WO2003057912A2 - Molecular trichogram - Google Patents

Abstract

The invention relates to a method for diagnosing impaired hair growth or for determining the probability of impaired hair growth (predisposition) in mammalians. According to the invention, the expression of at least two cyclically expressed genes in a sample is analyzed and (a) one of the two genes has the maximum expression peak during the anagen phase while (b) the second gene has the maximum expression peak during the catagen phase or during the telogen phase. The invention further relates to a method for diagnosing the effectiveness of the treatment of impaired hair growth in mammalians, according to which the expression of at least two cyclically expressed genes in a sample is analyzed and (a) one of the two genes has the maximum expression peak during the anagen phase while (b) the second gene has the maximum expression peak during the catagen phase or during the telogen phase. Preferably, the expression of further genes with a cyclic expression curve is analyzed according to the inventive method. The invention also relates to a method for isolating inhibitors of impaired hair growth and for improving such inhibitors. Finally, the invention relates to methods for producing drugs or medicinal products in which the identified or improved inhibitor is formulated together with a pharmaceutically acceptable excipient or diluent.

Description

 Molecular trichogram

The invention relates to a method for diagnosing a hair growth disorder or for determining the likelihood of a hair growth disorder (predisposition) in a mammal, wherein the expression of at least two cyclically expressed genes in a sample is analyzed and wherein (a) one of the genes is the maximum expression level during the Anagen, while (b) the second gene has the maximum expression level during catagen or during telogen. In addition, the invention relates to a method for diagnosing the effectiveness of treating a hair growth disorder in a mammal, analyzing the expression of at least two cyclically expressed genes in a sample and wherein (a) one of the genes has the maximum expression level during anagen, during the (b) second gene has the maximum level of expression during catagen or telogen. The expression of further genes with a cyclic expression course is preferably analyzed in the method according to the invention. In addition, the invention relates to methods for isolating inhibitors of hair growth disorders and for improving such inhibitors. Finally, the invention relates to processes for the production of pharmaceuticals or medical products, in which the identified or improved inhibitor is formulated with a pharmaceutically acceptable carrier or diluent.

A number of prior art documents are cited in the description. The disclosure content of these documents, including the manufacturer's instructions for use, is hereby incorporated by reference.

The skin and its appendages are subject to constant renewal. A special feature of the hair follicle is that the renewal here is not continuous, but cyclical. The so-called hair cycle. runs in three phases. The The growth phase is called anagen and lasts about 2-6 years for human hair, during which the hair grows continuously. The hair growth rate is about 0.3 mm a day. After this phase, the hair goes into a transition phase (catagen), which lasts about 1-2 weeks. This is followed by the resting phase (telogen), which lasts several months and at the end of which the hair is expelled. In humans there are about 100,000 hairs on the scalp, of which about 85% are in the anagen, 1% in the catagen and about 14% in the telogen; this means that normal hair loss can be estimated at around 100 hairs per day. The phases of the hair cycle are shorter with the mouse.

At birth, the human body is covered with vellus hair (wool hair), on the head these hair transform into so-called terminal hair from childhood. In the young adult, the coat is formed in a "mature" form, i.e. there are hardly any wool hair left; With terminal hair, the body hair can be distinguished from the bristle hair (eyebrows, eyelashes, nose, ears) and the long hair on the head, beard, axils and genital area. Over the years, the long hair of the scalp, especially in men, is subject to a gradual change back to wool hair, which is known as retrograde metamorphosis. This reverse transformation leads to the appearance of the bald head, significantly promoted by male sex hormones (androgens). This is caused by the gradual prolongation of the resting phase and the shortening of the growth phases of the scalp hair. The wool hair that remains on the face, especially in women, is transformed into terminal hair with increased androgen levels, thereby causing the image of hirsutism. This can be due to hormonal disorders, predisposition or age. The prevalence of hair growth disorders requires the use of meaningful diagnostic procedures for the condition of the hair cycle.

The so-called trichogram is currently used to diagnose hair growth disorders in humans. This is based on the morphological analysis of the hair shafts. Epilating a tuft of hair from the scalp can one can conclude from the microscopic structure of the hair root on the cycle phase of the hair. Although this examination method is possible without any particular technical effort, the condition of the hair cycle or the modification of the hair cycle by certain substances could only be determined by means of morphological or enzymatic parameters that are difficult to quantify. Recently, however, the procedure has been performed less and less because of the relatively low diagnostic value.

Therefore, there is still a need for meaningful diagnostic methods which on the one hand predict an impending hair growth disorder, in particular baldness, but on the other hand also allow an analysis of the effectiveness of the course of treatment of a hair growth disorder. The object of the present invention was to provide such methods.

The invention thus relates to a method for diagnosing a hair growth disorder or for determining the likelihood of a hair growth disorder (predisposition) in a mammal, the expression of at least two cyclically expressed genes in a sample being analyzed and (a) one of the genes during the maximum expression level of the agent, while (b) the second gene has the maximum expression level during catagen or during telogen.

The term "predisposition" for hair growth disorder in a mammal preferably encompasses genetic as well as environmental, nutritional and iatrogenic conditions that can lead to hair growth disorder in a mammal.

The term "cyclically expressed gene" relates to a gene which has different levels of expression during the different stages of the hair cycle. These levels of expression are quantitatively comparable and ideally identical for different hairs from the same sample which are in the same stage of the hair cycle. As is known the hair cycle is divided into three stages, which are referred to as anagen, catagen and telogen, for example, if hair is epilated from one place on a subject's head, cyclic expressed the same genes from hair in telogen synthesized a comparable amount of mRNA. In the method according to the invention, hair is preferably removed from the same body regions. In another preferred embodiment, the amount of mRNA synthesized serves as a measure of the level of expression. In a further preferred embodiment, the amount of translated or post-translationally modified protein is measured. The term "anagen" as used in the present invention encompasses the early, middle and late anagen as well as the transition phase from anagen to catagen. In particular, the phases named as anagen I-VI and catagen I are included (cf. Müller -Rover et al., J. Invest. Dermatol. 117 (2001), 3-15). The term "catagen" as used in the present invention includes the phases referred to as catagen II-VIII (see FIG. Müller-Rover, loc. Cit.). The term "telogen" as used in the present invention encompasses the phase referred to as telogen (see Müller-Rover, loc. Cit.). The invention provides a new method which is easy to use with molecular biological methods and which provides the method mentioned above The analysis of the expression level of only two genes makes it possible to diagnose the predisposition for a hair growth disorder, whereby one of the genes must have its maximum expression level during anagen, while the other gene has its maximum expression level during catagen or during telogen In which sub-phases of the stated stages the maximum expression level occurs is of secondary importance, for example the gene which has its maximum expression level in the anagen can be in the early anagen, in the middle anagen, in the late anagen or also in the transition phase from Have anagen to catagen te fluctuations possible with regard to the exact point in time of the maximum expression level of the same gene. For example, a gene that has its maximum expression level in the borderline between early and middle anagen in a sample of a mammal, in particular a mouse or a human, in another sample of the same mammal, which was taken at the same time or at a different time, the have maximum expression level in the middle anagen. This variation can also occur in samples from a mammal another species or from another mammal of the same species. In the mammals mentioned, the fluctuations can occur with the same hair growth disorder or even in the absence of a predisposition for a hair growth disorder. The border area between early and middle anagen is only to be understood as an example from the multitude of possibilities in which certain time-varying ranges can occur.

The temporal demarcation between anagen, catagen and telogen is established in the state of the art and researched in particular on humans and mice; see e.g. Pause, op. Anagen in the mouse takes about 16 days, catagen about 3 days and telogen about 6 days (according to the results of other groups about 9 days). This demarcation made in the prior art on the basis of morphological criteria is also used in the present invention.

The - preferably relative - level of expression of the genes from the sample is compared with a standard which, for example, represents the level of expression of the same gene in a corresponding mammal and wherein the mammal has no hair growth disorder or, less preferably, another hair growth disorder. Alternatively, the expression level can be compared with another standard, for example the expression level of a so-called house-keeping gene. Examples of house-keeping genes are mitochondrial genes, ubiquitin genes, polymerase genes or nuclear llamine genes. It is further preferred that the standard and the sample have been taken from corresponding samples of the body. In addition, it is preferred that the donors of the sample and the standard are of the same gender and optionally also have the same age (+/- 5 years). The standard of course no longer has to be a real sample, but is preferably a molecular trichogram stored on a data carrier, which can have a different complexity and be adapted to the needs. In this context, it is also recommended that the sample and the standard have been prepared with comparable methods or with methods providing comparable results in order to ensure the comparability of the results. Alternatively, the expression levels of the at least two cyclically expressed genes are compared directly with one another, it being possible to make a diagnosis as mentioned above from the relative value determined in this way. In this case, the corresponding expression levels of the genes for normal hair growth must be known. Alternatively, a known level of expression of these genes can serve as a standard in the event of hair growth disorder.

According to the invention, the hair cycle can thus be described quantitatively in molecular detail in a hitherto unknown detail by determining the activity of individual genes. The principle of this procedure is to select genes that are only active in one of the stages of the hair cycle. This active stage (e.g. anagen) correlates with the maximum expression level, the analysis of which is the basis for the method according to the invention. If you now measure the level of expression of such genes in comparison, a pattern is created which allows a clear assignment to the cycle stage. If this analysis is averaged over several hairs, a quantitative picture of the hair growth results, which can be referred to as a "molecular trichogram". Since a sample taken from humans as well as from other hairy mammals shows hair in the most diverse stages of development, mean values are determined for the expression levels of the individual genes. These mean values provide a reliable basis for the desired diagnosis.

If the pattern from the at least two genes deviates from the norm, the norm being a mammal without a hair growth disorder or without the same hair growth disorder or a sample or a molecular trichogram obtained from this sample, a hair growth disorder may be due to a pending, even phenotypic visible hair growth disorder can be closed. For the aforementioned deviation from the pattern, it is sufficient that a gene has a different expression. The method according to the invention is preferably validated in clinical studies. In the practical approach on which the invention is based, the expression of approximately 20,000 genes during the first and second hair cycle of the mouse (as an embodiment of the mammals which can be used in the method according to the invention) was measured by means of DNA chips and the expression levels were analyzed bioinformatically in such a way that groups of equally regulated genes were formed. The expression course of these groups was then compared with the individual phases of the hair cycle and those genes were defined which are characteristic of individual phases of the hair cycle. This procedure is explained in more detail in Example 1, which also shows that characteristic genes from the individual phases can have defined expression sites in the hair follicle. The groups listed in Example 1 are, as is readily apparent to the person skilled in the art, arbitrarily divided. On the basis of his general specialist knowledge in combination with the teaching according to the invention presented in this description, the person skilled in the art could also come to other groupings. It is essential, however, that another grouping does not lead to another invention and that the essence of the invention is retained even in such a case.

If a hair growth disorder has been diagnosed with the method according to the invention, the treating physician can take preventive measures (e.g. therapy) which suppress the hair growth disorder or limit or even reverse the phenotypic expression.

The advantages of the method according to the invention over the methods known in the prior art are obvious. On the one hand, the method according to the invention is easier to standardize than the methods available in the prior art, since the molecular trichograms, which represent the comparison standards, can be called up via data carriers. Since the actual analysis can also be carried out on the computer after tissue preparation and molecular biological processing, the method according to the invention is both easier to carry out than the known methods and more reliable in diagnosis. In addition, the method according to the invention is significantly more meaningful than the methods known in the prior art. Advantageous is also that for a relevant diagnostic statement only the expression profiles of two different genes need to be analyzed.

The present invention will thus contribute to improving the molecular knowledge of the hair cycle, as a result of which new analytical methods could be developed or to create the basis for the development of further new analytical methods which all allow the condition and quality of the hair cycle in molecular terms Describe the detail of the mouse and human being precisely. These findings can help pharmacological development to find and develop drugs more efficiently. In addition to new diagnostic procedures, molecular analysis will also increase the efficiency of clinical trials.

Furthermore, the invention relates to a method for diagnosing the effectiveness of the treatment of a hair growth disorder in a mammal, wherein the expression of at least two cyclically expressed genes in a sample is analyzed and wherein (a) one of the genes has the maximum expression level during anagen, during the (b) second gene has the maximum level of expression during catagen or telogen.

The statements made regarding the first embodiment of the invention apply mutatis mutandis to this embodiment of the invention, with the exception that no diagnosis of a hair growth disorder is to be made here, but rather the effectiveness of the treatment of a hair growth disorder in a mammal is tested. In this embodiment in particular, the mammal is a human being, since cosmetic aspects also play an important role here.

The method according to the invention can be used to test whether treatment with a medicament, a medicinal product or another substance or a mixture of substances (all of which can be known in the prior art) has the desired effect. First of all, a molecular trichogram of the tissue or of hair with the hair growth disorder is optional created. A sample is then taken one or more times during the treatment and the gene expression of the genes mentioned is analyzed. If the gene expression pattern changes to the normal state of a tissue, the normal state being a tissue without hair growth disorder or, in a worsened embodiment, without this hair growth disorder, the effectiveness of the treatment can be assumed. In the event that the gene expression pattern does not change or does not change to the desired or necessary extent, the effectiveness of the treatment cannot be assumed, at least not to the desired extent.

A major advantage of this embodiment of the method according to the invention is that the gene expression patterns change much faster than the observable phenotype. In other words, the diagnosis does not have to be waited until one observes a change in hair growth itself (or not). Possible harmful side effects of the medication, medical device etc. are therefore reduced to a minimum if they are not effective. Another advantage is that you can use another drug after a relatively short period of time if it is ineffective or ineffective and test for effectiveness. This shortening of the test phases to the effectiveness of a medication etc. has obvious advantages. On the one hand, there is a chance that if a first drug is not effective, a drug can be found relatively quickly that has the desired effectiveness. This can stop the hair growth disorder at a relatively early stage and prevent irreversible damage. Losing head hair in particular has psychological disadvantages that should not be underestimated.

In a preferred embodiment of the method according to the invention, expression (c) of at least a third cyclically expressed gene is analyzed, this gene having the maximum expression level during anagens, during catagen or during telogen. This embodiment is particularly preferred because the analysis of the expression of a third gene increases the meaningfulness of the trichogram obtained and at the same time reduces the risk of incorrect conclusions. It is advantageous if the third gene has its expression maximum in the anagen, but in a different subphase than the gene mentioned under (a) above.

In a further preferred embodiment, the method according to the invention is designed in such a way that expression (d) of at least a fourth cyclically expressed gene is analyzed, this gene having the maximum expression level during anagens, during catagen or during telogen.

This preferred embodiment also harbors an even higher level of security in the diagnostic analysis. This also applies to all other embodiments described in the description below, which include the analysis of other genes.

In this embodiment, at least two genes are preferably analyzed which have their maximum expression during anagen. If two genes with expression maxima in the anagen are analyzed, it is further preferred that one of the two other genes has its expression maximum in the catagen and in the telogen. It is also preferred that three genes have their maximum expression during anagen. In this case it is preferred that the gene mentioned in (b) has its expression maximum in the telogen. In all of the above combinations, it is advantageous if two or more genes that have their expression maximum at the same stage (ie in the anagen, in the catagen or in the telogen) have different sub-stages (e.g. one gene in early anagen, the second in late anagen etc) occurs.

In another preferred embodiment of the method according to the invention, an expression (e) of at least a fifth cyclically expressed gene is analyzed, this gene having the maximum level of expression during anagen, during catagen or during telogen. It also applies to this embodiment that the expression maxima of the genes analyzed originate from as many different stages or sub-phases as possible. It is particularly preferred that one of the genes has its maximum expression in the telogen, a second has its maximum expression in the catagen and the other three genes have their maximum expression in different subphases of the anagen. This particularly preferred embodiment is also reflected in the three preferred embodiments shown below.

Thus, in an additionally preferred embodiment of the method according to the invention, at least one of the genes which have their maximum expression level in the anagen has this during the middle anagen.

A further preferred embodiment according to the invention relates to a method, wherein at least one of the genes which have their maximum expression level in anagen has this during early anagen.

In another preferred embodiment, an inventive one

The method shown is at least one of the genes that have its maximum

Have expression level in the anagen, this has anagen-catagen during the transition.

According to the method according to the invention it is further preferred that the genes listed under (a), (b), (c), (d) and (e) have representatives of different groups of cyclically expressed genes, the diversity of the groups being characterized in that that they have their expression maxima in different time periods (preferably different subphases) and that genes belonging to the same group have the same expression profiles.

The term “the same expression profiles” is not to be interpreted in such a way that the profiles must have an identical profile. Rather, it is sufficient that the expression profiles have a similar profile to that of the same group assigns. It is clear to the person skilled in the art that, in principle, the group assignment can also take place according to other standards. This can be done, for example, in such a way that the maximum or minimum permitted distances between the

Expression profiles of different genes can be changed compared to the classification used here without changing the group membership. For example, the parameters for belonging to a group can be classified in such a way that two, three, four, preferably five, but also six, seven, eight, nine, ten or even more groups are formed. However, it must be emphasized that a change in the group membership of individual or a large number of genes has no influence on the feasibility of the method according to the invention.

In a particularly preferred embodiment, the method according to the invention analyzes the expression of at least one further member from at least one and preferably from each group of the cyclically expressed genes.

In a further particularly preferred embodiment, the expression of at least two further members from at least one and preferably from each group of the cyclically expressed genes is analyzed.

In principle, various sources are conceivable for use in the method according to the invention. In another preferred embodiment of the method according to the invention, a sample is used which consists of skin or epilated hair. The epilated hair preferably comes from the scalp.

If the sample is a skin sample, it is particularly preferred that it consists of scalp.

This embodiment is also particularly preferred because a scalp sample (as well as hair epilated from the scalp) is the best and easiest way to draw conclusions about hair growth disorders such as alopecia. In another preferred embodiment of the method according to the invention, the mammal is a mouse.

The mouse is particularly suitable as a model system to collect experiences and values that can then be transferred to humans. With the mouse, it is particularly easy to analyze the various stages of hair development immediately after birth, as it is synchronized. An additional advantage is that the individual stages of hair development in the mouse and thus the overall cycle of a hair is / are relatively short.

A method according to the invention is also preferred, wherein the at least one gene (a) is selected from the group of genes which comprise the sequences represented by the gene bank deposit numbers listed in Table 1 or the human homologues therefor. All Genbank accession numbers listed in this application are available at http://www.ncbi.nlm.nih.gov/Genbank/index.html.

Alternatively, it is preferred that the gene (a) is selected from the group of genes which comprise the sequences represented by the gene bank deposit numbers listed in Tables 2 or 3 or the human homologues therefor.

The term "comprise" means that the genes are either identical to the sequences identified by the deposit numbers or that the sequences are (coding) parts of the genes, for example ESTs.

As is known in the prior art, data found from the mouse genes and their expression products can generally be transferred to humans or the corresponding human genes or expression products. In addition, due to the generally high degree of homology between mouse and human genes, the latter can be derived readily if the latter is known. This is also due to the results of the human genome project. In another preferred embodiment, the invention relates to a method in which the at least one gene (b) is selected from the group of genes which comprise the sequences represented by the gene bank deposit numbers listed in Table 4 or the human homologues therefor. Alternatively, it is preferred that gene (b) is selected from the group of genes which comprise the sequences represented by the gene bank deposit numbers listed in Table 5 or the human homologues therefor.

In another preferred embodiment, the invention relates to a method in which the at least one gene (c) is selected from the group of genes which comprise the sequences represented by the gene bank deposit numbers listed in Table 2 or the human homologues therefor. Alternatively, it is preferred that the gene (c) is selected from the group of genes comprising the sequences represented by the gene bank accession numbers listed in Tables 1, 3, 4 or 5 or the human homologues therefor. As with the preferred embodiments described above, it is preferred in the method according to the invention that as many genes as possible are examined which have their maximum expression in different phases or sub-phases. This also applies to the preferred embodiments described below. In this respect, for example, a gene from Table 5 is only preferred if, in the previous preferred embodiment, a gene from Table 4 was analyzed with regard to the level of expression.

A method is also preferred, wherein the at least one gene (d) is selected from the group of genes which comprise the sequences represented by the gene bank deposit numbers listed in Table 3 or the human homologues therefor.

As an alternative to this, it is preferred that the gene (d) is selected from the group of genes which the gene library listed in Tables 1, 2, 4 or 5 Accession numbers shown sequences or the human homologues include.

Also preferred is an embodiment of the method according to the invention, wherein the at least one gene (s) is selected from the group of genes which comprise the sequences represented by the gene bank deposit numbers listed in Table 5 or the human homologues therefor. Alternatively, it is preferred that the gene (a) is selected from the group of genes comprising the sequences represented by the gene bank accession numbers listed in Tables 1, 2, 3 or 4 or the human homologues therefor.

A method is also preferred, wherein the at least one gene (a) is the human homologue to the mouse gene mentioned with reference to Table 1.

A method is also preferred, wherein the at least one gene (b) is the human homologue to the mouse gene mentioned with reference to Table 4.

As an alternative to this and also preferred is a method, wherein the at least one gene (c) is the human homologue to the gene from the mouse mentioned with reference to Table 2.

In addition, a method is preferred, wherein the at least one gene (d) is the human homologue to the mouse gene mentioned with reference to Table 3.

In this series of preferred embodiments, a method is finally preferred, wherein the at least one gene (s) is the human homologue to the mouse gene referred to in Table 5. The invention further relates to a method for isolating an inhibitor of a hair growth disorder, wherein

(a) analyzing the expression of at least one of the genes listed in the preceding claims under (a) to (e) in a skin sample or in epilated hair which was taken from a mammal with a hair growth disorder;

(b) comparing the analysis result from (a) with the expression of the gene in a skin sample or in epilated hair taken from a mammal of the same species, which mammal does not have this or no hair growth disorder;

(c) the gene whose expression differs by at least a factor 3 in a mammal with hair growth disturbance compared to a mammal without this or without a hair growth disturbance in step (b), the mRNA transcribed therefrom or the polypeptide encoded thereby or a skin sample, in which this gene is expressed at an abnormal level of expression in contact with one or more test substances under conditions which allow the test substance to bind to the gene, the mRNA or the polypeptide or to other genes, mRNAs or polypeptides in the skin sample; and

(d) isolating the test substance (s) which leads to a reduction in the factor in the expression in step (b).

The method according to the invention is based on the principle that a deviation of the expression pattern of one of the genes listed above from the norm (see above) leads or can lead to a disturbance in hair growth and that a substance or a plurality of substances which this deviation from the Corrected norm, is an inhibitor of hair growth disorder.

According to the invention, it is established that the above-mentioned difference in expression makes up at least a factor of 3. Such a difference can be regarded as significant in the sense of the invention and promises to isolate effective inhibitors of hair growth disorders. It is noteworthy here that the method according to the invention both reduces and increases Expression compared to the norm can be detected. Accordingly, the inhibitors are to be regarded as activators or as expression inhibitors. With the method according to the invention it is also possible to isolate inhibitors or activators which act on the transcription as well as on the translation level.

If a skin sample in which the gene is abnormally expressed is analyzed with the method according to the invention, the effect of the test substance (s) on other genes or their expression products, which in turn have an influence on the expression of the gene, can also be examined. Such other genes can be, for example, regulatory genes or they can encode parts of a cascade in which the polypeptide encoded by the gene is also involved. Exposure of the test substance (s) to such other genes or their expression products can lead to a normalization of the abnormal expression of the gene. When analyzing a skin sample, care must be taken that the test substances become active intracellularly, i.e. can interact with the other genes etc. Methods that ensure this are known in the art.

According to the method according to the invention, it is particularly advantageous if the gene whose expression is listed in step (b) (the “reference gene”) comes from a mammal that has no hair growth disorder.

According to the invention it is further preferred that the mammal is a human. This is particularly the case because people of both sexes often have more or less serious mental health problems due to hair loss. Successful diagnosis or, even better, successful isolation of an effective inhibitor should therefore lead to an improvement in subjective well-being.

In the method according to the invention it is preferred that the expression of the gene is normalized compared to the expression of a housekeeping gene. This preferred embodiment of the invention is suitable for minimizing incorrect analyzes. The normalization against the expression level of a housekeeping gene is particularly suitable for enabling a relatively precise finding of the factor with which the abnormally expressed gene differs from the reference gene.

In another preferred embodiment of the method according to the invention, the hair growth disorder is an alopecia.

It is particularly preferred that the alopecia is Alopecia androgenetica.

Alternatively, it is particularly preferred that the hair growth disorder is hirsutism.

In a further preferred embodiment of the method according to the invention, the test substances or the gene is coupled to a readout system or a readout system is added to the test batch, the readout system after binding of the test substance (s) to the gene, the mRNA or (Poly) peptide provides a detectable signal.

In the method according to the invention it is further preferred that the test substances are low molecular weight substances, peptides, aptamers, antibodies or fragments or derivatives thereof.

The terms "fragments or derivatives" of antibodies are well known in the art. The fragments include Fab fragment, Fv fragments and F (ab ₂ ) 'fragments. The antibody derivatives include, for example, scFv fragments; cf. e.g. Harlow and Lane "Antibodies, A Laboratory Manual" CSH Press, Cold Spring Harbor 1988.

In particular, the peptides or low molecular weight substances can be isolated from commercially available substance libraries. It is also preferred according to the invention that the process is a high-throughput process.

High-throughput methods are particularly preferred because they accomplish the largest possible number of expression patterns to be examined with minimal expenditure of time. Advantageously, the samples to be analyzed are placed on solid supports such as e.g. Transfer microchips and fix them there according to generally known methods or also transfer them into microtiter plates, where in vitro assays can also be carried out at the level of expression.

It is particularly preferred that the method is computer-assisted.

In another preferred embodiment, the invention relates to a method, further comprising improving the pharmacological properties of the inhibitor identified by the method according to the invention, wherein

(a) the binding site of the inhibitor to the gene, the mRNA or the polypeptide and optionally the binding site of the gene, the mRNA or the polypeptide to the inhibitor are identified;

(b) modifying the binding site of the inhibitor and the gene, mRNA or polypeptide by molecular modeling; and

(c) modifying the inhibitor so that its binding specificity or binding affinity or binding avidity for the gene, the mRNA or the polypeptide is increased.

All methods used in the method according to the invention are conventional or can be easily derived from conventional methods by a person skilled in the art. For example, biological evidence based on the nature of the polypeptides described here can be used to determine the specificity or potency of the pharmaceuticals, with an increase in one or more Activities of the polypeptides can serve to monitor specificity or potency. Steps (1) and (2) can be carried out according to conventional protocols. A protocol for binding site-specific mutagenesis is described in Ling MM, Robinson BH. (1997) Anal. Biochem. 254: 157-178. The use of homology modeling in connection with binding site-specific mutagenesis for the analysis of structure-function relationships is described in Szklarz and Halpert (1997) Life Sei. 61: 2507-2520. Chimeric proteins are produced by ligation of the corresponding DNA fragments via a single restriction site using conventional cloning methods, as described in Sambrook, Fritsch, Maniatis "Molecular Cloning, a laboratory manual" (1989) Cold Spring Harbor Laboratory Press. A fusion of two DNA fragments, from which a chimeric DNA fragment encoding a chimeric protein is formed, can also be generated with the gateway system (life technologies), a system which is based on DNA fusion by recombination. An advantageous example of molecular modeling is the structure-based design of compositions that bind to HIV reverse transcriptase; this is described in Mao, Sudbeck, Venkatachalam and Uckun (2000). Biochem. Pharmacol. 60: 1251-1265.

The identification of the binding site of the drug by means of binding-specific mutagenesis and the analysis of chimeric proteins can take place, for example, by modifications in the primary (poly) peptide sequence which influence the affinity of the drug; this usually enables precise mapping of the binding pocket for the inhibitor or the drug constituted therefrom.

Regarding step (2), the following protocols can be used: Once the effector site for the inhibitor has been mapped, the exact residues that interact with different parts of the inhibitor can be identified using a combination of those from the mutagenesis studies and computer simulations of the Structure of the binding site information provided that the exact three-dimensional structure of the inhibitor / drug is known (if not, this can be calculated) Simulation can be calculated). If the inhibitor itself is a peptide, it can also be mutated to determine which residues interact with other residues in the desired polypeptide.

Finally, in step (3) the inhibitor can be modified to increase its binding affinity or its potency and specificity. For example, if there are electrostatic interactions between a particular residue of the desired polypeptide and a region of the inhibitor / drug molecule, the total charge in that region can be modified, thereby increasing this particular interaction.

Computer programs can be helpful in identifying binding sites. Suitable computer programs can thus be used to identify interactive sites of a putative inhibitor and the polypeptide by computer-assisted searching for complementary structure motifs (Fassina, Immunomethods 5 (1994), 114-120). Other suitable computer systems for the computer-aided design of proteins and peptides are described in the prior art, for example in Berry, Biochem. Soc. Trans. 22: 1033-1036 (1994); Wodak, Ann. NY Acad. Be. 1987, 501: 1-13; Pabo, Biochemistry 25 (1986), 5987-5991. Modifications of the (poly) peptide can be produced, for example, by peptidomimetics. Other inhibitors can also be identified by synthesizing combinatorial peptidomimetic libraries through successive chemical modification and testing of the compositions obtained. Methods for the production and use of combinatorial peptidomimetic libraries are described in the prior art, for example in Ostresh, Methods in Enzymology 267 (1996), 220-234 and Dorner, Bioorg. Med. Chem. 4 (1996), 709-715. In addition, the three-dimensional and / or crystallographic structure of the activators of the expression of the (poly) peptide according to the invention can be used for the design of peptidomimetic activators, for example in connection with the (poly) peptide identified according to the invention (Rose, Biochemistry 35 (1996), 12933 - 12944; Rutenber, Bioorg. Med. Chem. 4 (1996), 1545-1558). It is particularly preferred in the method according to the invention that the binding sites in step (a) are determined by site-specific mutagenesis.

A further preferred embodiment of the method according to the invention further comprises the improvement of the pharmacological properties of the inhibitor identified according to the method according to the invention or of the inhibitor improved according to the method according to the invention, the inhibitor being further modified as a lead structure to include a modified active center, a modified activity spectrum, a modified organ specificity, improved activity, reduced toxicity (an improved therapeutic index), reduced side effects, a delayed start of therapeutic activity or the length of therapeutic activity, changed pharmacokinetic parameters (absorption, distribution, metabolism or excretion), modified physicochemical parameters (Solubility, hygroscopic properties, color, taste, smell, stability, condition), improved general specificity, organ / tissue specific icity and / or an optimized form and route of administration, which by the esterification of carboxyl groups, hydroxyl groups with carboxylic acids, hydroxyl groups, for example, phosphates, pyrophosphates, sulfates, "hemisuccinates" or the formation of pharmaceutically acceptable salts, pharmaceutically acceptable complexes or the synthesis of pharmacologically active polymers or the introduction of hydrophilic groups, the introduction or exchange of substituents in aromatics or side chains, the change of the substituent pattern or the modification by the introduction of isosteric or bioisosteric groups or the synthesis of homologous compounds, or the introduction of branched side chains, the conversion of alkyl substituents to cyclic analogs, the derivatization of hydroxyl groups to ketals or acetals, the N-acetylation to amides, phenyl carbamates, the synthesis of Mannich bases or imines or by the conversion of ketones, aldehydes in ship bases, oximes, acetals, ketals, enol esters, oxaholidines, thiozolidines or combinations thereof. The various steps outlined above are well known. They involve or are based on quantitative analyzes of structure-action relationships (QSAR); see. Kubinyi, "Hausch-Analysis and Related Approaches", VCH Verlag, Weinheim 1992, as well as combinatorial (bio) chemistry, classical chemistry and other approaches; see e.g. Holzgrabe and Bechtold, Deutsche Apotheker Zeitung 140 (8) (2000), 813- 823rd

In another preferred embodiment of the method according to the invention, the identified or improved inhibitor is further pharmacologically improved by peptidomimetics.

Approaches based on peptidomimetics are known in the prior art and are described, for example, in Rose, Biochemistry 35 (1996), 12933-12944 or Rutenber, Bioorg. Med. Chem. 4 (1996), 1545-1558.

It is further preferred according to the invention that the inhibitor of hair growth disturbance is an activator of the transcription or translation of the gene, the mRNA or an activator of the activity of the polypeptide.

In another preferred embodiment of the method according to the invention, the inhibitor of hair growth disturbance is an inhibitor of the transcription or translation of the gene, the mRNA or an inhibitor of the activity of the polypeptide.

The two preferred embodiments described above have already been explained in more detail above.

In a further preferred embodiment, the invention relates to a method further comprising the production of a medicament or medical preparation, the inhibitor obtained or improved by the method according to the invention being formulated with a pharmaceutically acceptable carrier or diluent. Examples of suitable pharmaceutically acceptable carriers and / or diluents are known to the person skilled in the art and include, for example, phosphate-buffered saline solutions, water, emulsions such as, for example, oil / water emulsions, various types of wetting agents or detergents, sterile solutions, etc. Medicaments which comprise such carriers , can be formulated using known conventional methods. These drugs can be administered to an individual in an appropriate dose. Administration can be oral or parenteral, for example intravenous, intraperitoneal, subcutaneous, intramuscular, local, intranasal, intrabronchial, oral or intradermal, or via a catheter at one location in an artery. Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and organic ester compounds such as ethyl oleate, which are suitable for injections. Aqueous carriers include water, alcoholic aqueous solutions, emulsions, suspensions, saline solutions and buffered media. Parenteral carriers include sodium chloride solutions, Ringer's dextrose, dextrose and sodium chloride, Ringer's lactate, and bound oils. Intravenous carriers include, for example, liquid, nutrient and electrolyte supplements (such as those based on Ringer's dextrose). The drug may also include preservatives and other additives such as antimicrobial compounds, antioxidants, complexing agents and inert gases. Furthermore, depending on the intended specific use, other active ingredients such as interleukins, growth factors, differentiation factors, interferons, chemotactic proteins or a non-specific immunomodulatory agent can be included. The treating doctor determines the type of dosage according to the clinical factors. It is known to the person skilled in the art that the type of dosage depends on various factors, such as, for example, the body size or weight, the body surface, the age, gender or general health of the patient, but also on the agent to be administered specifically, the duration and mode of administration, and other medications that may be administered in parallel. A typical dose can be, for example, in a range between 0.001 and 1000 μg, with doses below or above this exemplary range, especially taking into account the factors mentioned above. In general, with regular administration of the composition according to the invention, the dose should be in a range between 1 μg and 10 mg units per day. The active substances in these preparations will usually be present in a concentration of greater than 10 μg / ml of a physiological buffer. However, they can also be present in solid form in a concentration of 0.1 to 99.5% by weight of the total mixture. In general, it has proven to be advantageous to use the active ingredient (s) in total amounts of about 0.001 to 100 mg / kg, preferably in total amounts of about 0.01 to 10 mg / kg body weight per 24 hours, optionally as a continuous infusion or in the form of several Single doses to be administered to achieve the desired result. If the composition is administered intravenously, the dose should be in the range between 1 μg and 10 mg units per kilogram of body weight per day. The drug can be administered topically, locally or systemically.

In addition, the invention relates to the use of an inhibitor identified by the method according to the invention for the manufacture of a medicament or medical device for the prevention or treatment of hair growth disorders.

Finally, the invention relates to a composition comprising one or more of the inhibitors which have been identified or improved by the method according to the invention.

The composition is preferably a pharmaceutical composition. In this embodiment, the composition contains a pharmaceutically acceptable carrier or diluent as described above.

Alternatively, the composition is a diagnostic composition. The individual components of the diagnostic composition are preferred packed in one or more containers. These can in turn be packed in packaging intended for sale, provided with appropriate instructions for using the diagnostic composition (the diagnostic kit).

The figures show:

Figure 1:

RNA in situ hybridization for two genes (AF020790 and X81581) during two stages of the hair cycle (anagen, catagen).

Figure 2:

Presentation of examined points in time according to the correspondence analysis. A coordinate in two-dimensional space is assigned to each point in time (= vector, the dimensions of which are described by the expression values for the genes examined at the given point in time).

Figure 3:

Representation of individual genes according to the correspondence analysis. A coordinate in two-dimensional space is assigned to each gene (= vector, the dimensions of which are defined by the expression values at the different times).

Figures 4-9:

Standardized expression profiles for selected gene groups, in single and combined representation.

If one considers the expression values for a gene over a complete cycle as frequencies, this suggests a standardization of the values to sum 1. As a result, the graphical representation of the gene expression is only determined by its time course, whereas the absolute expression values are disregarded. This makes it possible to compare the expression patterns of genes with widely differing absolute expression values. The tables AD form the basis for Figures 4-7. Standardized expression profiles are shown in the figures; for reasons of clarity, only the first 10 genes of a list (table) are shown.

Figures 10 and 11:

Representation of selected genes and the examined times according to the

Correspondence Analysis.

In both figures, only the genes listed in Tables A-D were used and the representation was limited to only 10 genes for reasons of clarity. Figure 11 combines Figures 2 and 10.

The examples illustrate the invention.

Example 1: Creation of a molecular trichogram in the mouse system

gene expression profile

Skin from the back of BALB / c mice of various postnatal ages, which was completely covered with hair, was isolated. A complete hair cycle was recorded from the first anagen to the second anagen. For each microarray hybridization, skin from 3 to 4 mice was collected, mRNA isolated and converted to labeled cRNA, which was hybridized with Mu19K microarrays which represent about 20,000 genes according to the manufacturer (Affymetrix). The Affymetrix microarray program was used for raw data analysis.

Statistical analysis of the data

When using the Affymetrix microarray program, some data sets were excluded that showed defined values for the expression of certain genes. Measurement were considered unreliable or uninteresting if that "Positive signal" was below 0.35, the absolute value was A (absence) or the difference value was NC (no change). For this reason, data records in which all data points meet one of these criteria were excluded from further analysis. where all difference values corresponded to NC from the statistical evaluation and an independent check .. Considering the fact that there is no certain lower limit for the average difference, this was arbitrarily set to 100 if it was below the mentioned value maximum and min as the minimum average difference in a data set, only those data sets were further analyzed in which the values met the following criteria: max> 500 and max / min> 5, max> 600 and max / min> 4, max> 700 and max / min> 3.5, the part of genes with which no reliable measurement was obtained was kept to a minimum. Using the criteria listed, 2173 genes with a significant change in expression level during the hair cycle were further analyzed. According to their maximum expression during the hair cycle (early anagen, medium anagen, late anagen / start of catagen, catagen and telogen), they were summarized / grouped in tables 1-5.

For further analysis, the interdependency of times and gene expression profiles was determined using the correspondence analysis (Greenacre, 1992). Starting with a gene of interest, genes with a similar expression pattern were determined using "jackknife" distances (Heyer et al., 1999). These analyzes are shown as examples in Example 2 and are intended to illustrate how more detailed information can be extracted from the data.

RT-PCR

To check the results of the microarray hybridizations, mRNA was isolated postnatally from the dorsal skin of BALB / c mice of different ages. After rewriting into cDNA, PCR amplifications were carried out using gene-specific primers. The amplification was done with 25 to 40 cycles performed to keep the products in a linear range and analyzed by separation on a 1.5% agarose gel.

In situ hybridization

To determine the gene expression profile in the skin, cross-sections of the back skin of BALB / c mice of different ages were made postnatally. Essentially, non-radioactive in-situ hybridizations were carried out as already described (Beul and Boehm, 2000). Both "sense" and "antisense" strands of gene-specific fragments were used as probes. These fragments were generated by PCR using gene-specific primers.

In Figure 1, the RNA in situ hybridization is shown for 2 genes with opposite expression course during the hair cycle.

Figure 1:

In situ hybridization to determine the expression locations of genes in the hair follicle.

Above: AF020790, a member of Group 3, is expressed in the pre-cortex of the hair follicle during anagen. No gene expression is detectable in Catagen and Telogen. The maximum expression is reached in the middle to late days. Bottom: No significant expression is detectable for X81581 as a representative of group 4 in the anagen. In catagen, during which the expression maximum is reached, the gene is active in the cells of the dermal papilla. AA000708 AB002664 AI506524 AI854638 AW122355 M98530

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AA197454 AA986986 AI047508 AI840476 AI854020 AW045203

AA197511 AB006960 AI047972 AI840579 AI854358 AW045471

AA198568 AB019577 AI117581 AI841689 AI854380 A 045575

AA200369 AB020063 AI117609 AI841771 AI854434 A 045915

AA209597 AB028921 AI117859 AI841858 AI854450 A 046335

AA212964 AB033921 AI118229 AI842065 AI854546 AW046392

AA214997 AB042523 AI118905 AI842277 AI854549 AW046458

AA216875 AB042524 AI155124 AI842351 AI854583 AW047228

AA220077 AB050554 AI155515 AI842675 AI854587 AW047323

AA231562 AF004941 AI155885 AI842760 AI854794 AW047688

AA237347 AF009605 AI155995 AI842936 AI854880 A 048291

AA272682 AF015811 AI181132 AI842968 AI854890 A 048495

AA289896 AF019564 AI194762 AI843106 AI876418 AW048581

AA387264 AF020308 AI226254 AI843308 AI882152 AW048779

AA420003 AF022110 AI226339 AI843604 AI987726 AW049105

AA462137 AF022432 AI227034 AI843911 AJ001418 A 049191

AA537653 AF026123 AI256150 AI844448 AJ005620 A 049623

AA541964 AF026799 AI314958 AI844797 AJ006033 AW049840

AA607672 AF030343 AI324874 AI844855 AJ010305 AW049906

AA611766 AF031467 AI326963 AI845124 AJ223855 AW049963

AA612365 AF035643 AI386279 AI845242 AJ242663 AW050017

AA615100 AF035645 AI426288 AI845672 AJ243895 A 050243

AA619470 AF037205 AI429766 AI845790 AJ243964 AW050271

AA623874 AF037370 AI447965 AI845798 AJ249706 A 060592

AA636171 AF038008 AI448688 AI846040 AJ250490 AW060885

AA655547 AF041054 AI461631 AI846185 AJ291751 AW061229

AA656449 AF041376 AI462012 AI846354 AJ299430 A 061234

AA667100 AF041859 AI462265 AI846517 AK002443 AW110940

AA672708 AF045887 AI503400 AI846672 AK002669 A 120711

AA673704 AF046908 AI504668 AI846721 AK003712 AW120868

AA674925 AF047838 AI506206 AI846934 AK004396 AW120874

AA690218 AF052453 AI506524 AI847054 AK005042 AW120981

AA693185 AF053235 AI527257 AI847141 AK005404 AW121052

AA693310 AF056194 AI591564 AI847177 AK007665 A 121069

AA710092 AF064748 AI614447 AI847230 AK008925 AW121217

AA718024 AF073294 AI626885 AI847396 AK009604 AW121244

AA718364 AF077738 AI643935 AI848045 AK010611 AW121389

AA726223 AF079565 AI644063 AI848050 AK010786 AW121635

AA726579 AF083497 AI746459 AI848663 AK011417 AW121680

AA726837 AF085745 AI747215 AI848783 AK011633 A 121733

AA735016 AF087434 AI786586 AI849035 AK014093 A 122076

AA760364 AF089751 AI789926 AI849498 AK014601 AW122083

AA763764 AF090686 AI790538 AI850441 AK014813 AW122430

AA764293 AF091390 AI834895 AI850638 AK021021 A 122478

AA771584 AF093669 AI835402 AI851282 AU022607 AW122839

AA789854 AF093775 AI835784 AI851375 AU041675 A 122897

AA791742 AF109905 AI836048 AI851416 AU042641 AW123810

AA797255 AF109906 AI836095 AI851671 AU067718 AW123955

AA797604 AF110520 AI836143 AI851830 AV010984 A 124225

AA832741 AF111346 AI838016 AI851893 AV027723 AW124386

AA833173 AF155353 AI838208 AI851901 AV035418 AW124470

AA839151 AF177032 AI838258 AI852144 AV063483 A 124841 AW124970 M11533 X13986

AW124975 M12571 X14961

AW125260 M17551 X15591

A 125343 M18228 X15592

A 125453 M21856 X16672

AW125478 M29394 X51397

AW125574 M29462 X52046

AW125815 M31680 X53526

AW125884 M34094 X53928

A 211861 M57626 X56824

AW212475 M58588 X57971

AW214142 M60803 X58251

A 215449 M62470 X59769

AW226939 M63245 X62622

AW226939 M63335 X62940

A 228316 M64292 X67140

AW244668 M74495 X69832

A 476599 M84324 X70764

A 487953 M96827 X70842

AW543271 U00674 X74616

AW551795 U02487 X76653

A 552423 U07235 X77952

A 553639 U09114 X78989

A 557747 U10374 X78990

A 763318 U12791 X81581

AW909756 U15012 X82648

A 911938 U15977 X83569

AW988998 U16959 X85983

AY033514 U17282 X93037

C85523 U18812 X95279

C87028 U18975 X98055

D00725 U20614 Y07688

D13139 U21110 Y07812

D29797 U22399 Y08486

D49473 U22516 Y12887

D50031 U22519 Y13090

D50411 U24181 Y14296

D63902 U26437 Y14334

D85923 U32395

D86419 U33005

D86420 U41465

D89728 U42443

J03928 U49430

J04953 U51335

L00653 U53218

L01062 U55060

L02331 U57999

L06039 U60473

L06047 U61362

L07063 U63146

L07645 U70139

L12030 U82758

L12447 U88624 23108 U89491 23769 W13016

L28095 15682 28835 58782

L29441 W62660

L33416 91688

L35599 X01026 42115 X04405

L57509 X05546

M10062 X07414 AI048424 AA833173 AI120389 AI845920 AV215898 A 259632

AA008591 AA840142 AI120960 AI847784 AV234080 AW259940

AA008600 AA867860 AI132554 AI848510 AV239662 A 476152

AA059835 AA870639 AI155192 AI849335 AV260484 A 496146

AA065629 AA880988 AI155242 AI849587 AV272622 AW552039

AA066623 AA881576 AI155549 AI850090 AV290217 A 556558

AA109094 AA915262 AI158945 AI850497 AV302187 A 743993

AA123396 AA915643 AI173274 AI851210 AV358386 AW910263

AA124795 AA931004 AI180900 AI851348 AV361403 D31788

AA138065 AA958476 AI181332 AI851858 AW011791 D45203

AA142387 AA959436 AI197339 AI851921 A 045201 D45850

AA144420 AA960657 AI266876 AI852006 A 045204 D76440

AA144469 AA980253 AI326621 AI852143 A 045546 D86232

AA145212 AB002664 AI326813 AI852199 A 047323 D89728

AA145919 AB007813 AI327072 AI852838 AW047581 D90146

AA162200 AB009615 AI386437 AI852849 AW048272 J00389

AA174516 AB009993 AI391327 AI852886 A 048585 J03484

AA175284 AB010144 AI414584 AI852989 AW048883 J03776

AA177653 AB012693 AI427403 AI853320 AW048915 J03877

AA183595 AB016587 AI427506 AI853387 AW049156 J04946

AA184363 AB017614 AI447446 AI853795 A 049373 J04992

AA217898 AB021860 AI462265 AI853798 AW049388 K02782

AA241774 AB024922 AI505355 AI854162 AW049506 L07803

AA250358 AB025198 AI506524 AI854866 A 049669 07924

AA260354 AB027566 AI526801 AI877261 AW049730 L17324

AA285779 AF000236 AI553451 AI882298 A 049860 L22550

AA289666 AF009513 AI591439 AI882309 A 049969 L27439

AA387637 AF010254 AI593508 AI882416 AW060645 L31397

AA389905 AF012916 AI593827 AI931692 A 060770 L48989

AA407176 AF017258 AI605579 AJ001633 AW061110 L49507

AA435436 AF019564 AI641819 AJ005621 AW108371 M10937

AA530125 AF022072 AI642389 AJ006474 A 120505 M12566

AA544955 AF024637 AI645484 AJ007376 AW121167 M12660

AA549533 AF026481 AI646750 AJ007972 A 121222 M13500

AA597090 AF029843 AI647933 AJ010108 A 121438 M14220

AA600647 AF030513 AI663687 AJ012754 AW121470 M15501

AA607672 AF038149 AI746846 AJ131395 AW121749 M17962

AA611111 AF040252 AI747215 AJ223208 A 121957 M17979

AA619807 AF049850 AI747654 AJ223361 A 122012 M18194

AA638815 AF055638 AI787183 AJ239082 A 122039 M18837

AA647048 AF059583 AI788798 AJ272504 AW122053 M21932

AA647405 AF064749 AI790592 AK003152 AW122101 M22479

AA655199 AF076156 AI834796 AK003194 AW122625 M27008

AA667270 AF077742 AI836034 AK003712 AW123051 M27034

AA670908 AF079528 AI836555 AK003906 AW123189 M29010

AA673260 AF084524 AI837737 AK004064 A 123372 M31419

AA675580 AF090334 AI837984 AK004670 AW123773 M31680

AA709944 AF090696 AI838503 AK006444 AW123857 M31775

AA710564 AF091390 AI838567 AK007458 A 124125 M32490

AA717826 AF093624 AI839436 AK008069 AW124961 M34896

AA727857 AF098633 AI840267 AK009092 AW124975 M35244

AA756506 AF099973 AI840283 AK009223 AW124988 M35247

AA759582 AF148216 AI840643 AK010169 AW125094 M36579

AA763276 AF155909 AI841689 AK010915 AW125253 M55561

AA764553 AF287746 AI841858 AK016715 AW125254 57683

AA789365 AF349573 AI841894 AK017447 AW125440 M57696

AA790442 AI006773 AI842031 AK018585 AW208678 M64086

AA790638 AI021707 AI842456 AK019499 AW210333 M69069

AA792425 AI035425 AI843572 AV056976 A 211637 M74123

AA792643 AI036047 AI843866 AV099920 AW215831 81086

AA796759 AI048265 AI844722 AV101646 A 228671 M98530

AA797286 AI115769 AI844749 AV134101 A 230891 U02313

AA823048 AI116109 AI845609 AV170775 AW231026 U03419 a

U10484 X67469 U13705 X69064 U15012 X73230 U15541 X78682 U16163 X84797 U18869 X89998 U19119 X90875 U20159 X97227 U20365 Y00500 U20949 Y07611 U21906 Y09257 U22324 Y10875 U25096 Y17808 U25844 Z11974 U27195 Z18272 U35741 Z49204 U43541 Z67748 U49430 Z80833 U51196 U59488 U68064 U72032 U72680 U73820 U76758 U86108 U87240 U87948 U88567 U89876 U93291 U96684 V00829 V00830 W08276 W12396 W82382 X00496 X03994 X04480 X06454 X12905 X13335 X13460 X14432 X14972 X15202 X16202 X52046 X52490 X52643 X53929 X56397 X58196 X5825562696 X27396 X58256269859 The sequence information listed in the tables above will continue to be available in its original form without restriction. The data can be accessed at Entrez (http://www.ncbi.nlm.nih.gov/Entrez/nucleotide/html) via the Sequence Revision History department. Tables 6-10 below explicitly state the version numbers (if any) of the data listed in Tables 1-5 valid for the priority date. However, these data can already be unambiguously derived for the person skilled in the art from the generally known path outlined in Tables 1-5.

AA000708 AB002664, .1 AI506524 AI854638 A 122355 M98530.1

AA002843 AB007599. .1 AI527642 AI877116 AW122433 U01915 .1

AA032310 AB009615. .1 AI550379 AI882309 AW122609 U02079 .1

AA050985 AB010833. .1 AI574275 AI891493 AW122632 U05837 .1

AA096556 AB012727. , 1 AI585945 AI893814 AW123269 U20159 .1

AA152809 AB020886. .1 AI594294 AI956194 AW123386 U21906 .1

AA154606 AB041587, .1 AI594717 AJ001261. .1 AW123514 U22324 .1

AA163960 AC002397. .1 AI596121 AJ002522. .1 AW123683 U44389 .1

AA170203 AF004666. .1 AI642389 AJ132616. .1 AW123695 U56819 .1

AA178128 AF006196. .1 AI648998 AJ132771. .2 AW124101 U57393 .1

AA183595 AF012710. .1 AI663687 AJ133533. , 1 AW124306 U60884, .1

AA200617 AF013119. .1 AI834796 AJ223087. , 1 AW124719 U72644. .1

AA204302 AF013486. , 1 AI834822 AJ223361. .1 A 124952 U76618, .1

AA240251 AF017111. , 1 AI835646 AJ245569. , 1 A 125 170 U86405, .1

AA267422 AF024637. , 1 AI835703 AK002240. , 1 A 125284 U93583, .1

AA269694 AF029791. , 1 AI835826 AK003194. .1 AW125710 V00817. .1

AA277664 AF029843. , 1 AI836721 AK003524. 1 A 125757 V00821. .1

AA388082 AF038008. .1 AI838112 AK003838. 1 AW209229 X00496. .1

AA433547 AF064749. .1 AI838320 AK005293. 1 AW210333 X04017. .1

AA444309 AF076156. 1 AI838737 AK006444. 1 A 212915 X06368. .1

AA444797 AF083032. .1 AI839232 AK018686. 1 A 214225 X13333. .1

AA500406 AF084524. 1 AI839841 AK019964. 1 A 214617 X16426. .1

AA512730 AF090334. 1 AI840035 AV027479 A 215808 X52490. .1

AA546705 AF132744. 1 AI840437 AV028168 A 228933 X53247. .1

AA560171 AF143955. 1 AI840577 AV033602 AW491689 X54401. .1

AA592182 AF237917. 2 AI841087 AV076672 AW555641 X57687. ,1

AA615200 AF262985. , 1 AI842612 AV117844 D13543.1 X58609. .1

AA615624 AI019999 AI843031 AV160578 D37837.1 X60671. .1

AA617489 AI020480 AI843040 AV211554 D44456.1 X62154. .1

AA624041 AI021125 AI843182 AV228966 D50031.1 X67668. .1

AA647303 AI049139 AI843574 AV230737 D86726.1 X67809. .1

AA647507 AI049398 AI843864 AV231601 D88791.1 X76066. .1

AA656550 AI060627 AI844626 AV263513 J03298.1 X78936. .1

AA666504 AI115746 AI844864 AV331859 J04992.1 X78990. .1

AA672634 AI116463 AI845380 AV334573 J05277.1 X87128. .1

AA681764 AI117666 AI845988 AW045453 L12447.1 X94915. .1

AA682063 AI120758 AI846341 A 045665 L20315.1 Y00225. .1

AA691890 AI121796 AI846739 AW046072 L20343.1 Y09864. .1

AA693175 AI121807 AI847177 AW046136 L22550.1 Z24722. ,1

AA718842 AI131895 AI847230 A 046748 L35032.1 Z31689. ,1

AA727413 AI132585 AI847267 AW047156 M10937.1 Z54209. ,1

AA733594 AI152741 AI847483 AW047224 M12347.1 Z72486. ,1

AA733937 AI154017 AI848448 AW047331 M12660.1

AA756724 AI154249 AI848671 A 047744 M13500.1

AA764332 AI154452 AI848835 A 048601 M13963.1

AA790307 AI156274 AI848915 A 048860 M15501.1

AA790898 AI173274 AI849532 AW049181 M18837.1

AA791012 AI196645 AI851309 AW049504 M19279.1

AA792425 AI314784 AI851574 A 050335 M21285.1

AA792643 AI315312 AI851714 A 060556 M22531.1

AA823901 AI317205 AI851770 A 061324 M26270.1

AA869027 AI326984 AI851968 A 107248 M27034.1

AA882422 AI414973 AI852086 AW107509 M29395.1

AA915460 AI426284 AI852387 AW107813 M31312.1

AA919924 AI426288 AI852553 A 108371 M32489.1

AA920123 AI426400 AI852641 AW120796 M34896.1

AA921463 AI450714 AI852672 AW120882 M35986.1

AA929330 AI452117 AI852679 AW121237 M64360.1

AA930519 AI466326 AI852777 AW121291 M64817.1

AA959852 AI466577 AI853127 A 121823 M69069.1

AA967585 AI481642 AI853303 AW122024 M76987.1

AA981032 AI505194 AI853685 AW122038 M77174.1

AA981342 AI505484 AI853924 AW122077 M91380.1 AA007891 AA764384 AI120142 AI842846 AJ009634.1 AW049504

AA015575 AA764577 AI121731 AI842969 AJ010984 .1 AW049684

AA017860 AA790833 AI122538 AI843178 AJ223293 .1 A 049829

AA030475 AA794499 AI152262 AI843686 AJ223834. .1 A 050102

AA032456 AA795285 AI153421 AI843874 AJ237847, .1 AW050141

AA048058 AA795610 AI153647 AI843880 AJ251685, .1 AW050325

AA061949 AA795830 AI155725 AI844864 AK003838. .1 A 060443

AA073823 AA796207 AI156452 AI845303 AK004697, .1 AW060657

AA098047 AA796831 AI157167 AI845529 AK004716. .1 A 060674

AA119793 AA797317 AI157666 AI846017 AK004996. .1 AW061016

AA120064 AA798365 AI173038 AI846126 AK005038. .1 A 061179

AA124346 AA798624 AI182073 AI846206 AK010876. .1 AW061267

AA139852 AA798971 AI183023 AI846531 AK011277. .1 A 061276

AA144272 AA799269 AI197311 AI846766 AK014626. .1 A 061324

AA146383 AA816240 AI226158 AI846792 AK014776. .1 A 107218

AA170647 AA821390 AI255271 AI847001 AK016716. .1 A 120568

AA183037 AA822187 AI322971 AI847236 AK018368. .1 AW120614

AA189403 AA833077 AI324871 AI847526 AK018510. .1 AW120623

AA199023 AA833293 AI326889 AI847915 AU017342 A 120751

AA199122 AA838903 AI385977 AI848386 AV008737 AW120804

AA221081 AA839061 AI413858 AI848479 AV009583 A 120867

AA261707 AA839379 AI427138 AI848574 AV028139 AW120906

AA267124 AA863768 AI452103 AI848732 AV075775 A 120926

AA275367 AA874496 AI461704 AI848813 AV076550 AW120954

AA396604 AA913994 AI462170 AI848982 AV083230 AW121110

AA416364 AA914839 AI462491 AI849082 AV087476 A 121249

AA433654 AA967263 AI464230 AI849135 AV108842 A 121433

AA444896 AA990118 AI505400 AI849333 AV119513 AW121498

AA462886 AB005132. , 1 AI551087 AI849354 AV167174 AW121676

AA473599 AB013607. .1 AI573468 AI849439 AV216992 AW121724

AA475175 AB013819. , 1 AI593501 AI849676 AV228145 AW121740

AA517145 AB013849. .1 AI594290 AI849718 AV228287 AW122058

AA543464 AB013912. , 1 AI596020 AI849958 AV229826 A 122189

AA544721 AB017189. , 1 AI605905 AI850362 AV292847 AW122271

AA546366 AB021961. , 1 AI606257 AI850666 AV292940 AW122284

AA591295 AB035511. , 1 AI614117 AI850846 AV294837 AW122331

AA600555 AB045313. , 1 AI645662 AI851198 AV306274 AW122472

AA624068 AF007802. .1 AI646302 AI851230 AV334961 AW122703

AA637016 AF010133. .1 AI663783 AI851583 AV339309 AW122706

AA646434 AF018262. .1 AI787627 AI851727 AW012633 AW122863

AA655369 AF022465. 1 AI790931 AI851762 A 045261 A 122882

AA671429 AF030001. 1 AI835864 AI851983 AW045958 A 122949

AA671515 AF042822. 3 AI836451 AI852017 AW045975 A 123120

AA672796 AF050165. 1 AI836780 AI852022 A 045981 A 123135

AA673500 AF053232. 1 AI836870 AI852273 A 046003 AW123237

AA681520 AF053454. 1 AI837254 AI852645 AW046039 A 123267

AA681520 AF055889. 1 AI838052 AI852770 A 046381 AW123273

AA683785 AF058054. 1 AI838080 AI852880 A 046443 AW123283

AA683840 AF070970. 1 AI838704 AI852988 AW046563 AW123297

AA684073 AF107296. 1 AI838772 AI853294 A 046889 A 123461

AA688930 AF109906. 1 AI839366 AI853644 AW047525 A 123785

AA691668 AF113005. 1 AI839647 AI854144 AW047866 A 124023

AA693246 AF134346. 2 AI840320 AI854287 AW047915 AW124049

AA693298 AF291655. 1 AI840336 AI854445 A 048159 AW124340

AA710132 AF317517. 1 AI840436 AI854606 A 048174 AW124483

AA711620 AF326960. 2 AI840577 AI854767 AW048282 AW124520

AA716925 AI005782 AI841088 AI876370 AW048685 A 124544

AA717560 AI006190 AI841093 AI891475 AW048954 AW124680

AA727943 AI047601 AI841279 AI891742 AW049111 A 125103

AA760211 AI049099 AI841497 AJ001260.1 AW049226 AW125510

AA760414 AI049103 AI841645 AJ002306.1 AW049260 AW125713

AA763244 AI060627 AI841654 AJ003132.1 AW049420 AW146312

AA763275 AI115629 AI842259 AJ007909.1 A 049487 AW209158 AW209238 U63648..1

A 213883 U66249. ,1

A 214124 U69135. .1

AW215736 U76374. .2

AW227345 U80932. .1

A 229312 U83902. ,1

AW230369 U85414. .1

A 318984 V00722. .1

AW542832 37034

A 546010 W39793

A 557814 W71352

AW908136 W77210

A 989451 X07414. .1

D12644.1 X12944. .1

D13545.1 X14309. .1

D16503.1 X14678. .1

D26089.1 X14759. .1

D26091.1 X15267, .1

D26107.1 X56304. .1

D26483.1 X57377, .1

D55720.1 X60676. .1

D83966.1 X61399, .1

D85612.1 X63349 .1

D87908.1 X65553 .1

D89664.1 X75483. .1

J04620.1 X75649 .1 21027.1 X75650 .1 28836.1 X77602 .1

L32836.1 X77731 .1

L39017.1 X94478 .1

M12056.1 X94694 .1

M13805.1 X94915 .1

M16356.1 Y00864 .1

M16358.1 Y11245 .1

M16359.1 Y15128 .1

M16360.1 Y15197 .1

M17818.1 Z80941 .1

M28730.1

M29260.1

M31690.1

M32032.1

M33988.1

M35153.1

M35970.1

M62766.1

M63709.1

M73818.1

M77831.1

M80206.1

M83749.1

M98036.1

M99167.1

U01915.1

U03711.1

U11274.1

U12620.1

U27177.1

U28789.1

U32446.1

U39074.1

U43918.1

U44955.1

U51805.1 AA013838 AB018002.1 AI595652 AI853265 AW060996 M30641.1

AA013990 AB022053 .1 AI605409 AI853561 A 061306 M31775 .1

AA014220 AB024566 .1 AI606234 AI853562 A 120734 M63695 .1

AA015501 AB026432, .1 AI614021 AI853644 AW120756 M63697 .1

AA016713 AB028241, .1 AI626942 AI854510 AW120786 M64292 .1

AA033036 AB028272, .1 AI642445 AI854638 AW120925 M70642 .1

AA058258 AB033744. .1 AI642522 AI854835 AW121110 M81445 .1

AA107834 AB041594. .1 AI644019 AI882080 A 121606 M92088 .1

AA111647 AB042524. .1 AI644158 AI892704 AW121759 M92420, .1

AA116391 AB053477. .1 AI645662 AJ006972. , 1 AW121865 U01063, .1

AA118877 AF003525. .1 AI648981 AJ131526. .1 A 122075 U02880 .1

AA142814 AF003867. .1 AI663783 AJ249901. .1 AW122093 U08215 .1

AA153346 AF004100. .1 AI790909 AJ251685. .1 AW122476 U08378. .1

AA166148 AF006602. .2 AI835482 AK003169. .1 A 122985 U09268. .1

AA189793 AF016482. .1 AI835750 AK004699. , 1 AW123286 U09507. .1

AA198542 AF017994. .1 AI835817 AK004782. , 1 AW123345 U13705, .1

AA210246 AF020790. .1 AI837322 AK005344. .1 A 123347 U16322. .1

AA222909 AF021836. .1 AI838149 AK008561. .1 A 123363 U19520. .1

AA254003 AF032714. , 1 AI838622 AK009313. 1 AW123563 U20611. .1

AA267450 AF033115. , 1 AI839150 AK009744. 1 A 124072 U20735. .1

AA273350 AF033565. .1 AI839225 AK010275. .1 AW124311 U35312, .1

AA276035 AF034745. .1 AI839742 AK010371. , 1 A 124369 U35374, .1

AA415611 AF039391. .1 AI840376 AK010382. .1 A 124387 U41341. .1

AA458138 AF041060. .1 AI840484 AK010772. .1 AW124812 U49507. .1

AA467442 AF049850. , 1 AI841112 AK010860. 1 A 124857 U60150. .1

AA498900 AF079834. .1 AI841137 AK013932. 1 AW124910 U66249. .1

AA500406 AF087687. , 1 AI841945 AK014635. 1 AW125308 U73170. .1

AA530761 AF113005. .1 AI842428 AK015667. 1 A 125843 U83148. .1

AA544895 AF184973. 1 AI843176 AK017569. 1 A 208678 U90662. .1

AA590959 AF213382. .1 AI843520 AK019830. 1 AW210542 U95826. .1

AA607761 AF218809. 1 AI844151 AU020383 AW212851 W36621

AA612146 AF231120. , 1 AI844179 AU021685 A 213683 W41539

AA646801 AF256218. , 1 AI844719 AU051782 AW214532 W49316

AA647393 AF256219. , 1 AI844839 AU079846 AW215449 70956

AA656805 AF274866. , 1 AI845321 AU079986 A 227996 W81894

AA672796 AF286724. .1 AI845579 AV087613 A 228014 84231

AA688930 AF378830. .1 AI845634 AV093019 A 321422 X03233. .1

AA717842 AI006348 AI845953 AV218363 AW412499 X06086. .1

AA718169 AI019722 AI846729 AV222614 AW490358 X07699. ,1

AA727104 AI019931 AI846984 AV233152 A 490923 X13945. ,1

AA727424 AI020117 AI847060 AV236307 A 492472 X14678. .1

AA733734 AI020377 AI847236 AV238964 A 543347 X16995. .1

AA738853 AI036466 AI847528 AV312105 AW545194 X53928. .1

AA739364 AI036798 AI848032 AV338003 AW556386 X55123. .1

AA755053 AI050252 AI848107 AV354964 AW557254 X59252. .1

AA763275 AI060595 AI848392 AV367683 AW825484 X71978. ,1

AA763888 AI132203 AI848623 AV373702 D10727.1 X83733. 2

AA789879 AI152867 AI848986 AW045599 D14077.1 X94478. ,1

AA791234 AI155095 AI849070 AW045842 D16141.1 X99143. .1

AA791965 AI155297 AI849082 A 045920 D16432.1 Y00094. 1

AA793666 AI156772 AI849131 A 046322 D86424.1 Y07915. 1

AA798461 AI157260 AI849601 AW046452 00919.1 Y08460. 1

AA798859 AI173190 AI851057 AW047109 12029.1 Y14696. .1

AA799222 AI180528 AI851230 AW047756 L35599.1 Z22532. .1

AA832685 AI255982 AI851272 AW048228 M13805.1 Z47766. .1

AA839237 AI256314 AI851561 A 048468 M17878.1 Z67963. 1

AA871648 AI325614 AI851600 AW048625 M19436.1

AA874620 AI429433 AI851614 AW048758 M25944.1

AA881294 AI447780 AI852349 AW049824 M26270.1

AB006463.1 AI466034 AI852461 AW049974 M26385.1

AB012033.1 AI573804 AI852624 AW049986 M27266.1

AB012725.1 AI592005 AI852805 AW050015 27695.1

AB013607.1 AI592996 AI853243 AW060536 M27734.1 AI852003 AA840209 AF179369.1 AI838997 AI852237 AV091103

AA048213 AA879591 AF180471. 1 AI839335 AI852255 AV216479

AA050582 AA879929 AF246265. 1 AI839531 AI852358 AV233109

AA058307 AA881340 AF277718. 1 AI839544 AI852519 AV234777

AA106751 AA882285 AF297860. 1 AI839688 AI852653 AV243890

AA108501 AA919594 AF303450. 1 AI839697 AI852667 AV267248

AA137485 AA939571 AF312007. 1 AI839746 AI852672 AV318082

AA138087 AA981257 AI007127 AI840158 AI852911 AV346092

AA138736 AA981778 AI007160 AI840339 AI853095 AV364118

AA153346 AA985765 AI021069 AI840348 AI853900 A 011791

AA163792 AA986141 AI035547 AI840446 AI853918 A 012588

AA197454 AA986986 AI047508 AI840476 AI854020 AW045203

AA197511 AB006960 .1 AI047972 AI840579 AI854358 AW045471

AA198568 AB019577. .1 AI117581 AI841689 AI854380 A 045575

AA200369 AB020063. .1 AI117609 AI841771 AI854434 AW045915

AA209597 AB028921. .1 AI117859 AI841858 AI854450 AW046335

AA212964 AB033921. .1 AI118229 AI842065 AI854546 AW046392

AA214997 AB042523, .1 AI118905 AI842277 AI854549 AW046458

AA216875 AB042524. .1 AI155124 AI842351 AI854583 AW047228

AA220077 AB050554, .1 AI155515 AI842675 AI854587 AW047323

AA231562 AF004941. .1 AI155885 AI842760 AI854794 AW047688

AA237347 AF009605. .1 AI155995 AI842936 AI854880 A 048291

AA272682 AF015811, .1 AI181132 AI842968 AI854890 A 048495

AA289896 AF019564. .1 AI194762 AI843106 AI876418 AW048581

AA387264 AF020308. .1 AI226254 AI843308 AI882152 AW048779

AA420003 AF022110. .1 AI226339 AI843604 AI987726 A 049105

AA462137 AF022432, .1 AI227034 AI843911 AJ001418 AW049191

AA537653 AF026123, .1 AI256150 AI844448 AJ005620 AW049623

AA541964 AF026799. .1 AI314958 AI844797 AJ006033 AW049840

AA607672 AF030343. .1 AI324874 AI844855 AJ010305 A 049906

AA611766 AF031467, .1 AI326963 AI845124 AJ223855 AW049963

AA612365 AF035643. .1 AI386279 AI845242 AJ242663.1 AW050017

AA615100 AF035645, .1 AI426288 AI845672 AJ243895.1 AW050243

AA619470 AF037205. .1 AI429766 AI845790 AJ243964 AW050271

AA623874 AF037370. .1 AI447965 AI845798 AJ249706 AW060592

AA636171 AF038008. .1 AI448688 AI846040 AJ250490 AW060885

AA655547 AF041054. .1 AI461631 AI846185 AJ291751 A 061229

AA656449 AF041376. .1 AI462012 AI846354 AJ299430 A 061234

AA667100 AF041859. .1 AI462265 AI846517 AK002443 AW110940

AA672708 AF045887. .1 AI503400 AI846672 AK002669 A 120711

AA673704 AF046908. .1 AI504668 AI846721 AK003712 AW120868

AA674925 AF047838. .1 AI506206 AI846934 AK004396 AW120874

AA690218 AF052453. .1 AI506524 AI847054 AK005042 AW120981

AA693185 AF053235. , 1 AI527257 AI847141 AK005404 AW121052

AA693310 AF056194. , 1 AI591564 AI847177 AK007665.1 A 121069

AA710092 AF064748. , 1 AI614447 AI847230 AK008925.1 AW121217

AA718024 AF073294. , 1 AI626885 AI847396 AK009604.1 A 121244

AA718364 AF077738. .1 AI643935 AI848045 AK010611.1 A 121389

AA726223 AF079565. .1 AI644063 AI848050 AK010786 AW121635

AA726579 AF083497. .1 AI746459 AI848663 AK011417 AW121680

AA726837 AF085745. .1 AI747215 AI848783 AK011633 AW121733

AA735016 AF087434. .1 AI786586 AI849035 AK014093 A 122076

AA760364 AF089751. .1 AI789926 AI849498 AK014601 A 122083

AA763764 AF090686. .1 AI790538 AI850441 AK014813 AW122430

AA764293 AF091390. , 1 AI834895 AI850638 AK021021.1 AW122478

AA771584 AF093669. , 1 AI835402 AI851282 AU022607 AW122839

AA789854 AF093775. , 1 AI835784 AI851375 AU041675 AW122897

AA791742 AF109905. .1 AI836048 AI851416 AU042641 AW123810

AA797255 AF109906. .1 AI836095 AI851671 AU067718 AW123955

AA797604 AF110520. .1 AI836143 AI851830 AV010984 AW124225

AA832741 AF111346. .1 AI838016 AI851893 AV027723 A 124386

AA833173 AF155353. .1 AI838208 AI851901 AV035418 AW124470

AA839151 AF177032. 1 AI838258 AI852144 AV063483 A 124841 AW124970 M11533..1 X13986.1

A 124975 M12571, .1 X14961.1

AW125260 M17551. .1 X15591.1

A 125343 M18228. .1 X15592.1

A 125453 M21856. , 1 X16672.1

AW125478 M29394. .1 X51397.1

AW125574 M29462. .1 X52046

AW125815 M31680. .1 X53526

AW125884 M34094, .1 X53928

AW211861 M57626. .1 X56824

AW212475 M58588. , 1 X57971.1

AW214142 M60803. , 1 X58251.1

AW215449 M62470. .1 X59769.1

AW226939 M63245. .1 X62622.1

A 226939 M63335. .1 X62940.1

AW228316 M64292. .1 X67140.1

AW244668 M74495. , 1 X69832.1

AW476599 M84324. .1 X70764.1

AW487953 M96827. , 1 X70842.1

A 543271 U00674. .1 X74616

AW551795 U02487, .1 X76653.

AW552423 U07235. .1 X77952.

AW553639 U09114. .1 X78989.

AW557747 U10374. .1 X78990.

A 763318 U12791. .1 X81581.1

AW909756 U15012. .1 X82648.1

AW911938 U15977, .1 X83569.1

AW988998 U16959. .1 X85983.1

AY033514.1 U17282. .1 X93037.1

C85523 U18812. .1 X95279.1

C87028 U18975. .1 X98055.1

D00725.1 U20614. .1 Y07688.1

D13139.1 U21110. .2 Y07812.1

D29797.1 U22399. .1 Y08486.1

D49473.1 U22516, .1 Y12887.1

D50031.1 U22519, .1 Y13090.1

D50411.1 U24181. .1 Y14296.1

D63902.1 U26437, .1 Y14334.1

D85923.1 U32395. .1

D86419.1 U33005. .1

D86420.1 U41465. .1

D89728.1 U42443. .1

J03928.1 U49430. .1

J04953.1 U51335. .1

L00653.1 U53218. .1

L01062.1 U55060. .1

L02331.1 U57999. .1

L06039.1 U60473. .1 06047.1 U61362. .1

L07063.1 U63146, .1 07645.1 U70139. .1

L12030.1 U82758. .1

L12447.1 U88624. 2

L23108.1 U89491. .1 23769.1 W13016 28095.1 15682

L28835.1 58782

L29441.1 W62660

L33416.1 91688

L35599.1 X01026. ,1

L42115.1 X04405. .1

L57509.1 X05546. .1 10062.1 X07414. .1 AI048424 AA833173 AI120389 AI845920 AV215898 AW259632

AA008591 AA840142 AI120960 AI847784 AV234080 AW259940

AA008600 AA867860 AI132554 AI848510 AV239662 A 476152

AA059835 AA870639 AI155192 AI849335 AV260484 A 496146

AA065629 AA880988 AI155242 AI849587 AV272622 AW552039

AA066623 AA881576 AI155549 AI850090 AV290217 AW556558

AA109094 AA915262 AI158945 AI850497 AV302187 A 743993

AA123396 AA915643 AI173274 AI851210 AV358386 A 910263

AA124795 AA931004 AI180900 AI851348 AV361403 D31788 .1

AA138065 AA958476 AI181332 AI851858 AW011791 D45203 .1

AA142387 AA959436 AI197339 AI851921 A 045201 D45850 .1

AA144420 AA960657 AI266876 AI852006 AW045204 D76440 .1

AA144469 AA980253 AI326621 AI852143 AW045546 D86232 .1

AA145212 AB002664, .1 AI326813 AI852199 A 047323 D89728 .1

AA145919 AB007813, .1 AI327072 AI852838 AW047581 D90146 .1

AA162200 AB009615, .1 AI386437 AI852849 A 048272 J00389 .1

AA174516 AB009993. .1 AI391327 AI852886 A 048585 J03484 .1

AA175284 AB010144, .1 AI414584 AI852989 A 048883 J03776 .1

AA177653 AB012693. .1 AI427403 AI853320 A 048915 J03877 .1

AA183595 AB016587. .1 AI427506 AI853387 AW049156 J04946 .1

AA184363 AB017614. .1 AI447446 AI853795 AW049373 J04992. .1

AA217898 AB021860. .1 AI462265 AI853798 A 049388 K02782, .1

AA241774 AB024922. .1 AI505355 AI854162 AW049506 07803, .1

AA250358 AB025198. .1 AI506524 AI854866 A 049669 07924, .1

AA260354 AB027566. .1 AI526801 AI877261 A 049730 L17324, .1

AA285779 AF000236. .1 AI553451 AI882298 AW049860 22550. .1

AA289666 AF009513, .1 AI591439 AI882309 AW049969 27439 .1

AA387637 AF010254. .1 AI593508 AI882416 AW060645 L31397. .1

AA389905 AF012916. .1 AI593827 AI931692 AW060770 L48989. .1

AA407176 AF017258. .1 AI605579 AJ001633. .1 A 061110 L49507. .1

AA435436 AF019564. .1 AI641819 AJ005621. , 1 A 108371 M10937. .1

AA530125 AF022072. .1 AI642389 AJ006474. , 1 AW120505 M12566, .1

AA544955 AF024637. .1 AI645484 AJ007376. .1 A 121167 M12660, .1

AA549533 AF026481. .1 AI646750 AJ007972. .1 AW121222 M13500, .1

AA597090 AF029843. .1 AI647933 AJ010108. , 1 A 121438 M14220, .1

AA600647 AF030513. .1 AI663687 AJ012754. .1 AW121470 M15501. .1

AA607672 AF038149. .1 AI746846 AJ131395. .1 AW121749 M17962. .1

AA611111 AF040252. .1 AI747215 AJ223208. .1 A 121957 M17979. .1

AA619807 AF049850. .1 AI747654 AJ223361. 1 AW122012 M18194. .1

AA638815 AF055638. .1 AI787183 AJ239082. 1 AW122039 M18837. .1

AA647048 AF059583. , 1 AI788798 AJ272504. 1 AW122053 M21932. .1

AA647405 AF064749. .1 AI790592 AK003152. 1 A 122101 M22479. .1

AA655199 AF076156. .1 AI834796 AK003194. 1 A 122625 M27008. .1

AA667270 AF077742. , 1 AI836034 AK003712. 1 AW123051 M27034. .1

AA670908 AF079528. , 1 AI836555 AK003906. 1 A 123189 M29010. .1

AA673260 AF084524. .1 AI837737 AK004064. 1A 123372 M31419. ,1

AA675580 AF090334. .1 AI837984 AK004670. 1 AW123773 M31680. ,1

AA709944 AF090696. .1 AI838503 AK006444. 1A 123857 M31775. .1

AA710564 AF091390. , 1 AI838567 AK007458. 1 A 124125 M32490. .1

AA717826 AF093624. , 1 AI839436 AK008069. 1 A 124961 M34896. .1

AA727857 AF098633. , 1 AI840267 AK009092. 1 A 124975 M35244. .1

AA756506 AF099973. , 1 AI840283 AK009223. 1 AW124988 M35247. .1

AA759582 AF148216. , 1 AI840643 AK010169. 1 AW125094 M36579. .1

AA763276 AF155909. .1 AI841689 AK010915. 1 AW125253 M55561. .1

AA764553 AF287746. .1 AI841858 AK016715. 1 A 125254 M57683. ,1

AA789365 AF349573. .1 AI841894 AK017447. 1 AW125440 M57696. ,1

AA790442 AI006773 AI842031 AK018585. 1 A 208678 M64086. ,1

AA790638 AI021707 AI842456 AK019499. 1 AW210333 M69069. .1

AA792425 AI035425 AI843572 AV056976 AW211637 M74123. ,1

AA792643 AI036047 AI843866 AV099920 AW215831 M81086. .1

AA796759 AI048265 AI844722 AV101646 A 228671 M98530. .1

AA797286 AI115769 AI844749 AV134101 A 230891 U02313. .1

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X62622. Example 2: Evaluation of the data on the hair cycle

1. Raw data

Expression of approx. 20,000 genes at 9 points in time: 9, 15, 17, 19, 20, 25, 24, 25, 27 and 31.

Gen g and times / ^' :

Positive fraction: Pos {g, /)

Absolute Call: AbsCall (flf, /)

Difference Call: DiffCall (g, /) (only for / ^' ε {9, 15, 17, 19, 20.5, 24, 25, 27})

Average Difference: AvgDiff (g, /)

Positive Fraction, Absolute Call, Difference Call and Average Difference are parameters used by Affymetrix, the definition of which results from the descriptions of the algorithms used in the analysis software by Affymetrix.

2. Exclusion and inclusion rules

2.1 Exclusion of unreliable / uninteresting observations

The measurement of the expression of the gene g at time i is considered unreliable or uninteresting if

• AbsCall (gf, /) = A or

• DiffCall (sf, /) = NC.

First exclusion rule:

All genes g for which all times i one of the above conditions are met are excluded. Second exclusion rule:

All genes g, with DiffCallfg, i) = NC for all i e {9, 15, 17, 19, 20.5, 24, 25, 27} are excluded.

For the remaining genes, all average difference values below 100 are set to 100.

2.2 Selection based on variation of expression

Be

M {g) = max AvgDiff (g, /)

/ e {9,15,17,19,20.5,24,25,27,31}

and

m (g) = min AvgDiff (g, /). i e {9,15,17,19,20.5,24,25,27,31}

Let Q (g) = M (g) lm (g).

Einschlußreqel:

Only those genes g are considered for which either

• M (g) ≥ 500 and Q (g) ≥ 5 or

• M {g) ≥ 600 and Q {g) ≥ 4 or

• M (g) ≥ 700 and Q {g) ≥ 3.5.

Good. 3. Correspondence analysis

The average difference values can be understood as frequencies. Correspondence analysis ([2]) suggests this as a means of examining the interdependencies between times and genes. Figure 2 in the Appendix shows the arrangement of the times; Figure 3 shows the arrangement of the genes. The first two coordinates explain over 82% of the variation in the data. This is highly significant. Test data generated by random and independent genetic permutation resulted in an average value of 29% and a maximum value of 32% over 500 runs.

4. Selected gene environments

In order to compare the expression profiles of different genes, it makes sense to standardize them appropriately. One can imagine the average difference values written in a blackboard (matrix), in which the genes correspond to the rows and the times correspond to the columns. The interpretation of the expression values as frequencies suggests (see [2]) to standardize the rows to sum 1. Be

rs (gf) = ∑ AvgDiff (sf, /)

and

cs (/) = ∑ AvgDiff (g, /). 9

If Average Difference values are arranged in a matrix as above, the rs {g) values correspond to the row totals and the cs (/) values correspond to the column totals. The above genetic standardization of the expression profiles over the 9 points in time results in:

R (g, i) = AvgDiff (gf, /) / rs (gf). This size indicates the relative or percentage expression of the gene g at the time / in relation to the total expression over all times. So it applies

Σ Fl (9, i) = 1 ■

In order to be able to compare two gene profiles R (g, i) and Rief, /), a suitable distance between them is required. One such is the χ ² distance

d (g, et) = ∑ (R (g, i) - R (cf, i) f I Cs (ι) i

where Cs (i) = cs (ι) I ∑, cs (i). In order to eliminate the influence of individual points in time, one can proceed to the jackknife variant of this distance ([3]). This arises from the fact that the χ ² distance is calculated by omitting one of the 9 points in time and then determining the maximum of the values obtained. Be

rs_ / Sy = Σ AvgDiff (g, /) i ≠ j

and for i ≠ j

R-j {g, i) = AvgDiff (gf, /) / rs_ / g).

Then applies

ccts, g = max d_j {g, g ¹ ) j

in which d_ {g, 0) = ∑ (R_j {g, i) - f? _ / \ i) I Cs (i). i ≠ j

With this distance, the next 20 genes became each

Ha5 Keratin AF 020 790

• CDC 20 AB 045 313

• Topoisomerase (DNA) II U 019 5

• CD 53 X 97227

certainly. The results can be found in Tables A to D in the Appendix.

The P values are simultaneous P values calculated from permutation statistics (100 runs) according to [4] (see also [1])). They relate to the null hypothesis that the gene profiles of central genes and the remaining genes are uncorrelated. The distance between AF 020 790 and AB 045 313 is 1.14 with a P value of 0.63. The two genes show clearly different behavior and are not correlated. The distance between AB 045 313 and U 01915 is 0.19 with a P value of 0.01. The expression profiles of the two genes thus show a clear correlation.

The standardized profiles of the 10 genes closest to the center of the corresponding groups can be found in FIGS. 4 to 7. FIG. 8 shows the superposition of the profiles of the AF 020 790 group and the AB 045 313 group. Figure 9 shows the superposition of all four groups. Figure 10 shows the four groups in the plot of the coordinates of the correspondence analysis.

You can clearly see the phase shift. FIG. 11 shows the superimposition of FIG. 2 and FIG. 10. One can see the approximate assignment of the gene groups at the times at which their expression reaches its maximum; the AF 020 790 group is in the direction of dP9 / dP15, the AB 045 313 group and the U 01915 group in the direction of dP27 / dP31 and the X 97227 group in the direction of dP24. Literature for Example 2

[1] Dudoit, S., Fridlyand, J. and Speed, T. P .: Comparison of dicrimination methods for the classification of tumors using gene expression data. Department of Statistics, University of California, Technical report # 576 (2000).

[2] Greenacre, M .: Correspondence analysis in medical research. Statistical Methods in Medical Research 1, 97-117 (1992).

[3] Heyer, L.J., Kruglyak, S. and Yooseph, S .: Exploring expression data: identification and analysis of coexpressed genes. Genome research 9, 1106-1115 (1999).

[4] Westfall, PH and Young, SS: Resampling-based multiple testing: examples and methods for p-value adjustment. Wiley, (1993).

Table A: 20 next genes, i.e. of the genes with a similar expression course to AF 020 790

Dist Pvalues

TC29428_at 0.00000 0 AF 020 790

TC30751_f_at 0.02478 0 AF 021 836

TC16554_at 0.04347 0 AI 642 522

TC27367_s_at 0.04836 0 X 59 252

TC28346 f at 0.09910 0 M 277 34

TC30749_f_at 0.10118 0 M 920 88

TC31893 at 0.10306 0 M 81 445

TC21963_s_at 0.10969 0 U 08215

TC20645_at 0.11307 0 AA 144 272

TC30750 at 0.11928 0 M 92 088

TC32900_at 0.12157 0 AI 642 522

TC21983 at 0.13572 0 AI 180 528

TC23080 f at 0.14349 0 M 31 690

TC31847 at 0.14400 0 AA 791 965

TC28163_f_at 0.14578 0 M 92 088

TC28347_at 0.14599 0 M 277 34

TC30747_at 0.15120 0 X 99 143

TC30748 f at 0.15169 0 M 92 088

TC20781_at 0.16368 0 AI 157 167

TC25414_s_at 0.16374 0 X 14759

TC34643 at 0.16517 0 FROM 033 744

Table B: 20 closest genes to AB 045 313

Dist Pvalues

TC28355_g_ 0.00000 0 FROM 045 313

TC33816_at 0.03130 0 X 62 154

TC27570_g_at 0.03691 0 AC 002 397

TC20077_at 0.04312 0 AK 003 838

TC14788_s_at 0.06107 0 AF 012 710

TC18751_s_at 0.06289 0 X 75 483

TC31432_at 0.06601 0 AI 574 275

TC34455_at 0.06776 0 AW 061 324

TC29394_at 0.07005 0 AF 107 296

TC34036_at 0.07321 0 AF 013 119

TC34235_at 0.08534 0 AW 209 229

TC26388_at 0.08759 0 AV 117 844

TC14392_f_at 0.09082 0 AI 838 080

TC34018 at 0.09420 0 AA 671 429

TC27894 at 0.09489 0 AW 045 665

TC23810_g_at 0.09545 0 AW 049 504

TC18819_i_at 0.10111 0 AI 606 257

TC27344_g_at 0.10685 0 D 86 726

TC34455_g_at 0.11259 0 AW 061 324

TC16956_s_at 0.11376 0 AW227 345

TC34515_s_at 0.11382 0 X 75 983

Table C: 20 closest genes to U 01915

Dist Pvalues

TC15204_s_at 0.00000 0 U 01915

TC23810_g_at 0.03057 0 AW 049 504

TC28934_at 0.03954 0 AA 691 890

TC34455_g_at 0.06010 0 AW061 324

TC17247_at 0.06501 0 AW123 386

TC18751_s_at 0.07500 0 X 75 483

TC27570_g_at 0.09592 0 AC 002 397

TC19890_g_at 0.10362 0 AI 842 612

TC34036_at 0.10647 0 AF 013 119

TC18816_at 0.11960 0 AW 122 331

TC15641_at 0.12080 0 U 93 583

TC34455_at 0.12338 0 AW 061 324

TC34515_s_at 0.12495 0 X 75 483

TC20077_at 0.14660 0 AK 003 838

TC33816_at 0.14974 0 X 62 154

TC34235_at 0.15473 0 AW 209 229

TC19381_at 0.15909 0 AA 152 809

TC26388_at 0.16076 0 AV 117 844

TC20005_at 0.17047 0 D 87 908

TC14788_s_at 0.17370 0 AF 012 710

TC29394 at 0.18604 0 AF 107 296

Table D: 20 closest genes to X 97 227

Dist Pvalues

TC28894_s_at 0.0000 0 X 97 227

TC22483_g_at 0.1049 0 AW 122 039

TC32158_at 0.1178 0 AI 116 109

TC33838_at 0.1303 0 AI 447 446

TC25563_at 0.1344 0 AA 170 203

TC26610_s_at 0.1462 0 D 76 440

TC19112_at 0.1558 0 AF 051 583

TC31080_s_at 0.1583 0 U 188 69

TC28487_at 0.1799 0 D 45 850

TC32132_at 0.1811 0 AA 144 469

TC30937_g_at 0.1840 0 AA 183 595

TC23195_at 0.1851 0 AI 852 199

TC30596_s_at 0.1868 0 M 31312

TC25622_at 0.1878 0 AI 156 274

TC16835_s_at 0.1911 0 M 12 660

TC29751_at 0.1920 0 AA 756 506

TC29692_at 0.1935 0 AI 120 960

TC32774_s_at 0.1942 0 Z 11 974

TC32891_at 0.1951 0 AW 910 263

TC30631_at 0.2025 0 U 56 819

TC28732 s at 0.2031 0 AJ 223 361

Claims

claims

1. A method for diagnosing a hair growth disorder or for determining the likelihood of a hair growth disorder (predisposition) in a mammal, wherein the expression of at least two cyclically expressed genes in a sample is analyzed and wherein (a) one of the genes has the maximum expression level during the anagen , while (b) the second gene has the maximum expression level during catagen or during telogen.

2. A method of diagnosing the effectiveness of treating a hair growth disorder in a mammal by analyzing the expression of at least two cyclically expressed genes in a sample and wherein (a) one of the genes has the maximum level of expression during anagen, while (b) second gene has the maximum expression level during catagen or during telogen.

3. The method according to claim 1 or 2, wherein the expression (c) of at least a third cyclically expressed gene is analyzed and wherein this gene has the maximum expression level during anagen, during catagen or during telogen.

4. The method according to any one of claims 1 to 3, wherein the expression (d) of at least a fourth cyclically expressed gene is analyzed and wherein this gene has the maximum level of expression during anagen, during catagen or during telogen.

5. The method according to any one of claims 1 to 4, wherein the expression (e) of at least a fifth cyclically expressed gene is analyzed and wherein this gene has the maximum expression level during anagen, during catagen or during telogen.

6. The method according to any one of claims 1 to 5, wherein at least one of the genes which have their maximum expression level in the anagen, this has during the middle anagen.

7. The method according to any one of claims 1 to 6, wherein at least one of the genes which have their maximum expression level in the anagen, this has during the early anagen.

8. The method according to any one of claims 1 to 6, wherein at least one of the genes which have their maximum expression level in the anagen, this has the transition from anagen to catagen.

9. The method according to any one of claims 1 to 8, wherein the genes listed under (a), (b), (c), (d) and (e) are representatives of different groups of cyclically expressed genes, the diversity of the groups thereby is characterized in that they have their expression maxima in different time periods and wherein genes belonging to the same group have the same expression profiles.

10. The method of claim 9, wherein the expression of at least one other member from each group of the cyclically expressed genes is analyzed.

11. The method of claim 9 or 10, wherein the expression of at least two other members from each group of the cyclically expressed genes is analyzed.

12. The method according to any one of claims 1 to 11, wherein the sample is a skin sample or consists of epilated hair.

13. The method of claim 12, wherein the skin sample is a scalp sample.

14. The method according to any one of claims 1 to 13, wherein the mammal is a mouse.

15. The method according to any one of claims 1 to 14, wherein the at least one gene (a) is selected from the group of genes which comprise the sequences represented by the Genbank accession numbers listed in Table 1 or the human homologues thereto.

16. The method according to any one of claims 1 to 15, wherein the at least one gene (b) is selected from the group of genes comprising the sequences represented by the Genbank accession numbers listed in Table 4 or the human homologues thereto.

17. The method according to any one of claims 1 to 16, wherein the at least one gene (c) is selected from the group of genes comprising the sequences shown by the Genbank accession numbers listed in Table 2 or the human homologues thereto.

18. The method according to any one of claims 1 to 17, wherein the at least one gene (d) is selected from the group of genes which comprise the sequences represented by the Genbank accession numbers listed in Table 3 or the human homologues thereto.

19. The method according to any one of claims 1 to 18, wherein the at least one gene (s) is selected from the group of genes which comprise the sequences represented by the gene bank deposit numbers listed in Table 5 or the human homologues thereto.

20. The method according to any one of claims 1 to 19, wherein the mammal is a mouse or a human.

21. The method according to claim 20, wherein the at least one gene (a) is the human homologue to the mouse gene mentioned in claim 15.

22. The method according to claim 20, wherein the at least one gene (b) is the human homologue to the mouse gene mentioned in claim 16.

23. The method according to claim 20, wherein the at least one gene (c) is the human homologue to the mouse gene mentioned in claim 17.

24. The method of claim 20, wherein the at least one gene (d) is the human homologue to the mouse gene mentioned in claim 18.

25. The method of claim 20, wherein the at least one gene (s) is the human homologue to the mouse gene mentioned in claim 19.

26. A method for isolating an inhibitor of hair growth disorder, wherein one

(a) analyzing the expression of at least one of the genes listed in the preceding claims under (a) to (e) in a skin sample or in epilated hair which was taken from a mammal with a hair growth disorder;

(b) compares the analysis result from (a) with the expression of the gene in a skin sample or in epilated hair, which is a mammal taken from the same species, this mammal not having this or no hair growth disorder;

(c) the gene whose expression differs by at least a factor of 3 in a mammal with hair growth disturbance compared to a mammal without it or without a hair growth disturbance in step (b), the mRNA transcribed therefrom or the polypeptide encoded thereby or a skin sample, in which the gene is expressed with an abnormal level of expression in contact with one or more test substances under conditions which allow binding of the test substance to the gene, the mRNA or the polypeptide or to other genes, mRNAs or polypeptides in the skin sample; and

(d) isolating the test substance (s) which leads to a reduction in the factor in the expression in step (b).

27. The method of claim 26, wherein the expression of the gene is normalized against the expression of a housekeeping gene.

28. The method according to any one of claims 1 to 25, wherein the hair growth disorder is an alopecia.

29. The method of claim 28, wherein the alopecia is Alopecia androgenetica.

30. The method according to any one of claims 1 to 27, wherein the hair growth disorder is hirsutism.

31. The method according to any one of claims 24 to 28, wherein the test substances or the gene are coupled to a readout system or wherein a readout system is added to the test mixture and wherein the readout system after binding of the test substance (s) to the gene , the mRNA or (poly) peptide provides a detectable signal.

32. The method according to any one of claims 26 to 31, wherein the test substances are low molecular weight substances, peptides, aptamers, antibodies or fragments or derivatives thereof.

33. The method according to any one of claims 26 to 32, which is a high-throughput method.

34. The method of claim 33, which is computer-assisted.

35. The method according to any one of claims 26 to 34 further comprising improving the pharmacological properties of the identified inhibitor, wherein

(e) the binding site of the inhibitor to the gene, the mRNA or the polypeptide and optionally the binding site of the gene, the mRNA or the polypeptide to the inhibitor are identified;

(f) modifying the binding site of the inhibitor and the gene, mRNA or polypeptide by molecular modeling; and

(g) modifies the inhibitor so that its binding specificity or binding affinity or binding avidity for the gene, the mRNA or the polypeptide is increased.

36. The method according to claim 35, wherein the binding sites in step (a) are determined by site-specific mutagenesis.

37. The method according to any one of claims 26 to 34 or according to claim 35 or 36 further comprising improving the pharmacological properties of the identified inhibitor or the improved inhibitor, wherein the (poly) peptide as the lead structure is further modified to

(i) a modified active center, a modified activity spectrum, a modified organ specificity and / or (ii) an improved activity and / or (iii) a reduced toxicity (an improved therapeutic index) and / or (iv) reduced side effects and / or (v) a delayed start of the therapeutic efficacy, the

Length of therapeutic efficacy and / or (vi) changed pharmacokinetic parameters (absorption, distribution,

Metabolism or excretion) and / or (vii) modified physicochemical parameters (solubility, hygroscopic

Properties, color, taste, smell, stability, condition) and / or (viii) improved general specificity, organ / tissue specificity, and / or (ix) optimized administration form and route through

(i) esterification of carboxyl groups or (ii) esterification of hydroxyl groups with carboxylic acids or (iii) esterification of hydroxyl groups to, for example, phosphates,

Pyrophosphates or sulfates or "hemisuccinates" or (iv) formation of pharmaceutically acceptable salts or (v) formation of pharmaceutically acceptable complexes or (vi) synthesis of pharmacologically active polymers or (vii) introduction of hydrophilic groups or (viii) introduction / exchange of Substituents in aromatics or

Side chains, change of the substituent pattern or (ix) modification through the introduction of isosteric or bioisosteric

Groups or (x) synthesis of homologous compounds or (xi) introduction of branched side chains or (xii) conversion of alkyl substituents to cyclic analogues or (xiii) derivatization of hydroxyl groups to ketals or acetals or (xiv) N-acetylation to amides, phenyl carbamates or (xv) synthesis of Mannich bases, imines or (xvi) conversion of ketones or aldehydes into Schiff bases, oximes,

Acetals, ketals, enol esters, oxazolidines, thiozolidines or combinations thereof.

38. The method according to any one of claims 26 to 37, wherein the identified or improved inhibitor is further pharmacologically improved by peptidomimetics.

39. The method according to any one of claims 26 to 38 further comprising the manufacture of a medicament or medical preparation, wherein the inhibitor obtained is formulated with a pharmaceutically acceptable carrier or diluent.

40. The method according to any one of claims 26 to 39, wherein the inhibitor of hair growth disorder is an activator of the transcription or translation of the gene, the mRNA or an activator of the activity of the polypeptide.

41. The method according to any one of claims 26 to 39, wherein the inhibitor of hair growth disorder is an inhibitor of the transcription or translation of the gene, the mRNA or an inhibitor of the activity of the polypeptide.

42. Use of an inhibitor identified by the method according to one of the preceding claims for the manufacture of a medicament or medical device for the prevention or treatment of hair growth disorders.

43. A composition comprising one or more of the inhibitors which have been identified or improved by the method according to any one of claims 24 to 38.

44. The composition of claim 39 which is a pharmaceutical composition.

45. The composition of claim 39 which is a diagnostic composition.