pressurize carbon dioxide to cylinder pressure (about 6-7MP).
In a), the liquid is sent to a heat exchanger via a liquid sending pump (SCF-Get), and is heated to a predetermined temperature, and a fully automatic pressure regulating valve (S
CF-Bpg) controls the pressure applied to the entire system,
It was converted to a high pressure gas state at a predetermined pressure. Next, the high-pressure carbon dioxide gas was blown into the extraction container, and the solvent was removed under the following four conditions. (1) Pressure 2MPa, temperature 15 ° C, extraction time 30 minutes. (2) Pressure 2 MPa, temperature 15 ° C., extraction time 60 minutes. (3) Pressure 2 MPa, temperature 15 ° C., extraction time 180 minutes. (4) Pressure 1 MPa, temperature 15 ° C., extraction time 720 minutes.

Example 3 The lyophilized microcapsule powder containing candesartan obtained in Reference Example 5 was used in an amount of 0.3 g each to obtain the following 1
The solvent was removed under eight conditions. The microcapsule powder was transferred to an extraction container (capacity: 10 ml) of a supercritical fluid extraction device manufactured by JASCO Corporation, the device was sealed, and then heated to a predetermined temperature in a thermostat. Next, carbon dioxide is sent to the heat exchanger via a liquid sending pump (SCF-Get) at a cylinder pressure (about 6 to 7 MPa), and is heated to a predetermined temperature, and then is fully automatic pressure regulating valve (SCF-Bpg). Thus, the pressure applied to the entire system was controlled to convert the system into a high-pressure gas state at a predetermined pressure. Next, the high-pressure carbon dioxide gas was blown into the extraction container, and the solvent was removed under the following 18 conditions. Pressure 2.0MPa, temperature 15 ° C, extraction time 30 minutes. Pressure 2.0MPa, temperature 15 ° C, extraction time 60 minutes. Pressure 2.0MPa, temperature 15 ° C, extraction time 120 minutes. Pressure 2.0MPa, temperature 15 ° C, extraction time 180 minutes. Pressure 2.5MPa, temperature 15 ° C, extraction time 30 minutes. Pressure 2.5MPa, temperature 15 ° C, extraction time 60 minutes. Pressure 2.5MPa, temperature 15 ° C, extraction time 120 minutes. Pressure 2.5MPa, temperature 15 ° C, extraction time 180 minutes. The pressure was 3.0 MPa, the temperature was 15 ° C., and the extraction time was 15 minutes. (10) Pressure 3.0 MPa, temperature 15 ° C., extraction time 30
Minutes. (11) Pressure 3.0 MPa, temperature 15 ° C., extraction time 60
Minutes. (12) Pressure 3.0 MPa, temperature 15 ° C., extraction time 12
0 minutes. (13) Pressure 3.0 MPa, temperature 15 ° C., extraction time 18
0 minutes. (14) Pressure 3.5 MPa, temperature 15 ° C., extraction time 30
Minutes. (15) Pressure 3.5 MPa, temperature 15 ° C., extraction time 60
Minutes. (16) Pressure 3.5 MPa, temperature 15 ° C., extraction time 12
0 minutes. (17) Pressure 3.5 MPa, temperature 15 ° C., extraction time 18
0 minutes. (18) Pressure 4.0 MPa, temperature 15 ° C., extraction time 30
Minutes.

Example 4 The freeze-dried microcapsule powder containing candesartan obtained in Reference Example 6 was subjected to a solvent removal treatment under the following three conditions. The microcapsule powder was transferred to an extraction container (capacity: 10 ml) of a supercritical fluid extraction device manufactured by JASCO Corporation, the device was sealed, and then heated to a predetermined temperature in a thermostat. Next, carbon dioxide is sent to the heat exchanger via a liquid sending pump (SCF-Get) at a cylinder pressure (about 6 to 7 MPa), and is heated to a predetermined temperature, and is then fully heated.
-Bpg) to control the pressure applied to the entire system to convert it into a high-pressure gas state at a predetermined pressure. Next, the high-pressure carbon dioxide gas was blown into the extraction container, and the solvent was removed under the following three conditions. (1) Pressure 3.0 MPa, temperature 15 ° C., extraction time 60 minutes, charged amount of microcapsules 0.3 g. (2) Pressure 3.0 MPa, temperature 15 ° C., extraction time 60 minutes, charged amount of microcapsules 2 g. (3) Pressure 3.0 MPa, temperature 15 ° C., extraction time 60 minutes, charged amount of microcapsules 5 g.

Test Example 1 With respect to the hGH-containing microcapsules and the untreated freeze-dried microcapsules obtained in Examples 1 (1) to (4), the residual dichloromethane (DCM) content and the hGH content in the microcapsules were as follows. Was measured by the following method. (1) Residual dichloromethane (DCM) amount About 100 mg of microcapsules were precisely weighed and dissolved in dimethyl sulfoxide to make exactly 5 ml, which was used as a sample solution. Separately, about 1 g of dichloromethane was precisely weighed, and dimethyl sulfoxide was added to make exactly 20 ml. This solution was diluted exactly 10000-fold with dimethyl sulfoxide to obtain a standard solution. A test was performed on 1 μl of the sample solution and the standard solution by gas chromatography under the following conditions, and the peak area of dichloromethane in each solution was measured by an automatic integration method to calculate the amount of dichloromethane. Detector: Hydrogen flame ionization detector Column: OVI-G43 Film thickness 3μm, 0.53mm id ×
30m (Supelco) Inlet temperature: 140 ° C Detector temperature: 260 ° C Column temperature: 40 ° C (10 min hold) → 240 ° C
(35 ° C./min)→240° C. (hold 20 min) → Cooling → 40 ° C. Carrier gas: helium Flow rate: 35 cm / sec (2) hGH content 10 mg of microcapsules are precisely weighed into a 5 ml volumetric flask, and 1.75 ml. After addition of acetonitrile, it was subjected to sonication. 3m in the obtained acetonitrile solution
of 150 mM phosphate buffer (pH 8.0) was added, and after sonication, 150 mM phosphate buffer (pH 8.0) was added.
8.0). 1 ml of this is 1500
Centrifuge at 0 rpm for 10 minutes.
The solution was filtered through a μm membrane filter. Then this h
The GH extract was subjected to size exclusion high performance liquid chromatography under the following conditions to measure the hGH content. Column: TSK-gel G2000SWXL, 7.8 mm id ×
300 mm (Tosoh) Mobile phase: 0.05 mol / l Ammonium hydrogen carbonate solution Flow rate: 0.6 ml / min The results are shown in Table 1.

[0076]

[Table 1] As is clear from the results in Table 1, the amount of residual dichloromethane was significantly reduced in the microcapsules treated with carbon dioxide in a high-pressure gas state as compared with the untreated microcapsules. Further, it was confirmed that the hGH content in the microcapsules did not decrease by the carbon dioxide treatment in a high-pressure gas state.

Test Example 2 For the hGH-containing microcapsules and the untreated freeze-dried microcapsules obtained in Examples 1 (1) to (4), the amounts of the hGH polymer and the hGH-related protein in the microcapsules were determined. It was measured by the following method. (1) 10 mg of hGH polymer microcapsules were accurately weighed, 2.5 ml of acetonitrile was added, and the sample was dispersed by ultrasonic irradiation. Subsequently, ultrasonic irradiation was performed for about 2 minutes, followed by centrifugation at 3000 rpm for 10 minutes. did. The supernatant was removed, 2.5 ml of acetonitrile was added to the residue, the residue was dispersed by ultrasonic irradiation, followed by ultrasonic irradiation for about 2 minutes, and then centrifuged at 3000 rpm for 10 minutes. The supernatant was removed and the residue was dried in a desiccator under reduced pressure.
To this, 1.25 ml of a diluent (phosphate buffer (pH 8.0) / acetonitrile mixed solution (13: 7)) was added, and the sample was dispersed by ultrasonic irradiation. Subsequently, ultrasonic irradiation was performed for about 2 minutes. The solution was filtered through a 0.5 μm membrane filter, and the filtrate was used as a sample solution. Separately, 0.2 ml of hGH standard was added to 0.2 ml of diluent. This liquid 0.2
ml was added to 4.8 ml of the diluent to give a standard solution. The measurement was carried out on 50 μl of the sample solution and the standard solution by liquid chromatography under the following conditions. HG of sample solution
The peak area of the peak eluted earlier than the retention time of H and the hGH peak area of the standard solution were measured by an automatic integration method, and the content of the polymer was determined. At the same time, 50 μl of the diluent was injected, and the peak detected from the blank was subtracted from the calculation. Detector: UV absorption spectrophotometer (measuring wavelength: 214 nm) Column: TSK-gel G2000SWXL, 7.8 mm id x 300 mm
(Tosoh) Column temperature: constant temperature around 25 ° C Mobile phase: 0.05 mol / l ammonium bicarbonate solution Flow rate: 0.6 ml / min

(2) hGH related protein Microcapsules 40 mg were precisely weighed, 2 ml of acetonitrile was added, and the sample was dispersed by ultrasonic irradiation.
Subsequently, ultrasonic irradiation was performed for about 2 minutes. Next, 3 ml of a phosphate buffer (pH 8.0) was added, and the mixture was irradiated with ultrasonic waves for about 2 minutes with occasional shaking, and then centrifuged at 4 ° C., 3500 times / minute for 10 minutes. The supernatant was filtered through a membrane filter having a pore size of 0.5 μm, and the filtrate was used as a sample solution. Separately, 0.1 ml of the hGH standard was diluted with a diluent (phosphate buffer (pH 8.0) / acetonitrile mixture (13:
7)) In addition to 3.9 ml, a standard solution was prepared. 20 μl of the sample solution and the standard solution were measured by liquid chromatography under the following conditions. The individual peak areas other than hGH in the sample solution and the hGH peak areas in the standard solution were measured by the automatic integration method, and the content of the related proteins was determined. At the same time, 20 μl of the diluted solution was injected, and the peak detected from the blank was subtracted from the calculation. Detector: UV absorption spectrophotometer (measuring wavelength: 220 nm) Column: PROTEIN C4, 4.6 mm id × 250 mm (VYDA
C) Column temperature: constant temperature around 45 ° C Mobile phase: (A) 2-amino-2-hydroxymethyl-1,3-propanediol buffer solution (pH 7.5) / 1-propanol mixed solution (1
9: 6) (B) 2-Amino-2-hydroxymethyl-1,3-propanediol buffer solution (pH 7.5) / 1-propanol mixed solution (17: 8) Solution (A) and solution (B) Flow at a ratio of 1: 1). Flow rate: 0.5 ml / min The results are shown in Table 2.

[0079]

[Table 2] As is clear from the results in Table 2, the amount of the hGH polymer and the hGH analogous protein was not increased in the microcapsules treated with carbon dioxide in a high-pressure gas state as compared with the untreated microcapsules.

Test Example 3 With respect to the hGH-containing microcapsules and the untreated freeze-dried microcapsules obtained in Examples 1 (1) to (4), the average particle size of the microcapsules and
The in vivo initial release rate was measured by the following method. (1) Average particle diameter of microcapsules The average particle diameter of microcapsules is measured using a particle size distribution analyzer (Mu
ltisizerII, Coulter Electronics Ltd., Beds, UK). (2) Initial release rate in vivo Rats were treated with tacrolimus for immunosuppression. Prograf Injection 5 mg (Fujisawa Pharmaceutical) was diluted with physiological saline, and 0.4 mg /
0.2 ml / rat, immediately after microcapsule administration, 0.2 mg / 0.2 ml / rat 4, 7, and 11 days after administration, 0.3 mg / 0.2 ml after 14, 18, 21, 25, 28, and 32 days after administration By subcutaneous administration at a dose of / rat, antibody production against hGH could be suppressed, and it became possible to measure the concentration of hGH in rat serum over 5 weeks after administration. Microcapsules are 16mg hG
H / ml (5% mannitol, 0.1% mannitol).
5% carboxymethylcellulose, 0.1% Tween
80). 0.75 ml of the obtained dispersion was subcutaneously administered to the back of the rat under ether anesthesia. The dose is hG
H was 12 mg. After administration of the microcapsules, blood was collected over time from the tail vein, and the serum was collected. HG in serum
The measurement of H concentration is performed by immunoradiometric assay (Ab
(Beads HGH, Eiken Chemical Co., Ltd.). The hGH solution was subcutaneously administered to immunosuppressed rats at doses of 5, 10 and 20 mg / kg, blood was collected over time, and serum hGH was
The concentration was measured. AUC was calculated by the trapezoidal method. From the AUC up to 24 hours after the microcapsule administration, calculate the hGH dose corresponding to that in the case of hGH solution subcutaneous administration,
The initial release rate was calculated by dividing by the microcapsule dose of 12 mg. Table 3 shows the results.

[0081]

[Table 3] As is clear from the results in Table 3, it was confirmed that the average particle size of the microcapsules did not change due to the carbon dioxide treatment in a high-pressure gas state, and aggregation did not occur. In addition, the initial release rate of microcapsules treated with carbon dioxide in a high-pressure gas state was significantly reduced as compared with untreated microcapsules.

Test Example 4 For the hGH-containing microcapsules and the untreated freeze-dried microcapsules obtained in Examples 2 (1) to (4), the residual dichloromethane (DCM) content and the hGH content in the microcapsules were as follows: Was measured by the following method. (1) Residual dichloromethane (DCM) amount About 100 mg of microcapsules were precisely weighed and dissolved in dimethyl sulfoxide to make exactly 5 ml, which was used as a sample solution. Separately, about 1 g of dichloromethane was precisely weighed, and dimethyl sulfoxide was added to make exactly 20 ml. This solution was diluted exactly 10000-fold with dimethyl sulfoxide to obtain a standard solution. A test was performed on 1 μl of the sample solution and the standard solution by gas chromatography under the following conditions, and the peak area of dichloromethane in each solution was measured by an automatic integration method to calculate the amount of dichloromethane. Detector: Hydrogen flame ionization detector Column: OVI-G43 Film thickness 3μm, 0.53mm id ×
30m (Supelco) Inlet temperature: 140 ° C Detector temperature: 260 ° C Column temperature: 40 ° C (10 min hold) → 240 ° C
(35 ° C./min)→240° C. (hold 20 min) → Cooling → 40 ° C. Carrier gas: helium Flow rate: 35 cm / sec (2) hGH content 20 mg of microcapsules are precisely weighed into a 5 ml volumetric flask, and 1.75 ml. After addition of acetonitrile, it was subjected to sonication. 3m in the obtained acetonitrile solution
of 150 mM phosphate buffer (pH 8.0) was added, and after sonication, 150 mM phosphate buffer (pH 8.0) was added.
8.0). 1 ml of this is 1500
Centrifuge at 0 rpm for 10 minutes.
The solution was filtered through a μm membrane filter. Then this h
The GH extract was subjected to size exclusion high performance liquid chromatography under the following conditions to measure the hGH content. Column: TSK-gel G2000SWXL, 7.8mm id × 300mm
(Tosoh) Mobile phase: 0.05 mol / l Ammonium hydrogen carbonate solution Flow rate: 0.6 ml / min The results are shown in Table 4.

[0083]

[Table 4] As is clear from the results in Table 4, the amount of residual dichloromethane was significantly reduced in the microcapsules treated with carbon dioxide in a high-pressure gas state as compared with the untreated microcapsules. Further, it was confirmed that the hGH content in the microcapsules did not decrease by the carbon dioxide treatment in a high-pressure gas state.

Test Example 5 For the hGH-containing microcapsules and the untreated freeze-dried microcapsules obtained in Examples 2 (1) to (4), the amounts of the hGH polymer and the hGH-related protein in the microcapsules were determined. It was measured by the following method. (1) hGH polymer 10 mg of microcapsules were precisely weighed, 2.5 ml of acetonitrile was added, and the sample was dispersed by ultrasonic irradiation. Subsequently, ultrasonic irradiation was performed for about 2 minutes, followed by centrifugation at 3000 rpm for 10 minutes. did. The supernatant was removed, 2.5 ml of acetonitrile was added to the residue, the residue was dispersed by ultrasonic irradiation, followed by ultrasonic irradiation for about 2 minutes, and then centrifuged at 3000 rpm for 10 minutes. The supernatant was removed and the residue was dried in a desiccator under reduced pressure.
To this, 1.25 ml of a diluent (phosphate buffer (pH 8.0) / acetonitrile mixed solution (13: 7)) was added, and the sample was dispersed by ultrasonic irradiation. Subsequently, ultrasonic irradiation was performed for about 2 minutes. The solution was filtered through a 0.5 μm membrane filter, and the filtrate was used as a sample solution. Separately, 0.2 ml of hGH standard was added to 0.2 ml of diluent. This liquid 0.2
ml was added to 4.8 ml of the diluent to give a standard solution. The measurement was carried out on 50 μl of the sample solution and the standard solution by liquid chromatography under the following conditions. HG of sample solution
The peak area of the peak eluted earlier than the retention time of H and the hGH peak area of the standard solution were measured by an automatic integration method, and the content of the polymer was determined. 50μl of diluent at the same time
And peaks detected from the blank were subtracted from the calculation. Detector: UV absorption spectrophotometer (measuring wavelength: 214 nm) Column: TSK-gel G2000SWXL, 7.8 mm id x 300 mm
(Tosoh) Column temperature: constant temperature around 25 ° C Mobile phase: 0.05 mol / l ammonium bicarbonate solution Flow rate: 0.6 ml / min (2) hGH related protein Microcapsules 40 mg are precisely weighed, and acetonitrile 2 ml is added. And disperse the sample by ultrasonic irradiation,
Subsequently, ultrasonic irradiation was performed for about 2 minutes. Next, 3 ml of a phosphate buffer (pH 8.0) was added, and the mixture was irradiated with ultrasonic waves for about 2 minutes with occasional shaking, and then centrifuged at 4 ° C., 3500 times / minute for 10 minutes. The supernatant was filtered through a membrane filter having a pore size of 0.5 μm, and the filtrate was used as a sample solution. Separately, 0.1 ml of the hGH standard was diluted with a diluent (phosphate buffer (pH 8.0) / acetonitrile mixture (13:
7)) In addition to 3.9 ml, a standard solution was prepared. 20 μl of the sample solution and the standard solution were measured by liquid chromatography under the following conditions. The individual peak areas other than hGH in the sample solution and the hGH peak areas in the standard solution were measured by the automatic integration method, and the content of the related proteins was determined. At the same time, 20 μl of the diluted solution was injected, and the peak detected from the blank was subtracted from the calculation. Detector: UV absorption spectrophotometer (measuring wavelength: 220 nm) Column: PROTEIN C4, 4.6 mm id × 250 mm (VYDA
C) Column temperature: constant temperature around 45 ° C Mobile phase: (A) 2-amino-2-hydroxymethyl-1,3-propanediol buffer solution (pH 7.5) / 1-propanol mixed solution (19: 6) ( B) 2-Amino-2-hydroxymethyl-1,3-propanediol buffer solution (pH 7.5) / 1-propanol mixed solution (17: 8) Solution (A) and solution (B) were mixed with (1: 1) Pour at a rate. Flow rate: 0.5 ml / min The results are shown in Table 5.

[0085]

[Table 5] As is clear from the results in Table 5, the amounts of the hGH polymer and the hGH analogous protein were not increased in the microcapsules treated with carbon dioxide in a high-pressure gas state as compared with the untreated microcapsules.

Test Example 6 For the hGH-containing microcapsules and the untreated freeze-dried microcapsules obtained in Examples 2 (1) to (4), the average particle size of the microcapsules and
The in vivo initial release rate was measured by the following method. (1) Average particle size of microcapsules The average particle size of microcapsules is measured using a particle size distribution analyzer (Mu
ltisizerII, Coulter Electronics Ltd., Beds, UK). (2) Initial release rate in vivo Rats were treated with tacrolimus for immunosuppression. Prograf Injection 5 mg (Fujisawa Pharmaceutical) was diluted with physiological saline, and 0.4 mg /
0.2 ml / rat, immediately after administration of the microcapsules, and 0.2 mg / 0,0,4, 7, 11, 14 and 18 days after administration.
It was administered subcutaneously at a dose of 2 ml / rat. The microcapsules were dispersed in a dispersion medium (5%
Mannitol, 0.5% carboxymethylcellulose,
0.1% Tween 80). 0.75 ml of the obtained dispersion was subcutaneously administered to the back of the rat under ether anesthesia. The dose was 6 mg as hGH. After administration of the microcapsules, blood was collected over time from the tail vein, and the serum was collected. The serum hGH concentration was measured by an immunoradiometric assay (Ab beads HGH, Eiken Chemical). The hGH solution was given to immunosuppressed rats at 5, 10 and 20
The solution was subcutaneously administered at a dose of mg / kg, blood was collected over time, and the serum hGH concentration was measured. AUC was calculated by the trapezoidal method. AUC up to 24 hours after microcapsule administration
To hGH equivalent to subcutaneous administration of hGH solution
The dose was calculated and the initial release rate was calculated by dividing by the microcapsule dose of 6 mg. Table 6 shows the results.

[0087]

[Table 6] As is clear from the results in Table 6, it was confirmed that the average particle size of the microcapsules did not change by carbon dioxide treatment in a high-pressure gas state, and aggregation did not occur. In addition, the initial release rate of microcapsules treated with carbon dioxide in a high-pressure gas state was significantly reduced as compared with untreated microcapsules.

Test Example 7 Regarding the candesartan-containing microcapsules obtained in Examples 3 (1) to (18) and the untreated freeze-dried microcapsules, the residual dichloromethane (DCM) content and candesartan content in the microcapsules were as follows. Was measured by the following method. Residual dichloromethane (DCM) amount About 100 mg of microcapsules were precisely weighed and dissolved in dimethyl sulfoxide to make exactly 5 ml, which was used as a sample solution. Separately, about 1 g of dichloromethane was precisely weighed, and dimethyl sulfoxide was added to make exactly 20 ml. This solution was diluted exactly 10000-fold with dimethyl sulfoxide to obtain a standard solution. A test was performed on 1 μl of the sample solution and the standard solution by gas chromatography under the following conditions, and the peak area of dichloromethane in each solution was measured by an automatic integration method to calculate the amount of dichloromethane. Detector: Hydrogen flame ionization detector Column: OVI-G43 Film thickness 3μm, 0.53mm id × 30
m (Supelco) Inlet temperature: 140 ° C Detector temperature: 260 ° C Column temperature: 40 ° C (10 min hold) → 260 ° C (35 ° C / mi)
n) (10 min hold) Carrier gas: helium Flow rate: 35 cm / sec Candesartan content 5 to 10 mg of microcapsules are accurately weighed into a centrifuge tube,
After adding 30 ml of the HPLC mobile phase, the mixture was shaken and stirred for 1 hour. Subsequently, it was centrifuged at 2950 rpm for 10 minutes,
The supernatant was filtered with a membrane filter having a pore size of 0.5 μm. Next, this candesartan extract was subjected to reversed-phase high-performance liquid chromatography under the following conditions to measure the content of candesartan. Column: inertsil ODS-3 (4.6 mm × 150 mm, GL science) Mobile phase: 0.1 M KH ₂ PO ₄ / AcCN / MeOH / AcOH = 50/35/15/1 (v /
v) Flow rate: 1 ml / min Detection: UV wavelength 254 nm The results are shown in Table 7.

[0089]

[Table 7] As is clear from the results in Table 7, the amount of residual dichloromethane was significantly reduced in the microcapsules treated with carbon dioxide in a high-pressure gas state as compared with the untreated microcapsules. It was also confirmed that the content of candesartan in the microcapsules did not decrease by carbon dioxide treatment in a high-pressure gas state.

Test Example 8 For the microcapsules containing candesartan and the untreated freeze-dried microcapsules obtained in Examples 4 (1) to (3), the residual dichloromethane (DCM) content and the content of candesartan in the microcapsules were tested. It was measured in the same manner as in Example 7. Table 8 shows the results.

[0091]

[Table 8] As is clear from the results in Table 8, the amount of residual dichloromethane was significantly reduced in the microcapsules treated with carbon dioxide in a high-pressure gas state, as compared with the untreated microcapsules. It was also confirmed that the content of candesartan in the microcapsules did not decrease by carbon dioxide treatment in a high-pressure gas state.

[0092]

According to the present invention, in a method for producing a sustained-release preparation, a solid containing a physiologically active substance and a biodegradable polymer is formed, and the solid is contacted with a high-pressure gas to administer the solid. Immediately after the initial release of an excessive amount of the physiologically active substance is drastically suppressed, a fixed amount of the physiologically active substance is released over a long period immediately after administration, and the bioactive substance has very little alteration and residual organic solvent. Clinically, a sustained release preparation having extremely excellent properties can be obtained. In addition, the processing time required for removing the solvent was significantly reduced by changing the method for removing the solvent.

[Brief description of the drawings]

FIG. 1 is a schematic view showing a solvent removing apparatus using carbon dioxide in a high-pressure gas state.

[Explanation of symbols]

1: liquefied carbon dioxide cylinder, 2: CO2 feed pump, 3: heat exchanger, 4: extraction vessel, 5: constant temperature bath, 6: detector, 7: automatic pressure regulating valve, 8: recovery vessel

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI theme coat ゛ (Reference) A61K 31/522 A61K 31/522 38/00 45/00 38/27 47/30 45/00 A61P 1/00 47/30 1/14 A61P 1/00 1/16 1/14 3/00 1/16 3/04 3/00 3/06 3/04 3/10 3/06 5/08 3/10 7/00 5 / 08 7/02 7/00 7/04 7/02 7/06 7/04 9/04 7/06 9/10 9/04 101 9/10 13/12 101 17/02 13/12 19/00 17 / 02 19/10 19/00 25/00 19/10 25/16 25/00 25/28 25/16 27/06 25/28 35/00 27/06 37/04 35/00 43/00 37/04 A61K 37/02 43/00 37/36 F-term (reference) 4C076 AA67 BB11 CC01 CC04 CC11 CC14 CC17 CC21 CC26 CC27 CC30 CC34 EE24 FF01 FF31 FF65 FF68 GG30 4C084 AA02 AA03 AA17 AA27 BA01 BA08 BA23 BA05 CA53 MA62 DB03 MA05 NA10 NA12 NA13 ZA052 ZA062 ZA072 ZA082 ZA122 ZA362 ZA392 ZA422 ZA532 ZA542 ZA622 ZA632 ZA682 ZA832 ZA942 ZA962 ZB132 ZB212 ZB 262 ZB352 ZC032 ZC332 ZC352 4C086 AA02 BC38 BC39 BC62 CB09 GA02 GA04 GA07 MA01 MA02 MA04 MA05 MA38 MA66 NA03 NA10 NA12 NA13 ZA05 ZA06 ZA07 ZA08 ZA12 ZA36 ZA39 ZA42 ZA53 ZA54 ZA36 ZA63 ZAZ ZA69

Claims (31)

[Claims] 1. A method for producing a preparation containing a physiologically active substance, comprising forming a solid containing a physiologically active substance and a polymer, and contacting the solid with a high-pressure gas. 2. The method according to claim 1, wherein the physiologically active substance is a physiologically active substance which is unstable to heat or a solvent. 3. The method according to claim 1, wherein the bioactive substance is a bioactive peptide having a molecular weight of about 2,000 to about 500,000. 4. The method according to claim 1, wherein the bioactive substance is a bioactive peptide having a molecular weight of about 5,000 to about 500,000. 5. The method according to claim 4, wherein the physiologically active peptide is human growth hormone. 6. The method according to claim 1, wherein the physiologically active substance is a non-peptidic compound. 7. The method according to claim 6, wherein the non-peptidic compound is a compound having an oxygen atom in the molecule. 8. The method according to claim 6, wherein the non-peptidic compound is a compound having an ether bond or a carbonyl group. 9. The non-peptidic compound of the formula (I) (Wherein, R ¹ represents a group capable of forming an anion or a group capable of changing to an anion; X represents that a phenylene group and a phenyl group are bonded directly or via a spacer having an atomic chain of 2 or less; represents an integer of 1 or 2, ring a denotes a benzene ring optionally further have a substituent, R ² represents a group capable of converting thereinto or a group capable of forming an anion, R ³
Represents a hydrocarbon residue which may be bonded via a hetero atom and may have a substituent) or a salt thereof. 10. The method according to claim 6, wherein the non-peptidic compound is losartan, eprosartan, candesartan cilexetil, candesartan, valsartan, telmisartan, irbesartan, tasosartan or olmesartan. 11. The method according to claim 6, wherein the non-peptidic compound is candesartan. 12. The method according to claim 1, wherein the polymer is biodegradable. 13. The method according to claim 12, wherein the biodegradable polymer is a homo- or copolymer of α-hydroxycarboxylic acids, or a mixture thereof. 14. The production according to claim 13, wherein the biodegradable polymer is a lactic acid / glycolic acid homopolymer or copolymer having a lactic acid / glycolic acid composition ratio of about 100/0 to about 40/60 mol%. Law. 15. The method according to claim 13, wherein the biodegradable polymer is a lactic acid homopolymer. 16. The method according to claim 12, wherein the biodegradable polymer has a weight average molecular weight of about 3,000 to about 50,000. 17. The method of claim 1 wherein the solid is contacted with a high pressure gas within a temperature range of about +20 to about -60 ° C., based on the glass transition temperature of the polymer. 18. The method of claim 17, wherein the solid is contacted with a propellant within a temperature range of about 0 to about -40 ° C., based on the glass transition temperature of the polymer. 19. The method of claim 1, wherein the time of contacting the solid with the high pressure gas is from about 5 minutes to about 48 hours. 20. The method of claim 19, wherein the time of contacting the solid with the high pressure gas is from about 10 minutes to about 12 hours. 21. The method according to claim 1, wherein the high-pressure gas is a substance inert to a physiologically active substance and a polymer. 22. The high-pressure gas is carbon dioxide.
1. The production method according to 1. 23. The method according to claim 1, wherein the pressure of the high-pressure gas is about 1 to about 7 MPa. 24. The method according to claim 23, wherein the pressure of the high-pressure gas is about 1 to about 4 MPa. 25. The method according to claim 1, wherein the preparation is a sustained release microcapsule. 26. The method according to claim 25, wherein the sustained-release microcapsules are obtained by an underwater drying method. 27. A preparation obtained by the production method according to claim 1. 28. Sustained-release microcapsules obtained by the production method according to claim 25. 29. An injection comprising the sustained-release microcapsule according to claim 28. 30. A method for suppressing the initial release of a physiologically active substance, comprising forming a solid containing a physiologically active substance and a polymer, and contacting the solid with a high-pressure gas. 31. A method for suppressing deterioration of a physiologically active substance, comprising forming a solid containing a physiologically active substance and a polymer, and contacting the solid with a high-pressure gas.