FR2780651A1 - METHOD FOR ACTIVATING AN INACTIVE AND VERY LOW CONCENTRATION SOLUTION OF A BIOLOGICAL AND / OR CHEMICAL SUBSDETERMINE IN A SOLVENT - Google Patents

Abstract

The invention relates to a method for activating an inactive and very low concentration solution of a determined biological and / or chemical substance. The method comprises the step of subjecting said solution to a mechanical excitation field, in particular created by a vortex. .

Description

Method for activating an inactive and very low concentration solution

  of a specific biological and / or chemical substance

in a solvent.

  The present invention relates to a method for activating an inactive and very low concentration solution of a determined biological and / or chemical substance in a solvent. The present invention

  also relates to the applications of said activation method.

  The term "very low concentration" denotes concentrations

  between 10 - 6 and zero moles per liter (M).

  Methods for preparing a "highly diluted" solution are known by successive dilution and stirring. One of the preparation methods traditionally used in homeopathy (Hannemahn's method) consists, starting from a relatively concentrated solution made using a mother tincture (of concentration greater than 10 - 6 M), to carry out a dilution by a factor of 10 or 100, then mechanical agitation (called "dynamization"). It is noted after each operation that the solution has remained active in the sense that it triggers a reaction within

of a sensitive biological system.

  As examples of solutions which have been prepared by the method below

  above described, we can cite all the homeopathic preparations. By way of examples of sensitive biological systems making it possible to test the active nature of such solutions, there may be mentioned: the isolated guinea pig heart (Langendorff experiment) or the skin test carried out on the skin of a

guinea pig or a living rabbit.

  The need to start with an active solution before diluting and then stirring seemed essential to obtain active diluted solutions. Dilutions carried out up to 10 '6 M and beyond, without implementing this successive stirring and dilution process, have not so far made it possible to obtain active solutions. It is not without interest either to recall that the stirring and dilution technique implemented by Hannemann has been extrapolated without it being possible to show the virtues of such high dilutions beyond factor 5 until now. CH. However, the inventors, who are known for their work on high dilutions (Nature 1988: "Degranulation of human Basophils triggered by a high dilution solution of anti-IgE antibodies"), have surprisingly found that it was possible to obtain an active solution from an inactive and very low concentration solution of a determined biological and / or chemical substance in a solvent. They have shown that a solution at very low concentration, the activity of which does not exist at the outset, can be made active by methods which are particularly simple to implement. Thus, they designed a process which makes it possible to make active solutions at very low concentration without it being necessary to prepare them beforehand by the technique

  traditional successive dilutions and agitations.

  They have thus solved a problem whose industrial implications are considerable. Indeed: - it is now possible to detect certain biological and / or chemical substances in solution at very low concentration in a solvent, - it is now possible to design and produce drugs using specific biological and / or chemical substances in solution at very low concentration in a solvent, - it is now possible to control the production of products to

  very low concentration including homeopathic products.

  The present invention therefore relates to a method for activating a solution at a very low concentration of a determined biological and / or chemical substance in a solvent. Said method comprises the step of placing said solution in a mechanical excitation field. The mechanical excitation field could be created, for example, by a shock wave propagating in the solution, by ultrasound or sound waves diffused in the solution, by vibrations transmitted by the container containing the solution. Preferably, the mechanical excitation field results from violent agitation, in particular obtained by means of a Vortex-type agitator consisting of a disc which rotates rapidly until a vortex is formed in the column of liquid in restlessness course. Advantageously, said solution is subjected to a field

  mechanical excitation for at least 15 seconds.

  As examples of substances in solution which have been activated by the method described above, there may be mentioned: arnica, ovalbumin,

  acetylcholine, calcium ionophore.

  The method according to the invention is of interest only when the concentration of said determined substance in said solution is less than 10-6 moles per liter. Indeed above 106 moles per liter the solution is already active (traditional pharmacology). Advantageously, the concentration of said substance determined in said solution is within a range of less than 106 moles per liter and greater than

1016 moles per liter.

  Preferably also said solvent contains at least 5% water. It has indeed appeared that below a certain proportion of water in the solvent the solution subjected to a mechanical excitation field remains

inactive.

  It has also been found that a percentage of alcohol of at least 2% in the solvent has the effect that the solution remains active for a longer period. The method according to the invention further comprises the step of controlling the active state of said solution by implementing an experimental protocol identical or similar to that which would be used to account for the presence of said determined substance. in an environment that would contain it. As an example of a sensitive biological system making it possible to test the active nature of the solutions of the following determined substances: calcium ionophore, acetylcholine, histamine, ovalbumin (in animals made sensitive), arnica, bradykinin, anti-IgE antibodies, may cite: the Langendorff experience (isolated guinea pig heart) as well as the skin test carried out on the skin of a living guinea pig or rabbit. Thus, for example, to control the active state of the acetylcholine solution, it is verified that an injection of the latter, in particular under the skin of a guinea pig,

causes skin reactions.

  The present invention also relates to the application of the method to the

  detection of a specific substance, diluted in very low concentration.

  As soon as the solution is activated, it is possible to carry out identification tests by implementing an experimental protocol identical or similar to that which would be used to account for the presence of said determined substance. in an environment that would contain it. As an example of the determined substances that one can

  detect in solution, the following substances may be mentioned: calcium-

  ionophore, caffeine, nicotine, bacterial toxins and endotoxins; and the following identification tests: Langendorff's experience (isolated guinea pig heart) as well as the skin test performed on guinea pig skin or

living rabbit.

  The present invention also relates to the application of the process to the production of medicaments using biological and / or chemical substances at very low concentration. Indeed, as soon as it is possible to prepare active solutions with a very low concentration of determined biological and / or chemical substances, new

  therapeutic applications of these substances are becoming possible.

  Examples of drugs which can thus be produced include the following drugs: coronary vasodilators (ex trinitroglycerin), beta-blocker (propranolol), caffeine, nicotine. The present invention also relates to the application of the method to the control of the production of products at very low concentration, in particular homeopathic products. One of the problems to be solved when manufacturing homeopathic products is that of the production control of successive dilutions. The method according to the invention makes it possible to test the activity of homeopathic products during the various phases of their manufacturing process. By way of example of homeopathic products, the production of which can thus be controlled, the following drugs may be mentioned: arnica 5 to 30 CH, histaminum 5 to 30 CH,

  acetylcolinum 5 to 30 CH, apis mellifica 5 to 30 CH.

  Other characteristics and advantages of the invention will become apparent on

  reading of the description of variant embodiments of the invention, given

  by way of an indicative and nonlimiting example, as well as on reading the examples of experiments which have made it possible to validate the activation process, object of the present invention, and which refer to the appended drawings in which: - the figure 1 shows a perspective view of an alternative embodiment

of the agitation system.

  - Figure 2 shows a perspective view of a system for

  test the activity of the solution (Langendorff experiment).

  - Figure 3 shows an image of the skin of a guinea pig We will now describe Figure 1 which shows a perspective view of an alternative embodiment of the stirring system. A rubber roller 2 is pivotally mounted about a vertical axis. The rubber roller is rotated about the vertical axis by an electric motor (not shown), located inside the housing 3. The electric motor is powered by a cable 10. The speed of rotation of the roller is

  controlled by a potentiometer 4.

  Tube 1 contains solution 6 of acetylcholine at the concentration of lpM, to be shaken violently. The operator 5 maintains the lower end 7 of the tube applied against the roller 2 by pressing on the upper part 8 of the tube 1. The lower end 7 of the tube 1 describes a circle 9. As a result, a vortex occurs within solution 6. This vortex

  shake and mix the solution violently.

  By way of example, we will now describe, with reference to FIG. 2, the test carried out from an isolated guinea pig heart perfused, known since 1897 (under the name of "Langendorff experiment") and described in books on traditional pharmacology, in particular in "Animal experimentation in cardiology" - Medicine - Science Inserm - Flammarion, Bernard SWYNGHEDAUW, Chapter 3.1 P. 81 "Isolated organ: isolated heart according to Langendorff". The fraction collector collects the tubes at the rate of one tube per minute and thus measures the flow rate of the guinea pig heart minute by minute. Here are the results from the experiments carried out with the following substances, after they have vortexed (agitated) as it was described with reference to FIG. 1, in a 15 ml tube containing 10 nml of solution: - for the first , a mixture of acetate-choline (AC) lpM (sodium acetate 1 pM + choline chloride 1 pM) vortexed for 15 seconds, - for the second, acetylcholine (ACh) lpM vortexed for 15 seconds, - for the third, acetylcholine (ACh) lpM vortexed for 5 seconds - for the third, acetylcholine (ACh) lpM vortexed for 2 seconds - for the third, acetylcholine (ACh) vortexed for 1 second La Buffer solution had the following composition: Ca 2+ 2mM, NaHCO3 mM.

  The solvent used in all five cases was water.

  The table below indicates (in ml) the amount of buffer solution

  recovered in the collecting tubes over time.

  Exp 1 Exp 2 Exp 3 Exp 4 Exp 5 Time acetylcholine acetylcholine acetylcholine acetylcholine acetylcholine (minutes) choline (ACh) (ACh) (ACh) (ACh) (A-C) Vortex 15s Vortex 15s Vortex 5s Vortex 2s Without Vortex

1 3.7 2.9 3.5 3.7 3.6

2 3.8 3.0 3.6 3.8 3.7

3 3.7 3.0 3.6 3.7 3.7

4 3.8 2.9 3.5 3.7 3.6

3.7 2.8 3.3 3.8 3.8

6 3.7 3.3 3.8 3.6 3.8

7 3.6 3.3 3.8 3.5 3.7

8 3.6 3.4 3.7 3.5 3.6

9 3.7 3.3 3.6 3.6 3.6

3.8 3.2 3.5 3.5 3.7

1 3.7 3.0 3.4 3.5 3.7

12 3.7 2.9 3.5 3.6 3.8

13 3.6 3.1 3.4 3.5 3.7

14 3.7 3.0 3.4 3.5 3.6

3.7 2.9 3.4 3.6 3.6

3.6 2.8 3.3 3.5 3.6

3.7 2.8 3.2 3.5 3.7

3.6 2.8 3.2 3.5 3.6

  This table highlights several things: a) in the experimental protocol the solutions are tested before being vortexed to verify that they are inactive. The results of Experiment 5 illustrate one of these tests. The solution is inactive before being vortexed. In fact, the variation in flow rate of 0.2 ml / min (min = 3.6; max = 3.8) corresponds to measurement uncertainties and to normal variations in flow rate in the biological system, which is the isolated guinea pig heart perfused. b) the comparison of the results of experiment 1 with experiment 2 shows that the action of agitation is not sufficient in itself: still it is necessary that molecules to which the biological system is sensitive are present. Indeed a product very close to acetylcholine, acetate-choline, prepared under the same conditions as acetylcholine,

  does not trigger a reaction (Exp 1).

  c) the series of experiments 2, 3, and 4 was carried out on the same guinea pig core and shows the influence of the agitation time using the Vortex on the activity produced in the substance. Thus for a 2 second agitation we obtain a maximum variation of flow rate of 0.3 ml / min (min = 3.5; max = 3.8) while for a 15 second agitation we obtain a maximum variation of flow rate of 0.6 ml / min (min = 2.8; max = 3.4) As further examples, here are the results from the experiments carried out with acetylcholine lpM substance, vortexed for 15 seconds for different ethanol contents in the solvent. The

  experiments were carried out 9 days after stirring the solution.

  % variation in flow rate variation in ethanol flow rate coronary artery coronary artery (in ml / min) (based on average flow)

1% 0.6 10%

2% 0.7 14%

% 0.9 18%

% 1.1 26%

  This table shows that the ethanol content favors the

  conservation of activity in water.

  We will now describe with reference to Figure 3 the skin test in guinea pigs We use a living guinea pig, which is injected venously with a blue dye (Evans blue) which binds to the albumin in the blood. Albumin does not leave the vessels, unless there is inflammation, therefore vasodilation and permeation of the vessels, the typical example of such a reaction in

the man being hives.

  The test is carried out by injecting 0.1 ml of the solution under the skin of the thus prepared animal, the activity of which should be checked. The diameter of the blue spots that appeared around the injection points is then measured. To do this, we scan the skin, then save the bitmap file. Then we

  measures the pixel size of the blue spots due to the reaction.

  The numbers 3, 4, 10, 11 which appear in the first column of the table

  below correspond to the references of figure 3.

  N Solution injected Total Pixels in thousands 3 vortexed lpM ACh 1,949 4 non vortexed lpM ACh 43 vortexed lpM AC 25 11 ACh 1 vM vortexed 1,154 - Injection number 3 shows that the very low concentration vortexed solution (lpM) neuromediator acetylcholine (ACh) triggers a significant skin reaction (1,949,103 pixels) compared to the same non-vortexed solution which does not trigger a reaction as shown

  injection number 4 (43,103 pixels).

  - The comparison of injection number 3 (1,949,103 pixels) and injection number 11 (1,154,103 pixels) shows that the activity of a solution at very low vortexed concentration is quite comparable to that of '' a non-vortexed solution with higher concentration (lgMI, usual concentration in traditional pharmacology) - Injection number 10 is carried out with a vortexed picomolar solution (lpM) of a product close to acetylcholine but inactive: the acetate / choline (AC). This injection shows that the skin reaction of the guinea pig is very specific to the nature of the substance in solution because this acetate / choline solution is vortexed under the same conditions as

  injection number 1 has no effect (25,103 pixels).

Claims (12)

Claims   1. A method for activating an inactive and very low concentration solution of a determined biological and / or chemical substance in a solvent, said method comprising the step: - of placing said solution in a field of mechanical excitation. 2. Method according to claim 1, said method comprising the step: - of subjecting said solution to stirring to create said field mechanical excitement.   3. Method according to any one of claims 1 or 2, such as   concentration of said substance determined in said solution is less than 10 'moles per liter.   4. Method according to any one of claims 1 to 3, such as   concentration of said substance determined in said solution is within a range of less than 10 -6 moles per liter and greater than -16 moles per liter,   5. Method according to any one of claims 1 to 4, such that said solvent contains at least 5% water.   6. Method according to any one of claims 1 to 5, such that said   solvent contains at least 2% alcohol,   (so that the solution remains active for a longer period).   7. Method according to any one of claims 1 to 6, said method   comprising the step: - of subjecting said solution to a mechanical excitation field for at least 15 seconds.   8. Method according to any one of claims 1 to 7 comprising   in addition the step: - of controlling the active state of said solution by implementing an experimental protocol identical or similar to that which one would implement to account for the presence of said determined substance in an environment that would contain it.   9. The method of claim 8 such that to control the active state of said solution it is verified that an injection thereof causes skin reactions   10. Application of the method according to any one of claims 1 to   9 upon detection of a specific substance, diluted to a very low concentration.   11. Application of the method according to any one of claims 1 to 9 to drug production.   12. Application of the method according to any one of claims 1 to   9 to control the production of very low concentration products,   including homeopathic products.