CA2130868A1

CA2130868A1 - Sustained releasable parenteral pharmaceutical preparations and method of producing the same

Info

Publication number: CA2130868A1
Application number: CA002130868A
Authority: CA
Inventors: Hiroaki Okada; Yutaka Yamagata; Katsumi Iga
Original assignee: Takeda Chemical Industries Ltd
Current assignee: Takeda Pharmaceutical Co Ltd
Priority date: 1993-08-26
Filing date: 1994-08-25
Publication date: 1995-02-27
Also published as: ATE148625T1; DE69401691D1; KR950005329A; CN1105557C; TW282403B; EP0640336B1; DK0640336T3; US5628993A; KR100338400B1; JPH07112940A; EP0640336A1; DE69401691T2; CN1104488A; US5750100A

Abstract

ABSTRACT

A parenteral pharmaceutical preparation comprises a matrix containing a physiologically active peptide or protein and a polyglycerol diester of a saturated fatty acid, and the matrix is in a solid form at room temper-ature. The molecular weight of the physiologically active peptide or protein is 2,000 dalton or more. The saturated fatty acid includes fatty acids having about 16 to 30 carbon atoms such as palmitic acid, stearic acid, etc. The matrix may be in a pillar or granular form. The parenteral pharmaceutical preparation can be used as an injectable solid administered subcutaneously or intramuscularly (for example, a pellet or tablet for implantation), a suppository or the like, and can release the physiologically active peptide or protein sustainedly for a prolonged period of one week or more.

Description

^' 21~0~68 SUSTAINED RELEASABLE PARENTERAL PHARMACEUTICAL
PREPARATIONS AND METHOD OF PRODUCING THE SAME

FIELD OF THE INVENTION
The present invention relates to a sustained -~
releasable parenteral preparation useful for sustained ~i or prolonged release of a physiologically active pep-tide or protein, and a method of producing th same.

BACKGROUND OF THE INVENTION
For administration of a therapeutic agent, oral ~;-administration is generally employed. Oral administra-tion of a physiologically active peptide or protein, ~-however, causes hydrolysis of the peptide or protein by a digestive enzyme to decrease disadvantageously the absorbability from the digestive tract. Accordingly, such physiologically active peptide or protein is usually administered by repetition of intramuscular or ;
subcutaneous injections or by intravenous drip infu-sion. These methods, however, are not preferable in a chronic administration, although they are acceptable in ! ' a case where the repetition of the injection is ex-tremely limited. By way of illustrating, for the therapy of viral hepatitis type C, interferon-a is 25 continuously administered daily throughout 4 weeks or ;
more (see "Journal of Clinical and Experimental Medi-cine (IGAKU NO AYUMI)", 161, 5, 359-363 (1992)). In ,~, ^~ ~131~86~

such chronic or frequent administration, however, the patient is obliged to be restrained to a great extent.
Therefore, development of an effective and economic administration system for such physiologically active S peptide or protein has been demanded.
Japanese Patent Application Laid-open No.
2930/1988 (JP-A-63-2930) discloses a system where a polypeptide is dispersed in a polylactide. Japanese Patent Publication No. 502117/1988 (JP-B-63-502117) and Japanese Patent Application Laid-open No. 234820/1992 (JP-A-4-234820) disclose pharmaceutical preparations uslng a liposome, and Japanese Patent Publication No.
502574/1991 (JP-B-3-502574) proposes a pharmaceutical preparation where a liposome containing a physiologi-cally active polypeptide is dispersed in a gel.
When these pharmaceutical preparations are admin-istered, however, the drugs are unexpectedly released to a large extent in the initial stage of administra-tion. Thus the drug concentration in blood is in-creaged and the releasing rate of the drug can not be maintained in a certain range. Furthermore, since an - organic solvent is used in the manufacture of the preparation, the polypeptide is denaturated to decrease the physiological activity.
Japanese Patent Application Laid-open No.
2930/1988 (JP-A-63-2930) discloses a sustained releas-able system for a physiologically active polypeptide, `` ~130~8 ,:

which comprises an atherocollagen-matrix and the physi-ologically active polypeptide dispersed in the matrix.
The atherocollagen used as a base is, however, derived from a foreign or different animal from human being, and it may probably show antigenicity.
Japanese Patent Application Laid-open No.
22012/1988 (JP-A-63-22012) discloses a system prepared by dispersing a physiologically active polypeptide in a water-insoluble matrix and compression-molding the dispersion, as a sustained releasable pharmaceutical preparation for parenteral administration of the poly-peptide. The pharmaceutical preparation controls the release of the physiologically active polypeptide by utilizing erosion of the matrix in Yivo. Therefore, the polypeptide may be enzymatically degraded or decom-posed so as to lower the biological availability.
Japanese Patent Application Laid-open No.
85328~1986 (JP-A-61-85328) discloses a pharmaceutical preparation which comprises a composition of a physio-logically active polypeptide and a polyglycerol fattyacid ester, wherein the polyglycerol fatty acid ester is dispersed in water. The pharmaceutical preparation is, however, restricted with regard to a dosage form since it is a solution. Furthermore, since the polyg-lycerol fatty acid ester is utilized to promote thepercutaneous absorption of the physiologically active polypeptide, and the drug can hardly be released for a ~ -longer period of time, e.g. for 24 hours or more.
SUMMARY OF THE INVENTION
Accordingly, it is attempted in the present invention to provide a sustained releasable parenteral pharmaceutical preparation which is free from one or mcre disadvantages associated with the prior art.
It is also attempted in the present invention to provide a method of producing a pharmaceutical preparation, by which the pharmaceutical preparation can be produced in a simple and easy manner.
After intensive investigation and research, the inventors of the present invention found that a pharmaceutical preparation obtainable by use of specific polyglycerol fatty acid esters selected from numerous polyglycerol fatty acid esters in combination with a physiologically active polypeptide or protein can remarkably improve the sustained release of the physiologically active peptide or protein, and can release the physiologically active peptide or protein sustainedly for a prolonged period. The present invention has been accomplished based on these findings.
Thus, the sustained releasable parenteral pharma-ceutical preparation of the present invention comprises a matrix comprising an effective amount of a physiologically active peptide or protein (herelnafter, as far as not partlcularly `

2 1 3 0 g 6 8 28279-17 mentioned, referred to simply as the physiologically active polypeptide) and a polyglycerol diester of a saturated fatty acid. The average molecular weight of the physiologically active polypeptide may frequently ,., ;.' '' ' .,: ~,- ~

~ 213~8~

be 2,000 dalton or more, and the diester may be, in many cases, a diester formed with a polyglycerol having an average polymerization degree of about 4 and a saturated fatty acid having 16 to 30 carbon atoms. The matrix may be in a pillar, granular or other form. The matrix may be an injectable solid for implantation.
The sustained releasable pharmaceutical prepara-tion may be prepared by mixing a physiologically active peptide or protein with a molten or softened polygly-cerol diester of a saturated fatty acid and molding themolten mixture.

DETAILED DESCRIPTION OF THE INVENTION
In this specification, the term "polyglycerol having an average polymerization degree of 4" means a polymerization degree of polyglycerol as a main com-pound which is estimated by the terminal analysis of hydroxyl value, and includes tetraglycerol, as well as a mixture of tetraglycerol as a main component and an unavoidable glycerol or glycerol polymer (for example, diglycerol, triglycerol, pentaglycerol, etc.). There-fore, the average polymerization degree of the polygly-cerol may be about 3.7 to 4.3.
The term "dlester" means the average value of the ester bonds of a main compound which is estimated from the ester value of the polyglycerol fatty acid ester, and includes not only a diester but also a mixture ' ... . ..
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comprising a diester as a main component, and a mono-ester, a triester which may be coexistent or contamina-tive unavoidably. Thus, the average value of the ester bond in the diester may be about 1.7 to 2.3.
In cases where the matrix or the polyglycerol diester of a fatty acid is not a single compound but a mixture, the substance does not show a distinct melting point but softens at a specific temperature. The term "melting point" as used in this specification includes, within the meaning thereof, the softening point of such a mlxture as well.
As the physiologically active polypeptide in the present invention, various peptides and proteins having physiological activities can be used. The average molecular weight of the physiologically active polypep-tide is, for example, about 2,000 dalton or more, preferably about 5,000 to 1,000,000 dalton, more pref- -erably about 10,000 to 500,000 dalton and particularly about 10,000 to 100,000 dalton. Preferred physiologi~
cally active polypeptide includes molecules classified into proteins which is expressed as having a higher-dimensional structure in the field of biochemistry.
The physiologically active polypeptide includes classificatorily, for example, protelns, enzymes/ nu-cleoproteins, glycoproteins, lipoproteins, polypeptideshaving a hormone-like activity, agonists of these - molecules, synthetic analogues including a~tagonists ~ "

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and so on.
The present invention may be applied to a variety of physiologically active polypeptides and the species thereof is not critically restricted. As examples of the physiologically active polypeptides, there may be mentioned immune-controlling factors, lymphokines, monokines, cytokines, enzymes, antibodies, growth stimulating factors, growth suppressing factors, hor-mones, vaccines (including antigens of viruses, bacte-ria, parasites and rickettsiae), blood coagulating fac-tors, and various precursor proteins thereof, mutant proteins, and other su~stances analogous thereto.
To be specified, the physiologically active poly-peptide includes, for example, the following physiolog-ically active high molecular compounds, mutant proteinsand analogues thereto.
(1) Interferons (a-, ~ , etc.), interleukins (IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11), antiallergic factors, suppressor fac-tors, cytotoxic glycoproteins, immuno-cytotoxic fac-tors, immuno-toxins, lymphotoxins, tumor necrosis factors (TNF-a, TNF-~, or the like), cachectin, oncos-tatins, transforming growth factors (TGF-a, TGF-~ and so on), hemopoietic factors (for example, erythropoie-tin), granulocyte-colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), macrophage-colony stimulating factor (M-CSF), ~ . . . . .

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g macrophage peptides, B-cell factors (e.g. B-cell growth factor, etc.), T-cell factors and so on.
(2) Growth factors, for instance, nerve growth factor (NGF), nerve trophic factor (NTF), polypeptides having actions on cranial nerve cells, epitheliocyte growth factor (EGF), insulin-like growth factor (IGF), growth hormone (GH), fibroblast growth factor (FGF), osteogen growth factor, atrical natriuretic factor (ANP), cartilage inducing factor and others. The physiologically active polypeptide belonging to this category further includes, for instance, parathyroid hormone (PTH), endoserine and the like.

(3) Physiologically active polypeptides having a platelet growing action such as platelet-derived growth factor (PDGF), etc.

(4) Physiologically active polypeptides having an enzymatlc action including, for instance, factor VIII, factor IX, fibrinolysis factor, tissue plasminogen activator (TPA), urokinase, prourokinase, streptoki-nase, lipocortin, macrocortin, protein C, C-reactive protein, renin-inhibitor, metalloproteases, tissue inhibitor of metalloprotease (TIMP), superoxide dismu-tase (SOD) and so on.

(5) Physiologically active polypeptides having a hormone-like action such as insulin, secretin, growth hormone releasing factor (GRF), glucagon, gastrins, prolactin, adrenocorticotropic hormone (ACTH), thyroid-stimulating hormone (TSH), luteinizing hormone (LH), follicle-stimulating hormone (FSH), cholecystokinin, human chorionic gonadotropin (HCG), leukokinin, thymo-cin, motilin, kallikrein, etc.

(6) Physiologically active polypeptides acting as a vaccine antigen including antigens such as HTLV-I, HTLV-II, AIDS virus group (e.g. HTLV-III~LAV/HIV and HIV-2, etc.), cytomegalovirus, hepatitis A virus, hepatitis B virus, hepatitis C virus, herpes simplex-I
virus, herpes simplex-II virus, malaria, larvate rabies retrovirus, infectious gastroenteritis virus, parain-fluenza virus, influenza virus, rotavirus group, respi-ratory syncytial virus, varicella-zoster virus, Ep-stein-Barr virus, pertussis; and Gram negative bacteria such as Pseudomonas, endotoxins, tetanus toxin, and others. Such physiologically active polypeptide may be administered singly, or bonded with a hapten, or in combination with an adjuvant.
These physiologically active polypeptide may be naturally-occurring, or be prepared by genetic recombi-nation. The physiologically active polypeptide may have a glycosyl chain, and the structure of glycosyl chaln may be different. Further, examples of the -~
physiologically active polypeptide include mutants, derivatives, relatives or analogues, or active frag-ments of the peptides and proteins mentioned above.
The physiologically active polypeptide may be used . . " ~ ", , ,~, " . . i .,. ,:, , :: ;, . ~ "

~ 213~68 singly or in combination. A substance which can acti-vate the physiologically active polypeptide and/or other ingredient having arithmetic or synergistic effects with the substance can advantageously be em-ployed in combination with the physiologically activepolypeptide. For instance, interferon-~ can be used in combination with the activating substance or ingredient such as interleukins, lentinan, minophagen or the like.
The activating substance or ingredient may be used ~-singly or in combination, with the physiologically active polypeptide.
The present invention is characterized in that the physiologically active polypeptide is sustainedly releasable for a prolonged period by means of combining specific polyglycerol fatty acid esters among a great number of polyglycerol fatty acid esters.
In the polyglycerol fatty acid ester, the term "polyglycerol" means "a polyhydric alcohol having in each molecule thereof n (when cyclic) to n+2 (when stralght-chained or branched) hydroxyl groups, and n-1 ~when straight-chained or branched) to n (when cyclic) ! I ether bonds" ["Polyglycerol Ester" edited and published by Sakamoto Yakuhin Kogyo Co., Ltd., Japan; pp 12, (May 2, 1986)]. Polyglycerol can be obtalned by dehydrating condensation of glycerol, or recovery from residue of glycerol distillation.
As the component used in combination with the 8 6 ~

physiologically active polypeptide, a polyglycerol diester of a saturated fatty acid is employed in the present invention. When the fatty acid is an unsatu-rated fatty acid, the physiologically active polypep-tide may be released in an early stage, and the sus-tained release may be extremely decreased or reduced.
Furthermore, even when the compound is an ester formed with a polyglycerol and a saturated fatty acid, a monoester, a triester, a tetraester, a pentaester, or the like may harm or reduce the sustained release of the physiologically active polypeptide remarkably.
Examples of the saturated fatty acid include a saturated fatty acid having 16 to 30 carbon atoms such as palmitic acid, datulic acid, stearic acid, arachic acid, behenic acid, lignoceric acid, cerotic acid, montanic acid, melissic acid and so on. Preferred examples of the saturated fatty acid include a fatty acid having 16 to 22 carbon atoms (for example, palmit-ic acid, stearic acid, behenic acid, etc.), particular-ly palmitic acid, stearic acid and the like.
The diesters of these saturated fatty acids may bediester formed with a single fatty acid or with a mix-ture of two or more of fatty acids.
The polymerization degree of the polyglycerol is not particularly limited to a specific value which does not adversely affect on the sustained releasing proper-ties, and is selected from the range depending on the ' i : '- ~ '` ., `. ' :. , ' . . .' ' :. ! . '., . ' . .~

~ ~13~6~
, , species of the fatty acid.
Preferred polyglycerol has an average polymeriza-. , .
tion degree of about 3 to 5, particularly 4. Where the average polymerization degree is less than 3 or more than S, depending on the species of the saturated fatty acid, the physiologically active polypeptide may be apt to be released or liberated readily, and, in some cases, may hardly be imparted with high sustained releasing properties. Therefore, a polyglycerol having an average polymerization degree of 4 is advisably employed.
The melting point of the polyglycerol diester of the saturated fatty acid is for example about 40 to 60-C, preferably about 42 to 58-C and more preferably about 45 to 55'C.
The preferred polyglycerol diester of the saturat-ed fatty acid i9 formed with a polyglycerol having an average polymerization degree of 4 and a saturated fatty acid having 16 to 22 carbon atoms.
Examples of the polyglycerol diester having of the saturated fatty acid includes triglycerol diesters such 'a~ triglycerol dipalmitate, triglycerol distearate and triglycerol dibehenate; tetraglycerol diesters such as tetraglycerol dipalmitate, tetraglycerol didatulate, tetraglycerol distearate, tetraglycerol diarachate, tetraglycerol dibehenate, tetraglycerol dilignocerate, tetraglycerol dicerotate, tetraglycerol dimontante, - .
- :: ", . . : .~ , : , :

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tetraglycerol dimelissate, tetraglycerol monopalmitate monostearate, tetraglycerol monopalmitate monobehenate and tetraglycerol monostearate monobehenate; pentagly-cerol diesters such as pentaglycerol dipalmitate, pentaglycerol distearate, pentaglycerol dibehenate and the like. Preferred examples of the diester include tetraglycerol diesters such as tetraglycerol dipalmi-tate, tetraglycerol distearate, tetraglycerol dibehe-nate and others. These diesters can be employed inde-10 pendently or in combination. -The polyglycerol diester is usable for an emulsi-fier as a food additive, and the safety in vivo has already been confirmed. Further, the diester may finally be absorbed ~n v~vo and moreover present no antigenicity.
Matrixes comprising a polyglycerol higher fatty acid ester and a peptide or protein are disclosed in Japanese Patent Application ~aid-open No. 223533/1990 (JP-A-2-223533), EP-A 443572, Japanese Patent Applica-- 20 tion Laid-open Nos. 237/1993 (JP-A-5-2237) and 132416/1993 (JP-A-5-132416). However, these prior literatures do not disclose that a matrix comprising a combinatlon of a polyglycerol diester of the saturated fatty acid and a physiologically active polypeptide can release the physiologically active polypeptide for an extremely prolonged time period.

The pharmaceutical preparation of the present ~

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21~0868 invention is composed of a matrix comprising the physiologically active polypeptide and the diester. Usually, preferred preparation is formed with a matrix wherein the physiologically -active polypeptide is dispersed in the diester. The matrix is preferably in a solid form at room temperature or ambient temperature (5C to 35C), and, usually, the physiologically active polypeptide is homogeneously dispersed in the diester.
When the physiologically active polypeptide is dissolved in the ;
molten diester in the manufacturing procedure, it is preferable to mix homogeneously to give a pharmaceutical preparation where the polypeptide is dispersed in a solid form at room temperature.
Such pharmaceutical preparation is characterized by suppressing the release of the physiologically active polypeptide in the early stage of the administration to a great extent, and sustainedly or continuously releasing of the peptide. for a prolonged time period with maintaining the higher-dimensional structure thereof. `
The ratio of the physiologically active polypeptide relatlve to the diester can be selected from a wide range, and the proportion of the physiologically active polypeptide in the matrix composed of the two components is, for instance, about 0.0001 to 50% by weight, preferably about 0.001 to 20% by weight and more preferably about 0.01 to 10% by weight, and the residue is formed with the diester. In a specific preferred embodiment, the physiologically actlve polypeptide may be from about 0. 1 to 5% by weight based on the polyglycerol diester.

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2~g68 - 15a -For a practical use, the pharmaceutical preparation may be put in a commercial package. Such a commercial package often carries instructions or indications that the pharmaceutical -.
preparation is a sustained release parenteral preparation. ~-,.,,;.~

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~ ~-The matrix may be added, if required, with an ingredient commonly used in the field of solid pharma-ceutical preparations such as an excipient, a binder and a disintegration agent, as well as various addi-tives such as a stabilizing agent, a preservative andthe like. Examples of the stabilizing agent include gelatin, albumin, globulin, protamine, trehalose, D-glucose, dextran and others. As the preservative, there may be mentioned, for instance, paraoxybenzoic acid esters (for example, methylparaben, propylparaben, etc.), benzyl alcohol, chlorobutanol, thimerosal and so on.
The sustained releasable parenteral pharmaceutical preparation may be in any form so far as to be adminis-tered parenterally or non-brally, and is, usually, formed with a matrix in such a dosage form that will not give a patient an excessive pain or suffering, for example, a small or compact matrix. A characteristic of the present invention is that even such small or compact matrix as to be administrable by means of, for instance, a needle for injection, the physiologically active polypeptide can be released sustainedly for a prolonged period. By way of illustration, non-oral adminlstratlon of the present pharmaceutical prépara-tion can prolong the period of the physiologicallyactive polypeptide in blood, for instance, 7 days or more, in comparison with a single or separate adminis-~13U86~

tration of the physiologically active polypeptide.
Thus, the dosage time of the preparation and pain or suffering given to the patient can extremely be re-duced. -~
The pharmaceutical preparation may be utilized as, for instance, an injectable solid which is administra-ble subcutaneously or intramuscularly (e.g. a pellet or implant, etc.), or a transmucosally absorbable composi-tion such as a suppository. The shape or form of the preparation can be selected from a range depending on the dosage form, and may, for instance, be in a powdery or granular form as a powder, a granule or a pill; in a flat, ellipse, rod or pillar form as an injectable pellet or tablet for implantation; or in a spherical or oval form as a suppository. When used as an injection, the preparation may frequently be in a pillar or pow-dery form. Preferred form of the pharmaceutical prepa-ration includes, for instance, pillar form such as cylindrical or columnar form and granular form such as 8pherical form.
The size of the parenteral or non-oral pharmaceu-tical preparation of the present invention may also be 8elected according to the dosage form, as far as it will not pain a patient to an excessive extent. For an injection, when the preparation is a pillar-formed ~atrix, the size is for example about 3 mm or less in diameter and about 30 mm or less in length, preferably .
- - ~130868 .. ~ .

about 1 mm or less in diameter and about 20 mm or less in length which can be administered by using a needle of 11 G or less, more preferably about 0.1 to 1 mm in diameter and about 1 to 20 mm in length, and practical-ly preferred is in a cylindrical or columnar form. Thegrain or particle size of an injectable granular matrix is, in maximum diameter, about 1 mm or less, preferably about 150 ~m or less and more preferably about 1 to 100 ~m. The weight of the matrix may be chosen depending on the form or shape of th pharmaceutical preparation, and ls usually, for example, about gO mg or less and preferably about 1 to 25 mg for an injection.
The pharmaceutical preparation of the present invention can be prepared by various methods, and the preferred is such that using no organic solvent which denatures the polypeptide. As such a method, there may be mentioned, for instance, a process which comprises mixing a physiologically active polypeptide to a molten or softened polyglycerol diester of the saturated fatty acid, and molding the resultant molten mixture into a preparation. Although the physiologically active polypeptide is thermodynamically unstable in an aqueous solutlon, it i8 unexpectedly stable in a solid form such as a freeze-dried powder. Therefore, the physio-logically active polypeptide in a solid powdery orgranular form such as a dried powder is preferably used for homogeneous mixing.

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In the molding, any molding method can be employed according to the form or shape of the pharmaceutical preparation. For instance, an injection can be pre-pared by sucking up the molten mixture into a syringe with a needle and extruding the charged from the needle to give a pillar product or by dropping the molten mixture onto a rotary plate or disk and centrifuging ~-or tumbling the droplets to obtain a spherical product. ~
Further, a fine particulate pharmaceutical preparation ~-10 can be produced by atomizing or spraying the molten ~ -mixture and chilling the powdery product, or by sub-~ectlng the shaped product such as a pellet to a pul-verizing means isuch as a jet mill to obtain a fine particle.
The following examples and experimental example ;
are merely intended to illustrate the present invention in further detail and should not be construed as defin-ing the scope of the invention.

EXAMPLES
Example 1 Tetraglycerol dipalmitate (300 mg: the number of ester bond: 2.0; manufactured by Sakamoto Yakuhin Kogyo Co., Ltd., Japan) was heated at 48'C for melting, and was added with 7.2 mg of a freeze-dried powdery inter-feron-a. A part of the molten mixture was sucked up into a needle of 11 G by use of a syringe, cooled at 21~0868 room temperature and the charged was extruded from the needle to give a cylindrical matrix pharmaceutical preparation (1 mm in diameter, 10 mm in length, about 10 mg in weight).
Example 2 Tetraglycerol distearate (300 mg; the number of -ester bonds: 2.0; manufactured by Sakamoto Yakuhin Kogyo Co., Ltd., Japan) was heated at 58 C for melting, and to the molten was added 7.2 mg of a freeze-dried powder of interferon-a. A portion of the molten mix-ture was sucked up into a needle of 11 G by use of a syringe and was cooled at room temperature. The charged was extruded to give a cylindrical matrix pharmaceutical preparation (1 mm in diameter, 10 mm in length, about 10 mg in weight).
Example 3 A molten mixture of tetraglycerol dipalmitate and the freeze-dried powdery interferon-a was prepared in the same manner as in Example 1. The molten mixture was sucked up into a 1 ml-syringe (Terumo Co., Ltd., Japan), and, with maintaining the temperature at 50-C, the matrix was sprayed or atomized with an air gun ~Hakuko Co., Ltd., Japan) with extruding from a needle of 27 G ~Terumo Co., Ltd., Japan) to give microspheres.
The microspheres were passed through a sieve ~16 mesh) to remove granular products having a diameter of 1 mm or more.

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~13~86~
, , Example 4 After heated at 48C for melting, the molten tetraglycerol dipalmitate ~300 mg; Sakamoto Yakuhin Kogyo Co., Ltd., Japan) was added with 1.5 mg of a freeze-dried powder of interleukin-2. A portion of the molten mixture was sucked up to a needle of 11 G with a syringe, cooled at room temperature and extruded the charged from the needle to obtain a cylindrical matrix pharmaceutical preparation having a diameter of 1 mm, a length of 20 mm and a weight of about 20 mg.
Example 5 Tetraglycerol dipalmltate ~300 mg; Sakamoto Yaku-hin Kogyo Co., Ltd., Japan) was heated at 48~C for melting, and to the molten was added 1.5 mg of a freeze-dried powder of insulin. The molten mixture was sucked up into a needle of 11 G by use of a syringe and cooIed at room temperature. The charged was extruded from the needle to give a cylindrical matrix pharmaceu-tical preparation ~1 mm in diameter, 20 mm in length, about 20 mg in weight).
Comparative Example 1 A cylindrical matrix pharmaceutical preparation tl mm in diameter, 10 mm in length, about 10 mg in weight) waq obtalned in the same manner as in Example 1 except for using glycerol monopalmitate (the number of ester bond: 1.0; Riken Vitamin Co., Ltd., Japan) instead of tetraglycerol dipalmitate.

- ~1308~

Comparative Example 2 The procedures of Example 1 was followed by using diglycerol monopalmitate (the number of ester bond:
1.0; Sakamoto Yakuhin Kogyo Co., Ltd., Japan) instead of tetraglycerol dipalmitate to give a cylindrical matrix pharmaceutical preparation having a diameter of 1 mm, a length of 10 mm and a weight of about 10 mg.
Comparative Example 3 A cylindrical pharmaceutical preparation (1 mm in diameter, 10 mm in length, about 10 mg in weight) was prepared in the same manner as in Example 2 except for uslng glycerol monostearate ~the number of ester bond:
1.0; Takeda Chemical Industries, Ltd., Japan) instead of tetraglycerol distearate.
Comparative Example 4 The procedure of Example 2 was repeated except for using diglycerol monostearate (the number of ester bond: 1.0; manufactured by Sakamoto Yakuhin Kogyo Co., Ltd., Japan) in place of tetraglycerol distearate to obtain a cylindrical matrix pharmaceutical preparation having a diameter of 1 mm, a length of 10 mm and a ! weight of about 10 mg.
Comparative Example 5 A cylindrical pharmaceutical preparation ~1 mm in diameter, 10 mm in length and about 10 mg in weight) was prepared by the same procedure as in Example 2 except for using tetraglycerol monostearate (the number .~

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of ester bond: 1.0; Sakamoto Yakuhin Kogyo Co., Ltd., Japan) instead of tetraglycerol distearate. -Comparative Example 6 -Using tetraglycerol tristearate (the number of ester bond: 3.0; manufactured by Sakamoto Yakuhin Kogyo Co., Ltd., Japan) instead of tetraglycerol distearate, the procedures of Example 2 was followed to give a cylindrical pharmaceutical preparation (1 mm in diame-ter, 10 mm in length, about 10 mg i~n weight).
Example 6 A cylindrical pharmaceutical preparation (1 mm in dlameter, 10 mm in length, about 10 mg in weight) was prepared in the same manner as in Example 2 except for using tetraglycerol dimyristate (the number of ester bond: 2.0; Sakamoto Yakuhin Kogyo Co., Ltd., Japan) instead of tetraglycerol dipalmitate.
Experimental Example The matrix pharmaceutical preparations obtained in Examples 1 and 2 and Comparative Examples 1 to 6 were respectively administered to a male JCL-SD rat (aged: 6 weeks) subcutaneously in the back in a dose of 10 mg by using a needle of 11 G with a syringe. Each cylindri-cal matrix pharmaceutical preparation contains 9 x 107 International Unit ~IU) of interferon-a.
As a control run, an aqueous solution containing x 107 IU of interferon-a was used.
After administration, 0.6 ml of blood was correct-2130~68 ed from the tail vein with the lapse of time to obtain serum samples. The serum samples were taken respec-tively from three rats, and the concentration of inter-feron-a in each serum sample was determined by sandwich ELISA using two species of anti-interferon-a anti-bodies, and the average value was calculated. As the unit of standard interferon-a, Canferon-TM (Takeda Chemical Industries, Ltd., Japan) was employed. The average values of interferon-a concentration in the serums with the passage of time after administration are set forth in Tables 1 and 2.
In Tables 1 and 2, the term "ND" means "not de-tectable".

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As apparent from Tables 1 and 2, the pharmaceuti-cal preparations of Example 1 and Example 2 respective-ly obtained by using tetraglycerol dipalmitate and tet-raglycerol distearate among polyglycerol higher fatty ~.
5 acid esters can sustainedly release interferon-a for ~-one week or more, and the releasing period of interfer- -on-a is longer than the pharmaceutical preparations of Comparative Examples by a factor of 2 or more.

' ' '

Claims

1. A sustained releasable parenteral pharmaceuti-cal preparation which comprises a matrix containing a physiologically active peptide or protein and a polyg-lycerol diester of a saturated fatty acid.

2. A sustained releasable parenteral pharmaceuti-cal preparation according to claim 1, wherein the polyglycerol has an average polymerization degree of 4.

3. A sustained releasable parenteral pharmaceuti-cal preparation according to claim 1, wherein the saturated fatty acid has 16 to 30 carbon atoms.

4. A sustained releasable parenteral pharmaceuti-cal preparation according to claim 1, wherein the polyglycerol has an average polymerization degree of 4 and the saturated fatty acid has 16 to 30 carbon atoms.

5. A sustained releasable parenteral pharmaceuti-cal preparation according to claim 1, wherein said physiologically active peptide or protein is dispersed in said diester.

6. A sustained releasable parenteral pharmaceuti-cal preparation according to claim 1, wherein the physiologically active peptide or protein has an aver-age molecular weight of 2,000 dalton or more.

7. A sustained releasable parenteral pharmaceuti-cal preparation according to claim 1, wherein the physiologically active peptide or protein is an inter-feron, an interleukin or insulin.

8. A sustained releasable parenteral pharmaceuti-cal preparation according to claim 1, wherein said saturated fatty acid has 16 to 22 carbon atoms.

9. A sustained releasable parenteral pharmaceuti-cal preparation according to claim 1, wherein said saturated fatty acid is palmitic acid or stearic acid.

10. A sustained releasable parenteral pharmaceu-tical preparation according to claim 1, wherein the proportion of the physiologically active peptide or protein is 0.0001 to 50% by weight based on the matrix.

11. A sustained releasable parenteral pharmaceu-tical preparation according to claim 1, wherein said matrix is in a pillar or granular form.

12. A sustained releasable parenteral pharmaceu-tical preparation according to claim 1, wherein said matrix is an injectable solid for implantation.

13. A sustained releasable parenteral pharmaceu-tical preparation which comprises a matrix comprising a physiologically active peptide or protein having an average molecular weight of 5,000 to 1,000,000 dalton and a diester of a polyglycerol having an average poly-merization degree of 4 and a saturated fatty acid having 16 to 22 carbon atoms, wherein the matrix is administrable subcutaneously or intramuscularly and 0.001 to 20% by weight of the physiologically active peptide or protein based on the matrix is dispersed in the diester.

14. A sustained releasable parenteral pharmaceu-tical preparation according to claim 13, wherein said saturated fatty acid is palmitic acid, stearic acid or behenic acid.

15. A sustained releasable parenteral pharmaceu-tical preparation according to claim 13, wherein said diester has a melting point of 40 to 60°C and said matrix is in a solid form at room temperature.

16. A sustained releasable parenteral pharmaceu-tical preparation according to claim 13, wherein said physiologically active peptide or protein is an inter-feron.

17. A method of producing a sustained releasable parenteral pharmaceutical preparation, which comprises mixing a physiologically active peptide or protein with a molten or softened polyglycerol diester of a saturat-ed fatty acid and molding the molten mixture.

18. A method of producing a sustained releasable parenteral pharmaceutical preparation according to claim 17, wherein the molten mixture is molded into a pillar or granular form.

19. A method of producing a sustained releasable parenteral pharmaceutical preparation according to claim 17, wherein the physiologically active peptide or protein in a dried powdery form is mixed with the molten or softened diester.

20. A sustained releasable parenteral pharmaceutical preparation which is solid at room temperature and which comprises a pharmaceutically effective amount of a physiologically active peptide or protein having an average molecular weight of from 5,000 to 1,000,000 dalton and being uniformly dispersed in a matrix composed essentially of a polyglycerol diester of a saturated fatty acid, wherein:
the polyglycerol has an average polymerization degree of 3 to 5;
the polyglycerol diester has an average degree of esterification of from about 1.7 to about 2.3 and a melting or softening point of from about 40 to about 60°C;
the saturated fatty acid has 16 to 22 carbon atoms; and the preparation is produced without using an organic solvent by uniformly mixing the physiologically active peptide or protein with the polyglycerol diester in a molten or softened form and then molding the resulting mixture.

21. The pharmaceutical preparation according to claim 20, wherein the physiologically active peptide or protein is contained in an amount of 0.1 to 10% by weight of the polyglycerol diester.

22. The pharmaceutical preparation according to claim 21, wherein the physiologically active peptide or protein is interferon-.alpha..

23. The pharmaceutical preparation according to claim 22, wherein the polyglycerol diester is tetraglycerol dipalmitate, tetraglycerol distearate or a mixture thereof.

24. The pharmaceutical preparation according to claim 23, which is in the form of an injectable solid for implantation.

25. The pharmaceutical preparation according to claim 23, which is in the form of a suppository.

26. A commercial package which contains therein the pharmaceutical preparation according to any one of claims 1 through 16 or any one of claims 20 through 25, and which carries an indication that the pharmaceutical preparation is a sustained release pharmaceutical preparation.

27. The commercial package according to claim 26, wherein the indication states that an administration thereof may be every seven days or more.

28. A sustained release parenteral pharmaceutical composi-tion which comprises a pharmaceutically effective amount of a physiologically active peptide or protein in a matrix of a polyglycerol diester of a saturated fatty acid.