JPS6227048B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention provides a method for transplanting interferon, particularly cells of human origin in culture, into a warm-blooded non-human animal, or in a diffusion chamber attached to or outside a warm-blooded non-human animal. The human-derived cells are grown while being supplied with the body fluids of the warm-blooded animal, and an interferon inducer is applied in vivo or in vitro to produce interferon, and the purified and fractionated interferon is used as the active ingredient. The present invention relates to an anti-interferon-sensitive disease agent, that is, an agent for preventing and/or treating interferon-sensitive diseases. Interferon is "Interferon" by Shigeyasu Kobayashi, 1975, published by Kodansha Co., Ltd., DAJ
“INTERFERON and its Clinical
“Potential” 1976 William Heinemann Medical
Published by Books Ltd. (London), “Protein Nucleic Acid Enzyme
Vol. 21, No. 4'' (1976), for example, when interferon inducers such as viruses, bacteria, protozoa, rickettsiae, nucleic acids, endotoxins, and polysaccharides act on living cells. Therefore, it is a name given to a protein-like substance that is induced and produced inside and outside the cell and has the function of non-specifically suppressing the proliferation of various viruses within the cell. Because of these functions of interferon,
Since its discovery, interferon has been expected to be a preventive and therapeutic agent for viral diseases. Furthermore, in recent years, interferon has been shown to have antitumor properties not only against viral tumors but also against non-viral tumors, and interferon has come to be widely used as a drug. However, interferon is highly species-specific, and in order to prevent or treat human diseases, it is necessary to
It is not effective unless interferon is produced from living human cells. Live human cells traditionally used for the preparation of interferon include white blood cells. However, leukocytes are prepared by separating them from fresh human blood, and it is difficult to preserve them, and it is extremely difficult to supply them in large quantities at low cost. In addition, attempts have also been made to use cultured human-derived cells inoculated into a nutrient medium in vitro to increase their color. However, the method of growing in vitro not only requires a large amount of expensive human serum in the nutrient medium, but also its growth is unstable and its growth rate is low.
Furthermore, there are drawbacks such as the low concentration of cells obtained, making it extremely difficult to stably supply large quantities of living human cells at low cost. Due to the current situation as described above, the production of interferon that can be used for the prevention and treatment of human diseases has not yet been carried out on an industrial scale. The present inventors have developed a method for producing interferon that can be easily carried out on an industrial scale, and the interferon is useful as an anti-interferon-sensitive disease agent, that is, as a prophylactic and therapeutic agent for interferon-sensitive diseases. I did a lot of research on whether or not. As a result, the cultured human-derived cells,
Rather than being inoculated and grown in a nutrient medium outside the body, it is transplanted into the body of a warm-blooded non-human animal, or in a diffusion chamber installed outside or within the body of a warm-blooded non-human animal. Interferon, which is highly species-specific, is highly active in humans by multiplying the resulting human-derived cells while receiving a supply of body fluid containing It was discovered that interferon can be easily produced in large quantities by purifying and fractionating the induced product, and that interferon can be used as an anti-interferon-sensitive disease agent, that is, as a prophylactic or therapeutic agent for interferon-sensitive diseases. After confirming this, the present invention was completed. Unlike when living cells are grown in vitro, the method for producing interferon used in the present invention not only eliminates or significantly saves a nutrient medium containing expensive blood semen, but also eliminates the need for a nutrient medium containing expensive blood sperm. It is extremely easy to maintain and manage, and it also has the characteristics of high induced interferon activity. In other words, if cultured human-derived cells are transplanted into the body of a warm-blooded animal other than humans, or housed in a diffusion chamber that can receive body fluids from that animal, and reared normally, they will become warm-blooded. The cells can easily proliferate using body fluids containing nutrients provided by the animal body. Furthermore, compared to when grown in vitro, the proliferation of these cells is stable and the proliferation rate is high;
It is characterized by a large amount of cells obtained and further by a high yield of interferon per cell. The cultured human-derived cells used in the method for producing interferon used in the present invention may be any cell that can be transplanted into the body of a warm-blooded animal other than humans and easily proliferated. "Protein Nucleic Acid Enzyme Vol. 20 No. 6" pp. 616-643 (1975)
OUMS-20 cells, OUMS-25 reported in
Even if it is an established cell line derived from normal human cells such as HEF cells, HUF-2 cells, WI-38 cells, etc.,
Vol. 1” pp. 116-117 (1975)
Namalva cells, “Nature Vol.267” by I. Miyoshi
BALL- reported on pages 843-844 (1977)
1 cell, TALL-1 cell, NALL-1 cell, I.
Written by Miyoshi “Cancer Vol.40 No.6” No. 2999~
Even cell lines derived from human tumor cells such as JBL cells reported on page 3003 (1977) may be freely used, and the invention is not limited to the cell lines described herein. The warm-blooded animal used in the method for producing interferon used in the present invention may be any animal that can proliferate human-derived cells, such as birds such as chickens and pigeons, dogs, cats, monkeys, goats, pigs, etc. Mammals such as cows, horses, rabbits, guinea pigs, rats, hamsters, regular mice, and nude mice can be used. Transplanting human-derived cells into these animals may cause an unfavorable immune reaction, so in order to suppress the reaction as much as possible, the animals used should be kept as young as possible, i.e. eggs, embryos, fetuses, etc.
Or those in the newborn stage or early childhood are preferable. In addition, these animals may be irradiated with X-rays or gamma rays of approximately 200 to 600 rem, or
Alternatively, transplantation may be performed after pretreatment such as injection of antiserum or immunosuppressant to weaken the immune response. When the animals used are nude mice, the immune response is weak even when they are grown, so cultured human-derived cells can be transplanted without the need for these pretreatments, and the cells can be rapidly grown. This is particularly advantageous since it can be propagated. In addition, human-derived cells that have been cultured are first transplanted into hamsters and allowed to proliferate, and then these cells are further transplanted into nude mice.
It is also possible to stabilize the growth of human-derived cells by transplanting them between warm-blooded animals other than humans, or to increase the amount of interferon induced therefrom. In this case, transplantation may be performed not only between the same species and the same genus, but also between the same order and phylum. The site in the animal body to which human-derived cells are transplanted may be any site where the transplanted cells can proliferate, such as the allantoic cavity, veins,
The abdominal cavity, subcutaneous, etc. can be freely selected. In addition, porous filtration membranes that can block the passage of animal cells without directly transplanting human-derived cells into the animal body, such as membrane filters with pore diameters of about 10 ^-7 to ^{10 -5} m, ultrafiltration Diffusion chambers of various known shapes and sizes provided with membranes or hollow eye bars are buried in the animal body, for example, in the abdominal cavity, and the diffusion chamber is supplied with body fluids containing nutrients from the animal body. Any of the human-derived cells cultivated as described above can be grown. In addition, if necessary, a diffusion chamber that connects and perfuses the solution containing nutrients in the diffusion chamber with body fluids within the animal body may be attached to the surface of the animal body, and human-derived cells within the diffusion chamber may be perfused. It is possible to see through the growth state, and it is also possible to make only the diffusion chamber part detachable and replaceable, allowing cells to proliferate to the fullest lifespan without killing the animal, thereby further increasing the cell production amount per individual animal. You can also do it. These diffusion chamber-based methods do not allow direct contact between human-derived cells and animal cells;
Not only can only human-derived cells be easily collected, but there is also little risk of causing undesirable immune reactions, so there is no need for pretreatment to suppress immune reactions.
It has the characteristic of being able to freely utilize various warm-blooded animals. The maintenance and management of transplanted animals is convenient because it is sufficient to continue the normal breeding of the animal, and no special handling is required even after transplantation. The period for growing human-derived cells is usually 1 to 20 weeks. When the cultured cells to be transplanted are derived from human normal or neoplastic leukocytes, their proliferation rate is particularly high, usually 1.
The goal can be achieved in a period of ~5 weeks. It has also been found that the number of human-derived cells obtained in this manner reaches approximately 10 ⁷ to 10 ¹² or more per animal. In other words, the number of human-derived cells proliferated by the interferon production method used in the present invention is approximately 10 ⁷ to 10 ¹² of the number of cells transplanted per individual animal.
It can reach about 10 ¹ to 10 ⁶ times or more than inoculation and propagation in in vitro nutrient medium, which is very convenient for the production of interferon. Any method can be used to induce and produce interferon from human-derived living cells grown in this manner. The interferon-inducing agent can also be applied to the interferon in the animal body in which it has grown.
For example, human-derived cells grown in suspension in ascites fluid in the peritoneal cavity, or tumor cells generated subcutaneously,
Interferon may be induced and produced by direct action of an interferon-inducing agent, and then interferon may be purified and isolated from the ascites or tumor. Furthermore, human-derived proliferating cells can be removed from an animal body and interferon can be induced and produced by treating them with an interferon-inducing agent outside the body. For example, human-derived cells grown in ascites are sorted, or subcutaneous tumors containing human-derived cells are excised and dispersed, and the resulting cells are approximately 20 to
The cell concentration in the nutrient medium kept at 40°C is approximately 10 ⁵ - 10 ⁸ /
ml, and then treated with an interferon-inducing agent to induce and produce interferon, which can then be purified and fractionated. Furthermore, when human-derived cells are grown in a diffusion chamber, the grown cells can be left in the diffusion chamber or removed from the diffusion chamber and treated with an interferon-inducing agent to induce interferon production. You can also do it. In addition, during the induced production of interferon,
For example, the amount of interferon produced can be further increased by priming using interferon, which is highly specific to humans, or by employing known methods such as superinduction using a metabolic inhibitor. Be free. In addition, for example, interferon can be induced and produced in human-derived cells that have been proliferated in the animal body, and then interferon can be produced outside the animal body in human-derived cells collected from a specific part or whole of the same animal. A method of inductively producing interferon, a method of using cells once used for inductively producing interferon two or more times, or cells obtained by replacing the diffusion chamber buried or connected in the animal body. It is also possible to further increase the amount of interferon produced per individual animal used by methods such as increasing the number of animals used. Known interferon inducers such as viruses, bacteria, protozoa, rickettsiae, nucleic acids, endotoxins, polysaccharides, etc. can be freely used. Interferon thus induced and produced can be purified by known purification and separation methods such as salting out, dialysis,
It can be easily purified and separated and collected by filtration, centrifugation, concentration, freeze-drying, etc. If a higher level of purification is required, known methods such as adsorption/elution on an ion exchanger, gel filtration, affinity chromatography, high performance liquid chromatography, isoelectric focusing, and electrophoresis can be used. It is also possible to collect interferon of the highest purity by further combining them. The activity of interferon, which is highly specific to humans, is described in "Protein Nucleic Acid Enzyme Vol. 20 No. 6" p. 616 -
643 (1975), derived from human amnion.
It was measured by a known plaque half-life method using FL cells. The hemagglutination titer is determined by JESalk, “The Journal of
Jmmunology Vol. 49, pp. 87-98 (1944). Next, a few methods for producing interferon used in the present invention will be described. Production Example of Interferon 1 After subcutaneously transplanting cultured human-derived BALL-1 cells into adult nude mice, they were raised in the usual manner for 3 weeks. Approximately 10 g of tumor that had occurred under the skin was excised and cut into small pieces, and then suspended in trypsin-containing physiological saline to disperse and separate the cells. The cells were washed with Eagle's minimal basic medium of pH 7.2 containing 5v/v% human serum, and diluted to a cell concentration of approximately 5 x 10 ⁶ /ml in a medium of the same composition kept at 37°C.
To this, partially purified human interferon, which is highly species-specific, is added at a rate of approximately 100 units/ml, resulting in approximately 2
It kept time. Sendai virus was added to this at a rate of about 300 hemagglutination titer/ml and maintained for 20 hours to induce interferon production. This is about 4℃, about
Centrifugation was performed at 1000 g to remove precipitates such as cells, and the resulting supernatant was dialyzed against pH 2.0, 0.1 M potassium hydrochloride buffer at 4°C for 48 hours, then pH 7.2,
12 in saline containing 0.01M phosphate buffer
The filtrate obtained by time dialysis and further microfiltration was concentrated and freeze-dried to obtain a powder having interferon activity. The obtained interferon activity was
It was approximately 20,000,000 units per nude mouse. Production Example of Interferon 2 Human-derived OUMS-20 cells cultured in the peritoneal cavity of adult nude mice were transplanted and then raised in a conventional manner for 5 weeks. New Katsuru disease virus with a hemagglutinating value of about 3000 was injected into the abdominal cavity after being inactivated by ultraviolet rays, and after 24 hours, the virus was killed and ascites was collected. This is 4℃, about 1000
g to remove precipitates such as cells, and the resulting supernatant was dialyzed against PH2.0, 0.1M potassium hydrochloride buffer at about 4°C for 48 hours.
7.2, dialyzed against physiological saline containing 0.01M phosphate buffer for 15 hours, and further microfiltered, and the resulting filtrate was concentrated to obtain a solution having interferon activity. The interferon activity obtained was approximately 700,000 units per 10 nude mice. Production example of interferon 3 A newborn hamster was injected with an antiserum prepared from a rabbit using a known method to weaken the hamster's immune response, and then cultured human-derived JBL cells were subcutaneously transplanted into the hamster. Thereafter, the animals were raised in the usual manner for 4 weeks. After about 30 g of tumor that had occurred under the skin was removed, the cells were dispersed in the same manner as in Production Example 1. These cells were added to PH7.4 containing 10% v/v of calf serum.
After washing with RPMI1640 medium, the cells were diluted in a medium of the same composition kept at 37°C to a cell concentration of approximately 2×10 ⁷ /ml. To this, partially purified human interferon, which is highly species-specific, was added at a rate of 200 units/ml and kept for about 1 hour, and then Sendai virus was added at a rate of about 100 hemagglutination titer/ml.
After a period of time, interferon was induced to be produced. Thereafter, the solution was purified and concentrated in the same manner as in Production Example 2 to obtain a solution having interferon activity. The interferon activity obtained was approximately
It was 1,500,000 units. Production Example of Interferon 4 Neonatal rats were intravenously transplanted with cultured human-derived Namalva cells and then reared for 4 weeks in a conventional manner. After about 50 g of the tumor that had formed under the skin was removed, the cells were dispersed in the same manner as in Production Example 1. Next, it was treated in the same manner as in Production Example 1 to induce the production of interferon, further purified in the same manner as in Production Example 1, and freeze-dried to obtain a powder having interferon activity. The activity obtained was approximately
It was 30000000 units. Production example of interferon 5 After pre-irradiating an adult normal mouse with X-rays of about 400 rem to weaken the mouse's immune response, a human-derived human-derived strain cultured under the mouse's skin is
Transplant TALL-1 cells, and then use standard methods for 3
They were kept for a week. After about 10 g of the tumor that had formed under the skin was removed, the cells were dispersed in the same manner as in Production Example 1. These cells were treated in the same manner as in Production Example 3 to induce the production of interferon, and thereafter purified and concentrated in the same manner as in Production Example 2 to obtain a concentrated solution having interferon activity. The obtained interferon activity was
It was approximately 8,000,000 units per mouse. Interferon Production Example 6 Cultured OUMS-25 cells were first transplanted subcutaneously into hamsters using the method described in Production Example 3, and the cells obtained by growing for 3 weeks were then transplanted into 10-day-old nude mice. It was implanted intraperitoneally. After the nude mice were raised in a conventional manner for 5 weeks, they were anesthetized and ascites fluid was collected and centrifuged to obtain proliferating cells. After washing the cells by the method of Production Example 1, they are treated in the same manner as in Production Example 1 to induce and produce interferon, and then purified and concentrated in the same manner as in Production Example 2 to obtain a concentrated solution having interferon activity. Ta. The interferon activity obtained was approximately
It was 2,000,000 units. Interferon production example 7 Human-derived JBL cultured in a plastic cylindrical diffusion chamber with an internal capacity of about 10 ml and equipped with a membrane filter with a pore size of about 0.5 microns.
The cells were suspended in physiological saline and implanted into the peritoneal cavity of an adult rat. After raising these rats in the usual manner for 4 weeks,
The diffusion chamber was removed. The concentration of human-derived cells obtained in this way was about 5 x 10 ⁹ /ml, which is about the same as when grown in an in vitro nutrient medium in a carbon dioxide incubator.
It was found that the increase was more than 10 ³ times. The cells were treated in the same manner as in Production Example 1 to induce the production of interferon, purified, concentrated, and lyophilized to obtain a powder having interferon activity.
The interferon activity obtained was approximately 30,000,000 units per rat. Interferon Production Example 8 Cultured human-derived NALL-1 cells were transplanted into fertilized chicken eggs kept at 37°C for 5 days, and then kept at 37°C for 1 week. After breaking the eggs, the proliferating cells were collected, and the cells were treated in the same manner as in Production Example 1 to induce and produce interferon, and then purified and concentrated in the same manner as in Production Example 2 to detect interferon activity. A concentrated solution having the following properties was obtained. The interferon activity obtained was approximately 400,000 units per 10 fertilized eggs. Production example of interferon 9 Interferon powder prepared by the method of production example 1 was prepared in the report of G. Bodo (Symposium on
Preparation, Standardization and Clinical Use
of Interferon.11th International
Immunobiological Symposium. 8&9
June 1977 ^, Zagreb. Purity interferon was obtained with a yield of about 40%. The interferon used in the present invention obtained as in the above-mentioned production example may be interferon alone or interferon combined with one or two kinds of interferon.
The following therapeutic experiments revealed that by incorporating other substances of more than 1 species, it can be used advantageously for the prevention and treatment of interferon-sensitive diseases in humans, for example, as injections, external medicines, and oral medicines. . Experiment 1 Treatment of viral diseases with interferon (in vitro virus growth inhibition test) Interferon production example 9 was applied to human fetal lung primary culture cells cultured in a monolayer in a 6 cm diameter Shear
Interferon 1, 10 or
After adding 100 units and keeping it in a 5% carbon dioxide gas incubator at 37°C for 20 hours, a quantity of Valicellazoster virus (which has the ability to generate approximately 100 plaques without interferon) was added. The number of plaques generated by adding varicella-zoster virus) or human cytomegalovirus (a virus that causes stillbirth and premature birth) was counted. The virus proliferation inhibiting effect was determined based on the rate of reduction in plaque number caused by interferon. Plaque number reduction rate (%) = AB/A x 100 A: Number of plaques without the addition of interferon B: Number of plaques with the addition of interferon The counting results are shown in Table 1 below.

[Table] As is clear from the results in Table 1, it can be seen that the interferon used in the present invention effectively inhibits the proliferation of viruses that cause viral diseases. In addition, no abnormality was observed in human cultured cells due to the addition of interferon. Experiment 2 Treatment of non-viral diseases with interferon (1) In vitro tumor cell growth inhibition test Interferon prepared by the method of Interferon Production Example 9 was added to RPMI1640 medium containing 15 v/v% fetal bovine serum. The final concentration is 30, 300, 3000
units/ml, and inoculated with various tumor cells at a concentration of 5 x 10 ⁵ /ml. After culturing for 5 days in a 5% carbon dioxide gas incubator kept at 37°C. , the number of cells per ml of medium was determined. As a control, interferon that had been heat-inactivated by keeping it at 100°C for 30 minutes was added in equal amounts, cultured in the same way, and measured. The cell proliferation inhibition rate was determined using the following formula. Cell proliferation inhibition rate (%) =
(A-5×10 ⁵ )-(B-5×10 ⁵ )/(A-5×
10 ⁵ )×100 A: Number of cells in the control group B: Number of cells in the test group The results of the measurements are shown in Table 2 below.

[Table] As is clear from the results in Table 2, interferon BALL-1 cells used in the present invention,
It can be seen that the proliferation of tumor cells such as TALL-1 cells, NALL-1 cells, and JBL cells is significantly inhibited, and that the active concentration is effective in the range of 30 to 3000 units/ml. (2) In vivo tumor cell growth inhibition test The test was conducted using 8 nude mice approximately 2 months old. 7.5×10 ⁶ tumor cells TALL-1 per animal were subcutaneously implanted in all eight animals. Interferon production example 3 was applied to 4 of these animals from 2 days after transplantation.
Interferon solution prepared by the method of
10,000 units were injected intraperitoneally three times a week for a total of 20 times, and on the 48th day after transplantation, the fresh weight of the resulting tumor mass was measured. The remaining 4 mice were raised in the same manner as the previous 4 mice, except that interferon was not injected as a control, and 48 days after transplantation, they were sacrificed and the fresh weight of the resulting tumor mass was measured. The measured results are shown in Table 3 below.

[Table] (3) In vivo tumor cell proliferation inhibitory effect test The test was conducted using 8 nude mice approximately 2 months old. Approximately 10 ⁷ tumor cells JBL per animal were subcutaneously implanted in all eight animals. Four of these animals had two transplants.
After a week, 1000 units of interferon solution prepared by the method of Interferon Production Example 2 was intraperitoneally injected twice a week for a total of 8 times.
The animals were sacrificed on the 42nd day and the fresh weight of the resulting tumor was measured. The remaining 4 mice were raised in the same manner as the previous 4 mice, except that interferon was not injected as a control, and they were sacrificed on the 42nd day after transplantation, and the fresh weight of the resulting tumor mass was measured. The measured results are shown in Table 4 below.

[Table] As is clear from the results in Tables 3 and 4, in those injected with interferon, the development of tumors was inhibited, and even if tumors did develop, their weight was lower than that of the control. extremely small,
It can be seen that its hypertrophy is significantly inhibited.
Nude mice injected with interferon also had better appetites and were more agile than control nude mice. Experiment 3 Acute toxicity test When we conducted an acute toxicity test of the interferon solution prepared by the method of Interferon Production Example 9 using 20-day-old mice, the toxicity of this interferon was extremely low, and it was found that the interferon was injected intraperitoneally. The LD ₅₀ at that time was found to be more than 20,000,000 units/Kg. As is clear from the above experiments, the interferon-sensitive diseases referred to in the present invention are diseases that are prevented or treated by the interferon used in the present invention, and are caused by viral diseases, such as Whether it is epidemic conjunctivitis, herpetic keratitis, influenza, rubella, serum hepatitis, etc., or non-viral diseases such as leukemia,
It may also be osteosarcoma. The form of the interferon-sensitive disease preventive or therapeutic agent containing interferon as an active ingredient of the present invention can be freely selected depending on the purpose. Examples include liquids such as sprays, eye drops, nasal drops, gargles, and injections, pastes such as ointments, and solids such as powders, granules, and tablets. These interferon-sensitive disease preventive agents,
The therapeutic agent should generally contain interferon with an activity of about 1 to 10,000,000 units per gram, and if necessary, other ingredients such as a therapeutic agent, an adjuvant, a filler, a stabilizer, etc. Alternatively, it is also possible to freely use two or more types together. In particular, as an injection, when administered intravenously all at once, interferon disappears from the blood in a short period of time and is easily excreted from the body. By using a method of intravenous drip infusion and extending the administration time, it is possible to effectively utilize the medicinal effects of interferon, and to prevent interferon-sensitive diseases by administering interferon.
You are also free to further enhance the effectiveness of your treatment. Two to three embodiments of the present invention will be described below. Example 1 Liquid Preparation A liquid preparation was prepared by adding 500 units of interferon per ml of interferon prepared by the method of Interferon Production Example 1 to physiological saline. This product is particularly suitable for spraying, eye drops, nasal drops, and gargling as a preventive or therapeutic agent for viral diseases such as epidemic conjunctivitis and influenza. Example 2 Injection Interferon prepared by the method of Interferon Production Example 9 was added to physiological saline at 100,000 g/ml.
An injection was prepared by containing the unit. This product is suitable for the prevention and treatment of interferon-sensitive diseases in general, such as viral diseases and tumor diseases. Example 3 Injection Interferon prepared by the method of Interferon Production Example 5 in 500 ml of 10% maltose solution
A drip injection was prepared containing 1,000,000 units and 100 mg of cyclophosphamide. This product is particularly suitable as a continuous administration injection of interferon for the prevention and treatment of tumor diseases. Example 4 Injection Interferon prepared by the method of Interferon Production Example 6 in 100 ml of 10% maltose solution
An injection was prepared containing 500,000 units of mitomycin C and 2 mg of mitomycin C. This product is particularly suitable for the prevention and treatment of neoplastic diseases. Example 5 Ointment When producing an ointment by mixing interferon powder prepared by the method of Interferon Production Example 4 with liquid paraffin and petrolatum according to a conventional method, interferon was added at a concentration of 10,000 g per 1 gram of the product.
It was contained so that it became a unit. This product is suitable for treatment of viral skin diseases, etc. Example 6 Tablets When tabletting interferon powder prepared by the method of Interferon Production Example 7 using a mixture of starch and maltose according to a conventional method, interferon was adjusted to 1000 units per tablet (100 mg) of the product. Tablets were manufactured by containing the following: This product is
Suitable for prevention and treatment of viral diseases of the digestive system. Example 7 Solution Interferon prepared by the method of Interferon Production Example 8 in 10 ml of 10% maltose solution.
A liquid preparation for internal use was prepared containing 200,000 units and 5 mg of methotrexate. This product is particularly suitable for the prevention and treatment of neoplastic diseases.

Claims (1)

[Scope of Claims] 1. Cultured human-derived cells are transplanted into the body of a warm-blooded animal other than humans, or the temperature of the cells is maintained within a diffusion chamber attached to or outside the body of a warm-blooded animal other than humans. Contains purified and fractionated interferon as an active ingredient, which is produced by multiplying the human-derived cells while being supplied with blood from animal body fluids, and then reacting with an interferon inducer in vivo or in vitro to produce interferon. An anti-interferon-sensitive disease agent characterized by: 2. The anti-interferon-sensitive disease agent according to claim 1, wherein the anti-interferon-sensitive disease agent is an agent for preventing interferon-sensitive diseases. 3. The anti-interferon-sensitive disease agent according to claim 1, wherein the anti-interferon-sensitive disease agent is a therapeutic agent for interferon-sensitive diseases. 4. The anti-interferon-sensitive disease agent according to claim 1, 2, or 3, wherein the interferon-sensitive disease is a tumor disease. 5 Interferon 1-10000000 per gram
The anti-interferon-sensitive disease agent according to claim 1, 2, 3, or 4, which contains a unit. 6 The anti-interferon-sensitive disease agent is a liquid preparation,
The anti-interferon-sensitive disease agent according to claim 1, 2, 3, 4, or 5, which is a base agent or a solid agent. 7 Claims 1 and 2 containing one or more of therapeutic agents, adjuvants, bulking agents, and stabilizers other than interferon;
The anti-interferon-sensitive disease agent according to item 3, 4, 5, or 6.