DE3720232C2 - Process for the preparation of an extract from Aloe leaves of the plant genus Aloe capensis or Aloe arborescens - Google Patents

Description

The invention relates to a method for producing a Extract of aloe leaves of the plant genus Aloe capensis or aloe arborescens, in which the aloe leaves at low Temperatures or stored in darkness for at least 12 days be a biochemical remodeling in the tissues of the Leaves under formation of biogenic stimulators is initiated and the juice of the leaves is then extracted.

The invention further relates to the use of the after Aloe extract prepared according to the invention, prepared as solution for injection to mobilize the Immune system using s. c. Injection or intramuscular  Injection, and prepared as an emulsion according to the later listed approaches to external application to the hand to mobilize the immune system.

After the school of Prof. Filatow (see reports of the Eastern Europe Institute at the Free University of Berlin, edited by Prof. dr. Max Brandt, Episode 18/1, ed. 1960), aloin is a stimulant and in particular a tissue therapy known after the fresh juice of aloe leaves used for wound healing.

If aloe leaves or aloe extract for about 12 days at low temperature or in darkness, form biogenic stimulators, according to Filatov in the sense a non-specific stimulus therapy used for tissue therapy become.

This tissue therapy in the sense of a non-specific stimulus therapy uses certain active ingredients unclarified Nature, which is present in plant and animal tissues Conservation develop in cold or dark.

The invention has for its object to provide a method of be mentioned in the opening paragraph so that an aloe extract can be produced in the biologically active agents  which contain those acceptors in the central nervous system speak in favor of a mobilization of the Immune system are responsible.

In addition to the new, not listed in Hager's manual Indications of the Aloe extract produced according to the invention regarding the cure of epilepsy and breast cancer, which can be proven by corresponding disease progress reports is the indication to change the leukocyte image stressed in the blood. The leukocytes, white blood cells (nucleated, round corpuscles, larger than the red blood cells), are important elements to effective Ward off infections. You are responsible for that Forträume and "erosion" of undermined tissue components (Pus) as well as bacteria. The leukocytes divide into granulocytes and lymphocytes, with the Lymphocytes are carriers of the immune system.

After applying 9-15 injections of the invention prepared extract of biostimulated aloe capensis (D 2), the leukocyte image in the blood in patients increased from 1800 to 4400 on average.

Thus, the extract prepared according to the invention - from Fresh plant aloe biostimulated - a basic immune repellent  with the widest range while the only material oriented previous Aloe preparations only the Fulfill indication claims, from the Hagers manual are known.

This object is achieved by the method steps according to the characterizing part of patent claim 1

Advantageously, the storage temperature is 2 ° C selected.

It should be emphasized that the HAB rules only for such Apply to drugs whose quality and identity are determined by the European Pharmacopoeia can be determined. This is at aloe in terms of fresh drugs so far after the Literature and the prior art not possible. Much more are in HAB only test criteria for dry drugs of aloe contain. But according to EAB identity and quality can not be determined, so uses due to lack of it Condition also the prescription over the production of drugs after the HAB nothing.

The invention therefore had its own monograph for the Identification of identity and quality of the fresh drug produce.

Sample preparation for liquid chromatographic determination of Aloin in Aloe D2 1. Preparation of the sample solution

Enrichment of aloin is achieved by means of a CH- (cyclohexyl-) pillar carried out.

For conditioning, first with 2 ml of methanol and then 2 ml of water rinsed.

Then exactly 25 ml of Aloe D2 are passed through the column given. After the aqueous phase has been infiltrated, is eluted with 10 ml of methanol. To get a uniform To achieve flow rate of the eluent must light water jet vacuum can be applied. The eluate is concentrated on a rotary evaporator to <1 ml and completely transferred to a 1 ml volumetric flask, which is up to is filled to the mark with H₂O.

2. Preparation of the calibration solution

12.8 mg aloin are dissolved in 100 ml H₂O. This is exactly 0.1 ml into a 100 ml volumetric flask, which is to the mark  is filled up with water. Subsequently, the enrichment process - As described above - performed.

in-process control

• a) Properties:
  Clear, colorless liquid
• b) pH:
  The pH is measured potentiometrically according to Ph.Fur.I with a combination electrode.
  Limit: 7.2-7.4
• c) Removable volume:
  The test of the removable volume is carried out according to the monograph Paren Heralia Ph. Fur.III.
  Limit: 1.05-1.10 ml with 1 ml glass ampoules.
• d) melting level:
  The Abschmelzhöhe is determined with a caliper.
  Limit: 47 ± 2 mm for 1 ml glass ampoules.

1. Development of a procedure for the determination of pharmaceutical grade

It was aimed at both the identity and the salary to standardize with a method. The pharmaceutical grade of aqueous aloe preparations  is checked by a determination of the anthraquinone pattern, the content is indicated as aloin, a 1,8-dihydroxy-anthraquinone, in mg aloin per 1000 ml liquid Preparation.

Usually, such a determination with an HPLC method carried out.

1.1 The development of the process 1.1.1 Compilation of the device configuration

The beginning of the process development consists in the compilation the appropriate device configuration as well as in the selection of the optimal separation ensuring eluent. Experience has shown that the eluent is a mixture of Acetonitrile and 1-hexanesulfonic acid (PIC B-6 reagent, Waters Associates) gives the best results.

To find all relevant anthraquinone derivatives in an acceptable Time elution was a gradient HPLC method elected as in an isocratic process Signals with a high retention time are no longer sufficient Accuracy can be integrated.

The HPLC column was the Kontrosorb 10C18 column (250 × 4 mm). selected.  

For detection has a spectrophotometer with a Standard wavelength of 254 nm as perfectly adequate proven; However, a measurement of the UV maxima of Aloin has shown that the detection optionally also at 295 nm or can also be done at 365 nm.

1.1.2. Optimization of the procedure

The use of the solvent mixtures has proven suitable A: acetonitrile-PIC B-6 (10:90) and B: acetonitrile-PIC B-6 (90:10). The gradient curve is linear from 0% B on 40% B within 35 min.

The aloin peak was identified by first three Samples of the aqueous extract were injected, to which different amounts of aloin were added. The aloin peak eluted at a mean retention time of 28 min.

1.2. Preparation of the calibration solution

The quantitative determination of the aloin content was carried out according to the method of the external standard. This was initially after some preliminary tests to estimate the approximate Aloin content of a calibration solution: 5.85 mg aloin is added to a 100 ml volumetric flask with water (Baker Analyzed HPLC Reagent) and then 10 minute treated in an ultrasonic bath (see chromatogram Attachment 1).  

1.3. Carrying out the measurement

System: HPLC pumps Waters 501
Column: Kontrosorb 10C18, 250 × 4 mm (Kontron analysis)
Eluent A: acetonitrile (Merck LiChrosolv) / PICB-6 (Waters Associates) 10:90
Eluent B: acetonitrile / PIC B-6 90:10
Degree. Course: from 0% B to 40% B in 35 min. (linear increase)
Injection amount: 20 microliters
Flow: 1.0 ml / min.
Detection: Spectrophotometer Uvikon 740 LC (Kontron Analytics)

1.4. sample preparation

The samples of the two aqueous preparations could after Ultrafiltration can be injected directly.

2. Examination results 2.1. Aqueous Aloe extract (see Appendix 2)

The aloin content of the aqueous aloe extract is 33.8 mg / 1000 ml.

2.2. Aqueous buffered mother tincture (see Appendix 2)

The aloin content of the buffered mother tincture is 86.5 mg / 1000 ml.  

Spectra have been generated (see Appendices 3 to 5), which show the biogenic stimulators for the first time:

For the preparation of the starting materials were leaves cut from a single aloe arborescens (aloe capensis). 100 g were immediately squeezed out, centrifuged and washed and measured, 100 g were equivalent to 14 days biostimulated according to the method of the invention, squeezed out, centrifuged and measured and washed. The following changes can be seen:

Aloe arborescens (capensis) without biostimulation (Appendix 3)

• 1. Derivation: (Appendix 4)
  At 250 nm (2) normally formed peak
  at 225 nm (1) hump, ie superposition of 2 closely spaced peaks
• 2. Derivation of aloe arborescens biostimulated (capensis):
  At 250 nm (2) widespread peak (also overlay)
  at 225 nm (1), newly formed peak, ie separation of the closely spaced peaks
• 3. Derivation: (Appendix 5)
  Biostimulated formation of a small new peak at 225 nm (1) and formation of a hump at 250 nm (2).
  Formation of a new substance is thus identified.

Also in the visual range from 400 nm are on Appendix 5 To recognize changes (3 + 4).  

According to the invention, the extract prepared for action on the functional state of the information flow system of the central nervous system by means of s. c. Injection or intramuscular injection.

About the action of the applied according to the invention Aloe extract on changes of the functional State of the central nervous system is in the measure of a broadband effect the regeneration and strengthening of the body's own Defenses - strengthening the immune system z. Against AIDS - given. The broadband effect of aloe extract makes This successfully used against a variety of diseases.

The selected plants must be at least 6 years old, so that the plant tissue is developed far enough to in the process sequences according to the invention a refinement of biogenic stimulators in plant tissue in high molecular weight To allow state in which its mechanism of action the biogenic receptors of the central nervous system equivalent. Are aloe extracts of younger plants mentioned Art applied, in which the refinement of the Biogenic stimulators could not take place, it is only an incentive of some receptors recognizable, not however a broad spectrum in the central nervous system, so that in the Apply a strong burning sensation for 3 minutes in the tissues the living thing occurs. When applying the invention  aloe extract made this burning does not occur on.

The nature of the biogenic stimulators in the invention prepared aloe extract corresponds to one comparatively Hormone, but is in the process of enzymatic action ability, Hormones are destroyed when heated, the biogenic stimulators on the other hand, they remain at high heat, at conversion the substance from liquid to solid and vice versa completely received effectively. The refined biogenic stimulators on the other hand, have kinship with the brain enzyme system. This is due to the action of the refined biogenic Activated stimulators. If through agents the acceptors cause a biochemical remodeling in the central nervous system, so occurs the same biochemical remodeling in the host organ separate and in unfavorable conditions, however allowed to kill, put tissues, the new biological creates active connections, new life regulators. hormones These are not, as hormones are among the currently possible ones Measuring conditions are destroyed. They are other subtle, biologically active substances. They come next the class of heteroglycans, namely those heteroglycans, which one can call DESMONE. But the refined ones Biogenic stimulators are more intense because they are at the homeopathic dilution stage D2 particularly active.  

In the application of the Aloe extract according to the invention was z. B. found that in 27 patients, for general stimulation 20-50 aloe injections have received some changes in the blood were. In 9 patients, the titre was that of anti-A and anti-B agglutinins increased.

Furthermore, it was found that after application aloe extract with the refined biogenic stimulators almost half of the patients had irregular antibodies, which donor erythrocytes of all blood groups agglutinate could.

Irregular antibodies are after Pschyrembel immune antibodies. The fight of the system of the body's defenses supports on antibodies. With the introduction of the invention prepared aloe extract in the pharmacy and medicine It is possible for the first time by the refined biogenic Stimulators in aloe extract a product available to put that antibody formation and thus the immune system strengthens.

By injecting refined biostimulated aloe extract will the intrarenal circulation as well as the Excretory function of the kidneys clearly stimulated. The  Blood flow to the kidney is activated, the speed the glomerulofiltration increases. Furthermore results an improvement of the nitrogen excretion function. Also beneficial is the influence of the refined biogenic Active ingredients on the kidney capsule tissue.

To assess the effect of the invention produced Aloe extract on the functional state of the central nervous system the method of conditional reflexes is used. It can be seen that the use of the invention By means of a delay of the latent period of the reflexive Reactions, increasing the reflex duration, lowering the height which causes conditioned reflexes to their complete failure. From a reinforcement of the inhibition process in the CNS can be spoken.

Of a significant role of the central nervous system in the Mechanism of action of aloe preparations testify the observations the pharmacodynamic changes of aloe preparations with changes in the functional CNS condition.

For the clarification of the mechanism of action of the invention prepared aloe extract is the study of Activity of the brain enzyme system of particular importance.  

Under the influence of the aloe extract can in the cerebrum of the Animals an increase in activity of some oxidation enzymes such Cytochrome oxidase, dehydrase, succinate hydrogase, phosphate dehydrogenase, Glutamate dehydrogenase, succinoxydase to be observed.

The emulsion is for external use only and will applied to the skin like an ointment or a balm.

According to the invention, six subsequent approaches were Aloe emulsion successfully tested:

Aloe emulsion

1st approach

about 40 ml of aloe in H₂O (2 g of aloe to 75 ml H₂O)
5 ml of castor oil
5 g of cetyl alcohol
1 g beeswax
a spatula tip Borax
0.1 ml of eucalyptus oil

2nd approach

75 ml Aloe solution (1.5 g Aloe in 75 ml H₂O)
10 ml of castor oil
10 ml of cetyl alcohol
5 g of beeswax
about 0.1 ml of eucalyptus oil

3rd and 4th approach

75 ml Aloe solution (1 g Aloe in 75 ml H₂O)
10 ml of castor oil
10 ml of cetyl alcohol
about 5 g of Lanalin
about 0.1 ml citric or rose oil
about 0.5 ml of almond oil

5th approach

75 ml Aloe solution (1 g Aloe in 75 ml H₂O)
10 mg castor oil
2 mg of cetyl alcohol
5 g of beeswax
2 g of treetherpolamine
1 g almond oil
about 0.1 ml of lemon oil

6th approach 69.9% by weight a solution prepared in the proportion of 1 part aloe extract and 9 parts purified water 15.0% by weight castor oil 15.0% by weight cetyl alcohol 0.1% by weight eucalyptus oil

Claims (4)

A process for producing an extract of Aloe leaves of the genus Aloe capensis or Aloe arborescens, wherein the aloe leaves are stored at low temperatures or in darkness for at least 12 days, wherein a biochemical modification in the tissues of the leaves to form biogenic stimulators and the juice of the leaves is then extracted, characterized by the following process steps:

• (a) 10 g of fresh aloe leaves freshly cut from the stem of at least six-year-old plants of the said species and briefly dipped after being cut into hot water, which plants must not be watered for 5 days before cutting the leaves, are triturated together with 20 g of a known ascorbate phosphate buffer solution in a mortar to a pulp, whereupon the dry loss of the fresh aloe determined at 97% and based thereon a total of 38.8 g of the previously prepared buffer solution are added to the batch, the amount already withdrawn from the calculated amount, which batch is stored immediately thereafter at temperatures between 2 ° C and 4 ° C for 14 days in simultaneous darkness,
• b) then the macerate is allowed to stand at room temperature for 60 min, squeezed, filtered and the extracted juice stored for 14 days at temperatures between 2 ° C and 4 ° C and simultaneous darkness, and
• c) the thus-obtained mother tincture of biostimulated aloe is diluted with 19 parts of buffer solution to give the potency D2.

2. The method according to claim 1, characterized in that as storage temperature 2 ° C is selected.   3. Use of the method according to claim 1 and 2 prepared extract, prepared as a solution for injection to mobilize the immune system by means of s. c. injection or intramuscular injection.   4. Use of the extract prepared by the process according to claims 1 and 2, prepared as an emulsion according to any of the given external topical formulations for the mobilization of the immune system. 1st approach 40 ml Aloe in H₂O, (2 g Aloe to 75 ml H₂O)
5 ml of castor oil
5 g of cetyl alcohol
1 g beeswax
a spatula tip Borax
0.1 ml of eucalyptus oil2. Mixture 75 ml Aloe solution, (1.5 g Aloe in 75 ml H₂O)
10 ml of castor oil
10 ml of cetyl alcohol
5 g of beeswax
about 0.1 ml of eucalyptus oil3. Mixture 75 ml Aloe solution, (1 g Aloe in 75 ml H₂O)
10 ml of castor oil
10 ml of cetyl alcohol
about 5 g of Lanalin
about 0.1 ml citric or rose oil
about 0.5 ml of almond oil4. Mixture 75 ml Aloe solution, (1 g Aloe in 75 ml H₂O)
10 mg castor oil
2 mg of cetyl alcohol
5 g of beeswax
2 g of treetherpolamine
1 g almond oil
about 0.1 ml of lemon oil 5th approach 69.9% by weight a solution prepared in the proportion of 1 part aloe extract and 9 parts purified water 15.0% by weight castor oil 15.0% by weight cetyl alcohol 0.1% by weight eucalyptus oil