MXPA97009866A - Method for the treatment of pulmon diseases using oligonucleotides antisent - Google Patents

Abstract

A method for treating respiratory tract disease in a subject in need of such treatment is described. The method comprises topically administering to the subject an antisense oligonucleotide in an amount effective to treat airway disease, wherein the antisense oligonucleotide is essentially free of adenosine. Pharmaceutical formulations are also described

Description

METHOD FOR THE TREATMENT OF LUNG DISEASES USING ANTI-AGING OLIGONUCLEOTIDES This invention was made with the help of the Government under RO1CA47217-06 of the National Cancer Institute. The Government has certain rights for this invention.

FIELD OF THE INVENTION This application relates to a method for administering antisense oligonucleotides essentially free of adenosine as a treatment for lung diseases.

BACKGROUND OF THE INVENTION Antisense oligonucleotides have received considerable theoretical consideration as potentially useful pharmacological agents in human diseases. R. Wagner, Wature 372, 333-335 (1994). However, the practical applications of these molecules and current models of human disease have been difficult to find. An important consideration in the pharmacological application of these molecules is the route of administration. Most experiments using antisense oligonucleotides in vivo have involved direct application to limited brain regions (see C. Wahlestedt, Trends in Pharmacology cal Sciences 15, 42-46 (1994); J. Lai et al., Neuroreport 5 , 1049-1052 (1994), K. Standifer et al., Neuron 12, 805-810 (1994) '; A. Akabayashi et al., Brain Research 21, 55-61 (1994)), or the spinal fluid ( see, for example, L. Tseng et al., European J. Pharmacol 258, Rl-3 (1994), R. Raffa et al., European J. Pharmacol. 258, R5-7 (1994), F. Gillardon et al. al., European J. Neurosci 6, 880-884 (1994)). Such applications have limited clinical utility due to their invasive nature. The systematic administration of antisense oligonucleotides also has significant problems with respect to pharmacological application, the greatest of which is the difficulty to mark tissues involved in the disease. In contrast, the lung is an excellent potential target for the application of antisense oligonucleotides since this can be approximated non-invasively and in a tissue-specific manner. Additionally, the lung represents an exceptional target for antisense ODN therapeutics compared to other target organs or tissues in vivo, possibly since the lung is serialized with surfactant which consists mainly of cationic lipids, well known to improve cellular uptake of ODN in other systems. However, the technology involved in the delivery of antisense agents to the lung remains relatively undeveloped, and potential problems related to the application of antisense agents remain unexplored. Adenosine, a purine which contributes to intermediary metabolism and participates in the regulation of physiological activity, is a recognized neuromodulator. This nucleoside is involved in many local regulatory mechanisms, particularly in synapses in the CNS and in the neuroeffector junctions in the periphery. In the CNS, adenosine is known to inhibit the release of a variety of neurotransmitters (noradrenaline, serotonin, GABA, acetylcholine, dopamine, glutamate, etc.), to inhibit neurotransmission, decrease neuronal depression, induce spinal analgesia and possess properties. Anxiolytics (ES Ben- Soreket al.'V Archives of Internal Medicine 153, 2701-2702 (1993)). In the heart, adenosine is known for slow atrioventricular conduction (AV), pacemaker suppressor activity, has antiarrhythmic effects, modulates autonomic control, and triggers the synthesis and release of prostaglandins. M.K. Church et al., J. Allergy & Clinical Immunology 92, 190-194 (1993). This also has potent vasodilator effects and modulates vascular tone. S.T. Holgate et al., Annals of the New York Academy of Sciences 629, 227-236 (1991).

As a therapeutic agent, adenosine has achieved considerable recent success as an antiarrhythmic agent in the treatment of supraventricular tachycardia. See C.G. DeGroff and M.J. Silka, Journal of Pediatrics 125, 822-823 (1994); I. Drake et al., Human and Exp. Toxicol 13, 263-265 (1994). However, many adverse effects of the treatment of adenosine have been reported in the literature. See, for example, A. Aggarwal, et al., Anesthesiology 79, 1132-1135 (1993); K.K. Burkhart, American J. Emergency Med. 11, 249-250 (1993); S.K. Srinivasan and P.J. Iversen, J. Cl in. Lab. Analysis 9, 129-137 (1995); AC Stein et al., Pharmacology & Therapeutics 52, 365-384 (1991); B.B. Fredholm et al., Phar Acological Reviews 46, 143-156 (1994); H. Saito, et al., Blood 66, 1233-1240 (1985). In particular, asthmatic individuals show extreme sensitivity to adenosine and adenosine monophosphate. See, J.H. Butterfield et al., Leukemia Res. 12, 345-355 (1988); CLONETICS: Normal Human Cell Systems Manual (1995); R.W. Wagner, Nature 372, 333-335 (1994). Serious, near-fatal induction of bronchospasm has occurred in asthmatic individuals administered with adenosine for supraventricular tachycardia. See, S. Tabor, in: Current Protocols in Molecular Biology, Vol. 1, Section 3.10.2 (John Wiley &Sons, 1987); J.H. Weiss, Id., In Section 6.2.2.

Similarly, asthmatic rabbits produced using rabbit model dust allergic to human asthma akars also show that they respond to adenosine sprayed with marked bronchoconstriction, whereas non-asthmatic rabbits show no response. S. Ali et al., Agen ts Actions 37, 165-176 (1992). Recent work using this model system has suggested that bronchoconstriction and bronchial hyperresponsiveness induced by adenosine in asthma are mediated primarily through the stimulation of adenosine receptors. S. Ali et al., J. Pharmacol. Exp. Ther 268, 1328-1334 (1994), S. Ali et al., Am. J. Physiol 266, L271-277 (1994). Adenosine is contraindicated in the lungs of asthmatics (who represent 10% of adults and 5.5% of the pediatric population in the United States.) Since antisense ODNs are typically composed of four base pairs, adenine, guanine, cytosine, and thymidine , its decomposition products will produce free deoxyadenosine monophosphate in these hypersensitive airways Deoxyadenosine monophosphate differs from adenosine monophosphate only by the loss of an oxygen atom on the 3 'carbon of the sugar portion.

BRIEF DESCRIPTION OF THE INVENTION A first aspect of the present invention is a method for treating respiratory tract disease in a subject in need of such treatment. The method comprises administering an antisense oligonucleotide substantially free of adenosine to the lungs of the subject in an amount effective to treat the disease of the respiratory tract. A second aspect of the present invention is a pharmaceutical composition, comprising, together with a pharmaceutically acceptable carrier, an antisense oligonucleotide essentially free of adenosine in an amount effective to treat a disease of the respiratory tract. A third aspect of the present invention is the use of an antisense oligonucleotide essentially free of adenosine as set forth above for the preparation of a medicament for treating respiratory tract disease in a subject in need of such treatment.

BRIEF DESCRIPTION OF THE DRAWINGS Figures 1-4 demonstrate that antisense oligonucleotides can be used as effective agents in the treatment or prevention of diseases of the respiratory tract. Figure 1 illustrates the effects of antisense oligonucleotides on the A-adenosine receptor and antisense oligonucleotides for the control of mismatches on the dynamic adaptability of the bronchial airways in a rabbit model. Figure 2 illustrates the specificity of the antisense oligonucleotides at the A-adenosine receptor A by the number of adenosine A- and A2-receptor present in airway tissue treated with antisense oligonucleotides at the adenosine receptor.

Al- Figure 3 is a graphic representation illustrating that deoxyadenosine monophosphate sprayed is a potent bronchodilator in the asthmatic airways of allergic rabbits. In addition, the figure shows that the effect of deoxyadenosine monophosphate is equipotent to that observed for adenosine monophosphate. Figure 4 is a graphical representation illustrating that bronchoconstrictor effects occur with sprayed phosphorothioate oligodeoxynucleotides containing adenosine, but not oligodeoxynucleotides that are free of adenosine.

DETAILED DESCRIPTION OF THE INVENTION The nucleotide sequences are presented here only of one strand, in the 5 'to 3' direction from left to right. Nucleotides and amino acids are represented herein as recommended by the IUPAC-IUB Biochemical No enclature Commission, or (by amino acids) by the three-letter code, in accordance with 37 CFR §1.822 and established use. See, for example, Pa tentln User Manual, 99-102 (November 1990) (United States Patent and Trademark Office, Office of the Assistant Patent Commissioner, Washington, D.C. 20231); U.S. Patent No. 4,871,670 to Hudson et al. in Column 3-lines 20-43 (the applicants specifically propose that the description of this and all other reference patents cited herein be incorporated herein by reference). The method of the present invention can be used to treat airway disease in a subject for any reason, with the intention that the adenosine content of antisense compounds be eliminated or reduced to prevent its release after antisense degradation. Such release can cause serious bronchoconstriction, including life-threatening in patients with hyperreactive airways. Examples of respiratory diseases that can be treated by the method of the present invention include cystic fibrosis, asthma, chronic obstructive pulmonary disease, bronchitis, and other respiratory diseases characterized by an inflammatory response. The antisense oligonucleotides for the A- ^ and A3 receptors are shown to be effective in down-regulation of A- ^ or A3 in the cell. A novel aspect of this treatment, compared with traditional treatments for adenosine-induced bronchoconstriction, is that administration is direct to the lungs. Additionally, a protein by itself receptor is reduced in quantity, rather than simply interacting with a drug, and the toxicity is reduced. Other proteins that may be labeled with antisense agents for the treatment of lung conditions include, but are not limited to: human adenosine A2a receptor, human adenosine A2b receptor, human IgE b receptor, CD23 antigen of the Fc-epsilon receptor human, human histidine decarboxylase, human beta-tryptase, human tryptase-I, human prostaglandin D synthase, cyclooxygenase-2-human, human eosinophilic cationic protein, human eosinophil-derived neurotoxin, human eosinophil peroxidase, human intercellular adhesion molecule-1 (ICAM -1), human vascular cell adhesion molecule 1 (VCAM-1), human endothelial leukocyte adhesion molecule (ELAM-1), human P-selectin, human endothelial monocyte activation factor, human IL-3, IL-4 human, human IL-5, human IL-6, human IL-8, neutrophil chemotactic factor derived from human monocyte, human neutrophil elastase, human neutrophil factor oxidase, cathepsin G h umano, human defensin 1, human defensin 3, human macrophage inflammatory protein-1-alpha, human muscarinic acetyloline HM1 receptor, human muscarinic actilcholine HM3 receptor, human fibronectin, human GM-CSF, human tumor necrosis factor OI, leukotriene C4 human synthase, human major basic protein, and human endothelin 1. In these latter objects, and in objective genes in general, it is particularly imperative to eliminate or reduce the adenosine content of the corresponding antisense oligonucleotide to avoid its decomposition products of adenosine release. As used herein, the term "treating" or "treatment" of a lung disease refers to a treatment which decreases the likelihood that the subject administered with such treatment will manifest symptoms of lung disease. The term "decreased regulation" refers to inducing a decrease in the production, secretion or availability (and thus a decrease in concentration) of the target intracellular protein.

The present invention relates mainly to the treatment of human subjects but can also be used for the treatment of other mammalian subjects, such as dogs and cats, for veterinary purposes. The target proteins are preferably mammals and more preferably of the same species as the subject being treated. In general, "antisense" refers to the use of small, synthetic oligonucleotides, which are assimilated to DNA of a chain, to inhibit the expression of the gene by inhibiting the function of the target messenger RNA (mRNA). Milligan, J.F. et al., J. Med. Chem. 36 (14), 1923-1937 (1993). In the present invention, the inhibition of the expression gene of the adenosine receptor A- ^ or A ^ is desired. The del-, gene expression is inhibited through hybridization to encode sequences (sense) in a specific messenger RNA (mRNA) by hydrogen binding according to the Watson-Crick base pairing rules. The mechanism of antisense inhibition is that the oligonucleotides applied exogenously decrease the mRNA or protein levels of the target gene or cause changes in the growth characteristics or forms of the cells. Id. See also Helene, C. and Toulme, J., Biochim. Biophys. Acta 1049, 99-125 (1990); Cohen, J.S., Ed., Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression; CRC Press: Boca Ratón, FL (1987). As used herein, "antisense oligonucleotides" is defined as a short sequence of synthetic nucleotides that (1) hybridize to any coding sequence in an mRNA, which encodes the target protein, in accordance with the hybridization conditions described in the following, and (2) after hybridization causes a decrease in the expression of the target protein gene. The adenosine A- or A3 receptor adenosine mRNA sequence is derived from the DNA base sequence of the gene that expresses either the A ^ or A3 adenosine receptor. The sequence of the human genomic adenosine A- ^ receptor is known and is described in U.S. Patent No. 5,320,963 to G. Stiles et al. The adenosine A receptor has been cloned, sequenced and expressed in rats (see F. Zhou et al., Proc. Na t'l Acad. Sci. USA 89: 7432 (1992)) and humans (see MA Jacobson et al. , United Kingdom Patent Application No. 9304582.1 (1993)). In this way, antisense oligonucleotides that regulate the decrease in adenosine A- ^ or A3 receptor production can be produced according to standard techniques. One aspect of this invention is an antisense oligonucleotide having a sequence capable of specifically binding to any sequence of an mRNA molecule which encodes a protein associated with airway disease to prevent translation of the mRNA molecule. Chemical analogs of oligonucleotides (eg, oligonucleotides in which phosphodiester linkages have been modified, for example, for methylphosphonate, phosphotriester, phosphorothioate, phosphorothioate or phosphoroamidate, to carry a more stable oligonucleotide in vivo) are also one aspect of the present invention. Phosphodiester linkages that are naturally found in oligonucleotides are susceptible to degradation by endogenously found cellular nucleases, while many analogue linkages are highly resistant to nuclease degradation. See Milligan et al., And Cohen, J.S., supra. Protection from degradation can be achieved by the use of a "crowned 3-end" strategy, whereby the nuclease-resistant bonds are substituted for phosphodiester bonds at the 3 'end of the oligonucleotide.

See Tidd, D.M. and Warenius, H.M., Br, J. Cancer 60, 343-350. (1989); Sha, J.P. et al., Nucleic Acids Res. 19, 747-750 (1991). All the phosphoroamidate, phosphorothioate and methylphosphonate bonds function properly in this way. The more prolonged modification of the phosphodiester structure has been shown to impart stability and may allow improved affinity and increased cell permeation of oligonucleotides. See Milligan, et al., supra. Many different chemical strategies have been employed to replace the total phosphodiester structure with novel bonds. Id. The structure analogs include phosphorothioate, phosphorodithioate, methylphosphonate, phosphoramidate, borane-phosphate, phosphotriester, acetal, 3'-thiophor acetal, 5'-thioformacetal, 5'-thioether, carbonate, 5'-N bonds. -carbamate, sulfate, sulphonate, sulfamate, sulfonamide, sulfone, sulphite, sulfoxide, sulfur, hydroxylamine, methylene (methylimino) (MMI) or methyleneoxy (ethylimino) (MOMI). Phosphorothioate and methylphosphonate modified oligonucleotides are particularly preferred because of their availability through automated oligonucleotide synthesis. Id. Where appropriate, antisense oligonucleotides can be administered in the form of their pharmaceutically acceptable salts. The antisense oligonucleotides can be of any suitable length (e.g., from about 10 to 60 nucleotides in length), depending on the particular objective required and the mode of delivery thereof. Preferably, the antisense oligonucleotide is directed to a region of mRNA that contains a junction between the intron and exon. Where the antisense oligonucleotide is directed to an intron / exon junction, it can either completely cover the junction or it can be close enough to the junction to inhibit the separation of the exon involved during the processing of the precursor mRNA to the mature mRNA (e.g. , with the 3 'or 5' terminus of the antisense oligonucleotide which is positioned within, for example, 10, 5, 3 or 2 nucleotides of the intron / exon junction). When practicing the present invention, the antisense nucleotides administered may be related to the origin of the species to which it is administered. When treating humans, it can be used if you want human antisense. The pharmaceutical compositions comprise an antisense oligonucleotide as given in the foregoing effective to reduce the expression of an adenosine receptor Aj ^ or A3 by passing through a cell membrane and specifically binding to the mRNA encoding an adenosine receptor A- ^ or A3 in the cells to prevent their translation is another aspect of the present invention Such compositions are provided in a suitable pharmaceutically acceptable carrier (eg, sterile pyrogen-free saline solution) Antisense oligonucleotides can be formulated with a hydrophobic carrier capable of passing through a cell membrane (eg, in a liposome, with the liposomes carried in a pharmaceutically acceptable aqueous carrier.) Oligonucleotides can also be coupled to a substance which inactivates mRNA, such as a ribozyme. can be administered to a subject to inhibit the activation of and adenosine receptors - ^ or A3, in which the subject is in need of such treatment for any of the reasons discussed here. In addition, the pharmaceutical formulation may also contain chimeric molecules comprising antisense oligonucleotides attached to molecules which are known to be internalized by the cells. These oligonucleotide conjugates utilize cell uptake pathways to increase cellular concentrations of oligonucleotides. Examples of macromolecules used in this way include transferrin, asialoglycoprotein (linked to oligonucleotides through polylysine) and streptavidin. In the pharmaceutical formulation the antisense compound can be contained within a lipid particle or vesicle, such as a liposome or microcrystal. The particles can be of any suitable structure, such as unilane or plurilaminar, while the antisense oligonucleotide is contained therein. Positively charged lipids such as N- [1- (2, 3-dioleoxy) propyl] -N, N, N-trimethyl-ammonium methylsulfate, or "DOTAP", are particularly preferred for such particles and vesicles. The preparation of such lipid particles is well known. See, for example, U.S. Patent Nos. 4,880,635 to Janoff et al.; 4,906,477 to Kurono et al .; 4,911,928 from Wallach; 4,917,951 of Wallach; 4,920,016 to Alien et al .; 4,921,757 to Wheatley et al .; etc. The subjects can be administered with the active composition by any means which transports the composition of the antisense nucleotide to the lung. The antisense compounds described herein can be administered to the lungs of a patient by any suitable means, but are preferably administered by generating an aerosol comprised of respirable particles, the respirable particles comprised of the antisense compound, the particles which are inhaled by the subject . The respirable particles can be liquid or solid. The "particles may optionally contain other therapeutic ingredients." The particles comprised of the antisense compound for practicing the present invention should include particles of respirable size: ie, particles of a size small enough to pass through the mouth and larynx after inhalation. and within the bronchus and alveolus of the lungs In general, particles that are in the range of about 0.5 to 10 microns in size are respirable.The non-respirable size particles which are included in the aerosol tend to deposit in the throat and to be swallowed, and the amount of non-respirable particles in the aerosol is preferably decreased.For nasal administration, a particle size in the range of 10-500 μm is preferred to ensure retention in the nasal cavity. liquid of the active compound to produce an aerosol can be prepared combines the antisense compound with a suitable vehicle, such as sterile pyrogen-free water. Other "therapeutic" compounds may optionally be included.Solid particulate compositions containing respirable dry particles of the micronized antisense compound may be prepared by grinding the dry compound with a mortar and pestle, and then passing the chromed composition through a dry sieve. 400 mesh for breaking or separating large agglomerates A solid particulate composition comprised of the antisense compound may optionally contain a dispersant which serves to facilitate the formation of an aerosol A suitable dispersant is lactose, which may be mixed with the antisense compound in any appropriate relationship (for example, a relationship by weight).

Again, other therapeutic compounds may be included as well. The dose of the antisense compound administered will depend on the disease to be treated, the condition of the subject, the particular formulation, the route of administration, the timing of administration to a subject, etc. In general, intracellular concentrations of the oligonucleotide from 0.05 to 50 μM or more particularly from 0.2 to 5 μM are desired. For administration to a subject such as a human, a dose of about 0.01, 0.1 or 1 mg / kg is typically used up to 50; 100 or 150 mg / kg. Depending on the solubility of the particular formulation of the active compound administered, the daily dose may be divided among one or more dose unit administrations. The administration of the antisense compounds can be carried out therapeutically (i.e., as a rescue treatment) or prophylactically. The aerosols of liquid particles comprising the antisense compound can be produced by any suitable means, such as with a nebulizer. See, for example, U.S. Patent No. 4,501,729. Nebulizers are commercially available devices which transform solutions or suspensions of the active ingredient into a therapeutic aerosol vapor either by accelerating a compressed gas, typically air or oxygen, through a narrow venturi orifice or by means of ultrasonic agitation. Formulations suitable for use with nebulizers consist of the active ingredient in a liquid carrier, the active ingredient comprising up to 40% w / w of the formulation, but preferably less than 20% w / w, the carrier is typically water or a solution dilute aqueous alcohol, preferably made isotonic with body fluids by the addition of, for example, sodium chloride. Optional additives include preservatives if the formulation is not prepared sterile, for example, methyl hydroxybenzoate, antioxidants, flavoring agents, volatile oils, buffering agents and surfactants. The aerosols of solid particles comprising the active compound can be produced in a similar manner with any aerosol generator of solid particulate medicament. Aerosol generators for delivering solid particulate medicaments to a subject produce particles which are respirable, as explained above, and generate an aerosol volume containing a predetermined metered dose of a medicament in a ratio suitable for human administration. One illustrative type of the solid particulate aerosol generator is an insufflator. Formulations suitable for administration by insufflation include finely diminished powders which can be delivered by means of an insufflator or taken into the nasal cavity in the form of an inhalation. The insufflator, the powder (for example, a metered dose of the same effective to carry out the treatments described herein) is contained in capsules or cartridges, typically made of gelatin or plastic, which are either cut or opened ip in situ and the powder supplied by extraction of air through the device after inhalation or by means of a manually operated pump. The powder employed in the insufflator consists either simply of the active ingredient or of a powder mixture comprising the active ingredient, a suitable powder diluent, such as lactose, and an optional surfactant. The active ingredient typically comprises from 0.1 to 100 w / w of the formulation. A second type of "illustrative aerosol generator" comprises a metered dose inhaler Metered dose inhalers are pressurized aerosol dispensers, which contain a suspension formulation or solution of the active ingredient in a liquefied vehicle. devices discharge the formulation through a valve adapted to supply a measured volume, typically from 10 to 150 μl, to produce a fine particle mist containing the active ingredient. Suitable carriers include certain chlorofluorocarbon compounds, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane and mixtures thereof. The formulation may additionally contain one or more co-solvents, for example, ethanol, surfactants, such as oleic acid or sorbitan triolate, antioxidants and suitable flavoring agents. The aerosol, whether it is formed of solid or liquid particles, can be produced by the aerosol generator at a rate of about 10 to 150 liters per minute, more preferably about 30 to 150 liters per minute, and more preferably of approximately 60 liters per minute. Aerosols that contain large amounts of medication can be administered more quickly. The following examples are provided to illustrate the present invention, and should not be construed as limiting thereof. In these examples, μM means micromolar, ml means milliliters, μm means micrometers, mm means millimeters, cm means centimeters, ° C means degrees Celsius, μg means micrograms, mg means milligrams, g means grams, kg means kilograms, M means molar and h it means hours.

EXAMPLE 1 Design and synthesis of antisense oligonucleotides The design of antisense oligonucleotides against adenosine receptors A-j, and A may require the solution of the secondary complex structure of the receptor A- ^ target of mRNA and the target A3 receptor of the mRNA. After generating this structure, antisense nucleotides with mRNA target regions are designed, which can be constructed to confer functional activity or stability to the mRNA and which can optimally overlap the initiation codon. Other objective sites are easily usable. As a demonstration of the specificity of the antisense effect, other oligonucleotides not wholly complementary to the target mRNA are included, but which contain identical nucleotide compositions on a w / w basis, as well as controls in the antisense experiments. The A-j ^ adenosine receptor of secondary structure mRNAs is analyzed and used as described above to design an antisense phosphorothioate oligonucleotide. The antisense oligonucleotide which is synthesized was designated HAdAlAS and has the following sequence: 5 '-GAT GGA GGG CGG CAT GGC GGG-3' (SEQ ID NO: 1) As a control, an antisense nucleotide is synthesized. mismatched phosphorothioate designated HAdAlMM with the following sequence: 5 '-GTA GCA GGC GGG GGG GGC-3' (SEQ ID NO: 2) * Each oligonucleotide has identical base content and general sequence structure. Homology investigations in GENBANK (release 85.0) and EMBL (release 40.0) indicate that the antisense oligonucleotide is specific for A- receptor genes, adenosine in rabbits and humans, and that mismatched control is not a candidate for hybridization with any known gene sequence. The A3 adenosine receptor of the mRNA secondary structure is analyzed in a similar manner and used as described above to design two phosphorothioate antisense oligonucleotides. The first synthesized antisense oligonucleotide (HAdA3ASl) has the following sequence: "-GTT GTT GGG CAT. CTT GCC-3" (SEQ ID NO: 3) As a control, an uncoupled phosphorothioate antisense oligonucleotide (HAdA3MMl) is synthesized, having the following sequence: 5 '-GTA CTT GCG GAT CTA GGC-3' (SEQ ID NO: 4) A second phosphorothioate antisense oligonucleotide (HAdA3AS2) is also designated and synthesized, having the following sequence: 5 '-GTG GGC CTA GCT CTC GCC-3' (SEQ ID NO: 5) its control oligonucleotide (HAdA3MM2) has the sequence: 5 '-GTC GGG GTA CCT GTC GGC-3' (SEQ ID NO: 6) The phosphorothioate oligonucleotides are synthesized on an Oligonucleotide Synthesizer from Applied Biosystems Model 396, and purified using NENSORB chromatography (DuPont, MD).

EXAMPLE 2 Test of the Antisense Oligonucleotides in the Adenosine Al Receptor in vitro The antisense oligonucleotide against the human Aβ receptor (SEQ ID NO: 1) described above is tested for efficacy in an in vitro model using HTB-54 lung adenocarcinoma cells. The lung adenocarcinoma cells HTB-54 are shown to express the A- ^ adenosine receptor using standard Northern blotting procedures and receptor probes designed and synthesized in the laboratory. The human lung adenocarcinoma cells HTB-54 (106/100 mm in tissue culture plate) are exposed to HAdAlAS * or 5.0 μM HAdAlMM for 24 hours, with a fresh change of media and oligonucleotides after 12 hours of incubation. After 24 hours of exposure to the oligonucleotides, the cells and their A? ~ It is extracted by standard procedures. A 21-mer probe is synthesized that corresponds to the target mRNA region for the antisense (and therefore has the same sequence as the antisense, but phosphorothioate is not formed) and is used to probe northern blots of prepared RNA of HAdAlAS-treated, HAdAlMM-treated and untreated HTB-54 cells. This analysis of blots clearly shows that HAdAlAS pero. no HAdAlMM effectively reduce the human adenosine receptor mRNA by > fifty%. This result shows that HAdAlAS is a good candidate for an anti-asthma drug since it decreases the intracellular mRNA for the A-j ^ adenosine receptor, which is involved in asthma.

EXAMPLE 3 Efficacy of Anti-Sense Oligonucleotides in the Adenosine Receptor A ^ in vivo A fortuitous homology between rabbit and human DNA sequences within the A- ^ adenosine gene overlapping the start condom allows the use of phosphoroticatc antisense oligonucleotides initially designated for use against the human adenosine A ^ receptor in a model of rabbit. New Zealand white rabbits free of Pasteurella neonates are immunized intraperitoneally within 24 hours of birth with domestic acarid extract (D. farinae) antigen 312 units / ml (Berkeley Biologicals, Berkeley, CA), mixed with 10% chick. Immunizations are repeated weekly for the first month and then every two weeks for the next two months. At 3-4 months of age, eight rabbits sensitized with a mixture of ketamine hydrochloride (44 mg / kg) and acepromazine maieate (0.4 mg / kg) administered intramuscularly are anesthetized and relaxed. The rabbits are rested in a comfortable position on a small, cushioned, animal-molded board and tubed with a 4.0 mm intratracheal tube (Mallinkrodt, Inc., Glens Falls, NY). A 2.4 mm outer diameter polyethylene catheter is passed with a latex balloon attached to the esophagus and the same distance (approximately 16 cm) is maintained from the mouth in all experiments. The intratracheal tube is attached to a Fleisch heat pneumotachograph (size 00, DOM Medical, Richmond, VA), and the flow is measured using a Validyne differential pressure transducer (Model DP-45161927, Validyne Engineering Corp., Northridge, CA) driven by a Gould carrier amplifier (Model 11-4113; Gould Electronic, Cleveland, OH). The esophageal balloon is attached to one side of the differential pressure transducer, and the external flow of the intratracheal tube is connected to the opposite side of the pressure transducer to allow recording of transpulmonary pressure. The flow is integrated to give a continuous tidal volume, and measurements of total lung resistance (LR) and dynamic adaptability (Cdyn) at the isovolumic and zero flow points, respectively, are calculated using an automated respiratory analyzer. (Model 6, Buxco, Sharon, CT). Animals are randomized and obtained in the Day 1 values pretreatment for PC50 by sprayed adenosine. Antisense oligonucleotides (HAdAlAS) or mismatched control (HAdAlMM) are dissolved in sterile physiological saline in a concentration of 5000 ug (5 mg) per 1.0 ml. The animals are subsequently administered with antisense or mismatched antisense oligonucleotides through the intratracheal tube (approximately 5000 μg in a volume of 1.0 ml), twice a day for two days. Are aerosols generated from either saline, adenosine, or oligonucleotides? antisense or decoupled by an ultrasonic nebulizer (DeVilbiliss, Somerset, PA), producing aerosol droplets 80% of which are smaller than 5 μm in diameter. In the first branch of the experiment, four allergic rabbits administered with antisense oligonucleotides and four with mismatched control oligonucleotides are randomly selected. On the morning of the third day, the PC50 values (the concentration of adenosine spray in mg / ml required to reduce the dynamic adaptability of the bronchial airways to 50% of the baseline value) are obtained and are compared with the values PC50 obtained for these animals before exposure to the oligonucleotide. After an interval of 1 week, the animals are crossed, with those previously administered with uncoupled control oligonucleotide now antisense oligonucleotide administered, and those previously treated with the antisense oligonucleotide now administered the uncoupled control oligonucleotides. The methods of treatment and measurements are identical to those used in the first branch of the experiment. It could be indicated that in six of the eight animals treated with the antisense oligonucleotide, adenosine-induced bronchoconstriction can not be obtained up to the solubility limit of adenosine, 20 mg / ml. For the purpose of calculation, PC50 values for these animals are set at 20 mg / ml. The values given therefore represent a minimal figure for antisense efficacy. The real effectiveness is greater. The results of this experiment will be illustrated, in both Figure 1 and Table 1.

TABLE 1, EFFECTS OF ANTI-AGING OLIGONCLEOTIDE IN THE RECEIVER A? OF ADENOSINE AFTER VALUES PC50 IN ASETIC RABBITS.

Control uncoupled Oligonucleotide Antisense in the A- receptor, The results are presented as the mean (N = 8) ± SEM. The significance is determined by analysis of repeated measures of variance (ANOVA), and the Tukey's protected t-test. ** Significantly different from all other groups, P < 0.01.

In both arms of the experiment, the animals receiving the antisense oligonucleotide show an increase in the order of magnitude in the dose of sprayed adenosine required to reduce the dynamic adaptability of the lung by 50%. No effect of the uncoupled control oligonucleotide of PC50 values is observed. No toxicity is observed in any animal that already receives the inhaled antisense oligonucleotide or center! These results clearly show that the lung has exceptional potential as a target for therapeutic intervention based on antisense oligonucleotide in lung disease. They also show, in a model system which closely resembles human asthma, that the diminished regulation of adenosine A- ^ receptor greatly eliminates the adenosine-induced bronchoconstriction between the asthmatic airways. Bronchial hyperresponsiveness in the allergic rabbit model of human asthma is an excellent endpoint for antisense intervention since the tissues involved in this response are closest to the point of contact with the sprayed oligonucleotides, and the model closely simulates a disease important human EXAMPLE 4 Specificity of an antisense oligonucleotide of the adenosine receptor A ^ At the conclusion of the cross experiment of Example 3, the airway muscle of all rabbits is analyzed quantitatively for the number of adenosine A- ^ receptors. As a counterpart for the specificity of the antisense oligonucleotide, A2 adenosine receptors which should not be affected, are also quantified. The uniform muscular tissue of the airways of each rabbit is dissected and a fraction of the membrane prepared according to the methods described (J. Kleinstein and H. Glossmannx Naunyn-Schmiedeberg's Arch. Pharmacol. 305, 191-200 ( 197-8), with slight modifications.The membrane and plasma preparations are stored without purification at -70 ° C until the time of the assay.The protein content is determined by the Bradford method (M. Bradford, Anal. JBiochejn. 72, 240-254 (1976)). Frozen membranes and plasma are thawed at room temperature and incubated with 0. 2 U / ml of adenosine deaminase for 30 minutes at 37 ° C to eliminate endogenous adenosine. The link of [3H] DPCPX (specific to the A ^ or [3H] CGS-21680 receptor (specific to the A2 receptor) as previously described.

Ali et al., J. Pharmacol. Exp. Ther. 268, 1328-1334 (1994); S. Ali et al., Am. J. Physiol 266, L271-277 (1994). As illustrated in both Figure 2 and Table 2, animals treated with the antisense oligonucleotide of adenosine in the crossover experiment have a nearly 75% decrease in the number of A- ^ receptors compared to controls, as assayed by the specific binding of DPCPX specific antagonist A-,. There is no change er. the number A2 adenosine receptors, as tested by the specific binding of 2- [p- (2-carboxyethyl) -phenethylamino] -5 '- (N-ethylcarboxamido) adenosine (CGS-21680) specific agonist to the A2 receptor .

TABLE 2. SPECIFICITY OF THE ACTION OF THE ANTICIPIENT OLIGONUCLEOTIDE OF THE RECEIVER A? OF ADENOSINE Oligonucleotide Antisense control oligonucleotide mismatched for A-, Specific binding to A1 1105 ± 48 * 293 ± 18 Specific binding to A2 302 ± 22 442 ± 171 The results are presented as the mean (N = 8) + SEM. The significance is determined by analysis of repeated measures of variance (ANOVA), and Tukey's protected t-test. ** Significantly different from mismatch control, P < 0.01.

The foregoing demonstrates the efficacy of antisense oligonucleotides in the treatment of diseases of the respiratory tract. Since the anti-sense oligonucleotides described above eliminate the receptor systems responsible for adenosine-mediated bronchoconstriction, it may be less imperative to remove adenosine from them, however, it may be preferable to remove adenosine even from these oligonucleotides. Below are examples of such adenosine-free oligonucleotides in Example 5.

EXAMPLE 5 The method of the present invention is practiced with the following objective antisense oligonucleotides for their corresponding proteins, in essentially the same manner as given above, for the treatment of various conditions in the lungs. A series of target antisense oligonucleotides for the protein mRNA involved in inflammation is described below. Nucleoside adenosine has been eliminated to prevent its release during degradation. In the following, the first sequence provided after the naming of the protein involved in the target inflammation is the antisense sequence that marks the initiation codon, in which the naturally occurring adenosine is replaced by one of the following: (1) a universal base that is not adenosine; (2) an adenosine analog that lacks the ability to bind to Al and / or A3 adenosine receptors; or (3) a "spacer". Any of these three is represented in the sequence as the letter "E", recognized by the IUPAC-IÜE Nomenciacure Commission as "not-A". See Pa tentin User Manual, p.99 (November 1990). Listed below the target antisense sequence against the start codon are additional antisense oligonucleotide sequences directed against other portions of the mRNA of the target protein. These additional sequences are the "de-adenosine antisense sequences", which do not contain adenosines within it. sequence. Fragments of the following sequences that are at least ten, and preferably at least twelve, nucleotides in length are also a feature of the present invention and are useful for carrying out the present invention. The fragments indicated below that extend multiple test lines indicate "5'-" at the beginning of the same, and "-3" at the end thereof.

Human adenosine Al receptor: 5 '-GGC GGC CTG GBB BGC TGB GBT GGB GGG CGG CBT GGC GGG CBC BGG CTG GGC-3 'des-adenosine antisense sequences: TTT TCC TTC CTT TGT CTC TCT TC GCT CCC GGC TGC CTG CTC GGC CGT GCG GCT CTG TCG CTC CCG GT Z- CCG CCC TCC GGG GGG TC TGC TGC CGT TGG CTG CCC CTT CTG CGG GTC GCC GG TGC TGG GCT-TGT GGC "GGC CTC TCT TCT GGG CCT GGT CCC TCC GT GGT GGC TCC TCT GC GCT TGG TCC TGG GGC TGC T G TCT CCT CTC CTT A2a human adenosine receptor: GTBCBCCGBGGBGCCCBTGBTGGGCBTGCCBCBGBCGBCBGGC des-adenosine antisense sequences: HSA2ARECAS1: TGC TTT TCT TTT CTG GGC CTC SEC. FROM IDENT. NO: 7) HSA2ARECAS2: TGT GGT CTG TTT TTT TCT G HSA2ARECAS3: GCC CTG GGG CGC TCT CC HSA2ARECAS4: GCC GCC CGC CTG GCT CCC HSA2ARECAS5: GGB GCC CBT GBT GGG CBT GCC HSA2ARECAS6 r GTG GTT CTT GCC CTC CTT TGG CTG HSA2ARECAS7 : CCG TGC CCG CTC CCC GGC HSA2ARECAS8: CTC CTG GCG GGT GGC CGT TG HSA2ARECAS9: GGC CCG TGT TCC CCT GGG HSA2ARECAS10: GCC TGG GGC TCC CTT CTC TC HSA2ARECAS11: GCC CTT CTT GCT GGG CCT C HSA2ARECAS12: TC-C TGC TGC TGG TGC 7G7 G CCCC Human adenosine A2b receptor: 5 '-BCBGCGCGTCCTGTGTCTCCBGCBGCBTGGCC GGGCCBGCTGGGCCCC-3' des-adenosine antisense sequences: HSA2BRECAS1: 5 '-GGC GCC GTG CCG CGT CTT GGT GGCGGC GG-3' SEC. FROM IDENT. NO: 8) HS-A2BRECAS2: 5 • -GTT CGC GCC CGC GCG GGG CCCCTC CGG TCC-3 'HSA2BRECAS3: 5' -TTG GCC CGC GCG CCC GCC CGTCTC GGG CTG GGC GG-3 'HSA2BRECAS4: CGG GTC GGG GCC CCC CGC GGC C HSA2BRECAS5: 5 '-GCC TCG GGG CTG GGG CGC TGGTGG CCG GG-3' HSA2BRECAS6: CCG CGC CTC CGC CTG CCG CTT CTG HSA2BRECAS7: GCT GGG CCC CGG GCG CCC CCT HSA2BRECAS8: CCC CTC TTG CTC GGG TCC CCG TG A3 Receiver of human adenosine: 5'-BCB GBG CBG TGC TGT TGT TGG GCB TCT TGC CTT CCC BGG G-3 'antisense oligonucleotide sequences: CCC TTT TCT GGT GGG GTG GTG CTG TTG TTG GGC TTT CTT CTG TTC CC ß Receptor of Human IgE: 5 '-BTTTGCTCTCCTBTTBCTTTCTGTGTCCBTTTTTT CBTTBBCCGBGCTGT-3' des-adenosine antisense sequences: HUMIgESrASl: TTT CCC CTG GGT CTT CC SEC. FROM IDENT. NO: 9) HUMIgESrAS2: CTC CTG CTC TTT TTT C CD23 antigen of human Fc-epsilon receptor (Ige receptor) 5 '-TCTCTGBBTBTTGBCCTTCCTCCBTGGCGGTCCTGCTT GGBTTCTCCCGB-3' Des-adenosine antisense sequences: HUMIgErCD23ASl: GCC TGT GTC TGT CCT CCT (SEQ ID NO: 10) HUMIgErCD23AS2: GCT TCG TTC CTC TCG TTC HUMIgErCD23AS3: CTG CTG GGT GCC CTT GCC G HUMIgErCD23AS: GTC CTG CTC CTC CGG GCT GTG G HUMIgErCD23AS5: 5 • -GTC GTG GCC CTG GCT CCG GCTGGT GGG CTC CCC TGG-3 'HUMIgErCD23AS6: CCT TCG CTG GCT GGC GGC GTG C HUMIgErCD23AS7: GGG TCT TGC TCT GGG CCT GGCTGT HUMIgErCD23AS8: GGC CGT GGT TGG GGG TCT TC HUMIgErCD23AS9: GCT GCC TCC GTT TGG GTG GC Human IgE Receptor, subunit a: 5 '-BCBGTBGBGTBGGGGBTTCCBTGGCBGGBGCCBTC TTCTTCBTGGBCTCC-3' and 5 '-TTC BBG GBG BCC TTB GGT TTC TGB GGG BCT GCT BBC BCG CCB TCT GGB GC-3' Des-adenosine antisense sequences: HUMJgErcrASl: GCCTTTCCTGGTTCTCTT ( ID SECTION NO: 11) GTT GTT TTT GGG GTT TGG CTT Human IgE Receiver, Faith epsilon R: 5 '-GBT CTC TGB BTB TTGB CCT TCC BTG GCG GTCCTG CTT GGB-3 'Desen adenosine antisense sequences: HSJGEBFRAS1: GCC TGT GTC TGT CCT CCT. { SEC. FROM IDENT. NO: 12) HSJGEBFRAS2: GCT TCG TTC CTC TCG TTC HSJGEBFRAS3: CTG CTG GGT GCC CTT GCC G HSJGEBFRAS4: GTC CTG CTC CTC CGG GCT GTG G HSJGEBFRAS5: 5 '-GTC CTC GCC CTG GCT CCG GCTGGT GGG CTC CCC TGG-3' HSJGEBFRAS6: CCT TCG CTG GCT GGC GGC GTG C HSJGEBFRAS7: CCC BGB BCG BGB CCC GGB CCG BCB HSJGEBFRAS8: GGC CGT GGT TGG GGG TCT TC KSJG? 3FRAS9: GCT GCC TCC GTT TGG GTG GC Human histidine decarboxylase: 5 '-CTC TGT -CCC TCT CTC TCT GTB CTC CTC BGG CTC CBT CBT CTC CCT TGG GC-3' Des-adenosine antisense sequences: HUMHDCAS1: TCT CCC TTG GGC TCT GGC TCC TCT TC ^ (SEC. DE IDENT NO: 13) HUMMDCAS2: TCT CTC TCC CTC TCT CTC TGT HUMHDCAS3: CGCCTCCGCCCTGGCTGCTGGGGTGGTGGTGC HUMHDCAS4: TTT TGT TCT TCC TTG CTG CC HUMHDCAS5: GCC CCG CTG CTT GTC TTC CTC G Human beta-triptase: 5 '-GGG CCT GGC CTG GGG CBG GGG CCG CGT BGG CGC GGC TCG CCB GGB CGG GCB GCG CCB GCB GCB GCBGBT TCB GCB TCC TGG-3"Antisense sequences of des-adenosine: HUMBTRYPAS1: CTTGCTCCTGGGGGCCTCCTG (SEQ ID NO: 14) HUMBTRYPAS2: GTC CCT CCG GGT GTT CCC GGC Human Triptase-I: 5 '-CCT GGB CTG GGG CBG GGG CCG CGT BGG CGC GGC TCG CCB GGB CGG GCB GCG CCB GCB GCB GCB GGC 7C? GCB TCC TGG CZ? C -3 BBT TCC-3 'Des-adenosine antisense sequences: HUMTRYAS1: CTTGCTCCTGGGGGCCTCCTG (SEQ.ID.NO.:15) HUMTRYAS2: GTC CCT CTG GCT G TT CCC GGC Prostaglandin human D synthase: 5 '-QCC CBG CBG GBC CBG TCC CBT CCB CBG CGT GTG BT * G BGT BGC CBT TCT CCT GCB GCC GBG-3 'Des-adenosine antisense sequences: HUMPROSYNASl: GGTGTGCGGGGCCTGGTGCC (SEQ ID NO: 16) HUMPROSYNAS 2: CCT GGG CCT CGG GTG CTG CCT GT HUMPROSYNAS 3: GCG CTG CCT TCT TCT CCT GG HUMPROSYNAS 4: 5 '-GTC CTC GCC GGG GCC CTT GCT GCC CTG GCT GT -3' HUMPROSYNAS 5: GCC CTG GGG GTC TGG GTT CGGCTGT Human cyclooxygenase-2: 5 '-TGB GCG CCB GGB CCG CGC BCB GCB GCB GGG CGC GGG CGB GCB TCG CBG CGG CGG GCB GGG- 3 'Desen adenosine antisense sequences: HUMCYCL0XAS1: GGGCGCGGGCGBGCBTCGC (SEQ ID NO: 17) HUMCYCLOXAS2: TTT GGG CTT TTC TCC TTT GGT T Human eosinophilic cationic protein: 5 '-CBG BCB BBT TTG GGB BGT GBB CBG TTT TGG BBC CBT GTT TCC CBG TCT CTG BGC TGT GGC-3 'Antisense sequences of des-adenosine: HSECPAS1: CCTCCTTCC TGG TCT GTC TGC (SEQ ID NO: 18) Neurotoxin 'human eosinophilic derivative: 5' -CCC CBB CBG BBG BBG CBG BCB BBT TTG GGB BGT GBB CBG TTT TGG BBC CBT GTT TCC TGT-3 'Des-adenosine antisense sequences: HSEOSDNAS1: GCC CTG CTG CTC TTT CTG CT (SEC DE IDENT NO: 19) HSEOSDNAS 2: TCC CTT GGT GGG TTG GGC C HSEOSDNAS 3: GCT GGT TGT TCT GGG GTT C HSEOSDNAS 4: TTG CTG CCC CTT CTG TCC C HSEOSDNAS 5: TGT TTG CTG GTG TCT GCG C Main basic protein of human eosinophil: GGG GGB GTT TCB TCT TGG CTT T Desen adenosine antisense sequences: TCT CCC CTT GTT CCT CCC C TCT CCT GCT CTG GTG TCT CCT C TTC CCT CCC TCC CCT GCC GTG TTG TCT GTG GGT GTC C GTT TCG CTC TTG TTG CCC TGG GCC CTT CCC TGC TGG Eosinophilic peroxidase "human: 5 '-GCB CCG TCC BGT GBT GGT GCG GTB CTT GTC GCT GCB GCG CTC GGC CTG GTC CCG GBG BGC-3' Antisense sequence of des-adenosine: HSEPAS1: GCGCTCGGCCTGGTCCCGG (SEQ ID NO: 20 ) HSEPAS2: GGG TCT CCT CTT GTT GTT GC HSEPAS3: TTG CGC CTC CTG CTG GGG GT CC HSEPAS4: CTC TGT TCT TGT TTT GGG GGC HSEPAS5: GGG CCC GGC CGT TGT CTT G HSEPAS6: GTT TGG GGG TTT CCG TTG HSEPAS7: GGG TTC TCC TGG CCC GGG CCT TGC CC HSEPAS8: GGC CGT GGT CCC GGC TTC GTT GC HSEPAS9: CCT GTC TCC GTC TCG GCT CTT CTG HSEPAS10: GGG CCT TGC GCT GTC TTT GGT G Molecule-1 of human intercellular adhesion 1 (CAM-1) : 5 '- CGG BGC CTC CCC GGG GCB GGB TGB CTT TTG BGG GGG BCB CBG BTG TCT GGG CBT TGC CBG GTC CTG GGB BCB GBG CCC CGB GCB GGB CCB GGB GTG CGG GCB GCG CGG GCC GGG GGC TGC TGG GBG CCB TBG CGB GGC TGB G-3 'Antisense sequence of des-adenosine: HSICAM1AS1: GCGCGGGCCG3GGGCTGCTGGG (SEQ ID NO: 21) HSICAM1AS2: GGT TGG CCC GGG GTG CCC C HSICAM1AS3: ~ GCC GCT GGG TGC CCT CGT CCTCTGCGGTC HSICAM1AS4: GTG TCT CCT GGC TCT GGT TCC CC HSICAM1AS5: 5 '-GCT GCG CCC GTT GTC CTC TGG GGT _. GGCCTTC-3 'H¿ICAM1AS6: GCT CCC GGG TCT GGT TCT TGT GT HSICAM1AS7: TGG GGG TCC CTT TTT GGG CCT GTT GT HSICAM1AS8: GGC GTG GCT TGT GTG TTC GGT TTC HSICAM1AS9: TGC CCT GTC CTC CGG CGT CCC Human Vascular Cell Adhesion Molecule 1 (VCAM-1) 5 '-CTG BGC BBG BTB TCT BGB TTC TGG GGT GGT CTC GBT TTT BBBB GCT TGB GBB GCT GCB BBC BTT BTC CBB BGT BTB TTT GBG GCT CCB BGG BTC BCG BCC BTC TTC CCB GGC MTT TTB BGT TGC TGT CGT -3 'Desen adenosine antisense sequence: HSVCAM1AS1: CCTCTTTTCTGTTTTTCCC (SEQ ID NO: 22) HSVCAM1AS2: CTC TGC CTT TGT TTG GGT TCG HSVCAM1AS3: CTT CCT TTC TGC TTC TTC C HSVCAM1AS4: CTGTGTCTCCTGTCTCCGCTTTTTTCTTC HSVCAM1AS5: GTC TTT GTT GTT TTC TCT TCC TTG Adhein Molecule of Human Endothelial Leukocyte (ELAM-1): 5 '-BBG TGB GBG CTG BGB GBB BCT GTG BBG CBB TCB TGB CTT CBB GBG TTC TTT TCB CCC -3' Antisense Sequence of des-adenosine: HUMELAM1AAS1: GTTCTTGGCTTCTTCTGTC (SEC DE -, IDENT NO: 23) HUMMEL1AAS2: CGT TGG CTT CTC GTT GTC CC HUMELAM1AAS3: TGT GGG CTT CTC GTT GTC CC HUMELAM1AAS4: CCC TTC GGG GGC TGG TGG HUMELAM1AAS5: GGC CGT CCT TGC CTG CTG G P Human selectin: Desen adenosine antisense sequence: HUMPSELECT1: CTCTGCTGGT TTTCTGCCTT CTGCCC (SEQ ID NO: 24) Human endothelial monocyte activation factor: Des-adenosine antisense sequence: HUMEMAPIIAS1: 5 '-TTT TCT CTT TCG CTT TCT TTT CGTCTCCTGTTCCTCCTTTT- 3 '(SEQ ID NO: 25) HUMEMAPIIAS2: 5' -TTG CTG TTT TTT CTC CTT CTT CTC TCC TTT CTT TTC -3 ' Human IL3: 5'-GGCGGBCCBGGBGTTGGBGCBGGBGCBGGBCGGGCB GGCGGCTCBTGTTTGGBTCGGCBGGBGGCBCTC -3 Desen adenosine antisense sequence: HUMIL3AAS1: 5 '-CTC TGT CTT GTT CTG GTC CTT CGT GGG GCT CTG (SEQ ID NO: 26) -3' HUMIL3AAS2: TGT CGC GTG G GTG CGG CCG TGG CC Human IL3 Receiver: 5 '-GCBGGBGBCBGGGCBGGGCGBTCBGGBGCBGCGT GBGCCBBBGGBGGBCCBTCGGGBBCGCBGCTCCG GBBCGCBGGBCBGBGGTGCC-3 'Antisense sequence of des-adenosine: TCTGGGGTGTCCTG GCCTTCGTGGTTCC TCTTCCTTCGTTTGC CGTCCGCGGGGGCCCCCGGGCCT GGCTGCGCTCCTGCCCCGC CTCTTTCCCGGGCTCTT GCGCTGGGGGGTGCTCC CGTGTGTTTGCGCCCTCCTCCTGGTCGC GCTTGTCGTTTTGG GGCCGGCTTTGCCCGCCTCCC GGCGCCTGGCCCGC-CC TTCCTGGGCTGCGTGCGC GTTCTGTTCTTCTTCCTGGC Human IL4: 5'-GCCGGCBCBTGCTBGCBGGBBGBBCBGBGGGGGB BGCBGTTGGGBGGTGBGBCCCBTTBBTBGGTGTCGB-3 • Des-adenosine antisense fraction: HUMIL4AS1: CTC TGG TTG GCT TCC TTC-3 '(SEQ ID NO: 27) Human IL4 Receiver: 5 • -GTTCCCBGBGCTTGCCBCCTGCBGCBGGBCCBGGCBGCTC BCBGGGBBCBGGBGCCCBGBGCBBBGCCBCCCCBTTGGGBG BTGCCBBGGCBCCBGGCTG-3 • Desen adenosine antisense sequence: TCTGCGCGCCCCTGCTCC CGCCCGGCTTCTCT CGTGTGGGCTTCGG CCCCGCGCCTCCGTTGTTCTC TGCTCGCTGGGCTTG GGTTTCCTGGGGCCCTGGGTTTC TCTGCCGGGTCGTTTTC GGGTGCTGGCTGCG CTTGGTG CT GGGG CT C C GGCGGCTGCGGGCTGGGTTGGG CTTGGCTGGTTCCTGGCCTCGGG CCTCCTCCTCCTCCTC GCTCCCTTTTTCTTCCTCT TCCCTGCTGCTCTC TGCCCTCCCTTCCCTCCTGG GGTGCCTCCTTGGGCCCTGC GGCTGCTCCTTGCCCC CTCTGGGTCGGGCTGGC GGGGCGTCTCTGTGC CTGGCCTGGGTGCC GCCTCTCCTGGGGG GGTGGCTCCCTGTCC CCTTTTCCCCCGGCTCC GTGGGGGCTTTGGC GGGGGTCTGTGGCCTGCTCCTGGGG AGGGGTCTGGGGCCCTC TTTTGGGGGTCTGGCTTG GCCTGGCTGCCTTCC GGGGCCTGCCGTGGGGC TGTCCTCTGTTGCTCCCCTT TGCCTGCTGTCTGG GGTTCCCGCCTTCCCT Human IL5: 5'-GTGGG? 7T7C7GTGGGG? 7GGC? 7? C? CGTBGGC? GCTCCBBGBGCTBGCBBBCTCBBBTGCBGBBGCBTC CTCBTGGCTCTGBBBCG -3 'Des-adenosine antisense sequence: HUMIL5AS1: TCC CTG TTT CCC CCC TTT (SEQ ID NO: 28) HUMIL5AS2: CGT TCT GCG TTT GCC TTT GGC HUMIL5AS3: GTT TTT TGT TTG TTT TCT HÜMIL5AS4: CTC TTC GTC TTT CTT CTC C HUMIL5AS5: CCT CCT GCC TGT GTC CCT GCT CCC C HUMIL5AS6: GAG GGT TTC TGG CTT CCT CTC T HUMIL5AS7: TGT CTC TCT GTC CTT TTG TT HUMIL5AS8: 5 '-TGT TGT GCG GCC TGG TGC TGC CCT GCCCCG GG-3 ' Antisense oligonucleotide of the human IL receptor: 5 '-CTCBGTGGCCCCCBBBBGGBT GBGTBBTBCBTGCGCCBCGBT GBTCBTBTCCTTTTTBCTBTGBGG-3' Antisense sequence of des-adenosine: CCGTGTCTGTCGTGTCT TTCCTTTGCTCTTG GTGTGTCTTTGCTGT GCCCTGCCTCTCTGC Human IL6: 5 '-CTCCTGGGGGTBCTGGGGC? GGG ?? GGCBGCBGGCBBCBCCBGGBGCBGC CCCBGGGBGBBGGCBBCTGGBCCGB BGGCGCTTGTGGBGBBGGBGTTCBT BGCTGGGCTCCTGGBGGGGBGBTBGBGC-3 'des-adenosine antisense sequence: HUMIL6AS1: GCT TCT CTT TCG TTC CCG GTG GGC TCG -. (SEQ ID NO: 29) HUMILACTS2: GTG GCT GTC TGT GTG GGG CGG CT HUMIL6AS3: GTG CCT CTT TGC TGC TTT C HUMIL6AS4: GAT TCT TTG CCT TTT TCT GC Antisense oligonucleotide human IL6 receptor 5 '-GCBCGCCTCTTGCCBCCTCCTGCGCBGGGCB GCGCCTTGGGGCCBGCGCCGCTCCCGGCGCG GCCBGCBGGGCBGCCBGCBGCGCGCBGCCGB CGGCCBGCBTGCTTCCTCCTCGGCTBCCBCT CCBTGGTCCCGCBGBGGCGGBCBGGC-3' antisense sequence des-adenosine: GGGGGTGGCTTCCTGCC GCGTCTCTGGGCCGTCCC GTCCCTCGGCCCCGCGCCGCGCTCGGCTCCTCTCCC TCTGGCCCGGCTC GGGGCGGGGCGGGGCGGTGGGCGGGC GGCGCTGCCCTGCGC GCGGCGCTGGCCCC TGCTGGCCGTCGGCTGCGCGCTGCTG GCTGCCCT GCTGGCCGCGCCGGG GCCTGTCCGCCTCTGCGGG CGCTGTCTCCTGGC TTGTCTTCCGGCTCT TCTGCTGGGGTGGG GCTGGGCGGCCGGCCCGGT G CTGGGG CT C CT CGGGGGG GGGGGCTCTTCCGG GCTGTCTCCCTCCGGG GCGGGGGTTTCTGGCC GTGGGGGTCTTGCC TGGCCTCCGGGCTCC TGCTTGTCTTGCCTTCCTTC TCTGGTCGGTTGTGGCTCG GGGCTCCGTGGGTCCCTGGC GCCCGTTTGTGTTTTGTC TTTTCCCCTGGCGT CCCTGTGCCCCTCTCCTCTCCTTCCT CTGCTTCTC GCTCTCCTTTGTGGG GCCCTCCCTGCTGCT CTTGGTTTTGGGCT GTGCGTGGGCCTCC Neutrophil chemotactic factor derived from human monocyte: 5 '-GGGGTGGBBBGGTTTGGBGTBTGTCTTTBTGCBCTGB CBTCTBBGTTCTTTBGCBCTCCTTGGCBBBBCTGCBC CTTCBCBCBGBGCTGCBGBBBTCBGGBBGGCTGCCBB GBGBGCCBCGGCCBGCTTGGBBGTCBTGTTTBCBCBC BGTGBGBTGGTTCCTTCCGG-3 'Des-adenosine antisense sequence: HSMDNCFAS1: GCT TGT GTG CTC TGC TGT CTC T (SEQ ID NO: 30) HSMDNCFAS2: 5' -TGG TTC CTT CCG GTG GTT TCT TCC TGG CTC TTG TCC T -3 • HSMDNCFAS3: TTC TCT CCC TGG TTG GC of human neutrophil elastase (medullasin): 5 • -GGGCTCCCGCCGCGBGBGGTTBTGGGCTCCCBGGBCCBC CCGCBCCGCGCGGBCGTTTBCBTTCGCCBCGCBGTGCGC GGCCGBCBTGBCGBBGTTGGGCGCBBTCBGGGTGGCGCC GCBGBBGTGGCCTCCGCGCBGCTGCBGGGBCBCCBTGBB GGGCCBCGCGTGGGGCCGCGCTCGCCGGCCCCCCBCBBT CTCCGBGGCCBGCGCGGTGCCCCCCBGCBGCBBGGCCGG GGTCBTGGTGGGGCTGGGGCTCCGGGGTCTCTGCCCCTC CGTGC-3 'oligonucleotide antisense des-adenosine: HSMEDURAS1: 5 • -TGG TGG GGC TGG GGC TCC GGG GTC CCT GCC CCG TCT TGC-3' (SEQ ID NO:.. 31) HSMEDURAS2: CGC GGG GTG CCG CGC TCG CCG - GCCCCCC HSMEDURAS3: CCT GCC GGG TGG GCT CCC GCC GCG HSMEDURAS4: CGC CGG CCT GCC GGC CCC TC HSMEDURAS5: 5 '-GTG GGT CCT GCT GGC CGG GTC CGG GTC CCG GGG GTG GGG-3' HSMEDURAS6: CGC GBG TCG GCG GCC GBG GGT C Human neutrophil oxidase Factor: 5 • -CGGGBGTGGGGGTCCTGGBCGGCBCTGBBGGCBTCCBGGG CTCCCTTCCBGTCCTTCTTGTCCGCTGCCBGCBCCCCTTC BTTCCBGBGGCTGBTGGCCTCCBCCBGGGBCBTGBTTBGG TBGBBBCTBGGBGGCC-3 • antisense sequence of des-adenosine: HUMN0XFAS1: GGC CTC CBC CBG GGB CBT G (SEQ ID NO:.. 32) HUMN0XFAS2: CTT GTC TGC CTT GTC CGC C HUMNOXFAS3: TCT CTG GGG TTT TCG GTC TGG GTG GHUMNOXFAS4: GCT TTC CTC CTG GGG CTG CTG CTG GGTG-3 'HUMNOXFAS6: CTC CTT CTG CTG CTT CTCC HUMNOXFAS7:' CTT TGG TGT CTT TGT TTT GT HUMNOXFAS8: 5 '-GGCCTCCBCCBGGGBCBTGGTCCTTCTT GTCCGCTGCC -3' Human cathepsin G: 5V '-CCCTCCBCBTCTGCTCTGBCCTGCTGGBCTCTG GBTCTGBBGBTBCGCCBTGTBGGGGCGGGBGTG GGGCCTGCTCTCCCGGCCTCCGBTGBTCTCCCCT GCCTCBGCCCCBGTGGGTBGGBGBBBGGCCBGCB GBBGCBGGBGTGGCTGCBTCTTTCCTG -3' Antisense sequence of des-adenosine: HUMCTHGAS1: GTG GCT CCT GGG GCC TCC CTC GCC G (SEQ ID NO:.. 33) HUMCTHGAS2: TGTGTTGCTGGGTGTTTTCCCGTCTCTGG HUMCTHGAS3: TCT GCC TTC GGG GGT CGT Human Defensin 1: 5 '-CCGGGGCTGCBGCBBCCTCBTCBGCTCTTGCCT GGBGTGGCTCBGCCTGGGCCTGCBGGGCCBCCB GGBGBBTGGCBGCBBGGBTGGCGBGGGTCCTCB TGGCTGGGGTCBCBGBTCCTCTBGCTBGGCBGG GTGBCCBGBGBGGGC-3 • Antiser.ti? C sequence of des-ad rC? Ir.a: HUMDEF1AAAS1: GGG TCC TCG TGG CTG GGG (SEQ ID NO: 34) HUMDEF1AAAS2: GCC TGG GCC TGC BGG GCC HUMDEF1AAAS3: GCT CTT GCC TGG BGT GGC TC HUMDEF1AAAS4: GCC CBG BGT CTT CCC TGG T Human defensin 3: 5 '* - CGCTGCBBTCTGCTCCGGGGCTGCBGCBBCCTCBTC BGCTCTTGCCTGGBGTGGCTCBGCCTGGGCCTGCBG GGCCBCCBGGBGBBTGGCBGCBBGGBTGGCGBGGGT CCTCBTGGCTGGGGTCBCCTGGBGGBGGGBGBGCBGG-3' Antisense sequence of des-adenosine: HUMNTRIIIAS1: GGG GGG TCC CTG TGG TC TCB: HUMNTRIIIAS2: CCT CTC TCC CCT CGT Protein- (SEQ ID NO 35..) Inflammatory 1-alpha human macrophage: RANTES RECEPTOR 5 '-GBGGGGGCBGCBGTTGGGCCCCBBBGGCCCTCTCGT TCBCCTTCTGGCBCGGBGTTGCBTCCCCBTBGTCBB BCTCTGTGGTCGTGTCBTBGTCCTCTGTGGTGTTTG GBGTTTCCBTCCCGGCTTCTCTCTGGTTCCBBGGGB-3 'Desen adenosine Antisense Sequence: K "MRANTAS1: Z-ZZ 777 GTT 7C7 C-GG C7C GTG CC (SEQ ID NO: 36) HUMRANTS2: CCB TCC CGG CTT CTC TCT GGT TCC HUMRANOS'3 : GTC CTCTGT GGT GTT TGG HUMRANTS4: 5 '-CCC TGC TTC CTT TTG CCT GTT TCTTTGTTT CTGGGCTCGT GCC -3' RANTES: 5 -GGGCBCGGGGCBGTGGGCGGGCBBTGTBGGC BBBGCBGCBGGGTGTGGTGTCCGBGGBBTBTGGG GBGGCBGBTGCBGGBGCGCBGBGGGCBGTBGCBB TGBGGBTGBCBGCGBGGCGTGCCGCGGBGBCCTTC BTGGTBCCTGTGGBGBGGCTGTCGGBGG-3 'Des-adenosine antisense sequence: GGGTGTGGTGTCCG CTTGGCGGTTCTTTCGGGTG TTTCTTCTCTGGGTTGGC CTGCTGCTCGTCGTGGTC GTCCGCTCCCGGGTTC GTCTCGCTCTGTCGCCC CTTCCTTCCTTGTC GTGTTCCTCCCTTCCTTGCCTCT Human muscarinic acetylcholine HM1 receptor: Des-adenosine antisense sequence: HSHM1AS1: GTT C? 7 GGT GGC 7? G G7G GGG C (SEQ ID NO: 37) HSHM1AS2: GCT GCC CGG GGT GTG CGC TTG GC HSHM1AS3: GCTCCCGTG CTC GGT TCT CTG TCTCCCGGT HSHM1AS4: CCC CCT TTG CCT GGC GTC TCG G HSHM1AS5: GCC TTC GTC CTC TTC CTC TTC TTC CTTCC HSHM1AS6: 5 * -GCT CCG TGG GGG CTG CTTGGTGGG GGCCTG TGC CTC GGG GTC C-3 'HSHM1AS7: CGG GGC TTC TGG CCC TTG CC Human Muscarinic Acetylcholine HM3 Receptor: Des-adenosine antisense sequence: HSHM3AS1: GGG GTG GGT BGG CCG TGT CTG GGG (SEQ ID NO: 38) HSHM3AS2: GTT GGC CBT GTT GGT TGC C HSHM3AS3: TCT TGG TGG TGC GCC GGG C HSHM3AS4: 5 '-GCG TCT TGG CTT TCT TCT CCT TCG GGC CCT CGG GCC GGT GCT TGT GG-3' HSHM3AS5: 5 '-GCT CCT CCC GGG CGG CCT CCC ~ CGG GCG GGG GCT TCT TG-3' HSHM3AS6: GCG CTG GCG GGG GGG CCT CCT CC HSHM3AS7: 5 '-GCT CTG TGG CTG GGC GTT CCT TGG TGT TCT GGG TGG C-31 HSHM3AS8: TGG CGG GCG TGG TGG CCT CTG TGG TGG HSHM3AS9: GGG CCC GCG GCT GCE GGG G HSHM3AS10: TTG CCT GTC TGC TTC GTC HSHM3AS11: CTT TGC GCT CCC GGG CCG CC Human fibronectin: Desen adenosine antisense sequence: HUMFNA / HSFIB1AS1: CGG TTT CCT TTG CGG TC (SEQ ID NO: 39) HUMFNA / HSFIB1AS2: TTG GCC CGG GCT CCG GGT G HUMFNA / HSFIB1AS3: CCC GCC CGC CCG CCG GCC GCCGC HUMFNA / HSFIB1AS4: 5 '-CCC GCC GGG CTG TCC CCG CCC CGC CCC-31 HUMFNA / HSFIB1AS5: GGC CCG GGG CGC GGG GG HUMFNA / HSFIB1AS6: CGG CCC TCC CGC CCC TCT GG HUMFNA / HSFIB1AS7: GCC GGC GCG GGC GTC GG HUMFNA / HSFIB1AS9: 5 '-CCG CTC GCG CCT GGG GTT CCC TCT CCT CCCCCTGTGC-3' HUMFNA / HSFIB1AS10: GCC TGC CTC TTG CTC TTC HUMFNA / HSFIB1AS11: TGC GTC CGC TGC CTT CTC CC HUMFNA / HSFIB1AS12: CTC TCC TCG GCC GTT GCCTGTGC HUMFNA / HSFIB1AS13: 5 '-TGT CCG TCC TGT CGC CCT TCC GTG GTG C-3' HUMFNA / HSFIB1AS14: TGT TGT CTC TTC TGC CCT C KUMFKA / HSFIB1AS15: GGT G7G C7G GTG CTGGTGGTGGTG HUMFNA / HSFIB1AS16: CCT CTG CCC GTG CTC GCC HUMFNA / HSFÍB1AS17: CTG CCT GGG CTG GCCTCTTCGGGT HUMFNA / HSFIB1AS18: 5 '-GTG GCT TTG GGG CTC TCT TGG TTG CCC TTT- 3' HUMFNA / HSFIB1AS19: 5 '-CTT CTC GTG GTG CCT CTC CTC CCT GGC TTG GTC GT-3 'HUMFNA / HSFIB1AS20: TGT CTG GGG TGG TGCTCCTCTCCC HUMFNA / HSFIB1AS21: TTT CCC TGC TGG CCG TTT GT HUMFNA / HSFIB1AS22: CCT GTT TTC TGT CTT CCT CT HUMFNA / HSFIB1AS23: TTC CTC CTG TTT CTC CGT HUMFNA / HSFIB1AS24: 5 »-TTG GCT TGC TGC TTG CGG GGC TGT CTC C-3 'HUMFNA / HSFIB1AS25: CTT GCC CCT GTG GGC TTT CCC HUMFNA / HSFIB1AS26: TGG TCC GGT CTTCTCCTTGGGGGTC HUMFNA / HSFIB1AS27: GCC CTT CTT GGT GGG CTG HUMFNA / HSFIB1AS28: GCT CGT CTG TCT TTT TCC TTCC HUMFNA / HSFIB1AS29: 5 »-TGG GGG TGG CCG TTG TGG GCG GTG TGG TCC GCC T-3 'HUMF-C .-- /: -: S ?: E1.-_? 30: TGC CZZ TGC 7GG 7C7 77C Human interleukin-8: 5 '-GBTGTTTGTTBCCBBBGCBTCBBGBBTBGCTTTGC TBTCTBBGGBTCBCBTTTBGBCBTBGGBBBBCGC TGTBGGTCBGBBBGBTGTGCTTBCCTTCBCBCBG BGCTGCBGBBBTCBGGBBGGCTGCCBBGBGBGCC BCGGCCBGCTTGGBGTCBTGTTTBCBCBCBGTGBG-3' antisense Se'cuencia des-adenosine: HUMIL8AAS1: GTG TGG CTT CTC CGG TTT (SEQ ID NO:.. 40) HUMIL8AAS2: GCT TGT GTG CTC TGC TGT CTC TG HUMIL8AAS3: 5 '-TTC CTT CCG GTG GTT TCT TCC TGG CTC TTG TCC T-3 * HUMIL8AAS4: TTC TCT TGG CCC TTG GCC C Human IL-8 alpha receptor 5 '-BCBGGGGCTGTBBTCTTCBTCTGCBGGTGGCB TGCCBGTGBBBTTTBGBTCBTCBBBBCCCTCBT CTGTGGBTCTGTBBTBTTTGBCBTGTCCTCTTC; BGTTTCBGCBBTGGTTTGBTCTBBCTGBBGCBCCG GCCBGG-3 'Des-adenosine antisense sequence: TGGCTCGGTGCTTCTGCCCC TGTTGTTGCGGCGCTC GG77GG7G7GGCCCCTG TGGTGCTTCGTTTCC CCCTCTTTCTCTTTGTTC GGGGGTTCTTGTGGC GGGCTGCTTGTCTCGTTCC Human GM-CSF: 5'-CTTGBGCBGGBBGCTCTGGGGCBGGGBGCTGGCBG GGCCCBGGGGGGTGGCTTCCTGCBCTGTCCBGBGT GCBCTGTGCCBCBGCBGCBGCTGCBGGGCCBTCBG CTTCBTGGGGCTCTGGGTGGCBGGTCCBGCCBTGG GTCTGGGTGGGGCTGGGCTGCBGGCTCCGGGC-3 'Antisense sequence of des-adenosine: HUMGCSFAS1: GGT GGT CTG CCB BTG GCC GG (SEQ ID NO:.. 41) HUMGCSFAS2: GGC TGG GGC GCT TCC GCB GG HUMGCSFAS3: GCG GGC GGG TGC GGG CTG CGT GCT GGG HUMGCSFAS4: GGC TGC CCC GCA GGC CCT GC Human tumor necrosis factor a: 5 '-CBCCGCCTGGBGCCCTGGGGCCCCCCTGTCTTCTTGGG GBGCGCCTCCTCGGCCBGCTCCBCGTCCCGGBTCBTGCTTT CBGTGCTCBTGGTGTCCTTTCCBGGGGBGBGBGGG-3 'Antisense sequence of des-adenosine:.-._ .. * :.' • _-_: _: _r'_ I -: - '-r > J ILU .r _ - -T (SEQ ID NO: 42) HSTNFAAS2: GTG CTC BTG GTG TCC TTT CC HSTNFAAS3: GCC CTG GGG CCC CCC TGT CTT CTT GGGG HSTNFAAS4: CCT CTT CCC TCT GGG GGC CG HSTNFAAS5: TCT CTC TCC CTC TCT TGC GTC TCT C HSTIÍFAAS6: TCT TTC TCT CTC TCT CTT CCC C HSTNFAAS7: TTT CCC GCT CTT TCT TCT TC HSTNFAAS8: GGT TGC TGT TTT TCT TCC TCC HSTNFAAS9: T GCT GGC CTG CTT CTG CGT TCTT HSTNFAAS10: GGC CTG TGC TGT TCC TCC HSTNFAASll: TCC GGT TCC TGT CCT CTC TGT CTG TC HSTNFAAS12: GCC CCC TCT GGG GTC TCC CTC TGG C HSTNFAAS13: GTG GTG GTC TTG TTG CTT HSTNFAAS14: GGG CTG GGC TCC GTG TCT C HSTNFAAS15: CBG TGC TCB TGG TGT CC HSTNFAAS16: GCT GBG GGB GCG TCT GCT GGC Human leukotriene C4 synthase: 5 '-CTCGGTBGBCGCGCTCGBBCTCGGGTGGGCCGGTGGTG BGCGGCGGCGBCBCGCGGBBGGCCCTGCGCGCCGBGBTCBC CTGCBGGGBGBBGTBGGCTTGCBGCBGGBCTCCCBGGBGGG TGBCBGCBGCCBGTBGBGCTBCCTCGTCCTTCBTGGTBCCG TCGG7G7GGTGGCBCGGGC7G7G7G7G BGGCG GC7GG-2 'Antisense sequence of des-adenosine: TCT CCG GCT GCC HSU11552AS1 CCT GCT CGT GCC G (SEQ ID NO:.. 43) HSU11552AS2: 5'? -CCT CCT CGT CGT TCA TGG TAC CGGTGT GGT GGC-3 'HSU11552AS3: CTC GGG TGG GCC GGT GGT G HSU11552AS4: GGG CGC GCG CGC TCG CGT HSU11552AS5: 5 • -GGC TCC GGC TCT TCT TTC CCG GCTCCG TCG GCC CGG GGG CCTTGGTCT C-3' HSU11551AS6: CCT CGT CCT TCB TGG TBC CG Human endothelin-1: 5 '-BCCGGCGGBGCCGCCBGGGTGGBCTGGGBGTGGGTT TCTCCCCGCCGTTCTCBCCCBCCGCGCTGBGCTCBGCGC CTBBGBCTGCTGTTTCTGGBGCTCCTTGGCBBGCCBCBBBCBGCBGBGBGBGBBBBTCBTGBGCBBBTBBTCCBTTCTGB BBBBBBGGGBTCBBBBBCCTCCCGT-3 'Des-adenosine antisense sequence: CCCGTTCGCCTGGCGC GCGCTGCGGGTTCCTC GTGGGTTTCTCCCCGCCGTTCTC CGGTCTGTTGCCTTTGTGGG CTTCTTGTCTTTTTGGCT G7TC7TTTCCTGCTTGGC GTCTTTTCCTTTCTT TGTGCTCGGTTGTGGGTC CGCTGGTCCTTTGCC CTGTGTGTTTCTGCTG Antisense oligonucleotides of ET-P endothelin receptor 5 '-GCCCTGTCGGGCGGGBBGCCTCTCTCCTCTCCCCBG BÍCCGCGBCBGGCCGCBGGCBBGBBCCBGCGCBBCCBGG GCGCGTCCGCBCBGBCTTGGBGGCGGCTGCBTGCTGCTB CCTGCTCCBGBBGCGTCCGGTGGCCGCCGC-3' Antisense sequence of des-adenosine: GCGTCCGGTGGCCGCCGC GCCTCTCTCCTCTCCCC GTGGCCCTGTCGGGCGGG TCCTGCCGTCCTGTCTCCTTT TCTTTTGCTGTCTTGT CTTCCCGTCTCTGCTTT Antisense Oligonucleotides endothelin ETA receptor 5 '-CBTCCBCBTGBTTGCTTBGBTTTGTGCTGTBTCTCTCB GGBTTBTCBCTGBTTBCBCBTCCBBCCBGTGCCBGCCBBBB GGBTGCCCTGBGGCBBBGGGTTTCCBTCTTGBGGCBBBTTT GBGGB-3' Antisense sequence of des-adenosine: GTCTGTCCTCCCCGTCTCCTCCC ACTGCTTCTCCCGGGG n * - ** "* -" * t * r * r * ( ~ * * d **! * T f "* GGGTGGCCGGTGTCCCGGGCTCCGGCGCGGCGGC GGCTTCGGCTGC GGGTGGGTGGCGCGG GCTGCCGGGTCCGCGCGGCGCCTGGGCC CTTGTGCTGCTTTT TGCTTGTTCCGTTC TGG? TGCTCCGGTCTGTGTTGTGGTTGTTTTG TTTCTTCTTGGGTGTGGG CCTTGCGGTTTTGG CTGTGGGCCCTTTG GGGCCTTGGCTTCTGGCTC Antisense oligonucleotide of substance P 5 '-CTGCTGBGGCTTGGGTCTCCGGGCGBTTCTCTGCBGBBGBT GCTCBBBGGGCTCCGGCBGTTCCTCCTTGBTCTGGTCGCTGTCG TBCCBGTCGGBCCBGTBBTTCBGBTCBTCBTTGGCTCCTBTTTC TTCTGCBBBCBGCTGBGTGGBGBCBBGBBBBBBGBCTGCCBBGG CCBCGBGGBTTTTCBTGTTGGBTTTTGCGBCGGBCBGTCCCGCG GGGTGCTGAGTTTCTCTGGTTCCTCCGBGCGCB- 3 'Antisense sequence of des-adenosine: CGTGGTCGCTCCGC TTT CT CTGGTTC CT CCG GTCCCGCGGGGTGCTG TCTGGTCGCTGTCGT GTTTCCTTCCTTTTCCGC Antisense oligonucleotide of substance P receptor 5 '-GGCTBBGBTGBTCCBCBTCBCTBCCBCGTTGCCCBCCBCB GBGGTCBCCBCBBTGBCCGTGTBGGCBGCTGCCCBBBGGBCBB TTTGCCBGGCTGGTTGCBCGBBCTGBTTGGGTTCCGBGGTGTT BGTGGBGBTGTTTGGGGBGBGGTCTGBGTCCBCCGGGBGGBCG TTBTCCBTTTCGBBGCTBGGCGGTBBBGCCCTBCTBTCTGTBC BCBBCCCCCCTCTGCBGCBGBGTCCTGTCGTGGCGCCTGGGGC TCBGGGTCC-3 'Antisense sequence of des-adenosine: GTCCTGTCGTGGCGCCTGGGGCTC TTCTTTTGTGGGCT CTTTGGTGGCTGTGGCTG TGGTCTCTGTGGTTG CTGCCCTGGGTCTGG GGGTGTGGCCTTGGGGCCGTCCTCTGGCTCCTCCTCGTGGGCCCCC Cimasa 5 '-GGBGCTGBTBCTGCBGATTTCBGBGGGBBGBBCCCT GBTBCTCBCCBGCTTCBGCTCTGGBGCBCBBGBGBBBGB GCBGCBGGGGGBGBGGBBGBBGCBGCBTCTTCCCBGBGB GGCTGCCTGBGCBBBTGCTGGTTTTCCTTTCCBGTCTTG GGTTTTBTBBCTCCCBGBBGGCBBGBGBGGGGCBBGG-3 'Antisense sequence of des-adenosine: - A i i ILi i ict. TGCTGGTTTTCCTTTCC TGGCAGTGGGTGGGGGTGGGGGTGGGGTGGC TTCCTTGTTCCTGGGGGTGTCCT CTTGCTCTGGGCTTTTCT CCCCTTTTCCTTCC TGTCTGTTTTCCTGGGG CTCT.CCTCTGTCTCTGTGT CCÍTGCCCTGGCCC TCTTCCCTCTCCTGTCTCCTGT CCCTGTGTTCCGCCC GTCTTCCCTCTCCTG ACCTCCTTTTCCTCCG CTGGGTGGGGCCCTG CCTGTTCTCTGCTCCC TGGCTTGGGGTTTCTTCTG TGTGTCTTCTTCCTCTGTT GGCTGGCTTTCTCCTTC TTTTGTCTTCCTGGG TGCCCCTTCTTCCTTTCTTGGG TCCTTGGTGCTTGGGCTGGG endothelial nitric oxide synthase of 5 '-GCGTCTTGGGGTGCBGGGCCCBTCCTGCTGCGCCTGGGCG CTGBGGGTGTCBTBGGTGBTGCTCCCCBCCTCCCBGTTCTTCB CBCG3GGGE3CT7GGGCCCCTCTGGGGGC GGG773GCGGGGE GCTCGGGGGGCTGTGTTCTGGCGC7GG7GGGBGT3GGGBTGCT GGGGCCCGGCTGGGCTCBGGGGCCGGGGTGGCTGGGCCCTGCT TGCCGCBCBGCCCBBGGCCCBGCCCCBGCCCCBGGCCGCBGGG TGGCCCBGGCTCCTGGGCCBCGCTCTTCBBGTTGCCCBTGTTB CTGTGCGTCCGTCTGCTGGBGCBGGC3GCBGBGTGGGBBTTC-3 'Desen adenosine antisense sequence: CTGTGCGTCCGTCTGCTGG GGGGCCGGGGTGGCTGGGCCCTGCTTGCCGC ACGACCCCGGGCCGACCCGAG GCTCGGGGGGCTGTGTTCTGGCGCTGGTGGG CTTGGGCCCCTCTGGGGGCTGGGTT TCCTGCTGCGCCTGGGCGCTG GCGTCTTGGGGTGC GGGGCCGGGGGGCCGGGGG GCCGCTGTTCGTGGGCCTGGG GGTGCCTGTGGCTGCC GGTTGCCCCGGTTGGTGGC GCCGTCCTGCTGCCGGT CGTTGGCTGGGTCCCCCCGC CCGTTTCCTGGGGTCC GCGTGGGGTGCTCC GGTTCCTCGTGCCG CTGCTGCCTTGTCTTTCC GGCCGTGGCGGCGTGGTGGTCC GCCCCCCC7GGCC7TCTGC7C GGGGTCTGGCTGGT TGCCGGTGCCCTTGGCGGC GGTCTTCTTCCTGGTG GCTCTGGGCCCGGCCGGTCTCGG GCGTCTCGTGTTCG CTCTTGTGCTGTTCCGGCCG CTCCTTCCTCTTCCGCCGCC GCCGCTCCCCGCCC GCTCGTCGCCCTGGCCC GGCCTCCTCCTGGCCGC TGTCTCGGGCGGCGGCCTTGGC GCTCCGTTTGGGGCTG CCTCTGGCGCTTCC GGCCCTCGGCCTGGGCGCTC TCTTCCGCCTGTGC TGGTGGCCCTCGTG GCCCCTCCTGGCCTCCGGTGTCC TGTGGTCCCCCGGCTGGT GGCCGGGCCGGTTGGGCGGGC GTGGGCGCCGGCGGGTCCTCC GGGCTGCC CTT CT CC GCCGGGGGTCCCGC GCTCCTGCTGTTCCCTGGGCTCTTCTGCC TCTCTCCTGGGTGGGTGCTGGGTGCCG GGGTCTCCGGGCTTG CCCCGCGCTGCTGGGCGTTCTGC GGTCTTGGGGTTGTC TGTGGCCCCGCTCG TGTCGCCCTCCGTCGCC CGTCGCCGGCCTCGTCC CCTCCTGGGTGCGC GGCGGGCTGGTCCT GGCGTTTTGCTCCTTCCTGG 'Inducible Synthase Nitric Oxide -CTGCCCCBGTTTTTGBTCCTCBCBTGCCGTGGGGBGGB CBBTGGGGTTGCBTCCBGCTTGBCCBGBGBTTCTGGBG BCTTCTTTCCCGTCTCCBCGBGGGGCTGCGGGGBCTCB TTCTGCTGCTTGCTGBGGTTGTGB TB CTGB GGTCB T CC TGTGTCBCTGGBCTGGBGGTGGCBCBGGGGGCTTTCTC CBCBTTGTTGTTGBTGTCTTTTTCCCCBTTCBTTGCBT BCTGGTGGBBTTTGGTCTTGBBCBGBBBTTTCCBBGGB CBGGCCBTCTCTBTGGCTTTBCBBBGCBGGTCBCTTBT GTCBCTTBTCTGGBTTTGBGCTCBGBTGTTCTTCBCTG TGGGGCTTGCBGCTGGCTGCBCTGCCTCCCCGGGGTB-3 ' Human major basic protein: GTTTCATCTT GGCTTTATCC (SEQ ID NO: 44) EXAMPLE 6 A part returning to Figure 3, two asthmatic rabbits are administered with adenosine, and two rabbits are administered with AMpD, at the indicated concentrations, by inhalation as described above in Example 3 », The results (shown in Figure 3 as change of dapatability) indicates that AMpD, a breakdown product of antisense oligodeoxynucleotides containing adenosine, is potent induction of bronchoconstriction, such as adenosine in the hyper-responsive airways of asthmatic rabbits.

EXAMPLE 7 It is found that 21-mer antisense, aerolized phosphorothioate 50% adenosine and 50% guanine plus cytosine in a randomized configuration produces potent bronchoconstrictor effects in hyperreactive airways of asthmatic rabbits as illustrated in Figure 4. The control molecule used in this study, ODN antisense 21-mer phosphorothioate consisting of 50% guanine and 50% thymidine plus cytokine (ODN des-adenosine) does not produce any bronchoconstrictor or any other effect in these rr.isrr.esrr.es ~ .ales. In this study, the bronchoconstrictor effects are measured as a percentage change in bronchial adaptability. Each group consists of two allergic rabbits, and the data shown are for the period following the second of two daily administrations of 5 mg ODN aerolyzed by the nebulizer. These results indicate that antisense oligonucleotides, even when modified to achieve slow degradation, produce adenosine metabolites capable of potentiating bronchoconstriction when administered in asthmatic airways. The examples mentioned above are illustrative of the present invention, and are not constructed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.

LIST OF SEQUENCES (1. GENERAL INFORMATION: (i) APPLICANT: Nyce, Johnathan W. (ii) TITLE OF THE INVENTION: Method for the Treatment of Lung Diseases Using Antisense Oligonucleotides (iii) SEQUENCE NUMBER: 44 (iv) ADDRESS FOR CORRESPONDENCE: (A) - ADDRESSEE: Kenneth D. Sibley (B) STREET: Post Office Drawer 34009 (C) CITY: Charlotte (D) STATUS: NC (E) COUNTRY: USA (F) POSTAL CODE (ZIP): 28234 (v) READING FORM ON THE COMPUTER: (A) TYPE OF MEDIUM: Soft disk (B) COMPUTER: compatible with an IBM PC (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: PatentIn Relay # 1.0, Version # 1.30 (vi) DATA OF THE CURRENT APPLICATION: (A) APPLICATION NUMBER: (B) DATE OF SUBMISSION: (C) CLASSIFICATION: ; -.- üij: N? ORKACID: C OF THE APODEEAEC / AGENT (A) NAME: Sibley, Kenneth D. (B) REGISTRATION NUMBER: 31,665 (C) REFERENCE / NUMBER OF FILE: 5218-32 (ix) TELECOMMUNICATION INFORMATION (A) TELEPHONE: (919) 881-3140 (B) -. TELEFAX: (919) 881-3175 (C) TELEX: 575102 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 1: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 1: GATGGAGGGC GGCATGGCGG G 21 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 2: .i. CARAC7E?.: S7: C ?? D? SECZTl ZZ (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 2 GTAGCAGGCG GGGATGGGGG G 21 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 3 GTTGTTGGGC ATCTTGCC 18 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 4: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear -fiP (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 4 GTACTTGCGG ATCTAGGC 18 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 5: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 5 GTGGGCCCCG CTCTCGCC 18 [2) INFORMATION FOR SEQUENCE CE II? NT. (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 6 GTCGGGGTAC CTGTCGGC 18 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 7: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) i? (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 7: TGCTTTTCTT TTCTGGGCCT C 21 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 8: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 8 GGCGCCGTGC CGCGTCTTGG TGGCGGCGG 29 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 9: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple '(D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 9: (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 10: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (Cr CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 10 GCCTGTGTCT CTCCTCCT 21 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 11: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear -i? (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: 5EZ. FROM IDENT. NO: 11 GCCTTTCCTG GTTCTCTT 18 (2) INFORMATION FOR ~ IDENTIFICATION SEQUENCE NO: 12: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) - TYPE: nucleic acid (CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 12 GCCTGTGTCT GTCCTCCT 18 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 13 (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NG: 13: TCTCCCTTGG GCTCTGGCTC CTTCTC 26 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 14: (i) CHARACTERISTICS OF THE SEQUENCE: (A) .. LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 14 CTTGCTCCTG GGGGCCTCCT G 21 (2) INFORMATION FOR THE IDENTIFICATION SEQUENCE NO: 15: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY : linear , _ * ..__. • (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 15 CTTGCTCCTG GGGGCCTCCT "G 21 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 16: (i) CHARACTERISTICS OF THE SEQUENCE: (A *) 'LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 16 GGTGTGCGGG GCCTGGTGCC 20 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 17 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA 'ser.Ótrico) (ix) FEATURE: (A) NAME / KEY: misc_feature (B) LOCATION: 12 (D) OTHER INFORMATION: / standard_name = "Reduced A" (ix) FEATURE: (A) NAME / KEY: misc_feature (B) LOCATION: 15 (D) OTHER INFORMATION: / standard_name = "Reduced A" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 17 GGGCGCGGGC GAGCATCGC 19 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 18 (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 18: CCTCCTTCCT GGTCTGTCTG C 21 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 19: (i) CHARACTERISTICS OF THE SEQUENCE: (A). LENGTH: 20 base pairs (7 TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 19 GCCCTGCTGC TCTTTCTGCT 20 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY : linear (ii> TYPE D? MOLECULE: DNA [ger.d.ico] (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 20 GCGCTCGGCC TGGTCCCGG ~ 20 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 21: (i) CHARACTERISTICS OF THE SEQUENCE: k} LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 21 GCGCGGGCCG GGGGCTGCTG GG 22 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 22 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear - (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 22 CCTCTTTTCT GTTTTTCCC 19 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 23: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA. { genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 23 GTTCTTGGCT TCTTCTGTC 19 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 24: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple \ _ ~ * > .? r > iU'J-.h. ._.__..?.AC* (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 24: CTCTGCTGGT TTTCTGCCTT CTGCCC 26 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 16: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 41 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 25 TTTTCTCTTT CGCTTTCTTT TCGTCTCCTG TTCCTCCTTT T 41 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 26: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 33 base pairs (B) TYPE: nucleic acid, C, FCRMA D? THE CADEN; (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 26 CTCTGTCTTG TTCTGGTCCT TCGTGGGGCT CTG 33 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 27: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 27: "CTCTGGTTGG CTTCCTTC 18 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 28: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs' B% TYPE: acci? c n cleicc (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 28 TCCCTGTTTC CCCCCTTT 18 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 29: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 27 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 29: GCTTCTCTTT CGTTCCCGGT GGGCTCG 27 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 30: (i) CHARACTERISTICS OF THE SEQUENCE: (Ai LCNGITTD: 22 pairs? rases (B) TYPE: nucleic acid (C) FORM OF THE CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 30 GCTTGTGTGC? CTGCTGTCT CT 22 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 31: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 39 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 31 TGGTGGGGCT GGGGCTCCGG GGTCTCTGCC CCTCCGTGC 39 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 32: -'i) CHARACTERISTICS D? LA CTENCE: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 32 GTCCTTCTTG TCCGCTGCC 19 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 33 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 33: GTGGGGCCTG CTCTCCCGGC CTCCG 25 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 34: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 34 GGGTCCTCAT GGCTGGGG 18 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 35: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (ix) FEATURE: (A) NAME / KEY: misc_feature (B) LOCATION: 9 (E OTHER INFORMATION: 'standard na-.e = "Eei ce? A" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 35 GGGTCCTCAT GGCTGGGGTC 20 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 36: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 36 GTCTTTGTTT CTGGGCTCGT GCC 23 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 37 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear [i i TYPE OF MOLECULE: DNA (ger.omic) (ix) FEATURE: (A) NAME / KEY: misc_feature (B) LOCATION: 5 (D) OTHER INFORMATION: / standard_name = "Reduced A" (ix) FEATURE: (A) NAME / KEY: misc_feature (B) LOCATION: 14 (D) OTHER INFORMATION: / standard_name = "Reduced A" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 37 GTTCATGGTG GCTAGGTGGG GC 22 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 38: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY : linear (ix) CHARACTERISTICS:] NAME / Cl _-. 77_: my fear re (B) LOCATION: 10 (D) OTHER INFORMATION: / standard_name = "Reduced A" (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 38: GGGGTGGGTA GGCCGTGTCT GGGG 24 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 39: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 39 CGGTTTCCTT TGCGGTC 17 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 40: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: C nucleic acid}. FCEXA D? THE CAD? NA: s ^ -C-Ila (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 40 GTGCTCCGGT GGCTTTTT 18 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 41: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (ix) CHARACTERISTIC: (A) NAME / KEY: misc_feature (B) LOCATION: 6 (D) OTHER INFORMATION: / standard_name = "Reduced A" (ix) CHARACTERISTICS: (A) NAME / KEY: tnisc_f ature (B) "= ICACIIN: 10 (D) OTHER INFORMATION: / stanaard_name =" Re? uce? TO" (xi) DESCRIPTION "OF THE SEQUENCE: ID SECTION NO: 41 GGTCCAGCCA TGGGTCTGGG 20 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 42: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 42 GCTGGTCCTC TGCTGTCCTT GCTG 24 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 43: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 43 GCCCCGTCTG CTGCTCCTCG TGCCG 25 (2) INFORMATION FOR IDENTIFICATION SEQUENCE NO: 44: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) CHAIN FORM: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (ix) FEATURE: (A) NAME / KEY: misc_feature (B) LOCATION: 6 (D) OTHER INFORMATION: / standard_name = "Reduced A" (ix) FEATURE: (A) NAME / KEY: misc_feature (B) LOCATION: 17 (D) OTHER INFORMATION: /sta:_?ard_..a-e= 7 ... ';: e A " (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: 44 GTTTCATCTT GGCTTTATCC 20

Claims (20)

1. A method for treating respiratory tract disease in a subject in need of such treatment, characterized in that it comprises: topically administering an antisense oligonucleotide to the airway epithelium of the subject in an effective amount to treat the disease; and the antisense oligonucleotide that is essentially free of adenosine.

2. The method according to claim 1, characterized in that the respiratory tract disease is a disease of the lung and the epithelium of the respiratory tract is an epithelium of the airways of the lung.

3. The method according to claim 1, characterized in that the antisense oligonucleotide comprises nucleotides in which at least one phosphodiester bond is replaced with a bond selected from the group consisting of methylphosphonate bonds, phosphotriester bonds, phosphorothiotate bonds, linkages of phosphorodithioate and phosphoramidate bonds.

4. The method according to claim 1, characterized in that the respiratory tract disease is selected from the group consisting of systolic fibrosis, asthma, chronic obstructive pulmonary disease, bronchitis, and other diseases of the respiratory tract, characterized by an inflammatory response. .

5. ? rr.? ttdc, of ccr.crcrity ccr. the vindication. 1, characterized in that the antisense oligonucleotide is labeled against an mRNA encoding a protein selected from the group consisting of the human adenosine A2a receptor, the human adenosine A2b receptor, human ß IgE receptor, CD23 antigen of the human Fc-epsilon receptor, histidine human decarboxylase, human beta-tryptase, human tryptase-I, human prostaglandin D synthase, human cyclooxygenase-2, human eosinophilic cationic protenon, human eosinophil-derived neurotoxin, human eosinophil peroxidase, human intercellular adhesion molecule-1 (ICAM-1), human vascular cell adhesion molecule 1 (VCAM-1), human endothelial leukocyte adhesion molecule (ELAM-1), human selectin P, human endothelial monocyte activation factor, human IL-3, human IL-4, IL- Human, human IL-6, human IL-8, neutrophil chemotactic factor derived from human monocyte, neutrophil human elaptase, human neutrophil oxidase factor, cathepsin G huma na, human defensin 1, human defensin 3, human macrophage inflammatory protein-1-alpha, human muscarinic acetylcholine HM1 receptor, human muscarinic acetylcholine HM3 receptor, human fibronectin, human GM-CSF, human tumor necrosis factor a, leukotriene C4 human synthase, human major basic protein and endothelin 1.

6. ? l rr z- z, from or.forri ad ccr. claim 1, characterized in that the antisense oligonucleotide is delivered by administering an aerosol of respirable particles containing the oligonucleotide aptisentide to the lungs of the subject.

7. The method, according to claim 6, characterized in that the particles are selected from the group "consisting of solid particles and liquid particles.

8. The method, according to claim 6, characterized in that the aerosol is comprised of particles having a particle size within the range of about 0.5 to 10 microns.

9. The method according to claim 8, characterized in that the particles are liposomes containing the antisense oligonucleotide.

10. The method according to claim 6, characterized in that the antisense oligonucleotide is administered in an amount sufficient to achieve the intracellular ccr.t.tra.tra.tra.tra.s. Of the oligo cledtido ar. i = er. gone in the subject of aprcx_ ~. amente 0.1 to 10 / _rr ..

11. A pharmaceutical composition, characterized in that it comprises, together with a pharmaceutically acceptable carrier: an antisense oligonucleotide in an amount effective to treat an airway disease; the antisense oligonucleotide that is essentially free of adenosine.

12. The pharmaceutical composition according to claim 11, characterized in that the respiratory tract disease is a disease of the lung and the epithelium of the respiratory tract is an epithelium of the airways of the lung.

13. The pharmaceutical composition according to claim 11, characterized in that the antisense oligonucleotide comprises nucleotides in which at least one phosphodiester bond is replaced with a bond selected from the group consisting of methylphosphonate bonds, phosphotriester bonds, phosphorothiotate bonds, phlogophythioate bonds and fc = í trarr.idatc bonds.

14. The pharmaceutical composition, according to claim 11", characterized in that the disease of the respiratory tract is sistatic fibrosis.

15. The pharmaceutical composition, according to claim 11, characterized in that the antisense oligonucleotide - is labeled against an mRNA that encodes a protein selected from the group consisting of the human adenosine A2a receptor, the human adenosine A2b receptor, the beta receptor Human IgE, the CD23 antigen of the human Fc-epsilon receptor, human histidine decarboxylase, human beta tryptase, human tryptase-I, human prostaglandin D synthase, human cyclooxygenase-2, human eosinophil cationic protein, human eosinophil-derived neurotoxin, eosinophil peroxidase human, human intercellular adhesion molecule-1 (ICAM-1), human vascular cell adhesion molecule 1 (VCAM-1), human endothelial leukocyte adhesion molecule (ELAM-1), human selectin P, monocyte activation factor human endothelial, human IL-3, human IL-4, human IL-5, human IL-6, human IL-8, neutrophil chemotactic factor derived from human monocyte , neutrophil human elaptase, human neutrophil oxidase factor, human cathepsin G, human defensin 1, human defensin 3, protein-1-alpha ir..lar.atoria rr.acrctago hrr.ana, receptor H _-! _ te tet_. Human muscarinic, human muscarinic HM3 receptor, human muscarinic acetylcholine, human fibronectin, human GM-CSF, human tumor necrosis factor a, human leukotriene C4 synthase and human major basic protein.

16. The pharmaceutical composition according to claim 11, characterized in that the antisense oligonucleotide "is delivered by administering an aerosol of respirable particles containing the antisense oligonucleotide to the lungs of the subject.

17. The pharmaceutical composition according to claim 16, characterized in that the particles are selected from the group consisting of solid particles and liquid particles.

18. The pharmaceutical composition according to claim 16, characterized in that the aerosol is comprised of particles having a particle size in the range of about 0.5 to 10 microns.

19. The pharmaceutical composition, according to claim 16, characterized in that the particles are

20. The pharmaceutical composition according to claim 11, characterized in that the antisense oligonucleotide is administered in an amount sufficient to achieve the intracellular concentrations of the antisense oligonucleotide in the subject from about 0.1 to 10 μ. 2? The pharmaceutical composition according to claim 11, characterized in that the antisense oligonucleotide is conjugated to a molecule capable of cellular uptake.