METHOD FOR ORAL TRANSMUCOSAL DELIVERY OF
INTERFERON
FIELD OF THE INVENTION The present invention relates to a method for oral transmucosal delivery of interferon.
BACKGROUND OF THE INVENTION
Interferons (IFNs) are naturally occurring proteins with antiviral, antiproliferative and immunoregulatory activity. The following definition for interferon has been accepted by international committee assembled to devise a system for the orderly nomenclature of interferons: "To qualify as an interferon a factor must be a protein which exerts virus nonspecific, antiviral activity at least in homologous cells through cellular metabolic processes involving synthesis of both RNA and protein." J. Interferon Research. 1: pp. vi (1980). Four distinct classes of interferons are known to exist in humans. Pestka et al., Ann. Rev. Biochem.. 56: 727 (1987): Emanuel and Pestka, J. Biol. Chem. 268: 12565 (1993). The three main human interferons are known as IFN-alpha, IFN-beta and IFN-gamma. The IFN-alpha family represents the predominant class of human IFNs. Intramuscular administration is a predominant route of interferon delivery to a systemic circulation of mammals.
Delivery of drugs through the oral absorptive mucosa is a very attractive and feasible site for systemic delivery of interferons. The major advantages of this route for systemic action of a drug formulation are that it bypass the hepatic first-pass metabolism and avoids pre-systemic elimination in the gastrointestinal tract. Furthermore, the oral cavity is a highly acceptable by patients.
Surprisingly, we have now found that administering interferon into a mammal's oral cavity under tongue in aerosol form manifests much more pronounced raising the blood level of interferon than the same dose of interferon in a non-aerosol form and under otherwise equal conditions.
It is an object of the present invention to provide the improved method for oral transmucosal delivery of interferon.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a method for oral transmucosal delivery of interferon comprising the steps of: (a) providing an interferon formulation comprising a therapeutically effective amount of interferon, (b) administering said interferon formulation in form of aerosol consisting of solid particles or liquid droplets with mass median aerodynamic diameter from 4 to 150 μm sublingually into a mammal's oral cavity, and (c) delivering said interferon by absorption through a mammal's oral mucosal tissue.
A particular advantage of the present invention is that the level of interferon in systemic circulation achieved in a mammal with an amount of interferon when administered into a mammal's oral cavity in form of aerosol is greater than the level achievable with the same amount of interferon administered into a mammal's oral cavity in non-aerosol form and under otherwise equal conditions. Accordingly, it is now possible through the practice of this invention to achieve certain desired therapeutic effects using less amount of interferon than was heretofore possible. The desired therapeutic effects achievable through the practice of this invention include all known in the art therapeutic effects of interferon. Such effects include, but are not limited to, antiviral, antiproliferative, antitumor, antibacterial, and immunoregulatory action of interferon in mammals. Therefore, in practicing this invention, it is possible to minimize potential adverse effects, which may be associated with larger, therapeutic doses of the interferon and still achieve the therapeutic effect.
As used herein, the term "aerosol" means a colloidal system consisting of very finely divided solid particles or liquid droplets dispersed in and surrounded by a gas.
The present invention is not limited in any way to a specific device for aerosol preparation but is applicable to all such devices now known or subsequently developed. The therapeutically effective amount of interferon is within the skill of those who practice in the art having the benefit of the disclosure herein. Typically, interferon will be present in method of the invention in amounts within
its normal or less dosage unit and daily regimen ranges as detailed in medical literature. The dosage range will be from about 100 IU to about 5x106 IU interferon per mammal. Preferably, mammals are administered about 3x106 IU human recombinant interferon-alpha per mammal per day. The present invention is not limited in any way to specific interferon but is applicable to all such interferon now known or subsequently discovered or developed. Nonetheless, a preferred interferon for use in the method of this invention is human recombinant interferon-alpha.
The interferon formulation may be formulated with various pharmaceutically acceptable carriers or diluents in the form of liquid aerosols, solid aerosols, or sprays. Such carriers can include solid diluents, sterile aqueous media, non-toxic organic solvents, and etc.
The interferon formulation may be formulated with various pharmaceutical ingredients known from the art. These ingredients may include, but are not limited to, buffering agents (such as phosphate buffer, carbonate buffer, tris buffer, tartrate buffer, borate buffer, acetate buffer, or maleate buffer), colorants, flavorants, coating agents, suspending agents, sweetening agents, anti-fungal preservatives, antimicrobial preservatives, clarifying agents, antioxidants, surfactants, and tonicity agents. The interferon formulation may be formulated with various pharmaceutically acceptable absorption promoters known from the art. Such absorption promoters may include, but are not limited to, as lecithin, fatty acids, chitosan, salicylates, amino acids, emulsifiers like polyoxyethylene alkyl esters and sodium lauryl sulfate, caproic acid, lactic acid, malic acid, citric acid, pyrrolidonecarboxylic acid, N-alkylpyrrolidones, benzalkonium chloride, cetylpyridinium chloride, cetyltrimethylammonium bromide, cyclodextrins, dextran sulfate, lauric acid, lysophosphatidylcholine, menthol, methoxysalicylate, methyl oleate, polyoxyethylene, polysorbate, sodium glycocholate, sodium glycodeoxycholate, sodium taurocholate, sulfoxides, various alkyl glycosides, etc. The following examples are presented to demonstrate the invention. The examples are illustrative only and are not intended to limit the scope of the invention in any way.
EXAMPLE 1
This example shows that sublingual administration of interferon in form of aerosol resulted in improved transmucosal delivery of interferon.
Materials. Human recombinant interferon-alpha2b (rhlFN-alpha) was from (Cytokine Ltd., St. Petersburg, Russia).
Assay. Human interferon-alpha ELISA assay (Cytokine Ltd., St. Petersburg, Russia) was used. Mice. Male C57BL mice 10- to 12-week aged were used.
Procedure. Mice were subdivided into three groups. Mice in the first group (control 1) received sublingually solution of 2.5 μg (0.5χ106 IU) rhlFN-alpha dissolved in 0.1 ml of saline (pH 7.2). Mice in the second group (control 2) received intramuscularly 2.5 μg (0.5x106 IU) of rhlFN-alpha dissolved in 0.1 ml of saline (pH 7.2). Mice in the third group received sublingually 2.5 μg (0.5x106 IU) rhlFN-alpha in form of liquid aerosol by the following method: parent solution of 2.5 μg (0.5x106 IU) rhlFN-alpha was prepared by dissolution of 2.5 μg (0.5x106 IU) rhlFN-alpha in 0.1 ml of saline (pH 7.2), this solution was sprayed by nebulizer to form liquid aerosol consisting of liquid droplets with mass median aerodynamic diameter from 8 to 150 μm, and this aerosol was administered to field under tongue in mouse's oral cavity. Absorption of rhlFN-alpha through oral mucosa into systemic circulation was assayed in the course of time with human interferon alpha ELISA.
Data are presented in Table 1 as mean ± SD (n = 5) of rhlFN-alpha levels in plasma of mice in comparison with control 1 and control 2.
Table 1. Time course of the plasma rhlFN-alpha levels.
*Differs significantly of Control 1 (p < 0.001) 'Differs significantly of Control 2 (p < 0.001)
As shown in Table 1 , the sublingual administration of interferon in liquid aerosol form resulted in improved transmucosal delivery of interferon into systemic circulation which delivery is significantly much greater as compared to administration of the same amount of interferon in liquid sublingual non-aerosol form and under otherwise equal conditions (control 1).
Furthermore, the sublingual administration of interferon in liquid aerosol form resulted in improved transmucosal delivery of interferon into systemic circulation which delivery is significantly much faster and much greater as compared to intramuscular administration of the same amount of interferon and under otherwise equal conditions (control 2).
EXAMPLE 2
This example shows that sublingual administration of interferon formulation comprising interferon and an absorption promoter in form of aerosol resulted in improved transmucosal delivery of interferon.
Procedure. Mice were subdivided into two groups. Mice in the first group (control) received sublingually solution of 2.5 μg (0.5χl06 IU) rhlFN-alpha dissolved in 0.1 ml of saline (pH 7.2) with addition of 0.01 mg soy lecithin as the absorption promoter. Mice in the second group received sublingually interferon formulation comprising 2.5 μg (0.5x106 IU) rhlFN-alpha in form of liquid aerosol by the following method: parent solution of 2.5 μg (0.5x106 IU) rhlFN-alpha was prepared by dissolution of 2.5 μg (0.5x106 IU) rhlFN-alpha in 0.1 ml of saline (pH 7.2) with addition of 0.01 mg soy lecithin as the absorption promoter, this solution was sprayed by nebulizer to form liquid aerosol consisting of liquid droplets with mass median aerodynamic diameter from 8 to 150 μm, and this
aerosol was administered to field under tongue in mouse's oral cavity. Absorption of rhlFN-alpha through oral mucosa into systemic circulation was assayed in the course of time with human interferon alpha ELISA.
Data are presented in Table 2 as mean ± SD (n = 5) of rhlFN-alpha levels in plasma of mice in comparison with control.
Table 2. Time course of the plasma rhlFN-alpha levels.
*Differs significantly of Control (p < 0.001)
As shown in Table 2, the sublingual administration of interferon formulation comprising interferon and an absorption promoter in liquid aerosol form resulted in improved transmucosal delivery of interferon into systemic circulation which delivery is significantly much greater as compared to administration of the same amounts of interferon and the absorption promoter in liquid sublingual non-aerosol form and under otherwise equal conditions (control).