DE10004858A1 - Use of protein kinase inhibitor alpha - Google Patents

Abstract

The invention concerns the utilization of PKI alpha and/or the derivatives thereof to lower blood pressure, as beta antagonists and for the production of a medicament for the prevention and/or treatment of diseases in which a beta antagonist can be used. The invention also concerns the utilization of an inhibitor of PKI alpha or a derivative thereof to lower blood pressure, as beta agonist and for the production of a medicament for the treatment and/or prevention of diseases in which a beta agonist can be used.

Description

The present invention relates to the use of protein kinase inhibitor-alpha (PKI- alpha), the use of inhibitors of protein kinase inhibitor alpha and method for screening for beta agonists and beta antagonists.

In 1975 the phenomenon of "myocardial stunning" was first described. Because of that ter is the property of the myocardium after short-term ischemia due to coronary artery Occlusions to show long-lasting, reversible myocardial dysfunction, d. H. the Cardiac muscle does not show the expected contractility. This reversible state lasts for several hours and contradicted the previously widespread opinion among experts that a Ischemic state of more than 45 minutes irrevocable with my death ocytes goes hand in hand.

The formation of Sauer was used as a pathogenicity mechanism for the "myocardial stunning" substance radicals, calcium overload, dysfunction of the sarcoplasmic reticulum and Troponin proteolysis discussed (Bolli; Mechanism of myocardial "stunning". Circulation 1990: 723-38; and Wijns, Vatner, Camici. Hibernating myocardium. New England Journal of medicine. 1998. 173-81). However, each of these mechanisms is not suitable, the pheno fully described (Duncker et al .; Myocardial stunning remainig questions. Cardiovasc Res 1998; 38: 549-558).

The protein kinase A present in the heart (henceforth also referred to as PKA), which the myocar regulated contractility by means of phospholamban and phosphorylation of troponin I, is controlled by receptors that are coupled to the adenylate cyclase, which in turn is regulated by catecholamines. The enzyme activity is both by protein kinase Inhibitors (hereinafter referred to as PKI) as well as regulated by cAMP. In the heart there are two forms of protein kinase inhibitors: the alpha and the beta form (PKI- alpha and PKI-beta). Both act as competitive inhibitors in that the Cα and Cβ  Subunits of active protein kinase A are blocked. The α shape is six times more effective than the β form.

A protein kinase inhibitor known in the art is the protein kinase inhibitor alpha (also referred to herein as PKI-alpha). PKI-alpha is 76 amino acids in length on, the essential for the inhibitory effect of the primary sequence on the Range of amino acids 14 to 22 appears to be limited. This sequence is in the menu Pork, pork, chicken, rat and mouse are 100% identical. At the level of the mRNA the 5 'end of the total porcine mRNA shows a sequence homology of approximately 90% compared to that of the human, mouse, rat and chicken genome chen. A recombinant PKI-alpha comprises only amino acids 14 to 22 of the complete PKI-alpha amino acid sequence is cell permeable and available from Calbiochem (Pro product number # 476485).

The porcine cDNA coding for PKI-alpha has a second one at its 5 'end open reading frame encoding a 9 amino acid polypeptide, and several ver separated poly (A) signal sequences at its 3 'end (GenBank #ban kit256111AF132737)

The present invention has for its object new uses for PKI-alpha and to provide for an inhibitor of PKI-alpha.

According to the invention, the object is achieved by using PKI-alpha and / or its derivatives for lowering blood pressure.

In a further aspect of the invention, the object is achieved according to the invention by Use of PKI-alpha and / or its derivatives as a beta antagonist.

In a still further aspect of the invention, the object is achieved through the use of PKI-alpha and / or its derivatives for the manufacture of a medicament for prevention tion and / or treatment of diseases in which a beta-antagonist is used can.  

In the use according to the invention it can be provided that the PKI-alpha has a SEQ ID No. 2 amino acid sequence shown comprises.

In one embodiment of the uses according to the invention, the PKI-alpha is encoded from a nucleic acid that corresponds to the coding sequence of the sequence shown in SEQ ID No. 1 shown Se sequence or one resulting from the degeneracy of the genetic code or corresponds to a nucleic acid hybridizing to it.

In the uses according to the invention it can also be provided that the PKI-alpha in a shortened form is used.

In a preferred embodiment of the uses according to the invention, pre-can can be seen that the PKI-alpha amino acids 14 to 22 of the PKI-alpha sequence, in particular the sequence of PKI-alpha according to SEQ ID No. 1 or SEQ ID No. 2 includes.

In particularly preferred embodiments of the uses according to the invention provided that the PKI-alpha or its derivative for the treatment and / or prevention of Diseases are used, which are selected from the group, the cardiac arrhythmia genes, hyperkinetic heart syndrome, angina pectoris, myocardial infarction, acute myocardium farct, heart failure, arterial hypertension, essential and renal hypertension, portal vein high pressure, bleeding from esophageal varices, pheochromocytoma, overdose of beta Sympathomimetics and choline receptor blockers, Glaucoma simplex, hyperthyroidism, Thy reotoxicosis, migraines, essential tremor, arousal tremor and alcohol withdrawal syndrome includes.

In further particularly preferred embodiments of the use according to the invention gene is provided that the PKI-alpha or its derivative for post-infarct medication, Pro prophylaxis of the Reinfarktes and / or used as a sedative.

According to the invention, the object is also achieved by using an Inhi bitors of PKI-alpha or a derivative thereof for increasing blood pressure.  

In a further aspect, the object is achieved by using an inhibitor of PKI-alpha or a derivative thereof as a beta agonist.

In a still further aspect, the object is achieved by using an inhibi PKI-alpha or a derivative thereof for the manufacture of a medicament for the Treatment and / or prevention of diseases in which a beta agonist is used can be.

In a preferred embodiment of the uses according to the invention, there is seen that the inhibitor affects the transcription of the PKI-alpha.

In the use according to the invention it can further be provided that the PKI-alpha is a PKI-alpha is as described herein.

In a further embodiment of the uses according to the invention, that the inhibitor of PKI-alpha comprises an amino acid sequence selected from the Group containing sequences with SEQ ID No. 3 and SEQ ID No. 4 includes.

In particularly preferred embodiments of the uses according to the invention provided that the inhibitor of PKI-alpha for the treatment and / or prevention of ill units selected from the group consisting of heart failure, heart attack, includes chronic and acute heart failure and hypotension.

In a further aspect, the object is achieved by a method according to the invention for screening for agents for lowering blood pressure, beta-antagonists and / or agents for the treatment and / or prevention of diseases in which a beta-antagonist can be used, that

• A mixture of a beta receptor, a beta agonist of beta Receptor and a candidate beta antagonist is provided, and
• - It is determined to what extent the candidate beta-antagonist Binding behavior of the beta agonist to the beta receptor changes.

In one embodiment of the method according to the invention it is provided that the under Change in binding behavior observed under the influence of the candidate beta-antagonist tens of the beta agonist to the beta receptor is compared with that under the influence of beta antagonists, as described herein, observed changes in binding holding the beta agonist to the beta receptor.

In a preferred embodiment of the method according to the invention, that the beta receptor is selected from the group consisting of the beta1 and beta2 receptors sums up.

In yet another embodiment of the method according to the invention, that the beta agonist is a sympathomimetic, especially one that is selected is from the group that includes noradrenaline, adrenaline, dopamine and dobutamine.

Finally, it can be provided in the method according to the invention that the beta Agonist is a beta agonist as described herein.

In a still further aspect, the object is achieved by a method for screening for agents for lowering blood pressure, beta-agonists and / or agents for the treatment and / or prevention of diseases in which a beta-agonist can be used, wherein

• A transcription system for a beta-antagonist is provided,
• A candidate beta agonist is added to the transcription system and
• - It is determined to what extent the transcription for by the beta agonist changes the beta antagonist.

In a preferred embodiment of the method according to the invention, that the change in transcription observed under the influence of the candidate beta agonist  of the beta antagonist is compared with that under the influence of the beta Agonists, as described herein, observed changes in beta transcription Antagonists.

In a further embodiment of the method it is provided that the beta-antagonist such is as described herein.

In yet another embodiment of the method according to the invention, that the beta-antagonist is obtainable by one of the methods according to the invention.

The present invention is based on the surprising finding that PKI-alpha is a negative inotropes, i.e. H. has an adverse effect on myocardial function. This Effect comes about by an inhibition of protein kinase A, which for phosphoryly phospholamban and troponin is responsible (so-called beta-adrenergic signal trans production). As a result of the negative inotropic effect of PKI-alpha, the reverse is done Inhibition of PKI-alpha improves myocardial function, especially in heart failure sert.

The uses according to the invention are all based on the Er described above Knows that PKI-alpha has a concentration-dependent, negative inotropic effect, and is involved in beta-adrenergic signal transduction. This results in one Aspect that PKI-alpha for lowering blood pressure or for the manufacture of a drug here can be used. In another aspect it follows that PKI-alpha as beta antagonist can be used. The latter is due to the fact that the effect of Catecholamine binding beta receptors on the contractility regulating protein ki nose A can be counteracted by the protein kinase inhibitors, this one Show beta-antagonistic effects.

Due to the beta-antagonistic effect of the PKI-alpha, it is also suitable for use here Provision of medicines for the treatment of diseases in which a general beta antagonist is used. In other words: due to the beta-adrenergic effect  PKI-alpha can use this or derivatives thereof instead of or in addition to the beta antagonists used in medication. Basically there are no restrictions, so that with PKI-alpha or with egg In addition to the medication containing this, a replacement for any Antagonists is provided as a pharmaceutical drug.

Because the PKI-alpha not only in muscle tissue, but also in nerve tissue, in particular in the brain that is expressed are also neurophysiological indication fields for the PKI-alpha and inhibitors thereof in the context of the present invention. Example We would like to refer to migraines for which beta-antagonists are used to treat which can be replaced by PKI-alpha according to the invention. It results for the subject man that any tissue that expresses PKI-alpha and is pathological or changed Develops conditions, therapeutic according to the invention with PKI-alpha and inhibitors of the same or can be treated preventively.

There are a number of advantages with the uses according to the invention. So will with the use of the PKI-alpha and an inhibitor thereof the group of pharma Zeutisch effective compounds significantly expanded, in particular to the extent that a completely new class of compounds in these therapeutically very important indicators ons territory is introduced. In addition, the use of PKI-alpha and inhibitors of which also advantageous in that they are very early in the cascade of beta-adrenergic Apply signal transduction and therefore not the unwanted and with various disadvantages connected down regulation of the beta receptors. These advantages will be are further explained below using heart failure.

Heart failure belongs in the western industrialized countries due to its high prevalence (1-1.5%) and morbidity of the adult population and its correspondingly high costs on the most important diseases (Sharpe N. et al .; Lancet 352 (suppl I), 3-7 (1998)). To A number of medications are used in her therapy, but only a few of them ge really, d. H. effective for many years to come. These primarily include β- blocking substances and ACE inhibitors (Cleland, J; Lancet 352 (suppl I), 1-2 (1998)). In contrast to this, the calcium antagonists that are still used (e.g. Adalat-  Nifedipine) have been discredited (Prescrire Int 7 (35), 90-91 (1998); and Ishibashi, Y. et al .; Clin Exp Pharmacol Physiol 26 (5-6), 404-10 (1999)). Especially their acute effects, which the Mortality in treated patients appear to increase caution when using of these drugs. In contrast, β-blockers appear to have significant advantages particular regarding the mortality to have for the affected patient. Unfortunately act these drugs via the β1 and β2 receptors, the so-called down regulation are subject to and therefore miss their place of action (Massie, B .; Lancet 352 (suppl I), 29-33 (1998). There is therefore an increasing search for substance classes that know their place of action ter above the β-adrenergic signal transduction cascade. By inhibiting the Protein kinase A, especially by intracoronary application, appears to be the PKI-alpha, in particular the amino acids 14-22 from belonging to the desired class of substances.

In this context, an inhibitor of PKI-alpha appears particularly interesting, especially the PKI-alpha inhibitor encoded by the second open reading frame of the PKI- alpha (PKI-alpha-I). As required, both the beta and antagonistic as well as beta agonistic effects through the use of PKI alpha or an inhibitor thereof.

The amino acid sequence of PKI-alpha is shown in SEQ ID No. 1 together with which you code the nucleic acid sequence and as an amino acid sequence as such in SEQ ID No. 2. It is apparent to the person skilled in the art that the PKI-alpha is not based on that PKI-alpha is limited, which as the sequences SEQ ID No. 1 and SEQ ID No. 2 sequences shown having. Rather, under a PKI-alpha, any of these will be the activity of the protein Understanding kinase A influencing protein kinase inhibitor. In this respect, the concept of Protein kinase inhibitor based on a functional definition herein.

As a result of the functional definition of the term protein kinase Inhibitors is also the one in SEQ ID No. 1 nucleic acid sequence is only an example for a sequence encoding a protein kinase inhibitor. Other sequences arise from the degeneracy of the genetic code.  

In its native form, the protein kinase inhibitor-alpha has a length of 76 amino acids open up. In fact, the range relevant to the inhibitory effect extends Primary sequence of amino acids 14 to 22, so that even truncated ("truncated") forms of Protein kinase inhibitors, in particular of PKI-alpha, can be used according to the invention can. The use of PKI-alpha, whose Se sequence is limited to amino acids 14 to 22 of the complete amino acid sequence. This shortened PKI-alpha is membrane-permeable or membrane-permeable, i.e. H. he can over a cytoplasmic membrane enters the intracellular space and there the intracel lular compartments, in particular the sarcoplasmic reticulum and the protein kinase A, reach. Thus PKI-alpha and in particular a shortened form can be very special ders the amino acids 14 to 22 of the complete primary sequence of the PKI-alpha, as placed in SEQ ID No. 1 or SEQ ID No. 2, comprehensive shortened form intracoronary, intra be given peritoneally or intravenously.

It is within the skill of the artisan that PKI-alpha as well Inhibitor may be in a modified form. Such a modification can, for example, in In connection with the requirements of galenics. Other modifications can the pharmacological and toxicological properties.

Formulations comprising PKI-alpha or an inhibitor thereof can be the usual pharmaceutical excipients and ingredients such as pharmaceutically acceptable carriers and buffer.

Beta-blockers, ie beta-antagonists, and thus PKI-alpha and its derivatives according to the invention are used, for example, as or for

• 1. Antiarrhythmic drugs (irregular heartbeat)
• 2. Hyperkinetic heart syndrome
• 3. Anti-anginal drugs (angina pectoris)  
• 4. Improving the prognosis in patients with myocardial infarction (post-infarction Medication), acute myocardial infarction and prophylaxis of the pure color
• 5. Therapy for heart failure
• 6. Antihypertensives (arterial hypertension, essential and renal hypertension)
• 7. Portal vein hypertension, bleeding from esophageal varices
• 8. pheochromocytoma (only applicable after previous alpha blockade)
• 9. Overdose of β-sympathomimetics and m cholinoceptor blockers
• 10. Glaucoma on the eye (Glaucoma simplex)
• 11. Hyperthyroidism and thyrotoxicosis
• 12. Therapy of central nervous diseases such as migraines and as a sedative (central damping effect) (essential tremor, excitation tremor, alcohol Withdrawal syndrome)

The use according to the invention of an inhibitor of PKI-alpha (hereinafter also as follows referred to as PKI-alpha-I) is again based on the surprising finding that PKI- alpha has a concentration-dependent, inotropic effect and on the beta-adrenergic Signal transduction is involved. In the light of these properties of PKI-alpha, an Inhi bitor of PKI-alpha leads to a modification and reversal of the effects of PKI-alpha As a result, an inhibitor of PKI-alpha for increasing blood pressure or one here can be used for suitable medication. After the inhibitor of PKI-alpha on acts as an inhibitor of beta-adrenergic signal transmission, it acts as a beta Agonist and can be used accordingly. This use generally includes Use in the manufacture of a medicament for the treatment and prevention of Diseases where beta agonists can be used: Thus, an inhibitor of PKI-alpha in addition to the previously used beta agonist in medication  be used. Basically there are no restrictions, so that with the inhibitor of PKI-alpha now a replacement for any beta agonist as Drug containing pharmaceutical active ingredient is provided.

The term inhibitor of PKI-alpha is again a functional definition underlying. An inhibitor of PKI-alpha is thus any compound that has the effect inhibited by PKI-alpha, whereby the extent to which the inhibition of PKI- Alpha is assigned to which chemical compound class the PKI-alpha inhibitor is assigned and on the basis of what mechanism the inhibitor of PKI-alpha is Effect.

A mechanism that can be the basis of the action of an inhibitor of PKI-alpha is the inhibition of the transcription of PKI-alpha. The gene product of the second open reading frame of the PKI-alpha gene, for example, acts as an inhibitor in this sense. It appears from Wang X. et al .; that the gene product can be the active agent. Mol Pharm 54, 514-524 (1998). The amino acid sequence of this human gene product was

Met Trp Ile Phe Gly Ser Asn Asp

and is as SEQ ID No. 3 together with the nucleic acid sequence encoding them herein taken.

The corresponding pork sequence is

Met Trp Ile Phe Val Ser Asn Asp

and is as SEQ ID No. 4 together with the nucleic acid sequence encoding them herein taken.  

The difference in the form of a difference between the two types of human and pig Amino acid exchange at position 5 of the amino acid sequence is based on an out exchange at the level of the nucleic acid, whereby position 2 of the fifth codon is changed. in the In the human sequence, glycine is found as the fifth amino acid, whereas the se in the case of the pork sequence is valine. The corresponding base from swap is a change from G (human) to T (pig).

It is obvious to the person skilled in the art that deviations from those in SEQ ID No. 3 and SEQ ID No. 4 disclosed sequences are possible as long as that derived from the sequences Derivative is still an inhibitory one for PKI-alpha, in particular its transcription Has effect.

Beta agonists and thus inhibitors of PKI-alpha, as disclosed herein, can be used for

• 1. Increase in contractility of the myocardium (desirable for severe heart failure ciency or in heart attack patients (chronic heart failure and acute heart attack efficiency)
• 2. Hypotension

It is obvious to the person skilled in the art that the uses according to the invention in the framework men of therapeutic and preventive treatments can be used, esp especially to people who need such treatment.

The methods according to the invention are methods by means of which verb be determined or made available that as PKI-alpha, d. H. as inhibitors of the Pro Tinkinase A or as inhibitors thereof can be used, especially in Zu associated with one or more of the diseases or indications disclosed herein fields. The methods can also be directed to the fact that starting from a large number  of connections one or more of them with a corresponding property to be averaged ("screening"). A connection that may match one de property, is also referred to herein as a "candidate" compound.

In the method according to the invention for determining or screening a broad Most sensible beta-antagonistic compound, the receptor provided as isolated molecule, for example in solution or immobilized. However, it is also in the Within the scope of the present invention, the receptor is present in a biological system lies. Such a biological can be an in vitro system or an in vivo system. On A typical in vitro system is a translation or transcription system. A typical one in vivo system is a cellular system, for example a cell, in particular a myocardial or nerve cell that contains the genetic information for PKI-alpha. A cellular system but can also be a cell into which, preferably by means of genetic engineering metho the, a nucleic acid coding for PKI-alpha or a part thereof or its or de ren expression-controlling nucleic acid is introduced. When using the in vivo and in in vitro systems can be based on the stress inducibility of the PKI alpha can be used.

In the method according to the invention it can further be provided that the effectiveness a candidate antagonist is evaluated based on a comparison of the we PKI-alpha and its derivatives as described herein. This can new at a very early stage of the screening or development process tiger beta-antagonists and beta-agonists quantify the effectiveness of the gete constant connections. The method according to the invention can also provide that a beta agonist as described herein is used.

In the method according to the invention for determining or screening a broad Most beta-agonist compound can be provided by the beta agonist provided isolated molecule, for example in solution or immobilized. However, it is also in the Framework of the present invention that the candidate agonist in a biological system  is present. Such a biological system can be an in vitro system or an in vivo System. A typical in vitro system is typically a cell-free translation or transcription system. A typical in vivo system is a cellular system For example, a cell, especially a myocardial or nerve cell, which genetic information on for the candidate beta agonist. A cellular system can also be a Be cell in which, preferably by means of genetic engineering methods, for the candy data beta agonists or a part thereof encoding nucleic acid or its Expression-controlling nucleic acid is introduced.

The transcription system for egg used in the process according to the invention A beta-antagonist can be an in vivo system or an in vitro system. A typi The in vitro system is a translation or transcription system. A typical in Vivo system is a cellular system, for example a cell, especially a myocardial or nerve cell that contains the genetic information for PKI-alpha. A cellular system but can also be a cell into which, preferably by means of genetic engineering metho the nucleic acid coding for PKI-alpha or a part thereof or its or its Expression-controlling nucleic acid is introduced. When using the in vivo and in in vitro systems can be based on the stress inducibility of the PKI alpha can be used.

It is also within the scope of the method according to the invention that the transcription system for a beta-antagonist also the candidate beta-agonist or the coding for it or comprises nucleic acid controlling its expression. Furthermore, it is within the The method according to the invention that beta-antagonists and beta-agonists, as described herein are disclosed and described or determined according to one of the methods disclosed herein or can be determined as such, used therein.

In the method according to the invention it can further be provided that the effectiveness a candidate beta agonist is evaluated based on a comparison of the un Under the influence of the candidate beta agonist observed change in the transcription of the  beta-antagonists with that under the influence of a beta-agonist as described herein is observed change in transcription of the beta-antagonist. This can already lead to a very early stage of screening or the development process of novel beta Agonists can quantify the effectiveness of the tested compounds.

The invention is further illustrated by the following figures, examples and the sequence listing are illustrated, from which further features and advantages of the invention result can. It shows:

Fig. 1 shows the function of protein kinase A and PKI-alpha in beta-adrenergic signal transduction: More precisely, the signal transduction cascade of the β1 and β2 receptors (β1 and β2 AR) is shown, which is via a, β and yG protein - Subunits as well as the adenylate cyclase (AC) and cAMP increases activate the protein kinase A. The protein kinase A can be inhibited by the PKI-alpha, whereby the effects of the protein kinase A can be antagonized. Protein kinase A phosphorylates in particular phospholamban, which explains the positive inotropic effect.

Fig. 2 shows the decrease in the function of the left ventricle after administration of PKI-alpha expressed as dp / dt and external cardiac work (EHW).

EXAMPLES example 1 Studies on the effect of PKI-alpha on a working rat heart

The hearts of a total of six Sprague Dawley rats weighing approximately 300 g were isolated and, as in Schulze et al. (Schulze et al .; Circulation 1990: 959-69) described perfused. Hemodynamic parameters including contractility, external cardiac work (EHW), coronary and aortic flow were recorded approximately 15 minutes after antegrade perfusion. Cell permeable, recombinant PKI-alpha (amino acids 14 to 22, Calbio chem # 476485) was then administered. The compound was infused directly into the atrium cannula using a motor pump. The infusion rate was continuously adjusted to the ejection volume (the sum of the aortic and coronary flow) and increased at five minute intervals in order to achieve the final PKI-alpha concentrations in the perfusate of 0.5, 1.0 and 2.0 µmol / l . The results relating to the external heart volume and the course of dp / dt _max under these test conditions are shown in FIG. 2.

The infusion of the shortened PKI-alpha caused a pronounced, reversible, concentrate ons dependent (up to 2 µmol) depression of the myocardial function, which was comparable to the dysfunction observed after short coronary occlusion. The measured decrease in external heart work and dp / dt-max was in the paired t-test (narrowly: "paired t-test") for everyone tested concentrations significantly. Perfusion with the shortened PKI-alpha resulted in a 43% reduction in external cardiac work and a decrease in left ventricle function, expressed as dp / dt-max, by 53%.

It follows from these studies that the shortened form of the PKI-alpha is suitable for lowering blood pressure and generally for use as a new form or class of substance a beta antagonist.

Example 2 Investigations on cell cultures

On cell cultures of myocytes (e.g. C9H2 cells, available from American Type Culture Collection), the protein kinase shortened to amino acids 14 to 22 Given inhibitor alpha and the activity of protein kinase A determined. It became a do sis-dependent decrease in activity.

The 9 amino acid long PKI-alpha inhibitor (the Gene product of the second open reading frame of the PKI gene in humans and in humans Pig) and the expression of PKI-alpha and the activity of protein kinase A certainly. There was an increase in the dose dependent on the PKI-alpha inhibitor Activity of the PCA determined.  

Cell cultures of myocytes were analyzed with an expression plasmid with the sequence of PKI- alpha, as described, for example, by Olsen (Olsen et al .; Endocrinology 5; 1246-1256 (1991)) transfected and the expression of the PKI-alpha determined. At the same time, the activity of Protein kinase A, which is specifically inhibited by PKI-alpha, was determined. (Protein kinase A is Contained in both striated and smooth muscle cells (Wang X. et al .; Mol Pharm 54, 514-524 (1998))). Different concentrations of the Pro teinkinase inhibitor-alpha-inhibitor (PKI-alpha-I) given. Expression of the PKI-alpha was decreased depending on the concentration of the PKI-alpha inhibitor.

The disclosure of the various references cited herein is hereby disclosed by Be full admission.

Those disclosed in the foregoing description, claims and drawing Features of the invention can be used both individually and in any combination for the Realization of the invention in its various embodiments may be essential.  

SEQUENCE LOG

Claims (25)

1. Use of PKI-alpha and / or its derivatives for lowering blood pressure. 2. Use of PKI-alpha and / or its derivatives as a beta antagonist. 3. Use of PKI-alpha and / or its derivatives for the manufacture of a medicament mentes for the prevention and / or treatment of diseases in which a Antagonist can be used. 4. Use according to one of claims 1 to 3, characterized in that PKI- alpha one in SEQ ID No. 2 amino acid sequence shown comprises. 5. Use according to one of claims 1 to 3, characterized in that the PKI-alpha is encoded by a nucleic acid that corresponds to the coding sequence of the SEQ ID No. 1 shown sequence or one resulting from the degeneracy of the genetic code or a hybridizing nucleic acid corresponds. 6. Use according to one of claims 1 to 5, characterized in that the PKI-alpha is used in a shortened form. 7. Use according to claim 6, characterized in that the PKI-alpha the Ami no acids 14 to 22 of the PKI-alpha sequence, in particular the sequence of PKI-alpha according to SEQ ID No. 1 or SEQ ID No. 2 includes. 8. Use according to one of claims 1 to 7, characterized in that the PKI-alpha is used to treat and / or prevent disease are selected from the group, cardiac arrhythmia, hyperkinetic heart syndrome, angina pectoris, myocardial infarction, acute myocardial infarction, heart failure, arterial hypertension, essential and renal hypertension, portal hypertension, bleeding  from esophageal varices, pheochromocytoma, overdose of beta Sympathomimetics and choline receptor blockers, Glaucoma simplex, Hyperthyreo se, thyrotoxicosis, migraines, essential tremor, excitatory tremor and alcohol Withdrawal syndrome includes. 9. Use according to one of claims 1 to 8, characterized in that the PKI-alpha for post-infarct medication, prophylaxis of the reinfarcts and / or as Beru detergent is used. 10. Use of an inhibitor of PKI-alpha or a derivative thereof for blood pressure increase. 11. Use of an inhibitor of PKI-alpha or a derivative thereof as beta Agonist. 12. Use of an inhibitor of PKI-alpha or a derivative thereof for the manufacture treatment of a medicament for the treatment and / or prevention of diseases, where a beta agonist can be used. 13. Use according to one of claims 10 to 12, characterized in that the Inhibitor affects the transcription of PKI-alpha. 14. Use according to one of claims 10 to 13, characterized in that the PKI-alpha is a PKI-alpha according to one of claims 1 to 9. 15. Use according to one of claims 10 to 14, characterized in that the Inhibitor of the PKI-alpha comprises an amino acid sequence which is selected from the Group containing sequences with SEQ ID No. 3 and SEQ ID No. 4 includes. 16. Use according to one of claims 10 to 15, characterized in that the PKI-alpha inhibitor for the treatment and / or prevention of diseases ver who are selected from the group consisting of heart failure, heart attack, includes chronic and acute heart failure and hypotension.   17. A method of screening for agents for lowering blood pressure, beta-antagonists and / or agents for the treatment and / or prevention of diseases in which a beta-antagonist can be used, characterized in that

• a mixture of a beta receptor, a beta agonist of the beta receptor and a candidate beta antagonist is provided, and
• - It is determined to what extent the binding behavior of the beta agonist to the beta receptor changes due to the candidate beta antagonist.

18. The method according to claim 17, characterized in that the under the influence of Candidate beta-antagonists observed changes in the binding behavior of the be ta agonsites to the beta receptor is compared to that under the influence of the be ta antagonists according to one of claims 1 to 9 observed change in the bin behavior of the beta agonist at the beta receptor. 19. The method according to claim 17 or 18, characterized in that the beta receptor is selected from the group comprising beta1 and beta2 receptors. 20. The method according to any one of claims 17 to 19, characterized in that the be ta agonist is a sympathomimetic, especially one that is selected from the group including noradrenaline, adrenaline, dopamine and dobutamine. 21. The method according to any one of claims 17 to 20, characterized in that the be ta agonist is a beta agonist according to any one of claims 10 to 16. 22. A method of screening for agents for lowering blood pressure, beta-agonists and / or agents for the treatment and / or prevention of diseases in which a beta-agonist can be used, characterized in that

• a transcription system for a beta-antagonist is provided,
• a candidate beta agonist is added to the transcription system and
• - It is determined to what extent the beta-agonist changes the transcription for the beta-antagonist.

23. The method according to claim 22, characterized in that the under the influence of Candidate beta agonists observed change in beta transcription Antagonists are compared with those under the influence of the beta agonist according to ei Nem of claims 11 to 16 observed change in the transcription of the beta Antagonists. 24. The method according to claim 22 or 23, characterized in that the beta Antagonist is one according to one of claims 1 to 9. 25. The method according to any one of claims 22 to 24, characterized in that the beta Antagonist is obtainable by a process according to one of claims 17 to 21.