Abstract:
The invention relates to a method for producing an extract containing at least 40% (m/m) of polyphenols from  Cistus creticus  L., said method comprising the following steps: plant parts are extracted from  Cistus creticus  L. using an extraction agent selected from the group comprising water, alcohols and mixtures thereof; the extraction residues are removed; the extraction agent is at least partially removed; redissolution is carried out in an aqueous solvent and the undissolved constituents are removed; and selective enriching is carried out by means of either a) a two-phase extraction, and b) a membrane filtration.

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
     This application is a national stage filing of PCT Application No. PCT/EP2009/055487 filed May 6, 2009, which claims priority to European Patent Application No. 08155699.5 filed May 6, 2008, each of which is hereby incorporated herein by reference. 
     FIELD OF THE INVENTION 
     The present invention refers to enriched extracts from  Cistus , compounded formulations thereof, processes concerning their manufacturing, and their usage. 
     BACKGROUND OF THE INVENTION 
     Within the family of Cistaceae,  Cistus  forms a genus of 18 to 20 species. The evergreen subgenus  Cistus creticus  L. ssp.  eriocephalus  reaches a height of up to 1 m and smells aromatic. Its egg-shaped lanceolate leaves show an alternate pattern, while its flowers have five petals and a radial symmetry, which is characteristic for the whole family. The light pink, pink, or light purple colors are typical for the subtype. Preferred habitat is the eastern Mediterranean region. 
     Folk medicinal usages have been versatile and ranged from being used as adjuvant for topical, allergy-related itching, as well as for preventing and managing wounds caused by bacterial infections and mycosis. Traditionally, aqueous decoctions from the aerial plant parts from spring have been used for this. Similarly have recent publications reported about the successful usage of  Cistus  solutions for the treatment of neurodermititis (AD), acne vulgaris, as well as for treating inflammatory diseases of the mouth and throat.  Cistus  is also well-known for its usage as tea (e.g. Cystus® Bio Teekraut). Recommended dose here is 2.5 g of  Cistus incanus  for 1 liter of hot water. According to our analysis, up to 1 g of dissolved extractives with ca. 16% polyphenols can thus be ingested. Other providers of  Cistus , who give no detailed information on the species, obtain their raw material from wild collectings, which in consequence lead to unsteady results in both, extractives (0.7-1.5 g), as well as polyphenol contents (11-21%). Such, not-standardized mixtures represent the natural polymorphism of the plant and offer no basis for the manufacturing of a standardized plant extract. 
     A detailed specification of the varieties and/or subspecies has only recently become urgently necessary and relevant because of a legal distinction in the plant varieties&#39; rights (PVR) between  Cistus incanus L . ssp. Pandalis®, or the brand name Cystus®, and  Cistus creticus  L. ssp.  eriocephalus  (also known as  Cistus incanus  L. ssp.  tauricus ). It is very difficult to distinguish between the different types, especially for persons lacking expert knowledge. 
     SUMMARY OF THE INVENTION 
     The oldest known preparations of  Cistus  deal with labdanum, a resin, which was mostly gained from the leaves and stems of  Cistus ladaniferus L., Cistus monspliensis  L., and  Cistus incanus  L. Because of its pleasant aroma, it was often incensed. Other utilizations included that of an expectorant to treat catarrhs of the respiratory tract, as well as plastri and unguenta (medical dressings) for wound management purposes. 
     Phytochemical analyses of genus  Cistus  have therefore traditionally concentrated on analyzing its lipophilic substances, especially of its essential oils and resins. Here, we find the normal structures of the terpene metabolism, such as mono-, sesqui-, and diterpenes, as well as alcohols and esters. Other analyses have focused on the polyphenol fractions of its flavonoids and tanning agents. Kaempferol, quercetin and apigenin proved to be the most basic parts of the structures. Samples of  Cistus creticus  ssp have proven to be outstandingly rich in polyphenols. Surprisingly, our analyses have revealed that the polyphenol fingerprint of  Cistus creticus  ssp.  eriocephalus  resembles another, well-known pattern: Similar to extracts of green tea, in the area of the important oligomer fractions, derivates of catechin, such as gallocatechin or even epigallocatechingallat (EGCG) can be found. The total content of polyphenols in the plant is generally rated at 4%, while the concentration in the leaves is substantially higher. 
     Polyphenols are widely known for their antioxidant behavior, which refers to their potential to combat free radicals. In the human body, these are especially “reactive oxygen species”, which are often caused by environmental influences (UV-rays, chemical noxes), but also by an unbalanced, unhealthy diet. According to latest state-of-the-art knowledge, many diseases can be linked to “reactive oxygen species”. Cell damages attributed to radicals can lead to a number of diseases (e.g. immune diseases, mental exhaustion, arthritis) if they are not limited by “antioxidant scavengers”. Thus, measuring the antioxidant capacity oftentimes delivers decisive values. High contents in polyphenols simultaneously indicate a high antioxidant potential. 
     Current research on an extract of the pink rock rose (CYSTUS® 052; manufactured from  Cistus incanus  PANDALIS, 26% of polyphenols, less than 2% of monomers (gallic acid, epigallocatechin, catechin and epicatechin)) shows its ability to significantly inhibit the increase of influenza viruses in in-vitro experiments, without producing any occurrence of resistance (Ehrhardt C. et al., Antiviral Res., 76, 38-47, 2007). Likewise, these results could be verified in an in-vivo (mouse model) experiment (Droebner, K. et al., Antiviral Res. 76, 1-10, 2007). 
     Commercial products such as  Cistus incanus  capsules (Co. LR Health and Beauty Systems) generated from simple aqueous extracts (20-25% polyphenols; drug-extract-ratio of 2-4:1), represent the current technical state-of-the-art. Own analyses have confirmed that aqueous extractions from  Cistus incanus  ssp.  tauricus  or  Cistus creticus  L. ssp.  eriocephalus  can produce extracts with 20-28% polyphenol contents, depending on the quality of the used raw materials. 
     In order to be able to evaluate and compare different formulations of  Cistus , a clear definition of the species is necessary. Furthermore are both, specifications concerning the manufacturing (extractant; drug-to-extract ratio), as well as an analytical characterization (polyphenols), necessary requirements for a standardized product. Only a reproducible, standardized extract quality will ensure a stable therapeutic scheme for therapeutical usages. The invention was aimed at providing a method for manufacturing a  Cistus  extract with a high content, which will—in a reproducible way—produce standardized extracts of preferably a minimum content of 40% (w/w) polyphenols. 
     This task is accomplished by a method of manufacturing an extract from  Cistus  using the following steps:
         Extracting of plant parts of  Cistus L . with solvents selected from the group of water, alcohols and mixtures thereof   removal of extraction residues   at least partial removal of extractants   Redissolving into an aqueous solvent and removal of indissoluble compounds   selective enrichment by means of at least one of the following steps:
           a) a two-phase extraction   b) a membrane filtration   
               

     Preferable for such a manufacturing procedure will be a constant quality of plant raw material. This can be achieved by analyzing its contents in polyphenols (stated with the help of Folin&#39;s reagent and calculated as gallus acid according to Method of European Pharmacopoeia, chapter PH. EUR. (2.8.14)), and afterwards by mixing different batches. 
     In order to obtain a higher proportion of leaf material over stalk fractions, the drug is ground as well as air-swept. The leaves have a higher content of polyphenols. Thus, an improvement of 30% can be reached.  Cistus creticus  L. ssp.  eriocephalus  is preferred. Preferably, aerial plant parts are used. 
     Generally, raw materials with a content of a minimum of 12% polyphenols are preferred. 
     Water, methanol, ethanol, 1-propanol, 2-propanol, and mixtures thereof have proven suitable as extractants. Preferably, the content of alcohol does not exceed 50% (v/v), preferably not more than 40% (v/v), or not more than 30% (v/v). 
     One possible embodiment of extraction uses an elevated level of temperature. Temperatures ranging from 40° to 80° C. are especially preferred in order to gain a high yield of polyphenols. 
     After extraction, drug residues are removed, which can be done either by filtration or by suction and later squeezing of the drug residue. Further possible methods are known by experts. 
     Afterwards, the extractant is at least partially removed from the obtained extract. This can be achieved by a rotary evaporator which removes the solvent, or with the help of a panel evaporator. A gentle treatment is preferred. 
     After removal, the ratio of the dry substance is preferably more than 50% (w/w). 
     In a next step, the remaining residues are redissolved. Here, especially water or mixtures from water and alcohol are suitable. Preferably, the redissolver consists of a minimum of 50% of water. 
     After redissolving, the remaining residue can be removed by methods such as filtration, suction, decanting, or the likes, and then be disposed. The remains will contain at least a part of the tannins (little bioactive polyphenols). 
     Afterwards, a step of enrichment follows. Either a two-phase extraction or a membrane filtration can be used. 
     The two-phase extraction can be a liquid-liquid extraction. n-Butanol has proven especially useful as an extractant for this liquid-liquid extraction. 
     Alternatively, or in addition to that, it is possible to conduct a solid-phase extraction. An extraction using adsorption resins is especially useful for this. Typical adsorption resins are for example non-ionic hydrophobic divinylbenzen-copolymers, aliphatic ester polymers and formophenol polymers. Such adsorbers are commercially traded by the name of Amberlite®. Adequate products are types XAD2, XAD4, XAD7HP, XAD16, XAD761, or XAD1180. Also resins produced by other manufacturers such as Diaion (SP-series), or Bayer (Lewatite®), or Miontech (P-series), can be used, as long as they are characterized as being analogous. 
     Resin type Amberlite® XAD7HP is especially preferred. 
     Alternatively, a membrane filtration can be conducted. Pore-defined materials made from polyestersulfon, polypropylene, polytetrafluoroethylene, or cellulose acetate are especially useful as membranes. Appropriate pore-sizes range from 70-100 kDa. The membranes can be installed in several layers into filter panels. Thus, ca. 60% of the polyphenols remain in the retentate (enriched product phase). Membranes made of polyestersulfon are especially preferred. 
     During a liquid-liquid extraction, the polyphenol-containing, aqueous phase is treated with an organic solvent such as n-butanol. In the course of intense mixing, the polyphenol substances transfer into the organic phase, which is then further treated. 
     When conducting a solid-phase extraction, the polyphenols selectively remain on the solid phase, before they can be diluted and separated as a liquid phase via an elution change to a more lipophilic solvent, such as ethanol. The organic phase is then further processed. 
     A membrane filtration separates polyphenols according to their size by pores. Secondary metabolites are held back by pores by means of their size. Together with the filtrate, other attendant substances such as salts and simple acids are separated. An enriched polyphenol fraction remains in the retentate. 
     Afterwards the solvent is removed from the extract by means of a vacuum, which is then dried in order to gain a dry extract. Drying should proceed as gently as possible. 
     Body of invention is also the extract obtained by the aforementioned procedure. 
     In its preferred embodiment, the extract is characterized by at least one of the following features:
         a native drug-to-extract ratio (DEV native) of 4-40:1   an ORAC value of bigger than 3000 μmol trolox equivalent/g   a total polyphenol content of more than 40%, preferably &gt;60% (w/w) and/or   a content of monomers of at least 2% (w/w)       

     Surprisingly, it was found that the invented  Cistus  extract exerts its effect via an inhibition of NF-κB. 
     NF-κB is an important specific transcription factor, has many target genes, and exerts many different effects. The activation of NF-κB is held for playing a critical role in the development of inflammations. Because of its many functions, NF-κB is linked to a large number of diseases. It is often unclear in how far the activation of NF-κB actually causally affects the progression of a disease. Since such a role is more and more seen as probable, the parts of NF-κB signaling pathways have in the meantime become important target structures for the development of new drugs. NF-κB can be attached to a specific DNA-motif, the so-called κB-motif, which consists of about ten base pairs. In the vast majority of cases, attaching NF-κB to the DNA-motif will lead to an increased transcription of the depending genes. At the moment it is thought that about 200 different genes are regulated by NF-κB. Among these are many cytokines (e.g. TNF-κ, and IL-1β), as well as adhesion molecules, which play an important role in regulating the immune system, especially with regard to inflammatory reactions. 
     Amongst others viral anti genes (e.g. lipopolysaccarides) belong to the stimuli which are able to trigger an activation of NF-κB. 
     Allergic inflammatory reactions are also set off via this NF-κB signal cascade, and can thus be reduced with an inhibiting influence. 
     So-called A549 lung cells were used as test models. The effects of the invented  Cistus  extract on the transcriptional activity of NF-κB were examined. The cell system was connected to a luciferase measuring system. After having measured the blank values, the synthetic substance PMA was added as stimulus which enhances the luciferase gene expression in A549 cells significantly.  Cistus  extracts in varied doses were introduced into the system. A clear dose-dependent inhibition could be shown. 
                                                                                             Concentration of  Cistus  extract [μg/ml]                1   10   50   100   150                        Inhibition of NF-κB [%]   none   none   22   42   65                    
Since A549 cells possess binding sites for two possible transcription factors —either for NF-κB or for AP-1—also the AP-1 binding site was examined with regard to its specifity. After having added a AP-1 luc plasmid to measure the activity of luciferase, the following results were obtained:
 
                                                                                             Concentration of  Cistus  extract [μg/ml]                1   10   50   100   150                        Inhibition of AP-1 [%]   none   none   +52   +50   +92                           (activation)                    
An activation of the AP-1 transcription factor could be stated. The results prove that it is a specific NF-κB induced inhibition, and that errors in measuring can be ruled out. The negative cytotoxicity test also demonstrated the proven results.
 
     In order to verify the newly discovered mode of action via NF-κB, another model was chosen, where NF-κB itself has an effect on TNF-alpha (tumor necrosis factor-alpha), which in turn is known from anti-inflammatory research. Afterwards it could be tested whether  Cistus  extract influences a NF-κB induced activation of TNF-alpha (canonic pathway). A cloned cell line 5.1 was used, in which there was one luciferase gene from promoter HIV-1-LTR. The cells were pre-incubated with increasing extract doses, and stimulated with TNF cells for 6 hours. Afterwards, the cells were separated and the report activity in the luciferase model was measured. A dose-related inhibition of the HIV-1-LTR-gene transcription, which depends on TNF-alpha, could be shown (see  FIG. 1 ). Here, the invented extract, with &gt;40% polyphenols, turned out to be significantly more potent than an aqueous extract with &gt;20% polyphenols, which represents the current technical state-of-the-art. 
     The influence of the invented extract on the NF-kappaB signaling pathway was verifiably confirmed. 
     The method produces an extract better suitable than former extracts for the following usages:
         Treatment or prophylaxis of viral diseases   Treatment or prophylaxis of allergy-related inflammatory reactions, especially of allergic rhinitis   Treatment or prophylaxis of disease patterns that accompany a common cold, such as rhinitis, sinusitis, laryngitis, chorditis   Treatment of postviral fatigue syndrome   Treatment of influenza, cholera, clostridic myositis, enteritis necroticans, herpes infections.       

     Subject of the present invention is also an extract formulation containing the invented extract. 
     Subject of the present invention is also a pharmaceutical formulation or a food/dietary supplement, containing the invented extract OR the invented extract formulation. 
     The first contact between a virus and a potential host usually takes place on the mucosa. Accordingly, a local application of the extract onto the mucosa leads to good results. In addition to common treatments in the mouth and throat as a lozenge or as a mouthwash against tonsillitis or gingivitis, treatment surprisingly also showed to be effective against disease patterns that frequently accompany a cold, such as inflammations of the nasal mucosa, the sinuses, the larynxes, and the vocal chords. 
     Surprisingly, it was also found that the invented  Cistus  extract has a high inhibitory effect on neuraminidase activity. 
     Neuraminidases are a family of enzymes of influenza viruses (type A and B) and other viruses that cut terminal sialic acidic rests from glycoproteins on cell surfaces of virus host cells, or viruses themselves. This procedure promotes the detachment of daughter viruses from infected cells. So-called neuraminidase-inhibitors are drugs that decrease the procedure of detaching from the host cell after a viral infection, and thus reduce the (systemic) free viral load. It is generally assumed to be important to take the drug immediately after the first symptoms, since the body&#39;s own immune defense at this point is not exhausted yet, but can be boosted by a medicinal reduction of the virus load. Both, Tamiflu® (Oseltamivir) and Relenza® (Zanamivir), which are hoped to help control a potential pandemic of bird flu (virus H5N1), belong to this type of drugs. Unfortunately, such chemical neuraminidase-inhibitors often cause side effects such as nausea, stomach pain, and vomiting. Furthermore have first resistances against these drugs been observed. 
     Since plant extracts possess a well-known benefit-to-risk profile even over longer periods of intake, the invented  Cistus  extract was tested in comparison to Zanamivir in an in-vitro enzyme model (NA-Star®) according to Wetherall N T, Trivedi T, Zeller J, Hodges-Savola C, McKimm-Breschkin J L, Zambon M, Hayden F G. Evaluation of neuraminidase enzyme assays using different substrates to measure susceptibility of influenza virus clinical isolates to neuraminidase inhibitors: report of the neuraminidase inhibitor susceptibility network. Clin Microbiol. 2003 Feb; 41 (2): 742-50. 
     In order to show the effectiveness of the invented extract, the neuraminidases of the following species were tested: 
                                         IC 50 [mg/mL]  Cistus                 Extract   IC 50 [μM]       Virus Species   60% Polyphenols   Zanamivir                     Clostridium perfringens     36.2   &gt;&gt;200         Vibrio cholerae     43.1       200       Influenza A H1N1   36.7      &lt;&lt;0.01                    
For the invented extract, a remarkable neuraminidase inhibition could be demonstrated, independently from the tested virus species. Thus, the following application areas arise: influenza, cholera, clostridium-myotis, enteritis necroticans.
 
     In addition, a quite universal usability of the invented extract on viral diseases could be shown with herpes simplex viruses type 1 (HSV-1): 
     In order to define the virucidal activity, a viral suspension is exposed to the extract and then the infectiousness of viruses in the cells is measured. Ethanol is used for positive controls. 
     The definition of virustatic activity is then identified via the supernatant of infected human cells after inoculation with extract. Aciclovir is the accepted positive control. 
                                                           Virus Species   IC 50 [μg/mL]               Herpes Simplex     Cistus  extract   IC 50 [μg/mL]   IC 50 [μg/mL]       Virus I   60% Polyphenols   Aciclovir   Ethanol                                Virucidal   3.5   ineffective   80,000       Virustatic   10.3   0.2   ineffective                    
For the invented  Cistus  extract with 60% of polyphenols both, a remarkable virucidal as well as virustatic activity could be demonstrated.
 
     Surprisingly, in practical application even an effect on the postviral fatigue syndrome could be shown. The administration of  Cistus  drops (20 drops three times a day), manufactured according to example 10, showed a significantly faster recovery after the diagnostic finding of an influenza infection within 2-3 days than without medication (4-7 days) in three male test persons aged 40-42 years old. 
     There are many names for the postviral fatigue syndrome, such as royal-free disease, myalgic encephalomyelitis, epidemic neuromyastheny, chronic mononucleosis, chronic Epstein-Barr virus, chronic exhaustion syndrome, or, to keep it short, postviral syndrome. Current research suggests that the disease could be caused by special viruses, such as the Epstein Barr-virus, also initiator of infectious mononucleosis. The symptoms oftentimes only show after an acute infection (often with fever, shivering, bodily pain, enlarged lymph glands, and exhaustion) has already decreased. After the disease has seemingly ended, the patient begins to feel uneasy once again and different ailments will from then on persistently remain. Main symptoms are profound weakness, pain of the muscles, problems with memory and concentration, exhaustion and a persisting or frequent flu-like feeling. 
     Further distinguishing between the numerous types of postviral fatigue syndromes is not too relevant at this point. The disease is diagnosed and treated according to normal principles. Treatment can possibly take several months with slow progressions of the disease, though in many cases an autonomous improvement of the symptoms can be seen. 
     Further application areas are influenza (also H5N1 and H1N1), and herpes simplex infections. 
     The invented  Cistus  extract formulations have inhibiting influence on the NF-κB signaling transduction and can thus be used against (local) allergic infectious reactions in the mouth or throat. Because of its extra activity as neuraminidase inhibitor, appropriate extract formulations can also be used for the treatment and prophylaxis of rhinitis and sinusitis, as well as against postviral fatigue syndromes. 
     Tinctures, syrups, brushings, mouthwashes, (nasal) drops, nasal sprays, powder inhalations, or inhalation solvents are all suitable forms for applying the invented extract. At the same time, dried forms of the invented extract are obtainable for intake as capsules, tablets, dragées, pastilles, soft tablets or lozenges, but also redissolved as a powder, granulate or as an effervescent formulation. In order to have a high concentration locally quickly available, a melting tablet or lyophilizate is especially useful. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  shows effects of  Cistus  extracts on TNF-alpha induced NF-kappaB transcription. 
         FIG. 2  shows an HPLC-fingerprint of the extract according to example 5. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The underlying HPLC-requirements are as followed: 
     Binary high pressure gradient system, i.e. consisting of: 
     
       
         
               
               
             
           
               
                   
               
             
             
               
                 2 pumps: 
                 Model 510 or 515 (Co. Waters) 
               
               
                 Injector: 
                 Autosampler W 717 (Co. Waters) 
               
               
                 Column: 
                 Steel column 250 × 4.6 mm Luna Phenyl-Hexyl, 5 
               
               
                   
                 μm (Co. Phenomenex) 
               
               
                 Column oven: 
                 Jetstream Plus (Co. VDS Optilab) 
               
               
                 Detector: 
                 Tunable Absorbance detector W 2487 (Co. Waters) 
               
               
                 Integrator: 
                 HPLC evaluation software 
               
               
                 Fluxing agent: 
                 A: 180 ml acetonitrile, 40 ml acetic acid and 4.0 
               
               
                   
                 ml of EDTA-solvent are filled into a 2000-ml- 
               
               
                   
                 volumetric flask and filled up with water R almost 
               
               
                   
                 completely. After raising the temperature to room 
               
               
                   
                 temperature, water R is added up to the 
               
               
                   
                 calibrations mark. 
               
               
                   
                 B: 800 ml acetonitrile, 20 ml acetic acid and 2.0 
               
               
                   
                 ml EDTA-solvent are filled into a 1000 ml 
               
               
                   
                 volumetric flask and filled up with water R almost 
               
               
                   
                 completely. After raising the temperature to room 
               
               
                   
                 temperature, water R is added up to the 
               
               
                   
                 calibrations mark. 
               
               
                 Flow: 
                 1.0 ml/min, gradient 
               
               
                 Lingering volume: 
                 2.45 ml (mode routine 2) 
               
               
                   
               
             
          
         
       
     
     
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
               
               
                 Flow rate 
                   
                   
                   
                   
               
               
                 [ml/min] 
                 Time [min] 
                 Eluent A [%] 
                 Eluent B [%] 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 1.0 
                 0 
                 100 
                 0 
                 Separation 
               
               
                 1.0 
                 10 
                 100 
                 0 
               
               
                 1.0 
                 25 
                 68 
                 32 
               
               
                 1.0 
                 35 
                 68 
                 32 
               
               
                 1.0 
                 40 
                 100 
                 0 
                 Conditioning 
               
               
                 1.0 
                 60 
                 100 
                 0 
               
               
                   
               
             
          
         
       
     
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Column temperature: 
                 35° C. 
               
               
                   
                 Injection volume: 
                 10 μl 
               
               
                   
                 Detection: 
                 Absorption in UV-light at 278 nm 
               
               
                   
                   
               
             
          
         
       
     
     The invention will be illustrated further with the help of the following examples: 
     EXAMPLE 1 
     Evaluation of the  Cistus  Raw Material 
     Regrowing, aerial plant parts (leaves, flowers, stems) are used as raw material for the drug. They are dried gently, shredded to an extraction size of about 10 mm, and separated from their stalk fractions by air sweeping. 
     1000 g primary plant extract together with hot water (90°) are put into a percolator and extracted for five hours. After extraction time, it is filtered off, and the extraction is repeated. For the second run, the extraction time takes 3 hours. Afterwards, both extracts are combined and concentrated to a spissum in a rotary evaporator. Now the spissum extracts of each  Cistus  species is analyzed via the DPPH-method with regard to their antioxidant potential. 
     This method is based on a redox reaction with the stable radical 2.2 diphenyl-1-pikrylhydracyl (DPPH). Thus, the extract&#39;s ability to act as a direct antioxidant can be measured. The DPPH radical has a purple color, which is due to the unpaired electron at the nitrogen atom. As soon as the radical attaches itself to one hydrogen atom of a scavenger, the reduced DPPH-H (2.2 diphenyl-1-picrylhydrazin) develops. The antioxidant effect can then be measured by a photometrical measurement of the decrease of the absorption at 517 nm. An inhibition rate of 50% is given. 
                                               Cistus  species   IC 50 [μg/mL]                             Cistus ladanifer  L.   7.9 ± 0.7             Cistus florentinus     3.1 ± 0.2             Cistus monspeliensis  L.   2.8 ± 0.2             Cistus canescens     2.0 ± 0.2             Cistus creticus  L. ssp.  creticus     1.3 ± 0.2             Cistus creticus  L. ssp.  eriocephalus     1.0 ± 0.1                        
Result: aqueous extracts from the species  Cistus creticus  L. are superior to the other species when looking at their antioxidant potential. Especially potent are extracts of the subspecies  Cistus creticus  L. ssp.  eriocephalus.  
 
     EXAMPLE 2 
     Aqueous Raw Extract 
     An aqueous extract is manufactured from 10 kg of the drug Herba  Cistus creticus  L. ssp.  eriocephalus  (polyphenol content of 12.3%), and 20 times as much water at 80° C. by exhaustive percolation. After a vacuum-concentration, the aqueous extract has a dry matter ratio of 60%. The yield of native extract amounts to 5 kg. The polyphenol content amounts to 20.0% calculated to the native extract. 
     EXAMPLE 3 
     Ethanolic-Aqueous Raw Extract 
     15.5 kg of the drug Herba  Cistus creticus  L. ssp.  eriocephalus  (polyphenol content of 14.6%) are exhaustively extracted two times in the percolator at 40° C. with ethanol 40% (v/v) at 1:8. After having separated the eluates from the drug, they are combined and filtered before being gently freed from the solvent at 50° C. in a vacuum. 6 kg aqueous spissum extract with a dry matter ratio of 65% (=3.9 kg native extract) result from this procedure. The polyphenol content amounts to 31.4% in relation to the native extract. 
     EXAMPLE 4 
     Extract Re-Solution 
     5.8 kg spissum (3.77 kg native extract) of the extract—as obtained according to example 3—are redissolved to 20% of dry matter and intensely homogenized for 60 minutes, by stirring. Afterwards, the preparation rests for 4 hours at 10-15° C. Insoluble sediment (5% of the total content) has settled on the ground. This sediment (tannin fraction) shows a polyphenol content of 22% in relation to the native extract. 
     The supernatant (product phase) is stripped from above and cleared through a CP1KS filter panel (yield: 95%). The polyphenol content was 32.0% in relation to the native extract. 
     EXAMPLE 5 
     Solid-Phase Extraction 
     The extract solution according to example 4, containing 3.58 kg of native extract as ca. 20% solution, is added to a column, which is filled with 50 L adsorber resin (Amberlite® XAD7HP). The first cycle is separated and then discarded. Then, the column is cleaned with 3 channel volumes (150 L) of water. The elution of the adsorbed secondary  Cistus  metabolites takes place with 100 L of ethanol 96% (v/v). The ethanol-water eluate is collected, filtered, removed from its solvent by vacuum, and then evaporated to a spissum. 2 kg of spissum with a dry matter of 65% (=1.3 kg native extract) result. The polyphenol content amounted to 65.0% in relation to the native extract. The ratio of monomers (theogallin, epigallocatechin, catechin, epicatechin, and also epigallocatechingallat) amounts to 3.6%. The extract is characterized by a specific HPLC fingerprint (see  FIG. 2 ). 
     EXAMPLE 6 
     Dry Extract 
     Spissum extract—as obtained according to example 5—is spray-dried. For this, the spissum is adjusted to a dry matter of 40% with water and loaded to the spray dryer, which has an airstream temperature of 180° C. A fine powder results after the atomization in the tower, which leaves the tower in an airstream at 100° C. However, the product temperature of the extract does not exceed 55° C. A fine powder (95% &lt;0.315 mm) results, with a loss on drying of 4%. The polyphenol content was 64.8% in relation to the native extract. 
     EXAMPLE 7 
     Membrane Filtration 
     100 g of spissum extract—as obtained according to example 3—(65 g native extract ratio) are redissolved with demineralized water to 5% dry matter and cleared from sediments. Afterwards, the pH-value is set to 4.5-5.0. 
     During two hours, the solution is separated into two fractions via a tangential flow filtration using a 100 kDa membrane (polyethersulfone). A filtered permeate phase as well as a retained retentate phase constitutes. 
     Both phases are evaporated to a spissum in a vacuum. 
                                                 Phase   Retentate   Permeate                           Amount in relation to     45%     55%           dry matter           Polyphenol content in   44.4%   15.4%           relation to native           extract                        
The enriching of polyphenols in the retentate proceeds specifically, while even producing a significantly higher content than could be assumed from the mass relationship. In comparison to the initial extract with 31.4% of polyphenols, the content could be increased by 42%.
 
     EXAMPLE 8 
     Liquid-Liquid Extraction 
     100 g native extract—as obtained according to example 7 (retentate phase)—is set up to a 20% dry matter ratio with water. Afterwards, it is intensely mixed 3× with 1/3 volume parts n-butanol in a separating funnel at room temperature. After having rested for 30 minutes, a clear phase separation develops. The n-butanol phase is separated from the aqueous phase. 
     The combined organic phases, as also the water phase, were concentrated to a spissum while removing the solvents with the help of a rotary evaporator. 
                                                 Phase   Organic phase   Water phase                           Amount in relation to     16%     84%           dry matter           Polyphenol in relation to   63.7%   27.6%           native extract                        
The enriching of polyphenols through the organic phase is specific and produces an increase of about 49% in comparison to the initial extract with 31.4% of polyphenols.
 
     EXAMPLE 9 
     Antioxidant Potential 
     ORAC (oxygen radical absorption capacity) is an internationally standardized method in order to define antioxidant potentials. By means of this method, a distinction between hydrophilic and lipophilic parts of the antioxidant effect is possible. For better comparability, the total capacity is given in the unit of the trolox equivalent (TE). The actual procedure is described on www.orac-europe.com. 
     The invented  Cistus  extract (60% polyphenol)=5100 μmol TE/g 
     Aqueous  Cistus  extract (25% polyphenols)=1900 μmol TE/g 
     The invented enriched  Cistus  extract has an outstandingly high ORAC potential, which is higher than the current state-of-the-art by a factor of 2.7. Enriching polyphenolic compounds from Herba  Cistus L . correlates with its antioxidant potential. 
     The concentration of oxygen radicals (superoxide anions) is regarded as measure for oxidative stress in organic tissues. By means of quantifying superoxide anions with a color reduction after stimulating with phorbol-12-myristat-13-acetate (PMA), the following antioxidative capacities have been recorded (test concentration 200 μg/ml) in comparison to an untreated control (0% inactivation): 
     The invented  Cistus  extract (60% polyphenols): 65.0±8.9% 
     Aqueous  Cistus  extract (25% polyphenols): 20.5±9.3% 
     Vitamin C (methodical standard): 96.6%±11.3% 
     The invented  Cistus  extract (60% polyphenols) has an outstandingly high antioxidant capacity for inactivating free oxygen radicals, which is increased by a factor of 3.2 in comparison to current state-of-the-art. 
     EXAMPLE 10 
     Drop Formulation 
     Purified ethanolic extract—as obtained according to example 5—is redissolved into water to a dry matter of 12%. Afterwards, 20% glycerol is added while stirring. Thus, a drop formulation was obtained, which has an improved taste, and a longer endurance. 
     EXAMPLE 11 
     Application as Nasal Drop 
     A drop formulation manufactured according to the aforementioned example 10 was used by a male proband, aged 38, in the course of a violent hay fever sneezing attack. He used it nasally in the form of drops from a pipette. After only a few minutes, his nasal mucosa was no longer swollen and reduced to its normal size. 
     EXAMPLE 12 
     Usage as Nasal Spray 
     A drop formulation manufactured according to the aforementioned example 10 was used by a male proband, aged 56, in the course of a violent hay fever sneezing attack. He used it nasally, nebulized to a mist from a nasal spray. By the spray dose, the sneezing attack was reduced after a few minutes.