By a News Reporter-Staff News Editor at Life Science Weekly — From Alexandria, Virginia, NewsRx journalists report that a patent by the inventors Tachas, George (Kew, AU); Karras, James G. (San Marcos, CA); Gregory, Susan (San Diego, CA); Crosby, Jeffrey R. (Murrieta, CA); Dobie, Kenneth W. (Del Mar, CA); Bennett, Frank C. (Carlsbad, CA), filed on October 20, 2005, was published online on July 1, 2014 (see also Antisense Therapeutics Ltd.).
The patent’s assignee for patent number 8765700 is Antisense Therapeutics Ltd. (AU).
News editors obtained the following quote from the background information supplied by the inventors: “Inflammation is a localized protective response elicited by tissues in response to injury, infection, or tissue destruction resulting in the destruction of the infectious or injurious agent and isolation of the injured tissue. A typical inflammatory response proceeds as follows: recognition of an antigen as foreign or recognition of tissue damage, synthesis and release of soluble inflammatory mediators, recruitment of inflammatory cells to the site of infection or tissue damage, destruction and removal of the invading organism or damaged tissue, and deactivation of the system once the invading organism or damage has been resolved. In many human diseases with an inflammatory component, the normal, homeostatic mechanisms which attenuate the inflammatory responses are defective, resulting in damage and destruction of normal tissue.
“Cell-cell interactions are involved in the activation of the immune response at each of the stages described above. One of the earliest detectable events in a normal inflammatory response is adhesion of leukocytes to the vascular endothelium, followed by migration of leukocytes out of the vasculature to the site of infection or injury. The adhesion of these leukocytes, or white blood cells, to vascular endothelium is an obligate step in the migration out of the vasculature (Harlan, J. M., Blood 1985, 65, 513-525). This response is mediated by the interaction of adhesion molecules expressed on the cell surface of leukocytes and vascular endothelial cells.
“Very late activating antigen-4 (also called VLA-4, .alpha.4.beta.1 or CD49d/CD29) is an integrin expressed in the surface of lymphocytes, monocytes, macrophages, mast cells, basophils and eosinophils. It is a heterodimeric adhesion receptor which is composed of noncovalently linked .alpha.4 and .beta.1 subunits and serves to mediate leukocyte adhesion to vascular cell adhesion molecule-1 (VCAM-1) which is expressed on cytokine-stimulated endothelial cells. This interaction between VCAM-1 and VLA-4 contributes to leukocyte extravasation in acute and chronic inflammatory conditions including multiple sclerosis (MS), rheumatoid arthritis, asthma, psoriasis and allergy.
“The .alpha.4 integrin subunit can also heterodimerize with a .beta.7 integrin chain to form integrin .alpha.4.beta.7 which is known as a mucosal homing receptor because its primary ligand is the mucosal vascular adhesion molecule MadCAM-1. Integrin .alpha.4.beta.7 identifies a subset of memory T cells with a tropism for the intestinal tract, whereas integrin .alpha.4 .mu.l (VLA-4) is constitutively expressed on most mononuclear leukocytes, but not on circulating neutrophils. The interaction of VCAM-1 with VLA-4 suggests that VLA-4 is a potential therapeutic target for inflammatory diseases, including many respiratory conditions, including, for example, asthma and bronchitis (Kassner, P. D., et al, Adv. Exp. Med. Biol. 1992, 323, 163-170).
“Asthma is an inflammatory disease associated with eosinophil infiltration into the lung. VLA-4 is expressed on eosinophils. Metzger, W. J. (Springer Semin. Immunopathol. 1995, 16, 467-478) used a rabbit model of asthma to demonstrate that both an anti-VLA-4 antibody and a CS-1 peptide could reduce eosinophil infiltration into the lung and reduce the development of asthma.
“While steroids and other antiinflammatory drugs are effective in treating inflammatory diseases and conditions, long-term usage often leads to side effects such as increased risk of infection caused by impairment of phagocytic leukocyte migration and function. There is some concern that inhibition of the function of the .beta.1 integrin chain may be associated with increased susceptibility to infections, as demonstrated by a .beta.1 (also called CD18) monoclonal antibody in rabbits (Foster, C. A., 1996, J. Allergy Clin. Immunol., 98, 270-277). It is believed that selective inhibition of the .alpha.4 chain may be a more desirable approach. Inhibition of the .alpha.4 chain is believed likely to reduce levels of the VLA-4 heterodimer as well as the .alpha.4.beta.7 heterodimer.
“Potential therapeutic interventions targeting VLA-4 include monoclonal antibodies and peptide antagonists. Antibodies specific for VLA-4 have been effective in attenuating allergen-driven airway inflammation and hyperresponsiveness in several experimental models of asthma, including the mouse. Leger, O. J. P. et al. (Human Antibodies, 1997, 8, 3-16) describe a monoclonal antibody against VLA-4 that is in phase III clinical trials for multiple sclerosis. CS-1 peptide antagonists have been described by Jackson, D. Y., et al. (J. Med. Chem. 1997, 40, 3359-3369). Hayashi et al. (Cell Struct. Funct. 1991, 16, 241-249) have used a vector expressing RNA complementary to chicken integrin .beta.1 to reduce integrin .beta.1 expression, resulting in altered cell attachment and shape.
“Antisense oligonucleotides (‘ASOs’) targeted to various integrins have been used as tools to dissect the functional interactions of integrins in complex settings. Lallier and Bronner-Fraser (Science, 1993, 259, 692-695) have used phosphorothioate oligonucleotides targeted to conserved and nonconserved regions of chick .beta.1, human .alpha.4, rat .alpha.1 and human .beta.5 integrins to determine the effects of these integrins on cell attachment. These same oligonucleotides were also injected into cranial neural crest migratory pathways in avian embryos, and it was demonstrated that those oligonucleotides that inhibited cell attachment in vitro also caused neural crest and/or neural tube abnormalities in vivo (Kit et al., Devel. Biol. 1996, 179, 91-101).
“EP patent application 688 784 (Carolus et al.) discloses 3′ derivatized oligonucleotide analogs, including one sequence targeted to the .beta.1 subunit of VLA-4.
“U.S. Pat. Nos. 5,968,826 and 6,258,790 (Bennett et al.) describes modulating integrin .alpha.4 expression through the use of antisense oligonucleotides targeted to nucleic acids encoding integrin .alpha.4. U.S. Pat. No. 5,968,826 discloses that such antisense oligonucleotides can be used to treat a large variety of inflammatory diseases associated with VLA-4 expression, including asthma. According to this document, in general, a dosage of from 0.01 .mu.g to 100 g of antisense oligonucleotide per kg of body weight, which may be given once or more times daily, weekly, monthly or yearly, or even every 2 to 20 years, may be used to treat the inflammatory diseases. The range of dosages exemplified for the various inflammatory diseases is from 0.01 mg to 20 mg of antisense oligonucleotide per kg body weight. Example 30 describes the prophetic use of antisense oligonucleotides in a murine model for asthma in which mice are injected intravenously with 1 mg/kg to 5 mg/kg doses of antisense oligonucleotides to integrin .alpha.4.”
As a supplement to the background information on this patent, NewsRx correspondents also obtained the inventors’ summary information for this patent: “We have now found that respiratory diseases and conditions associated with airway hyperresponsiveness, eosinophilia, neutrophilia, leukocytes or overproduction of mucus and/or with the expression of integrin. .alpha.4 can be successfully treated or prevented using very low doses of antisense compounds targeted to nucleic acids encoding integrin .alpha.4.
“Thus, the invention provides a method for the treatment and/or prophylaxis of an animal having a respiratory disease or condition associated with airway hyperresponsiveness, eosinophilia, neutrophilia, leukocytes or overproduction of mucus and/or with the expression of integrin .alpha.4 comprising administering to the animal a composition comprising from 0.001 to 1000 .mu.g per kg body weight of the animal of an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4.
“The invention further provides a method for the treatment and/or prophylaxis of an animal having a respiratory disease or condition associated with airway hyperresponsiveness, eosinophilia, neutrophilia, leukocytes or overproduction of mucus and/or with the expression of integrin .alpha.4 comprising administering to the animal a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 at a dosage level of from 0.001 to 1000 .mu.g of the antisense compound per kg body weight of the animal.
“The invention further provides for the use of an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 in the manufacture of a medicament for the treatment and/or prophylaxis of an animal having a respiratory disease or condition associated with airway hyperresponsiveness, eosinophilia, neutrophilia, leukocytes or overproduction of mucus and/or with the expression of integrin .alpha.4 in which the medicament is to be administered at a dosage level equating to from 0.001 to 1000 .mu.g of the antisense compound per kg body weight of the animal.
“The invention further provides for a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 for use in therapy, in which the antisense compound is dosed at a level of from 0.001 to 1000 .mu.g of the antisense compound per kg body weight of the animal being treated.
“The use of such low dosages provides significant benefits including significantly reducing the potential for unwanted side-effects and providing a considerable cost saving in terms of the cost of manufacture per unit dose. In addition, it allows for greater flexibility in the potential devices of administration.
“Suitably, the antisense compound is dosed at a level of at least 0.005, preferably at least 0.01, more preferably at least 0.05, more preferably at least 0.1, more preferably at least 0.5, yet more preferably at least 1 and yet more preferably at least 2 .mu.g per kg body weight of the individual animal. The antisense compound may be dosed a higher levels such as, for example, at least 5 .mu.g per kg body weight of the individual animal.
“Suitably, the antisense compound is dosed at a level of less than 1000, preferably less than 500, more preferably less than 200, more preferably less than 150, more preferably less than 100, more preferably less than 75, more preferably less than 50, more preferably less than 20 and yet more preferably less than 10 .mu.g per kg body weight of the individual animal. The antisense compound may be dosed at lower levels such as, for example, less than 7, less than 5, less than 2 or less than 1 .mu.g per kg body weight of the individual animal.
“Suitably, the antisense compound is dosed at a level of no more than 1000, preferably no more than 500, more preferably no more than 200, more preferably no more than 150, more preferably no more than 100, more preferably no more than 75, more preferably no more than 50, more preferably no more than 20 and yet more preferably no more than 10 .mu.g per kg body weight of the individual animal. The antisense compound may be dosed at lower levels such as, for example, no more than 7, no more than 5, no more than 2 or no more than 1 .mu.g per kg body weight of the individual animal.
“Furthermore, we have found that antisense compounds targeted to nucleic acids encoding integrin .alpha.4 are especially effective when delivered topically. The finding that topical administration is particularly effective is surprising given the pharmacokinetics of antisense and/or the predominant mechanism of action of integrin .alpha.4 to modulate adhesion and transmigration of white blood cells from the blood into organs.
“Thus, the invention further provides a method for the treatment and/or prophylaxis of an animal having a respiratory disease or condition associated with airway hyperresponsiveness, eosinophilia, neutrophilia, leukocytes or overproduction of mucus and/or with the expression of integrin .alpha.4 comprising topically administering to the animal a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4.
“The invention further provides for the use of an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 in the manufacture of a medicament for the treatment and/or prophylaxis of an animal having a respiratory disease or condition associated with airway hyperresponsiveness, eosinophilia, neutrophilia, leukocytes or overproduction of mucus and/or with the expression of integrin .alpha.4 in which the medicament is to be administered topically.
“The invention further provides for a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 for use in therapy or prophylaxis, in which the antisense compound is to be administered topically.
“Any suitable method of topical administration to the respiratory system or airway may be used including via the mouth or nose. Topical administration may be to any part of the respiratory system comprising the nose, throat, larynx, trachea, bronchial tubes and the lungs or to airways including the mouth and sinuses. Inhalation or insufflation are particularly preferred. Preferred routes are pulmonary, intranasal and intratrachael administration.
“Thus, the invention also provides a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 in a formulation suitable for inhalation or insufflation.
“Furthermore, the invention provides a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 in a formulation suitable for intranasal, intrapulmonary or intratracheal administration.
“Preferably the composition containing the antisense compound is powdered or aerosolised and inhaled by the individual. Suitably, the composition is administered through a metered dose inhaler (MDI), nebuliser, dry powder inhaler (DPI), nasal inhaler or as nasal drops. This offers significant advantages in terms of ease and simplicity of use. Choice of device also enables delivery to different parts of the respiratory system. Nasal drops or nasal inhalers are often used to deliver to the upper respiratory tract such as the nose for treatment of nasal conditions such as rhinitis, whereas MDI and DPI are often used to deliver to the lower respiratory tract for treatment of conditions such as asthma.
“The ability of low doses of oligonucleotides to work when administered topically suggests that there is one or more mechanisms being effected at a predominantly topical, local level. Although not wanting to be bound by mechanism, it is proposed that the predominant mechanism of action of integrin .alpha.4 is to modulate adhesion and transmigration of white blood cells from the blood into organs. An intravenous, intraperitoneal, or subcutaneous route for delivery of the antisense drug to integrin .alpha.4 to white blood cells will predominantly interfere with the adhesion of .alpha.4 positive white blood cells to vascular endothelium to thereby interfere with the obligate step of migration of the integrin .alpha.4 positive white blood cells out of the vasculature to the respiratory system, the lung being the site of asthma. The use of a pulmonary route provides significant benefits including improved specificity for the integrin .alpha.4 expressed in lung or significant reduction of the potential for unwanted systemic side-effects involved in modulating integrin .alpha.4 in white blood cells which are involved in the normal surveillance of organs other than the lung such as the brain, knee, and skin. Clearly, the ability to achieve a local effect is highly beneficial, systemic responses having the potential to produce unwanted side-effects.
“Previous integrin .alpha.4 inhibitors described in the prior art have typically relied on dosing regimes requiring more than one dose being administered daily. In addition, they have typically relied on dosage units considerably higher than those of the present invention. For example, Koo G. C. et al. (Am. J. Respiratory Crit. Care Med, Vol. 167, pp 1400-1409, 2003) dosed twice daily with a topical composition and reported that occupancy was good when dosing at 1 and 3 mg/kg.
“We have found that the present invention can provide effective treatment of respiratory diseases or conditions when dosed once a day and even once every two days. This is significant because research indicates that there is a poor compliance rate (about 30-70\%) in the case of drugs, such as corticosteroids, which have to be inhaled twice or more (maybe up to five times) daily.
“Thus, the invention further provides a method for the treatment and/or prophylaxis of an animal having a respiratory disease or condition associated with airway hyperresponsiveness, eosinophilia, neutrophilia, leukocytes or overproduction of mucus and/or with the expression of integrin .alpha.4 comprising administering to the animal a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 no more than once daily.
“The invention further provides for the use of an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 in the manufacture of a medicament for the treatment and/or prophylaxis of an animal having a respiratory disease or condition associated with airway hyperresponsiveness, eosinophilia, neutrophilia, leukocytes or overproduction of mucus and/or with the expression of integrin .alpha.4 in which the medicament is to be administered no more than once daily.
“The invention further provides a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 for use in therapy or prophylaxis, in which the antisense compound is to be administered no more than once daily.
“The invention further provides a kit comprising a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 and a device which allows the composition to be administered by inhalation or insufflation at a dosage level of from 0.001 to 1000 .mu.g of the antisense compound per kg body weight of the animal.
“The invention further provides a kit comprising a composition comprising an antisense compound targeted to a nucleic acid molecule encoding integrin .alpha.4 and instructions that the composition is to be administered at a dosage range of from 0.001 to 1000 .mu.g of the antisense compound per kg body weight of the animal being treated, optionally with instructions that the composition is to be administered by inhalation or insufflation and/or no more than once a day.
“The present invention also provides antisense compounds, particularly oligunucleotides, which are targeted to a nucleic acid encoding integrin .alpha.4. Thus, the invention further provides antisense compounds targeted to a nucleic acid molecule encoding integrin .alpha.4, wherein the antisense, compound is an antisense oligonucleotide comprising at least an 8 nucleobase portion of one of SEQ ID Nos 103 to 178. Preferably, the antisense oligonucleotide comprises at least a 10, more preferably at least a 13 and yet more preferably at least a 15 nucleobase portion from any one of SEQ ID Nos 103 to 178.
“Without being limited by theory, it is believed that VLA-4 (.alpha.4.beta.1) is involved in several pathophysiological processes underlying the disease asthma and other respiratory system conditions and .alpha.4.beta.7 may also have a role in these conditions. The predominant mechanism of action of integrin .alpha.4 is to modulate adhesion and transmigration of white blood cells i.e leukocytes from the blood into organs such as the lung. The VLA-4 binds to VCAM on cytokine-stimulated endothelial cells which is important in the transmigration of the white blood cells, particularly eosinophils into lung and nasal passages. Once in the lung or nasal passage, the role of integrin .alpha.4 is less clear. VLA-4 activation and/or .alpha.4.beta.7 may contribute to local inflammatory processes, bronchoconstriction, and mucus production, exacerbating symptoms by a number of potential mechanisms (see FIG. 10). Mast cells (.alpha.4.beta.1/.alpha.4.beta.1) and basophils (.alpha.4.beta.1) release agents involved in bronchoconstriction and neutrophil activation and eosinophil activation (.alpha.4.beta.1/.alpha.4.beta.7) which lead to airway inflammation. Macrophages (.alpha.4 .mu.l), B-cells (.alpha.4 .mu.l), and T cells including Th2 positive leukocytes (.alpha.4.beta.1/.alpha.4.beta.7) together with eosinophils may also be involved in the airway inflammation as are airway epithelial cells (.alpha.4.beta.1). Leukocytes are also believed to attach to local smooth muscle cell in the lung via VLA-4 which may contribute to bronchospasm. Finally, VLA-4 is involved in neovascularization and angiogenesis is important in lung remodelling which occurs in several diseases of the lung and nasal passage.
“The methods and compositions of the present invention may be used to treat any respiratory disease or condition associated with airway hyperresponsiveness, eosinophilia, neutrophilia, leukocytes or overproduction of mucus and/or with the expression of integrin .alpha.4. Such diseases or conditions will be evident to the skilled person. Examples of respiratory conditions or diseases already know to be associated with an overproduction of mucus with can be treated by the methods and compositions of the present invention include, for example, chronic respiratory conditions like asthma, cystic fibrosis, alpha-1 antitrypsin deficiency, chronic obstructive pulmonary disease and chronic bronchitis. It will be understood that reference to respiratory herein includes the nose, throat, larynx, trachea, bronchial tubes and the lungs or to the passages filled with air such airways including the mouth and sinuses. and that the compositions of the invention can be used to treat diseases and conditions associated with any the aforementioned. For example, the compositions are potentially useful in treating conditions like rhinitis, where some of the effects of the disease manifest themselves in the upper respiratory tract, and in treating sinusitis and in treating diseases or conditions associated with leukocytes, neutrophils, eosinophilia or dependent on airway hyperresponsiveness (AHR).
“It will be understood that, in relation to compositions, compounds, components, ingredients or the like described herein, any lower range limit described in relation to a particular composition, compound, component, ingredient or the like may be combined with any upper range limit described in relation to the same composition, compound, component, ingredient or the like to define a suitable range for that particular composition, compound, component, ingredient or the like.
“Throughout this specification the word ‘comprise’, or variations such as ‘comprises’ or ‘comprising’, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.
“Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.”
For additional information on this patent, see: Tachas, George; Karras, James G.; Gregory, Susan; Crosby, Jeffrey R.; Dobie, Kenneth W.; Bennett, Frank C.. Topical Administrations of Antisense Compounds to VLA-4 for the Treatment of Respiratory Conditions. U.S. Patent Number 8765700, filed October 20, 2005, and published online on July 1, 2014. Patent URL: http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO1&Sect2=HITOFF&d=PALL&p=1&u=\%2Fnetahtml\%2FPTO\%2Fsrchnum.htm&r=1&f=G&l=50&s1=8765700.PN.&OS=PN/8765700RS=PN/8765700
Keywords for this news article include: Antibodies, Antisense Therapeutics Ltd., Immunology, Phagocytes, Blood Cells, Eosinophils, Neutrophils, Granulocytes, Blood Proteins, Immunoglobulins, Endothelial Cells.
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