JPH0372046B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to long-acting interferon formulations. More specifically, the present invention relates to a long-acting preparation that contains interferon as a medicinal ingredient and is designed to maintain blood concentration or intralesional concentration to a clinically useful extent.

Interferon is a type of glycoprotein that is originally produced by animal cells, including humans, when stimulated by viruses or other substances, and is a very useful substance that has virus growth inhibiting and antitumor effects. In recent years, interferon has begun to be used in various clinical applications as a therapeutic agent for viral diseases and malignant tumors, and it is expected that its effects will be enhanced by sustaining its blood concentration or intralesional concentration. However, such a long-acting formulation has not yet been developed.

The present inventors focused on this point and attempted to develop a clinically useful long-lasting interferon preparation, and as a result of intensive studies, the present invention was completed.

That is, it has been extremely difficult to sustain the action of interferon using various conventionally known techniques. This is because interferon is an extremely unstable protein not only in vivo but also as a pharmaceutical product, so any treatment with heat or radiation, or chemical treatment using organic solvents or aldehydes will reduce its activity. This is because interferon preparations have many problems, including the fact that interferon preparations contain only trace amounts of interferon and are water-soluble substances, so they are released extremely quickly after administration into the body. Furthermore, since this preparation is administered parenterally, such as by injection, the accumulation of carriers is a problem, and it is difficult to use carriers that are difficult to decompose in the body, such as silicone, which has recently been used in some medical applications. There were even more restrictions. Under such circumstances,
As a result of extensive research over many years, the present inventors have overcome many of the difficulties and problems described above by using the method of the present invention, and have obtained a long-lasting preparation that is expected to be extremely useful clinically. This completes the present invention.

That is, the present invention is a long-lasting preparation of interferon, which is characterized by containing interferon in a low-toxic carrier - collagen, which is biodegradable and implantable in the body. Blood or local concentrations are maintained to a useful extent.

Now, regarding interferon, which is a medicinal ingredient, it may be used alone, together with an interferon activator, or with other medicinal ingredients that can be expected to have an additive or synergistic effect with interferon. It may be a combination of Note that the interferon referred to herein may be any interferon such as α, β, γ, or a combination thereof.

As for the carrier, collagen or a mixture of collagen and gelatin is preferably used from the viewpoint of safety and ease of use. Collagen is the main protein in animal connective tissue.
It is a safe protein that has low antigenicity and is already frequently used in medical surgical threads.

Furthermore, for the purpose of further increasing safety, atelocollagen with lower antigenicity may be used by removing the telopeptide portion by treating the collagen with an enzyme, for example, with pepsin. Gelatin is also a protein derived from collagen.
It has already been evaluated for medical safety as an inexpensive polyampholyte with low antigenicity and sol-gel conversion properties.

Next, an example of a method for incorporating interferon into the carrier of the present invention will be explained. That is, the interferon-containing solution and the carrier-containing solution are mixed and stirred as uniformly as possible without forming bubbles, and if necessary, concentrated at a low temperature, or optionally spray-dried or freeze-dried. At this time, pharmaceutically acceptable stabilizers, preservatives, soothing agents, etc., and additives for adjusting moldability and sustained release properties can be added as necessary. The material thus obtained is processed as appropriate depending on the purpose. For example, it is crushed under cooling with dry ice, liquid nitrogen, etc. to obtain a powder. At this time, the particles crushed and micronized to an injectable particle size can be suspended in a viscous injection solvent to form a long-lasting suspension injection. Alternatively, by separately adding a viscous solvent for injection, a preparation can be prepared that is suspended before use. The interferon and the carrier may be atomized by other general methods as appropriate. Viscous injection solvents include - for example, sesame oil, peanut oil, cottonseed oil, MCT (medium chain fatty acid triglycerides), olive oil, corn oil, castor oil, silicone oil, PEG (polyethylene glycol), PG (propylene glycol), Iodinated poppy oil fatty acid ethyl ester, etc.
It refers to In addition, fine particles pulverized to an appropriate particle size by the above-mentioned method or other appropriate method are collected, and if necessary, additives for molding are added and compression molded, which can be administered through a fiberscope forceps needle or an indwelling needle. Shape or rod shape (diameter 0.5mm
~1.5 mm, length approximately 5 mm to 15 mm).
Alternatively, it can be placed in a mold in advance, concentrated or freeze-dried at low temperature, and similarly compressed to form a needle- or rod-shaped preparation. Furthermore,
A similar method can be used to prepare an implantable preparation, that is, a preparation in the form of pellets, spheres, granules, powder, etc., which can be implanted or dispersed into the body or lesion during surgery. These can be used depending on the situation, such as the degree of duration needed at that time, but it is natural that the larger the shape, the longer the duration.

It goes without saying that each of these steps is performed aseptically because of the nature of the injection or implant.

Next, the present invention will be explained more clearly by experimental examples and examples, but these examples are not intended to limit the present invention.

Experimental Example 1 The collagen oil-based suspension preparation prepared in Example 1 (sample a), the collagen acicular preparation prepared in example 2 (sample b), and α-
Aqueous injections (sample c) of interferon (derived from Namalva cells) were each administered intramuscularly to rabbits, and the time course of the blood concentration was examined using quantitative determination using the R method. Two rabbits were used each,
The dose was 10 ⁶ U/Kg. The average value of two birds was used for the numerical value.

The results are shown in Figure 1. As can be seen in Figure 1, samples a and b showed a tendency to persist, with blood concentrations of several + U/ml being maintained even after 48 hours, with the needle-like preparations in particular being more persistent.

In this way, in vitro experiments using domestic rabbits also suggested the clinical usefulness of the formulation of the present invention.

Example 1 Solution containing α-type interferon (potency
4.9MU/ml) 100ml and 2% atelocollagen 50
The mixture is mixed and stirred uniformly with as little foam as possible, freeze-dried, and then cryogenically ground using liquid N ₂ . By suspending this in sesame oil
An oil-based suspension type long-acting preparation containing 4 MU of interferon per vial was obtained. (Sample a) Example
2 By compression molding the pulverized product obtained in Example 1, a needle-shaped long-acting preparation containing 10 MU of interferon per bottle was obtained. (Sample b) Example 3 The pulverized product obtained in Example 1 was suspended in castor oil to obtain an oil-based suspension type long-acting preparation containing 4 MU of interferon per vial.

Example 4 The pulverized product obtained in Example 1 was compression-molded into pellets to obtain a pellet-like long-acting preparation containing 10 MU of interferon per pellet.

Example 5 Solution containing α-type interferon (potency
4.9MU/ml) 100ml and 2% atelocollagen 50
g, human serum albumin 150mg and thimerosal
120 μg was mixed and stirred uniformly with as little foam as possible, freeze-dried, and then cryogenically ground using liquid N ₂ . By suspending this in sesame oil, an oil-based suspension type long-acting preparation containing 4 MU of interferon per vial was obtained.

Example 6 Solution containing α-type interferon (potency
Mix 100ml of 4.9MU/ml) and 50g of 2% collagen as homogeneously as possible without forming bubbles.
After freeze-drying, cryogenically grind using liquid _N2 . By suspending this in sesame oil, per 1 vial,
An oil-based suspension type long-acting preparation containing 4MU of interferon was obtained.

Example 7 Solution containing α-type interferon (potency
4.9MU/ml) 100ml and atelocollagen powder 1g
After mixing and dissolving by adding 0.1N hydrochloric acid,
Pour into molds and freeze dry. By compression molding this, a needle-like long-acting preparation containing 10 MU of interferon was obtained.

Example 9 The pulverized product obtained in Example 1 was suspended in PEG to obtain a suspended long-acting preparation containing 4 MU of interferon per vial.

Example 9 To the pulverized product obtained in Example 1, 25% by weight of methylcellulose was added, and distilled water for injection was added to give a solid content of 20%, followed by kneading. This is put into a mold, freeze-dried, and then compression molded.
A needle-like long-acting preparation containing 10 MU of interferon was obtained.

Example 10 Solution containing α-type interferon (potency
4.9MU/ml) 100ml and 2% atelocollagen 50g
Mix and stir 98 mg of tespamine evenly, without forming bubbles as much as possible, and freeze-dry.
Cryogenically grind using _N2 . By compression molding this, a needle-like long-acting preparation containing 10 MU of interferon and about 2 mg of tesbamine was obtained.

Example 11 By suspending the pulverized product obtained in Example 1 in iodinated poppy oil fatty acid ethyl ester (Lipiodol Ultrafluid, handled by Kodama Shoji), an oil-based suspension containing 4 MU of interferon per vial was prepared. Long-acting formulations can be obtained.

[Brief explanation of drawings]

Figure 1 shows the changes in blood concentration after intramuscular administration to domestic rabbits, showing the two formulations of the present invention and α-
This is a comparison with interferon aqueous injection. Vertical axis: Blood concentration of α-interferon Unit: unit/ml Horizontal axis: Time Unit: time ○; Collagen oil suspension of the present invention ▲; Collagen needle-like preparation of the present invention ●; α-Interferon aqueous injection agent

Claims (1)

[Scope of Claims] 1. A long-lasting interferon preparation characterized by containing interferon in a carrier made of collagen. 2. The long-acting preparation according to claim 1, wherein the interferon and the carrier are atomized and suspended in a viscous injection solvent. 3. The long-acting preparation according to claim 1, wherein the interferon and the carrier are formed into a rod or needle shape. 4. The long-acting preparation according to claim 1, wherein the interferon and the carrier are formed into granules, powders, spheres, pellets, or the like.