WO2017111569A1 - Pharmaceutical combination and formulation for the treatment of eye diseases - Google Patents

Abstract

The invention relates to a pharmaceutical combination based on serratiopeptidase as an anti-inflammatory and analgesic, at least one quinolone as an antimicrobial, and an amino acid as a stabiliser of the serratiopeptidase. The invention also relates to a pharmaceutical formulation for the treatment of eye diseases, comprising said pharmaceutical combination, a thickening agent, an excipient and double-distilled water. The invention further relates to a method for preparing said formulation, comprising: producing a first mixture of serratiopeptidase, at least one quinolone, an amino acid, at least one liposome and double-distilled water, mixing with an agitation of 500-700 rpm; producing a second mixture of the thickener with double-distilled water, mixing with an agitation of 500 rpm for 20 mins; sterilising the mixture with wet steam at 120°C for 15 mins; and cooling same to approximately 30-36°C; and producing a third mixture, comprising the mixing of the excipients of said formulation and mixing the first, second and third mixture.

Description

 COMBINATION AND PHARMACEUTICAL FORMULATION, FOR THE

 TREATMENT OF EYE DISEASES

TECHNICAL FIELD OF THE INVENTION

The present invention relates to the technical field in Chemistry and Pharmaceuticals, since it provides a combination and pharmaceutical formulation for the treatment of eye diseases, such as inflammation, microbial infection, pain, and combinations among them; To name a few of them.

BACKGROUND OF THE INVENTION Currently, it has been found that eye diseases are caused by several factors or agents, for example microbial agents, inflammatory agents, pain-causing agents, among others. That is why the need to create combinations of active ingredients that act together synergistically, for the proper control of such diseases. In this regard, research has already been carried out and products or mixtures of active substances have been created to treat eye disease cocktails. Among the drawbacks of such current products or mixtures, is that they do not have a lasting pharmacological stability, especially in those products or mixtures that include biological and / or genetic material. Singh et al. (2014) disclosed a study whose objective was to explore the development of a periodontal system of dual-controlled release, of a fluoroquinolone (ciprofloxacin) and an anti-inflammatory (serratiopeptidase enzyme). Both active ingredients were applied in a dose of 30 mg of ciprofloxacin hydrochloride and 10 mg of serratiopeptidase. Pluronic F127 (900 to 1100 mg), carbopol (25 to 75 mg) were also added. Stability studies were measured, where the formulation: 30 mg of ciprofloxacin hydrochloride, 10 mg of serratiopeptidase, 1100 mg of pluronic F127, and 75 mg of carbopol, was stable when stored at 25 ° C, at 60% relative humidity , and at 40 ° C, 75% humidity, over a period of 90 days. Although the use of a mixture of ciprofloxacin and serratiopeptidase is already proposed, to develop a system of release and stability of the mixture, but it is not indicated that the mixture could be used to treat eye diseases.

For their part, Selan et al. (1993) report that among the different mechanisms of bacterial resistance to the antimicrobials that have been studied, the formation of biofilms is one of the most widespread. This mechanism is often the cause of failure in the treatment of infections of prosthetic devices, and several attempts have been made to develop molecules and protocols that are capable of inhibiting biofilm-embedded bacteria. Data are presented that suggest the possibility that proteolytic enzymes could significantly improve the activities of antibiotics against biofilm (biofilm). Antibiotic sensitivity tests in both planktonic and sessile cultures, studies on the dynamics of colonization of 10 isolates of biofilm formation, and then bioluminescence and scanning electron microscopy under seven different experimental conditions showed that Serratiopeptidase greatly improves the activity of ofloxacin in sessile cultures and can inhibit the formation of biofilms. As can be seen, in this document, the use of the mixture of Serratiopeptidase with Ofloxacin is already being disclosed, with medicinal applications and / or uses for the prevention of biofilms or infections caused by bacteria in humans, with prostheses; however nothing is described about the stabilization of Serratiopeptidase.

Patent document MX2013013286 (A) describes a formulation and method of preparation for obtaining ophthalmic solutions derived from quinolones and a Serratiopeptidase enzyme with anti-inflammatory and analgesic properties. Where they were able to obtain pharmacologically stable formulations, even when the active ingredients are of a different nature, since they were able to conserve their biological activity and efficiency during the storage and handling time. Stability was evaluated at 60 and 120 days for the accelerated condition; and at 60, 120, 180 and 360 days for the long term condition. However, such stability is not sufficient because the exposure and absorption time of the active ingredients in the eyeball is greatly reduced and the half-life and effect of the serratiopeptidase enzyme is greatly reduced.

Due to the aforementioned drawbacks, a pharmaceutical combination and formulation was developed to treat eye diseases with lasting stability for the shelf life of the pharmaceutical formulation. A method was also developed to prepare said formulation. DETAILED DESCRIPTION OF THE INVENTION

The characteristic details of the present invention are clearly shown in the following description and accompanying examples, which illustrate some of the preferred embodiments of the present invention, whereby said description and examples are merely illustrative, but not limiting to said invention. In the first instance, the present invention relates to a pharmaceutical combination of: i) Serratiopeptidase, as an anti-inflammatory and analgesic agent;

ii) at least one Quinolone, as an antimicrobial agent; and iii) an amino acid, as a stabilizing agent for Serratiopeptidase.

Serratiopeptidase can be of any origin, as long as it has the function of acting as an anti-inflammatory and analgesic agent. One modality is for said Serratiopeptides to be obtained from the E-15 strain of Serratia Sp. As for Quinolones, these can be of any generation, with those of broad antimicrobial spectrum being preferred, for example: Ciprofloxacin, Moxifloxanine, Gatifloxacin, su combination between them, to name a few.

A preferred embodiment of the invention is that Quinolone is Ciprofloxacin. In relation to amino acids, those that provide stability to Serratiopeptidase, such as, L-Arginine and Lysine, are preferred. The amino acid L-Arginine is still more preferred. The fact of including an amino acid to the pharmaceutical combination, a stability of up to 2 years is achieved, a technical advantage that the known formulations do not have.

The combination obtained in the present invention can be used for the preparation of pharmaceutical formulations, medicaments, pharmaceutical solutions, etc., to treat diseases, preferably diseases that affect the ocular areas of an animal, including the human.

Therefore, in this case, the present invention relates to a pharmaceutical formulation for the treatment of eye diseases, which comprises a pharmaceutical combination in accordance with the present invention; a pharmaceutically acceptable viscosifying agent; at least one pharmaceutically acceptable excipient; and double distilled water.

Where the components of the pharmaceutical combination are in the following proportions: Serratiopeptidase from 0.22 to 0.35%; quinolone from 0.28 to 030%; and the amino acid from 0.05 to 0.09%; with respect to the total weight of the pharmaceutical formulation. Where the viscosifying agent useful for the pharmaceutical formulation of the present invention may be carboxymethyl cellulose, which may be in a concentration of 0.05 to 0.13%, with respect to the total weight of the pharmaceutical formulation.

Within the pharmaceutically acceptable excipients that can be used in the pharmaceutical formulation, in question, it is a mixture that can comprise: Liposomes, Polyoxyl Stearate, Sodium Dibasic Phosphate, Sodium Monobasic Phosphate, and EDTA. A preferred embodiment of the pharmaceutical formulation of the present invention is when the excipients are in the following amounts: the Liposomes are in an amount of 0.92 to 1.02%, the Polyoxyl Stearate is 0.11 to 0.18%, Sodium Dibasic Phosphate in 0.16 to 0.22%, Sodium Monobasic Phosphate in 0.27 to 0.31%, and EDTA in 0.05 to 0.13%.

A further embodiment of the present invention is when the pharmaceutical formulation may optionally comprise Benzalkonium Chloride, which may be in an amount of 0.001 to 4%; As an antimicrobial agent.

The pharmaceutical formulation in question includes a diluent to dissolve the ingredients, for example the most common diluent is double-distilled water, whose required amount is sufficient to dissolve and mix the ingredients of the formulation, in this case, double-distilled water it is in a proportion of 97.25 to 97.77%, with respect to the total weight of the pharmaceutical formulation.

However, the diluent can be substituted by other substances, according to the presentation that you want to give to the pharmaceutical formulation in question; by For example, presentations in ointments, ointments, creams, spray, among others.

It is very important to mention that the pharmaceutical formulation must have a pH of 5.5 to 7.5, so that there are no undesirable phenomena, such as irritation, discomfort to instillation, etc.

Said formulation must have an osmolarity similar to that of a tear, to avoid phenomena of low absorption or local irritation. Therefore, it is suggested that said osmolarity be around 300 Mosm / L.

The eye disease that can treat the formulation of the present invention can be: an inflammation, microbial infection, pain, and combinations among them; To name a few of them.

The present invention also relates to a method for preparing a pharmaceutical formulation, for the treatment of eye diseases, such as the pharmaceutical formulation described in the present invention; Said method comprises the following steps: i) Obtain a first mixture, which in turn comprises mixing the Serratiopeptidase in an amount of 0.22 to 0.35%, at least one Quinolone in an amount of 0.28 to 0.30%, an Amino Acid in a amount of 0.05 to 0.09%, at least one Liposome in a proportion of 0.92 to 1.02%, and double distilled water in an amount of 40% with respect to the total weight of the pharmaceutical formulation; where the mixing is with a stirring of 500 to 700 rpm. Obtain a second mixture, which in turn comprises dissolving a pharmaceutical grade viscosifying agent, in an amount of 0.05 to 0.13%, with double-distilled water in an amount of 20% with respect to the total volume of said mixture; the mixture was made with stirring at 500 rpm, for 20 min; sterilize the mixture with wet steam at 120 ° C, for 15 min; and cool the mixture to room temperature, at about 30 to 36 ° C. The viscosifying agent may be pharmaceutically acceptable carboxymethyl cellulose. iii) Obtain a third mixture, comprising mixing the excipients: Polyoxyl Stearate present in an amount of 0.11 to 0.18%, in double distilled water in a concentration of 25% of the total weight of the pharmaceutical formulation; heat the mixture to 70 ° C, with stirring of 500 rpm; cool the mixture to room temperature; and immediately added: Sodium Dibasic Phosphate in an amount of 0.16 to 0.22%, Monobasic Sodium Phosphate in a proportion of 0.27 to 0.31%, and EDTA in an amount of 0.05 to 0.13%. iv) Stir the third mixture constantly 500 rpm.

Add the second mixture to the third mixture, by constant stirring of 500 rpm, for 35 min under stirring. vi) Then add the first mixture to the previous mixture with the same stirring for 35 min; vii) Aforate the mixture obtained with double-distilled water, adding the rest of the water volume, which is about 15%, with constant stirring of 1500 rpm, for 35 min. viii) Finally, the pH of the formulation is adjusted from 5.5 to 7.5, and an osmolarity of 300 Mosm / L.

One embodiment of the method of the present invention is that Serratiopeptidase is originated from strain E-15 of Serratia Sp.

A further embodiment of the present method is when quinolone is selected from the following group: Ciprofloxacin, Moxifloxanino, Gatifloxacin, and their combination among them. Being more preferred when quinolone is Ciprofloxacin.

A preferred embodiment of the method in question is when the amino acid is L-Arginine or Lysine; even more so when said amino acid is L-Arginine.

Another embodiment of said method is that it can be added to the pharmaceutical formulation, optionally, Benzalkonium Chloride; which can be in an amount of 0.001 to 4%.

Therefore, the present invention also relates to the use of a pharmaceutical combination comprising: Serratiopeptidase, at least one Quinolone, and one amino acid; for the preparation of a pharmaceutical formulation useful in the treatment of eye diseases in a patient suffering from them. A variant of the use of the pharmaceutical combination of the present invention is when the application of the pharmaceutical formulation, to the patient suffering from eye diseases, is that on the first day, a drop of the formulation is applied for each affected eye, every 30 min, during the first 6 h, then one drop per eye every hour for the rest of the day; on the second day, one drop per eye every 2 h; and from the third to the tenth day, one drop per eye every 4 h.

A further modality of the use of the pharmaceutical combination, in question is when the eye disease is: an inflammation, microbial infection, pain, and combinations between them.

Ejaoploa

Example 1. This example illustrates one embodiment of the pharmaceutical formulation for the treatment of eye diseases, of the present invention, which is merely illustrative, and therefore should not be considered as a limitation for the present invention. Table 2. Components of the pharmaceutical formulation for the treatment of eye diseases, of the present invention.

1

one

Example 3. This example illustrates a method of preparing a pharmaceutical formulation for the treatment of eye diseases _/ which is merely a preferred embodiment of the invention _/ so it should not be considered as a limitation for said invention.

For a better or optimal preparation of the formulation, it is first preferred to prepare the following mixtures separately.

Mixture 1, which corresponds to the mixture of active ingredients and natural liposomes.

3 mg of Serratiopeptidase obtained from strain E-15 of Serratia Sp. 3 mg of Ciprofloxacin were mixed; 0.75 mg of L-Arginine; and 10 mg of liposomes; with a stirring of 500 to 700 rpm. Then 0.4 mL of double-distilled water was added.

Mixture 2, for the dissolution of a viscosifying agent.

1 mg of pharmaceutical grade carboxymethylcellulose was dissolved in 0.2 mL of double distilled water, with stirring of 500 rpm, for 20 min. The mixture was sterilized with wet steam at 120 ° C for 15 min. It was then allowed to cool to room temperature, around 33 ° C. Mixture 3, the rest of the necessary excipients are dissolved in this mixture.

1.5 mg of Polyoxyl Stearate was dissolved in 0.25 mL of double distilled water, heating the mixture at 70 ° C, with stirring of 500 rpm. The mixture is then allowed to cool to room temperature; and immediately added: 2 mg of Sodium Dibasic Phosphate, 3 mg of Sodium Monobasic Phosphate, and 1 mg of EDTA.

Once the mixtures were prepared, they were stirred as follows:

Ά mixture 3 was subjected to constant stirring of 1500 rpm, and mixture 2 (viscous) was added and allowed to stir for 35 min. Then add mixture 1 with the same stirring for an additional 35 min. Subsequently, it was fed with double-distilled water by adding a volume of 0.15 mL, with constant stirring of 1500 rpm, for 35 min, which achieves a complete integration of the ingredients involved in the present formulation.

Finally, the pH obtained was adjusted to pH 6.5, with HC1, and an osmolarity of 300 Mosm / L.

Claims

CLAIMS 1. A pharmaceutical combination, characterized in that it comprises: i) Serratiopeptidase, as an anti-inflammatory and analgesic agent;  ii) at least one Quinolone, as an antimicrobial agent; Y  iii) an amino acid, as a stabilizing agent for Serratiopeptidase. 2. The combination of the preceding claim, wherein Serratiopeptidase is produced by strain E-15 of Serratia Sp. 3. The combination according to claim 1, wherein the Quinolone is selected from the following group: Ciprofloxacin, Moxifloxanino, Gatifloxacin, and their combination among them. 4. The combination of the preceding claim, wherein the Quinolone is Ciprofloxacin. 5. The combination according to claim 1, wherein the amino acid is L-Arginine or Lysine. 6. The combination according to the preceding claim, wherein the amino acid is L-Arginine. 7. A pharmaceutical formulation for the treatment of eye diseases, characterized in that it comprises: i) a pharmaceutical combination which in turn comprises:  to. Serratiopeptidase, as an anti-inflammatory and analgesic agent;  b. at least one Quinolone, as an antimicrobial agent; Y  C. an amino acid, as a stabilizing agent for Serratiopeptidase;  ii) a pharmaceutically acceptable viscosifying agent; iii) a pharmaceutically acceptable excipient; and iv) double distilled water. 8. The formulation of the preceding claim, wherein the Serratiopeptidase is produced by the strain E-15 of Serratia Sp., And is present in an amount of 0.22 to 0.35%, relative to the total weight of the formulation. 9. The formulation according to claim 7, wherein the Quinolone is selected from the following group: Ciprofloxacin, Moxifloxanino, Gatifloxacin, and their combination among them. 10. The formulation of the preceding claim, wherein the quinolone is Ciprofloxacin and is present in an amount of 0.28 to 0.30%, based on the total weight of said pharmaceutical formulation. 11. The formulation according to claim 7, wherein the amino acid is L-Arginine or Lysine. 12. The formulation according to the preceding claim, wherein the amino acid is L-Arginine and is present in a proportion of 0.05 to 0.09%, in relation to the total weight of the pharmaceutical formulation. 13. The formulation of the preceding claim, wherein the viscosifying agent is carboxymethyl cellulose and is in a concentration of 0.05 to 0. 13%, with respect to the total weight of such formulation. 14. The formulation according to claim 7, wherein the pharmaceutically acceptable excipient is a mixture comprising: Liposomes, Polyoxyl Stearate, Dibasic Sodium Phosphate, Sodium Monobasic Phosphate, and EDTA. 15. The formulation according to claim 9, wherein the Liposomes are in an amount of 0.92 to 1.02%, the Polyoxyl Stearate 0.11 to 0.18%, Sodium Dibasic Phosphate from 0.16 to 0.22%, Sodium Monobasic Phosphate from 0.27 to 0.31 %, and EDTA from 0.05 to 0.13%; in relation to the total weight of the pharmaceutical formulation. 16. The formulation as claimed in claim 7, wherein the double distilled water is in a proportion of 97.25 to 97.77%, based on the total weight of the formulation. 17. The formulation according to claim 7, wherein said formulation has a pH of 5.5 to 7.5; and an osmolarity of 300 Mosm / L. 18. The formulation of claim 7, characterized in that it optionally comprises Benzalkonium Chloride. 19. The formulation according to the preceding claim, wherein the Benzalkonium Chloride is in an amount of 0.001 to 4%, based on the total weight of said pharmaceutical formulation. 20. The formulation according to claim 7, wherein the eye disease is: inflammation, microbial infection, pain, and combinations between them. 21. A method for preparing a pharmaceutical formulation, for the treatment of eye diseases, characterized in that it comprises: obtaining a first mixture, which in turn comprises mixing the Serratiopeptidase in an amount of 0.22 to 0.35%, at least one Quinolone in an amount of 0.28 to 0.30%, an Amino acid in an amount of 0.05 to 0.09%, at least one Liposome in a proportion of 0.92 to 1.02%, and bidistilled water in an amount of 40%, with respect to the total weight of the pharmaceutical formulation; where mixing is with stirring of 500 to 700 rpm; obtaining a second mixture, which in turn comprises dissolving a pharmaceutical grade viscosifying agent, in an amount of 0.05 to 0.13%, with double-distilled water in an amount of 20%, based on the total weight of said mixture; the mixture was made with stirring at 500 rpm, for 20 min; sterilize the mixture with wet steam at 120 ° C, for 15 min; and cool the mixture at room temperature, at about 30 to 36 ° C;  iii) obtaining a third mixture, comprising mixing the excipients: Polyoxyl Stearate present in an amount of 0.11 to 0.18%, in double distilled water in a concentration of 25% in relation to the total weight of the pharmaceutical formulation; heat the mixture to 70 ° C, with stirring of 500 rpm; cool the mixture to room temperature; and immediately added: Dibasic Sodium Phosphate in an amount of 0.16 to 0.22%, Monobasic Sodium Phosphate in a proportion of 0.27 to 0.31%, and EDTA in an amount of 0.05 to 0.13%, in relation to the total weight of said formulation;  iv) stir the third mixture constantly at 500 rpm;  v) add the second mixture to the third mixture, by constant stirring of 500 rpm, for 35 min under stirring;  vi) add the first mixture to the previous mixture with the same stirring for 35 min; vii) fill the obtained mixture with double-distilled water, adding the rest of the water volume, which is about 15%, with constant stirring of 1500 rpm, for 35 min; Y  viii) adjust the pH of the formulation from 5.5 to 7.5, and an osmolarity of 300 Mosm / L. 22. The method of the preceding claim, wherein Serratiopeptidase is from strain E-15 of Serratia Sp. 23. The method according to claim 21, wherein the quinolone is selected from the following group: Ciprofloxacin, Moxifloxanino, Gatifloxacin, and their combination among them. 24. The method of the preceding claim, wherein the quinolone is Ciprofloxacin. 25. The method according to claim 21, wherein the amino acid is L-Arginine or Lysine. 26. The method according to the preceding claim, wherein the amino acid is L-Arginine. The method of claim 21, wherein the viscosifying agent is carboxymethyl cellulose. The method of the preceding claim, characterized in that it comprises optionally adding, to the pharmaceutical formulation, Benzalkonium Chloride. 29. The method according to claim 21, wherein the Benzalkonium Chloride is in an amount of 0.001 to 4%. 30. The use of a combination comprising:  Serratiopeptidase, at least one Quinolone, and one amino acid; for the preparation of a pharmaceutical formulation to treat eye diseases. 31. The use according to the preceding claim, wherein the first day application is a drop of the pharmaceutical formulation for each affected eye, every 30 min, during the first 6 hours, then one drop per eye every hour for the rest of the day; on the second day, one drop per eye every 2 h; and from the third to the tenth day, one drop per eye every 4 h. 32. The use according to claim 26, wherein the eye disease is: inflammation, microbial infection, pain, and combinations between them.