JPH0725688B2 - CSF sustained release formulation - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a sustained-release preparation. More specifically, it relates to a sustained-release preparation characterized by containing colony stimulating factor (CSF) as an active ingredient and being devised so that its blood concentration or intralesional concentration is maintained for a clinically useful period for a long period of time. is there.

[Conventional technology and its problems]

CSF is a general term for physiologically active substances that stimulate proliferation and differentiation of hematopoietic stem cells in bone marrow and promote colony formation of macrophage polynuclear leukocytes in vitro, but it is becoming clear that CSF also acts as a hematopoietic factor in vivo. With the progress of research on such physiological effects, clinical utility of each subclass of CSF has been expected more specifically. For example, chemotherapy for leukemia and cancer and bone marrow transplantation therapy cause granulocytopenia due to bone marrow hypoplasia, but it is expected that administration of CSF will accelerate recovery. Since these symptoms last for several days or longer, it is expected that the clinical effect will be enhanced by the sustained action of CSF. However, such a sustained-release CSF preparation has not been developed yet.

[Structure of Invention]

The present inventors have completed the present invention as a result of diligently studying and attempting to develop a clinically useful CSF preparation with attention paid to this point. That is, a sustained-release solid preparation characterized by sustained release over a long period (for example, several weeks to several months) by containing an active ingredient in a carrier made of collagen or gelatin or a mixture of collagen and gelatin is completed. It was done.

CSF which is an active ingredient may be CSF alone, may be present together with an activity enhancer of CSF, or may be a combination of substances expected to have an additive or synergistic effect with CSF.

The CSF here is recognized as a CSF family and is defined as a physiologically active substance that stimulates proliferation and differentiation of hematopoietic stem cells of bone marrow in vitro and promotes colony formation of macrophage polynuclear leukocytes. . CSF
There are various subclasses of erythrocyte, and pluripotent CSF (multi-CSF), granular-monocyte macrophage CSF (GM-CSF), and granulocyte CSF (G-CS) are present depending on the type of colony formed.
F) and monocyte macrophage CSF (M-CSF). It is also considered that even in the same CSF subclass, the charge and the molecular weight may differ mainly because the sugar chains are different. Further, although there is a report that multi-CSF is the same substance as interleukin-3 (IL-3), any substance may be used in the present invention and a mixture thereof may be used. In any case, it is expected that each subclass whose structure is expected to be determined by future studies is a glycoprotein or protein having a similar molecular weight, and it is considered that any sustained release can be achieved by applying the present invention. . The CSF used in the present invention may be a substance extracted from a living body, an artificial synthetic substance, or a substance obtained by a gene recombination method regardless of its production method.

From the viewpoint of safety and ease of use, it is preferable to use collagen or gelatin, or a mixture of both. Collagen is a major protein of connective tissue of animals, and is a safe substance which is already used medically as a protein with little antigenicity such as surgical thread.

For the purpose of further improving safety, atelocollagen whose antigenicity is further reduced by removing the telopeptide portion by enzymatic treatment of collagen, for example, treatment with pepsin may be used. Gelatin is an inducible protein from collagen and is an inexpensive polyampholyte with low antigenicity and sol-gel conversion property, and its medical safety evaluation has already been established. Can be used.

In the present invention, the active ingredient and carrier are preferably pure products, but those which are usually available may be used. Those which are usually available usually contain suitable stabilizers, buffers and other additives in suitable amounts. For example, the collagen aqueous solution usually contains a buffer solution of an inorganic salt or an organic salt such as a phosphate buffer solution, a citrate buffer solution, an acetate buffer solution, or the like. Commonly available CSF includes, for example, buffer, sodium chloride, human serum albumin, amino acids (eg glycine, alanine etc.), sugars (eg glucose etc.), sugar alcohols (eg mannitol, xylitol etc.), polyethylene glycol etc. Has been. These may be used as they are, but it is desirable to remove such additives and other components from the viewpoint of sustained release.

The mixing ratio of the active ingredient and the carrier is not particularly limited, but for example, CSF 10 ⁴ to 10 ⁹ U is preferable per 1 mg of the carrier. CSF
As a fitting, 10 ⁵ to 10 ¹⁰ U per formulation, usually 10 ⁶ to 10 ⁹
It should be U.

The sustained-release preparation of the present invention can be obtained, for example, by the following method.

The active ingredient or its aqueous solution and the carrier or its aqueous solution are stirred and mixed. That is, the active ingredient is included in the carrier matrix by mixing the active ingredient and the carrier in a solution state. The mixture is then concentrated and / or dried. Although the concentration method and the drying method are not particularly limited, examples of the concentration method include a method of standing at room temperature and the like, and examples of the drying method include natural drying, spray drying and freeze drying. The above steps are usually carried out at room temperature or lower. It may be cooled down. As a concrete aim, for example, the mixing is usually about 5 to 30 ° C., and the lyophilization is usually about -50 to 0.
Drying by air-drying or spray drying is usually carried out at room temperature or lower (for example, about 15 to 30 ° C). In the case of spray drying, the temperature of the active ingredient can be controlled to 40 ° C or less by keeping the solution temperature and the sample container temperature below room temperature and controlling the drying conditions, and it is possible to prevent the active ingredient from being substantially damaged. it can.

The method described above is suitable for heat-labile CSF because it requires no special binder and no heat is applied.

The sustained-release preparation of the present invention preferably consists essentially of the active ingredient and a biodegradable carrier. Since the presence of components other than the active ingredient and the carrier often promotes the release of the active ingredient, it is preferable to exclude such other ingredients as much as possible. However, in reality, even if an active ingredient or other ingredient derived from a carrier that is usually available is contained, it may be contained within a range that does not substantially affect the sustained-release effect. Similarly, the sustained-release preparation of the present invention contains, if necessary, conventional pharmaceutically acceptable additives such as stabilizers, preservatives and local anesthetics, etc. within a range that does not substantially affect the sustained-release effect. May be included.

The thus obtained material is appropriately processed according to the purpose. For example, with dry ice or liquid nitrogen, the formulation is about -10 ~
While cooling so as to be kept at -100 ° C, or at room temperature or lower, pulverize by a usual method into powder. It is compression molded by a mold. Alternatively, the mixture of the active ingredient and the carrier may be put in a mold in advance, and then concentrated or air-dried or freeze-dried, and finally compression-molded. Also, injection molding can be performed by a usual method.

At this time, as the shape of the preparation, for example, spherical, hemispherical, columnar, needle-like, rod-like, button-like, etc. are molded into a shape suitable for the site to be used, and subcutaneous administration, implantation in a living body or insertion into a body cavity is performed. It can be administered. As a guide for the size, for example, in the case of a needle or a rod, the diameter is 0.5 to 5 mm and the length is 5 to 50.
mm, preferably 0.5 to 1.5 mm in diameter and 5 to 15 mm in length.

Next, the present invention will be described more clearly by way of examples and experimental examples, but none of these examples limit the present invention.

Example 1 0.2 ml of an aqueous solution of mouse GM-CSF (containing 2 × 10 ⁵ U, including human serum albumin and sodium chloride) was mixed with 2 ml of a 2% atelocollagen phosphate buffer solution, followed by freeze-drying. . This was pulverized at a low temperature using liquid nitrogen, put into a mold and compression-molded to obtain a cylindrical sustained-release preparation.

Example 2 To 500 mg of collagen powder, 3 ml of a mouse GM-CSF aqueous solution (containing 5 × 10 ⁵ U, human serum albumin and sodium chloride were also added) was swollen. The obtained gel was defoamed, extruded, naturally dried and cut to obtain pellets of a sustained-release preparation (diameter: 1.2 mm, length: 10 mm).

Experimental Example 1 The pellet obtained in Example 2 was placed in a physiological saline solution, and GM-CSF released from the preparation was quantified by HPLC to determine the cumulative release rate.

The results are shown in Fig. 1.

[Brief description of drawings]

FIG. 1 shows the cumulative release rate of the sustained-release preparation of mouse GM-CSF of Experimental Example 1. The vertical axis represents the cumulative release rate (%) and the horizontal axis represents the elution time (hour).

Continuation of the front page (56) Reference JP-A-60-112713 (JP, A) JP-A-59-130252 (JP, A) JP-A-57-150609 (JP, A) JP-A-61-277612 (JP , A)

Claims (5)

[Claims] 1. A sustained-release solid preparation characterized in that a colony-stimulating factor is contained in a carrier made of collagen or gelatin or a mixture of collagen and gelatin. 2. A colony stimulating factor (CSF) is a pluripotent CSF (mu).
lti-CSF), granulocyte-monocyte macrophage CSF (GM-CS)
F), granulocyte CSF (G-CSF) or monocyte macrophage C
The sustained-release solid preparation according to claim 1, which is SF (M-CSF). 3. The sustained-release solid preparation according to claim 1, wherein the colony stimulating factor (CSF) is granulocyte-monocyte macrophage CSF (GM-CSF). 4. The colony stimulating factor (CSF) is granulocyte CSF (G
-CSF) The sustained-release solid preparation according to claim 1. 5. A carrier containing an active ingredient is spherical,
The sustained-release solid preparation according to claim 1, 2, 3, or 4, which is molded into a hemispherical shape, a cylindrical shape, a needle shape, a rod shape, or a button shape.