JP2002068982A - Sustained release composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a composition (1) increased in the content of a non-peptidic physiologically active substance and capable of exerting sure pharmacological effect through controlling or accelerating the release of the non-peptidic physiologically active substance, and (2) when the non-peptidic physiologically active substance had subcutaneous irritancy, hopeful of the action of compensating the irritancy by terminal intense acidic groups, and (3) having a high glass transition point and high stability. SOLUTION: This composition comprises a non-peptidic physiologically active substance and a biodegradable polymer or a salt thereof having at least two terminal carboxy groups.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a composition comprising a non-peptidic physiologically active substance and a biodegradable polymer having two or more carboxyl groups at a terminal or a salt thereof.

[0002]

2. Description of the Related Art The following method has been reported as a preparation for sustained release of a drug using a biodegradable polymer. 1) Pharmaceutical composition comprising a non-peptidic bone formation promoting substance and a biodegradable polymer (JP-A-9-2
63545). 2) Sustained-release preparations containing a compound having an angiotensin II antagonistic activity and a biodegradable polymer (JP-A-11-315034). 3) one ionically linked to a biologically active polypeptide comprising at least one effective ionogenic amine
A composition comprising a polyester containing the free COOH group described above, wherein at least 50% by weight of the polypeptide present in the composition is ionically bonded to the polyester [WO 94/15587 ( JP-T 8-505395)].

[0003]

Although a system for controlling the release of a non-peptidic physiologically active substance has been studied, it has not been possible to control the release rate, and in particular an effective means capable of accelerating the release. Has not yet been obtained.

[0004]

Means for Solving the Problems The present inventors have intensively studied to solve the above-mentioned problems, and have found that a non-peptidic bioactive substance and a biodegradable polymer having two or more carboxyl groups at a terminal thereof. Or the first time a composition comprising or blended with a salt thereof is produced, the release of the non-peptidic bioactive substance can be unexpectedly adjusted, especially when the non-peptidic bioactive substance is poorly water-soluble. Was found to accelerate. The present invention has been completed based on these findings. That is, the present invention provides: (1) a composition comprising a non-peptidic bioactive substance and a biodegradable polymer having two or more carboxyl groups at a terminal or a salt thereof; (2) a non-peptidic bioactivity A composition comprising a substance and a biodegradable polymer having two or more carboxyl groups at the terminal or a salt thereof; (3) a glass transition point (Tg) of the composition having two or more carboxyl groups at the terminal; The composition according to the above (2), wherein the temperature is higher than the glass transition point of the biodegradable polymer by at least about 10 ° C .; (4) The biodegradable polymer having two or more carboxyl groups at the terminal Is a polymer having an α, α-dicarboxyl group or an α, β, β′-tricarboxyl group at the terminal; (5) a composition according to the above (1) or (2); The biodegradable polymer having a ruboxyl group is a poly-α having an α, α-dicarboxyl group or an α, β, β′-tricarboxyl group at a terminal.
(1) or (2) above, which is a hydroxycarboxylic acid.
(6) The composition according to the above (5), wherein the poly-α-hydroxycarboxylic acid is linear; (7) a biodegradable polymer having two or more carboxyl groups at the terminal, Lactic acid having a terminal α, α-dicarboxyl group or α, β, β′-tricarboxyl group
(8) The composition according to the above (1) or (2), which is a glycolic acid polymer; (8) the biodegradable polymer having two or more carboxyl groups at its terminal, wherein the ω residue is tartronic acid or citric acid (9) the composition according to the above (1) or (2), which is a lactic acid-glycolic acid polymer; (9) the biodegradable polymer having two or more carboxyl groups at its terminal, wherein the ω residue is tartronic acid or citric acid (10) The composition according to (1) or (2), wherein the non-peptide physiologically active substance is poorly water-soluble. ) The composition according to (10), which is sustained-release; (12) the non-peptide physiologically active substance has a molecular weight of about 1,
(13) The composition according to (11), wherein the non-peptide physiologically active substance is a gonadotropin-releasing hormone agonist or antagonist; The active substance has the formula

Embedded image Wherein, X is a carbon atom or a nitrogen atom, - - - indicates a single bond or a double bond], or a salt thereof having a partial structure represented by [hereinafter, sometimes abbreviated as Compound (A) (15) a compound of the formula (11):

Embedded image Wherein R ¹ and R ² each represent a hydrogen atom, a hydroxy group, a C _1-4 alkoxy group, a C _1-4 alkoxy-carbonyl group or a C _1-4 alkyl group which may have a substituent. , R ³ represents a hydrogen atom, a halogen atom, a hydroxy group or a C _1-4 alkoxy group which may have a substituent, or two adjacent R ³ are linked to form a C _1-4 alkylenedioxy. R ⁴ represents a hydrogen atom or a C _1-4 alkyl group, and R ⁶ represents a C 5 optionally substituted group.
_1-4 alkyl group or formula

Embedded image (Wherein R ⁵ represents a hydrogen atom or R ⁴ and R ⁵ may be linked to form a heterocyclic ring), and n represents an integer of 0 to 5 Or a salt thereof [hereinafter may be abbreviated as compound (I)]; (16) the compound according to (14); N-methylaminomethyl) -1- (2,6-difluorobenzyl) -6- [4- (3-methoxyureido) phenyl]
-3-phenylthieno [2,3-d] pyrimidine-2,
4 (1H, 3H) -dione or a salt thereof (1)
(4) the composition according to (1), wherein the compound (A) has the formula

Embedded image[Wherein, R⁹Is optionally substituted C_1-7Alkyl group,
Optionally substituted C _3-7Cycloalkyl group, substituted
C that may be_1-6Alkoxyamino group or substituted
An optionally substituted hydroxyamino group is represented by R^TenIs replaced
May be C_1-7Alkyl group or substituted
Each represents a good phenyl group;⁹Is unsubstituted C_1-7A
When it is a alkyl group, R^TenIs the substituted C_1-7Alkyl
Represents a group or substituted phenyl]
Or a salt thereof [hereinafter may be abbreviated as compound (VIII).
(18) The compound according to the above (14), wherein the compound (A) is 3- (N-benzyl-N-meth-
(Tylaminomethyl) -4,7-dihydro-5-isobutyi
Ryl-7- (2,6-difluorobenzyl) -2- [4
-[(1-hydroxycyclopropyl) carbonylamido
No] phenyl] -4-oxothieno [2,3-b] pyri
(19) the composition according to (1) or (2), which is for injection;
(20) The above (1) or (2), which is a sustained release microcapsule.
Is a composition according to (2); (21) a non-peptide physiologically active substance and two or more
Biodegradable polymer having carboxyl group or its
And a method for producing a composition characterized by blending with a salt of
About.

[0005] The "non-peptidic physiologically active substance" used in the present invention, in particular, the "poorly water-soluble non-peptidic physiologically active substance" may be a free form or a salt. Examples of the “salt” include metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. Preferable examples of the metal salt include an alkali metal salt such as a sodium salt and a potassium salt; an alkaline earth metal salt such as a calcium salt, a magnesium salt and a barium salt; an aluminum salt. Preferred examples of the salt with an organic base include, for example, trimethylamine, triethylamine, pyridine,
Picoline, ethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, N, N
-Salts with dibenzylethylenediamine and the like. Preferred examples of the salt with an inorganic acid include, for example, salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Suitable examples of salts with organic acids include, for example, formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, p -Salts with toluenesulfonic acid and the like. Preferred examples of the salt with a basic amino acid include, for example, salts with arginine, lysine, ornithine, and preferred examples of the salt with an acidic amino acid include, for example, salts with aspartic acid, glutamic acid, and the like. Can be

The "non-peptidic physiologically active substance", particularly the "poorly water-soluble non-peptidic physiologically active substance" may be any pharmacologically useful substance, and is preferably a synthetic organic compound. Examples of the “synthetic organic compound” include a compound having a hydrophilic portion mainly having a tertiary amine and a highly hydrophobic portion such as a chain or cyclic alkyl or aromatic group, or a salt thereof. Specific examples include basic and amphiphilic drugs [CAD (cationic amphiphilic d
rug); Pharmacological Reviews (Pharmacologi)
cal Reviews), Vol. 42, No. 4, pp. 327-354,
1990 etc.]. Examples of the “non-peptidic physiologically active substance”, in particular, “a poorly water-soluble non-peptidic physiologically active substance” include substances having a receptor action or antagonistic action, an enzyme inhibitory action, a carrier promoting or suppressing action, and the like. . The receptor in which the “non-peptidic physiologically active substance”, particularly the “poorly water-soluble non-peptidic physiologically active substance” exhibits an agonizing or antagonistic action may be on the cell surface or inside the cell. The cell surface receptor includes an ion channel-linked type, a G protein-linked type and an enzyme-linked type. Types of ligands for the receptor include small peptides, proteins,
Amino acids, nucleotides, steroids, fatty acid derivatives,
There are nitric oxide, carbon monoxide and the like. Examples of the receptor include, for example, luteinizing hormone releasing hormone (LH-RH) receptor, thyroid stimulating hormone secreting hormone (TRH) receptor, corticotropin secreting hormone (CRF) receptor,
Endorphin receptor, substance P receptor, neurotensin receptor, thyroid stimulating hormone (TSH) receptor, lactating hormone (PRL) receptor, follicle stimulating hormone (FSH) receptor, luteinizing hormone (LH) receptor, adrenal cortex Stimulating hormone (ACTH) receptor and the like.

[0007] Examples of the enzymes which the "non-peptidic physiologically active substance", particularly the "poorly water-soluble non-peptidic physiologically active substance" exerts an inhibitory action include blood coagulation enzymes, fibrinolytic enzymes, digestive enzymes, and phosphorylating enzymes. , Metabolic enzymes, antioxidant enzymes and the like.
Examples of the enzyme include monoamine oxidase (MA)
O), angiotensin converting enzyme, HMG-CoA reductase, cholesterol esterification enzyme (ACAT), cyclooxygenase (COX), trypsin, α-chymotrypsin, kallikrein, β-galactosidase, elastase, thrombomodulin, thrombin, blood coagulation factors (I Factor-F), protein C, protein S, plasmin, plasminogen activator, urokinase, protein kinase C, tyrosine kinase, cytochrome p450s (3A4, 1A, 2
C, 2D, etc.), superoxide dismutase (S
OD). Examples of the enzyme which the CAD exhibits an inhibitory action include those derived from human cells, bacteria, phages or viruses. CAD showing the inhibitory action is
It is expected to have antibacterial or antiviral action. Examples of the enzyme include a cross-linking enzyme transpeptidase, a penicillin-binding protein (PBP-1A, PBP-1
B, PBP-2, PBP-3, PBP-4, PBP-
5, PBP-6), neuraminidase, aminopeptidase A, aminopeptidase B, α-amylase, β-
Camelmases, reverse transcriptase inhibitors and the like.

[0008] Carriers in which the "non-peptidic physiologically active substance", in particular, the "poorly water-soluble non-peptidic physiologically active substance" exhibits promotion or inhibition, include passive or active ion channels, glucose transporters, Peptide transporters, p-glycoprotein and the like. Examples of the carrier include voltage-gated sodium channels, calcium-gated sodium channels, potassium-gated calcium channels, potassium channels, chloride ion channels, gastric mucosal proton pumps (H ⁺ , K ⁺ -ATPas
e), glucose transporters (GLUT1, G
LUT2, GLUT3, GLUT4), PEPT1, M
DR1, MDR2, MRP, cMOAT, ACT1 and the like.

The "non-peptidic physiologically active substance", in particular, the "poorly water-soluble non-peptidic physiologically active substance" is not particularly limited as long as it has the above-mentioned activity. As an analgesic and anti-inflammatory effect,
Sulpyrin, indomethacin, atropine, scopolamine, morphine, pethidine or a salt thereof, which have a tranquilizing effect, diazepam, lorazepam, etc., have an antibacterial effect, and griseofulvin, etc., have an antibiotic effect. As dibekacin, canendomycin, ribidomycin, tobramycin, amikacin, fradiomycin, sisomicin, tetracycline, oxytetracycline, lolitetracycline, doxycycline, ampicillin, moxalactam, thienamycin, sulfazecin, azuleonal or a salt thereof, has antitumor activity Fumegillol, mitomycin C, adriamycin, fluorouracil, etc. have antihyperlipidemic action,
Clofibrate, etc. have antitussive and antitussive effects, ephedrine, methylephedrine, noscapine,
Codeine, dihydrocodeine, cloperastine, protochlorol, isoproterenol, salbutamol, terbutaline or a salt thereof, as those having a muscle relaxing action, pridinol, pancuronium, etc. Oxide etc., as those having an anti-ulcer effect, metoclopramide etc., as those having an antidepressant effect, clomipramine etc., as those having an anti-allergic effect, diphenhydramine, tripelenamine, diphenylpyraline,
Methoxyphenamine etc., as those that have a cardiotonic effect,
Ethylephrine, etc., as those with arrhythmia treatment effect,
Alprenolol, bufetrol, oxprenolol, etc., as those having a vasodilatory effect, oxyfedrine, bamethane, etc., as those having a hypotensive diuretic effect,
Pentolinium, Mecamylaluminin, Clonidine, etc.
As those having an antidiabetic effect, glybuzole etc., as those having an antituberculosis effect, ethambutol etc., as those having a narcotic antagonistic effect, levallorphan, nalorphine, naloxone or a salt thereof, etc., as those having a hormonal effect, Estrogen, luteinizing hormone (L
H) class, dexamethasone, hexestrol, betamethasone, triamcinolone, triamcinolone setonide,
Examples include fluocinolone acetonide, prednisolone, hydrocortisone, and fat-soluble vitamins such as vitamin A, vitamin D, vitamin E, vitamin K, and folic acid (vitamin M).

Examples of the "non-peptidic physiologically active substance", in particular, the "poorly water-soluble non-peptidic physiologically active substance" having the properties of CAD include, for example, amilodarone,
Examples thereof include those exhibiting a receptor antagonism such as promethazine and propranolol, those exhibiting an enzyme inhibitory effect such as chloramphenicol and gentamicin, and those exhibiting a carrier antagonism such as amitriptyline, imipramine and trimiprosine.

[0011] The "non-peptidic physiologically active substance", in particular, the "poorly water-soluble non-peptidic physiologically active substance" has a molecular weight of about 1,000 or less, preferably about 900 or less, more preferably about 800 or less. Particularly preferably about 700
The following substances are preferred: The solubility of the "poorly water-soluble non-peptide physiologically active substance" is, for example, 0.1% (W / V) or less, preferably 0.01% (W / V) or less. Here, the solubility refers to, for example, a Recipro Shaker (model SR
-I, Taiyo Kagaku Kogyo Co., Ltd.), shake at least 100 times per minute at room temperature (about 15 to about 25 ° C.) for 30 minutes.
This shows the drug concentration in the supernatant obtained by centrifuging the drug remaining undissolved after more than one minute. Said "non-peptidic bioactive substance", particularly "poorly water-soluble non-peptidic bioactive substance"
Preferably, a gonadotropin releasing hormone (GnRH (Gonadotropin releasing hormone)) agonist (agonist) or antagonist (antagonist), more preferably a GnRH antagonist.
Such GnRH is luteinizing hormone releasing hormone: L
H-RH (Luteinizing hormone-releasing hormone)
It has a similar effect. The “GnRH antagonist” may be any compound having a GnRH antagonistic action.

Embedded image Wherein, X is a carbon atom or a nitrogen atom, - - - indicates a single bond or a double bond], or a salt thereof having a partial structure represented by (basic skeleton), and more specifically, the Compound (I), compound (VIII) and the like.

The definition of each substituent in the above formula (I) is described below. “C _1-4 alkoxy group” represented by R ¹ or R ²
As, for example, methoxy, ethoxy, propoxy,
Isopropoxy, butoxy, tert-butoxy and the like. Of these, a C _1-3 alkoxy group is preferred. More preferably, it is methoxy. The “C _1-4 alkoxy-carbonyl group” represented by R ¹ or R ² includes, for example, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl and the like. Of these, a C _1-3 alkoxy-carbonyl group is preferred. More preferably, it is methoxycarbonyl. R
¹ or an optionally substituted C ^{2 represented} by R ²
_As the “C _1-4 alkyl group” of the “ _1-4 alkyl group”, for example, a linear C _1-4 alkyl group (eg, methyl, ethyl, propyl, butyl, etc.), a branched C _3-4 alkyl group (Eg, isopropyl, isobutyl, sec-butyl, tert-butyl, etc.). Of these, a C _1-3 alkyl group is preferred. Especially, ethyl is preferable.

The “substituent” of the “optionally substituted C _1-4 alkyl group” represented by R ¹ or R ² includes:
For example, (i) hydroxy, (ii) C _1-7 acyloxy (eg, C _1-6 alkyl-carbonyloxy such as acetoxy, propionyloxy), (iii) benzoyloxy, (iv) C _1-6 alkoxy-carbonyl ( examples include methoxycarbonyl, ethoxycarbonyl, tert- butoxycarbonyl), benzyloxycarbonyl, C _1-4 acyl (e.g., acetyl, C _1-3 alkyl, such as propionyl - carbonyl), C _1-4 alkyl (e.g., Methyl,
An amino group (eg, amino, dimethylamino, methoxycarbonyl) which may have one or two substituents selected from ethyl, propyl, butyl, etc.) and C _1-3 alkylsulfonyl (eg, methanesulfonyl) Amino, ethoxycarbonylamino, tert-butoxycarbonylamino, benzyloxycarbonylamino, acetylamino, methanesulfonylamino, etc.), (v) C
_1-10 alkoxy (eg, methoxy, ethoxy, propoxy, tert-butoxy, etc.), (vi) C _3-7 cycloalkyloxycarbonyloxy-C _1-3 alkoxy (eg, cyclohexyloxycarbonyloxy-1-ethoxy, etc.) , (Vii) C _1-3 alkoxy-C _1-3 alkoxy (eg, methoxymethoxy, methoxyethoxy, etc.). Of these, hydroxy is preferred. R ¹
Or a “C _1-4 optionally having substituent (s)” for R ²
The “C _1-4 alkyl group” of the “alkyl group” includes, for example, 1 to 5, preferably 1 to 5,
It may have from 3 to 3, and when the number of substituents is 2 or more, each substituent may be the same or different. One of R ¹ and R ² is preferably a hydrogen atom, and the other is preferably a C _1-3 alkoxy group.

The "halogen atom" represented by R ³ includes, for example, fluorine, chlorine, bromine and iodine. Of these, chlorine is preferred. As the "C _1-4 alkoxy group" of the "optionally C _1-4 alkoxy group which may have a substituent" represented by R ^3, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert- butoxy And the like. Of these, methoxy is preferred. R
^3, as the "substituent" of the "optionally substituted C _1-4 alkoxy group" represented, the R ¹ or R ² "which may have a substituent C ₁ represented by And the same as the "substituent" of the " _-4 alkyl group". C _1-4
Alkoxy groups are preferred. The C _1-4 alkoxy group may have, for example, 1 to 5, preferably 1 to 3 of the above substituents at substitutable positions, and the number of substituents is 2
In the case of two or more, each substituent may be the same or different. The “C _1-4 alkylenedioxy group” formed by linking two adjacent R ³ includes, for example, methylenedioxy, ethylenedioxy and the like. R ³ is preferably a hydrogen atom.

The “C _1-4 alkyl group” represented by R ⁴ includes, for example, a linear C _1-4 alkyl group (eg, methyl, ethyl, propyl, butyl, etc.), a branched C _3-4 alkyl group. Groups (eg, isopropyl, isobutyl, sec-butyl, tert-
Butyl, etc.). Among them, a C _1-3 alkyl group is preferred. Especially, methyl is preferable. Represented by R ⁶ As the "C _1-4 alkyl group optionally having a substituent" represented by R ¹ or R ² "optionally substituted C _1-4 alkyl group" Is mentioned.

The "heterocycle" formed by linking R ⁴ and R ⁵ includes a 5- or 6-membered nitrogen-containing heterocyclic group. When R ⁴ and R ⁵ are linked, the formula

Embedded image As the group represented by, for example,

Embedded image And the like. Of these, the formula

Embedded image The group represented by is preferred.

R ⁶ is represented by the formula

Embedded image [Wherein, R ⁵ has the same meaning as described above]. R ⁴ is preferably a C _1-3 alkyl group and R ⁵ is preferably a hydrogen atom. n is preferably an integer of 0 to 2.

[0018] In compound (I), preferred compounds, R ¹ is hydroxy group, a methoxy group or a C _1-3 alkyl group; R ² is a hydrogen atom or a C _1-3 alkyl group; R ⁴ is C
_A compound wherein R ⁶ is a benzyl group; and n is 0, or a salt thereof. Among them, preferred are compounds in which R ¹ is a methoxy group; R ² and R ⁵ are each a hydrogen atom; R ⁴ is a C _1-3 alkyl group; R ⁶ is a benzyl group; . Specific examples of compound (I) include 5- (N-benzyl-N
-Methylaminomethyl) -1- (2,6-difluorobenzyl) -6- [4- (3-methoxyureido) phenyl] -3-phenylthieno [2,3-d] pyrimidine-
2,4 (1H, 3H) -dione, 5- (N-benzyl-N
-Methylaminomethyl) -1- (2,6-difluorobenzyl) -6- [4- (3-hydroxyureido) phenyl] -3-phenylthieno [2,3-d] pyrimidine-2,4 (1H, 3H) -dione, 5- (N-benzyl-)
N-methylaminomethyl) -1- (2,6-difluorobenzyl) -6- [4- (3-methylureido) phenyl] -3-phenylthieno [2,3-d] pyrimidine-
2,4 (1H, 3H) -dione, 5- (N-benzyl-N
-Methylaminomethyl) -1- (2,6-difluorobenzyl) -6- [4- (3-ethylureido) phenyl] -3-phenylthieno [2,3-d] pyrimidine-
2,4 (1H, 3H) -dione or a salt thereof. Among them, 5- (N-benzyl-N-methylaminomethyl) -1- (2,6-difluorobenzyl) -6
-[4- (3-methoxyureido) phenyl] -3-phenylthieno [2,3-d] pyrimidine-2,4 (1H,
3H) -dione or a salt thereof is preferred.

The definition of each substituent in the above formula (VIII) is described below. Examples of the “C _1-7 alkyl group” of the “ _optionally substituted C _1-7 alkyl group” for R ⁹ include, for example, a linear C _1-7 alkyl group (eg, methyl, ethyl, propyl,
Butyl, pentyl, hexyl, heptyl, etc.), branched C
_3-7 alkyl group (eg, isopropyl, isobutyl, sec-
Butyl, tert-butyl, isopentyl, neopentyl and the like). Of these, a branched C _3-7 alkyl group is preferred. Especially, isopropyl is preferable. R
Indicated by ⁹ "optionally substituted C _1-7 alkyl group"
Examples of the “substituent” include (i) a hydroxy group, (i)
i) C _1-7 acyloxy (eg, C _1-6 alkyl-carbonyloxy such as acetoxy, propionyloxy; benzoyloxy, etc.); (iii) C _1-6 alkoxy-carbonyl (eg, methoxycarbonyl, ethoxycarbonyl,
tert-butoxycarbonyl, etc.), benzyloxycarbonyl, C _1-3 acyl (eg, C _1-2 alkyl-carbonyl such as acetyl, propionyl, etc.), C _1-3 alkylsulfonyl (eg, methanesulfonyl, etc.) and C _{1 -3}
Amino (eg, amino, methoxycarbonylamino, ethoxycarbonylamino, tert-butoxycarbonylamino, benzyloxycarbonylamino) optionally having one or two substituents selected from alkyl (eg, methyl, ethyl, etc.) , Acetylamino, methanesulfonylamino, methylamino, dimethylamino, etc.),
(Iv) C _3-7 cycloalkyloxycarbonyloxy (eg, cyclohexyloxycarbonyloxy, etc.) and C _1-3 alkoxy (eg, methoxy, ethoxy, etc.)
C which may have 1 to 3 substituents selected from
_1-10 (preferably C _1-4 ) alkoxy (eg, methoxy,
Ethoxy, propoxy, tert-butoxy, cyclohexyloxycarbonyloxy-1-ethoxy, methoxymethoxy, ethoxymethoxy and the like, (v) C _1-6 alkoxy-carbonyl (eg, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl and the like) and the like. No. Of these, a hydroxy group is preferred. The “C _1-7 alkyl group” is, for example, one of the above substituents at a substitutable position.
It may have from 5 to 5, preferably 1 to 3, and when the number of substituents is 2 or more, each substituent may be the same or different.

The “optionally substituted C” represented by R ⁹
_As the “C _3-7 cycloalkyl group” of the “ _3-7 cycloalkyl group”, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like can be mentioned. Of these, cyclopropyl is preferred. “Optionally substituted C _{3-7 represented} by R ⁹
Examples of the “substituent” of the “cycloalkyl group” include 1 to 3 same groups as the “substituent” of the “optionally substituted C _1-7 alkyl group” for R ⁹ . When the number of substituents is two or more, each substituent may be the same or different. Represented by R ⁹ in the "optionally substituted C _1-6 alkoxy group" as "C _1-6 alkoxy group" is, for example, mono- - or di -C _1-6 alkoxy group (eg, Methoxyamino, ethoxyamino,
Dimethoxyamino, diethoxyamino, ethoxymethoxyamino, etc.). Among them, Mono-C _1-3
An alkoxyamino group (eg, methoxyamino, etc.) is preferred. Represented by R ⁹ in the "optionally substituted C _1-6 alkoxy group" as the "substituent", the represented by R ⁹ in the "optionally substituted C _1-7 alkyl group", " And the same number as the “substituent”. When the number of substituents is two or more, each substituent may be the same or different. The "substituent" may be substituted with "C _1-6 alkoxy group" or "amino group" of the C _1-6 alkoxy amino group. Specific examples of the “optionally substituted C _1-6 alkoxyamino group” include methoxyamino, N-
Methyl-N-methoxyamino, N-ethyl-N-methoxyamino, ethoxyamino, dimethoxyamino, diethoxyamino, ethoxymethoxyamino and the like. Preferred examples include a C _1-3 alkoxyamino group,
And an N-C _1-3 alkyl-NC _1-3 alkoxyamino group.

R⁹The “optionally substituted
As the “substituent” for the “droxyamino group”, hydroxy
Substitute "hydroxy group" or "amino group" for amino group
As a substituent on the “hydroxy group”
Is (i) C_1-7Acyl group (eg, acetyl, propionyl)
Such as C_1-6Alkyl-carbonyl; benzoyl
Etc.), (ii) C_1-6Alkoxy-carbonyl (eg, meth
Xoxycarbonyl, ethoxycarbonyl, tert-butoxy
Carbonyl), benzyloxycarbonyl, C _1-3
Acyl (eg, C such as acetyl, propionyl, etc.)_1-2Al
Kill-carbonyl, etc.), C_1-3Alkylsulfonyl
(Eg, methanesulfonyl, etc.) and C_1-3Alkyl
(Eg, methyl, ethyl, etc.)
An amino group optionally having one or two amino groups (eg, amino,
Methoxycarbonylamino, ethoxycarbonylamino
, Tert-butoxycarbonylamino, benzyloxy
Carbonylamino, acetylamino, methanesulfonyl
Amino, methylamino, dimethylamino, etc.), (ii
i) C_3-7Cycloalkyloxycarbonyloxy (eg,
Cyclohexyloxycarbonyloxy) and C
_1-3Select from alkoxy (eg, methoxy, ethoxy, etc.)
C which may have 1 to 3 substituents
_1-10(Preferably C_1-4) Alkyl group (eg, methyl,
Tyl, propyl, tert-butyl, cyclohexyloxy
Carbonyloxy-1-ethyl, methoxymethyl, ethoxy
And the like, and the above “amino group”
Examples of the substituent include the groups described in (i) to (iii) above.
I can do it. "Hydroxy group" of hydroxyamino group or
May be the same or different even if each substituent of the `` amino group '' is the same or different
Good. "Optionally substituted hydroxyamino group"
Preferred examples include NC _1-6Alkyl-N-hydro
Xyamino group (eg, N-methyl-N-hydroxyamido)
And N-ethyl-N-hydroxyamino).
I can do it. More preferably N-C_1-3Alkyl-N-
And a hydroxyamino group.

The optionally substituted C represented by R ¹⁰
_As the “C _1-7 alkyl group” of the “ _1-7 alkyl group”, for example, a linear or branched C _1-7 alkyl group (eg, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, se
c-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl and the like). Among them, a C _1-3 alkyl group (eg, methyl, ethyl,
Propyl, isopropyl, etc.). Particularly preferred is isopropyl. As the “substituent” of the “optionally substituted C _1-7 alkyl group” for R ¹⁰ ,
The same number as the “substituent” of the “optionally substituted C _1-7 alkyl group” for R ⁹ is mentioned.
When the number of substituents is two or more, each substituent may be the same or different. Examples of the “substituent” of the “optionally substituted phenyl group” represented by R ¹⁰ include halogen (eg, fluorine, chlorine, bromine, iodine, etc.), C _1-3 alkyl group (eg, methyl, ethyl , Propyl, isopropyl, etc.) and C _1-3 alkoxy groups (eg, methoxy, ethoxy, propoxy, isopropoxy, etc.). Among them, halogen (preferably fluorine) is preferable.
The "phenyl group" may have, for example, 1 to 5, preferably 1 to 3 of the above substituents at substitutable positions. When the number of the substituents is 2 or more, each substituent is the same Or they may be different.

R ⁹ is preferably a substituted branched C _3-7
An alkyl group or a substituted C _3-7 cycloalkyl group,
More preferably, a branched C substituted with a hydroxy group
A C _3-7 cycloalkyl group substituted with a _3-7 alkyl group or a hydroxy group. Of these, a C _3-7 cycloalkyl group substituted with a hydroxy group is preferred. Furthermore, hydroxy group optionally substituted C _1-3 also be an alkyl group, optionally substituted by a hydroxy group C _3-7 cycloalkyl group, mono--C _1-3 alkoxy amino, N-C _1-3 Alkyl-N-hydroxyamino groups, hydroxyamino groups and the like are also preferred. R ⁹ is particularly preferably a cyclopropyl group or a methoxyamino group which may be substituted with a hydroxy group. Most preferred is a cyclopropyl group substituted with a hydroxy group. R ¹⁰ is preferably an optionally substituted C _1-7 alkyl group. More preferred are a C _1-3 alkyl group which may be substituted with a hydroxy group. Particularly preferred is isopropyl. Also, phenyl is preferable. Compound (VII
As preferred examples of I), R ⁹ is a C _1-3 alkyl group optionally substituted with a hydroxy group, a C _3-7 cycloalkyl group optionally substituted with a hydroxy group, or mono-C _{1-. 3} alkoxyamino group; a compound in which R ¹⁰ is a C _1-3 alkyl group or a phenyl group, or a salt thereof. More preferably, R ⁹ is (1) a C _1-3 alkyl group substituted with one or two hydroxy groups, (2) a C _3-7 cycloalkyl group substituted with a hydroxy group, or (3) a C _3-7 cycloalkyl group. _1-3 alkoxyamino group; a compound wherein R ¹⁰ is an isopropyl group or phenyl, or a salt thereof.

Specific examples of the compound (VIII) include 3-
(N-benzyl-N-methylaminomethyl) -4,7-
Dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- (4-cyclopropanecarbonylaminophenyl) -4-oxothieno [2,3-b] pyridine, 5-benzoyl-3- (N- Benzyl-N-methylaminomethyl) -7- (2,6-difluorobenzyl) -4,7-dihydro-4-oxo-2- [4- (3
-Hydroxy-2-methylpropionylamino) phenyl] thieno [2,3-b] pyridine, 5- (4-fluorobenzoyl) -3- (N-benzyl-N-methylaminomethyl) -7- (2,6 -Difluorobenzyl)-
4,7-dihydro-4-oxo-2- (4-cyclopropanecarbonylaminophenyl) thieno [2,3-b]
Pyridine, 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7-
(2,6-difluorobenzyl) -2- [4- (3-hydroxy-2-methylpropionylamino) phenyl]
-4-oxothieno [2,3-b] pyridine, 3- (N
-Benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- (4-N'-methoxyureidophenyl) -4-oxothieno [2 3-b] pyridine, 3-
(N-benzyl-N-methylaminomethyl) -4,7-
Dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4-[(1-hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothieno [2,3-b] pyridine, (R ) -4,7-Dihydro-2- [4- (3-hydroxy-2-methylpropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl-N-methylaminomethyl ) -5-Isobutyryl-4-oxothieno [2,3-
b] pyridine, 4,7-dihydro-2- [4- (2-hydroxy-2-methylpropionylamino) phenyl]
-7- (2,6-difluorobenzyl) -3- (N-benzyl-N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2,3-b] pyridine, 4,7-
Dihydro-2- [4- (3-hydroxy-3-methylbutyrylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N-benzyl-N-methylaminomethyl) -5-isobutyryl -4-oxothieno [2, 3
-B] pyridine, (R) -4,7-dihydro-2- [4
-(2,3-dihydroxypropionylamino) phenyl] -7- (2,6-difluorobenzyl) -3- (N
-Benzyl-N-methylaminomethyl) -5-isobutyryl-4-oxothieno [2,3-b] pyridine, 3-
(N-benzyl-N-methylaminomethyl) -5-benzoyl-7- (2,6-difluorobenzyl) -4,7
-Dihydro-2- [4-[(1-hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothieno [2,3-b] pyridine or a salt thereof. Above all, 3- (N-benzyl-N-methylaminomethyl) -4,7-dihydro-5-isobutyryl-
7- (2,6-difluorobenzyl) -2- [4-
[(1-Hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothieno [2,3-b] pyridine or a salt thereof is preferred.

The salts of compound (I) and compound (VIII) are preferably physiologically acceptable acid addition salts. Examples of such a salt include salts with inorganic acids (eg, hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc.) and organic acids (eg, formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, etc.). Acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, p-
For example, a salt with toluenesulfonic acid or the like is used. When the compound (I) has an acidic group, an inorganic base (eg, an alkali metal salt or an alkaline earth metal such as sodium, potassium, calcium, magnesium or the like, ammonia or the like) or an organic base (eg, trimethylamine,
Physiologically acceptable salts may be formed with triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, N, N'-dibenzylethylenediamine, and the like.

Compound (I) can be prepared by a method known per se, for example, in JP-A-9-169768, WO 96/24
No. 597, or a method analogous thereto. Specific examples include the following production method 1 and production method 2. The compounds in the reaction formulas include those forming salts, and examples of the salts include those similar to the salts of compound (I).

Embedded image In the above formula, L represents a leaving group, and other symbols have the same meanings as described above. As the "leaving group" represented by L, for example, 1
-Imidazolyl, a halogen atom, an optionally substituted alkoxy group, and the like. The “alkoxy group optionally having substituent (s)” includes a C _1-4 alkoxy group optionally having 1 to 3 halogen atoms (eg, chlorine, bromine, etc.) (eg, 2, 2,2-trichloroethoxy group).

Compound (II) is disclosed in JP-A-9-169768.
The method can be obtained by a method described in Japanese Patent Application Laid-Open Publication No. H10-205 or a method analogous thereto. Compound (II) and carbonyldiimidazole (N,
Reaction with N′-carbonyldiimidazole; CDI) or phosgene (including dimers and trimers) to obtain compound (IV), and then reaction with compound (III) to obtain compound (I) . The compound (IV) may be allowed to continue the reaction without isolation, or may be isolated and used in the next step.
Compound (IV) can also be obtained by reacting compound (II) with a chloroformate compound (eg, 2,2,2-trichloroethyl chloroformate, 1-chloroethyl chloroformate, etc.). .

In the reaction of compound (II) with carbonyldiimidazole or phosgene, the amount of carbonyldiimidazole or phosgene used is the same as that of compound (II) 1
It is about 1 to 3 moles per mole. The reaction is
The reaction is usually performed in a suitable solvent that does not adversely influence the reaction. Examples of the solvent include ethers (eg, ethyl ether, dioxane, dimethoxyethane, tetrahydrofuran, etc.), aromatic hydrocarbons (eg, benzene, toluene, etc.), amides (eg, dimethylformamide,
Dimethylacetamide), halogenated hydrocarbons (eg, chloroform, dichloromethane, etc.) and the like are used. The reaction temperature is usually about 0 to about 150 ° C, preferably at room temperature (about 15 to about 25 ° C). The reaction time is usually about 1 to about 36 hours. This reaction is performed, if necessary, in the presence of a base. As the "base", for example,
Sodium carbonate, sodium bicarbonate, potassium carbonate,
An inorganic base such as potassium hydrogen carbonate, sodium hydroxide, potassium hydroxide, and thallium hydroxide, or an organic base such as triethylamine and pyridine is used. The amount of the “base” to be used is about 2 to 1 mol of the compound (II).
Mol to 20 mol, preferably about 5 mol to 12 mol. The subsequent reaction conditions with compound (III) may be the same as those for reacting compound (II) with carbonyldiimidazole or phosgene. Compound (II
The amount of I) to be used is about 2 to 20 mol, preferably about 5 to 10 mol, per 1 mol of compound (II) or compound (IV). The reaction temperature is usually about 0 to 150 ° C,
Preferably at room temperature (about 15 to 25 ° C). The reaction time is usually about 1 to 6 hours. Further, carbonyldiimidazole or phosgene and compound (III) may be simultaneously reacted with compound (II).

[0029]

Embedded image In the above formula, R ⁷ is a hydrogen atom or an alkyl group, R ⁸ is an alkyl group, and other symbols are as defined above. R
⁷ or as "alkyl group" represented by R ^8, similar to the "C _1-4 alkyl group" of the "optionally substituted C _1-4 alkyl group" represented by R ¹ or R ² One.

The compound (V) is reacted with a method known per se, for example, by reacting p-nitrophenylacetone, a cyanoacetate derivative and sulfur (eg, Chem. Ber., Vol. 99, 94-1).
00, 1966), resulting 2-amino-4-methyl-
5- (4-nitrophenyl) thiophene is disclosed in
169768, WO 96/24597 and the like or a method analogous thereto.

When R ⁷ is a hydrogen atom, compound (V) is
In the presence of a condensing reagent, the formula

Embedded image Wherein each symbol is as defined above, or a salt thereof (hereinafter abbreviated as compound (VI)) to obtain compound (VII), which is then subjected to a ring closure reaction. ,
Compound (I) is obtained. Examples of the "condensing reagent" include benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate (benz
otriazol-1-yloxytripyrrolidinophosphonium hexafluo
rophosphate: PyBOP). The amount of the “condensing reagent” to be used is about 1 to 3 with respect to 1 mol of compound (V).
Is a mole. This reaction is generally performed in a suitable solvent that does not adversely influence the reaction. Examples of the solvent include alcohols (eg, ethanol, methanol, etc.), aromatic hydrocarbons (eg, benzene, toluene, etc.), amides (eg, dimethylformamide, dimethylacetamide, etc.), halogenated hydrocarbons (Eg, chloroform, dichloromethane, etc.) are used. The reaction temperature is generally about 0 to about 150 ° C, preferably at room temperature (about 15 to about 150 ° C).
About 25 ° C). The reaction time is usually about 1 to about 36 hours. The product can be used in the next reaction as a reaction solution or as a crude product, but can also be isolated from the reaction mixture according to a conventional method.

Compound (VII) is subjected to a ring closure reaction in the presence of a base. Examples of the "base" include inorganic bases such as sodium methoxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, sodium hydroxide, potassium hydroxide, and thallium hydroxide; and organic bases such as triethylamine and pyridine. A base is used. The amount of the “base” to be used is about 2 to 20 mol, preferably about 5 to 12 mol, per 1 mol of compound (VII).
Is a mole. This reaction is generally performed in a suitable solvent that does not adversely influence the reaction. Examples of the solvent include alcohols (eg, ethanol, methanol, etc.), aromatic hydrocarbons (eg, benzene, toluene, etc.), amides (eg, dimethylformamide, dimethylacetamide, etc.), halogenated hydrocarbons (Eg, chloroform, dichloromethane, etc.) are used. The reaction temperature is generally about 0 to about 150 ° C, preferably at room temperature (about 15 to about 150 ° C).
About 25 ° C). The reaction time is usually about 1 to about 36 hours.

When R ⁷ is an alkyl group, compound (V)
Is reacted with the activated compound (VI) to obtain a compound (I). The activated compound (VI) can be produced according to a method known per se. For example, an organic aluminum reagent and a compound (VI) can be produced in a suitable solvent that does not adversely influence the reaction.
And by reacting As the “organoaluminum reagent”, for example, trimethylaluminum, dimethylaluminum chloride and the like, or a solution containing these and the like can be mentioned. The amount of the “organoaluminum reagent” used is 1 mole per 1 mole of compound (VI).
５5 mol, preferably 1 mol. As the solvent,
For example, halogenated hydrocarbons (eg, chloroform, dichloromethane, etc.) are preferred. The reaction temperature is usually about 0
To 150 ° C., preferably at room temperature (about 15 to 25 ° C.). The reaction time is usually about 1 to 6 hours. By reacting the compound (V) with the activated compound (VI), a ring-closing reaction is performed to obtain a compound (I).
The amount of the "compound (V)" to be used is preferably about 1/5 of the mixture of the compound (VI) and the organoaluminum reagent. This reaction is generally performed in a suitable solvent that does not adversely influence the reaction. As the solvent, the solvent used in the reaction for obtaining the activated compound (VI) is preferable. The reaction temperature is generally about 0-150 ° C, preferably at room temperature (about 15-25 ° C). The reaction time is usually about 1 to 48 hours.

Compound (I) can be isolated and purified by a known separation means, for example, recrystallization, distillation, chromatography and the like. When the compound (I) is obtained in a free form, it can be converted to a target salt by a method known per se or a method analogous thereto. Alternatively, it can be converted into a free form or another desired salt by a method analogous thereto. Compound (I) may be a hydrate or a non-hydrate. Examples of the hydrate include monohydrate, 1.5-hydrate and dihydrate. When the compound (I) is obtained as a mixture of optically active compounds, it can be separated into the desired (R) -form or (S) -form by a known optical resolution means. Compound (I) is an isotope (eg, ³ H, ¹⁴ C, ³⁵ S)
It may be labeled with, for example.

Compound (VIII) or a salt thereof can be prepared by a method known per se, for example, WO 95/28405, WO
It can be produced by the method described in JP 00/00493 or a method analogous thereto.

The "biodegradable polymer having two or more carboxyl groups at the end" used in the present invention includes a biodegradable polymer having two or more, preferably two or three carboxyl groups at the end. Is mentioned.
Among them, a biodegradable polymer having an α, α-dicarboxyl group or α, β, β′-tricarboxyl group at the terminal is preferable. The “terminal” refers to the minimum repeating unit (minimum repeating unit) which is bonded to the adjacent minimum repeating unit only on one side among the minimum repeating units constituting the polymer. Further, when the main chain of the polymer has a hetero atom, the terminal generated on the left side is referred to as α residue, and the terminal generated on the right side is referred to as ω residue, when the minimum repeating unit is preferentially described from left to right. A linear polymer has two terminals, and a branched polymer such as a comb polymer or a star polymer has two or more terminals.

Examples of the “biodegradable polymer having two or more carboxyl groups at the terminal” include, for example, those in which the ω residue is a hydroxypolycarboxylic acid (eg, tartronic acid,
-Hydroxyethylmalonic acid, malic acid, citric acid, etc.), and the portion other than the ω residue is an aliphatic polyester [eg, α-hydroxycarboxylic acids (eg, glycolic acid, lactic acid, 2-hydroxybutyric acid, etc.); one or two of α-hydroxydicarboxylic acids (eg, malic acid, etc.), α-hydroxytricarboxylic acids (eg, citric acid, etc.)
Polymers or copolymers synthesized from more than one species], poly (α-cyanoacrylate), polyamino acids [eg, poly (γ-benzyl-L-glutamic acid), etc.], maleic anhydride copolymer (Eg, a styrene-maleic acid copolymer). The bonding mode of the monomer may be any of random, block, and graft. When the α-hydroxymonocarboxylic acids, α-hydroxydicarboxylic acids, and α-hydroxytricarboxylic acids have an optically active center in the molecule, any of D-, L-, and DL-forms may be used. The ω residue is preferably tartronic acid, citric acid or 2-hydroxyethylmalonic acid, more preferably tartronic acid or citric acid. The portion other than the ω residue is preferably poly α-hydroxycarboxylic acid. Among them, linear α-hydroxycarboxylic acid is preferred.

As the α-hydroxycarboxylic acid which is the minimum repeating unit of the “polyα-hydroxycarboxylic acid”, for example, lactic acid, glycolic acid and the like are preferable. As the “poly α-hydroxycarboxylic acid”, for example,
Copolymers of lactic acid, glycolic acid and the like [hereinafter may be referred to as poly (lactide-co-glycolide), poly (lactic acid-co-glycolic acid) or lactic acid-glycolic acid polymer. ]. The "lactic acid-glycolic acid polymer" means homopolymers (polymers, polylactides or polyglycolides) and copolymers (copolymers) of lactic acid and glycolic acid. The composition ratio of lactic acid and glycolic acid (lactic acid / glycolic acid; mol / mol%) in the “lactic acid-glycolic acid polymer” is not particularly limited as long as the object of the present invention is achieved. ~ 30/70, preferably about 100/0
About 40/60, more preferably about 100/0 to about 45
/ 55. Α-hydroxycarboxylic acid that is the minimum repeating unit of the “poly α-hydroxycarboxylic acid” is
When the molecule has an optically active center, it may be any of D-form, L-form and D, L-form. For example, D-body /
L-form (mol / mol%) is about 75/25 to about 25/7
5, preferably from about 60/40 to about 30/70.
Hydroxycarboxylic acids are used. Preferred examples of the “biodegradable polymer having two or more carboxyl groups at the terminal” include lactic acid-glycolic acid having an α, α-dicarboxyl group or an α, β, β′-tricarboxyl group at the terminal Polymers. More preferably, a lactic acid-glycolic acid polymer in which the ω residue is tartronic acid, a lactic acid-glycolic acid polymer in which the ω residue is citric acid, and the like are included. A good example of the above "lactic acid-glycolic acid" is polylactic acid.

The weight-average molecular weight of the “biodegradable polymer having two or more carboxyl groups at the terminal” is usually about 200 to about 100,000, preferably about 300.
To about 50,000, more preferably from about 500 to about 1
000. The degree of dispersion (weight average molecular weight / number average molecular weight) of the “biodegradable polymer having two or more carboxyl groups at the terminal” is usually about 1.1 to about 4.
0, preferably about 1.2-3.5. The "2 at the end
When the ω residue of the “biodegradable polymer having two or more carboxyl groups” is an α, α-dicarboxyl group,
The amount of terminal carboxyl groups per unit mass of the polymer is
Usually, it is about 30 to about 20,000 μmol / g, preferably about 60 to about 5,000 μmol / g, more preferably about 100 to about 1,000 μmol / g.

The above "weight average molecular weight", "number average molecular weight" and "dispersion degree" mean that the weight average molecular weight is 455,
645, 354,000, 98,900, 66,43
7, 37, 200, 17, 100, 9, 830, 5, 8
The molecular weight in terms of polystyrene measured by gel permeation chromatography (GPC) and the calculated degree of dispersion were measured using 11 kinds of monodisperse polystyrenes of 70, 2,500, 1,303 and 504 as reference substances. The measurement was performed using a high-speed GPC device (manufactured by Tosoh, HLC-8120GP).
C), GPC column KF804L × 2 (manufactured by Showa Denko) is used, and chloroform is used as a mobile phase. The term "amount of terminal carboxyl group" means a value determined by a terminal group quantification method by a labeling method. Specifically, in the case of a polymer whose ω residue is tartronic acid, a biodegradable polymer (Wmg) is dissolved in a 5N HCl / acetonitrile (v / v = 4/96) mixed solution (2 ml),
1M o-nitrophenylhydrazine (ONPH) solution (5N HCl / acetonitrile / ethanol = 1.0
2/35/15) (2 ml) and 0.15 M EDC
Solution (pyridine / ethanol = 4v / 96v) (2m
After l) was added and reacted at 40 ° C. for 30 minutes, the solvent was distilled off. After washing the residue with water (4 times), acetonitrile (2
ml), a 0.5 mol / l ethanolic potassium hydroxide solution (1 ml) is added, and the mixture is reacted at 60 ° C. for 30 minutes. Dilute the reaction with 1.5N NaOH and add Yml
And 544 nm absorbance A (/ cm) is measured for 1.5 N NaOH. On the other hand, the amount of free carboxyl groups (C mo
1 / L) was determined by NaOH titration, and 544 nm when tartronic hydrazide was obtained by the ONPH labeling method.
Assuming that the absorbance is B (/ cm), the amount of free carboxyl groups [COOH] of the polymer whose ω residue is tartronic acid
Is obtained by the following equation. [COOH] (mol / g) = (AYC) / (WB) Further, the biodegradable polymer is dissolved in a mixed solvent of toluene-acetone-methanol, and this solution is dissolved in an alcoholic potassium hydroxide solution using phenolphthalein as an indicator. And the amount of terminal carboxyl groups can be calculated.

"Salt" of "biodegradable polymer having two or more carboxyl groups at the terminal" used in the present invention
Examples thereof include salts with inorganic bases (eg, alkali metals such as sodium and potassium, alkaline earth metals such as calcium and magnesium), organic bases (eg, organic amines such as triethylamine, and basic acids such as arginine). Salts with amino acids, etc., transition metals (eg, zinc, iron, copper, etc.)
And complex salts thereof.

"Having two or more carboxyl groups at the terminal
Biodegradable polymer or a salt thereof '' is known per se.
It is manufactured by a method or a method analogous thereto. example
For example, in vivo fractions with two or more carboxyl groups at the ω end
In the case of a degradable polymer, two or more
A biodegradable polymer having a voxyl group is
Deprotection reaction after obtaining according to the method of (1) or (2)
Attached to (1) Hydroxypolycar with protected carboxyl group
Cyclic ester using a polymerization catalyst in the presence of a boronic acid derivative
The compound is subjected to a polymerization reaction, and two or more
A polymer having a carboxyl group is obtained. The "carboki
Hydroxypolycarboxylic acid derivative with protected silyl group "
Means, for example, that a carboxyl group (—COOH) is an amide
(-CONH_Two) Or ester (—COOR)¹¹)
Hydroxypolycarboxylic acid derivatives and the like
Can be In particular, carboxyl groups are esterified
Hydroxypolycarboxylic acid derivatives are preferred.
R¹¹Is, for example, C_1-6Alkyl groups (eg, methyl, ethyl
Propyl, isopropyl, butyl, tert-butyl
Etc.), C_3-8Cycloalkyl group (eg, cyclopentyl,
Cyclohexyl), C_6-12Aryl group (eg, phenyl
, Α-naphthyl, etc.), C_7-14Aralkyl groups (eg,
Phenyl-C such as benzyl and phenethyl _1-2Alkyl
A group; α-naphthyl-C such as α-naphthylmethyl_1-2A
Alkyl group). Among them, tert-butyl group,
And an benzyl group. The "carboxyl group is protected
Preferred Specific Examples of "Derived Hydroxypolycarboxylic Acid Derivatives"
Examples are dibenzyl tartronate, 2-hydroxy
Di-tert-butyl ethylmalonate, tribenzyl citrate
And the like.

Examples of the "polymerization catalyst" include, for example, organotin catalysts (eg, tin octylate, di-n-dilaurate).
Butyltin, tetraphenyltin, etc.), aluminum-based catalysts (eg, triethylaluminum, etc.), and zinc-based catalysts (eg, diethylzinc, etc.). Of these, an aluminum-based catalyst and a zinc-based catalyst are preferred, and a zinc-based catalyst is more preferred. As the solvent for the polymerization catalyst, benzene, hexane, toluene and the like are used, and among them, hexane, toluene and the like are preferable.

The "cyclic ester compound" includes, for example,
A cyclic compound having at least one ester bond in the ring. Specific examples include cyclic monoester compounds (lactones) and cyclic diester compounds (lactides). Examples of the “cyclic monoester compound” include 4-membered lactones (eg, β-propiolactone, β-butyrolactone, β-isovalerolactone,
-Caprolactone, β-isocaprolactone, β-methyl-β-valerolactone, etc.), 5-membered lactone (eg,
γ-butyrolactone, γ-valerolactone, etc.), 6-membered lactone (eg, δ-valerolactone), 7-membered lactone (eg, ε-caprolactone), p-dioxanone, 1,5-dioxepane-2- On and the like. As the “cyclic diester compound”, for example,

Embedded image Wherein, R ¹² and R ¹³ are the same or different and each is a hydrogen atom or a C _1-6 alkyl group shown (e.g., methyl, ethyl, propyl, isopropyl, butyl, tert- butyl, etc.)] represented by And the like. Among them, a compound in which R ¹² is a hydrogen atom and R ¹³ is a methyl group, or a compound in which R ¹² and R ¹³ are a hydrogen atom is preferable. Specifically, for example, glycolide, L-lactide,
D-lactide, DL-lactide, meso-lactide,
3-methyl-1,4-dioxane-2,5-dione (including an optically active substance) and the like.

Examples of the "polymerization reaction" method include a bulk polymerization method in which a reactant is melted or a solution polymerization method in which a reactant is dissolved in an appropriate solvent. The "solvent"
For example, benzene, toluene, xylene, decalin, dimethylformamide and the like are used, and among them, toluene and xylene are preferable. Although the polymerization temperature is not particularly limited, in the case of bulk polymerization, the temperature is not lower than the temperature at which the reaction product is brought into a molten state at the start of the reaction, and is usually 10 or more.
0 to 300 ° C., and in the case of solution polymerization, usually room temperature to 15 ° C.
When the temperature is 0 ° C. and the reaction temperature exceeds the boiling point of the reaction solution, the reaction may be carried out by refluxing with a condenser or in a pressure vessel. The polymerization time is appropriately determined in consideration of the polymerization temperature, other reaction conditions, physical properties of the target polymer, and the like, and is, for example, 10 minutes to 72 hours. After the reaction, if necessary, the reaction mixture is dissolved in an appropriate solvent (eg, acetone, dichloromethane, chloroform, etc.), and the polymerization is terminated with an acid (eg, hydrochloric acid, acetic anhydride, trifluoroacetic acid, etc.). If a polymer having a carboxyl group protected at the ω end is isolated by mixing it in a solvent that does not dissolve the target substance (eg, alcohol, water, ether, isopropyl ether, etc.) by a conventional method and isolating the polymer, Good.

(2) A hydroxypolycarboxylic acid derivative having a protected carboxyl group and a biodegradable polymer obtained by a method known per se (eg, a non-catalytic dehydration polycondensation method) are optionally combined with a dehydrating agent. And / or 2) protected at the ω end by a condensation reaction using
A polymer having more than one carboxyl group is obtained. The carboxyl group involved in the “condensation reaction” may be activated by a known means, or the carboxyl group may be activated during the condensation reaction. The activation may be performed by a method known per se, for example, an activated ester [eg, a substituted phenol (eg, pentachlorophenol, 2,4,4,
5-trichlorophenol, 2,4-dinitrophenol, p-nitrophenol, etc., N-substituted imides (eg, N-hydroxy-5-norbornene-2,3-dicarboximide, N-hydroxysuccinimide, N
-Hydroxy-1,2,3-benzotriazole, etc.), a method for forming a carboxylic anhydride, azide, or the like with respect to a carboxylic acid as a raw material, an acid chloride method, an oxidation-reduction method (Mukoyama method), and mixing. Acid anhydride method, N, N '
Dicyclohexylcarbodiimide method, N, N′-dicyclohexylcarbodiimide-additive method, method using Woodward reagent K, benzotriazole-1-
Examples thereof include a method using yl-oxy-tris (dimethylamino) -phosphonium hexafluorophosphate (BOP reagent). The “condensation reaction” is usually performed in a solvent that does not inhibit the reaction. Such solvents include amides (eg, dimethylformamide, etc.), ethers (eg, tetrahydrofuran, dioxane, etc.), halogenated hydrocarbons (eg, dichloromethane, chloroform, etc.), sulfoxides (eg, dimethyl sulfoxide, etc.) ), Esters (eg, ethyl acetate, etc.), N-methylpyrrolidone, N-methylmorpholine and the like. The reaction temperature is preferably from about -30C to about 50C, more preferably from about 0 to about 40C. The reaction time is, for example, about 10 minutes to about 24 hours.

A method for subjecting the “biodegradable polymer having two or more carboxyl groups protected at the ω-terminal” obtained in the above (1) or (2) to a “deprotection reaction” is known per se. Method. Any method capable of removing the protecting group without affecting the ester bond of the poly (hydroxycarboxylic acid) may be used, and examples thereof include a reduction reaction and an acid decomposition reaction. Examples of the “reduction reaction” include catalytic reduction using a catalyst (eg, palladium carbon, palladium black, platinum oxide, etc.), reduction with sodium in liquid ammonium, reduction with dithiothreitol, and the like. For example, when catalytically reducing a polymer having a carboxyl group protected by a benzyl group at the ω end, specifically, such a polymer is dissolved in ethyl acetate, dichloromethane, chloroform, or the like, and then palladium carbon is added thereto, followed by vigorous stirring. While bubbling hydrogen at room temperature for about 20 minutes to about 4 hours.

The "acid decomposition reaction" includes, for example, inorganic acids (eg, hydrogen fluoride, hydrogen bromide, hydrogen chloride, etc.), organic acids (eg, trifluoroacetic acid, methanesulfonic acid, trifluoromethanesulfonic acid, etc.). ) Or a mixture thereof and the like. If necessary, a cation scavenger (eg, anisole, phenol, thioanisole, etc.) may be appropriately added during acid decomposition. For example, when a polymer having a carboxyl group protected by a tert-butyl group at the ω end is acid-decomposed, specifically, such a polymer is dissolved in dichloromethane, xylene, toluene or the like, and then an appropriate amount of trifluoroacetic acid is added. ,
Alternatively, the polymer is dissolved in trifluoroacetic acid and stirred at room temperature for about 1 hour. The acid decomposition reaction is preferably
It is performed immediately after the polymerization reaction of the above (1). In this case, it can also serve as a polymerization termination reaction. Further, if necessary, the biodegradable polymer having a free carboxyl group at the ω-terminal obtained by the above deprotection reaction is subjected to an acid hydrolysis reaction to obtain a weight average molecular weight and a number average molecular weight of the polymer. Alternatively, the amount of terminal carboxyl groups can be adjusted according to the purpose. Specifically, for example, EP-A-
The method may be performed by the method described in JP 0839525 or a method analogous thereto.

The content of the “non-peptidic physiologically active substance” in the “composition” of the present invention, in particular, the “sustained release composition” is, for example, about 0.1 to about 90% (w / w), preferably Is about 0.
5 to about 80% (w / w), more preferably about 1 to 70%
(W / w). The content of the "biodegradable polymer having two or more carboxyl groups at the terminal or a salt thereof" in the "composition" of the present invention, particularly the "sustained release composition" is, for example, about 10 to about 99. 9% (w / w), preferably about 2
0 to about 95.5% (w / w), more preferably about 30 to
About 99% (w / w). The quantitative ratio of the non-peptidic bioactive substance to the biodegradable polymer having two or more carboxyl groups at the terminal or a salt thereof is, for example, 1 mol of the non-peptidic bioactive substance and the polymer or its salt. About 1/20 to about 100 moles, preferably about 1/10
To about 50 moles, more preferably about 1/5 to about 10 moles.

The “biodegradable polymer having two or more carboxyl groups at the terminal” may be used as a mixture with a biodegradable polymer having a monocarboxyl terminal.
Examples of the “biodegradable polymer having a terminal monocarboxy group” include polyα-hydroxycarboxylic acid. Examples of the “polyα-hydroxycarboxylic acid” include homopolymers (polymers, polylactides or polyglycolides) and copolymers (copolymers) such as lactic acid and glycolic acid.
The weight ratio of the “biodegradable polymer having two or more carboxyl groups at the terminal” and the “biodegradable polymer having a terminal monocarboxyl group” is, for example, 100: 0 to 0.
The ratio is 30:70, preferably 100: 0 to 50:50.

The "composition" of the present invention, particularly the "sustained release composition", can be produced by a known means, for example, an in-water drying method, a phase separation method, a spray drying method or a method analogous thereto. Hereinafter, a production method when the composition is a sustained release microcapsule (which may be referred to as a microsphere), which is a preferable example, will be described. During the following production process, if necessary, a drug holding agent (eg, gelatin, hydroxynaphthoic acid, salicylic acid, etc.) may be added by a method known per se. (I) Drying in Water An organic solvent solution containing a biodegradable polymer having two or more carboxyl groups at its terminals or a salt thereof (hereinafter abbreviated as “biodegradable polymer”) is obtained. As the organic solvent, those having a boiling point of 120 ° C. or less are preferable. For example, halogenated hydrocarbons (eg, dichloromethane, chloroform, dichloroethane, trichloroethane, carbon tetrachloride, etc.), ethers (eg, ethyl ether, isopropyl Ethers), fatty acid esters
(Eg, ethyl acetate, butyl acetate, etc.), aromatic hydrocarbons (eg, benzene, toluene, etc.), alcohols (eg, ethanol, methanol, etc.), acetonitrile, and mixtures thereof. Of these, halogenated hydrocarbons are preferred, and dichloromethane is more preferred. As the mixture, a mixture of a halogenated hydrocarbon and an alcohol is preferable, and a mixture of dichloromethane and ethanol is preferable. The concentration of the biodegradable polymer in the `` organic solvent solution containing the biodegradable polymer '' differs depending on the molecular weight of the polymer and the type of the organic solvent.For example, when dichloromethane is used as the organic solvent, It is about 0.5 to about 80% by weight, preferably about 1 to about 70% by weight, more preferably about 2 to about 60% by weight. A non-peptidic physiologically active substance (hereinafter abbreviated as “physiologically active substance”) is dissolved or dispersed in the obtained organic solvent solution. The upper limit of the weight ratio of the physiologically active substance to the biodegradable polymer is about 1: 0.2, preferably about 1: 0.5.

A solubilizing agent for increasing the solubility of the physiologically active substance may be added to the organic solvent solution of the biodegradable polymer. As the “dissolution aid”, for example, a substance which is acidic and soluble in an organic solvent solution containing a polymer can be mentioned. Of these, acetic acid and hydroxycarboxylic acids having a benzene ring (eg, salicylic acid, 3-hydroxy-2-naphthoic acid, 1-hydroxy-2-naphthoic acid, pamoic acid, etc.) are preferred. Among them, salicylic acid, 3-hydroxy-2-naphthoic acid, 1
-Hydroxy-2-naphthoic acid and the like. More preferred are salicylic acid and 3-hydroxy-2-naphthoic acid.

Next, the organic solvent solution containing the obtained composition containing the biodegradable polymer and the physiologically active substance was added to the aqueous phase to form an O (oil phase) / W (aqueous phase) emulsion. Thereafter, the solvent in the oil phase is evaporated to prepare microcapsules. In this case, the volume of the aqueous phase is generally about 1 to about 10,000 times, preferably about 5 to about 5,000 times, more preferably about 10 to about 2, times the volume of the oil phase.
000 times. In addition to the above, an emulsifier may be added to the aqueous phase. The “emulsifier” may be any one that can form a generally stable O / W emulsion. Specific examples include anionic surfactants (eg, sodium oleate, sodium stearate, sodium lauryl sulfate, etc.), nonionic surfactants (eg, polyoxyethylene sorbitan fatty acid esters [Tween
n) 80, Tween 60, Atlas Powder Co., Ltd.], polyoxyethylene castor oil derivative [HCO
-60, HCO-50, Nikko Chemicals], polyvinylpyrrolidone, polyvinyl alcohol, carboxymethylcellulose, lecithin, gelatin, hyaluronic acid and the like. These emulsifiers may be used alone or in combination of two or more. The concentration of the "emulsifier" is
For example, about 0.01 to 10% by weight, preferably about 0.05
~ 5% by weight.

In addition to the above, an osmotic pressure regulator may be added to the aqueous phase. The "osmotic pressure adjusting agent" may be any one that exhibits an osmotic pressure when it is made into an aqueous solution, and includes, for example, polyhydric alcohols, monohydric alcohols, monosaccharides, disaccharides, oligosaccharides, and amino acids or the like. And the like. As the "polyhydric alcohols", for example,
Examples include trihydric alcohols such as glycerin, pentahydric alcohols such as arabitol, xylitol and adonitol, and hexahydric alcohols such as mannitol, sorbitol and dulcitol. Of these, hexahydric alcohols are preferred, and mannitol is more preferred. The "monohydric alcohols" include, for example, methanol, ethanol, isopropyl alcohol and the like, among which ethanol is preferable. Examples of the “monosaccharide” include pentoses such as arabinose, xylose, ribose, and 2-deoxyribose, and hexoses such as glucose, fructose, galactose, mannose, sorbose, rhamnose, and fucose. Hexasaccharides are preferred.
Examples of the “oligosaccharide” include maltotriose,
Examples include trisaccharides such as raffinose sugar and tetrasaccharides such as stachyose, and among them, trisaccharides are preferable. The monosaccharide,
Examples of “derivatives” of disaccharides and oligosaccharides include, for example,
Glucosamine, galactosamine, glucuronic acid, galacturonic acid and the like are used. As the "amino acids", any L-form can be used,
For example, glycine, leucine, arginine and the like can be mentioned. Of these, L-arginine is preferred. These osmotic pressure adjusting agents may be used alone or in combination of two or more. The "osmotic pressure adjusting agent" is used at a concentration at which the osmotic pressure of the outer aqueous phase is about 1/50 to about 5 times, preferably about 1/25 to about 3 times the osmotic pressure of physiological saline.

As a method for removing the organic solvent, a method known per se or a method analogous thereto is used. For example, a method of evaporating the organic solvent at normal pressure or gradually reducing the pressure while stirring with a propeller-type stirrer or a magnetic stirrer, or a method of evaporating the organic solvent while adjusting the degree of vacuum using a rotary evaporator or the like. And the like. The microcapsules thus obtained are separated by centrifugation or filtration, and the free bioactive substances and emulsifiers attached to the surface of the microcapsules are repeatedly washed with distilled water several times, and then again distilled water and the like. And freeze-dried.

During the above production process, an anti-agglomeration agent may be added to prevent agglomeration of particles. Examples of the "aggregation inhibitor" include, for example, mannitol, lactose, glucose,
Water-soluble polysaccharides such as starches (eg, corn starch), amino acids such as glycine, and proteins such as fibrin and collagen are used. Of these, mannitol is preferred. After freeze-drying, if necessary, under reduced pressure,
Heating may be performed within a condition that the microcapsules do not fuse with each other to remove water and the organic solvent in the microcapsules. Preferably, the polymer is heated at a temperature slightly higher than the midpoint glass transition temperature of the biodegradable polymer determined by a differential scanning calorimeter under the condition of a heating rate of 10 to 20 ° C. per minute.
More preferably, it is heated within a temperature range about 30 ° C. higher than the midpoint glass transition temperature of the biodegradable polymer. For example, when a lactic acid-glycolic acid polymer is used as the biodegradable polymer, a temperature range higher than the midpoint glass transition temperature by 10 ° C. higher than the midpoint glass transition temperature,
Preferably, heating is performed in a temperature range higher than the midpoint glass transition temperature by 5 ° C. than the midpoint glass transition temperature. The heating time varies depending on the amount of the microcapsules, but generally, after the microcapsules themselves reach a predetermined temperature, about 1 hour.
2 hours to about 168 hours, preferably about 24 hours to about 12 hours
0 hours, particularly preferably from about 48 hours to about 96 hours. The heating method is not particularly limited as long as the method is capable of uniformly heating the aggregate of microcapsules.
A method of heating and drying in a fluidized tank, a moving tank or a kiln, a method of heating and drying with a microwave, and the like are used. Among these, the method of heating and drying in a thermostat is preferred.

(II) Phase separation method A coacervation agent is gradually added to an organic solvent solution containing the composition containing the physiologically active substance and the biodegradable polymer described in (I) above with stirring. Microcapsules are precipitated and solidified. The "coacervation agent" is not particularly limited as long as it is a polymer-based, mineral oil-based or vegetable oil-based compound that is miscible with an organic solvent and does not dissolve the biodegradable polymer. Specific examples include silicone oil, sesame oil, soybean oil, corn oil, cottonseed oil, coconut oil, linseed oil, mineral oil, n-hexane,
n-heptane and the like. One or a mixture of two or more of these coacervation agents is used.
The amount of the “coacervation agent” used is about 0.01 to 1,000 times the volume of the oil phase, preferably about 0.05 to 50 times.
It is 0 times, particularly preferably about 0.1 to 200 times. After fractionating the microcapsules thus obtained, (i) washing repeatedly with heptane or the like to remove components (coacervation agent etc.) other than the composition containing the physiologically active substance and the biodegradable polymer, Dried under reduced pressure, or
(Ii) Wash repeatedly with distilled water several times, disperse again in distilled water or the like, freeze-dry, and further heat dry.

(III) Spray drying method An organic solvent solution or dispersion containing the composition containing the physiologically active substance and the biodegradable polymer described in the above (I) is sprayed using a nozzle with a spray drier (spray drier). ), And the organic solvent in the atomized droplets is volatilized within a very short time to prepare microcapsules. Examples of the nozzle include a two-fluid nozzle type, a pressure nozzle type, and a rotating disk type. Thereafter, if necessary, washing may be performed in the same manner as described in the underwater drying method (I), followed by freeze-drying and further heating and drying.

An example of the production method when the composition, especially the sustained-release composition is, for example, fine particles (microparticles) will be described below. An organic solvent solution or dispersion containing the composition containing the physiologically active substance and the biodegradable polymer described in (I) is mixed with an organic solvent and a dispersion while adjusting the degree of vacuum using, for example, a rotary evaporator. After evaporating water to dryness, it is pulverized with a jet mill or the like to obtain fine particles (microparticles). Further, the pulverized fine particles may be washed by the same method as described in the above (I), and then freeze-dried and further dried by heating.

The particle size of the composition of the present invention, particularly the sustained-release composition, may be in a range that satisfies the degree of dispersion and needle penetration when used as a suspension injection, for example.
The average particle size is about 0.1 to 300 μm, preferably about 0.5 to 150 μm, and more preferably about 1 to 100 μm.
μm. The average particle diameter can be measured by a method known per se using, for example, a laser analysis type particle size distribution analyzer (SALD2000A: Shimadzu). The composition of the present invention, particularly the sustained release composition, as it is,
Alternatively, it is formulated into various dosage forms as a raw material and injected or implanted into intramuscular, subcutaneous, organs, etc., transmucosal absorbent into nasal cavity, rectum, uterus, etc., orally [eg, capsule (eg, Hard capsules, soft capsules and the like], solid preparations such as granules and powders, and liquid preparations such as syrups, emulsions and suspensions). Of these, injections are preferred. For example, when the composition of the present invention, in particular, the sustained-release composition is an injection (for injection), the composition is used as a dispersant [eg, a surfactant such as Tween 80, HCO-60, or the like;
Polysaccharides such as sodium hyaluronate, carboxymethylcellulose, sodium alginate, etc.), preservatives (eg, methyl paraben, propyl paraben, etc.), isotonic agents (eg, sodium chloride, mannitol, sorbitol, glucose, proline, etc.) and aqueous It is made into a suspension or dispersed together with vegetable oils such as sesame oil and corn oil to make an injection as an oily suspension. In order to make the composition of the present invention, particularly a sustained-release composition, into a sterile preparation, a method of sterilizing all the production steps, a method of sterilizing with gamma rays,
Examples include a method of adding a preservative, but the method is not particularly limited.

Since the composition of the present invention, particularly the sustained-release composition, has low toxicity, it is safe for mammals (eg, human, cow, pig, dog, cat, mouse, rat, rabbit, etc.). It can be used as a medicine and the like. The composition of the present invention, particularly the sustained release composition, depends on the type of the physiologically active substance contained,
It can be used as a prophylactic / therapeutic agent for various diseases. For example, when the bioactive substance is a GnRH antagonist, for example, sex hormone-dependent cancer (eg, prostate cancer, uterine cancer, breast cancer, pituitary tumor, etc.), bone metastasis of sex hormone-dependent cancer, benign prostatic hyperplasia, uterus Fibroids, endometriosis, precocious puberty, amenorrhea, premenstrual syndrome, multilocular ovary syndrome, acne, baldness, Alzheimer's disease (Alzheimer's disease, Alzheimer's senile dementia and a mixture thereof), etc. Is useful for the prevention and / or treatment of In addition, the compositions of the present invention, especially sustained release compositions, are useful for regulating reproduction in males and females (eg, pregnancy regulators, menstrual cycle regulators, etc.). The compositions of the present invention, especially sustained release compositions, can be further used as contraceptives for men and women, and as ovulation inducers for women. The composition of the present invention, particularly the sustained-release composition, can be used for the treatment of infertility by utilizing the rebound effect after drug withdrawal. In addition, sex hormone-independent
Prevention of H-RH sensitive benign or malignant tumors
It can also be used as a therapeutic. Further, the composition of the present invention, particularly the sustained release composition, is useful in the field of animal husbandry for regulating estrus in animals, improving meat quality for meat, promoting animal growth, and the like. The composition of the present invention, particularly the sustained release composition, is also useful as a spawning promoter for fish.

The compositions of the present invention, especially sustained release compositions, exhibit transient elevations in the blood testosterone (in males) concentration observed upon administration of a GnRH superagonist such as leuprorelin acetate. Phenomenon). The composition of the present invention, in particular, a sustained-release composition, comprises leuprorelin acetate (Leuprorelin), gonadrelin (Gonadrelin), buserelin (tripleline), triptrelin (Goptelilin), and gosereli (Gosereli).
n), nafarelin (Nafarelin), histrelin (Hist
relin), Deslorelin, Meterelin (M
eterelin), GnRH such as Lecirelin
It can be used in combination with a super agonist (preferably leuprorelin acetate). Further, the composition of the present invention, particularly a sustained-release composition, comprises a steroidal or non-steroidal anti-androgen or anti-estrogen, a chemotherapeutic agent, a peptidic GnRH antagonist, a 5α-reductase inhibitor, an α-receptor. Body inhibitor, aromatase inhibitor, 17β
-It is also effective to use in combination with at least one of a hydroxysteroid dehydrogenase inhibitor, an adrenal androgen production inhibitor, a phosphorylase inhibitor, a hormonal therapy agent, a cell growth factor or a drug that inhibits the action of its receptor, etc. is there.

The “chemotherapeutic agent” includes Ifosfamide, UFT, Adriamycin (Adriam
ycin), peplomycin, cisplatin, cyclophosphamide
ide), 5-FU, UFT, methotrexate (Methotrex
ate), mitomycin C, mitoxantrone, taxotere and the like. The “peptidic GnRH antagonist” includes Cetrorelix, Ganirelix
parenterally administered peptide GnRH antagonists such as nirelix) and Abarelix. Examples of the “adrenal androgen production inhibitor” include lyase (C _17,20- lyase) inhibitors and the like. Examples of the "phosphorase inhibitor" include tyrosine phosphorylase. As the "hormone therapy agent",
Examples include antiestrogens, progestin (eg, MPA), androgens, estrogens, antiandrogens, and the like.

The "cell growth factor" may be any substance which promotes cell growth, and is usually a peptide having a molecular weight of 20,000 or less and binding to a receptor. Factors that exert their effects at lower concentrations are mentioned, specifically, (1) EGF (epidermal g
rowth factor) or a substance having substantially the same activity as that (eg, EGF, hallegulin (HER2 ligand))
(2) insulin or a substance having substantially the same activity as insulin (eg, insulin, IGF (insul)
in-like growth factor) -1, IGF-2, etc.),
(3) FGF (fibroblast growth factor) or a substance having substantially the same activity as the FGF (eg, αFGF, β
FGF, KGF (Keratinocyte Growth Factor), HG
F (Hepatocyte Growth Factor), FGF-10, etc.), (4) Other cell growth factors (eg, CSF (colon
y stimulating factor), EPO (erythropoietin),
IL-2 (interleukin-2), NGF (nerve growth fac)
tor), platelet-derived growth factor (PDGF),
TGFβ (transforming growth factor β) and the like. As the “cell growth factor receptor”,
Any receptor may be used as long as it has the ability to bind to the above-mentioned cell growth factor, and specifically, EGF receptor, haregulin receptor (HER2), insulin receptor-1, insulin receptor-2. , IGF receptor, FG
F receptor-1 or FGF receptor-2 and the like. Examples of the drug that inhibits the action of the cell growth factor include:
Herceptin (HER2 receptor antibody) and the like. Examples of the drug that inhibits the action of the cell growth factor or its receptor include herbimycin, PD15
3035 (Science 265 (5175) p1093, (1994)).

[0065] HER2 inhibitors are also examples of agents that inhibit the action of cell growth factors or their receptors.
As a HER2 inhibitor, if it is a substance that inhibits HER2 activity (eg, phosphorylation activity), an antibody, a low molecular compound (synthetic compound, a natural product), an antisense, a HER2 ligand, a hallegulin, or a structure thereof may be used. Any of modified or modified parts may be used. Further, it may be a substance that inhibits HER2 activity by inhibiting HER2 receptor (eg, HER2 receptor antibody). HE
Examples of the low-molecular compound having an R2 inhibitory action include:
Compounds described in WO98 / 03505, specifically 1
-[3- [4- [2-((E) -2-phenylethenyl) -4-oxazolylmethoxy] phenyl] propyl] -1,2,4-triazole and the like. For benign prostatic hyperplasia, GnRH super agonists, antiandrogens, antiestrogens, peptidic GnRH antagonists, 5α-reductase inhibitors, α-receptor inhibitors, aromatase inhibitors, 17β-hydroxysteroid dehydrogenation Combinations of drugs such as enzyme inhibitors, adrenal androgen production inhibitors, and phosphorylase inhibitors with the compositions of the present invention, especially sustained release compositions, may be mentioned.

For prostate cancer, GnRH super agonists,
Anti-androgens, anti-estrogens, chemotherapeutic agents [eg, Ifosfamide, UFT, Adriamycin, Peplomyc
in), cisplatin, etc.], peptidic GnRH antagonists, aromatase inhibitors, 17β-hydroxysteroid dehydrogenase inhibitors, adrenal androgen production inhibitors, phosphorylase inhibitors, hormonal therapeutic agents [eg, estrogens Agents (eg, DSB, EMP, etc.), antiandrogen agents (eg, CMA, etc.), agents that inhibit the action of cell growth factor or its receptor, and the compositions of the present invention, especially sustained release compositions Combination with a product. For breast cancer, GnRH super agonists, antiestrogens, chemotherapeutics [eg, cyclophosphamide (Cycl
ophosphamide), 5-FU, UFT, Methotrexate, Adriamycin,
Mitomycin C, Mitoxantrone, etc.], peptidic GnRH antagonists, aromatase inhibitors, adrenal androgen production inhibitors, phosphorylase inhibitors, hormonal therapies [eg, anti-estrogens [eg , Tamoxifen, etc.), progestin (eg, MPA, etc.), androgen, estrogen, etc.), a drug such as a drug inhibiting the action of cell growth factor or its receptor, and the composition of the present invention. In particular, a combination with a sustained-release composition can be mentioned. The above-mentioned agents may be administered to the same subject at the same time as the composition of the present invention, in particular, a sustained-release composition, or at an interval. In addition, the compositions of the present invention, especially sustained release compositions, can be administered prior to administration of a GnRH superagonist, such as leuprorelin acetate, to provide treatment without flare.

The dosage of the composition of the present invention, particularly the sustained-release composition, varies depending on the type and content of the physiologically active substance, dosage form, duration of release of the physiologically active substance, target disease, target animal, and the like. Any effective amount of a physiologically active substance may be used. The dose of the physiologically active substance per dose may be, for example, when the composition is a one-month preparation, about 0.01 mg to 20 mg / kg body weight, preferably about 0.05 mg to 20 mg / kg body weight per adult (body weight 60 kg). 5 mg / kg body weight. The dose of the composition of the present invention, in particular, the sustained-release composition may be about 0.05 mg to 50 mg per adult (60 kg body weight) per adult.
mg / kg, preferably about 0.1-30 mg / kg
It is. The frequency of administration may be once every several weeks, once a month, or once every several months (eg, three months, four months, six months, etc.), such as the type and content of the physiologically active substance, dosage form, physiological It can be appropriately selected depending on the duration of the active substance release, the target disease, the target animal, and the like. The release period of the physiologically active substance from the composition of the present invention, particularly the sustained-release composition, is not particularly limited because it varies depending on the type of the physiologically active substance, the dosage form of the composition, the dose or the administration site. Time to 1 year, preferably 24 hours to 8 months, more preferably 1 week to
4 months.

The composition of the present invention, particularly the sustained-release composition, has a glass transition point (Tg) of about 10 times as compared with that of a biodegradable polymer having two or more carboxyl groups at a terminal. Those having a high temperature of not less than 0 ° C. are preferred from the viewpoint of stability, more preferably those having a high temperature of not less than about 12 ° C., still more preferably not less than about 15 ° C. Here, the glass transition point refers to a value measured by a differential scanning calorimeter (DSC7, manufactured by Perkin-Elmer). Specifically, using a biodegradable polymer having a glass transition point of about 31 to 42 ° C. (for example, terminal tartronic acid PLA obtained in Reference Example 9 or Reference Example 10 described later), excellent stability is obtained. The composition of the present invention having a glass transition point of about 53 to 58 ° C. can be obtained.

The composition is preferably one that is sustained release. That is, when the non-peptidic physiologically active substance is poorly water-soluble, one that accelerates the release rate of the non-peptidic physiologically active substance is preferable.

[0070]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail with reference to Reference Examples, Examples and Experimental Examples, but these do not limit the present invention.

EXAMPLES Reference Example 1 2-amino-4-methyl-5- (4-nitrophenyl)
Thiophene-3-carboxylic acid ethyl ester 4-nitrophenylacetone (35.0 g, 195 mm
ol), ethyl cyanoacetate (23.8 g, 195 mmol)
l), a mixture of ammonium acetate (3.1 g, 40 mmol) and acetic acid (9.1 ml, 159 mmol)
While removing the water generated by the Dean-Stark apparatus, 24
Heated to reflux for an hour. After cooling, the reaction solution was concentrated under reduced pressure, and the residue was partitioned between dichloromethane and aqueous sodium bicarbonate. The organic layer was washed with brine, dried (MgSO ₄ ), and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography. The obtained oil was dissolved in ethanol, sulfur (5.0 g, 160 mmol) and diethylamine (16.0 ml, 160 mmol) were added, and the mixture was added at 60-70 ° C.
And stirred for 2 hours. After cooling, the reaction solution was concentrated under reduced pressure,
The residue was partitioned between dichloromethane and aqueous sodium bicarbonate. The organic layer was washed with brine, dried (MgSO ₄ ), and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography and crystallized from ether-hexane to give the title compound as red plate crystals (22.2 g, 52%). mp: 168-170 ° C (recrystallized from ether-hexane). ¹ H-NMR (200 MHz, CDCl ₃ ) δ: 1.39
(3H, t, J = 7.1 Hz), 2.40 (3H,
s), 4.34 (2H, q, J = 7.1 Hz), 6.2
7 (2H, br), 7.48 (2H, d, J = 8.7H)
z), 8.23 (2H, d, J = 8.7 Hz). IR (KBr): 3446, 3324, 1667, 15
80, 1545, 1506, 1491, 1475, 14
10,1332 cm ^-1 .

Reference Example 2 5-methyl-6- (4-nitrophenyl) -3-phenylthieno [2,3-d] pyrimidine-2,4 (1H, 3
H) -dione The compound obtained in Reference Example 1 (5.00 g, 16.32 m)
phenylisocyanate (2.66 ml, 24.48 mmol) was added to a pyridine (30 ml) solution, stirred at 45 ° C. for 6 hours, the reaction solution was concentrated under reduced pressure, and the resulting residue was dissolved in ethanol (6 ml). And 28% sodium methoxide (7.86 g, 4
0.80 mmol) was added and the reaction solution was stirred at room temperature for 2 hours, 2N hydrochloric acid (25 ml, 50 mmol) was added, and the ethanol solvent was distilled off under reduced pressure. The obtained residue was filtered, washed with water-ethanol, dried under reduced pressure, and recrystallized from ethanol to give the title compound (6.0) as a yellow powder.
9g, 98%). mp:> 300 ° C. Elemental analysis: C (%) H (%) N (%) calculated as C ₁₉ H ₁₃ N ₃ O ₄ S.0.3 H ₂ O Calculated: 59.30; 3.56; 10.92 Actual: 59. 56; 3.52; 10.93 ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ: 2.
50 (3H, s), 7.31-7.46 (5H, m),
7.78 (2H, d, J = 8.8 Hz), 8.32 (2
H, d, J = 8.8 Hz), 12.50 (1H, s). IR (KBr): 1715, 1657, 1593, 15
10 cm ^-1 .

Reference Example 3 1- (2,6-difluorobenzyl) -5-methyl-6
-(4-nitrophenyl) -3-phenylthieno [2,
3-d] pyrimidine-2,4 (1H, 3H) -dione The compound obtained in Reference Example 2 (52.54 g, 0.131
mol) in dimethylformamide (1.0 l)
Potassium carbonate (19.00 g, 0.138 mol), potassium iodide (22.90 g, 0.138 mol),
2,6-difluorobenzyl chloride (22.40 g,
0.138 mol) and stirred at room temperature for 2 hours.
The residue obtained by concentrating the reaction solution was partitioned between chloroform and brine. The aqueous layer was extracted with chloroform, the combined extracts were washed with brine, dried (MgSO ₄ ), and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography to give the title compound (6) as pale yellow crystals.
1.50 g, 93%). mp: 280-282 [deg.] C. ¹ H-NMR (300 MHz, CDCl ₃ ) δ: 2.57
(3H, s), 5.38 (2H, s), 6.94 (2
H, d, J = 8.1 Hz), 7.42-7.58 (8
H, m), 8.29 (2H, d, J = 8.8 Hz). IR (KBr): 1719, 1669, 1524, 14
73 cm ^-1 .

Reference Example 4 5-bromomethyl-1- (2,6-difluorobenzyl) -6- (4-nitrophenyl) -3-phenylthieno [2,3-d] pyrimidine-2,4 (1H, 3H )-
Dione The compound obtained in Reference Example 3 (30.34 g, 0.060
mol), N-bromosuccinimide (12.81 g,
0.072 mol), a mixture of α, α′-azobisisobutyronitrile (1.15 g, 0.007 mol) and chlorobenzene (450 ml) was stirred at 85 ° C. for 3 hours. After cooling, the reaction solution was washed with brine and dried (MgSO 4).
₄ ) After that, the solvent was distilled off under reduced pressure. The obtained residue was recrystallized from ethyl acetate to give the title compound as yellow needles (80.
21 g, 100%). mp: 228-229 ° C. ¹ H-NMR (300 MHz, CDCl ₃ ) δ: 4.77
(2H, s), 5.38 (2H, s), 6.96 (2
H, t, J = 8.1 Hz), 7.29-7.58 (6
H, m), 7.79 (2H, d, J = 8.5 Hz),
8.35 (2H, d, J = 8.5 Hz). IR (KBr): 1721, 1680, 1524, 14
73,1348 cm ^-1 . FAB-Mass m / z 5
84 (MH) ⁺

Reference Example 5 5- (N-benzyl-N-methylaminomethyl) -1-
(2,6-difluorobenzyl) -6- (4-nitrophenyl) -3-phenylthieno [2,3-d] pyrimidine-2,4 (1H, 3H) -dione The compound obtained in Reference Example 4 ( 80.00g, 0.119
mol) of dimethylformamide (600 ml) under ice-cooling.
ml, 0.155 mol) and benzylmethylamine (18.45 ml, 0.143 mol). After stirring at room temperature for 2 hours, the reaction solution was concentrated, and the obtained residue was partitioned between ethyl acetate and saturated aqueous sodium hydrogen carbonate. The aqueous layer was extracted with ethyl acetate, and the combined organic layers were dried (MgSO ₄ )
Thereafter, the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography to give a yellow oil (74.90 g, 100%), which was recrystallized from ethyl acetate to give the title compound as yellow needles. mp: 173-174 ° C. ¹ H-NMR (300 MHz, CDCl ₃ ) [free amine] δ: 1.31 (3H, s), 3.60 (2H,
s), 3.96 (2H, s), 5.39 (2H, s),
6.95 (2H, t, J = 8.2 Hz), 7.18-
7.55 (11H, m), 8.02 (2H, d, J =
9.0Hz), 8.26 (2H, d, J = 9.0H)
z). IR (KBr) [hydrochloride]: 1719,167
8, 1597, 1520 cm ^-1 .

Reference Example 6 6- (4-aminophenyl) -5- (N-benzyl-N
-Methylaminomethyl) -1- (2,6-difluorobenzyl) -3-phenylthieno [2,3-d] pyrimidine-2,4 (1H, 3H) -dione Compound (3) obtained in Reference Example 5 .00g, 4.80mm
ol) in formic acid (30 ml), under ice cooling, 1M hydrogen chloride-ether (14.4 ml, 14.4 mmol) and 10% palladium on carbon powder (300 mg) were added, and the mixture was stirred with hydrogen at room temperature and pressure for 2 hours. Was added. The reaction solution was filtered through celite, and the residue obtained by concentrating the filtrate under reduced pressure was partitioned between dichloromethane and saturated aqueous sodium hydrogen carbonate. The aqueous layer was extracted with dichloromethane, and the combined organic layers were dried (Mg
After SO ₄ ), the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography to give the title compound as white crystals (2.41 g, 84%). mp: 205-207 ° C. Elemental analysis value C ₃₄ H ₂₈ N ₄ O ₂ SF ₂ 0.1AcOEt
・ As 1.2H ₂ O ¹ H-NMR (300 MHz, CDCl ₃ ) δ: 2.05
(3H, s), 3.56 (2H, s), 3.83 (2
H, br), 3.88 (2H, s), 5.36 (2H,
s), 6.70 (2H, d, J = 8.8 Hz), 6.8
8-6.94 (2H, m), 7.21-7.31 (8
H, m), 7.41-7.53 (5H, m). IR (KBr): 1715, 1657, 1628, 15
37 cm ^-1 .

Reference Example 7 5- (N-benzyl-N-methylaminomethyl) -1-
(2,6-difluorobenzyl) -6- [4- (3-methoxyureido) phenyl] -3-phenylthieno [2,3-d] pyrimidine-2,4 (1H, 3H) -dione In Reference Example 6, The obtained compound (5.0 g, 8.41 mmol)
l) in dichloromethane (120 ml) solution under ice-cooling,
Triethylamine (2.34 ml, 16.82 mmol)
l) was added and stirred. The reaction solution was added with N,
N'-carbonyldiimidazole (2.73 g, 16.
82 mmol), and the mixture was returned to room temperature from under ice-cooling and stirred for 42 hours. Return to ice-cooling again, and add O-methylhydroxylamine hydrochloride (7.02 g, 84.08 mmol).
l) and triethylamine (11.7 ml, 84.0).
8 mmol) was added. The reaction solution was returned from ice cooling to room temperature and stirred for 3 hours. The reaction solution was partitioned between chloroform and saturated aqueous sodium hydrogen carbonate. The aqueous layer was extracted with chloroform, the combined extracts were washed with brine, dried (MgSO ₄ ), and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography to give a pale yellow solid, which was recrystallized from chloroform-ether to give white crystals (4.52).
g, 80%). mp: 204-205 ° C. ¹ H-NMR (300 MHz, CDCl ₃ ) δ: 2.05
(3H, s), 3.57 (2H, s), 3.82 (3
H, s), 3.90 (2H, s), 5.37 (2H,
s), 6.92 (2H, d, J = 8.2 Hz), 7.1
6-7.31 (9H, m), 7.42-7.57 (5
H, m), 7.63 (1H, s), 7.73 (2H,
d, J = 8.8 Hz). IR (KBr): 3338, 3064, 1717, 16
69, 1628, 1591, 1531, 1470c
m ^-1 .

Reference Example 8 3- (N-benzyl-N-methylaminomethyl) -4,
7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4-[(1-hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothieno [2,3-b] pyridine 2 -(4-aminophenyl) -7- (2,6-difluorobenzyl) -4,7-dihydro-5-isobutyryl-
3- (N-benzyl-N-methylaminomethyl) -4-
Oxothieno [2,3-b] pyridine (0.57 g,
1.0 mmol) in dichloromethane (10 ml) solution, diisopropylethylamine (0.52 g, 4 mm
ol) and 2-hydroxycyclopropanecarboxylic acid (0.204 g, 2 mmol), and the mixture was stirred under ice-cooling.
To this solution was added benzotriazol-1-yloxytrisdimethylaminophosphonium hexafluorophosphate (BOP reagent) (1.76 g, 4 mmol). The mixture was stirred for 1 hour under ice cooling, and further stirred at room temperature for 4 days.
The reaction solution was dried under reduced pressure, and the obtained residue was partitioned between water (50 ml) and chloroform (50 ml). The aqueous layer was extracted again with chloroform (10 ml). The combined extracts were washed with brine, dried (MgSO ₄ ), and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography, and recrystallized from ether to obtain yellow powdery crystals (0.27 g, 41%). ¹ H-NMR (300 MHz, CDCl ₃ ) δ: 1.16
-1.20 (2H, m), 1.18 (6H, d), 1.
48-1.51 (2H, m), 2.09 (3H, s),
3.64 (2H, s), 3.95 (1H, br s),
4.14 (2H, s), 4.12-4.19 (1H,
m), 5.20 (2H, s), 6.99 (2H, t),
7.10-7.25 (5H, m), 7.34-7.46
(1H, m), 7.57 (2H, d), 7.70 (2
H, d), 8.21 (1H, s), 8.82 (1H,
s).

Reference Example 9 Dibenzyl tartronate (2.40) cooled to -78 ° C
Under a nitrogen atmosphere, a 1.1 M diethylzinc toluene solution (3.6 ml) was added to g), and the mixture was reacted at room temperature for 20 minutes. To this, DL-lactide (7.04 g) was added and mixed under a nitrogen atmosphere, and polymerized at 130 ° C. for 2 hours. Then, for terminating and deprotecting the polymerization reaction, the reaction product was dissolved in trifluoroacetic acid (10 ml), thioanisole (5.64 ml) was added, and the mixture was stirred at -5 ° C for 1 hour. Methanesulfonic acid (20 ml) was added and the mixture was further stirred for 40 minutes. The reaction solution was cooled with isopropyl ether (1.5
L) and the polymer was precipitated and recovered, and then reprecipitated and purified twice with dichloromethane / cold isopropyl ether. The purified precipitate was dissolved in dichloromethane and washed repeatedly with water until neutral. Next, the dichloromethane solution was concentrated and dried under vacuum (40 ° C., 2 days) to obtain poly (DL-lactic acid) whose ω residue was tartronic acid. ¹ H-NMR
As a result of the analysis, the signal of phenyl hydrogen of the benzyl group had completely disappeared, confirming the deprotection. As a result of the atomic absorption measurement, the residual zinc was detected at the detection limit (10 p.
pm) or less, indicating that the polymerization catalyst was effectively removed by this method. As a result of GPC measurement, the weight average molecular weight was 3,600 and the degree of dispersion was 1.41. Further, when the terminal group labeling quantitative method was applied to this polymer, a strong purple color was exhibited, and regeneration of the carboxyl group by deprotection was confirmed. In addition, from the comparison with the absorbance when tartronic acid hydrazide was used in the ONPH labeling method using tartronic acid as a reference substance, the result of obtaining the amount of tartronic acid, which is the ω residue of the polymer, as the amount of dicarboxyl groups, 378.9 μmol / g.

REFERENCE EXAMPLE 10 Dibenzyl tartronate (2.00
Under a nitrogen atmosphere, a 1.1 M diethylzinc toluene solution (3.0 ml) was added to g), and the mixture was reacted at room temperature for 20 minutes. To this, DL-lactide (19.20 g) was added and mixed under a nitrogen atmosphere, and polymerized at 130 ° C. for 2 hours. Then, for terminating and deprotecting the polymerization reaction, the reaction product was dissolved in trifluoroacetic acid (20 ml), thioanisole (4.70 ml) was added, and the mixture was stirred at -5 ° C for 1 hour. Methanesulfonic acid (20 ml) was added, and the mixture was further stirred for 20 minutes. The reaction solution was cooled with isopropyl ether (1.5
L) and the polymer was precipitated and recovered, and then reprecipitated and purified twice with dichloromethane / cold isopropyl ether. The purified precipitate was dissolved in dichloromethane and washed repeatedly with water until neutral. Next, the dichloromethane solution was concentrated and dried under vacuum (40 ° C., 2 days) to obtain poly (DL-lactic acid) whose ω residue was tartronic acid. ¹ H-NMR
As a result of the analysis, the signal of phenyl hydrogen of the benzyl group had completely disappeared, confirming the deprotection. As a result of the atomic absorption measurement, the residual zinc was detected at the detection limit (10 p.
pm) or less, indicating that the polymerization catalyst was effectively removed by this method. As a result of GPC measurement, the weight average molecular weight was 9,600, and the degree of dispersion was 1.68. Further, when the terminal group labeling quantitative method was applied to this polymer, a strong purple color was exhibited, and regeneration of the carboxyl group by deprotection was confirmed. In addition, from the comparison with the absorbance when tartronic acid hydrazide was used in the ONPH labeling method using tartronic acid as a reference substance, the result of obtaining the amount of tartronic acid, which is the ω residue of the polymer, as the amount of dicarboxyl groups, It was 155.3 μmol / g.

Example 1 PLA terminal tartronic acid obtained in Reference Example 9 (weight average molecular weight = 3,600, number average molecular weight = 2,600)
(2.5 g) was added to and dissolved in a mixture of dichloromethane (14 ml) and methanol (2 ml). After adding the compound obtained in Reference Example 7 (1.4 g) to this solution,
The mixture was mixed and dissolved with a vortex mixer to obtain an organic solvent solution. This organic solvent solution was adjusted to 0.1% (w / v) polyvinyl alcohol (PV
A) (800 ml), and the mixture was converted into an o / w emulsion using a turbine-type homomixer. This o / w type emulsion was stirred at room temperature to evaporate dichloromethane to prepare microcapsules. The obtained microcapsules were separated by centrifugation (about 2,000 rpm). Then, after washing once with distilled water (400 ml), mannitol (450 mg) was added and freeze-dried to obtain powdery microcapsules (3.7 g).

Example 2 PLA terminal tartronic acid obtained in Reference Example 10 (weight average molecular weight = 9,600, number average molecular weight = 5,700)
(2.0 g) was added and dissolved in a mixture of dichloromethane (5 ml) and methanol (0.5 ml). After the compound (1.0 g) obtained in Reference Example 8 was added to this solution, it was mixed and dissolved with a vortex mixer to obtain an organic solvent solution. This organic solvent solution was poured into 0.1% (w / v) polyvinyl alcohol (PVA) (800 ml) which had been previously adjusted to 18 ° C. did. This o
The / w type emulsion was stirred at room temperature to evaporate dichloromethane to prepare microcapsules. The obtained microcapsules were separated by centrifugation (about 3,000 rpm). Next, after washing once with distilled water (400 ml), mannitol (500 mg) was added and freeze-dried to obtain powdery microcapsules (3.0 g).

Experimental Example 1 (1) Microcapsule A Lactic acid-glycolic acid polymer having a terminal monocarboxyl group (lactic acid / glycolic acid = 50/50 (mol%), weight average molecular weight = 14,000) (6.0 g) Was added to and dissolved in a mixture of dichloromethane (20 ml) and methanol (2 ml). After adding the compound (1.9 g) obtained in Reference Example 7 to this solution, it was mixed and dissolved with a vortex mixer to obtain an organic solvent solution. The organic solvent solution was adjusted to 0.1% (w /
v) The mixture was poured into polyvinyl alcohol (PVA) (800 ml), and an o / w emulsion was formed using a turbine homomixer. This o / w type emulsion was stirred at room temperature to evaporate dichloromethane to prepare microcapsules. The obtained microcapsules were separated by centrifugation (about 2,000 rpm). Then, after washing once with distilled water (400 ml), mannitol (4
After adding 50 mg), the mixture was freeze-dried to obtain powdery microcapsules A (4.8 g).

(2) Microcapsule B A lactic acid-glycolic acid polymer having a terminal monocarboxyl group (lactic acid / glycolic acid = 75/25 (mol%), weight average molecular weight = 14,000) (6.5 g) was added to dichloromethane ( 20 ml) and methanol (2 ml). The compound (1.8 g) obtained in Reference Example 7 was added to this solution, and then mixed and dissolved with a vortex mixer to obtain an organic solvent solution. This organic solvent solution is
0.1% (w / v) previously adjusted to 18 ° C
The mixture was poured into polyvinyl alcohol (PVA) (800 ml), and an o / w emulsion was prepared using a turbine-type homomixer. This o / w type emulsion was stirred at room temperature to evaporate dichloromethane to prepare microcapsules. The obtained microcapsules were separated by centrifugation (about 2,000 rpm). Then distilled water (4
After washing once with mannitol (450 ml).
mg) and freeze-dried to obtain powdery microcapsules B (7.1 g). (3) The microcapsules (microcapsules 1), microcapsules A, and microcapsules B obtained in Example 1 were weighed in an amount corresponding to 10 mg of the compound obtained in Reference Example 7, and dispersed in 0.5 ml. Medium (0.2% carboxymethylcellulose, 0.1% polysorbate 80
And 5% mannitol in distilled water).
A 6-week-old male SD rat was dosed subcutaneously on the back with a 22G needle. After a predetermined time from the administration, the rat was sacrificed, and the microcapsules remaining at the administration site were taken out. The compound obtained in Reference Example 7 was quantified and divided by its initial content to obtain a residual ratio [Table 1].

[Table 1] Microcapsule 1 Microcapsule A Microcapsule B 101.2% 100.7% 96.9% 1 week 91.9% 86.9% 89.9% 2 weeks 71.9% 85.8% 83.3 % 3 weeks 54.8% 80.7% 77.1% 4 weeks 52.0% 79.7% 70.7% In the results of Table 1, a lactic acid-glycolic acid polymer having a terminal monocarboxyl group was used. From the microcapsules A and B prepared as described above, the release of the compound obtained in Reference Example 7, which is poorly water-soluble, is remarkably slow, and is independent of the lactic acid / glycolic acid ratio which usually affects the release rate. It was shown that release control was difficult. on the other hand,
From the microcapsules 1 of the present invention, it is clear that the release of the compound obtained in Reference Example 7, which is poorly water-soluble, is accelerated.

Experimental Example 2 The glass transition points (Tg) of Microcapsule 1, Microcapsule A and Microcapsule B were measured with a differential scanning calorimeter (DSC7, manufactured by Perkin-Elmer). 0.0 ° C, microcapsule A: 43.7 ° C, microcapsule B: 45.4
° C. The microcapsules 1 of the present invention having a weight-average molecular weight of 3,600 exhibited a high Tg such that the terminal could not be predicted by a monocarboxyl group. Since a high Tg is advantageous for the stability of the preparation, it can be seen that the composition of the present invention is excellent in stability.

Experimental Example 3 (1) Microcapsule C Lactic acid-glycolic acid polymer having a terminal monocarboxyl group (lactic acid / glycolic acid = 75/25 (mol%), weight average molecular weight = 12,000) (2.0 g) Was added to and dissolved in a mixture of dichloromethane (5 ml) and methanol (0.5 ml). After the compound (1.0 g) obtained in Reference Example 8 was added to this solution, it was mixed and dissolved with a vortex mixer to obtain an organic solvent solution. The organic solvent solution was adjusted to 0.1% (w /
v) The mixture was poured into polyvinyl alcohol (PVA) (800 ml), and an o / w emulsion was formed using a turbine homomixer. This o / w type emulsion was stirred at room temperature to evaporate dichloromethane to prepare microcapsules. The obtained microcapsules were separated by centrifugation (about 2,000 rpm). Then, after washing once with distilled water (400 ml), mannitol (5
Then, the mixture was freeze-dried to obtain powdery microcapsules C (3.0 g). (2) The microcapsules (microcapsules 2) obtained in Example 2 and the microcapsules C were weighed in an amount equivalent to 10 mg of the compound obtained in Reference Example 8,
Dispersed in 0.5 ml of a dispersion medium (distilled water in which 0.2% carboxymethylcellulose, 0.1% polysorbate 80 and 5% mannitol are dissolved), and administered subcutaneously to the back of a 6-week-old male SD rat with a 22G injection needle. did. At a predetermined time after the administration, three rats were sacrificed in each group, and the subcutaneous irritation of the microcapsules was visually observed and recorded. [Table 2] shows the number of rats with three subcutaneous irritation after microcapsule administration. The results in Table 2 show that microcapsules C prepared using a lactic acid-glycolic acid polymer having a terminal monocarboxyl group have difficulty in suppressing the subcutaneous irritation of the compound obtained in Reference Example 8. Was. On the other hand, it was revealed that the microcapsules 2 of the present invention completely suppressed the subcutaneous irritation of the compound obtained in Reference Example 8.

[0087]

The composition of the present invention can exhibit a high pharmacological effect by increasing the content of the non-peptide physiologically active substance and controlling or accelerating the release thereof. In addition, in the case where the non-peptide physiologically active substance has subcutaneous irritation derived from a basic substance, the composition of the present invention is expected to have an effect of eliminating the irritation by a strong acidic group at the terminal. Furthermore, the compositions according to the invention have a high glass transition point and are very stable.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 47/34 A61K 47/34 A61P 5/06 A61P 5/06 C07D 495/04 105 C07D 495/04 105A 105Z F-term (reference) 4C071 AA01 BB01 CC01 CC02 CC21 EE12 FF05 FF06 FF10 GG01 GG05 HH08 JJ01 LL01 4C076 AA11 AA61 AA94 BB11 CC30 EE24 FF31 FF33 4C084 AA27 DB09 DB10 DB25 DB71 4C012 MA29 MA04

Claims (21)

[Claims] 1. A non-peptidic physiologically active substance and a terminal 2
A composition comprising a biodegradable polymer having at least two carboxyl groups or a salt thereof. 2. A non-peptidic physiologically active substance and 2
A composition comprising a biodegradable polymer having at least two carboxyl groups or a salt thereof. 3. The composition has a glass transition point (Tg) of 2 at the terminal.
3. The composition according to claim 2, wherein the temperature is about 10 [deg.] C. or higher than the glass transition point of the biodegradable polymer having at least one carboxyl group. 4. The biodegradable polymer having two or more carboxyl groups at a terminal is a polymer having an α, α-dicarboxyl group or an α, β, β′-tricarboxyl group at a terminal. Or the composition according to 2. 5. A biodegradable polymer having two or more carboxyl groups at a terminal, a polyα-hydroxycarboxylic acid having an α, α-dicarboxyl group or an α, β, β′-tricarboxyl group at a terminal. Claim 1 or 2 which is
A composition as described. 6. The composition according to claim 5, wherein the polyα-hydroxycarboxylic acid is linear. 7. A biodegradable polymer having two or more carboxyl groups at a terminal, a lactic acid-glycolic acid polymer having an α, α-dicarboxyl group or an α, β, β′-tricarboxyl group at a terminal The composition according to claim 1, wherein the composition is: 8. The composition according to claim 1, wherein the biodegradable polymer having two or more carboxyl groups at the terminal is a lactic acid-glycolic acid polymer whose ω residue is tartronic acid or citric acid. . 9. The composition according to claim 1, wherein the biodegradable polymer having two or more carboxyl groups at the terminal is polylactic acid whose ω residue is tartronic acid or citric acid. 10. The composition according to claim 1, wherein the non-peptidic physiologically active substance is poorly water-soluble. 11. The composition according to claim 10, which is sustained release. 12. The non-peptide physiologically active substance having a molecular weight of:
12. The composition of claim 11, which is less than about 1,000. 13. The composition according to claim 11, wherein the non-peptide physiologically active substance is a gonadotropin releasing hormone agonist or antagonist. 14. The non-peptidic physiologically active substance has the formula: Wherein, X is a carbon atom or a nitrogen atom, - - - indicates a single bond or a double bond The compound or composition of claim 11, wherein a salt thereof having a partial structure represented by. 15. A compound of the formula Wherein, X is a carbon atom or a nitrogen atom, - - - indicates a single bond or a double bond], or a salt thereof having a partial structure represented by the formula ## STR3 ## Wherein R ¹ and R ² each represent a hydrogen atom, a hydroxy group, a C _1-4 alkoxy group, a C _1-4 alkoxy-carbonyl group or a C _1-4 alkyl group which may have a substituent. , R ³ represents a hydrogen atom, a halogen atom, a hydroxy group or a C _1-4 alkoxy group which may have a substituent, or two adjacent R ³ are linked to form a C _1-4 alkylenedioxy. R ⁴ represents a hydrogen atom or a C _1-4 alkyl group, and R ⁶ represents a C 5 optionally substituted group.
_1-4 alkyl group or formula (Wherein R ⁵ represents a hydrogen atom or R ⁴ and R ⁵ may be linked to form a heterocyclic ring), and n represents an integer of 0 to 5 15. The composition according to claim 14, which is a compound represented by the formula: or a salt thereof. 16. A compound of the formula Wherein, X is a carbon atom or a nitrogen atom, - - - indicates a single bond or a double bond], or a salt thereof having a partial structure represented by the, 5-(N-benzyl -N- methyl Aminomethyl) -1- (2,6-difluorobenzyl) -6- [4- (3-methoxyureido) phenyl]
-3-phenylthieno [2,3-d] pyrimidine-2,
4. The compound according to claim 1, which is 4 (1H, 3H) -dione or a salt thereof.
4. The composition according to 4. 17. A compound of the formulaWherein X represents a carbon atom or a nitrogen atom,---Is simple
Or a double bond).
The compound or a salt thereof has the formula:[Wherein, R⁹Is optionally substituted C_1-7Alkyl group,
Optionally substituted C _3-7Cycloalkyl group, substituted
C that may be_1-6Alkoxyamino group or substituted
An optionally substituted hydroxyamino group is represented by R^TenIs replaced
May be C_1-7Alkyl group or substituted
Each represents a good phenyl group;⁹Is unsubstituted C_1-7A
When it is a alkyl group, R^TenIs the substituted C_1-7Alkyl
Represents a group or substituted phenyl]
Or the salt thereof. 18. A compound of the formula Wherein, X is a carbon atom or a nitrogen atom, - - - indicates a single bond or a double bond], or a salt thereof having a partial structure represented by the, 3- (N-benzyl -N- methyl Aminomethyl) -4,7-dihydro-5-isobutyryl-7- (2,6-difluorobenzyl) -2- [4-
The composition according to claim 14, which is [(1-hydroxycyclopropyl) carbonylamino] phenyl] -4-oxothieno [2,3-b] pyridine or a salt thereof. 19. The composition according to claim 1, which is for injection. 20. A sustained release microcapsule.
Or the composition according to 2. 21. A method for producing a composition, which comprises mixing a non-peptidic physiologically active substance and a biodegradable polymer having two or more carboxyl groups at a terminal or a salt thereof.