DE102014018663A1 - Detection of viable cells for the review of drug information in drug approval procedures - Google Patents

Abstract

The invention relates to a detection of viable cells in the context of a verification of drug data from drug approval procedures.

Description

The invention generally relates to a detection method for cells capable of reproduction.

The question of whether a given active substance actually shows the alleged effect occurs with the manipulation of drug authorizations observed by government agencies. Adulteration of the ingredients of medicinal products with the addition of ineffective, cheaper substitutes in counterfeit products are common in product piracy. This is in the present application now not the subject of consideration, but the formulation of drugs with possibly patented original active ingredients, which, however, a biochemical effect from the beginning materially missing.

For example, a common drug claim connects aloud Federal Supreme Court Decision "Dipeptidyl Peptidase Inhibitors" X ZB 8/12 dated 11.09.2013 a selection group of drugs, eg. B. all not materially structurally unrelated inhibitors of an enzyme, with a specific medical indication. This indication should indicate, according to popular beliefs, against which disease state the active substance should show an effect in the patient case.

The claim concerning the conspicuous dressing of a drug is reworded in accordance with the revised Federal Constitutional Court decision Paragraph 3 (4) of the PatG: '(Use of an inhibitor (s) of dipeptidyl peptidase (DP IV) enzyme activity to lower blood glucose below the glucose concentration characteristic of hyperglycaemia in the serum of a mammalian sucker organism.'

This technical teaching is considered to be feasible even if only the pharmacist is able to prepare the drug, because the effectiveness itself is not considered to biochemical effects in the cell, but it is patented by the mere indication of indication to which the drug was prepared.

• a) eine Erkrankung wird als Indikation ausgewählt (hier Diabetes),
• b) ein Protein wird als Zielmolekül (Target) erkannt, da dessen biologische Funktion im Krankheitsprozess eine Rolle spielt (hier das Enzym DP IV),
• c) ein Small Organic Molecule (SOM, ein kleines organisches Molekül) wird als Interaktor des Zielmoleküls identifiziert (hier Aminoacyl-Thiazolidide, vgl. die Schrift DE 196 16 486 A1 , Spalte 4, Zeile 31),
• d) das SOM wird augenfällig pharmazeutisch zugerichtet und mit der Indikationsangabe versehen.

The claim according to the BGH decision "dipeptidyl peptidase inhibitors" also gives the current development process of medicinal active substances [cf. Terstappen, GC et al., Nature Reviews Drug Discovery (2007) 6, 891-903 ] again, with the steps:

• a) a disease is selected as an indication (here diabetes),
• b) a protein is recognized as a target molecule, since its biological function plays a role in the disease process (here the enzyme DP IV),
• c) a Small Organic Molecule (SOM, a small organic molecule) is identified as an interactor of the target molecule (here aminoacyl-thiazolidides, see the Scriptures DE 196 16 486 A1 , Column 4, line 31),
• d) the SOM is apparently pharmaceutically prepared and provided with the indication.

The finding according to step b) that a particular molecule plays a role in a particular disease comes, for example, from the correlation of the disease with the change in the expression of the gene coding for the target protein, ie, for. At the level of the messenger RNA of the target protein. Therefore one calls the procedure of the BGH patent claim technically possibly "target based drug screening".

The interactor in step c) may also generally be a biological, e.g. B. be an antibody.

In the context of extensive drug studies on patients worldwide by the administration of drug formulations of different groups of substances that are to have interactor activity, medical observations on patients carried out with regard to the indication of the claim, that is just not with respect to the actual biochemical The effect of the active substance, as has become known, is easily distorted in order to obtain a marketing authorization in certain countries with this false information.

In the present case, it is therefore proposed to verify the medical indications of at least one given active substance asserted on the basis of patient observations without the indication according to patented indication of effect, namely by means of cell detections with at least one active ingredient and with regard to its biochemical action. It can thus be speeded up to verify that the active substance shows any effect in the biochemistry of the cell.

• 1) L-threo-Isoleucyl-Thiazolidid.

In a first step, a library of individual product mixtures is provided, which are characterized in that each individual product mixture contains a certain amount of the at least one active substance to be investigated; to the above BGH decision claim this would be an example according to the Scriptures DE 196 16 486 A1 , Column 4, lines 46 to 47:

• 1) L-threo-isoleucyl-thiazolidide.

It provides the library by at least 1,000 different, pharmacologically meaningful dressings of the at least one Be provided in the form of individual product mixtures as library members.

In a second step, at least one cell capable of replication is brought into contact with a test individual product mixture as a library member containing the at least one active substance to be investigated. The cells may be present in de novo programmed cell or tissue models, in in vitro tissue models, in isolated organs or in in vitro cultured organs, as well as in subsystems of organisms or organisms models.

The cells may also be minimal cells endowed with artificial, in vitro synthesized human minimal chromosomes that are absent in the natural human, and which comprise a non-naturally occurring human gene set optimized for the present verification process For the complete, arbitrary reprogramming of a cell, and thus of an organism with the deletion of the superfluous parts of the old, natural program, is well known to be the essence of synthetic biology [cf. Callaway, Ewen: First synthetic yeast chromosome revealed. Nature News (March 27, 2014) doi: 10.1038 / nature.2014.14941 ; Annaluru, Narayana et al .: Total synthesis of a functional designer eukaryotic chromosomes. Science (4 April 2014) 344 (6179) pages 55-58 ]. Equally, tailor-made synthetic cells may also be those whose total phenotype, as determined by the expression state of corresponding marker gene products, corresponds to that of human embryonic stem cells (cf its patentability ECJ judgment C-34/10 of 18.10.2011 , see also DE 197 56 864 C5 ).

The gene set of the cell is provided by determining responsive genes for the considered, at least one drug. From the viewpoint of the person skilled in the art, in this case a bio-synthesizer, their base sequences must obviously not correspond to the natural base sequences, as long as they are only able to perform the respective functions (input-output transformations) even after reprogramming.

According to the application, it is proposed to use those genes as responsive genes which are suitable for the protein target or the enzyme target according to development step b) of the at least one active substance to be investigated, e.g. As a SOM or a biological can encode, whose expression at the messenger RNA level so according to the above development step b) changes anyway in the onset of the disease.

According to the application, however, responsive genes can also be those genes which code for proteins which complex with the target protein in the cell, or z. B. in signal transduction chains with the target protein exchange information.

On the basis of this patent application, it has been shown on the patent law side that the present target molecule-based development of active substances described above in steps a) to d) is accompanied by target interactor screening by the application of patents containing the respective target Proteins encoding genes have to claim anyway for the protection of the screening method as substances (see, for example, the Scriptures EP 0 939 804 B2 , Claim 1 and the associated protection certificate method with ECJ judgment C-493/12 dated 12.12.2013 to the antibody tabalumab). The influence of the expression of the messenger RNAs of the target proteins themselves by their interactors is obviously not scientifically investigated systematically. For a promising interactor, it is sufficient to interact with the target molecule at the protein level, and to patent this interaction. An approach is increasingly only financially viable if it is to be protected by patenting, and therefore the patentable way becomes the only product development strategy that can still be financed, such as that of the above steps a) to d): This is governed by the above Federal Supreme Court claim displayed.

Thus, according to the application, there is provided: detection of natural or synthetic replicable cells, genetically preprogrammed or de novo programmed to function as an input-output transformation indicative of drug activity, consisting in that upon contact with a library member of a library of a product containing the to be examined, at least one active substance is a cellular reaction in terms of the active ingredient activity.

In this case, the genetic programming of the cells for the function of an active substance activity is the genetic information of the active ingredient-responsive genes
in the cells,
which transforms an input of the cellular contact with a library member into an output of the cellular response in the form of a change in the expression of the at least one responsive gene.

The cellular response of the at least one cell can be recognized by means of a change in gene expression, which can be determined e.g. B. can record with the transcriptome covering microarray, or by changing the cellular architecture. Responsive genes that do with the drug activity and whose expression change can be detected after contact of the cell with a library member prepared in the first step, in the above example of the BGH decision the gene for the enzyme DP IV: gene DPP4 [cf. Arndt, Marco et al., Biochem. Biophys. Res. Commun. (2000) 274 (2): 410-4 ].

In the context of providing a universal empty cytoplasm (I: hardware) for receiving completely re-programmed genetic information, such. B. a completely newly synthesized genome (II: software), a tissue model can also be used a cell assembly with completely new-programmed cells, the genetic de novo software indicating an active substance activity input-output transformation (III: genome function ). The genetic software to be designed de novo can be adapted individually to the respective customer target needs with regard to the genetic organism crisis situation subsystem to be used for the reaction test, without a new organ or tissue system over and over again, while the empty cytoplasm remains unchanged Cell cultures must be created from isolated cells. Thus, it is further provided: a proof whereby the cells are considered individualized or personalized.

The genetic information provided to an empty cytoplasm may be taken to predict the response of a whole organism to a library member from the genome of that whole organism itself without having to provide its cells. Here, an individual-typical, personalized genetic reaction of a replicative test cell is detected on a product mixture.

The cells may be taken to predict the response of a whole organism model to a library member from this whole organism model itself. Thus, it is provided: A proof, wherein the cells are programmed de novo genetically typical of individuals, with the exception that the nucleic acids are materially isolated from the individuals and cloned.

QUOTES INCLUDE IN THE DESCRIPTION

This list of the documents listed by the applicant has been generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

Cited patent literature

• DE 19616486 A1 [0006, 0011]
• DE 19756864 C5 [0014]
• EP 0939804 B2 [0018]

Cited non-patent literature

• Federal Supreme Court Decision "Dipeptidyl Peptidase Inhibitors" X ZB 8/12 dated 11.09.2013 [0003]
• Paragraph 3 (4) of the PatG: "(Use of an inhibitor (s) of dipeptidyl peptidase (DP IV) enzyme activity to lower blood sugar below the glucose concentration characteristic of hyperglycaemia in the serum of a mammalian sucker organism." [0004]
• Terstappen, GC et al., Nature Reviews Drug Discovery (2007) 6, 891-903 [0006]
• Callaway, Ewen: First synthetic yeast chromosome revealed. Nature News (March 27, 2014) doi: 10.1038 / nature.2014.14941 [0014]
• Annaluru, Narayana et al .: Total synthesis of a functional designer eukaryotic chromosomes. Science (4 April 2014) 344 (6179) pages 55-58 [0014]
• ECJ Judgment C-34/10 of 18.10.2011 [0014]
• ECJ judgment C-493/12 of 12.12.2013 [0018]
• Arndt, Marco et al., Biochem. Biophys. Res. Commun. (2000) 274 (2): 410-4 [0021]

Claims (5)

Detection of viable cells that show a cellular response on contact with a mixture of products, with the steps of: a) providing at least one drug substance, b) identifying the protein target molecule or enzyme target molecule, as its protein-level interactor, this drug drug has been structurally engineered, c) identifying at least one gene that is responsive to said drug in the cell, preferably the gene coding for the protein target molecule or the enzyme target molecule, d) providing natural or synthetic cells capable of reproduction, genetically preprogrammed or de novo programmed to function an active substance activity indicating input-output transformation, consisting in that when in contact with a single product mixture of the at least one active substance to be tested a cellular response occurs in terms of drug activity, in which genetic programming for the function of an active substance activity the genetic information of at least one responsive gene for the at least one active ingredient in the cells, providing an input of the cellular contact with a single product mix as a member of a mix product library into an output of the cellular reaction transformed in the form of a change in the expression of the at least one responsive gene, and e) Detection of the active substance activity indicating input-output transformation by detection of the reacting cells. Detection according to claim 1, wherein the cells are present in de novo programmed cell or tissue models, in in vitro tissue models, in isolated organs or in in vitro cultured organs, as well as in subsystems of organisms or organisms models. Detection according to one of the preceding claims, wherein the cells are programmed de novo genetically typu individual typical, so it is exempted that nucleic acids are materially isolated from individuals and cloned. Detection according to one of the preceding claims, wherein the cells are minimal cells equipped with artificial in vitro synthesized human minimal chromosomes, which do not occur in natural humans, and which are optimized for the present detection method, not occurring naturally in its entirety comprising at least that for the at least one active ingredient at least one responsive gene. Detection according to one of the preceding claims, wherein the at least one active ingredient is L-threo-isoleucyl-thiazolidide and the at least one responsive gene is the gene DPP4, coding for the target protein of this at least one active ingredient, namely the enzyme dipeptidyl peptidase (DP IV.