WO1999037761A1 - Novel sequence variants of the human beta2-adrenergic receptor gene and use thereof - Google Patents

Abstract

The invention relates to novel sequence variants of the human beta2-adrenergic receptor gene and to their use for diagnosing a range of diseases, especially for detecting a predisposition to high blood pressure, for diagnosing reactivity to therapeutic agents where this varies from one case to the next, and for developing therapeutic agents on the basis of pharmacogenetic principles.

Description

New sequence variants of the human beta2-adrenergic receptor gene and their use

The invention relates to new sequence variants of the human beta2-adrenergic receptor gene and their use for the diagnosis of a spectrum of diseases, in particular for the determination of hypertension dispositions and for the development of therapeutic agents on the basis of pharmacogenetic principles.

The human beta2-adrenergic receptor is an important component of the sympathetic nervous system and, as such, regulates a spectrum of central and peripheral functions, e.g. Cardiovascular functions, metabolic functions, central nervous functions and neurosecretion. It is the target of pharmaceuticals / therapeutic agents with a wide range of indications, which are among the most commonly prescribed drugs. A variety of findings indicate that this receptor could play a role in the pathogenesis / pathophysiology of a number of common diseases, e.g. hypertension and other cardiovascular diseases, various neuropsychiatric diseases such as Depression, and metabolic diseases such as Obesity (Insel PA (Ed) (1987) Adrenergic receptors in man, Marcel Dekker, New York, Basel).

The aim of the invention is to determine variants, polymorphisms, mutations and resulting haplotypes in the DNA sequence of the human beta2-adrenergic receptor gene and to determine their correlations with disease dispositions. Based on these correlations, a method for diagnosing these disease predispositions, for predicting severity, course and survival time, a system for predicting individual responsiveness to beta2 active therapeutic agents, for developing individually specific beta2 receptor agonists and antagonists, and a system for developing a new one Class of beta2 effective therapeutic agents, as well as the development of test systems for researching pathophysiological relationships and development of the above-mentioned therapeutic agents. In summary, an individually optimal therapeutic agent can be predicted or developed for each beta2 genotype. The object is achieved according to the claims, the subclaims are preferred variants. 2

It was found that, in addition to the 3 mutations already known in the coding region (at positions 1633, 1666 and 2078), further variants are present in the 5 'regulating region of the sequence of the human beta2-adrenergic receptor gene. It has also been found that these genetic variants are predisposed to various diseases, e.g. B. hypertension correlate.

The invention then relates to the sequence of the human beta2-adrenergic receptor gene, which is at positions 159, 245, 565, 934, 1120, 1221, 1541, 1568, 1633, 1666, 1839, 2078, 2110, 2640, and 2826 in whole or is partially mutated. It is in particular a sequence which completely or partially the mutations T- »A (position 159), A → G (position 245), G → A (position 565), G → A (position 934), G → C (Position 1120), C → T (position 1221), C → T (Arg-> Cys) (position 1541), T → C (position 1568), A → G (Arg-> Gly) (position 1633), C → G (Gln → Glu) (position 1666), G-> A (position 1839), C-> T (Thr-> Ile) (position 2078), C-> A (position 2110), G-> C ( Item 2640), and G-> A (Item 2826) contains (Figures 1, 2a and 2b).

The following sequences (haplotypes) are particularly important:

Sequence with mutations 1541 T, 1633 A and 1666 C,

Sequence with mutations 1541 C, 1633 G and 1666 G,

Sequence with mutations 1541 T, 1633 G and 1666 C,

Sequence with mutations 1541 T, 1568 T, 1633 A and 1666 C,

- Sequence with the mutations 1541 C, 1568 C, 1633 G and 1666 G as well as the

- Sequence with mutations 1541 T, 1568 T, 1633 G and 1666 C.

The invention furthermore relates to a method for determining disease dispositions, it being possible for all sequences and variants of the beta2-adrenergic receptor gene from the individual mutation to all possible combinations of all variants (including any absolute number of variants which can be included) to be genotyped and enable the corresponding statements about disease dispositions.

The method is characterized in that the DNA of a test subject is isolated and genotyped at least in one of the exchanged positions and subsequently compared with the reference DNA sequence. Embodiments are preferred in which at least position 1633, at least the three positions 1541, 1633 and 1666 or the four last-mentioned positions (1541, 1568, 1633 and 1666) or at the seven positions 245, 565 934, 11541, 1568, 1633 and genotyped in 1666. 3

The method can also be varied by genotyping at least 3 of the 4 positions 1541, 1568, 1633 and 1666 and subsequently comparing them with the reference DNA sequence. Genotyping of positions 1541, 1633 and 1666 is preferred here.

Genotyping is performed by sequencing or by other methods that are suitable for the detection of point mutations. This includes PCR-based genotyping methods such as B. allele-specific PCR, other genotyping methods using ohgonucleotides (examples would be 'dot blotting' or 'oligonucleotide ligation assays' (OLA)), methods using restriction enzymes, and' single nucleotide polymorphism '(SNP) analysis using' matrix assisted laser desorption / ionization mass spectrometry (MALDI), as well as in principle any future method for variant detection including chip technology in all its technological versions.

Proceeding from this, the method according to the invention is suitable for determining a broad spectrum of the most diverse disease dispositions.

In one embodiment, e.g. to determine a disposition for high blood pressure (or the prediction of the range of individual blood pressure values as such), and other cardiovascular diseases, including myocardial infarction and stroke, in the broadest sense the development of end-stage renal failure (requiring dialysis).

Another preferred embodiment variant allows e.g. the determination of a disposition for neuropsychiatric diseases such as depression and anxiety disorders, attention deficit disorder (with hyperactivity), eating disorders, e.g. for anorexia nervosa and bulimia, or disorders caused by post-traumatic stress; or for diseases of the autonomic nervous system, e.g. Bradbury-Eggleston, Sky-Drager and Riley-Day syndrome as well as selective noradrenergic and baroreceptor dispositions, or migraines.

It is also suitable for the detection of dispositions for allergic diseases, especially astiima and atopic disorders.

Another area of application is the determination of a disposition for metabolic diseases such as obesity (as well as familial 'morbid obesity'), including a prediction of the weight range as such or a disposition for weight change, and finally a prediction of the ratio of the body measurements as such, as they are e.g. Express 'body mass index' (BMI). 4

Furthermore, the method also allows the course and severity of diseases to be determined, and the prediction of survival after serious medical diseases, e.g. after myocardial infarction, heart failure and / or stroke.

A further preferred embodiment variant enables the determination of an individually different reactivity of the autonomic nervous system, in particular on endogenous and exogenous stress (as expressed, for example, in particular by an individually different disposition to changes in blood pressure and / or heart rate (deflections) on endogenous and exogenous stress comes), or by individually different blood pressure changes to endogenously or exogenously induced changes in the salt concentration in the blood (individually different salt sensitivity or resistance), and in the broadest sense also by individually different salt and water regulation or reabsorption in the kidney (related volume regulation ) is expressed.

Another important object of the invention is the use of the claimed sequence variants a) for predicting the individually different responsiveness to previously known therapeutic agents (beta2 receptor ligands) and the individually different responsiveness to the endogenous ligands adrenaline and noradrenaline; b) preferably for the development of individually specific beta2 receptor agonists and antagonists; c) in particular also for the development of a new class of therapeutic agents which are directed to the beta2 receptor gene at the 5 'regulatory region, promoter region, in particular e.g. attack the leader peptide and act by regulating transcription, translation and by influencing their efficiency, primarily by regulating expression.

In this context, another object of the invention is the prediction of individual habituation to drug administration (tachyphylaxis), as well as a different disposition for drug side effects. Overall, it is possible to predict individually optimal therapeutic agents, which are based on different mechanisms of action.

Another important object of the invention is the use of the claimed sequence variants for the construction of genes or vectors, in particular for the development of pharmaceutically relevant substances and for the development of a diagnostic kit or any diagnostic method. Such kits or methods can advantageously be used to predict individual disease disposition or individual responsiveness to various beta2 therapeutics. 5

Cultures (cells) that express the various combinations of individual ß2 variants mentioned can thus serve as test models for the development of individually specific therapeutic agents (ß2 agonists and antagonists, as well as beta2 expression-regulating DNA therapeutic agents). This corresponds to test models in vitro, but also in vivo test models are included (transgenic animals that carry these individual receptor variants).

As individual test models, they allow in vitro (= ex vivo) prediction of the individual functional state of the beta2 receptor or the functions mediated by it.

The scope of the claimed invention is detailed below. To develop the invention, the entire known DNA sequence of the human beta2-adrenergic receptor gene, including its regulating and coding regions, was examined in patients and controls by means of the 'multiplex PCR sequencing', and a number of genetic variants were initially identified. Eight new variants were discovered in the 5 'regulating region, the most important of which appears to be the exchange of a highly conserved Arg-> Cys in the' leader peptide 'of the gene (position 1541), which regulates the translation of the receptor gene (position -47 relative to the translation start point), ie its expression.

Summary of the newly identified variants (nucleotide position before the exchange is in relation to the published beta2 receptor sequence, (Kobilka BK et al, Proc.Natl.Acad.Sci USA; 84 (1): 46-50 (1987) [Acc. No . J02960]; the information in brackets after the exchange refers to the translation start):

159 T → A (-1429)

245 A → G (-1343)

565 G → A (-1023)

934 G → A (-654)

1120 G → C (-468)

1221 C → T (-367)

1541 C → T (-47) Arg → Cys exchange in the leader peptide of the beta2-receptor gene

1568 T → C (-20)

These variants are clearly shown in Figures 1, 2a and 2b. 6

Correlations with diseases or clinically relevant phenotypes: Specific influences of the two previously known mutations Arg-> Gly (at position +46 relative to the translation start point, corresponds to position 16 of the amino acid sequence) and Gln-> Glu (at position +79 relative to the translation start point, corresponds Position 27 of the amino acid sequence), as well as the newly discovered 'Leader Peptide' mutation Arg-> Cys (at position -47 relative to the translation start point) on a number of clinically and pathogenetically relevant phenotypes have been demonstrated in several studies. A significant association of the alleles at position 16 of the amino acid sequence with a genetic predisposition to hypertension and extreme deflected blood pressure values was found. Furthermore, the three mutations described have a significant effect on phenotypic parameters such as heart rate, noradrenaline concentrations, blood pressure changes on experimentally induced physical and mental stress, 'coping styles' and personality dimensions, as well as weight and weight change. In particular, an association of the leader peptide mutation with hypertension was also shown. Furthermore, a relationship between beta2-agonist-induced vasodilation and beta2 receptor mutations, preferably at position 16 of the amino acid sequence, and a relationship between beta2-receptor expression in fibroblast cultures of genotyped individuals and beta2 receptor mutations, preferably at position 16 of the amino acid sequence, could be established.

Detection of specific combinations of three of the mutations at positions (relative to the published beta2 sequence, Kobilka et al. 1987) 1541 C → T ('Leader Peptide' mutation Arg-> Cys), 1633 A -> G (Arg-> Gly), and 1666 C -> G (Gin-> Glu): combination 1: 1541 T (Cys allele), 1633 A (Arg allele), 1666 C (Gin allele) combination 2: 1541 C (Arg allele), 1633 G (Gly allele ), 1666 G (Glu Allel) combination 3: 1541 T (Cys Allel), 1633 G (Gly Allel), 1666 C (Gin Allel)

These three specific combinations occur in 80 - 95% of the population, they appear to be evolutionarily selected from the total number of combinations to be expected and represent different functional states of the human beta2-adrenergic receptor which underlie the variability of physiological and pathophysiological functions. In particular, they are associated with an individually different responsiveness to endogenous ligands such as adrenaline and norepinephrine, as well as a different therapeutic responsiveness to beta2 receptor agonists and antagonists, which can make these 'combinations' the starting point for the development of 'individually tailored pharmacotherapy'. 7

Detection of specific beta2- 'haplotypes' consisting of four variants: at positions (relative to the published beta2 sequence, Kobilka et al. 1987) 1541 C → T ('Leader Peptide' mutation Arg-> Cys), 1568 T-> C; 1633 A -> G (Arg-> Gly), and 1666 C -> G (Gln-> Glu):

Combination 1: 1541 T (Cys Allel), 1568 T, 1633 A (Arg Allel), 1666 C (Gin Allel) Combination 2: 1541 C (Arg Allel), 1568 C, 1633 G (Gly Allel), 1666 G (Glu Allele) combination 3: 1541 T (Cys Allele), 1568 T, 1633 G (Gly Allele), 1666 C (Gin Allele)

Combination 1 was observed significantly more frequently in individuals who were inherited with hypertension and is therefore a genetic risk factor.

Detection of specific beta2 'haplotypes' consisting of seven variants: Taking all variants into account, 'haplotypes' consisting of seven variants (including the three mutations mentioned) could be extracted; The calculations were based on the goal of identifying 'haplotypes' from the entirety of the genome which were sufficient to distinguish the patient group from the control group. A specific 'haplotype', combination 1, can be observed more frequently in cases of genetic hypertension, and this can be extended to other phenotypes.

Combination 1: 245 G, 565 G, 934 A, 1541 T (Cys allele), 1568 T, 1633 A (Arg allele),

1666 C (gin allele)

Combination 2: 245 A, 565 A, 934 G, 1541 C (Arg allele), 1568 C, 1633 G (Gly allele),

1666 G (Glu allele)

Combination 3: 245 G, 565 G, 934 G, 1541 T (Cys Allel), 1568 T, 1633 G (Gly Allel),

1666 C (gin allele)

These last described 'haplotypes' finally describe the real, overall individual functional state of the receptor. The invention is based on the concept that it is not individual mutations which are based on different functional (dysfunctional) receptor states, but these are caused by the individual 'polymorphic' overall gene sequence as a function-determining unit. 8th

The invention is explained in more detail by an embodiment.

material and methods

The multiplex PCR sequencing method was used for the collection of the entire polymorphic spectrum of the beta2-receptor gene. For this purpose, the entire promoter region known to date and the coding region were divided into eight fragments and amplified by means of PCR (see FIG. 1). These PCR fragments were pooled and sequenced simultaneously. The fragments of the terrination reactions were separated on a sequence gel and transferred to a nylon membrane by direct transfer electrophoresis (DTE). The individual sequence ladders were decoded by successive hybridization with specific oligonucleotides. The specific conditions for the amplification were as follows: For fragment I, the forward primer ADRBR-F1 with the sequence 5 - TATTGGCCAGGATCTTTTGCTTTCTAT-3 ^' and the reverse primer ADRBR-R1 with the sequence 5 ^' -TAACATTAAGAACATTTTGAAGC-3 ^'were used. Fragment II was amplified with hooves of the two primers ADRBR-F2: 5 ^' -GCATACCCCCGCTCCAGATAAA-3 ^' and ADRBR-R2: 5 ^' -GCACGCACATACAGGCACAAATAC-3 ^' . For fragment III it was the two primers ADRBR-F3: 5 ^' -GGCCGCGTTTCTGTGTTGG-3 ^' and ADRBR-R3: 5 ^' -AGTGCGTTCTGCCCGTTATGTG-3 ^' . For fragment VIII the two primers ADRBR-F8: 5 ^' -GGTACTGTGCCTAGCGATAAC-3 ^' and ADRBR-R8: 5 ^' - TAAAATACCCCGTGTGAGCAAATAAGAG-3 ^' . The reaction conditions for these four fragments were as follows: 10 x PCR buffer (100 mM Tris-HCl, 15 mM MgCl ₂ x 6 H ₂ O, 500 mM KC1, pH 8.3), dNTP 2 mM, 30 μM Primer F, 30 μM primer R, 50 ng genomic DNA and 5 U of a Taq DNA polymerase. Three fragments were amplified with the following temperature profile: 94 ^° C for 4 min; 35 cycles: 94 ^° C 30 sec, 60 ^° C 30 sec, 72 ^° C 1 min and finally 72 ^° C 10 min.

Fragment IV was generated using the two primers ADRBR-F4: 5 - GGGGAGGGAAAGGGGAGGAG-3 ^' and ADRBR-R4: 5 ^' -

CTGCCAGGCCCATGACCAGAT-3 ^' amplified. For fragment VII, the primers ADRBR-F7: 5 ^' -CTGGCTGCCCTTCTTCATCGTT-3 ^' and ADRBR-R7: 5 ^' -

TACCCTCAAGTTAAATAGTCTGTT-3 ^' used. The conditions for these two PCR reactions were as follows: 10 x PCR buffer (160 mM (NH ₄ ) ₂ SO ₄ , 0.1% Tween-20, 500 mM KOH, pH), dNTP 2 mM, 30 μM Primer F , 30 μM Primer R, 50 ng genomic DNA and 4 U of a mixture of the Taq DNA polymerase and a thermostable inorganic pyrophosphatase from Thermits thermophilus. Both fragments were amplified with the following temperature profile: 94 ^° C for 4 min; 35 cycles: 94 ^° C 30 sec, 66 ^° C [Fragment IV] or 60 ^° C [Fragment VII] 30 sec, 72 ^° C 1 min and finally 9

at 72 ^° C for 10 min.

Fragment V was amplified using the two primers ADRBR-F5: 5 - ATGCGCCGGACCACGAC-3 ^' and ADRBR-R5: 5 - GTAGAAGGACACGATGGA-3, fragment VI using the two primers ADRBR-F6: 5 - GCTACTTTGCCATTACTTCACC-3 ^' and ADRBR-R6: 5 ^' -

AAATCTGGGCTCCGGCAGTAGATAAG-3 ^' . These two fragments were amplified using Perkin Elmer's 'AmpliTaq Gold Kit'. The temperature profile for these two fragments was as follows: 94 C 10 min; 35 cycles: 94 C 30 sec, 56 ^° C [fragment V] or 58 ^° C [fragment VI] 30 sec, 72 ^° C 1 min and finally 72 ^° C 10 min.

The sequencing was done using the 'Thermo Sequenase cycle sequencing kit' from Amersham. The PCR primers described above were used as sequencing primers. The sequencing was carried out in four multiplex pools. Pool 1 contained the sequencing primers ADRBR-Fl, ADRBR-F3, ADRBR-F5 and ADRBR-F7; Pool 2 the sequencing primers ADRBR-Rl, ADRBR-R3, ADRBR-R5 and ADRBR-R7. The PCR fragments I, III, V and VII were used in both sequencing pools. Pool 3, on the other hand, contained the sequencing primers ADRBR-F2, F4, F6 and F8; Pool 4 the sequencing primers ADRBR-R2, R4, R6 and R8. The fragments π, IV, VI and VIII were inserted into these two pools.

All PCR and sequencing reactions were carried out in a PTC 225 cycler from MJ Research.

The products of the sequence reactions were separated on a 100 μm thick acrylamide gel (5% acrylamide, 7 M urea) and transferred to a Biodyne A membrane (Pall) under standard DTE conditions (see Richterich and Church, 1993). The membrane was then hybridized with ³² P-labeled ougonucleotides and the individual sequence conductors were detected using a phosphofluorimager (Storm 860, Molecular Dynamics).

10

Literature:

Kobilka BK, Dixon RA, FrieUe T., Dohlman HG, Bolanowski MA, Sigal IS, Yang Feng TL, Francke U., Caron MG, Lefkowitz RJ: cDNA for the beta 2-adrenergic receptor: a protein with multiple membrane-spanning domains and encoded by a gene whose chromosomal location is shared with that of the receptor for platelet-derived growth facto. Proc NatlAcad Sei USA .; 84 (1): 46-50 (1987).

Parola AL and KobUa BK The peptide product of a 5 ^' leader cistron in the beta 2 adrenergic receptor mRNA inhibits receptor synthesis. J Biol Chem. 269 (6): 4497-505 (1994).

Richterich P. and Church G.M .: DNA sequencing with direct transfer electrophoresis and nonradioactive detection. Methods Enzyrnσl. 218: 187-222 (1993).

Legends for the pictures:

illustration 1

Polymorphic spectrum of the human beta 2-adrenergic receptor gene variants are shown according to their nucleotide position (reference sequence Kobilka et al. 1987).

Figure 2 a

Sequence of the human beta 2-adrenergic receptor (Kobilka et al. 1987) variants are shown according to their position.

Figure 2 b

Sequence of the human beta 2-adrenergic receptor (Kobilka et al. 1997). The variants (nucleotide or amino acid exchange) are shown.

CORRECTED SHEET (RULE 91) ISA / EP

Claims

11 Patent claims 1. Sequence of the human beta2-adrenergic receptor gene, characterized in that the bases at positions 159, 245, 565, 934, 1120, 1221, 1541, 1568, 1633, 1666, 1839, 2078, 2110, 2640, and 2826 entirely or partially replaced. 2. Sequence according to claim 1, characterized in that it completely or partially the base exchanges T → A (position 159), A → G (position 245), G → A (position 565), G → A (position 934), G → C (position 1120), C → T (position 1221), C → T (position 1541), T → C (position 1568), A → G (position 1633), C → G (position 1666), G → A (Position 1839), C → T (position 2078), C → A (position 2110), G → C (position 2640), and G → A (position 2826). 3. Sequence according to claim 1 and 2, characterized by the mutations 1541 T, 1633 A and 1666 C. 4. Sequence according to claim 1 and 2, characterized by the mutations 1541 C, 1633 G and 1666 G. 5. Sequence according to claim 1 and 2, characterized by the mutations 1541 T, 1633 G and 1666 C. 6. Sequence according to claim 1 and 2, characterized by the mutations 1541 T, 1568 T 1633 A and 1666 C. 7. Sequence according to claim 1 and 2, characterized by the mutations 1541 C, 1568 C 1633 G and 1666 G. 8. Sequence according to claim 1 and 2, characterized by the mutations 1541 T, 1568 T 1633 G and 1666 C. 9. A method for determining disease predispositions, characterized in that the DNA of a subject is isolated and genotyped at least at one of the exchanged positions and subsequently compared with the reference DNA sequence, with possible combinations of variants from the individual mutation up to all possible combinations of all variants, including any absolute number of variants, are included. 12 10. The method according to claim 9, characterized in that the DNA of a subject is isolated and genotyped at least at position 1633. 11. The method according to claim 9, characterized in that the DNA of a subject is isolated and genotyped at least at the 3 positions 1541, 1633 and 1666. 12. The method according to claim 9, characterized in that the DNA of a subject is isolated and genotyped at least at the 4 positions 1541, 1568, 1633 and 1666. 13. The method according to claim 9, characterized in that the DNA of a subject is isolated and genotyped at least at the 7 positions 245, 565, 934, 1541, 1568, 1633 and 1666. 14. The method according to claim 9 or 12, characterized in that the positions 1541, 1568, 1633 and 1666 are genotyped. 15. The method according to claim 14, characterized in that at least 3 of the 4 positions 1541, 1568, 1633 and 1666 are genotyped. 16. A method for determining disease dispositions 9, 11 or 15, characterized in that positions 1541, 1633 and 1666 are genotyped. 17. The method according to any one of claims 9 to 16, characterized in that the genotyping is carried out by sequencing or by other methods which are suitable for the detection of variants. 18. The method according to any one of claims 9 to 17 for determining a disposition for high blood pressure and for blood pressure deviations from the norm and other cardiovascular diseases, including myocardial infarction and stroke; for determining a disposition for neuropsychiatric diseases such as depression, anxiety syndromes, attention deficit disorder with hyperactivity, eating disorders, eg anorexia nervosa and bulimia, or for disorders triggered by post-traumatic stress; for determining a disposition for diseases of the autonomic nervous system, such as Bradbury-Eggleston, Sky-Drager and Riley-Day syndrome, and selective noradrenergic and baroreceptor dispositions, or migraines; to determine a disposition for allergic diseases, especially asthma and atopic disorders; to determine a disposition for metabolic diseases such as obesity and family morbid obesity, 13 including a prediction of the weight range as such or a disposition for weight change, finally a prediction of the ratio of the body measurements as such, e.g. Express in the 'body mass index' (BMI). 19. The method according to any one of claims 9 to 17 for determining an individually different reactivity of the autonomic nervous system, in particular on endogenous and exogenous stress. 20. The method according to claim 19 for determining an individually different disposition for blood pressure and / or heart rate changes / deflections on endogenous and exogenous stress, or an individually different salt sensitivity / resistance. 21. The method according to any one of claims 9 to 17 for determining the course and severity of diseases, such as listed under claim 18, e.g. B. neuropsychiatric diseases such as depression and anxiety syndromes, cardiovascular diseases, including myocardial infarction and stroke, diseases of the autonomic nervous system and allergic diseases such as e.g. Asthma. 22. The method according to any one of claims 9 to 17 for determining a disposition for metabolic diseases such as obesity. 23. The method according to any one of claims 9 to 17 for predicting the survival after serious medical diseases, e.g. after myocardial infarction, heart failure and / or stroke. 24. Use of the sequence variants according to claim 1 to 8 for the development of therapeutic agents and / or lifestyle drugs. 25. Use according to claim 24 for the development of a new class of therapeutic agents which are directed to the beta2 receptor gene and which attack the 5 'regulatory region, the promoter region and the leader peptide, by regulating the transcription and translation and by influencing their efficiency , primarily by regulating expression. 26. Use according to claim 24 for the development of beta2-receptor agonists and - antagonists, in particular of individually specific beta2-receptor agonists and - antagonists. 14 27. Use according to one of claims 9 to 17 for predicting the individually different responsiveness to previously known, such as beta2-receptor ligands, as well as therapeutics developed in the future, also under claims 24 to 26, and the individually different responsiveness to the endogenous ligands adrenaline and noradrenaline. 28. Use according to one of claims 9 to 17 for predicting the individual habituation to medication (tachyphylaxis), as well as a different disposition for drug side effects. 29. Use according to one of claims 9 to 17 for optimizing the individual therapy or intervention directed at the beta2 receptor and its gene. 30. Use of the sequence variants according to claims 1 to 8 for the construction of genes or vectors, in particular for the development of pharmaceutically relevant substances. 31. Use according to claim 24 to 30 for the development of a diagnostic kit, or any method for genotyping. 32. Use according to claims 24 to 31 for the development of a diagnostic kit for predicting the individual responsiveness to various beta2 receptor agonists and antagonists, as well as to any newly developed beta2 active therapeutic agents, in particular also according to claim 25; to predict the therapeutic efficacy of pharmaceuticals whose mechanism of action includes changes in the beta2 receptor structure, regulation or expression; to predict individual responsiveness to the endogenous ligands epinephrine and noradrenaline; to predict individual habituation to drug administration - tachyphylaxis - as well as a different disposition for drug side effects; to optimize the individual therapy or intervention aimed at the beta2 receptor and its gene. 33. Use according to claims 1 to 8 and 9 to 32 for the development of in vitro (for example cell cultures) and in vivo (for example transgenic animals) test systems which express individual forms of the beta2-receptor gene, the test systems for examining the pathophysiology of Diseases of generally medically important characteristics with the participation of the beta2 prescription gene serve, as well as for the development and testing of individually specific therapeutic agents and 'lifestyle drugs' and of beta2-directed substances in general.