WO2017202885A1

WO2017202885A1 - Treatment of viral infections by pulmonary delivery of oseltamivir carboxylate

Info

Publication number: WO2017202885A1
Application number: PCT/EP2017/062498
Authority: WO
Inventors: William COUET; Sandrine MARCHAND
Original assignee: INSERM (Institut National de la Santé et de la Recherche Médicale); Université de Poitiers; Centre Hospitalier Universitaire De Poitiers
Priority date: 2016-05-25
Filing date: 2017-05-24
Publication date: 2017-11-30

Abstract

The present invention relates to methods for the treatment of viral infections by pulmonary delivery of oseltamivir carboxylate. The inventors demonstrated in rats that much higher oseltamivir acid epithelial lining fluid than plasma concentrations may be obtained by direct nebulization of this active moiety, which may present a potentially major interest for the treatment of pulmonary infections due to influenza A and B viruses In particular, the present invention relates to a method of treating an infection caused by an orthomyxovirus in a subject in need thereof comprising administering by pulmonary delivery to the subject a therapeutically effective amount of oseltamivir carboxylate.

Description

TREATMENT OF VIRAL INFECTIONS BY PULMONARY DELIVERY

OF OSELTAMIVIR CARBOXYLATE

FIELD OF THE INVENTION:

The present invention relates to methods for the treatment of viral infections by pulmonary delivery of oseltamivir carboxylate.

BACKGROUND OF THE INVENTION:

Inhalation of antimicrobial agents may benefit to the treatment of lung infections by providing high concentrations at the site of infection and low systemic concentrations. However this potential advantage may vary with the type of nebulizer as well as with drug characteristics such as in particular membrane permeability. The inventors have recently developed a standardized experimental setting to compare the biopharmaceutical characteristics of nebulized antimicrobial agents in healthy rats, eventually leading to a biopharmaceutical classification of inhaled antimicrobial agents. The initial experiments have demonstrated major differences between antibiotics with high membrane permeability such as fluoroquinolones (1) and those with much lower membrane permeability, including colistin (2), tobramycin (3) and aztreonam (4) that are actually currently aerosolized in patients. But because pulmonary infections may also be due to viruses, the objective of the inventors was to compare the intrapulmonary pharmacokinetics of an anti-influenza drug after systemic administration and nebulization in order to assess the potential benefit of direct pulmonary delivery. Oseltamivir carboxylate (OC) is a potent and selective inhibitor of neuraminidase and is the most widely used antiviral drug for the treatment and prophylaxis of influenza A and B viruses (5). However OC presents a low membrane permeability, so in order to improve its oral bioavailability, it is administered peros in clinical practice as an ethyl ester prodrug, oseltamivir phosphate (OP). OP is rapidly absorbed due to its high membrane permeability and then rapidly hydrolyzed into the active OC (6). SUMMARY OF THE INVENTION:

The present invention relates to methods for the treatment of viral infections by direct pulmonary delivery of OC. In particular, the present invention is defined by the claims.

DETAILED DESCRIPTION OF THE INVENTION: The aim of this inventors was to determine the biopharmaceutical characteristics of OC after pulmonary delivery. After OC bolus and intra-tracheal nebulization in rats, blood was collected and bronchoalveolar lavages (BAL) were performed. Epithelial lining fluid (ELF) concentrations were estimated from BAL. ELF over plasma area under the curve (AUC) ratio was 842 times higher after NEB than after IV indicating that OC nebulization offers a biopharmaceutical advantage compared to IV administration.

Accordingly the first object of the present invention relates to a method of treating an infection caused by an orthomyxovirus in a subject in need thereof comprising administering by pulmonary delivery to the subject a therapeutically effective amount of oseltamivir carboxylate.

A virus of the Orthomyxoviridae family is encompassed by the term "orthomyxovirus". The virus of the Orthomyxoviridae family may be any genus of the six currently identified within the family, i.e. a virus of the genus Influenza A, Influenza B, Influenza C, Isavirus, Thogotovirus and/or a Quarandfil/Johnston Atoll/Lake Chad genus of virus or variants thereof. In some embodiments, the virus is an influenza virus of the Influenza A, Influenza B and/or Influenza C genera or sub-types thereof or variant forms thereof. A "variant form" of virus includes viruses having genomic elements derived from two or more sub-types of Influenza A, Influenza B and/or Influenza C or a sub-type thereof. Accordingly, the method of the present invention is particularly suitable for the treatment of an influenza infection. As used herein, the term "influenza infection" has its general meaning in the art and refers to the disease caused by an infection with an influenza virus. In some embodiments of the invention, influenza infection is associated with Influenza virus A or B. In some embodiments of the invention, influenza infection is associated with Influenza virus A. In some specific embodiments of the invention, influenza infection is cause by influenza virus A that is H1N1, H2N2, H3N2, H5N1, H7N7, H1N2, H9N2, H7N2, H7N3, and H10N7. Viral infection may be determined by measuring virus antibody titer in samples of a biological fluid, such as blood, using, e.g., enzyme immunoassay. Other suitable diagnostic methods include molecular based techniques, such as RT-PCR, direct hybrid capture assay, nucleic acid sequence based amplification, and the like. In some embodiments, a subject is a human infant. In some embodiments, a subject is a human child. In some embodiments, a subject is a human adult. In some embodiments, a subject is an elderly human. As used herein, the term "treatment" or "treat" refer to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse. The treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment. By "therapeutic regimen" is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy. A therapeutic regimen may include an induction regimen and a maintenance regimen. The phrase "induction regimen" or "induction period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease. The general goal of an induction regimen is to provide a high level of drug to a patient during the initial period of a treatment regimen. An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both. The phrase "maintenance regimen" or "maintenance period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a patient during treatment of an illness, e.g., to keep the patient in remission for long periods of time (months or years). A maintenance regimen may employ continuous therapy (e.g., administering a drug at a regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., disease manifestation, etc.]).

As used herein, the term "oseltamivir carboxylate" has its general meaning in the art and refers to (3R,4R,5S)-4-acetamido-5-amino-3-pentan-3-yloxycyclohexene-l-carboxylic acid also known as Ro64-0802 or GS4071 and which has the general formula of:

As used herein, the term "pulmonary delivery" has its general meaning in the art and refers to delivery of a drug to a subject by inhalation through the respiratory tract and into the lungs. In particular, pulmonary drug delivery may be achieved by nebulization. As used herein, the term "nebulization" refers to the generation of very fine liquid droplets for inhalation to the lungs by means of suitable devices called nebulizers.

Thus, in some embodiments, oseltamivir carboxylate is thus prepared as an aerosol composition. The term "aerosol composition" means a formulation that is suitable for pulmonary delivery. The aerosol composition is typically a suspension or slurry to be nebulized. The composition comprises pharmaceutical excipients. Excipients include but are not limited to (a) carbohydrates, e.g., monosaccharides such as fructose, galactose, glucose, D-mannose, sorbose, and the like; disaccharides, such as lactose, trehalose, cellobiose, and the like; cyclodextrins, such as 2-hydroxypropyl-.beta.-cyclodextrin; and polysaccharides, such as raffinose, maltodextrins, dextrans, and the like; (b) amino acids, such as glycine, arginine, aspartic acid, glutamic acid, cysteine, lysine, and the like; (c) organic salts prepared from organic acids and bases, such as sodium citrate, sodium ascorbate, magnesium gluconate, sodium gluconate, tromethamin hydrochloride, and the like; (d) peptides and proteins such as aspartame, human serum albumin, gelatin, and the like; and (e) alditols, such as mannitol, xylitol, and the like. A preferred group of carriers includes lactose, trehalose, raffinose, maltodextrins, glycine, sodium citrate, human serum albumin and mannitol.

By a "therapeutically effective amount" it is meant a sufficient amount to provide a therapeutic effect. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific polypeptide employed; and like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of the compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. However, the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day. Typically, the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated. A medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably from 1 mg to about 100 mg of the active ingredient. An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg/kg to about 20 mg/kg of body weight per day, especially from about 0.001 mg/kg to 7 mg/kg of body weight per day. A further object of the present invention relates to a system for the treatment of an infection caused by an orthomyxovirus, comprising a) a composition comprising oseltamivir carboxylate and b) an nebulizer wherein the composition is nebulized by the nebulizer to form an aerosol composition. As used herein, the term "nebulizer" has its general meaning in the art and refers to a device that creates aerosol which can be inhaled in response to the patient inhaling through a mouthpiece associated with a patient interface of the nebulizer. A variety of methods and devices are known in the art for nebulizing medication. Nebulizers produce a mist of drug- containing liquid droplets for inhalation. They are usually classified into two types: ultrasonic nebulizers and jet nebulizers. Single breath atomizers have also been developed (e.g., Respimat®), which is used to deliver a drug in a single inhalation and may be preferred because of less contamination. Jet nebulizers are more common and use a source of pressurized air to blast a stream of air through a drug-containing water reservoir, producing droplets in a complex process involving a viscosity-induced surface instability that leads to nonlinear phenomena in which surface tension and droplet breakup on baffles play a role. Ultrasonic nebulizers produce droplets by mechanical vibration of a plate or mesh. In some embodiments, the nebulizer comprises i) an aerosol generator comprising: a liquid storage container comprising the liquid pharmaceutical composition; a diaphragm having a first side and an opposite second side, the diaphragm having a plurality of openings extending therethrough from the first side to the second side, where the first side is connected to the liquid storage container such that the liquid filled into the liquid storage container comes into contact with the first side of the diaphragm; and a vibration generator capable of vibrating the diaphragm so that the liquid filled into the liquid storage container is atomized on the second side of the diaphragm through the openings of the diaphragm; ii) a mixing chamber into which the aerosol generator expels said aerosol, the mixing chamber in contact with the second side of the diaphragm; iii) an inhalation valve that is open to allow an inflow of ambient air into the mixing chamber during an inhalation phase and is closed to prevent escape of said aerosol from the mixing chamber during an exhalation phase; and iv) an exhalation valve that is open to allow the discharge of the respiratory air of a patient into the surroundings during the exhalation phase and is closed to prevent the inflow of ambient air during the inhalation phase.

The invention will be further illustrated by the following figures and examples. However, these examples and figures should not be interpreted in any way as limiting the scope of the present invention.

FIGURES: Figure 1. Predicted concentration-time profiles of oseltamivir carboxylate (6 mg.kg^"1) in plasma (solid line) and in ELF (dashed line) from simultaneous PK modeling after IV and NEB administrations. Closed and open symbols represent respectively mean + SD of experimental concentrations in plasma (total) and in ELF.

Figure 2. Concentrations of oseltamivir carboxylate in plasma ( ) and ELF (— ) after its administration by intravenous route (A) or nebulisation (B), and after administration of its oseltamivir phosphate prodrug by intravenous route (C) or nebulisation (D) in healthy rats, administered doses were equal to 6 mg/kg.

EXAMPLE 1: The animal experiments were conducted in compliance with EC Directive 2010/63/EU after approval by the local ethic committee (COMETHEA) and were registered by the French Ministry of Higher Education and Research under the authorization number 2015070211159865. The complete experimental setting was previously reported (1, 2). Briefly OC (Oseltamivir acid, Alsachim, Illkirch Graffenstaden, France) was administered under anesthesia at a dose close to 6mg.kg^_1 either by IV bolus into the tail vein (1 mL) or by intra-tracheal nebulization (NEB, 100 μί, microsprayer 1A-1B, Penn-Century, Wyndmoor, USA) in two groups of male Sprague Dawley rats (n = 17, 324 ± 23 g for IV group and n = 15, 322 + 20 g for NEB group, Janvier Laboratories, Le Genest-St-Isle, France). In both groups, broncho-alveolar lavage (BAL) and blood samples were collected 0.5, 2, 4 and 6 h after OC administration (3-5 rats per sampling time) and extra samples were collected at lh after IV administration. OC was assayed by LC-MS/MS using a previously described method with minor adaptation (7). A Shimadzu Nexera X2 separation module coupled with a ABSciex® API 3000 tandem mass spectrometer was used. Chromatographic separation was carried out on an X bridge BEH300 C18 300A column (5.0μπι, 150 x 2.1 mm ID, Waters, St- Quentin en Yvelines, France). The mobile phase was composed of acetonitrile/ammonium formiate lOmM/water (50: 10:40, v/v) and the flow rate was 0.2 ml.min^"1. The mass spectrometer was operated in the positive ion mode. Ions were analyzed by multiple reactions monitoring (MRM). Transition ions were m/z 285.1>138.0 for OC respectively and 289.2>138.0 (¹³C, ²H₃ OC) for deuterated internal standard. Ten-points calibration standards with concentrations between 0.625 and 375 ng.ml ¹ and 3 levels of control were prepared. The intra-day and inter-day variability in plasma and BAL was determined at three levels of concentrations with a precision and accuracy less than 15%. The limit of quantification (LOQ) in both media was set at 0.625 ng.ml^"1. Concentrations of urea in plasma and BAL were measured as previously described (2, 8). OC concentrations in epithelial lining fluid (CELF) were derived from measured BAL concentrations (CBAL) after correction by urea dilution(8). OC concentrations versus time in plasma and ELF were simultaneously analysed by a non-linear mixed effects method with S-ADAPT software (v 1.52). Two compartments were used to describe OC PK in plasma. OC PK in ELF was tested with one and two compartments, and two compartments with distinct volumes (VELFI, VELF₂) were kept in the final model as previously described with tobramycin (3). Compartments were connected by two directions equilibrium distribution clearances and by the addition of an efflux clearance from central compartment to ELF1 compartment (CLout). Only unbound drug in plasma was assumed to penetrate within ELF (4), but OC protein binding was considered to be negligible (6). Areas under plasma concentrations and ELF concentrations versus time-curves from 0 to infinity (AUCpiasma, AUCELF) were calculated from the model. Elimination half-lives (ti/2,_Piasma and ti/2,ELF) after IV administration and NEB were derived from the model (Berkeley Madonna, version 8.3.18, University of California).

This study was able to demonstrate a major effect of the route of administration on OC intrapulmonary PK, with much higher ELF concentrations after nebulization than after IV administration, and virtually identical plasma concentrations independently of the route of administration as illustrated on Fig 1. Lung exposure may be characterized by AUCELF estimated by the model ELF, which was 550-fold higher after nebulization than after IV administration. This effect of the route of administration may be further evidenced by the much higher ELF over plasma AUC ratio after nebulization ( AUCELF,NEB/ AUCpiasma,NEB = 842) than after IV administration (AUCELFJV / AUCpiasmajv = 1.01). Another interesting observation was that whatever the route of administration OC concentrations peaked early, more precisely at 0.5 h corresponding to the first sampling time, both in plasma and ELF. Elimination half-lives values in plasma were respectively equal to 1.8 h and 1.9 h after IV administration and nebulization, and to 2.3 h and 1.4 h in ELF (Fig. 1).

The important effect of the route of administration on OC pulmonary PK has already been observed with antibiotics such as colistin (2), tobramycin (3) and aztreonam (4), presenting low permeability (Pa_PP) across Calu-3 cells monolayers (9), but not with fluoroquinolones that are characterized by higher P_app (10). Therefore OC presents the characteristics of a low permeability drug, consistent with its poor oral availability which explains why it is not administered directly (11), and with its relatively low log D value at pH 7.4 (-2.1) (12). Unfortunately OC P_app across Calu-3 cells could not be properly estimated because of analytical limitations as previously encountered with tobramycin due to low permeability (3). This should be reassessed and complementary experiments should also be conducted after nebulization with OP in order to better appreciate the benefit of nebulizing OC directly instead of OP.

Although these results cannot be directly extrapolated to the clinical setting, they clearly demonstrate that much higher OC ELF than plasma concentrations may be obtained by direct nebulization of this active moiety, which may present a potentially major interest for the treatment of pulmonary infections due to influenza A and B viruses.

EXAMPLE 2: Initial experiments conducted after direct administration of oseltamivir carboxylate, the active moiety, showed that the ELF/plasma concentrations ratios was much higher after it's nebulisation (Fig 2,B) than after it's intravenous administration ( Fig 2, A), The new set of experiments has then shown that this ratio is also much higher after direct nebulisation of this active moiety (Fig 2,B) than after nebulisation of the prodrug (Fig 2,D). These new data also show that the route of administration is considerably reduced when oseltamivir phospahte is administered (Fig 2,C and Fig 2,D) instead of oseltamivir carboxylate (Fig 2,A and Fig 2,B), consistent with the differences in membrane permeability between the two compounds. Altogether these data confirm the anticipated biopharmaceutical advantage of nebulising directly the oseltamivir carboxylate, which thanks to it's reduced membrane permeability, allows attainment of much higher and sustained ELF concentrations than by any other type of administration.

REFERENCES:

Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.

1. Gontijo AV, Brillault J, Gregoire N, Lamarche I, Gobin P, Couet W, Marchand S. 2014. Biopharmaceutical Characterization of Nebulized Antimicrobial Agents in Rats. 1. Ciprofloxacin, Moxifloxacin and Grepafloxacin. Antimicrob Agents Chemother.

2. Gontijo AV, Gregoire N, Lamarche I, Gobin P, Couet W, Marchand S.

2014. Biopharmaceutical characterization of nebulized antimicrobial agents in rats: 2. Colistin. Antimicrob Agents Chemother 58:3950-3956.

3. Marchand S, Gregoire N, Brillault J, Lamarche I, Gobin P, Couet W.

2015. Biopharmaceutical Characterization of Nebulized Antimicrobial Agents in Rats: 3. Tobramycin. Antimicrob Agents Chemother 59:6646-6647.

4. Marchand S, Gregoire N, Brillault J, Lamarche I, Gobin P, Couet W.

2016. Biopharmaceutical Characterization of Nebulized Antimicrobial Agents in Rats. Antimicrob Agents Chemother.

5. Bardsley-EUiot A, Noble S. 1999. Oseltamivir. Drugs 58:851-860; discussion 861-852.

6. He G, Massarella J, Ward P. 1999. Clinical pharmacokinetics of the prodrug oseltamivir and its active metabolite Ro 64-0802. Clin Pharmacokinet 37:471-484. 7. Gupta D, Varghese Gupta S, Dahan A, Tsume Y, Hilfinger J, Lee KD, Amidon GL. 2013. Increasing oral absorption of polar neuraminidase inhibitors: a prodrug transporter approach applied to oseltamivir analogue. Molecular pharmaceutics 10:512-522.

8. Marchand S, Gobin P, Brillault J, Baptista S, Adier C, Olivier JC, Mimoz O, Couet W. 2010. Aerosol therapy with colistin methanesulfonate: a biopharmaceutical issue illustrated in rats. Antimicrob Agents Chemother 54:3702-3707.

9. Brillault J, De Castro WV, Harnois T, Kitzis A, Olivier JC, Couet W. 2009. P-glycoprotein-mediated transport of moxifloxacin in a Calu-3 lung epithelial cell model. Antimicrob Agents Chemother 53:1457-1462.

10. Brillault J, De Castro WV, Couet W. 2010. Relative contributions of active mediated transport and passive diffusion of fluoroquinolones with various lipophilicities in a Calu-3 lung epithelial cell model. Antimicrob Agents Chemother 54:543-545.

11. Samson M, Pizzorno A, Abed Y, Boivin G. 2013. Influenza virus resistance to neuraminidase inhibitors. Antiviral research 98: 174-185.

12. Hu ZY, Edginton AN, Laizure SC, Parker RB. 2014. Physiologically based pharmacokinetic modeling of impaired carboxylesterase- 1 activity: effects on oseltamivir disposition. Clin Pharmacokinet 53:825-836.

Claims

CLAIMS:

I. A method of treating an infection caused by an orthomyxovirus in a subject in need thereof comprising administering by pulmonary delivery to the subject a therapeutically effective amount of oseltamivir carboxylate.

2. The method of claim 1 for the treatment of an influenza infection.

3. The method of claim 2 wherein the influenza infection is associated with Influenza virus A or B.

4. The method of claim 2 wherein the influenza infection is caused by influenza virus A that is H1N1, H2N2, H3N2, H5N1, H7N7, H1N2, H9N2, H7N2, H7N3, and H10N7.

5. The method of claim 1 wherein the subject is a human infant.

6. The method of claim 1 wherein the subject is a human child.

7. The method of claim 1 wherein the subject is a human adult.

8. The method of claim 1 wherein the subject is an elderly human.

9. The method of claim 1 wherein the pulmonary drug delivery is achieved by nebulization.

10. The method of claim 1 wherein oseltamivir carboxylate is prepared as an aerosol composition.

I I. A system for the treatment of an infection caused by an orthomyxovirus, comprising a) a composition comprising oseltamivir carboxylate and b) an nebulizer wherein the composition is nebulized by the nebulizer to form an aerosol composition.

12. The system of claim 11 wherein the nebulizer comprises i) an aerosol generator comprising: a liquid storage container comprising the liquid pharmaceutical composition; a diaphragm having a first side and an opposite second side, the diaphragm having a plurality of openings extending there through from the first side to the second side, where the first side is connected to the liquid storage container such that the liquid filled into the liquid storage container comes into contact with the first side of the diaphragm; and a vibration generator capable of vibrating the diaphragm so that the liquid filled into the liquid storage container is atomized on the second side of the diaphragm through the openings of the diaphragm; ii) a mixing chamber into which the aerosol generator expels said aerosol, the mixing chamber in contact with the second side of the diaphragm; iii) an inhalation valve that is open to allow an inflow of ambient air into the mixing chamber during an inhalation phase and is closed to prevent escape of said aerosol from the mixing chamber during an exhalation phase; and iv) an exhalation valve that is open to allow the discharge of the respiratory air of a patient into the surroundings during the exhalation phase and is closed to prevent the inflow of ambient air during the inhalation phase.