MXPA97006098A - Product and method for preventing respiratory viral infection - Google Patents

Abstract

The present invention relates to a method for treating pneumonia or other respiratory disease caused by respiratory syncytial virus, adenovirus, parainfluenza virus or influenza virus by orally administering a liquid, preferably a infant formula, containing an antibody to neutralize the vir

Description

PRODUCT AND METHOD FOR PREVENTING RESPIRATORY VIRAL INFECTION BACKGROUND OF THE INVENTION FIELD OF THE INVENTION This invention relates to an orally administered antibody with neutralizing activity of respiratory syncytial virus (RSV), adenovirus, parainfluenza virus or influenza virus and its use for decrease the incidence or severity of RSV or other viral infections of the upper and lower respiratory tract. Description of the Prior Art Respiratory syncytial virus is the main cause of pneumonia and bronchiolitis in childhood. Infants between the ages of two and five months have the most severe disease and may require hospitalization. More than half of all infants become infected with RSV during their first year of exposure and almost all are infected after a second year. Children who went to day care centers tend to have more severe infections and at an earlier age. Infections with repeated RSV are common, although repeated episodes tend to be less severe. During seasonal epidemics, most infants, children and adults are at risk of infection or reinfection. In addition to infections in healthy infants and children, other groups at risk for serious RSV infections include premature infants, hospitalized children, infants and children. children with heart or lung disorders, immunocompromised children and adults, and the elderly. The symptoms of RSV infection vary from moderate to severe bronchiolitis and pneumonia. Respiratory syncytial virus has also been associated with acute otitis media and RSV can be recovered from middle ear fluid. Respiratory syncytial virus is an RNA virus that can produce cellular (syncytial) fusion in tissue culture. It is classified as a pneumovirus within the paramyxovirus family. The RNA genome encodes at least 10 proteins including two matrix proteins in the viral envelope (Ryan, Sherris' Medical Microbiology, 3rd ed., Appleton and Lange, p.458, 1994). A matrix protein forms the inner lining of the viral envelope. Antigens on the surface of the envelope are glycoprotein G, the likely binding site to the host cell receptors and the F-glycoprotein that induces fusion. The glycoprotein G antibodies can neutralize the virus in vitro. Infection with the virus causes humoral IgG and IgA responses and secretory antibody. Immunity is not permanent and repeated infections are common, however, the severity of the disease tends to decrease with increasing age and with successive reinfection. No vaccine has been shown to protect against RSV and antiviral drugs so far have only limited utility. Maternal breastfeeding may offer some protection against RSV infection. The specific IgA and IgG antibodies to RSV have been found in human milk and colostrum and RSV neutralization in cell cultures can be achieved with both immunoglobulin components with or without human milk immunoglobulin (Laegreid et al., "Neutralizing Activity in Human Milk Fractions against Respiratory Syncitial virus, Acedia Paediatrica Scandinavica, 75-696-701, 1986.) Okamato et al., (Acedia Paediatrica Scandinavica Supplement, 351: 137-143, 1989) report that immunity acquired by an infant Whether it is through the placenta or through breastfeeding, it can reduce the risk of lower respiratory tract disease.The focus of the report by Okamato and others, is on the role of maternal antibodies transmitted in breast milk and the possible role of breast milk to modulate an immune response of the infant to RSV The approach of the present invention is to a method for producing additional passive immunity neutralizing antibodies to a product that will be orally ingested. Ingested orally, as used herein, refers to a substance that is ingested by the host. Previous treatments for RSV infection have depended on both parenteral and aerosol administration of agents such as monoclonal antibodies or virucidal drugs such as ribavirin. The present invention describes oral administration of an antibody with RSV neutralizing activity.

Prince et al. (U.S. Patent No. 4,800,078) teaches a method for topical application of antibodies to RSV in the lower respiratory tract, preferably by administering immunoglobulins as a small particle aerosol. Immunoglobulins can also be administered by the intravenous route. In U.S. Patent No. 5,290,540, Prince et al. Describe the topical administration in aerosol form into small particles of both an anti-inflammatory agent and an anti-infective agent in the pneumonia treatment caused by bacetaps or viruses including RS CA 2,040,770 to Young et al., describes a process for the treatment of respiratory viruses, including RSV, by administering a non-neutralizing monoclonal antibody against a fusion protein of RSV (the ghcoprotein F). of monoclonal antibody using the method of Young and others can be topical and administered intranasally or by aerosol breathing, or systemic by intramuscular administration. In contrast, the present invention describes the orally administered treatment WO 92/01473 describes the treatment of viral disease of the lower respiratory tract using the small particle aerosol method to deliver neutralizing and / or therapeutic monoclonal antibodies to specific viral surface antigenic sites WO 92/19244 teaches the combination of an anti-infective agent such as immunoglobulin G from humans or an antibiotic combined with an anti-inflammatory agent or corticosteroids delivered to the respiratory tract in the form of small particle aerosol. WO 94/17105 discloses chimeric human-murine antibodies with high neutralizing activity specific against RSV, preferably against the F antigen of RSV. References in the prior art describe the delivery of a virus neutralizing compound either topically by inhalation of a small particle aerosol or parenterally by intravenous and intramuscular injection. The feeding of a non-absorbed RSV neutralizing compound to avoid or decrease the incidence and severity of RSV infection has not been described or demonstrated in prior art references. This concept, as demonstrated in the present invention, depends on the ability of a neutralizing antibody to RSV to decrease the viral load on mucosal surfaces of the nasopharynx, oropharynx and hypopharyngeal and, therefore, prevents or decreases the diffusion of infectious virus from nose to lungs when the antibody is ingested. The delivery of a neutralizing antibody to RSV in a liquid product is particularly advantageous because of the ease of administration. DESCRIPTION OF THE INVENTION The invention is an orally administered liquid product that contains a respiratory virus neutralizing compound. In one embodiment of the invention, the virus neutralizing compound is a neutralizing antibody to RSV, which is added to a nutritional product. The invention is also a method for delivering an effective concentration of the respiratory virus neutralizing antibody by adding it to a liquid product. used herein and in the claims, a neutralizing antibody is meant to mean an antibody from any mammalian source such as human or bovine that can neutralize the respiratory virus. In one embodiment of the invention, the respiratory virus neutralizing antibody is added. to a infant product, such as infant formula, and the infant is fed during the first year of life. The infant formula can be a powder for reconstitution with water, a ready-to-feed liquid or a concentrated liquid. The respiratory viruses to which can apply the invention, include respiratory syncytial virus atorio, adenovirus, parainfluenza virus and influenza virus PROTOCOL EXPERIMENTAL Studies will be carried out to determine the impact of Feeding neutralizing antibodies against lung infection induced by human respiratory syncytial virus (RSVH) in animals The objectives of the studies include the identification of an animal model for nasal confrontation with RSVH in order to evaluate the influence of dietary feeding on pulmonary infection of RSVH and the determination if the feeding of neutralizing antibodies to RSVH and / or other neutralizing compounds can prevent or mitigate the lung infection in the nasal confrontation with RSVH. A positive result with an animal model will eventually allow the clinical evaluation of an increased liquid product with a neutralizing compound. of VSRH. Animals and diet: Cotton rats of the same 30-day-old parents (Sigmodon fulviventer) free of serum neutralizing antibody against human respiratory syncytial virus (RSVH) will be used. The rats will be fed a liquid basal diet consisting of infant formula for two days before the experiments begin. One day before the intranasal inoculation of viruses experimental diets will be provided. The consumption of food and change of body weight will be monitored daily. When the study is finished, all rats will be killed by asphyxiation with carbon dioxide and the nasal and lung tissue will be removed for analysis. Virus: The virus that will be used is human respiratory syncytial virus of subgroup A2 (VSRH / Long). The virus will be prepared by infecting monolayers of HEp-2 cells, which will be developed until the monolayers show approximately 9% syncytium formation. The medium of the monolayers will be collected, combined and clarified by centrifugation at 450 x g. The clarified supernatant fluid will be passed through a 0.45 μM filter. This supernatant will contain human respiratory syncytial virus (RSV) at 10 6 PFU / ml as determined by plaque analysis. Antibodies: The commercially obtained RSVH antibodies (VSRHI) (Sandoz, East Hanover, NJ) will be incorporated in liquid diets at various concentrations and the in vitro neutralizing activity of the experimental diets supplemented with RSVRSV) will be determined by plaque reduction assays. Virus titration: The oropharyngeal swab will be taken daily before 8 a.m. from the second day of virus inoculation until the end of the experiment. The VSRH antigen will be determined in all the swabs. At necropsy, nasal and lung tissue will be homogenized in 10 parts (weight / volume) of balanced Hanks salt solution supplemented with 0.218 M sucrose, 4.4 mM glutamate, 3.8 mM KH2P04, 3.2 mM K2HP04. The resulting suspension will be used to determine virus titers by plaque analysis in monolayers of Hep-2 cells. Histopathological examination: Nasal tissues fixed with formalin and lungs, will be embedded in paraffin, cut into coronal sections, stained with hematoxylin-eosin with periodic acid-Schiff (APS), and examined under a light microscope. The slides will be prepared by a pathologist for whom the sample numbers will be capped during the microscopic examination. The histopathology of the lung stained by APS will be classified from 0-2.0, 2.1-6.0, 6.1-10.00, 10.1-12 as defined by Piedra et al. ("Mechanism of lung damage in cotton rats immunized with respiratory syncytial virus inactivated in formalin ", Vaccine 7: 34-38, 1989).

Statistical analysis single-tailed X will be used to compare proportions of measurements between independent groups Vapanza analyzes will be used to compare virus titers and severity of damage to the lung. Example The study to examine the dose-response relationship between the immunoglobulin of the human respiratory syncytial virus (VSRHIG) of the diet and infection of human respiratory syncytial virus (RSVH) Piazza and others ("Immunotherapy of respiratory smcytial virus infection in cotton rats (Sigmodon fulviventer) using IgG in a small-particle aerosol", Journal of Infectious Disease 166 1422-1424, 1992) have shown that VSRHIG in a 5mg / 100ml solution administered for 15 minutes in a small particle aerosol after intranasal inoculation of cotton rats with RSVH reduced their virus titration 50 times The present study is designed to determine if the oral administration of VSRHIG can reduce similarly This will be done by incorporating an anti-RSV IgG in a liquid diet at various concentrations, after which the m vitro neutralizing activity of the experimental diets will be determined with the plaque reduction assay as described by Ppnce and others. ('The pathogenesis of respiratory syncytial virus infection in cotton rats' American Journal of Pathology 93 771-792 1978) For the study 70 rats will be divided into 7 treatment groups Treatment group 1 will comprise 10 rats and serve as a negative control. They will be fed with the basal liquid diet and inoculated with 0.1 ml of supernatant of the culture medium of Hep-2 cells that lack cells or VSRH. Ten rats in each of the treatment groups 2, 3, 4 or 5, will be fed with the basal liquid diet supplemented with VSRHIG at 0, 0.5, 5 and 10 mg / 100 ml respectively. Assuming that each rat will consume 100 ml of liquid diet, the anticipated daily dose for rats in each group should be 0, 0.5, 5, and 10 mg of RSVRS. Rats in treatment groups 6 and 7 will be fed the liquid diet supplemented with other agents that will be tested for antirespiratory virus properties. One day after consuming the experimental diets, the rats in treatment groups 2-7 will be inoculated intranasally with RSV / Long using 0.1 ml of viral suspension containing RSV at 103 PFU / ml (plaque forming units). All rats will continue to consume their assigned diets until the study ends. Four days after being confronted with the virus, all rats will be killed and the nasal tissues and lungs will be removed for sequential analysis. An effective concentration of the respiratory virus neutralizing antibody can be added to a liquid product. In a specific embodiment of the invention, the liquid product is a infant formula that can be fed to an infant during the first year of life, the period when the infant is more vulnerable to RSV infection. Children's nut formulations could be in the form of powder for reconstitution with water, a liquid ready to be fed, or concentrated liquid. However, it should be understood that the scope of the present invention is not limited to these specific modalities. The invention can be practiced in another way than is particularly described and still be within the scope of the appended claims.

Claims (8)

1. CLAIMS 1. A method to prevent or treat upper respiratory and / or lower respiratory tract infections caused by a virus selected from the group consisting of respiratory syncytial virus, adenovirus, parainfluenza virus and influenza virus, which method comprises: a . adding a therapeutically effective amount of an antibody with respiratory virus neutralizing activity to a liquid; and b. orally administering the liquid to a mammal.
2. 2. A method for preventing or treating respiratory virus infections according to claim 1, wherein the antibody is IgG.
3. 3. A method for preventing or treating respiratory virus infections according to claim 1, wherein the antibody is IgA.
4. 4. A method for preventing or treating respiratory virus infections according to claim 1, wherein the liquid is infant formula.
5. 5. A method for preventing or treating respiratory virus infections according to claim 1, wherein the mammal is an infant.
6. 6. A method to prevent or treat infections by Respiratory Syncytial Virus of the upper and / or lower respiratory tract whose method comprises a. adding a therapeutically effective amount of an antibody with neutralizing activity of Respiratory Syncytial Virus to a liquid; and b. orally administering the liquid to a mammal.
7. 7. A liquid for the prevention or treatment of respiratory virus infection in a mammal comprising a therapeutically effective amount of a respiratory virus neutralizing antibody.
8. 8. A infant formula comprising a therapeutically effective amount of a respiratory virus neutralizing antibody.