JPH08188539A - Method for alleviating pain symptom accompanied with carcinoma - Google Patents

Abstract

PURPOSE: To provide an immunotherapy capable of alleviating pains associated with malignant neoplasia especially of animal except human and treating diseases caused by virus by administering a composition containing a gonadotropin derivative and an immunostimulator. CONSTITUTION: A composition containing a substance composed of a gonadotropin (e.g. chorionic gonadotropin) or its derivative in an amount less than the lowest amount necessary to provoke a humoral immune response, as exemplified by the existence of a positive wheal upon subcutaneous injection and an immunostimulator (e.g. solubilized substance of Staphylococcus aureus) is administered. The composition is effective for treating cat leukemia, bovine leukemia, AIDS, etc., and for alleviating pain symptoms associated with malignant neoplasia and a dose is administered by subcutaneous injection once daily.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates generally to immunotherapy for the alleviation of symptoms of malignant tumors and the treatment of viral disease. More specifically, the invention relates to immunotherapy useful in the treatment of disease states such as feline leukemia, bovine leukemia and acquired immunodeficiency disease (AIDS). More specifically, the present invention relates to immunotherapy useful in alleviating pain associated with malignancy.

[0002]

2. Description of the Related Art In order to maintain the health of living organisms, advanced vertebrates have a complicated immune system that distinguishes foreign substances from "self" substances, Foreign substances are eliminated only after eliciting an immune response,
The self substance is what the living body allows. It is known that the mechanism for discriminating between self substances and foreign substances involves interactions between various leukocytes.

When an antigen is exposed to the circulating fluid of a living body, a substance (antigen) recognized by the immune system comes into contact with white blood cells of a type called macrophage. Macrophages are phagocytes and are therefore capable of phagocytosing and destroying substances that are not protected from the immune system by size, surface tissue (ie, smoothness), surface charge, or other mechanism.

Once phagocytosed by macrophages,
Upon treatment, the antigen or a portion thereof is deposited on the surface of macrophages and contacts thymocytes or another type of white blood cell called T cells. T cells control the production of antibodies by yet another type of white blood cell called B cells.

Antibodies are B cell-produced proteins and have the ability to bind to an antigen in a reaction specific to that antigen. Since an antibody binds only to certain parts of the surface of the antigen (antigenic determinants), the specificity of this antibody is determined by the extent to which the determinant to which it binds is not found on other antigens.

The binding of an antibody and its corresponding antigen on the surface of a foreign cell has the important consequences associated with the destruction of that cell by the immune system.

First, antibody coating of cells promotes phagocytosis of cells by macrophages and other types of phagocytic cells such as killer (K) cells and polymorphonuclear (PMN) spheres.
Killer cells have the effect of destroying antibody-coated cells, but need not be pre-exposed to sensitize macrophage-treated antigens.

Second, cell coating with antibodies activates a protein system known as the complement system in the liquid (plasma) fraction of blood. When this system is activated, complement components also coat foreign cells and promote phagocytosis. In addition, complement activation stimulates inflammatory cells, resulting in the production of chemicals that attract macrophages through a process called chemotaxis, which in turn causes inflammatory hormonal activation of cell function.

Finally, the complement component directly decomposes (lyses) the foreign cell membrane. The immune response associated with antigen-antibody and complement interactions is commonly referred to as the humoral response.

There are several types of T cells that regulate the humoral response. The types of T cells, helper (T _H) cells, inducer (T _I) cells, regulator (T _R)
Cells, and suppressor (T _S ) cells are described. [Herscowitz, Immunology III, Chapter 7, B
ellanti, JW Saunders, Philadelphia (1985)].
T _I and T _H cells are activated by contact with processed antigens on the surface of macrophages. Furthermore, T
_H cells are activated by signals from T _I and T _R cells. T _R cells are activated by signals from T _I and T _S cells. Activation of T _S cells occurs in response to signals from T _R cells or as a result of contact with antigen.

When an optimal amount of foreign substance is introduced into body fluid, B
The process by which cells produce antibodies is initiated. In this process, B cells are responsive to stimulation by T _H cells, the T _H cells are stimulated by macrophages earlier. The antibody production is because the signal from _TH cells to B cells is blocked by T _S cells when a few antigens are continuously introduced at a low level or when one antigen is introduced at a high level. Low levels or lack of production occurs. This block, called suppression,
It can be triggered through the macrophage-T _I -T _R pathway or by direct stimulation of T _S cells by antigen. Suppression of antibody production against the initial antigen can be overcome by a process known as anti-suppression. In this process, a second antigen whose antigenicity resembles that of the first, stimulates a subtype of T _S cells called anti-suppressor cells. These anti-suppressor cells
T _R cells send a signal, T _R cells, suppressor T _S
It confers resistance to cell activity on T _H cells and also blocks suppressor signals on suppressor T _S cells.
Gershon et al., J. Exp. Med., 153,1533-1546 (1981); Ya
mauchi et al., J. Exp. Med., 153, 1547-1561 (1981); and Green et al., Ann. Rev. Immunol., 1, 439-463 (1983).
Please refer to.

It is the action of T _H helper cells and T _S that determine whether an immune response occurs in the presence of an antigen.
Equilibrium of the action of suppressor cells. Therefore, in practice, the function of the T cell network could be examined with respect to the ratio of helper cells to suppressor cells (T _H / T _S ).

In addition, T cells are involved in another type of immune response that is said to be involved in cell-mediated immune (CMI) responses. T
When _H cells are contacted with the antigen that has been treated by macrophages, T _H cells release interleukin II (IL-2). This interleukin II (IL-2) is cytotoxic (T
_CYT) T cells were activated, further with gamma interferon released by T _H cells at this time, activates natural killer (NK) cells. T _CYT cells and
Both NK cells kill foreign cells. T _CYT is particularly involved in tumor cell rejection and destruction.

As is clear from the above discussion, an outline of the function of the immune system has already been obtained.

However, many of the functions of the immune system have not yet been elucidated. One of the unexplained areas is certain cancers (malignant tumors) and certain viruses [eg, feline leukemia virus, bovine leukemia virus, and human T-cell leukemia / lymphoma virus, which is considered to be the cause of AIDS.
(HTLV)] cells cannot be recognized by the immune system.

[0016] In order to stimulate the immune system against malignant tumors, many attempts have been made to enhance antitumor protection by administering an adjuvant (immunostimulator or enhancer).

These methods are based on macrophages and T
The purpose is to promote non-specific phagocytosis by cells and tumor cell death. Such methods can be used for infectious BCG mycobacteria, inactive Corynebacterium par
vum, glucan (glucose polymer obtained from microorganism),
Alternatively, levanizole, an anthelmintic known to be useful in stimulating CMI and macrophage activity, is used. Herberman et al., Immunology I
II, Chapter 19, (Bellanti), WB Saunders Co. (198
5), p. 343. Anti-tumor activity reported to have "lysosomal and pepsin lysates" containing glycoproteins from the cell wall of Lactobacillus bulgaricus [Bogdanov
Et al, FEBS Letters, 57: 259 (1975); Bogdanov et al, Byul.
letin Eksperimental 'noi Biologia i Meditsiiny, 84
: 709 (1977)], and destroyed Staphylococcus aur
Treatment of malignant tumors with eus [summary of Japanese Patent Application No. 84046487, which has been examined] is considered to belong to this category. The results of adjuvant therapy have various questions and limitations. Herberman et al., Supra.

The failure of the immune system to recognize malignant tumors
This is a particularly difficult problem considering the presence of above-standard levels of certain characteristic substances (tumor markers) in patients with various tumor diseases. Specifically, α
Fetoprotein (AFP), carcinoembryonic antigen (CEA), and human chorionic gonadotropin (HCG) are widely used fetal tumor markers for the examination of patients with liver, colon and trophoblastic tumors, respectively. 50 of patients with yolk sac tumor, hepatoma, retinoblastoma, embryonal carcinoma, breast and cervical cancer
% Or more of AFP above normal levels, and
Pancreatic cancer, melanoma, gastric cancer, basal cell cancer, bronchial cancer, pancreatic cancer, medullary thyroid cancer, familial medullary thyroid cancer, osteosarcoma, retinoblastoma, ovarian cystadenocarcinoma, fungal mycosis, hepatoma, High levels of AFP are found in 2 to 50% of patients with esophageal cancer, cervical adenocarcinoma, lung cancer, small bowel cancer, bladder cancer, and renal cell carcinoma,
In addition, neural crest tumor, breast cancer, prostate cancer, primary uveal cancer,
Neuroblastoma, malignant fluid, seminoma, basal cell carcinoma, gastric cancer, laryngeal cancer, endometrial cancer, cervical intraepithelial carcinoma, buccal mucosa cancer, craniopharyngioma, fetal rhabdomyosarcoma, oropharyngeal cancer , High levels of AFP are found in 9 to 50% of patients with brain tumors and testicular teratomas. HCG is found at high levels in the serum of more than 50% of patients with choriocarcinoma, malignant stromal cell tumor of the testis, nontesticular seminoma of the testis, embryonal carcinoma and pancreatic cancer, plus teratomas, ovarian adenocarcinoma,
It is found at high levels in 6 to 50% of patients with cervical cancer, endometrial cancer, seminoma, gastric cancer, bladder cancer, breast cancer, colorectal cancer, bronchial squamous cell carcinoma, melanoma, and multiple myeloma.
Tissue polypeptide antigen (TPA) and other universal embryonal tumor markers are also known and are associated with cell proliferation and are not specific to any species. Klavi
Anns of Clinical and Laboratory Science, 13
: See 275-280 (1983). Regarding HCG, Acevedo et al., Infection and Immunity, 31, 487-49
4 (1981), a chorionic gonadotropin-like antigen was found in bacteria isolated from the urine of cancer patients, but with the same bacterial species from other sources tested. Has not been found in. Furthermore, it was revealed that rat mammary adenocarcinoma cells and rat hepatoma cells synthesize villous gonadotropin-like substances, but Kellen et al.
As shown by Cancer Immunol. Immunother., 13, 2-4 (1982), this substance was not found in the serum of animals with these tumors.

Kellen et al. And US Pat. No. 4,384,
In 995, a subunit of HCG conjugated to a tetanus toxin was used to repeatedly inject this conjugate before exposure to tumor cells known to produce villous gonadotropin-like antigens. Thereby prophylactically stimulate the immune response to villous gonadotropin-like substances.

There are various differences between preventive therapy using HCG and adjuvant therapy. One of them is the same B cell that produces a specific antibody against a tumor antigen when eliciting an immune response for prevention. It is necessary to make repeated injections over a period of time in order to initiate the development of at least one population of (clones) and also for the antibodies produced by the clones. On the other hand, adjuvant therapy may cause antibody production by preexisting clones of B cells, and thus has an antitumor effect that can be immediately observed. Treatment with HCG conjugated to tetanus toxin (ie, treatment after the development of a malignant tumor) increases the likelihood of an uncontrolled Herxheimer-type reaction. The Herxheimer reaction occurs after treating a patient with syphilis with a substance that is toxic to Spirochetes, which causes syphilis, and is a potentially lethal toxic substance due to the large number of deaths of the bacterium. Released into the patient's bloodstream. By analogy, the cancer cells may die in large numbers and the patient may die at a previously unpredictable point in the process of eliciting an immune response to a tumor antigen.

Luteinizing hormone releasing factor (LHRF), often commonly referred to as gonadorelin, which releases luteinizing hormone, one of the pituitary gonadotropins, from the pituitary gland is described in US Pat. No. 4,002,738 and 4,071,6
In No. 22, it is used to treat various tumors. In addition, gonadorelin is used in the treatment of benign prostatic hyperplasia, a type of non-malignant but excessive prostate growth, in US Pat. No. 4,321,260. However, there is no suggestion in these patents that direct administration of gonadotropins may affect the destruction of malignant tumors. Furthermore, the release of luteinizing hormone from the pituitary is subject to feedback control that is independent of the administered gonadotropin, so even if luteinizing hormone is released, the amount released is merely knowledge of the dose. Can't decide. Moreover, LHRF, along with other substances, can act directly on tumor cells to increase villous gonadotropin secretion. By this, LH
The effects of RF administration are even more uncertain. Kellen
Et al, AACR Abstracts, 23, 235 (March 1982) (Summary 92
8).

In fact, Simon et al., JMCI, 70,
839-845 (1983) show that administration of gonadotropins and steroid hormones stimulates the growth of differentiated cancers. Such hormones include human follicle stimulating hormone (FSH), HCG, and human luteinizing hormone (LH).
H), and cortisol.

Thus, Simon et al. Seems to support the idea that direct administration of gonadotropins or steroid hormones has a proliferative effect on malignant tumors.

Evidence of suppression of the immune response to tumor cell antigens has been demonstrated by Akiyama et al., J. Immunol., 131, 3085-3090 (198).
It is shown in 3). According to it, the response of mixed cultures of lymphocytes taken from cancer patients and healthy donors was suppressed by introducing tumor cells taken from cancer patients into this system. This is because the not so the response of the cultures containing only lymphocytes taken from healthy donors inhibiting, in lymphocytes of cancer patients, the T _s cells exhibiting specificity for cells from tumor Suggests that it exists.

Furthermore, antigen-specific T _S cells have been isolated from mice with plasmacytoma, which cells blocked the in vitro induction of cytotoxic T cell responses against this tumor. Kolsch, Scand.J. Immunol., 19, 387-393 (19
84). According to Kolsch, T _S cells can be activated by high or low doses of antigen, and
It is possible to suppress the T _H cells, the use of the critical or intermediate antigen dose which activates T _H cells at the same time as activating T _S cells, T _H cells may become dominant it can. Thus, Kolsch suggests that T _H cells are activated, but at this time there is an antigen dose that achieves the delicate balance of T _S cells governing the immune response.

Loblay et al., Aust. J. Exp. Biol. Med. Sc
i., 62, 11-25 (1984) suggest that the suppression caused by T _S cells in animals exposed to an antigen is enhanced by subsequent administration of a sufficient amount of antigen. There is. It is therefore surprising that attempts to induce anti-suppression were aimed at supplying anti-suppressor cells, or substances derived therefrom, rather than directly inducing anti-suppression.

Green, “Antisuppression: The Potential Role of T-Cells In Cancer Therap
y) '', Fefer et al., Raven Press, New York (1982), and Green et al., Ann. Rev. Immunol., 1, 439-463 (198).
See 3).

[0028]

Accordingly, the method of the present invention for alleviating the painful symptoms associated with malignant tumors induces a humoral immune response, exemplified by the evaluation by positive wheal following subcutaneous injection. Administering to a patient / animal in an amount less than the minimum amount necessary for the treatment, selected from the group consisting of a substance specific to diseased cells of a malignant tumor / patient, an effective fragment thereof, and an effective derivative thereof. It includes a process. As an example of such a method, HCG and Staphyl
There is once daily administration of a composition containing a lysate of ococcus aureus. Methods for alleviating the symptoms of acquired immunodeficiency in a patient / animal in accordance with the present invention are described in more than the minimum amount required to elicit a humoral immune response, as exemplified by the positive wheal assessment following subcutaneous injection. Administering a small amount of a substance selected from the group consisting of a substance specific to diseased cells of an acquired immunodeficiency patient / patient, an effective fragment thereof and an effective derivative thereof to the patient / animal.

A composition for alleviating pain symptoms associated with a malignant tumor in a patient / animal according to the present invention comprises less than the minimum amount required to elicit a humoral immune response, the diseased cells in the malignant tumor / animal. The substance selected from the group consisting of a substance peculiar to the above, an effective fragment thereof, and an effective derivative thereof.

The mixture according to the invention further comprises an immunostimulant in an amount less than the minimum required to elicit a humoral immune response, as exemplified by the evaluation by positive wheal after subcutaneous injection. As an example of such a composition,
There is a mixture containing lysates of HCG and Staphylococcus aureus.

Another composition according to the invention for alleviating the symptoms of acquired immunodeficiency in a patient / animal is the use of an amount of acquired immunity less than the minimum required to elicit a humoral immune response. It comprises a substance selected from the group consisting of substances specific to diseased cells of deficient patients / animals, effective fragments thereof and effective derivatives thereof. The composition further comprises an amount of an immunostimulatory agent less than the minimum required to elicit a humoral immune response, as exemplified by the evaluation of positive wheal following subcutaneous injection. As an example of such a composition,
There is a mixture containing lysates of HCG and Staphylococcus aureus.

In one method according to the present invention, a malignant tumor or a substance specific to AIDS is added to malignant tumor or
It is administered to patients / animals with AIDS in doses believed to be less than the minimum amount required to elicit a humoral immune response (ie, initiate antibody production). This dose is administered daily until the symptoms of the disease disappear, but it may be continued for longer if desired, and there is no problem.

To determine doses lower than those required to elicit a humoral immune response, Moore, Clinic
Wheal caused by subcutaneous injection of therapeutic agent is evaluated according to the procedure described in al Medicine, 81, 16-19 (1974). According to Moore's paper, such assessments are used to determine doses of vaccines useful in eradicating the symptoms of influenza. Subcutaneous injection takes 10 minutes after the injection and is white, firm, stepped, and disc-shaped (like a disc adhered to the skin, well-balanced and distinct from the circumference) ) Wheals are determined to be positive. A negative wheal indicates that the dose is below the dose required to elicit an immune response, is absorbed after 10 minutes and is softer and flatter than at injection.

This negative wheal is jagged with uneven edges and has a diameter of less than 2 mm on average.

The preferred dose, 2 International Units (IU), is
Initially determined by the equine skin test of Example 1, the following examples show that treatment can often be successful by administering a dose level of 2 IU. The dose of 2 IU appeared to be sufficiently low for both animals and humans tested, without first determining the optimal dose by skin testing. However, one of ordinary skill in the art would appreciate that
Alternatively, using appropriate dosage determination by another method,
It is considered possible to refine the method of the invention.
Bacterial lysates serve as broad-spectrum stimulators of cell-mediated immunity to complement the action of villous gonadotropins and simultaneously prevent the toxic response induced by rapid scabbing of necrotic tissue similar to the Helxheimer response , Added to each treatment vial. There is no species specificity in the response,
Humans respond to equine chorionic gonadotropin (ECG) or HCG, and other animals in the examples also show ECG or HC
Responded well to treatment with G.

It has been reported that AIDS patients have a greatly reduced ratio of T _H cells to T _S cells. Cohen, B
ritish Journal of Hospital Medicine, 31, 250-259
(1984). AIDS is the Centers for D
It is defined by isease Control as a disease that, at least to some extent, suggests a deficiency in cell-mediated immunity that occurs in people who have no known cause for decreased resistance to the disease.

The virus considered to be the cause of AIDS is a kind of human T-cell leukemia / lymphoma virus (HTLV), which is known as HTLV-III virus, and the virus is feline and bovine leukemia. It has been reported to be related to the virus that causes
Franklin, Science News, 126, 269 (1984). Feline and bovine leukemia viruses are known to be similar in antigenicity. Morgan et al., J. Virol., 46, 177-186.
(1983). Therefore, the present inventors' findings that administration of HCG in an amount less than that required to elicit a humoral immune response may help alleviate the symptoms of feline and bovine leukemias. AI is similar treatment
It suggests that it may be effective in alleviating the symptoms of DS.

Furthermore, feline leukemia virus (FLV) and
Both HTLVs are retroviruses (type C virus, RNA
(Also known as tumor virus, leukemia virus)
Is. Manzani et al., Surv. Immunol. Res., 1, 122-
125 (1982). Retroviruses can be transmitted as infectious particles containing viral genes encoded in the form of ribonucleic acid (RNA). In infected cells, this RNA is encoded in deoxyribonucleic acid (DNA) by a viral enzyme called reverse transcriptase.
The viral gene loaded in the DNA is the DNA of the infected cell.
It is integrated with the genetic material encoded by, and is replicated, transcribed and translated. Lewin, Genes, Chapter 13, John Wiley
andSons, New York (1983). Such retroviruses generally produce a steady state of infection, with the progeny of the virus being continuously shed by sprouting from the surface of the host cell. In this way, steady-state viruses represent a clinical criterion for induction of resistance. High doses of inoculum of viral antigens, persistence of virus-specific antigens, and production and persistence of tumor-specific antigens. Herberman etc. mentioned above. Thus, the methods and compositions of the present invention shown in the examples below to be effective in the treatment of various tumor diseases, particularly feline leukemia and bovine leukemia, are also believed to be effective in the treatment of AIDS. .

[0039]

EXAMPLES Examples 1, 2 and 3 relate to the treatment of horses with two different malignancies. Examples 4 and 5
Relates to the treatment of cats diagnosed with cat leukemia. Example 6 relates to the treatment of cows with bovine leukemia. Examples 7 and 8 relate to the treatment of dogs with two different malignancies. Example 9 relates to the treatment of patients with malignant tumors. Example 10 relates to the pain relief effect of treating patients with malignant tumors.

Example 1 Horses with mastocytomas were treated daily with 2 IU of gonadotropin alone (ie, without the bacterial lysate immunostimulant). The gonadotropin used is WA
Butler Company's equine villus gonadotropin, or
It was human chorionic gonadotropin from Ayerst Corp. (APL-human chorionic gonadotropin). In this and the following examples, 2 IU of horse chorionic gonadotropin daily
(ECG) or HCG was administered. By the time the horse died of the Herxheimer reaction, the horse's tumor shrank rapidly and significantly. (No such side effect was observed in animals treated with other immunostimulants.) Example 2 Three horses with melanoma were treated by the procedure of Example 1 until the symptoms disappeared. Successfully treated. However, a bacterial lysate immunostimulator was added.

After the symptom disappeared, the three did not receive any further treatment, but the symptom did not recur.

A bacterial lysate suitable for use in this procedure is Delmont Lasate under the name Staphage Lysate®.
It is on sale from bs. Staphage Lysate® is a bacteriologically sterilized staphylococcal vaccine that contains Staphylococcus aureus components and media components (sodium chloride and ultrafiltered beef heart infusion broth). Staphylo is a component of Staphylococcus
Mother cultures of coccus aureus serology type II and III are prepared by lysis with polyvalent staphylococcal bacteriophage. 120 ml to 180 million Staphylococcus aureus colony forming units and 100 million to 1 staphylococcal bacteriophage plaque forming units per ml.
Including 100 million. All test agents used for treatment were 0.5 c each
Injectable drug c, gonadotropin 2 units and Staphag
Contains 0.1 cc of Lysate®.

Regardless of the type of drug administered , the type of malignancy, or the size of the tumor,
All animals will lose their tumor or leukemia until
Subcutaneous injections of villous gonadotropin and immunostimulant mixture were received once daily.

Tests and Results All animals were diagnosed with a tumor by a licensed veterinarian.

Example 3 One horse having an undifferentiated tumor on the face was successfully treated by the procedure of Example 2 until the symptoms disappeared. After the symptoms disappeared, the horse received no further treatment, but the symptoms did not recur.

Example 4 A double blind study of the treatment of cats diagnosed as infected with feline leukemia virus was performed by a licensed testing laboratory. Feline leukemia clinical trials were performed by a licensed laboratory, and biopsy tissues taken from some of the animals were examined at a veterinary commissioned laboratory. In all cases, the animals were treated as above and treatment was discontinued when the symptoms disappeared. There was no need to perform a second treatment in any case. No side effects were seen.

All animals receiving placebo died within 7 days. Some of the treated animals survived after 6 weeks and had not been treated since the experiment.
Of particular interest was that the treated cat was not only asymptomatic, but not viremia. In any case, six of the eight animals treated (cats numbered 2,3,11,21,23 and 24) had a survival time as compared to animals receiving placebo. Prolonged or alleviated symptoms.

Cats numbered 4, 5, 16 and 22 received placebo. Cat # 4 died one day after the start of the study. No cat # 4 was necropsied.

No. 5 cat did not show much change in viremia. Tumor size remained constant. The cat was euthanized 10 days after the start of the test.

Upon autopsy, it was confirmed that the cat was moribund, diminished, blind, and anemic. Tumors were in the tongue, eyes, pleural, perirenal and peritoneal cavities.

The cat number 16 died 6 days after the first treatment. At necropsy, small metastatic tumors were found in the lungs and larger tumors were found in the retina.

Cat # 22 died 6 days after initial treatment. At necropsy, metastatic tumors were found in lung, liver and retina. The cause of death was renal hemorrhage to the retroperitoneum below the lumbar region.

Numbers 2, 3, 7, 11, 15, 21, 23 and 24
The cat received experimental treatment.

In cat # 2, viremia was reduced but tumor size did not change. Cat # 2 died 16 days after the start of the study. Autopsy revealed a tumor, which was open and shed. Systemic lymph node enlargement, hepatic hypertrophy, and small tumor in the apex of the heart were observed. In cat # 3, viremia was reduced and became negative. The tumor had decreased in size and returned to normal. During the test period,
The cat was normal in appearance.

Cat # 7 died 4 days after the first treatment. No necropsy was done.

Viremia was slightly reduced in cat # 11. Tumor size did not change. Number 11
The cat died 19 days after first treatment. Upon autopsy, a tumor was found throughout the body cavity.

In cat # 15, viremia was not significantly reduced. No change was seen in the tumor. The cat died 10 days after first treatment. At necropsy, the cat was thin and lean, with a tumor
It was found in the mediastinum, pericardium, pleura, and illiac lymph nodes.

In cat No. 21, viremia was unchanged. This cat appeared normal. At necropsy, thymus enlargement and mesenteric lymph node enlargement were observed.

In cat # 23, viremia was reduced throughout the study.

The cat number 23 remained normal for the duration of the test. In cat number 24, viremia was unchanged. This cat is
Seemed normal. At necropsy, the thymus was enlarged, but the cat was otherwise normal.

Example 5 A few dozen cats were treated for feline leukemia by the procedure of Example 2 by veterinarians in Ohio, Pennsylvania and North Carolina, and treatment was successful.
Based on the reported findings of the treating veterinarian, at least 80
% Recovered health after cessation of treatment. For example, one cat had just barely lifted his chest wall to breathe, reducing his weight from 13 pounds to 6 pounds. After 10 days of treatment, the cat roamed and became active. Weight increased to 10 pounds.

Example 6 As a result of treating 20 cases of bovine leukemia by the procedure of Example 2, the symptoms were completely ameliorated over a long period of 13 months.
Milk production in 17 dairy cows not only recovered, but improved. Only 3 cases failed treatment,
In these cases, the cows were moribund several days before the start of treatment. Clinical trials of bovine leukemia were conducted by a licensed laboratory and biopsy tissues of some cases were examined by a veterinary contract laboratory.

Example 7 Squamous cell carcinoma of the jaw of a 6 month old puppy was treated by the procedure of Example 2 above. According to an evaluation by a veterinary contract laboratory that examined the biopsy tissue, "the prognosis was not good", but as the continuous X-ray record shows,
This dog has completely healed.

Squamous cell carcinoma of the shoulder of a 12 year old dog was treated by the procedure of Example 2 above. Two days later, the dog was able to walk longer, had more food, and the tumor felt warm to the touch. It was observed that the tumor had clearly shrunk after 5 days. Two
After a week, the tumor had almost disappeared.

Example 8 A 13-year-old dog develops anal tumors that grow in size day by day,
The tail was pushed away laterally. The dog was treated according to the procedure of Example 2. Photographic records showed that the tumor size began to decrease from day 3 and became smaller day by day. The tumor disappeared. The dog died later, but the cause is unknown. No necropsy was done.

Example 9 A small number of patients diagnosed with end-stage cancer including melanoma, colon cancer, breast cancer, liver cancer, pancreatic cancer and lung cancer were treated.
The patient was treated according to the procedure of Example 2, provided that
Some patients received 0.2 cc of immunostimulant per unit dose.

Nine patients treated at one location for a period of two and a half years all survived longer than the expected survival time indicated by the physician treating the patient, with only one dying. Patients who died discontinued the treatment according to the invention 2 months before their death. Of the 8 surviving patients, 6 will have been living for a year and a half or more since starting treatment.

All nine of these patients practiced the invention as a last resort. And, with the exception of one patient who died, all the remaining patients had received some radiation or chemotherapy prior to initiating the treatment according to the invention. All 9 patients had metastases, indicating that the disease was progressing. No side effects were seen.

However, it is not clear to what extent all patients regularly used the mixture according to the invention. For example, some surviving patients have reduced the frequency of treatment to once or twice a week, but have not reduced their overall health. None of these patients, with the exception of those who died, completely discontinued treatment.

Sporadic treatments elsewhere have been less successful, and overall, approximately half of the treated patients are still alive. The other half of the patients died. With no autopsy data, it is unclear whether all causes of death in treated patients could be cancer or other causes. Patients who did not receive maximal chemotherapy and radiation therapy responded most clearly to the mixtures and methods of the invention. This indicates that the therapeutic agents of the invention are, at least in part, associated with immune manipulation, and that the cells responsible for the immune response of patients undergoing conventional anti-cancer therapy are weakened. Suggesting that

Example 10 Many of the patients mentioned in Example 9 and other patients treated according to the invention for various cancers reported that most or all of the pain associated with these cancers was reduced. did. Despite the treatment according to the invention, the level of pain suffered by the patient was also considerably reduced in most cases, even in the case of the patient who died soon.

In many cases, treatment with anesthetics and other analgesics could be discontinued, resulting in patient relief from drug-induced apathy and mental distress.

Patients were treated according to the procedure of Example 2, with the exception that some patients received 0.2 cc of immunostimulant per unit dose. However, it is not clear to what extent all patients regularly used the composition according to the invention. Some patients receive 1 treatment daily
It is thought that the degree was reduced to once or twice a week.

Patients who experienced pain due to a disease state due to a malignant tumor reported the following results.

Patient A was a 74 year old boy with cancer of the spine, which had spread to the lungs. The patient reported suffering a significant amount of pain and required 4-6 doses of Perkoset per day. Although this drug only slightly helped reduce pain, it caused mental distress and apathy. Within 9 days of starting the treatment according to the invention, the patient reported no pain.

The patient was relieved of pain for over 4 months, but the treatment according to the invention was discontinued because the patient had pneumonia. Treatment was resumed but no longer reported pain relief.

Patient B reportedly suffers from breast, lung and bone cancers and suffers from severe pain before the start of the treatment according to the invention. The treatment according to the invention did not completely eliminate the pain in the patient, but was reported to reduce the pain.

Patient C reported suffering from severe pain due to bone cancer, which pain was caused by treatment according to the invention.
It didn't reduce at all until it continued for a week. However,
At the end of this 5 week period, the patient reported that his pain had been significantly reduced.

Patient D reported suffering severe pain from cancer of the liver and pancreas. This pain is
It was reported that the treatment with the method of the present invention had almost disappeared after 2 weeks of treatment.

Patient E reported suffering severe pain from cancer of the breast, skin, lungs and lymph vessels.
After being treated with the method according to the invention for 7 weeks, the symptoms did not improve. So, when the dose of treatment was increased, the patient reported that the pain dramatically disappeared within 4 days.

Patient F reported suffering severe pain from lung cancer. After receiving 5-6 days of treatment according to the invention, the patient was able to discontinue the treatment of pain with codeine and morphine. It was then reported that the pain did not recur.

Patient G reported moderate pain due to lung cancer. It was reported that this pain disappeared as a result of treatment with the method of the invention.

Patient H reported uncontrollable pain due to cancer of the colon, lungs and bones. This pain did not respond to previous treatment with anesthetics and other analgesics. 7 from the start of the treatment according to the invention
Within days, the patient reported that the pain had disappeared.

Patient I had cancer of the liver and aorta and had mild discomfort, which he described as "pulling or getting caught." It was reported that this pain was not relieved by the method according to the invention.

Patient J, a 40 year old female, reported having suffered moderate pain due to visceral cancer of multiple organs. The patient died 4 weeks later, but the patient's pain was reported to have disappeared with the treatment according to the invention.

Patient K, a 50 year old boy, suffered severe pain from terminal lung cancer.

The patient died 2 weeks after the start of the treatment according to the invention, at which time the patient was reported to be relatively painless.

Patient L, a 70 year old male, reported suffering severe pain from colon cancer. The patient died 4 weeks after the start of the treatment according to the invention, but the patient reported relieving pain over the last 2-3 weeks.

Patient M, a 55 year old female, had breast cancer, and the cancer had also spread to her arms and shoulders.

The arm was extremely swollen and the skin was sore that it was sore that he could not sleep for 10 to 20 minutes or more at a time. It was reported that the treatments carried out according to the invention significantly reduced, if not completely, the pain after 2-3 weeks.

Patient N was a 70 year old male who reported suffering from migratory pain due to bone cancer. This pain was previously treatable, but was reported to have disappeared after 3 weeks of treatment with the method of the invention.

The efficacy of treatment of malignancy, cat leukemia, and AIDS with gonadotropins could be explained in several ways. The response achieved by the present invention is rapid even when HCG alone is administered as in Example 3. This rapidity suggests that the present invention does not work merely by initiating a humoral immune response. This can be explained as follows. That is, a small dose of gonadotropin stimulates the anti-inhibitory response by altering the T _H / T _S ratio balance to favor T _H cell activation. As a result, the existing immune response (including CMI response) to the disease is initiated. Other explanations are possible. That is, gonadotropins stop the production of gonadotropin-like molecules by malignant cells in a negative feedback mechanism. As a result, the surface charge associated with gonadotropin-like molecules is changed from a negative charge to a positive charge, which charge either promotes phagocytosis by macrophages or otherwise exposes hidden tumor antigens.

[0093]

While the present invention has been described in terms of particular methods and compositions, it will be apparent to those skilled in the art that various modifications and variations will be possible in light of the present disclosure.

For example, the various derivatives and fragments of the disclosed human chorionic gonadotrophin, as well as other tumor-specific or virus-specific antigens, are believed to be effective in accordance with the present invention. In addition, the preferred route of administration is subcutaneous injection, but intramuscular, intraperitoneal, or intravenous
Of course, there is no need to exclude intranasal or other effective routes of administration from the scope of the present invention.

In addition, other tumor markers, such as
Carcinoembryonic antigen, α-fetoprotein and tissue polypeptide antigens, along with HCG, have been classified as appearing with malignant tumors and are therefore likely to be equally effective. These substances are also included in the scope of the present invention.

Further, human chorionic gonadotropin, follicle-stimulating hormone, luteinizing hormone and thyroid-stimulating hormone are glycoproteins each consisting of one α subunit and one β subunit. The subunits of HCG differ only slightly from the same α subunits of FSH, LH, and TSH. Although the significance of the subunit structure has not yet been determined, α-HCG and β-
Both HCG are associated with tumors. Acevedo, Infe
See ction and Immunity, 31, 487-494 (1981). Therefore, these pituitary hormones are included within the scope of the present invention.

Furthermore, immunostimulants other than the lysate of Staphylococcus aureus are considered to show equivalent efficacy. Such other immunostimulants include, for example:
There are BCG mycobacteria, Corynebacterium Baruum and levamisole. Therefore, these immunostimulants are also included in the scope of the present invention.

It is believed that all effective fragments and effective derivatives of the substances suggested herein will be produced by those skilled in the art and used in accordance with the present invention. These fragments and derivatives are also within the scope of the invention as claimed.

Therefore, the invention described in the section of the claims should be construed to include the above-described modifications within the scope of equivalents thereof.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display area C07K 14/59 8318-4H C12N 1/20 F 8828-4B // (A61K 38/24 AAH 35: 74) (C12N 1/20 F C12R 1: 445) A61K 35:74)

Claims (14)

[Claims] 1. A method for alleviating pain symptoms associated with a malignant tumor in a non-human animal, which comprises eliciting a humoral immune response, which is exemplified by the following steps: expression of positive wheal after subcutaneous administration. A composition for alleviating pain symptoms associated with a malignant tumor, comprising a substance selected from the group consisting of gonadotropins, effective fragments thereof, and effective derivatives thereof in an amount less than the minimum amount required for Is administered to an animal other than human, the method for alleviating the pain symptom associated with a malignant tumor in an animal other than human. 2. The method of claim 2, wherein the gonadotropin is villous gonadotropin. 3. The immunostimulant is an immunostimulant in an amount smaller than the minimum amount required to elicit a humoral immune response together with gonadotropin, which is exemplified by the expression of positive wheal after subcutaneous administration. The method according to 1 or 2. 4. The method of claim 3, wherein the immunostimulant is a bacterial lysate. 5. The bacterial lysate is Staphylococcus aur
The method according to claim 4, which is a lysate of eus. 6. The method of claim 1, wherein the gonadotropin is a pituitary gonadotropin. 7. The method according to claim 6, wherein the pituitary gonadotropin is a substance selected from the group consisting of luteinizing hormone and follicle stimulating hormone. 8. The immunostimulator is an immunostimulator in an amount smaller than the minimum amount required to induce a humoral immune response together with gonadotropin, which is exemplified by the expression of positive wheal after subcutaneous administration. The method according to 6 or 7. 9. The method of claim 8, wherein the immunostimulant is a bacterial lysate. 10. The bacterial lysate is Staphylococcus a
The method according to claim 9, which is a lysate of ureus. 11. The method of claim 1, wherein the gonadotropin is thyroid stimulating hormone. 12. The immunostimulant is an immunostimulant in an amount smaller than the minimum amount required to elicit a humoral immune response together with gonadotropin, which is exemplified by the expression of positive wheal after subcutaneous administration. The method described in 11. 13. The method of claim 12, wherein the immunostimulant is a bacterial lysate. 14. The bacterial lysate is Staphylococcus a
14. The method according to claim 13, which is a lysate of ureus.