JPH06316529A - Pharmaceutical composition for treatment and prevention of opportunistic infection accompanying lentivirus infection - Google Patents

Abstract

PURPOSE:To provide a pharmaceutical composition effective for the treatment and prevention of opportunistic infection frequently occurring in an immunologically deficient state caused by lentivirus infection. CONSTITUTION:The pharmaceutical composition contains a protein belonging to unmodified transferrin-lactoferrin group. Opportunistic infection diseases frequently occurring in immunodeficiency caused by lentivirus infection can effectively be treated and prevented by administering the pharmaceutical composition by oral administration or topical application as an external agent. It can be administered absolutely without causing the problem of side action except for patient having cow milk allergy or chicken egg allergy.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a pharmaceutical composition containing native transferrin / lactoferrin group proteins for the treatment and prevention of opportunistic infections associated with lentivirus infection.

[0002]

2. Description of the Related Art Acquired I, an acquired immunodeficiency syndrome, which is one of the biggest problems in modern medicine
mmunodeficiency Syndrome;
AIDS) is an infectious disease of RNA virus called lentivirus. Lentivirus belongs to the retrovirus group, and when it infects a host cell, viral RNA is transcribed into DNA by the action of reverse transcriptase, and it behaves like a gene of the cell itself. Therefore, once the virus is infected, there are no cures in modern medicine, as changes in the gene level occur as in malignant tumors. That is why AIDS has become a fear of people.

The human lentiviruses, HIV-1 and HIV-2, mainly destroy lymphocytes having the CD4 receptor, and are therefore characterized by impaired immune function. The lentivirus group includes human HIV-1 and H
SI that causes immunodeficiency syndrome in monkeys other than IV-2
V, feline immunodeficiency syndrome virus (FIV), goat arthritis / encephalitis virus, equine infectious anemia virus, and bovine immunodeficiency syndrome virus are known. A common feature of lentivirus infections in humans, monkeys, cats, and cattle is an immunodeficiency and frequent opportunistic infections. In particular, in human AIDS, while suppressing viral growth by administration of antiviral agents, treatment and prevention of opportunistic infections include HIV carriers who are asymptomatic, AIDS-related syndromes, and patients with genuine AIDS who have a remaining quality of life ( qua
It is important for improving the life of life and extending the life.

[0004] Opportunistic infections refer to infections with low pathogenic microorganisms, which have not been a serious problem in the past.
The mechanism of its occurrence in AIDS is due to immunodeficiency due to a decrease in CD4 lymphocytes, and infection with various opportunistic pathogens. In AIDS patients, pneumonia caused by respiratory infection of protozoa, Pneumocystis carinii, cytomegalovirus (Cyto
megalovirus), Herpes simplex virus (Herpes simplex virus)
s), systemic infections such as Candida, and fungal infections such as Aspergillus, which are very characteristic opportunistic infections.

[Problems to be Solved by the Invention]

Despite the remarkable development of various antibiotics and synthetic chemotherapeutic agents (hereinafter collectively referred to as chemotherapeutic agents), opportunistic infections are known to be extremely intractable. The process by which animals treated with chemotherapeutic agents recover from infectious diseases plays a crucial role in the elimination of pathogens by the host's biological defense system, as well as the inhibition or killing of pathogens by chemotherapeutic agents. It has been known. On the other hand, opportunistic infections occur only in hosts with impaired host defenses, and because multiple pathogens are transmitted, a single chemotherapeutic agent is often not sufficient. . Therefore, it is natural that opportunistic infections are not completely cured unless the pathogenic growth of many pathogens is blocked by multiple chemotherapeutic agents and at the same time the host body's defense ability is improved by some means. Up to now, some methods have been known to improve the biological defense ability of the host, which has declined, but no methods have been known that can be carried out easily without any side effects. An object of the present invention is to provide a pharmaceutical composition for treating / preventing opportunistic infections that frequently occur in an immunodeficient state caused by lentivirus infection.

[0006]

Means for Solving the Problems The pharmaceutical composition provided by the present invention is characterized by containing a protein belonging to the native transferrin-lactoferrin group.
Although these proteins are widely distributed in animals, resources that are abundant and can be separated and purified economically include mammalian blood in the case of transferrin, mammalian milk in lactoferrin, and avian milk in ovotransferrin. It is egg white. Transferrin-lactoferrin group proteins should be separated and purified by a general method for separating and purifying proteins from these resources, that is, ultrafiltration, ion exchange chromatography, gel filtration chromatography, and fractional precipitation with ammonium sulfate. You can

Transferrin / lactoferrin group proteins are large molecules having a molecular weight of more than 70,000 daltons.
It has the characteristic of chelating individual ferric iron ions. Moreover, it is considered that transferrin-lactoferrin group proteins obtained from any of the animal species have large structural similarities, and thus proteins having the same origin are gradually differentiated in the process of evolution. Transferrin-lactoferrin group proteins isolated from nature have an iron content that reflects the existing state in nature. For example, the iron ion saturation of lactoferrin separated and purified from milk by repeating ion exchange chromatography and gel filtration chromatography is usually 25-28%.
It's about.

[0008] Iron-unsaturated natural transferrin / lactoferrin group proteins have a pH value in the presence of citric acid.
It can be converted to an apo type containing no iron ion by ultrafiltration at 3.0. Further, by adding a calculated amount of ferric iron ion to apo-type protein in the presence of carbonate ion, a protein chelated with ferric iron ion at an arbitrary saturation can be obtained. As the transferrin-lactoferrin group protein used in the present invention, a protein in which iron is saturated at any ratio from a trivalent iron ion-free apo type to a holo type in which iron ions are completely saturated is used. You can That is, the efficacy of the pharmaceutical composition of the present invention is independent of iron saturation.

[0009] The transferrin-lactoferrin group protein thus prepared can treat and prevent opportunistic infections in mammals, including humans, by improving the biological defense ability of mammals that has been reduced by lentivirus infection. . The oral route, which is the most common route for administering a drug, can sufficiently achieve the purpose of therapeutic and preventive measures. In the special case, in the case of opportunistic infections of the mouth, skin and eyes, it can be applied topically as an external preparation. Dosage interval is usually 1 day
It is sufficient to take the medicine once, for example, early in the morning on an empty stomach or before going to bed.

The transferrin-lactoferrin group protein has a weak antibacterial activity against a certain kind of microorganisms, and especially the apo-type antibacterial activity is enhanced on a medium in which iron is a growth limiting factor. However, since these proteins taken orally cannot be absorbed into the body from the intestinal tract while maintaining their molecular forms, their antibacterial activity, like chemotherapeutic agents, is a treatment for opportunistic infections. There is no possibility of being linked to preventive action. Its action is via the host's biological defense system. A characteristic of the host-mediated action of physiologically active proteins in vivo such as transferrin-lactoferrin group protein is that the dose / response curve exhibits a sigmoid type, and the response reaches a peak when the dose exceeds a certain level. Also in the case of transferrin-lactoferrin group protein, the dose-response curve is sigmoid type, and the dose at which the response peaks is in the range of 0.5-5 mg / kg by oral administration.

It is not yet completely known what kind of host-mediated reaction is induced by ingestion of transferrin-lactoferrin group proteins. Oral administration of these to small animals such as mice and rats enhances the foreign body phagocytic function of phagocytic cells present in peripheral blood, alveoli, and abdominal cavity, and thus certainly promotes the terminal stage of pathogen treatment. Is.

[0012]

[Action] It can be proved from some cases that the transferrin-lactoferrin group protein is remarkably effective in the treatment and prevention of opportunistic infections associated with lentivirus virus infection. For example, 600 mg / day of bovine lactoferrin to AIDS patients confirmed to be infected with HIV-1 by blood test.
Oral administration of orally daily decreases the frequency of opportunistic infections significantly, discontinuation of treatment causes recurrence of opportunistic infections, and transferrin is used in patients with carini pneumonia, the most opportunistic infection of AIDS. -Oral administration of lactoferrin group proteins markedly shortens the period leading to remission, and shortens the period of morbidity in viral infections such as cytomegalovirus and herpes simplex urs. . Patients who received the drug had an increased appetite, and their weight lost during the disease period recovered rapidly,
The effects of transferrin / lactoferrin group proteins are clear.

On the other hand, FIV of cats belonging to the lentivirus group, when it falls into an immunodeficient state after a latent period, causes opportunistic infections in the respiratory and urinary systems and dies early or later. Another notable symptom of FIV immunodeficiency in cats is the frequent occurrence of stomatitis.

The most common stomatitis in cats is aphthous stomatitis due to herpes simplex virus infection. At the time of initial infection, a high fever is observed for 1 to 2 days, followed by pain in the mouth, drooling, malodor in the mouth, and anorexia. This disease is a typical opportunistic infection that recures any number of times when the body's defense ability decreases. Lentivirus virus infection has been confirmed by antibody test in blood, and transferrin / lactoferrin group proteins are applied to cats with stomatitis at 10-20 mg / kg per day by topical application as an ointment or ingestion by mouth. After the fresh treatment, many ulcers in the mouth recovered and healed within a few days. Empirically, stomatitis due to FIV infection is difficult to recover because of severe immunodeficiency, and many individuals die because of their weakness. Therefore, the therapeutic effect of transferrin / lactoferrin group proteins seems to be remarkable.

Furthermore, when FIV-infected individuals were mixed with transferrin / lactoferrin group protein in the feed and given for a long period of time, opportunistic infections of respiratory and urinary system were clearly reduced. This fact means that the protein prevented the opportunistic pathogens from multiplying and developing infectious diseases through strengthening the immune system.

As described above, the transferrin-lactoferrin group proteins show a significant therapeutic effect on opportunistic infections associated with lentivirus virus infection by oral administration or local administration by external preparation. When given to individuals who have not developed an opportunistic infection, the frequency of infection can be significantly reduced.

Moreover, it is noteworthy that side effects associated with administration of transferrin / lactoferrin group proteins were hardly observed, irrespective of whether they were therapeutic or prophylactic. As an exception that side effects were observed, in the case of lactoferrin, milk allergy patients,
With ovotransferrin, allergic symptoms such as redness and itchiness on the skin of the whole body were only observed in chicken egg allergy patients. Therefore, this side effect can be avoided by giving lactoferrin purified from milk to hen egg allergy patients and ovotransferrin to milk allergy patients. Further, when a large amount of apo-type protein exceeding 20 grams was orally administered to an adult male at one time, diarrhea sometimes occurred. Transferrin
The lactoferrin group protein is resistant to proteolytic enzymes, and it is considered that this is a phenomenon caused by a large amount of this protein flowing into the lower digestive tract with water.

When the protein of the present invention is administered, the oral route is usually selected, but it can be applied as an external preparation for opportunistic infections of the oral mucosa, skin, eyes, throat and bronchi. The administration period should be at the time of prevention and treatment of opportunistic infections by lentivirus-infected individuals.

In carrying out the present invention, milk, skim milk, whey, whey protein and their powders containing native transferrin-lactoferrin group proteins, blood, plasma, serum of large mammals such as cows, pigs and horses, and These powders, egg whites or fractions / components thereof can be made into liquid preparations such as oral tablets, capsules, powders, granules, syrups, ointments, lotions and the like by using a usual formulation method.

When preparing a solid preparation for oral administration,
Excipients, binders, disintegrating agents, lubricants and the like are added to the native transferrin / lactoferrin group protein, and tablets, capsules, powders and granules are prepared by a conventional method.

When preparing a solution for oral administration, a buffer, a corrigent and a stabilizer are added to the main ingredient to prepare a liquid preparation by a conventional method.

When the chemotherapeutic agents for treating opportunistic infections are used in combination, they may be administered separately, or if necessary, they may be combined.

On the other hand, various foods such as milk, milk powder,
You may mix and administer in dairy products, such as yogurt and ice cream, and luxury items, such as chocolate and gum. or,
When administered to animals, it can be mixed with feed or pet foods. When administered to animals,
It can also be used as a raw material for feed and a premix material.

[0024]

[Example 1] A 32-year-old, 8-month-old man visited with fever, dyspnea, cough, cyanosis, etc. as a chief complaint, and an HIV-1 antibody was detected as a result of a blood test. A biopsy test of the lungs was performed on suspicion of Carini pneumonia. The diagnosis was confirmed because Pneumocystis carinii was detected in the lung biopsy specimen, and treatment was started immediately. A combination of trimethoprim and sulfamethoxazole was orally administered for 4 days, but the symptoms did not improve. Therefore, natural lactoferrin isolated from milk (purity about 9
(0%, iron saturation 26%) was orally administered once a day in an amount of 700 mg in combination with the above mixture before breakfast. The fever started to drop 48 hours after administration, and after 72 hours dyspnea, cough,
Cyanosis was also significantly improved. Along with this, the appetite also recovered, and on the 7th day after the administration of lactoferrin, the patient had a normal fever, and clinical symptoms such as dyspnea, cough and cyanosis disappeared. The patient was continuously administered lactoferrin, and the body weight recovered 3.5 kg in 4 weeks. During the administration, there were no side effects that were probably due to the administration of lactoferrin.

[0025]

Example 2 AIDS patients with recurrent stomatitis and gingivitis, 3 cases (male aged 35 to 42 years old) were treated with apolactoferrin (purity about 87%) separated and purified from milk. All patients had aftereffects or ulcers on the lips and gingival mucosa of the oral cavity, soft palate, or tongue,
The patient complained of anorexia due to severe pain, and tongue coating, stains and bad breath were observed. Before starting lactoferrin treatment, as a symptomatic treatment, silver nitrate solution application, adrenocortical hormone ointment application, and vitamin injection were performed, but there was almost no response to the treatment and it took a long time to recover. Granules containing 350 mg of apolactoferrin were orally taken once daily for 4 weeks before breakfast, and the progress was observed. In any case, inflammation was significantly improved from around the 6th day of administration,
Her appetite increased due to less bad breath and less pain. The ulcer and aphtha completely disappeared 8 to 14 days after the start of apolactoferrin administration, and the weight gain after 4 weeks was 3.1 kg (2.4, 3.6 and 3.) on average.
4 kg). In these cases, because the herpes simplex virus specific antigen and Candida albicans were found in the culture in the examination of the oral mucosa specimen,
Lactoferrin appears to have succeeded in eliminating these opportunistic pathogens from the affected area by improving the function of the biological defense system. Therefore, the therapeutic effect of lactoferrin on stomatitis and gingivitis due to AIDS immunodeficiency is clear. No side effects were observed.

[0026]

[Example 3] Carini pneumonia is the leading cause of death in AIDS patients, and it is said that approximately 30% of the patients die once they develop illness. Granules containing 700 mg of apo-type ovotransferrin extracted from egg white were orally administered once daily to the fasting fasting in the morning for 3 months to 3 adult male AIDS patients (23, 35 and 46 years old) who recovered from Carini pneumonia. During this time, the patient examined not only carinii pneumonia but also recurrence of infectious diseases caused by opportunistic pathogens such as cytomegalovirus, herpes simplex virus, and Candida albicans. As a result, Pneumocystis
No opportunistic infections, including carini, occurred and the quality of life of the patient was greatly improved. As a result of the patient being released from opportunistic infections,
Appetite increased, and body weight was 6.5 each compared to immediately before administration.
There was an increase of 4.0 and 10.8 kg. No side effects were observed.

[0027]

Example 4 The most common opportunistic infections in FIV-infected cats are chronic stomatitis, gingivitis, and then chronic upper respiratory tract disease. An attempt was made to treat a cat with chronic stomatitis and gingivitis who visited a veterinary hospital using a natural lactoferrin having a purity of about 85%. The daily dose of lactoferrin is 20 mg / kg.
The administration was performed in a divided manner by sprinkling on the affected area. There were various cases such as FIV-positive cases and gingivitis due to tartar. The administration period was at least 7 days, and the longest period was several months, and concomitant use of antibiotics was also performed as necessary. The effect was judged by improving clinical symptoms such as salivation and disappearance of bad breath. Of the 11 cases, 4 showed significant improvement in clinical symptoms, and 3 cases showed relief of symptoms only during the administration period (effective rate 64%). The remaining 4 cases are those with no effect or unknown cases. Two of the four cases in which clear improvement was observed were FIV-positive. In the FIV-positive cases, a wide range of severe gingivitis and stomatitis were observed in the oral cavity, and anorexia was exhibited due to pain, but relief of pain and disappearance of halitosis were observed from 7 to 10 days after administration. An increase in appetite was observed. When the administration of lactoferrin was stopped, salivation was observed again.

[0027]

Example 5 Each time an adult male cat with chronic upper respiratory tract disease caused by FIV infection came to the hospital, it was randomly divided into two groups, the first group was the ovotransferrin group, and the second group was the control group. . The experimental period was 8 weeks, during which the chemotherapeutic agent was freely administered. The dose of holo-type ovotransferrin was 20 mg / kg per day, which was given by mixing with canned pet food of fish meat or meat. The control cat died of pneumonia on days 8, 11 and 34 at the start of the experiment and only one survived. On the other hand, none of the ovotransferrin group died. Therefore, the therapeutic effect of chronic airway disease by ovotransferrin is clear, and the difference in mean survival time in both groups is statistically highly significant.

[0028]

EFFECTS OF THE INVENTION With the pharmaceutical composition provided by the present invention, treatment and prevention of opportunistic infections frequently occurring in immunodeficiency caused by lentivirus infection, which has been extremely difficult in the past, without fear of causing serious side effects. can do.

Claims (6)

[Claims] 1. A pharmaceutical composition for the treatment and prevention of opportunistic infections associated with lentivirus infection of an animal, which contains a mammalian transferrin-lactoferrin group protein in an unmodified form. 2. A pharmaceutical composition for the treatment and prevention of opportunistic infections associated with lentivirus infection, wherein the transferrin-lactoferrin group protein in claim 1 is lactoferrin obtained from mammalian milk. 3. A pharmaceutical composition for the treatment and prevention of opportunistic infections associated with lentivirus infection, wherein the transferrin-lactoferrin group protein in claim 1 is transferrin obtained from mammalian blood. 4. A pharmaceutical composition for the treatment and prevention of opportunistic infections associated with lentivirus infection, wherein the transferrin-lactoferrin group protein in claim 1 is ovotransferrin obtained from avian egg white. 5. A variety of transferrin-lactoferrin group proteins according to claims 1 to 4, which differ in iron saturation from a holotype completely saturated with trivalent iron ions to an apotype completely removed with iron ions. Is a transferrin-lactoferrin group protein, and is for the treatment and prevention of opportunistic infections associated with lentivirus infection. 6. The daily dose of the transferrin-lactoferrin group protein according to claim 1 is 0.1-100 mg / kg body weight, preferably 0.5-50 mg / kg body weight. A pharmaceutical composition for treating and preventing an opportunistic infection associated with a lentivirus infection.