WO2008056389A1

WO2008056389A1 - Drug based on free ozonized oleic acid, process for its preparation, and use of the drug

Info

Publication number: WO2008056389A1
Application number: PCT/IT2007/000766
Authority: WO
Inventors: Flavio Torresin; Gualtiero Valeri; Ruggero Manni
Original assignee: Istituto Fitofarmaceutico Euganeo S.R.L.
Priority date: 2006-11-06
Filing date: 2007-10-31
Publication date: 2008-05-15
Also published as: ITFE20060031A1

Abstract

A drug, characterised by consisting of the ozonide of free oleic acid.

Description

DRUG BASED ON FREE OZONIZED OLEIC ACID, PROCESS FOR ITS PREPARATION, AND USE OF THE DRUG

The present invention relates to a drug based on ozonized free oleic acid, a process for its preparation, 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) - which can then open by fragmenting the original chain, to generate new aldehyde and carboxylic groups.

Ozone is particularly soluble in fats, hence even for example free glyceryl stearate could retain small dissolved quantities of free ozone, but this would be unstable or only minimally stable because of the formation of organic ozone derivatives.

As is known from organic analytical chemistry, 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, the following two 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 (eg. 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, hence the ability to reach more or less deep regions by diffusion (the lower the molecular weight, the higher the diffusion rate. Determination of the density change of oils by ozonization appeared for the first time in US 2,083,572, and has been further independently studied in depth, to provide data which is of considerable importance because on an industrial scale it gives an easily measured absolute reference for the degree of ozonization achieved, which has to be corrected only for temperature, this varying during the process. A sharp density increase is always observed, probably determined by the quantity of ozone added t o the double bond and the formation of oxygen bridges between the molecular chains (an effect however which it is sought to avoid during preparation by controlling the temperature, given that the reaction gives rise to pharmacologically inactive compounds which increase the product viscosity). It has been determined that the density increase is virtually in linear relationship with the increase in the ozonide concentration in the oil.

By LC-MS analysis, the compounds formed by the action of ozone on anhydrous oleic acid were found to be fragmented, indicated that with all probability the substance mainly formed is the ozonide of oleic acid which, under normal atmospheric conditions in an anhydrous environment without reducing agents, is reasonably stable and survives for long periods. It has been noted that the product maintains its pharmacological activity unchanged even after many months from preparation, even if preserved at ambient temperature without any regard for exposure to light. By extension, it can be assumed that ozonides are also formed by the action of ozone on other anhydrous unsaturated fats, and that these ozonides are able to behave as liposoluble oxidants and develop free oxygen.

It can be also 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. Ozonization processes have also been proposed for oils such as free coconut oil, free castor oil and triglycerides of ricinoleic acid, olive oil and triglycerides of oleic acid, sesame oil, peanut oil, etc., to obtain drugs, however all these processes have produced drugs of limited reliability and range of use, given the high molecular weight of the derived compounds and the reduced formation of ozonides, or the formation of ozonides of poor activity and unstable with time, or the formation of by-products - epoxides and oxygen bridges between the chains - which are organoleptically unpleasant, pharmacologically inactive or even toxic and irritant. According to the invention these drawbacks are eliminated by a drug 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 anhydrous free oleic acid 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.

Specifically, the process consists of bringing a stream of ozonized gas (for example an oxygen/ozone or air/ozone mixture) into contact with anhydrous free oleic acid or in mixture with other fats, paraffin or apolar solvents, using a suitable contact device (for example, a bubbler of Dreschel bottle type, a plate column or a packed column) and maintaining the temperature around 20°C (not greater than 30⁰C)₁ as the reaction is exothermic. In this respect, at higher temperature the ozone decomposes to form nascent oxygen, which favours the formation of oxygen bridges between the molecular chains (viscous compounds without pharmacological activity, which would degrade the organoleptic characteristics of the product) and epoxides (potentially toxic and irritant). High temperatures also favour decomposition of the oleic acid ozonides. In contrast, if operating at lower temperatures, the oleic acid viscosity increases, and contact between the gas and liquid becomes less efficient. Moreover, below 14-16°C, the oleic acid solidifies, making the reaction impossible. The degree of ozonization is determined by monitoring density, as this increases with the degree of ozonization, the reaction being interrupted when the required density is reached.

The product obtained can be used as such, or formulated in the form of ointments, pomades, oils or gels, which must always exclude water from their composition. The formulation must also avoid reducing or acid reaction substances, which would destroy the contained ozonides. The two most important merits of the drug based on ozonized oleic acid are the following:

- a molecular weight generally equal to about one third (282.45 daltons, against the more than 900 daltons on an average for a fat) giving greater ease of diffusion through tissues, as already stated; - a higher specific active oxygen content, i.e. a greater ratio of active oxygen/moles of substance.

Moreover, by treating anhydrous free oleic acid, it remains stable because of the impossibility of the formed ozonides becoming hydrolysed by the action of the water. It has been verified experimentally that the pharmacological activity of ozonized oleic acid is considerably higher than that of other ozonized unsaturated fats; its organoleptic characteristics are also decidedly better, especially with regard to viscosity, which for ozonized oleic acid is much less than that of other ozonized oils.

With regard to viscosity, oleic acid does not increase its viscosity much on ozonization, this favouring the process; in contrast, in a case such as that of castor oil, the viscosity increases to a point such that it is impossible to pursue ozonization. A consistent viscosity increase is also an indication of polymer formation; we have already seen that a molecular weight increase is a negative circumstance for the pharmacological activity of the product. The action of ozone on the oleic acid 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.

It has been found possible to improve the pharmacological effects of the product obtained by mixing with it, in suitable manner, medicinal plant extracts, essential oils or other active principles which cannot be oxidized by the ozonized oleic acid or decompose it. Particularly effective against bedsores has proved to be ozonized oleic acid in association with α-lipoic acid, which exalts its natural vulnerary power. On the basis of theoretical and practical studies, ozonized oleic acid presents 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

Various other uses of cosmetic type are possible, in particular for cleaning and disinfecting the skin.

Essentially, ozonized oleic acid has proved to possess pharmacological activity similar to that already found for ozonized oils, but much more rapid and intense, in addition to showing considerable effectiveness in various affections for which possible treatment with ozonized oils has never before been observed or described.

The property of ozonized organic compounds to block virus propagation through tissues, as in the case of ozone, is considerable. By way of non-limiting example, the following represent some of the more interesting cases of ozonized oleic acid produced in the aforesaid manner and prepared in oils, ointments, pomades or oleogels, or enclosed in capsules or pearls for internal use, free 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.

EXAMPLE 1

This case relates to a patient aged 47 years suffering from a form of psoriasis documented for five years. The patient was under treatment with cortisone. The psoriasis manifested as widely distributed blisters on the hands and feet. The form on the feet often determined difficulties such as to compel the patient to take to bed. The treatment with cortisone gave no substantial improvement, beyond checking the spread.

The patient was treated with two applications of ozonized oleic acid per day for four weeks starting from October 2005. After two weeks of treatment, the patient presented a substantial reduction in the phenomenon, which on the hands was reduced to merely a slight reddening.

After four weeks the phenomenon had virtually disappeared.

The patient decided to suspend cortisone treatment and continue treatment only with ozonized oleic acid.

EXAMPLE 2

This case in question related to a patient aged 47 years suffering for two years from a form of psoriasis, according to diagnosis. The patient had attempted treatment with various preparations without substantial improvement. The psoriasis manifested as wide desquamation from the hands to the elbows, where manifestations of erythematous squamous type were present.

The patient was treated with two applications of ozonized oleic acid per day for three weeks starting from June 2005. After two weeks of treatment, the patient presented almost complete regression. The patient decided to continue treatment only with ozonized oleic acid.

EXAMPLE 3

This case in question related to a patient aged 51 years suffering from psoriasis (erythematous squamous stains) at the elbows and face, with extension to the lower limbs, under treatment for two years. After about forty days of treatment with ozonized oleic acid, the stains disappeared.

EXAMPLE 4

Female patient aged 56 years suffering from erythematous stains similar to psoriasis at the thighs and abdomen for a period of six months.

After about 25 days of treatment with ozonized oleic acid, the stains disappeared.

EXAMPLE 5

Female patient aged 65 years with arthrous and varicose pains in the lower limbs and blue-coloured venous stasis at the ankles, persisting for many years.

The patient was treated with ozonized oleic acid twice a week.

Regression of the varicose veins and venous stasis was noted, with lesser heaviness in the legs. EXAMPLE 6

Female patient aged 35 years with arthrous and varicose pains in the lower limbs and blue-coloured venous stasis at the ankles, persisting for many years.

After about six weeks of treatment with ozonized oleic acid the manifestations had almost completely regressed.

EXAMPLE 7

Male patient aged 82 years with varicose ulcer in one leg at the level of the malleolus, persisting for almost a year; after five months of treatment with ozonized oleic acid the varicose ulcer disappeared. EXAMPLE 8 Female patient aged 45 years suffering from external haemorrhoids for about three and a half years.

After 15 days of ozonized oleic acid applications the external haemorrhoids disappeared. EXAMPLE 9

Male patient aged 45 years suffering from eczema on the sole of the feet.

The patient applied ozonized oleic acid twice a day, morning and evening, on the affected region. After ten days of treatment the symptoms had virtually disappeared.

EXAMPLE 10

Male patient aged 45 years with widespread periodic skin irritation in the region between the groin and the thighs. The phenomenon appears suddenly at irregular intervals. Application of ozonized oleic acid determined its complete and stable regression within two hours.

EXAMPLE 11

Male patient aged 48 years with a form of eczema on the chin.

The patient was treated with two applications of ozonized oleic acid per day.

After a week of treatment, the eczematous form regressed completely.

EXAMPLE 12

Female patient aged 68 years with wound on the back of the left hand.

Application of ozonized oleic acid determines accelerated cicatrization of the wound without further consequences.

EXAMPLE 13 Female patient aged 68 years with slight burn on the right arm, determined by kitchen accident (hot oil).

Application of ozonized oleic acid determines accelerated healing of the burn without further consequences. EXAMPLE 14

Male patient aged 31 years, suffering from cephalic crises concentrated at the weekends.

The patient was under phytotherapeutic treatment, followed a suitable dietary regime, cyclic massage periods. With this, the cephalic crises were reduced from an average of three per week to one. The patient also made habitual use of synthetic antiinflammatories to mitigate the pain.

With the commencement of regular application of ozonized oleic acid to the base of the neck once a day for one month, the painful manifestations disappear, to reappear when the treatment is suspended.

EXAMPLE 15

Female patient aged 68 years with widespread articular pains.

Application of ozonized oleic acid proves able to calm the pain, starting from the initial application. EXAMPLE 16

Male patient aged 62 years, with duodenal ulcer and Barrett's esophagus.

When subjected to gastroscopy, the patient shows serious widespread ulceration of the gastric mucosa, which in the previous days had also given rise to profuse haemorrhage. Ozonized oleic acid in 0.75 g capsules was administered twice a day after main meals, for a period of two weeks.

After the two weeks the patient was subjected to further gastroscopy, showing complete regression of the gastric mucosa ulceration. EXAMPLE 17

Male patient aged 53 years, with purulent perianal fistula persisting for two years.

The patient was treated for three months with ozonized oleic acid.

The fistula dried and gave no further purulent eliminations. EXAMPLE 18

Male patient aged 45 years, with external haemorrhoids persisting for a few days.

The patient was treated for three days with ozonized oleic acid.

The haemorrhoids completely regressed. EXAMPLE 19

Male patient aged 46 years, with consequences of a surgical operation at the knee found under arthroscopy. Fifteen days after the operation the surgical wounds, although healed, were still very sensitive to pressure and the knee appeared rather inflamed. After two days of applying the preparation twice a day, the swelling had disappeared and the surgical wounds had lost the painfulness initially complained of by the patient.

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

Claims

C L A I M S

1. A drug characterised by consisting of the ozonide of free oleic acid.

2. A process for preparing the drug claimed in claim 1 , characterised by subjecting free anhydrous oleic acid to ozonization at a temperature between 20 and 30°C.

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

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

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

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

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

8. A process as claimed in claim 2, characterised by effecting ozonization with an ozone/inert gas mixture.

9. A process as claimed in claim 2, characterised by effecting ozonization with a mixture of ozone/oxygen/nitrogen in various proportions.

10. A process as claimed in claim 2, characterised by mixing the oleic acid with vegetable oils.

11. A process as claimed in claim 2, characterised by mixing the oleic acid with animal oils.

12. A process as claimed in claim 2, characterised by mixing the oleic acid with apolar solvents.

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

14. Use of the drug claimed in claim 1 , for externally treating psoriasis.

15. Use of the drug claimed in claim 1 , for externally treating eczema.

16. Use of the drug claimed in claim 1 , for treating burns.

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

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

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

20. Use of the drug claimed in claim 1 , for treating vaginitis and vulvovaginitis.

21. Use of the drug claimed in claim 1 , for treating rheumatic affections.

22. Use of the drug claimed in claim 1 , as a general bactericide.

23. Use of the drug claimed in claim 1, as a pain-reliever for external use.

24. Use of the drug claimed in claim 1 as a general antimycotic for external use.

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

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

27. Use of the drug claimed in claim 1 , for treating Candida infections.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

44. Use of the drug claimed in claim 1 , for treating intestinal protozoa.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

59. Use of the drug claimed in claim 1, for externally treating infections by mites. - I8 ¬

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

61. Use of the drug claimed in claim 1 , as a vasodilator for external use.

62. A drug claimed in claim 1 , in the form of a preparation, free liquid or in mixture with other active principles.

63. A drug claimed in claim 1 , in the form of a compound in oils, ointments or pomades, or oleogels.

64. A drug claimed in claim 1, enclosed in capsules or pearls for internal use, either free or mixed with excipients.

65. A drug claimed in claim 1 , as a microencapsulated preparation or reduced to ovule form or spray form.

66. Use of the drug claimed in claim 1 , for treating the affections of claims 16, 17, 18, 19, 20, 21 , 22, 23, 24, 26, 30, 37, 43, 44, 47, 48, 49, 51 , 57 and 59 in either a human or veterinary environment.