Antisense compositions targeted to beta1-adrenoceptor-specific mRNA and methods of use

Abstract

Disclosed are antisense oligonucleotide, polynucleotide, and peptide nucleic acid compounds that specifically bind to mammalian mRNA encoding a beta1-adrenoceptor polypeptide and that are useful in the control and / or treatment of cardiac dysfunction, hypertension, hypertrophy, myocardial ischemia, and other cardiovascular diseases in an affected mammal, and preferably, in a human subject. The antisense compounds disclosed herein, and pharmaceutical formulations thereof, provide sustained control of beta1-adrenoceptor expression over prolonged periods, and achieve therapeutic effects from as little as a single dose. Administration of these antisense compositions to approved animal models resulted in a decrease in blood pressure, but no significant change in heart rate. Use of such antisense compositions in the reduction of beta1-adrenoceptor polypeptides in a host cell expressing beta1-adrenoceptor-specific mRNA, and in the preparation of medicaments for treating human and animal diseases, and in particular, hypertension and other cardiac dysfunction is also disclosed.

Description

[0001] The present application is a continuation-in-part of U.S. patent application Ser. No. 09 / 152,717 filed Sep. 14, 1998, and issued Jul. 11, 2000 as U.S. Pat. No. 6,087,343, and PCT International patent application Serial No. PCT / US99 / 21007, filed, Sep. 14, 1999, and published Mar. 23, 2000 as WO 00 / 15783, the entire contents of each of which is specifically incorporated herein by reference in its entirety without disclaimer.1.0 BACKGROUND OF THE INVENTION[0003] 1.1 Field of the Invention[0004] The present invention relates generally to the fields of cardiovascular disease and hypertension. More particularly, it concerns antisense oligonucleotide compositions and methods that are useful for reducing hypertension, cardiac hypertrophy, and myocardial ischemia in animals, particularly in mammals such as humans. Specifically, the invention provides antisense oligonucleotide and polynucleotide compositions capable of binding to .beta..sub.1-adrenoceptor (.beta..sub.1-adrenergic recep...

AI Technical Summary

Benefits of technology

[0018] The present invention overcomes these and other limitations in the prior art by providing antisense nucleic acid compounds and compositions comprising them that specifically inhibit or reduce the expression of an mRNA encoding a .beta..sub.1-adrenoceptor (.beta..sub.1-AR) polypeptide in a host cell expressing the mRNA. More specifically, the subject invention provides antisense oligonucleotides (AS-ONs) and antisense oligo-peptide nucleic acids (AS-PNAs) that can specifically bind to a mammalian .beta..sub.1-AR mRNA, resulting in the reduction or inhibition of translation of the messenger ribonucleic acid (mRNA) into .beta..sub.1-AR polypeptide. Such oligonucleotides and PNA may be readily formulated in pharmaceutically-acceptable vehicles and provided directly to host cells, or administered systemically to mammals that express .beta..sub.1-AR mRNA to reduce the level of .beta..sub.1-AR produced in such cells and affected mammals.

[0019] The invention also provides genetic constructs comprising substantially full-length, antisense polynucleotide (AS-PN) sequences operably linked to a suitable promoter that may be used to transform selected cells to endogenously express "antisense" mRNA sequences that are complementary to the native .beta..sub.1-AR mRNA sequence. When expressed in a suitable host cell, these essentially full length anti-mRNAs specifically bind to the native .beta..sub.1-AR mRNA produced in the same host cell, and effectively reduce the availability of native .beta..sub.1-AR mRNA that can serve as a template for the translation machinery of the host cell to produce .beta..sub.1-AR polypeptide.

[0020] The antisense compounds and the genetic constructs of the present invention may be prepared in a variety of compositions, and may also be formulated in appropriate pharmaceutical vehicles for administration to human or animal subjects to inhibit or significantly reduce the expression of .beta..sub.1-AR-specific mRNA, and / or to inhibit or significantly reduce the translation of .beta..sub.1-AR-specific mRNA into functional .beta..sub.1-AR polypeptide. The antisense compounds of the present invention, and compositions comprising them provide new and useful therapeutics for the treatment, control, and amelioration of symptoms of a variety of cardiovascular disorders including hypertension, ischemia, cardiac hypertrophy, myocardial infarction, cardiac dysfunction, and diseases of the heart, that result from, or are exacerbated by, expression of .beta..sub.1-AR-specific mRNA in the host cells of the affected mammal. Moreover, pharmaceutical compositions comprising one or more of the nucleic acid compounds disclosed herein, provide significant advantages over existing conventional therapies-namely, (1) their reduced side effects, (2) their increased efficacy for prolonged periods of time, (3) their ability to increase patient compliance due to their ability to provide therapeutic effects following as little as a single dose of the composition to affected individuals.

Problems solved by technology

Hypertension is the result of increased arterial resistance to blood flow and left untreated can lead to various pathological consequences.

Unfortunately, the short lasting effects of these drugs often require multiple, even daily doses to be therapeutically effective.

Poor compliance is a major problem with drug regimens and can lead to a hypertensive crisis if the drug is not taken as scheduled.

Second, AS-ODNs do not have direct CNS effects, because of the negligible transport of these highly polar molecules through the blood-brain barrier (Agrawal et al., 1991).

Third, antisense elements tested in different systems produce long-term effects after single treatment (Gyurko et al., 1997).

Current .beta.-blocker drugs, however, have certain disadvantages, including: (1) they have to be taken daily, or twice a day and compliance is a problem; (2) they have central effects, leading to psychological changes that contribute to the problem of patient compliance; and (3) many of the .beta.-blockers now available are not specific for .beta..sub.1-ARs and, therefore, can have untoward side effects.

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