WO2008056388A1

WO2008056388A1 - Drug based on ozonized terpenes or terpene-derivatives, processes for producing it and uses thereof

Info

Publication number: WO2008056388A1
Application number: PCT/IT2007/000761
Authority: WO
Inventors: Flavio Torresin; Gualtiero Valeri
Original assignee: Istituto Fitofarmaceutico Euganeo S.R.L.; Manni, Ruggero
Priority date: 2006-11-06
Filing date: 2007-10-30
Publication date: 2008-05-15
Also published as: WO2008056388A8; ITFE20060032A1

Abstract

A drug for treating pathologies manifesting as inflammatory processes or as microbic infections, characterised by consisting of terpene ozonide or a derivative thereof pure or contained at high concentration in a natural substance.

Description

DRUG BASED ON OZONIZED TERPENES OR TERPENE-DERIVATIVES, PROCESSES FOR PRODUCING ITAND USES THEREOF

The present invention relates to a drug based on ozonized terpene or terpene-derivative for treating pathologies manifesting as inflammatory processes or as microbic infections, a process for producing the drug, and the use of the drug.

Ozone is an allotropic form of oxygen, formed from triatomic molecules instead of biatomic, as is ordinary oxygen.

A particularly important reaction with organic substances is that of addition at the double bonds, with formation of cyclic groups - (molozonides and ozonides (also defined in the literature as trioxolanes).

The reaction rate of pure terpenes with ozone is very high, even of the order of 10⁶ moles/l.s (moles of ozone per litre of terpene per second), and the reaction is markedly exothermic, making its control difficult.

Ozonides are known to split to form two aldehydes if subjected to the action of water and of a reducing agent; alternatively they can split to form an aldehyde and a carboxylic acid. It is apparent that the decomposition kinetics of the ozonide derivatives is accelerated by the presence of water.

Although various studies have been carried out on the medical application of free ozone and on the pharmacological applications of ozonized vegetable oils, and occasionally on other ozonized unsaturated substances, the following electrochemical or biophysical aspects have never been observed: 1. Ozonized compounds are characterised by high electrochemical electrode potentials when they come into contact with tissues, determining potentials and currents which logically cannot be ignored at cell level.

2. The theory of asepsis is at least partially reinterpreted in accordance with more modern physico-chemical schemes; each type of microorganism has its own life field which, when represented on a four-axis Cartesian diagram, become the electrochemical potential, hydrogen ion activity, osmotic pressure and temperature. Each microorganism has its own precise life domain within this four-dimensional space, outside which it enters a sluggish state or dies. Louis Pasteur had already observed the importance of the pH factor for fermentations, and of the temperature factor for the development of anthrax in chickens. Studies on the growth of marine organism colonies are also known (e.g. Molgula) based on salt concentration and temperature. Pasteur had already noted indirectly, i.e. on the basis of electrochemical potential, the difference in the development of yeast colonies and their metabolism in an aerobic or anaerobic environment.

These considerations could be extended to the particular case of the role of electrochemical potentials in the organism and to a general physiological model based on the theory of the chemical potential, which would open a third branch of physiology: after the physiomechanical and physiochemical study of the phenomena, a study based on a physioenergy definition which is not limited to considering the biochemical reactions in play, but also embraces the thermodynamic aspect. This basis leads to the formulation of a more general interpretation of the pharmacological action of ozone derivatives, i.e. an electrochemical/thermochemical interpretation in terms of chemical potentials rather than vertically considering their organic chemical aspects.

On this interpretive basis it can be deduced that ozonides act by restoring the ionic electrical membrane potential of the cells of altered tissues. Ozonides also tend to decompose to form free radicals. They are able to recombine with the free radicals generated by metabolic activity, to eliminate them. This may appear unusual, as the eliminating action of free radicals is often associated with reducing agents such as ascorbic acid; however oxidant species can sometimes act as reducing agents towards stronger oxidants (it depends on the respective oxido-reduction potential: a species can hence be reducing towards a stronger oxidant, but oxidising towards a more energetic reducer), which is why hydrogen peroxide is used as a reducing agent in certain analytical chemistry reactions.

With regard to viruses, it is known that ozonides block the viral receptors and kill the virus-infected cells, hence halting their propagation.

Ozonides are also known to also demonstrate good activity against protozoa. They are also active against nematoids.

These considerations give a more complete interpretive idea of the width of the pharmacological action spectrum of these compounds. The peroxide number gives a measure of the electrochemical potential which the obtained compound is able to generate, while its molecular weight gives a measure of its diffusion rate through tissues (the lower the molecular weight, the higher the rate with which the compound diffuses through tissues), hence the ability to reach more or less deep regions by diffusion. Terpenes are unsaturated hydrocarbons (considered multiples of the isoprene unit, hence the term isoprenoidi, included at the biochemical level in the lipid class) which, as such, have the ability to add ozone to the double bond, to form the corresponding ozonide; it has been found that under normal environmental conditions in an anhydrous atmosphere free of reducing agents, these compounds are reasonably stable and survive for long periods. Such ozonides are able to behave as liposoluble oxidants and develop free oxygen, terpenoids are compounds in which a terpene hydrogen atom is substituted by a functional group, and for ozonization purposes behave similarly to terpenes. For ozonization purposes the mono terpenes are more interesting because of their low molecular weight and hence their greater ability to diffuse through tissues; they fall within the class of terpenes, sesquiterpenes, diterpenes, triterpenes, tetraterpenes, polyterpenes. Sesquiterpenes and diterpenes can also be considered interesting for ozonization purposes because of their low molecular weight.

Terpenes and terpenoids are often contained in large quantities in essential oils, which in addition also contain other compounds able to form ozonides following ozonization.

Because of their very low molecular weight compared with ozonized unsaturated triglycerides, ozonized terpenes present an excellent capacity for diffusing through tissues and have a much higher affinity towards water, so increasing their reactivity in biological matrices.

Moreover, terpene and terpenoid ozonides do not present the toxicity which many respective terpenes and terpenoids possess.

It is inferred that the stability of these ozonides with time is related to the length of the aliphatic chain of which the original double bond formed part, even if the oxidizing power becomes gradually less intense. The pharmacological activity of various ozonized organic substances has been known for many years.

Because of their aforestated low molecular weight and their higher reactivity, ozonized terpenes present further new pharmacological properties or present them to a much greater degree than ozonized oleic acid, especially a very high anti-inflammatory activity by percutaneous application. The biochemical characteristics of these substances also mean that they are better accepted than any ozonized unsaturated hydrocarbon (attempts made by other experimenters in the past), in that they are easily metabolised by the enzymatic system of the human and animal organism.

It is known to ozonize essential oils in aqueous emulsion to obtain so- called activated or peroxidated essential oils, the percutaneous absorption of which is described as much greater than essential oils as such. In this respect, it should be noted that in addition to obtaining ozonides or peroxides, there is decomposition and oxidation of the unsaturated compounds with formation of carboxylic acids, because of which there is some reason to doubt the effectiveness of this process for the purpose of obtaining pharmacologically active substances.

According to the invention, these drawbacks are eliminated by a drug for treating pathologies manifesting as inflammatory processes or as microbic infections, as claimed in claim 1.

The invention also relates to a process for preparing the drug, as claimed in claim 2.

The present invention is further clarified in its general formulation with reference to the ensuing description. The process of the invention consists of treating terpenes, terpenoids or essential oils with a high content of these latter, in mixture with fixed oils, Vaseline oil or apolar solvents with a stream of ozone in the form of an ozone/oxygen or ozone/air or ozone/oxygen/nitrogen mixture or ozone diluted in one or more inert gases. An essential characteristic is that the solvents used do not react, or only moderately react, with ozone. These solvents must also be compatible with the use for which the ozonization product is intended.

As a non-limiting example, to suitably dilute the terpenes, terpenoids or essential oils the following can be used: Vaseline oil, vegetable oils extracted from plants or seeds (such as olive oil, Jojoba oil, or any other oil liquid at ambient temperature or close thereto), oleic acid and its esters.

If the terpenes, terpenoids or essential oils were not diluted in this manner, the reaction would be excessively violent and uncontrollable, such mixtures having proved excellent for targeting the obtained ozonides in the uses for which they are intended. Unsaturated vegetable oils, oleic acid and its esters show excellent action synergies when ozonized together with terpenes, terpenoids and essential oils. It is not possible or at least practical to operate on ozonides in the pure state, in particular monoterpenes, because of their instability: limonene ozonide or myrcene ozonide for example tend to ignite spontaneously.

It has been found in practice convenient to use for ozonization a solution of the terpene, terpenoid or essential oil up to 20% in a fixed oil, in

Vaseline oil or in oleic acid and its esters, this concentration also proving suitable for the subsequent pharmacological or cosmetic use of the product obtained. On ozonizing essential oils of high terpene content (for example lemon essential oil, sweet orange essential oil, turpentine essence) dissolved in

Vaseline oil, precipitation of a substance similar to an oleoresin of yellow-gold colour was noted, insoluble in water but soluble in carbon tetrachloride, the exact chemical nature of which has not yet been determined.

As stated, if ozonizing a mixture of a terpene, terpenoid or essential oil with for example oleic acid or olive oil, there is simultaneous ozonization of the terpenes present and of the double bonds of the unsaturated fatty acids present, these compounds showing a synergic action in therapeutic applications, they having different molecular weights and solubilities, hence different rates of absorption by tissues, for example in topical use.

The use of terpenes or essential oils for ozonization purposes is hence important using, as solvents for them, liquid oils or waxes of vegetable or animal origin, or substances obtained therefrom which are liquid at or near ambient temperature, such as oleic acid. Given the ordinary presence of hydrocarbons in vegetable or animal organisms, specifically joined to fats, the use of mineral oils of pharmaceutical type (for example Vaseline oil) can be considered in line with that specified above.

The two most important merits of ozonized terpenes, terpenoids or essential oils compared with ozonized vegetable or animal oils are the following:

- a molecular weight generally equal to about one fifth or one sixth, giving greater ease of diffusion through tissues;

- a higher specific active oxygen content, i.e. a greater ratio of active oxygen/moles of substance. Moreover, by treating the terpenes, terpenoids or essential oils in the anhydrous state, the ozonides which form remain stable because of the impossibility of becoming hydrolysed by the action of the water.

The action of ozone on the solution of terpenes, terpenoids or essential oils can last for a time of greater or lesser length, the degree of ozonization obtained being able to be evaluated on the basis of known parameters, of which density is particularly useful at the industrial production level.

At the operative level, the procedure consisted of bringing a stream of ozonized gas (for example an ozone/oxygen or an ozone/air mixture) into contact with the terpene, terpenoid or essential oil solution, using a suitable contact device (as non-limiting example, a bubbler of Dreschel bottle type, a plate column or a packed column) and maintaining the temperature around

20⁰C (not greater than 30⁰C), as the reaction is exothermic. In this respect, at higher temperature the ozone decomposes to form nascent oxygen, which favours the formation of epoxides (potentially toxic and irritant, hence its concentration in the product must be limited as much as possible). High temperatures also favour decomposition of the ozonides of the said compounds; in addition, even large losses of the terpene component occur by volatility. In contrast, if operating at lower temperatures, the solution viscosity increases, particularly if in oils, and contact between the gas and liquid becomes less efficient; a temperature of 10-15⁰C could usefully be used if the viscosity of the solvent used permits it, this reducing the terpene component losses by volatility. The degree of ozonization was determined by monitoring density, as this increases with the degree of ozonization, with the reaction being interrupted when the required density was reached. Hence the required degree of ozonization of the terpene compounds can be achieved, including their complete transformation into the corresponding ozonides.

Usable terpenes and terpenoids include, as non-limiting examples, limonene, α-pinene, β-pinene. terpinene, myrcene, geraniol, neral, terpineol.

Usable essential oils include, as non-limiting examples, lemon essential oil, orange essential oil, turpentine essential oil, hyssop essential oil, nutmeg essential oil, rosemary essential oil, olibanus essential oil.

Usable solvents for terpenes and essential oils include, as non-limiting examples, Vaseline oil, oleic acid, oleic acid esters, jojoba oil, olive oil.

It should be noted that essential oils contain compounds such as eugenol, anetol or cinnamic aldehyde which, when ozonized, develop pharmacological activity similar to that of ozonized terpenes or terpenoids.

The products can be used as such or formulated, by way of non- limiting example, in oils, ointments or pomades, oleogels, or enclosed in capsules or pearls for internal use, pure or mixed with excipients (for example adsorbed in silica, fossil flour, etc.), microencapsulated or reduced to globules. For large surfaces or where it is not easy to apply creams or gels, or as a personal deodorant, the product can be used in spray form. In all cases the composition of the preparations must exclude water.

It has been found possible to improve the pharmacological effects of the products obtained by mixing with them, in suitable manner, medicinal plant extracts, essential oils or other active principles which cannot be oxidized by the formed ozonides or decompose them. On the basis of theoretical and practical studies, the terpenes ozonized and targeted as stated present a pharmacological activity in the following cases, in addition to those already known from the aforestated state of the art:

1. Rheumatic attacks

2. Arthrosis 3. Painkiller

4. Keratitis

5. Keloid scars

6. Hematomas

7. Epidermophytosis 8. Epidermophytosis of the foot

9. Senile cutaneous hyperpigmentation phenomena

10. Infected wounds

11. Necrotic wounds

12. Fistulas 13. Chilblains

14. Swellings from reduced venous-lymphatic circulation

15. Herpes genitalis

16. Herpes simplex

17. Skin blemishes 18. Root canal infections

19. Dental hyperesthesia

20. Indurative hypodermatitis

21. Nematocide

22. Normalizing intestinal function 23. Onychomycosis

24. Chronic external otitis 25. Decubitus ulcer

26. Athlete's foot

27. Prevention and treatment of dairy cow teat infections

28. Various types of rhinitis 29. Stretch marks

30. Treatment of surgical wounds

31. Treatment of warts

32. Treatment of infections from mites

33. Gastroduodenal ulcer 34. Ulceration of the lower limbs

35. Vasodilator

Even in the case of pathologies already cited in the aforestated state of the art, the ozonization products obtained as stated show a more complete, ready and effective action. Another application for these products in the cosmetics field is in formulating personal deodorants, which do not require the presence of aluminium salts or antiperspirant products; in this respect, the bacterial action of ozonized terpenes inhibits the bacterial growth which leads to unpleasant odours, and moreover those chemical species characterised by unpleasant odours tend to be oxidized and destroyed by ozonized terpenes and terpenoids.

In practical application, whether for preparative or therapeutic purposes, terpene, terpenoid or essential oil solutions to a maximum of 20% in natural fixed oils (for example olive oil), liquid natural waxes (for example Jojoba oil) or substances deriving therefrom which are liquid at ambient or close to ambient temperature (for example oleic acid or its esters) are easy to handle and show excellent therapeutic effectiveness. Hence for ozonization it is not necessary to use particularly low temperatures to obtain a stable and therapeutically effective product, nor is there any danger of the temperature rising uncontrollably, and should the temperature rise excessively because of too rapid ozone feed or insufficient cooling, the reaction mixture remains perfectly stable.

By way of non-limiting example, the following represent some of the more interesting cases of ozonized solutions of essential oils in oleic acid formulated in oils, ointments, pomades or oleogels. EXAMPLE 1

This case relates to a patient aged 92 years suffering from deforming arthritis at the wrists for ten years; after a few days of treatment the pain passed.

EXAMPLE 2 This case relates to a patient aged 54 years suffering from lumbar arthrosis, in pain for twenty days; after two days of application the pain passed.

EXAMPLE 3

This case relates to a patient aged 62 years suffering from arthrosis of the shoulder, with lasting pain for two years; after initial applications the pain decreased, with total pain disappearance as the applications continued.

EXAMPLE 4

Female patient aged 34 years suffering from initial cervical arthrosis with intermittent pain; after initial applications the pain disappeared. EXAMPLE 5 Female patient aged 76 years with traumatic event at the right knee with very evident swelling; noticed reduction and disappearance of the pain after applying the product.

EXAMPLE 6 Male patient aged 48 years with severe ankle sprain; noticed evident reduction of the pain from initial application, then after a week of application, although maintaining normal walking, the pain had totally disappeared, allowing continuation of normal work.

EXAMPLE 7 Female patient aged 53 years with acute L5S1 herniated disc; the pain progressively disappeared after ten days of treatment.

EXAMPLE 8

Female patient aged 74 years suffering from arthritis in the hands and feet, intense pains in the knees and shoulders; the patient applied the product every evening with light massaging of the parts concerned; after three days of application she was able to again move the hands previously blocked by the arthritis; after six weeks of application she was again able to write and manipulate delicate or heavy objects, while the pain in the shoulders, feet and knees had disappeared or nearly disappeared. EXAMPLE 9

Male patient aged 46 years with consequences of a surgical operation in the knee under arthroscopy. Fifteen days after the operation the surgical wounds, although healed, were still very sensitive to pressure and the knee appeared fairly inflamed. After two days of applying a preparation composed of 8 parts of ozonized oleic acid and 2 parts of ozonized turpentine twice a day, the swelling had disappeared and the surgical wounds had lost the painfulness previously felt by the patient.

After two weeks of applying the preparation the situation appeared perfectly normalized and the treatment was able to be suspended.

Claims

C L A I M S

1. A drug for treating pathologies manifesting as inflammatory processes or as microbic infections, characterised by consisting of terpene ozonide or a derivative thereof which is pure or contained at high concentration in a natural substance.

2. A process for preparing the drug claimed in claim 1 , characterised by dissolving a terpene, or a derivative thereof pure or contained at high concentration in a natural substance, in an apolar solvent which is inert or little reactive towards ozone, and then subjecting it to ozonization at a temperature less than 30°C.

3. A process as claimed in claim 2, characterised by operating at a temperature between 10°C and 20⁰C.

4. A process as claimed in claim 2, characterised by using a bubbler.

5. A process as claimed in claim 2, characterised by using a plate column.

6. A process as claimed in claim 2, characterised by using a packed column.

7. A process as claimed in claim 2, characterised by effecting ozonization with an ozone/oxygen mixture.

8. A process as claimed in claim 2, characterised by effecting ozonization with an ozone/oxygen mixture at a temperature between 10⁰C and 20⁰C.

9. A process as claimed in claim 2, characterised by effecting ozonization with an ozone/inert gas mixture at a temperature between 10°C and 20⁰C.

10. A process as claimed in claim 2, characterised by effecting ozonization with an ozone/oxygen/nitrogen mixture at a temperature between 10⁰C and 2O⁰C.

11. A process as claimed in claim 2, characterised by effecting ozonization with an ozone/oxygen mixture.

12. A process as claimed in claim 1 , characterised by effecting ozonization of terpene or of a derivative thereof pure or contained in high concentration in a natural substance, by contact between it, dissolved in a fixed vegetable oil, and a gaseous mixture containing ozone at a temperature between 10⁰C and 20°C.

13. A process as claimed in claim 1 , characterised by effecting ozonization of terpene or of a derivative thereof pure or contained in high concentration in a natural substance, by contact between it, dissolved in an animal oil, and a gaseous mixture containing ozone at a temperature between 10°C and 20°C.

14. A process as claimed in claim 1 , characterised by effecting ozonization of terpene or of a derivative thereof pure or contained in high concentration in a natural substance, by contact between it, dissolved in vaseline oil, and a gaseous mixture containing ozone at a temperature between 10°C and 20⁰C.

15. A process as claimed in claim 1 , characterised by effecting ozonization of terpene or of a derivative thereof pure or contained in high concentration in a natural substance, by contact between it, dissolved in a liquid wax, and a gaseous mixture containing ozone at a temperature between 1O⁰C and 20⁰C.

16. A process as claimed in claim 2, characterised by effecting ozonization of terpene or of a derivative thereof pure or contained in high concentration in a natural substance, by contact between it, dissolved in Jojoba oil, and a gaseous mixture containing ozone at a temperature between 10⁰C and 20⁰C.

17. A process as claimed in claim 2, characterised by effecting ozonization of terpene or of a derivative thereof pure or contained in high concentration in a natural substance, by contact between it, dissolved in olive oil, and a gaseous mixture containing ozone at a temperature between 10°C and 20°C.

18. A process as claimed in claim 2, characterised by effecting ozonization of terpene or of a derivative thereof pure or contained in high concentration in a natural substance, by contact between it, dissolved in oleic acid, and a gaseous mixture containing ozone at a temperature between 10°C and 20°C.

19. A process as claimed in claim 1 , characterised by effecting ozonization of terpene or of a derivative thereof pure or contained in high concentration in a natural substance, by contact between it, dissolved in an oleic acid ester, and a gaseous mixture containing ozone at a temperature between 10°C and 20°C.

20. Use of the drug claimed in claim 1 , for treating rheumatic affections by external application.

21. Use of the drug claimed in claim 1 , for treating arthrosis by external application.

22. Use of the drug claimed in claim 1 , as a pain-reliever by external application.

23. Use of the drug claimed in claim 1, for treating keratitis.

24. Use of the drug claimed in claim 1 , for treating keloid scars.

25. Use of the drug claimed in claim 1, for treating hematomas.

26. Use of the drug claimed in claim 1 , for treating epidermophytosis.

27. Use of the drug claimed in claim 1 , for treating cutaneous hyperpigmentation phenomena.

28. Use of the drug claimed in claim 1 , for treating wounds.

29. Use of the drug claimed in claim 1 , for treating necrotic wounds.

30. Use of the drug claimed in claim 1, for treating cutaneous fistulas.

31. Use of the drug claimed in claim 1 , for treating chilblains.

32. Use of the drug claimed in claim 1 , for treating Herpes genitalis.

33. Use of the drug claimed in claim 1 , for treating Herpes simplex.

34. Use of the drug claimed in claim 1, for treating skin impurities.

35. Use of the drug claimed in claim 1 , for treating infections of the dental root canal.

36. Use of the drug claimed in claim 1 , for treating dental hyperesthesia.

37. Use of the drug claimed in claim 1 , for treating indurative hypodermitis.

38. Use of the drug claimed in claim 1 as a nematocide.

39. Use of the drug claimed in claim 1 , for treating intestinal infections.

40. A process as claimed in claim 1 , for applying the product to normalize intestinal functions.

41. Use of the drug claimed in claim 1, for treating onychomycosis.

42. Use of the drug claimed in claim 1 , for treating otitis.

43. Use of the drug claimed in claim 1, for treating bed-sores.

44. Use of the drug claimed in claim 1 , for treating athlete's foot.

45. Use of the drug claimed in claim 1, for treating teat infections in cows.

46. Use of the drug claimed in claim 1 , for treating rhinitis of various types.

47. Use of the drug claimed in claim 1 , for treating skin stretch marks.

48. Use of the drug claimed in claim 1 , for treating surgical wounds.

49. Use of the drug claimed in claim 1 , for treating warts.

50. Use of the drug claimed in claim 1 , for externally treating reduced venous-lymphatic circulation.

51. Use of the drug claimed in claim 1, for externally treating infections by mites.

52. Use of the drug claimed in claim 1 , for orally treating the gastroduodenal ulcer.

53. Use of the drug claimed in claim 1 , for treating ulcerations of the lower limbs.

54. Use of the drug claimed in claim 1 , as an external vasodilator.

55. Use of the drug claimed in claim 1 , for treatment as a personal deodorant.

56. Use of the drug claimed in claim 1 , for treating the affections of claims 20, 21, 22, 25, 28, 29, 30, 35, 38, 39, 48 and 51 in either a human or veterinary environment.